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Issue 13 ， 2022
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Protective Effect of Linggui Zhugantang Medicated Serum Against H2O2-induced Injury in H9c2 Cells by Regulating PI3K/Akt Signaling Pathway
Abstract：ObjectiveTo investigate the protective effect of Linggui Zhugantang （LGZGT）-medicated serum against H2O2-induced injury in H9c2 cells and its relationship with the phosphatidylinositol 3- kinase/protein kinase B （PI3K/Akt） signaling pathway.MethodThe LGZGT-medicated serum and blank serum were prepared based on serum pharmacology. H9c2 cells were cultured in vitro and divided into a normal group， an H2O2 group， a 20% blank serum group， and a 20% LGZGT-medicated serum group. The cells were treated with corresponding drugs for 12 h and cultured with 100 μmol·L-1 H2O2 for another 6 h. The effect of 20% LGZGT-medicated serum on the proliferation activity of H9c2 cells induced by H2O2 was detected by cell counting kit-8 （CCK-8） assay. Mitochondrial reactive oxygen species （ROS） level was detected by the fluorescence probe. The levels of malondialdehyde （MDA）， lactate dehydrogenase （LDH）， catalase （CAT）， and glutathione peroxidase （GSH-Px） were detected by colorimetry. Western blot was used to detect the protein expression levels of phosphoinositide 3-kinase （PI3K）， phosphorylated-PI3K （p-PI3K）， protein kinase B （Akt）， and phosphorylated-Akt （p-Akt）. Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expression of PI3K and Akt. Flow cytometry was used to detect the apoptosis rate. After the addition of PI3K inhibitor LY294002， the levels of mitochondrial ROS， LDH， and GSH-Px， protein expression of PI3K， p-PI3K， Akt， and p-Akt， and cell apoptosis rate were detected.ResultCompared with the normal group， the H2O2 group showed blunted cell viability （P<0.01）， increased levels of mitochondrial ROS， MDA， and LDH （P<0.01）， decreased levels of CAT and GSH-Px （P<0.01）， reduced phosphorylation and mRNA expression of PI3K and Akt （P<0.05， P<0.01）， and increased apoptosis rate （P<0.01）. Compared with the H2O2 group， the 20% LGZGT-medicated serum group showed potentiated cell viability， reduced levels of mitochondrial ROS， MDA， and LDH （P<0.01）， increased levels of CAT and GSH-Px （P<0.01）， up-regulated phosphorylation and mRNA expression of PI3K and Akt （P<0.05， P<0.01）， and decreased apoptosis rate （P<0.01）. The combined use of LGZGT-medicated serum and inhibitor LY294002 reversed the above-mentioned effects of LGZGT-medicated serum on H9c2 cells （P<0.05， P<0.01）.ConclusionThe protective effect of LGZGT-medicated serum on H2O2-induced H9c2 cell injury may be related to the regulation of the PI3K/Akt signaling pathway to reduce oxidative stress and apoptosis.Keywords：Linggui Zhugantang;oxidative stress;apoptosis;phosphatidylinositol 3- kinase/protein kinase B （PI3K/Akt） signaling pathway- 110
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Therapeutic Effect of Buyang Huanwutang on Diabetic Peripheral Neuropathy Rats from Perspective of Oxidative Stress
Abstract：ObjectiveTo explore the neuroprotective mechanism of Buyang Huanwutang （BYHW） on diabetic peripheral neuropathy （DPN） rats based on oxidative stress and investigate the dosage of Astragali Radix （AR）.MethodNinety SD rats were randomly divided into a normal group， a model group， an α-lipoic acid group （60 mg·kg-1·d-1）， and BYHW groups with high- （15 g·kg-1·d-1）， medium- （8.75 g·kg-1·d-1）， and low-dose （5.625 g·kg-1·d-1） AR groups. The diabetes model was induced in rats except for those in the normal group by the high-sugar/high-fat diet and intraperitoneal injection of streptozotocin （STZ）. Drug intervention lasted for 12 weeks. The paw withdrawal threshold （PWT） and sensory nerve conduction velocity （SNCV） were detected after drug intervention. Gonad-stimulating hormone （GSH） and malondialdehyde （MDA） were determined. The mitochondrial morphology and structure in sensory neurons of L4-5 dorsal root ganglion （DRG） of rats were observed by electron microscopy. Respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ activities and the mitochondrial membrane potential were detected. The main proteins in the adenosine monophosphate-activated protein kinase （AMPK）/nuclear factor-related factor-2 （Nrf2） pathway， such as phosphorylated AMPK （p-AMPK）， phosphorylated Nrf2（p-Nrf2）， heme oxygenase-1 （HO-1）， and quinone NADH dehydrogenase 1 （NQO1）， were detected by immunohistochemistry and Western blot.ResultCompared with the normal group， the model group showed increased fasting blood glucose （P<0.01）， decreased content of SNCV， PWT， and GSH （P<0.01）， elevated MDA content （P<0.01）， obvious mitochondrial damage with vacuolations， reduced activities of respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ and mitochondrial membrane potential （P<0.01）， and declining p-AMPK， p-Nrf2， HO-1， and NQO1 （P<0.01）. Compared with the model group， the α-lipoic acid group and BYHW high-dose group showed increased SNCV， PWT， and GSH， decreased MDA （P<0.05， P<0.01）， alleviated mitochondrial structural damage， increased respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ activities and mitochondrial membrane potential （P<0.01）， and elevated p-AMPK， p-Nrf2， HO-1， and NQO1 （P<0.05， P<0.01）.ConclusionBYHW regulates oxidative stress through the AMPK/Nrf2 pathway to treat DPN. The therapeutic effect of BYHW is related to the dosage of AR. The BYHW group with high-dose AR is superior to the BYHW groups with medium- and low-dose AR groups in inhibiting oxidative stress.Keywords：diabetic peripheral neuropathy;Buyang Huanwutang;Astragali Radix;mitochondria;adenosine monophosphate-activated protein kinase （AMPK）/ nuclear factor-related factor-2 （Nrf2） pathway;oxidative stress- 68
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Gegen Qinliantang and Its Combinations Inhibit Oxidative Stress Injury in Ulcerative Colitis Rats by Regulating Nrf2/NQO1 Signaling Pathway
Abstract：ObjectiveTo explore the underlying mechanism of Gegen Qinliantang （GGQL） in the treatment of ulcerative colitis （UC） in rats and discuss the effects of modification of GGQL on its efficacy.MethodThe UC model was induced in rats by free access to 5% dextran sulfate sodium in saline solution. Male SD rats were randomly divided into a normal group， a model group， a positive control group （sulfasalazine enteric-coated tablets， 350 mg·kg-1）， a GGQL group （17 g·kg-1）， a Glycyrrhizae Radix et Rhizoma （GR）-absent GGQL group （17 g·kg-1）， a Puerariae Lobatae Radix （PLR）-absent GGQL group （17 g·kg-1）， a GR-PLR group （17 g·kg-1）， and a Scutellariae Radix （SR）-Coptidis Rhizoma （CR） group （17 g·kg-1）. The in vitro antioxidant activities of GGQL and its combinations were evaluated by 2，2-diphenyl-1-picrylhydrazyl （DPPH）， 2，2′-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS）， and fluorescence recovery after photobleaching （FRAP） methods. The degree of colonic tissue injury in each group was evaluated based on the weight changes of rats， the length of the colon， the colon sections， and hematoxylin-eosin （HE）-stained histopathologic sections. The serum levels of myeloperoxidase （MPO）， lipid peroxide （LPO）， malondialdehyde （MDA）， total superoxide dismutase （T-SOD）， catalase （CAT）， and reduced glutathione （GSH） were measured by colorimetry. The mRNA and protein expression of nuclear factor-erythroid 2 related factor （Nrf2）， quinone oxidoreductase 1 （NQO1）， and heme oxygenase-1 （HO-1） in colon tissues was detected by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.ResultCompared with the normal group， the model group showed colonic mucosal necrosis， inflammatory infiltration， increased serum levels of MPO， LPO， and MDA （P<0.01）， blunted activities of T-SOD， CAT， and GSH （P<0.01）， decreasing trend of mRNA expression of Nrf2， NQO1， and HO-1， reduced expression of Nrf2 protein （P<0.01）， and decreasing trend of expression of NQO1 and HO-1 proteins. Compared with the model group， the GGQL and its combination groups showed improved pathological injury and morphological structure of colon tissues in UC rats， reduced serum levels of MPO， LPO， and MDA （P<0.05）， potentiated T-SOD activity （the PLR-absent GGQL group）， CAT activity （the GR-absent GGQL group and the SR-CR group）， and GSH activity （P<0.01）， and increased mRNA and protein expression of Nrf2， NQO1， and HO-1 in colon tissues. The difference in the GGQL group was significant （P<0.05）.ConclusionGGQL has a restorative effect on the pathological injury of UC rats， and its mechanism may be related to the activation of the Nrf2 signaling pathway and inhibition of oxidative stress response. The absence of PLR or only presence of SR and CR has a great impact on the treatment of UC. The results can provide references for the clinical rational medication of Chinese medicine and the research on the mechanism of compound combinations.Keywords：Gegen Qinliantang;different combinations;oxidative stress;ulcerative colitis;nuclear factor-erythroid 2 related factor （Nrf2） signaling pathway- 47
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Huanglian Jiedutang Regulates Inflammatory Immunity to Relieve Rheumatoid Arthritis
Abstract：ObjectiveTo observe the effects of Huanglian Jiedutang on pathological and immune damage in collagen-induced arthritis （CIA） model mice， and to explore the possible mechanism of Huanglian Jiedutang in relieving rheumatoid arthritis.MethodTwenty-four DBA/1 mice were randomly divided into normal group， model group， methotrexate group and Huanglian Jiedutang group， with six mice in each group. The CIA mice model were established using type Ⅱ collagen induction. The administration groups were respectively treated with Huanglian Jiedutang （5 g·kg-1） and methotrexate （0.5 mg·kg-1）. The joint swelling symptoms of the mice were observed， and the arthritis index was scored every 3 days. Flow cytometry was employed to detect granulocytes， monocytes， and T lymphocytes in peripheral blood. The expression of inflammatory cytokines in joint was determined by real-time polymerase chain reaction （Real-time PCR）. The ankle joint was scanned by micro-computed tomography （Micro-CT）， and the histopathological changes were observed through hematoxylin-eosin （HE） staining.ResultCompared with the normal group， the modeling led to joint swelling， elevated the joint index score （P<0.05）， increased the proportion of granulocytes （P<0.05） and decreased that of monocytes and T lymphocytes （P<0.01） in peripheral blood， and raised the neutrophil-to-lymphocyte ratio （NLR） （P<0.01）. Further， it up-regulated the expression of inflammatory cytokines such as tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， and IL-6 in joint （P<0.01）. Micro-CT showed obvious bone destruction in the ankle joint， and pathological examination revealed the infiltration of a large number of inflammatory cells and the synovial hyperplasia of joint tissue. Compared with the model group， Huanglian Jiedutang alleviated the symptoms of joint swelling， lowered the score of arthritis index （P<0.05）， increased the proportion of T lymphocytes and lowered NLR （P<0.01）. Moreover， it down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in joint （P<0.01） and alleviated the bone destruction and pathological injury of joint tissue.ConclusionType Ⅱ collagen caused systemic and local inflammatory immune damage in CIA mice. Huanglian Jiedutang alleviates such injury， especially for that in local joint， thereby inhibiting joint injury and bone destruction in CIA mice.Keywords：rheumatoid arthritis;Huanglian Jiedutang;collagen-induced arthritis （CIA） mouse;inflammatory immunity- 55
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Efficacy and Mechanism of Shenbai Jiedu Prescription Against Proliferation of HCT116 Cells
Abstract：ObjectiveTo investigate the mechanism by which Shenbai Jiedu prescription （SBJDF） inhibits the proliferation of colorectal cancer （CRC） HCT116 cells.MethodAfter 48 h treatment of HCT116 cells with SBJDF （0， 0.25， 0.5， 1， 2， 4 g·L-1）， the viability of HCT116 cells were determined by methyl thiazolyl tetrazolium （MTT） colorimetry. Following the classification of cells into blank control group and SBJDF （1， 2， 4 g·L-1） groups， the effect of SBJDF on HCT116 cell morphology was observed under an inverted microscope. The effects of SBJDF on the proliferation of HCT116 cells and mitochondrial membrane potential （Δψm） were detected by colony formation assay and JC-1 probe， respectively. The flow cytometry was then performed for determining cell cycle distribution and apoptosis. The effects of SBJDF on cell cycle-， apoptosis-， and nuclear factor kappa-B （NF-κB） signaling pathway-related proteins were determined by Western blot.ResultSBJDF effectively inhibited the vitality of HCT116 cells and changed their morphology in a concentration-dependent manner. Compared with the blank control group， SBJDF at 1， 2， 4 g·L-1 significantly reduced cell colony formation （P<0.05， P<0.01），and SBJDF at 2 and 4 g·L-1 arrested the HCT116 cell cycle at G0/G1 phase （P<0.05， P<0.01）. Compared with the blank control group， SBJDF at 1， 2， 4 g·L-1 remarkably down-regulated the protein expression of CyclinD1 （P<0.05， P<0.01）. SBJDF at 2 and 4 g·L-1 lowered the CyclinA2 and cyclin-dependent kinase 4 （CDK4） （P<0.05， P<0.01）. SBJDF at 4 g·L-1 reduced the cyclin-dependent kinase 1 （CDK1） （P<0.01）. Compared with the blank control group， SBJDF at 2 and 4 g·L-1 induced HCT116 cell apoptosis， down-regulated the protein expression of anti-apoptosis-related proteins Bcl-2 and Bcl-xl as well as the NF-κB signaling pathway-related proteins IκB kinase α （IKKα），inhibitor α of NF-κB （IκBα），and phospho-NF-κB p65 （p-p65） （P<0.05， P<0.01）， and diminished the mitochondrial membrane potential of HCT116 cells.ConclusionSBJDF inhibits the proliferation of HCT116 cells， which may be related to its inhibition of the activation of NF-κB signaling pathway and the induction of cell cycle arrest and apoptosis.Keywords：cancer poison;colorectal cancer （CRC）;apoptosis;cell cycle;nuclear factor kappa-B （NF-κB） signaling pathway- 31
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Activating Effect of Effective Components in Medicinal and Edible Substances on Human Pregnane X Receptor and Cytotoxicity Screening
Abstract：ObjectiveTo explore the activating effects of ten important effective components in seven medicinal and edible substances on human pregnane X receptor （PXR）， including Glycyrrhizae Radix et Rhizoma （liquiritin and glycyrrhizic acid）， Houttuyniae Herba （quercetin and houttuyfonate）， Prunellae Spica （rosmarinic acid）， Cassiae Semen （aurantio-obtusin）， Poria （pachymic acid）， Lilii Bulbus （Lilium brownii saponin and colchicine）， and Lycii Fructus （Lycium barbarum polysaccharide） and screen potentially toxic components.MethodCell counting kit-8 （CCK-8） assay was used to investigate the cytotoxic effect of liquiritin， glycyrrhizic acid， quercetin， houttuyfonate， rosmarinic acid， pachymic acid， aurantio-obtusin， and colchicine （10， 20， and 50 μmol·L-1）， and L. brownii saponin and L. barbarum polysaccharide （10， 20， and 50 mg·L-1） on normal human hepatocyte cell line （L02）. The release of lactate dehydrogenase （LDH） in L02 cells after drug treatments was detected by the biochemical analyzer. The apoptosis induced by ten effective components was explored by Hoechst 33342 staining. The secreted luciferase reporter system was used to co-transfect the PXR expression vector and reporter gene vector containing cytochrome P450 3A4 （CYP3A4） transcriptional regulatory region into L02 cells， with 10 μmol·L-1 rifampicin （RIF） as a positive control. After treated with liquiritin， glycyrrhizic acid， quercetin， houttuyfonate， rosmarinic acid， aurantio-obtusin， pachymic acid， and colchicine （5， 10， and 20 μmol·L-1） and L. brownii saponin and L. barbarum polysaccharide （5， 10， and 20 mg·L-1） for 24 h， the cells were tested for secreted luciferase activity.ResultCompared with the control group， colchicine， L. brownii saponin， and quercetin decreased the cell viability （P<0.05， P<0.01）. Compared with the control group， quercetin， rosmarinic acid， glycyrrhizic acid， colchicine， aurantio-obtusin， and pachymic acid increased the release rate of LDH in L02 cells （P<0.05， P<0.01）. The proportion of hyperchromatic nuclei increased gradually after rosmarinic acid， liquiritin， and L. barbarum polysaccharide treatments as compared with the control group （P<0.05， P<0.01）. In terms of co-transfection of pcDNA3.1-PXR and pGLuc-CYP3A4 into L02 cells， compared with the control group， aurantio-obtusin and pachymic acid showed activating effects on PXR （P<0.05）， whereas liquiritin and glycyrrhizic acid showed inhibitory effects （P<0.05）.ConclusionThe findings suggest that when medicinal and edible substances are taken for a long time， attention should be paid to their influence on drug-metabolizing enzymes and possible interactions， so as to improve their safety.Keywords：medicinal and edible substances;human pregnane X receptor;cytochrome P450 3A4 （CYP3A4）;secreted luciferase reporter system- 35
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Effect of Ginsenoside Rg1 on Biological Activity of Cryopreserved Schwann Cells and Nerve Regeneration After Allograft in Rat Sciatic Nerve
Abstract：ObjectiveTo observe the effect of ginsenoside Rg1 （G-Rg1） on the biological activity of cryopreserved Schwann cells （SCs） of the rat sciatic nerve and explore the feasibility of G-Rg1 in reducing the cryopreservation-induced injury in SCs.MethodBilateral sciatic nerves of SD rats were randomly divided into a fresh group， a blank group， and five G-Rg1 groups of different doses （1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1）. The nerves in the blank group and the G-Rg1 groups were preserved in liquid nitrogen solutions containing 0， 1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1 G-Rg1 for four weeks. The apoptosis of SCs was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）/S100 immunofluorescence staining. The expression of cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， major histocompatibility complex （MHC）-Ⅰ， and MHC-Ⅱ was detected by Western blot. Subsequently， all nerves were cultured in the incubator at 37 ℃ with 5% CO2 for 7 days. The expression of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF） was detected by Western blot. In addition， the above cryopreserved nerves in the blank group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 groups were transplanted to the Wistar rats by allografting （blank transplantation group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 transplantation groups）， and fresh sciatic nerve allograft and isograft control group were set up. Sixteen weeks after transplantation， compound muscle action potential （CMAP） and nerve conduction velocity （NCV） were measured by electrophysiology. Nerve filament （NF）200 immunofluorescence staining， transmission electron microscopy， and toluidine blue staining were used to analyze the histology of the regenerated nerves.ResultCompared with the fresh group， the blank group and the G-Rg1 groups showed increased expression of Caspase-9， Caspase-3， and the apoptosis of SCs （P<0.05，P<0.01） and decreased expression of GDNF， NGF， MHC-Ⅰ， and MHC-Ⅱ （P<0.01）. Compared with the results in the blank group， the expression of Caspase-9 and Caspase-3 decreased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups （P<0.01）， and the apoptosis of SCs was reduced in the 1×10-7-1×10-4 mol·L-1 G-Rg1 groups（P<0.05，P<0.01） and increased in the 1×10-3 mol·L-1 group （P<0.05）， while the expression of GDNF and NGF increased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups and decreased in the 1×10-3 mol·L-1 group （P<0.05）. There was no statistical significance in the expression of MHC-Ⅰ and MHC-Ⅱ between the blank group and the G-Rg1 groups. Compared with the 1×10-7 mol·L-1 and 1×10-3 mol·L-1 G-Rg1 groups， the 1×10-6 1×10-5， 1×10-4 mol·L-1 G-Rg1 groups showed decreased expression of Caspase-3 and the apoptosis of SCs （P<0.05，P<0.01） and increased expression of GDNF and NGF （P<0.05，P<0.01）. There was no statistical significance in MHC-Ⅰ and MHC-Ⅱ expression among G-Rg1 groups. Sixteen weeks after transplantation， compared with the isograft group， the blank transplantation group and the G-Rg1 transplantation groups showed decreased CMAP， NCV， myelin sheath thickness， and number of myelinated nerve fibers （P<0.01）， and the 1×10-6 and 1×10-4 mol·L-1 G-Rg1 transplantation groups showed decreased NF200 （P<0.01）. Compared with the allograft group， the blank transplantation group and the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Compared with the blank transplantation group， the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Among all groups of G-Rg1 transplantation， each index of the 1×10-5 mol·L-1 G-Rg1 transplantation group was superior to that of the 1×10-4 and 1×10-6 mol·L-1 G-Rg1 transplantation group （P<0.05）.ConclusionG-Rg1 at a certain centration can maintain the biological activity of cryopreserved SCs of rat sciatic nerve， alleviate the cryopreservation-induced injury of rat sciatic nerve， and promote nerve regeneration after allograft.Keywords：ginsenoside Rg1;peripheral nerve cryopreservation;cryodamage;allograft;nerve regeneration- 22
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Mechanism of Danggui Niantongtang on Adjuvant Arthritis Rats with Wind-dampness-heat Arthralgia Based on Quantitative Proteomics
Abstract：ObjectiveTo explore the mechanism of Danggui Niantongtang （DGNT） against adjuvant arthritis （AA） rats with wind-dampness-heat arthralgia by quantitative proteomics.MethodSixty SD rats were randomly divided into normal group， model group， angelica came pain soup low， medium and high dose group and methotrexate （MTX） group， each group of 10， only the rat tail root subcutaneously inactivated mycobacterium tuberculosis （Mtb） of adjuvant to build model of AA， artificial climate box intervention 16 d rheumatic fever bi syndrome model is set up， building the day began to drug intervention， The intervention lasted for 28 days. The proteins of synovial tissues in experimental rats were extracted. The differential proteins in the medium-dose DGNT group and the model group were detected and analyzed by 4D label-free quantification （4D-LFQ） proteomics. The differentially expressed proteins associated with mitochondrial pathway apoptosis were verified by immunohistochemistry and Western blot.ResultA total of 4 756 proteins were identified from rat synovial tissues， of which 4 234 proteins contained quantitative information. There were 814 differential proteins between the model group and the DGNT group. As revealed by Gene Ontology （GO） and Kyoto Encyclopedia of Gene and Genome （KEGG） enrichment analyses， DGNT had an effect on the synovial proteome of AA rats with wind-dampness-heat arthralgia， and the differential proteins were enriched in the regulation of the immune system， response to acute inflammation， and apoptosis regulation. As demonstrated by the results of immunohistochemistry and Western blot， compared with the model group， the DGNT groups and the MTX group showed increased protein expression of B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax） and cytochrome C （Cyt C）（P<0.05， P<0.01）， reduced Bcl-2 level （P<0.05， P<0.01）， elevated level of cleaved cysteinyl aspartate-specific protease 9 （Caspase-9）/Caspase-9 （P<0.01）， and decreased level of phosphorylated protein kinase B （p-Akt）/Akt（P<0.05， P<0.01）.ConclusionDGNT involved multiple targets in the treatment of AA with wind-dampness-heat arthralgia and it may exert its effect in the prevention and treatment by regulating the Akt/Bax/Bcl-2 pathway and promoting the cell apoptosis in the mitochondrial pathway.Keywords：Danggui Niantongtang;wind-dampness-heat arthralgia;rheumatoid arthritis;proteomics;apoptosis- 30
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Coptidis Rhizoma Crude Polysaccharide and Berberine Synergistically Restore Intestinal Mucosal Barrier Damage in Ulcerative Colitis
Abstract：ObjectiveTo investigate the synergistic effect of Coptidis Rhizoma crude polysaccharide （CCP） and berberine （BBR） in treating ulcerative colitis （UC） model mice.MethodThirty male BALB/c mice were randomized into five groups. Except the 6 mice in the normal group， the rest were given 5% dextran sodium sulfate in their daily drinking water to establish the UC model. After modeling， the mice were administrated with corresponding agents by gavage once daily for 4 days： BBR （100 mg·kg-1） group， BBR （100 mg·kg-1） + low-dose （22.8 mg·kg-1） CCP group， BBR （100 mg·kg-1） + high-dose （45.6 mg·kg-1） CCP group. The mice in the model group and normal group were administrated with the same volume of normal saline. At the end of the experiment， the mice were sacrificed for the collection of colon， and the expression of tight junction proteins zonula occluden-1 （ZO-1）， Claudin-1， and Occludin in colon tissue was detected by Western blot. With the normal group as the control， the disease activity index （DAI） score， colon length， colon histomorphology， and expression levels of tight junction proteins in other groups were evaluated.ResultCompared with the normal group， the modeling down-regulated the protein levels of ZO-1， Claudin-1， and Occludin （P<0.01）. Compared with the model group， BBR did not significantly change the protein level of Claudin-1 and up-regulated those of ZO-1 and occludin （P<0.01）. The expression levels of Claudin-1， ZO-1， and Occludin were up-regulated in BBR + CCP groups （P<0.01）. The expression levels of tight junction proteins in BBR + CCP groups were significantly higher than those in the BBR group （P<0.05）.ConclusionThe administration of CCP combined with BBR can effectively ameliorate intestinal mucosal barrier damage in the mice with UC.Keywords：Coptidis Rhizoma crude polysaccharide;berberine;ulcerative colitis;tight junction proteins- 38
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Effect and Mechanism of Steamed Allii Sativi Bulbus Polysaccharide Against Dysbiosis-induced Diarrhea in Young Rats
Abstract：ObjectiveTo verify the efficacy of steamed Allii Sativi Bulbus polysaccharide in regulating the intestinal flora of young dysbiosis-induced diarrhea rats based on 16S rRNA gene sequencing.MethodThe young SD rats were randomly divided into blank group，model group，positive drug group （bifid triple viable capsules），and high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups，six in each group. The dysbiosis-induced diarrhea rat model was established，and the blank group and the model group were given normal saline by gavage （each 10 mL·kg-1），and the high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups were administered with corresponding drugs （500 mg·kg-1 and 250 mg·kg-1， respectively） ，once a day for seven consecutive days. The loose stool rate，loose stool grade，diarrhea index，small intestine propulsion rate and hematoxylin-eosin （HE） staining were used as indexes to investigate the effect of steamed Allii Sativi Bulbus polysaccharide on improving diarrhea symptoms in young rats. The feces of rats were collected for 16S rRNA gene high-throughput sequencing.ResultCompared with the model group， the positive drug group and the high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups had alleviated symptoms， down-regulated loose stool rate and diarrhea index （P<0.01） and decreased small intestine advancement rate （P<0.05）. HE staining showed that after the treatment with steamed Allii Sativi Bulbus polysaccharide，the inflammatory cell infiltration of the colon tissue was improved and the intestinal gland recovered to the normal condition，which indicated that steamed Allii Sativi Bulbus polysaccharide could significantly ameliorate the diarrhea in young rats. The sequencing results revealed that steamed Allii Sativi Bulbus polysaccharide had a moderating effect on the abundance of the intestinal flora of young dysbiosis-induced diarrhea rats，elevating the flora richness and diversity indexes. Specifically， the abundance of Bacteroidota was increased while that of Firmicutes and Proteobacteria was decreased.ConclusionSteamed Allii Sativi Bulbus polysaccharide can be used to treat dysbiosis-induced diarrhea in young rats by regulating the abundance of intestinal microbiota.Keywords：Allii Sativi Bulbus polysaccharide;steaming;diarrhea;efficacy;mechanism- 23
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Xiaoai Jiedu Prescription-containing Serum Enhances Lethal Effect of NK Cells on Colon Cancer Cells
Abstract：ObjectiveTo explore the influence of Xiaoai Jiedu prescription （XJP）-containing serum on natural killer （NK） cells′ lethal effect on colon cancer cells and the molecular mechanism.MethodXJP-containing serum （0.1%， 0.5%， 1%， 5%， 10%） was used to treat HCT-116 cells and NK-92MI cells respectively for 24 h， and methyl thiazolyl tetrazolium （MTT） assay was employed to detect cell proliferation. Then， low-concentration （0.1%， 0.5%， 1%） XJP-containing serum was selected to treat co-cultured HCT-116 cells and NK-92MI cells for 24 h and calcein acetoxymethyl ester/propidium iodide （Calcein-AM/PI） was applied to detect the killing effect of NK cells on colon cancer cells. Flow cytometry was used to detect apoptosis of colon cancer cells， Western blot the expression of apoptosis-related proteins and signal transducer and activator of transcription 4 （STAT4） pathway-related proteins， and enzyme-linked immunosorbent assay （ELISA） the secretion of interferon （IFN）-γ.ResultHigh-concentration （5%， 10%） XJP-containing serum inhibited the proliferation of HCT-116 and NK-92MI cells （P<0.01）， while low-concentration （0.1%， 0.5%， 1%） XJP-containing serum had no obvious influence on cell proliferation compared with the blank group. As compared with the blank group， low-concentration XJP-containing serum enhanced the killing activity of NK cells against colon cancer cells in a concentration-dependent manner （P<0.01）， and induced apoptosis of colon cancer cells （P<0.01）. Moreover， XJP-containing serum （0.1%， 0.5%， 1%） down-regulated the expression of B-cell lymphoma 2 （Bcl-2） and B-cell lymphoma-extra large （Bcl-xl）， and up-regulated the expression of Bcl-2-associated X （Bax） compared with the blank group （P<0.05， P<0.01）. Compared with the co-culture group， XJP-containing serum （0.1%， 0.5%， 1%） increased the expression of p-STAT4 and IFN-γ （P<0.05）. ELISA result showed that XJP-containing serum （0.1%， 0.5%， 1%） raised IFN-γ secretion （P<0.01）.ConclusionXJP-containing serum can enhance the activity of NK cells to kill colon cancer cells. The mechanism is the likelihood that it activates STAT4 pathway， increases IFN-γ secretion by NK cells， down-regulates the expression of Bcl-xl and Bcl-2， and up-regulates the expression of Bax， thereby promoting the apoptosis of colon cancer cells.Keywords：Xiaoai Jiedu prescription;colon cancer;NK cells;apoptosis- 33
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Effect and Mechanism of Berberine Hydrochloride Against Lung Cancer Cells
in Vitro Based on Mevalonate PathwayAbstract：ObjectiveTo investigate the efficacy and mechanism of berberine hydrochloride （BBH） against lung cancer cells through the mevalonate （MVA） pathway.MethodHuman lung cancer A549 cells and mouse Lewis lung carcinoma （LLC） cells were used as research subjects. Cell proliferation and cell counting kit-8 （CCK-8） assay were performed to detect the inhibitory effect of BBH （10， 20， 30， 40， 50 μmol·L-1） on the proliferation of the two kinds of cells （48 h）. Then cell scratch assay was used to explore the influence of BBH （40 μmol·L-1） on the migration of A549 and LLC cells （24， 48 h）， and colony formation assay was conducted to compare the colony formation ability of the cells under different concentrations of BBH （10， 20， 40 μmol·L-1）. Moreover， the effects of BBH （40 μmol·L-1） on the content of acetyl-coenzyme A （A-CoA） and total cholesterol （TC） in A549 and LLC cells were determined by kit assay. AutoDock Vina was used for the dock of BBH and MVA pathway regulatory protein， sterol regulatory element-binding protein 2 （SREBP2）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to observe the effects of BBH （40 μmol·L-1） on the mRNA expression of nine genes related to the MVA pathway in A549 and LLC cells： hydroxymethylglutaryl-CoA synthase 1 （HMGCS1）， hydroxymethylglutaryl-CoA Reductase （HMGCR）， mevalonate kinase （MVK）， phosphomevalonate kinase （PMVK）， mevalonate 5-pyrophosphate decarboxylase （MVD）， farnesyl diphosphate synthase （FDPS）， squalene epoxidase （SQLE）， farnesyl-diphosphate farnesyltransferase 1 （FDFT1）， and geranylgeranyl diphosphate synthase 1 （GGPS1）. Western blot was performed to clarify the effects of BBH （40 μmol·L-1） on the expression of three key proteins of the MVA pathway： HMGCS1， HMGCR， and FDFT1. The Cancer Genome Atlas （TCGA） database was searched to analyze the relationship between HMGCS1， HMGCR， FDFT1 and transcription gene SREBF2 in non-small cell lung cancer （NSCLC）.ResultCompared with the conditions in the control group， the proliferation， migration， and colony formation of A549 and LLC cells in the BBH group were decreased （P<0.01）， while the cell apoptosis rate was increased （P<0.01）. Molecular docking showed that BBH had good binding activity with SREBP2. In addition， the content of A-CoA and TC of the MVA pathway was reduced （P<0.01）. BBH down-regulated the mRNA expression of HMGCS1， HMGCR， MVK， PMVK， MVD， FDPS， SQLE， FDFT1， and GGPS1 in A549 and LLC cells （P<0.01）， and lowered the levels of HMGCS1， HMGCR， and FDFT1 proteins （P<0.05， P<0.01）. In NSCLC patients， HMGCS1， HMGCR， and FDFT1 were highly correlated with SREBF2 （R=0.54， R=0.57， and R=0.48）.ConclusionBBH can inhibit the proliferation， migration， and colony formation of A549 and LLC cells and promote cell apoptosis， which may be related to the regulation of MVA pathway by BBH binding to SREBP2.Keywords：berberine hydrochloride;lung cancer cells;molecular docking;mevalonate pathway- 25
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Renal Protective Effect of Bupi Yishen Decoction on Diabetic Kidney Disease Rats Based on Tubuloglomerular Feedback
Abstract：ObjectiveTo observe the influence of Bupi Yishen decoction on the curative effect of diabetic kidney disease rats and the tubuloglomerular feedback.MethodAmong the 100 SD male rats， 25 were randomly selected as the normal group， and the remaining 75 were treated with high-fat and high-sugar diet combined with intraperitoneal injection of low-dose streptozotocin （STZ） to establish diabetic rat model. The model rats were randomly divided into model group， Bupi Yishen group and losartan group. Bupi Yishen group and Losartan group were given Bupi Yishen decoction （23.4 g·kg-1） and losartan tablet （9 mg·kg-1）， respectively by gavage， and normal group and model group received equal volume of distilled water via intragastric administration. The treatment lasted for 11 consecutive weeks. Random blood glucose was measured once every two weeks. Rats in each group were put into metabolic cage and 24-hour urine volume was recorded. At the end of week 11， the rats were sacrificed for index detection. Enzyme linked immunosorbent assay （ELISA） was used to determine the quantification of 24-hour urinary microalbumin excretion rate （24 h UAER）， renal tubular injury-related proteins and urine creatinine. Blood was collected for creatinine and urea nitrogen detection. Hematoxylin-eosin （HE） staining， periodic acid-schiff （PAS） staining and Masson staining were performed to observe the pathological changes of the kidney of rats. The expression of sodium-glucose cotransporter 1 （SGLT1）， sodium-glucose cotransporter 2 （SGLT2）， sodium-potassium-chloride cotransporter 2 （NKCC2） and connexin 40 （CX40） was detected by immunohistochemistry. The location of adenosine type Ⅰ receptor （A1AR） in kidney was observed by immunofluorescence. In addition， the expression of SGLT1， SGLT2， NKCC2， CX40 and A1AR in renal tissues was determined by Western blot and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）.ResultCompared with normal group， renal pathology in model group showed increased glomerular volume， thickened basement membrane， and shed renal tubule epithelial cells. Compared with the model group， the renal pathology of bupiyishen group was improved. Compared with normal group， random blood glucose at different time points， 24 h UAER and renal tubule-injury-related proteins were increased in model group （P<0.01）， and the expressions of SGLT1 and SGLT2 in renal tissue were increased （P<0.01）， and the expression of NKCC2， CX40 and A1AR decreased （P<0.01）. Compared with the model group， the random blood glucose， 24 h UAER and renal tubular injury-related proteins in Bupi Yishen group decreased （P<0.01）， the expressions of SGLT1 and SGLT2 were down-regulated， and the expressions of NKCC2， CX40 and A1AR were increased （P<0.05）.ConclusionBupi Yishen decoction can improve glomerular and renal tubular injury and alleviate early hyperfiltration in diabetic kidney disease rats. The intervention effect of Bupi Yishen decoction on diabetic kidney disease is related to the regulation of tubuloglomerular feedback.Keywords：diabetic kidney disease;Bupi Yishen decoction;early hyperfiltration;tubuloglomerular feedback;adenosine type Ⅰ receptor- 21
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Fangji Huangqi Detumescence Prescription Treats Synovitis in Rats with Knee Osteoarthritis by Modulating Polarization of Synovial Macrophages
Abstract：ObjectiveTo assess the curative effects of Fangji Huangqi detumescence prescription （FHDP） on synovitis and polarization of synovial macrophages of knee osteoarthritis （KOA） model in rats induced by Hulth method.MethodThirty-six rats were randomly divided into sham operation group， model group， high-dose， medium-dose， and low-dose （29.16， 14.58， and 7.29 g·kg-1） FHDP groups， and loxoprofen sodium （16.2 mg·kg-1） group. KOA model in rats was induced by modified Hulth method. Six weeks after the operation， rats were given high， medium， and low concentrations of FHDP， normal saline （NS）， and loxoprofen sodium according to the group to intervene， and sacrificed after 2-week administration. Synovium and cartilage histopathological changes were observed after hematoxylin-eosin （HE） staining. Flow cytometry （FCM） and immunofluorescence （IF） test were used to evaluate the polarization of M1/M2 macrophages. Immunohistochemistry （IMC） and enzyme-linked immunosorbent assay （ELISA） were used to detect the related protein expression levels of macrophage polarization， such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， interleukin-10 （IL-10）， and matrix metalloproteinase-13 （MMP-13） in joint tissues and serum.ResultCompared with the sham operation group， Krenn and Mankin scores in the model group were significantly increased （P<0.01）. Compared with the model group， Krenn score was decreased in all administration groups （P<0.05， P<0.01）， but there was no significant difference in Mankin score in any administration groups. Compared with the sham operation group， M1/mø （CD38+） ratio in the model group was significantly increased （P<0.01）， and M2/mø （CD206+） ratio in the model group was decreased （P<0.05）. Compared with the model group， M1/mø ratio in the high， medium， and low-dose FHDP groups was decreased （P<0.05， P<0.01）， but M2/mø ratio was increased in all administration groups （the difference had no statistical significance）. Compared with the sham operation group， M1/M2 ratio in the model group was significantly increased （P<0.01）. Compared with the model group， M1/M2 ratio in all FHDP groups was significantly decreased （P<0.01）， and M1/M2 ratio in the high and medium-dose FHDP groups was lower than that in the loxoprofen sodium group （P<0.05）. Compared with the sham operation group， the levels of TNF-α， IL-1β， and MMP-13 in synovium and cartilage of the model group were significantly increased （P<0.01）， the level of IL-10 was significantly decreased （P<0.01）. Compared with the model group， the levels of TNF-α and IL-1β in synovium were decreased in all administration groups （P<0.05）， but the difference of the levels of MMP-13 and IL-10 in synovium had no statistical significance. The level of inflammatory mediators in cartilage was not affected in all administration groups. Compared with the sham operation group， the levels of TNF-α and IL-β in serum of the model group were significantly increased （P<0.01）， the level of IL-10 was decreased （P<0.05）. Compared with the model group， the level of TNF-α in the high-dose FHDP group was decreased （P<0.05）， and the level of IL-10 was increased in all administration groups （P<0.05， P<0.01）. The difference of the level of IL-β in all administration groups had no statistical significance.ConclusionFHDP attenuated the synovitis of KOA rats. FHDP exert the effect on the releasing of proinflammatory cytokines and MMP by inhibiting the polarization of M1 macrophages in synovium， and had no significant effect on the polarization of M2 macrophages. Modulating the imbalanced polarization of synovial macrophages was a possible mechanism of FHDP on attenuating synovitis and treating KOA.Keywords：Fangji Huangqi detumescence prescription （FHDP）;osteoarthritis;polarization of macrophages;synovitis;inflammation- 25
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Clinical Efficacy of Gandouling Combined with Short-term Decoppering on Hepatic Fibrosis of Wilson's Disease Based on Transient Elastography
Abstract：ObjectiveTo explore the accuracy of clinical common serum fibrosis indexes hyaluronic acid （HA）， type Ⅳ collagen （CⅣ）， laminin （LN）， and type Ⅲ procollagen peptide （PⅢNP）， in combination with liver stiffness measurement （LSM， measured by transient elastography） and non-invasive markers of fibrosis aspartate aminotransferase to platelet ratio index （APRI） and fibrosis-4 （FIB-4） in the prediction of the hepatic fibrosis of Wilson's disease （WD） and to observe the clinical effect of Gandouling （GDL）.MethodThe data of 76 WD patients were collected and the LSM， serum fibrosis indexes （HA， PⅢNP， CⅣ， LN）， APRI， and FIB-4 before treatment were recorded. The correlation of LSM with serum fibrosis indexes， APRI， and FIB-4 was discussed via Pearson′s correlation analysis. According to the therapeutic schemes， patients were classified into the control group （36 cases） and treatment group （40 cases）. Patients in control group were treated with sodium dimercaptopropylsulfonate （DMPS）， while those in the treatment group received GDL in addition to the western medicine therapy. The treatment lasted 6 courses （8 days/course） and the influence of GDL on the indictors was evaluated.ResultHA， CⅣ， LN， PⅢNP， APRI， and FIB-4 were in positive correlation with LSM （r=0.517， 0.438， 0.281， 0.457， 0.778， 0.847， P<0.01）. HA， CⅣ， LN， and PⅢNP in the treatment group were lower after treatment than before treatment （P<0.05， P<0.01）. HA， CⅣ， and LN in the control group were lower after treatment than before treatment （P<0.05， P<0.01）， and PⅢNP showed no significant difference. LSM， FIB-4， and APRI in both groups decreased after treatment （P<0.05）. After treatment， LSM， FIB-4， APRI， HA， and PⅢNP in the treatment group were lower than those in the control group （P<0.05， P<0.01）， but CⅣ and LN demonstrated no significant difference from the control group.ConclusionLSM in combination with serum fibrosis indexes （HA， PⅢNP， CⅣ， LN）， FIB-4， and APRI can help accurately identify the level of the hepatic fibrosis in WD. Moreover， on the basis of decoppering by western medicine， GDL can significantly improve the liver function and hepatic fibrosis of WD patients.Keywords：hepatic fibrosis;hepatolenticular degeneration;Gandouling;transient elastography;liver stiffness measurement;serum fibrosis index- 19
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Multi-dimensional Efficacy Analysis of TDP Combined with Osteoking in Treatment of Knee Osteoarthritis of Qi Stagnation and Blood Stasis Type
Abstract：ObjectiveTo evaluate the clinical efficacy of TDP （specific electromagnetic wave） combined with Osteoking in the treatment of knee osteoarthritis of Qi stagnation and blood stasis type.MethodA total of 104 patients with knee osteoarthritis of Qi stagnation and blood stasis type， who received conservative therapy in The Third People's Hospital of Xinjiang Uygur Autonomous Region from July 2019 to December 2021， were randomized into the control group and study group with the random number table method， 52 cases in either group. The control group was treated with TDP， and the study group with TDP and Osteoking. The treatment lasted 1 week for both groups， with 1-month follow-up. Subjective indexes of visual analog scale （VAS） score and Western Ontario McMaster Universities Osteoarthritis Index （WOMAC） score， and objective indexes of visual tenderness index and visual knee range of motion were determined before and after treatment to evaluate the pain and functions of patients. Traditional Chinese medicine （TCM） syndrome score was calculated. The serum erythrocyte sedimentation rate （ESR） and high sensitivity C-reactive protein （hs-CRP） were detected before and after treatment， and the total clinical effective rate was calculated.ResultBefore treatment， the baseline information and all the scores of the two groups were comparable. After treatment， the VAS score， WOMAC score， tenderness index， knee range of motion， and TCM syndrome score were improved in both groups （P<0.01）. After the treatment， the VAS score and WOMAC score of the study group were lower than those of the control group （P<0.01） and the improvement of tenderness index in the study group was better than that in the control group （P<0.05）. The knee range of motion in the study group was better than that in the control group （P=0.061）. The TCM syndrome score of study group was lower than that of control group （P<0.01） after treatment. The post-treatment serum ESR and hs-CRP level in the two groups decreased significantly after treatment， and the study group were lower than those of the control group （P<0.01）. The total clinical effective rate of the study group was 90.4%（47/52）， as compared with the 53.8%（28/52） in the control group （P<0.05）. No obvious adverse events occurred during treatment in both groups.ConclusionThe clinical efficacy of TDP combined with Osteoking in the treatment of knee osteoarthritis of Qi stagnation and blood stasis type is remarkable， which can improve knee pain and functions， alleviate TCM syndrome， and reduce inflammatory indexes， with high safety.Keywords：specific electromagnetic wave;Osteoking;Qi stagnation and blood stasis;knee osteoarthritis- 24
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Therapeutic Observation of Shipiyin Combined with Diosmin in Treatment of Lymphedema with Spleen Yang Deficiency Syndrome After Modified Radical Mastectomy
Abstract：ObjectiveTo investigate the clinical effect of Shipiyin combined with diosmin in the treatment of lymphedema with spleen Yang deficiency syndrome（SYDS）after modified radical mastectomy and the specific effect on the function of the affected limb.MethodEighty-two patients with lymphedema with SYDS after modified radical mastectomy from outpatient and inpatient department of breast department and oncology department of the First Affiliated Hospital of Hunan University of Chinese Medicine were randomly divided into an observation group（41 cases） and a control group（41 cases）. The control group was given diosmin tablets（0.9 g per time， two times per day）on the basis of conventional treatment，and the observation group was given Shipiyin（one dose per day）on the basis of the control group. The course of treatment was 14 days. The clinical symptoms were observed and the limb circumference，traditional Chinese medicine（TCM） syndrome score，functional assessment of cancer therapy-breast cancer（FACT-B） score，disability of arm， shoulder and hand questionnaire（DASH） score，and joint range of motion were measured to analyze the TCM syndrome therapeutic effect and clinical efficacy.ResultAfter 14 days of treatment， the total effective rate of the observation group was 85.37% （35/41） and that of the control group was 63.41% （26/41） in the TCM symptoms， showing a statistically significant difference （Z=-2.212， P<0.05）. In terms of the clinical efficacy， the total effective rate in the observation group was 82.93% （34/41） and that in the control group was 75.61% （31/41）， indicating a statistically significant difference （Z=-2.061， P<0.05）. Compared with the situations before treatment， the scores of clinical symptoms such as the swelling of the upper limb， pain， sense of heaviness， stuffiness， fatigue， fullness， tightness， and skin keratosis and pruritus in the two groups were significantly lowered （P<0.01） after treatment. Compared with the control group， the observation group could better improve the swelling and fullness（P<0.01），as well as the feeling of pain，heaviness，stuffiness，fatigue，tightness，skin keratosis and pruritus （P<0.05）of the upper limbs of patients. The affected limb circumference， TCM syndrome score， and DASH score decreased significantly， while the FACT-B score and upper limb joint range of motion increased significantly in the two groups after treatment （P<0.01）. Compared with the control group， the observation group showed significantly reduced limb circumference at 10 cm proximal to the elbow striae， lowered TCM syndrome score， elevated FACT-B score（P<0.05）， decreased DASH score and improved range of motion of the upper limb joint （P<0.01） after treatment.ConclusionShipiyin combined with diosmin has better clinical efficacy in the treatment of lymphedema with SYDS after modified radical mastectomy than diosmin alone，which can better improve the clinical symptoms，signs，quality of life and limb functional activity of patients. This provides a new clinical program for the treatment of lymphedema after breast cancer surgery with integrated Chinese and western medicine.Keywords：breast cancer;modified radical mastectomy;lymphedema;Shipiyin;diosmin;upper limb functional activity;traditional Chinese medicine- 20
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Wang's Yijing Decoction Combined with Conventional Therapy： Performance in Treating Polycystic Ovary Syndrome with Kidney Deficiency and Phlegm-dampness Syndrome and Regulating Effect on Intestine-brain Axis
Abstract：ObjectiveTo observe the therapeutic effect of Wang's Yijing decoction combined with conventional therapy on the treatment of polycystic ovary syndrome （PCOS） with kidney deficiency and phlegm-dampness syndrome and its regulating effect on the intestine-brain axis.MethodThe 132 patients from the gynecological clinic of Shanxi Bethune Hospital were randomly assigned into a control group and an observation group， with 66 patients in each group. The patients in the control group were treated with drospirenone and ethinylestradiol tablets and metformin tablets， and those in the observation group were administrated with Wang's Yijing decoction on the basis of the therapy in the control group. The treatment in both groups lasted for 3 menstrual cycles. The changes in menstrual blood volume， color， and texture， as well as that in the menstrual cycle before and after treatment were scored. Further， the Rosenfield， hirsutism， and kidney deficiency and phlegm-dampness syndrome were scored before and after treatment. Pelvic ultrasound was performed before and after treatment to compare the bilateral ovarian volume. The levels of anti-mullerian hormone （AMH）， estradiol （E2）， luteinizing hormone （LH）， follicle-stimulating hormone （FSH）， testosterone （T）， dihydrotestosterone （DHT）， and sex hormone-binding globulin （SHBG） were measured before and after treatment， and the free androgen index （FAI） was calculated. After the fasting insulin and fasting blood glucose were examined before and after treatment， the homeostatic model assessment of insulin resistance （HOMA-IR） was calculated. The levels of ghrelin， glucagon-like peptide-1 （GLP-1）， peptide tyrosine-tyrosine （PYY） in serum and short-chain fatty acids （SCFAs） in fresh stool were measured before and after treatment.ResultThe treatments in both groups lowered the scores of irregular menstruation， Rosenfield， hirsutism， and kidney deficiency and phlegm-dampness syndrome （P<0.01）， and the scores in the observation group were lower than those in the control group （P<0.01）. After treatment， the T， DHT， FAI， AMH， E2， LH， and FSH levels lowered （P<0.01）， while the SHBG level elevated （P<0.01）. In addition， the AMH， E2， LH， T， DHT， and FAI in the observation group were lower than those in control group， while the SHBG level presented an opposite trend （P<0.01）. After treatment， the bilateral ovarian volume and HOMA-IR of the two groups decreased （P<0.01）， and were lower in the observation group than in the control group （P<0.05， P<0.01）. The treatments in both groups elevated the levels of ghrelin， GLP-1， PYY， and SCFAs （P<0.01）， which were higher in the observation group than in the control group （P<0.01）. After treatment， the recovery rate in the observation group was 65.57% （40/61）， which was higher than 44.83% （26/58） in the control group （χ2=5.180， P<0.05）.ConclusionWang's Yijing decoction combined with conventional western medicine treatment can regulate the intestine-brain axis， endocrine and metabolic disorders， and sex hormones， adjust the menstrual cycle， and alleviate clinical symptoms， demonstrating good clinical effect for the PCOS patients with kidney deficiency and phlegm dampness syndrome.Keywords：polycystic ovary syndrome;kidney deficiency and phlegm-dampness syndrome;Wang's Yijing decoction;intestine-brain axis;intestine-brain peptides;short-chain fatty acids;insulin resistance- 22
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Discussion on Material Basis of Different Parts of Mulberry with Different Effects Based on Molecular Connectivity Index
Abstract：ObjectiveTo explore the material basis for the difference in the efficacy of different parts of mulberry based on molecular connectivity index （MCI）.MethodBy referring to the relevant literature at home and abroad and traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP） database， the chemical composition database of mulberry-source medicinal materials was established. Venn analysis was carried out on the components among mulberry-source medicinal materials. The components in the database were divided into 10 categories， and the composition information was analyzed. According to MCI value， all components of mulberry-source medicinal materials were divided into different groups. The angle cosine method was used to calculate the MCI similarity. The average MCI values of the common component group from 0-8 orders and CI of mulberry-source medicinal materials were calculated.ResultThe components with high similarity such as （+）-cycloolivil， 1′-methoxy-2′-hydroxydihydromollugin， kuwanon， morusin and 1-deoxynojirimycin were selected as potential pharmacodynamic components. Mulberry-source medicinal materials could be divided into five component groups. The similarity between component groups and total components was 0.760-0.999， and the similarity between component groups was 0.248-0.999. In Mori Ramulus， Mori Folium， Mori Cortex and Mori Fructus， the average MCI values of their flavonoids from 0-8 orders were 4.57， 4.59， 6.41， 4.24， respectively. The average MCI values of alkaloids from 0-8 orders were 2.65， 4.55， 2.58， 2.78， respectively. The average CI values from 0-8 orders were 5.51， 5.49， 5.44 and 2.88， respectively.ConclusionIt is preliminarily concluded that there are differences in the flavonoids and pathways of hypoglycemic effects between Mori Cortex and the other three mulberry-source medicinal materials. The MCI values of alkaloids from 0-8 orders in Mori Folium and Mori Fructus were higher， but their inhibitory activity of α-glucosidase were lower than those of Mori Ramulus and Mori Cortex. The structural characteristics of the total components of Mori Fructus represented by CI were quite different from the other three mulberry-source medicinal materials.Keywords：mulberry;mulberry-source medicinal materials;homologous and heterologous effects;molecular connectivity index;core index;quality marker;material basis- 41
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Establishment of Cryosectioning Method and Histochemical Localization Analysis of Volatile Oil and Coumarin Components of
Ferula ferulaeoides Abstract：ObjectiveTo establish the frozen section method of Ferula ferulaeoides， and to study the histochemical localization of volatile oil and coumarins in different organs of F. ferulaeoides.MethodThe roots， stems， petioles and leaves of F. ferulaeoides were used as materials to investigate the concentration of sucrose protectant， liquid nitrogen flash-freezing time， embedding conditions， section thickness， freezing temperature and time and post-treatment methods， the most suitable section conditions were screened by comparing the integrity， microscopic effect， elongation and clarity of frozen sections. Sudan Ⅳ staining method and fluorescence microscopy were used to locate the volatile oil and coumarins of F. ferulaeoides.ResultThe optimal conditions for frozen sections of the roots， stems， petioles and leaves of F. ferulaeoides were as follows：10%， 15% and 20% gradient sucrose as the protectant for roots， 10%， 20% and 30% gradient sucrose as the protectant for stems and petioles， 20%， 25% and 30% gradient sucrose as the protectant for leaves， glue-water （2∶1） as the embedding agent， quick-freeze in liquid nitrogen for 20 s， warmed up at -25 ℃ for 30 min， sliced at -20 ℃ with the thickness of 25 μm， rinsed with the same concentration of sucrose solution （gradient sucrose solution selected the last concentration）， and the slices placed on the ice pack for a period of time and stored at room temperature. Among them， the concentration of sucrose protectant was the most important factor. The results of histochemical localization showed that volatile oil and coumarins in four organs of F. ferulaeoides were mainly distributed in resin canal.ConclusionFrozen section of F. ferulaeoides is established for the first time with high rate of slicing and simplified steps， its volatile oil and coumarins are mainly accumulated in resin canal.Keywords：Ferula ferulaeoides;frozen section;volatile oil;coumarin;resin canal;sucrose- 16
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Rapid Identification of Gastrodiae Rhizoma with Different Sulfur Fumigation Levels Based on Ultra-fast Gas Phase Electronic Nose
Abstract：ObjectiveIn order to find a fast odor-based method for the identification of sulfur fumigated Gastrodiae Rhizoma， an ultra-fast gas phase electronic nose technology was used to identify the odors of different degrees of sulfur fumigated Gastrodiae Rhizoma decoction pieces.MethodHeracles NEO ultra-fast gas phase electronic nose was employed to collect gas chromatograms of unsulfured and sulfured with different degrees of Gastrodiae Rhizoma decoction pieces， gas chromatograms were performed under programmed temperature （initial temperature of 40 ℃， 0.2 ℃·s-1 to 60 ℃， and then 4 ℃·s-1 to 250 ℃）， the sample volume was 5 mL， the incubation temperature was 65 ℃ and incubation time was 35 min. Kovats retention index and the AroChemBase database were used for qualitative analysis， and stoichiometric analysis was performed on this basis. Principal component analysis （PCA）， discriminant factor analysis （DFA） and partial least squares-discriminant analysis （PLS-DA） models were established to identify the Gastrodiae Rhizoma decoction pieces with different degrees of sulfur fumigation.ResultAccording to the comparative analysis of AroChemBase database， there were significant differences in the odor characteristics of sulfur fumigated and non-sulfur fumigated Gastrodiae Rhizoma， cyclopentane， acetone and heptane might be the odor components to distinguish the degree of sulfur fumigation in Gastrodiae Rhizoma decoction pieces. The identification index of PCA model was 81， the accumulative discriminant index of the discriminating factors was 92.09% in DFA model， the supervisory model interpretation rate of PLS-DA model was 0.963 and the predictive ability parameter was 0.956， indicating that PCA， DFA and PLS-DA models could well distinguish Gastrodiae Rhizoma decoction pieces with different sulfur fumigation degrees.ConclusionHeracles NEO ultra-fast gas phase electronic nose can be used as a rapid method to identify and distinguish Gastrodiae Rhizoma decoction pieces with different levels of sulfur fumigation. Meanwhile， it can provide a rapid， simple and green method and technology for identification of traditional Chinese medicine decoction pieces by sulfur fumigation.Keywords：Gastrodiae Rhizoma;sulfur fumigation;ultra-fast gas phase electronic nose;cyclopentane;acetone;chemical pattern recognition methods;volatile component- 16
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Molecular Identification of Tibetan
Codonopsis Plants Based on ITS2 andpsbA-trnH BarcodeAbstract：ObjectiveTo identify the molecular biology of various species of Tibetan Codonopsis plants based on internal transcribed spacer（ITS）2 and psbA-trnH sequence barcode technology.MethodThe genomic DNA of 28 Tibetan Codonopsis plant samples from four species （Codonopsis canescens，C. foetens subsp. nervosa，C. pilosula， and C. thalictrifolia var. mollis） were extracted，and the ITS2 and psbA-trnH sequences were amplified and sequenced. The related sequences of 81 Tibetan Codonopsis plant samples belonging to 15 species were downloaded from GenBank， and MEGA 6.0 was used for sequence comparison and mutation site analysis. The GC content and genetic distance within and between species were calculated. Additionally， phylogenetic trees were constructed by maximum likelihood （ML） method， neighbor-joining （NJ） method，and unweighted pair-group method with arithmetic means （UPGMA） .ResultAccording to the mutation site，C. canescens， C. pilosula，C. pilosula subsp. tangshen， C. pilosula var. modesta，C. bhutanica，C. clematidea，C. lanceolata，C. subglobosa and C. foetens were distinguished. In the phylogenetic trees，the optimal clustering effects for ITS2 and psbA-trnH sequences were obtained using the ML method and the UPGMA method， respectively， and 12 species were effectively clustered.ConclusionITS2 and psbA-trnH sequences have a high identification rate for species of single origin，but there are still some limitations in identifying variants and original variants. This study provides basis for the identification of affinity relationship and clinical safety of Tibetan Codonopsis plants.Keywords：Codonopsis plants;DNA barcoding;internal transcribed spacer（ITS） 2 sequence;psbA-trnH sequence;molecular identification- 21
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Yield， Quality， and Soil Microecology of
Panax ginseng Under Different ForestsAbstract：ObjectiveTo explore the forest type and soil environment suitable for Panax ginseng.MethodThe yield， quality， soil chemical properties， soil enzyme activity， and soil microbial metabolism of 9-year-old P. ginseng under different forests were investigated.ResultThe quality of P. ginseng was significantly different among forest types. To be specific， P. ginseng under the Quercus mongolica forest had the highest quality， with the total saponin content of 2.27% which was 51.89% higher than that in P. ginseng under Larix gmelinii forest. The yield of P. ginseng under Q. mongolica forest and L. gmelinii forest （30 g·m-2） was the highest， 62.5% higher than that under Betula platyphylla forest. The soil content of organic matter， Cu， and Zn， and activity of sucrase and urease under Q. mongolica forest were lower than those under other forest types. The utilization rate of D-galacturonic acid by soil microorganisms under Q. mongolica forest was higher than that under other forest types， but the utilization rate of L-phenylalanine was lower than that under other forest types. The utilization rate of 2-hydroxybenzoic acid by soil microorganisms of B. platyphylla forest was significantly lower than that under other forest types. There was a negative correlation between soil Zn and ginsenoside Rb1 and Rc， and between soil K and ginsenoside Rb2 and Rb3. Mn and Cu were positively correlated with most saponins. The results of redundancy analysis showed that the soil microorganisms using carbon sources of amino acids， esters， acids， and sugars were the main factors causing the differences in P. ginseng among different forest types.ConclusionThe yield and quality of P. ginseng under Q. mongolica forest were the best， followed by the forest with different tree species， and coming in last was the B. platyphylla forest. This study is expected to provide theoretical support for the improvement of P. ginseng yield and quality and the improvement of ecological planting technology.Keywords：different forest types;understory Panax ginseng;soil microecology;soil enzyme activity;Biolog-Eco- 25
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
GC-MS Metabolites and Gene Expression Characteristics in Fibrous Roots of
Dioscorea zingiberensis in Response to Low Phosphorus Stress Enhanced PublicationAbstract：ObjectiveTo investigate the metabolites and gene expression characteristics in fibrous roots of Dioscorea zingiberensis in response to low phosphorus stress.MethodThe severe stress group， the moderate stress group， and the normal group were set up to stimulate the low phosphorus stress experiment. The fibrous roots of D. zingiberensis were collected during initial stress. The metabolites and transcriptomic characteristics were analyzed by gas chromatography-mass spectrometry （GC-MS） derivatization and RNA-seq techniques. Through multivariate statistical analysis of metabolites treated by different methods，functional analysis of differentially expressed genes， and data mining， the metabolism markers produced in fibrous roots of D. zingiberensis under low phosphorus stress were screened out， and the metabolic pathway characteristics of different genes were analyzed.ResultA total of 116 GC-MS metabolites were detected from the fibrous roots of D. zingiberensis. The metabolic characteristics of fibrous roots of D. zingiberensis under different low phosphorus treatments were obviously different. Orthogonal partial least squares discriminant analysis（OPLS-DA） model was used to screen six differential metabolites represented by sugars and alcohols from metabolites of fibrous roots treated with different methods，and these components were presumedly metabolism markers of fibrous roots of D. zingiberensis in response to low phosphorus stress. The differential genes screened out from the severe stress group and the normal group were mainly enriched in peroxidase pathway，phosphate and hypophosphate metabolism pathway，while the differential genes screened out from the severe stress group and the moderate stress group were mainly enriched in glutathione metabolism pathway and phosphopentose pathway. A total of 177 differential genes in response to low phosphorus stress were screened out from fibrous roots， involving many pathways such as terpenoid skeleton and inositol biosynthesis，which was consistent with the fact that the metabolic differential components in fibrous roots in response to low phosphorus stress were mainly saccharides and inositol.ConclusionThe metabolites and gene expression in fibrous roots of D. zingiberensis responded to low phosphorus stress，and the differential metabolites were closely related to differentially expressed genes. This study is expected to provide a theoretical basis for the research on the molecular mechanism of D. zingiberensis in response to low phosphorus stress.Keywords：Dioscorea zingiberensis;low phosphorus stress;gas chromatography mass spectrometry （GC-MS）;RNA-Seq;differential metabolites;differential genes- 20
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Predictive Analysis of Quality Markers of Jiangxi Aurantii Fructus Based on Fingerprint and Network Pharmacology Enhanced Publication
Abstract：ObjectiveTo analyze and predict the potential quality markers （Q-Marker） in the Genuine medicinal materials Jiangxi Aurantii Fructus based on fingerprints and network pharmacology.MethodUltra-high performance liquid chromatography （UPLC） and gas chromatography-mass spectrometry （GC-MS） fingerprints were established for 18 batches of Jiangxi Aurantii Fructus ，combined with chemometric methods to screen out candidate Q-Marker components.Use network pharmacology to construct a "core component-target-pathway" network to predict the Q-Marker and core targets of Jiangxi Aurantii Fructus，and then verify the biological activity of Jiangxi Aurantii Fructus Q-Marker by molecular docking method.ResultThe 18 batches of Jiangxi Aurantii Fructus use UPLC，GC-MS fingerprints combined with chemometric analysis，a total of 9 Q-Marker candidate components were screened out.Through network pharmacological analysis，it is predicted that nobiletin，neohesperidin，meranzin，naringin and D-limonene are the Q-Marker of Jiangxi Aurantii Fructus，acting on the core targets transforming protein p21/H-Ras-1（HRAS），cellular tumor antigen p53 （TP53），mitogen-activated protein kinase 8 （MAPK8），transcription factor AP-1（JUN），glycogen synthase kinase-3 beta（GSK3B），tumor necrosis factor（TNF），cyclin-dependent kinase inhibitor 1（CDKN1A），cAMP-dependent protein kinase catalytic subunit alpha（PRKACA），cysteine aspartate-specific protease-9（Caspase-9），cyclic AMP-responsive element-binding protein 1（CREB1），exerting gastrointestinal motility and antidepressant，anti-inflammatory，anti-tumor，etc.； molecular docking shows that nobiletin，neohesperidin，meranzin，naringin and D-limonene and the selected 10 core targets have good binding ability，reflecting the better biological activity of the Q-Marker of Jiangxi Aurantii Fructus.ConclusionThe Q-Marker of Jiangxi Aurantii Fructus can be comprehensively predicted from the two aspects of volatile and non-volatile components，providing a reference for the quality control of Jiangxi Aurantii Fructus and the further study of its pharmacodynamic mechanism.Keywords：Jiangxi Aurantii Fructus;quality marker;fingerprint;network pharmacology;molecular docking;nobiletin;neohesperidin;meranzin;naringin;D-limonene- 37
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Mechanism of Astragaloside Ⅳ in Treating Diabetic Retinopathy Based on Network Pharmacology and Molecular Docking Enhanced Publication
Abstract：ObjectiveTo reveal the pharmacological mechanisms of astragaloside Ⅳ（AS-Ⅳ）in treating diabetic retinopathy based on network pharmacology and molecular docking and to provide reference for new drug development and mechanism research.MethodPotential targets of AS-Ⅳ were obtained from SwissTargetPrediction and Targetnet. The targets of diabetic retinopathy were screened using GeneCards，Online Mendelian Inheritance in Man（OMIM） and Therapeutic Target Database. The targets of AS-Ⅳ and diabetic retinopathy were intersected by Venny 2.1.0. STRING platform and Cytoscape 3.7.2 were used to construct protein-protein interaction（PPI） network and screen core targets， respectively. Then，Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed. Furthermore，the binding affinity of AS-Ⅳ to key target receptors was assessed by molecular docking with Autodock Vina， and the key target signaling transduction pathway wasResultA total of 56 intersected targets of AS-Ⅳ and diabetic retinopathy were found，and the top five key targets were obtained through PPI network analysis：protein kinase B（Akt）1，vascular endothelial growth factor A（VEGFA），epidermal growth factor receptor（EGFR），Src and signal transducer and activator of transcription 3（STAT3）. Molecular docking verified the strong binding affinity of AS-Ⅳ to the five key target receptors. In addition，in vitro tests have been confirmed that AS-Ⅳ attenuated high glucose-induced injury in human retinal pigment epithelial cell line ARPE-19 by regulating Akt/Nrf2/HO-1 and Akt/glycogen synthase kinase-3β（GSK-3β）signaling pathways.ConclusionThere was a significant overlap in the targets of AS-Ⅳ and diabetic retinopathy. The key targets and pathways may reveal the main pharmacological mechanism of AS-Ⅳ in the treatment of diabetic retinopathy.Keywords：astragaloside Ⅳ;diabetic retinopathy;network pharmacology;molecular docking;molecular mechanism- 80
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Mechanism of Huanglian Houpotang on Ulcerative Colitis by Network Pharmacology and Experimental Verification Enhanced Publication
Abstract：ObjectiveTo explore the active components and underlying mechanism of Huanglian Houpotang （HHD） against ulcerative colitis（UC） based on network pharmacology and animal experiments.MethodThe active components of HHD were preliminarily obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM） and screened out by TCMSP， SwissADME， and SwissTargetPrediction， and their targets were predicted. Relevant microarrays were searched for disease genes with the help of Gene Expression Omnibus （GEO）. The common targets of HHD and disease genes were screened out to obtain the potential targets of HHD against UC. The drug-active component-target-disease network was constructed using Cytoscape 3.7.2. The potential therapeutic targets were imported into the DAVID 6.8 for GO-Biological process （GO-BP） analysis to predict related biological processes which were subsequently verified by the animal experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the effect of HHD on inflammatory factors in colon tissues of mice. Western blot was used to detect the protein expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cysteinyl aspartate-specific protease 3 （Caspase-3）. The IVIS system was used to detect the content of reactive oxygen species （ROS） in colon tissues of mice in each group.ResultNineteen active components of HHD were screened out， involving 32 potential therapeutic targets against UC and 158 biological processes. The results of the animal experiment showed that HHD exerted its anti-UC effect by inhibiting the expression of inflammatory factors tumor necrosis factor （TNF）-α and interleukin-1β （IL-1β）， reducing the content of apoptotic proteins， and regulating the expression of ROS.ConclusionThis study revealed the rationality of predictions and guidance of network pharmacology in experimental design， and confirmed that HHD could exert its effects by participating in biological processes such as immune inflammation， apoptosis， and ROS， which is expected to provide a basis for the mechanism research of HHD in the treatment of UC.Keywords：network pharmacology;ulcerative colitis，inflammation;reactive oxygen species;apoptosis- 36
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Five Views on Differentiation and Treatment-Building New System of Traditional Chinese Medicine Treatment of Tumors
Abstract：As one of the diseases with high incidence in China， cancer seriously endangers human health. The scientific research and clinical practice of traditional Chinese medicine （TCM） in the prevention and treatment of tumors during the Sixth Five-Year Plan period and the 13th Five-Year Plan period show that TCM has certain advantages in preventing and treating postoperative metastasis and recurrence， prolonging survival period， alleviating adverse reactions， and improving the quality of life of patients with advanced tumors. However， innovation of the TCM theoretical thinking and realization of the TCM full-cycle management are needed urgently， which limits the improvement of clinical efficacy. Malignant tumor is a truly representative of major difficult diseases. The simple mode of syndrome differentiation and treatment cannot meet the clinical needs， and thus the triple mode of disease， syndrome and symptom differentiation and treatment emerged， and has received widespread attention. However， since malignant tumors have their own characteristics of occurrence， development and evolvement， it is urgent to establish a new system of TCM differentiation and treatment for special diseases to adapt to the law of modern disease development. Therefore， on the basis of the triple mode， this paper innovatively proposed a new system of cancer prevention and treatment based on five views on differentiation and treatment in TCM， forming a new paradigm of whole-cycle， whole-chain and all-directional integrated Chinese and western medicine prevention and treatment of tumors. Specifically， time-space view： On the basis of the holistic concept and combined with the complex characteristics of different pathological types， lesion location， disease course and treatment stages of malignant tumors， the dynamic and systematic participation of TCM in the whole process of tumor treatment was brought into play from the time and space dimensions. Core view： The core pathogenesis was summarized based on the combination of disease and syndrome， and its key role in guiding differentiation and treatment of malignant tumors was emphasized. Additionally， the pathogenesis characteristics and evolvement rules of various cancer types in different stages were paid attention. Symptom view： The symptoms were ameliorated and the quality of life was improved. The current obvious contradictions of patients were solved to enhance the humanistic nature of treatment. Precision view： In combination of modern medical concepts， TCM constitutions and laboratory indicators， TCM advantages were enriched and emphasized for precise clinical positioning. Disease-before-onset view： As prevention is more important than treatment， precaution was focused on in each stage of tumors. The five views had different emphases and were interrelated， covering new understandings of the existing TCM prevention and treatment system of malignant tumors. In addition， new ideas and concepts have been introduced on the basis of the original TCM theory， which provided new strategies for the comprehensive prevention and treatment of tumors.Keywords：five views on differentiation and treatment;traditional Chinese medicine;malignant tumor;comprehensive treatment strategy- 31
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Mahuang Shengmatang Based on Five Depression Syndromes in
Huangdi Neijing Abstract：Mahuang Shengmatang was first recorded in the chapter "Reverting Yin Diseases" in the Treatise on Cold Damage （《伤寒论》）. Whether it is a classic prescription created by ZHANG Zhong-jing is controversial among scholars and physicians due to its distinctiveness in contexts and drug composition. In fact， its proven efficacy in a number of clinical cases cannot be denied， and it is rather biased to judge that Mahuang Shengmatang is a fallacy of later generations. The present study pointed out that Mahuang Shengmatang was a representative in the Treatise on Cold Damage in the treatment of diseases with complicated syndromes. In this formula， although the compatibility of drugs effective in dispersion， astringing， nourishing Yin， assisting Yang， clearing heat， dissipating cold， ascending， floating， sinking， and descending seems complex， it follows certain therapeutic principles. Five depression syndromes in the Huangdi Neijing （《黄帝内经》） highly reflect the Five Elements losing the nature， and embody multiple characteristics of the five elements， such as the movement forms， functions， and disease nature. Furthermore， the corresponding treatment methods were pointed out in the Plain Questions · Major Discussion on the Progress of the Six Climatic Changes of Huangdi′s Internal Classic， including "stagnation of wood should be treated by smoothing， stagnation of fire by dispersing， stagnation of earth by attacking， stagnation of metal by dredging， and stagnation of water by inhibiting"， aiming to restore the physiological conditions of the five elements. The indicated syndromes of Mahuang Shengmatang are complex in superior-inferior， asthenia-sthenia， and cold-heat. The five depression syndromes and their treatment methods recorded in the Huangdi's Internal Classic can reflect the key syndrome pathogenesis and therapeutic principles， which are conducive to analyzing the syndrome differentiation and formulating principles of Mahuang Shengmatang. In the disease progression， impaired Yang Qi in greater Yin and inward invasion of cold pathogens in reverting Yin happen in the initial stage of indicated syndromes of Mahuang Shengmatang， with characteristics of water stagnation， earth stagnation， and wood stagnation. Furthermore， mental stagnation and fire stagnation triggered by Yang Qi accumulation in greater Yang and Yang brightness are the secondary consequences. The compatibility of five types of drugs in Mahuang Shengmatang can act on five depression syndromes in a targeted manner. Based on the five-depression theory in the Huangdi′s Internal Classic and available literature， the present study summarized syndrome pathogenesis and medication characteristics of Mahuang Shengmatang， clarified the correlation between formula and syndrome， and reviewed clinical reports in recent years， which is expected to provide references for the clinical application of Mahuang Shengmatang.Keywords：five depression;Mahuang Shengmatang;classic prescription;ascending， descending， floating， and sinking;reverting Yin disease;exogenous disease- 37
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Classical Famous Prescription Zhenwutang in Treatment of Chronic Heart Failure： A Review
Abstract：Chronic heart failure （CHF）， the end stage of heart disease due to a variety of causes， features high disability rate and mortality， which has become a hot spot in cardiovascular field. As recorded in Treatise on Cold Damage（《伤寒论》）， Zhenwutang is composed of Radix Aconiti Lateralis Preparata， Poria Cocos， Rhizoma Atractylodis Macrocephalae， Paeoniae Radix Alba， and Rhizoma Zingiberis Recens. With the functions of warming Yang and excreting water， it is a classical prescription for the treatment of CHF in clinical settings. By searching China National Knowledge Infrastructure （CNKI）， PubMed， Wanfang Data， and VIP， we find Zhenwutang exerts therapeutic effect on CHF through multiple targets and multiple pathways. Experiments show that it alleviates CHF by antagonizing the overactivation of neuroendocrine system， inhibiting immune-inflammatory response， suppressing cardiac remodeling， restricting apoptosis， regulating autophagy， improving myocardial energy metabolism， inhibiting oxidative stress injury， protecting endothelial function， and decreasing volume load. Clinical research shows that Zhenwutang can significantly alleviate the clinical symptoms of CHF patients in a safe manner with little adverse reactions. This paper systematically summarizes the mechanisms of and clinical research on Zhenwutang in the treatment of CHF in recent years， so as to provide theoretical and experimental data for the further research and development of Zhenwutang.Keywords：Zhenwutang;chronic heart failure;mechanism;clinical applications;research progress- 36
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Multidrug Resistance in Breast Cancer and Coping Strategies of Chinese Medicine： A Review
Abstract：Breast cancer， as a kind of malignant tumor with high incidence rate in female population， poses a great threat to people's health. At present， chemotherapy is the main treatment for breast cancer besides surgery and radiotherapy. However， chemotherapy is often accompanied by multidrug resistance （MDR）， which results in a series of severe consequences such as low efficacy， relapse of cancer after drug withdrawal， increased consumption of medical resources， and increased burden of medical system. Previous studies showed that the mechanism of MDR in breast cancer was quite complex， involving drug efflux， deoxyribonucleic acid （DNA） damage repair， tumor microenvironment， autophagy， epigenetic regulation， tumor stem cells， lipid metabolism， and so on. In addition， there were extensive connections among the mechanisms of MDR. Therefore， it is necessary to reveal the mechanism of MDR and develop corresponding drugs to reverse MDR， thus improving the effect of chemotherapy on treating breast cancer. Chinese medicine has the advantages of high efficiency， low toxicity， multi targets， and overall regulation. Various studies found that Chinese medicine monomer， single drug， and compound were able to reverse MDR in breast cancer by regulating the expression of drug efflux protein， promoting apoptosis， and regulating autophagy， showing the potential of anti-MDR in breast cancer. This paper summarized the research progress of the mechanism of MDR in breast cancer and the strategies of Chinese medicine in coping with MDR in breas cancer in recent years， and provided references for further research.Keywords：breast cancer;multidrug resistance （MDR）;research status;Chinese medicine;coping strategies- 23
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Chinese Medicine in Regulating Signaling Pathways Related to Allergic Rhinitis： A Review
Abstract：Allergic rhinitis （AR）， a common disease in otolaryngology， is intractable with prolonged attack and greatly affects the daily life of patients. Western medicine adopts many therapeutic protocols， such as medication， immunotherapy， and surgery， and also shows disadvantages， including severe side effects and poor long-term curative effect. As reported by modern research， Chinese medicine has the characteristics of good safety， stable curative effect， multi-target and overall regulation， and has unique advantages in the prevention and treatment of AR. With the increasing clinical practice of Chinese medicine in the treatment of AR， scholars have carried out substantial basic research on the regulation of AR signaling pathways by monomers and Chinese medicinal compounds from molecule-cell-biology. To further explain the transduction mechanism of AR signaling pathways， this paper systematically summarized the research progress based on the studies of monomers and Chinese medicinal compounds to provide references for the in-depth research on the intervention of related signaling pathways by Chinese medicine. The conclusions were drawn as follows. The main signaling pathways involved included nuclear factor-kappa B （NF-κB） signaling pathway， TLR signaling pathway， interleukin （IL）-33/growth stimulation expressed gene 2 （ST2） signaling pathway， phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） pathway， and mitogen-activated protein kinase （MAPK） pathway. As revealed， the signaling pathways involved in the treatment of AR by Chinese medicine interacted with each other and genes were not independent in exerting the effects. For example， TLR， as the upstream signal， affected the PI3K/Akt and MAPK signaling pathways， and NF-κB was the downstream substrate of PI3K/Akt， TLR， IL-33/ST2， and MAPK signaling pathways. It was found that IL-33/ST2， as a new signaling pathway， was correlated with the severity and prognosis of AR.Keywords：allergic rhinitis;Chinese medicine;signaling pathway;research progress- 29
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09 -
Harvesting and Primary Processing in
Fritillaria Medicinal Materials： A Review Enhanced PublicationAbstract：Chinese medicinal material from Fritillaria， Beimu in Chinese， is a commonly used antitussive and expectorant traditional Chinese herbal medicine， with the significant functions of clearing heat and moistening lung，resolving phlegm and relieving cough. Five kinds of Fritillaria were recorded in the 2020 edition of Chinese Pharmacopoeia， including Fritillariae Cirrhosae Bulbus， Fritillariae Ussuriensis Bulbus， Fritillariae Thunbergii Bulbus， Fritillariae Hupehensis Bulbus and Fritillariae Pallidiflorae Bulbus. At present， the reports on Fritillaria mainly focus on the pharmacological effect， chemical composition， identification of authenticity and other aspects， while there were few reports on harvesting and primary processing of original medicinal materials. Fritillaria medicines were perennial medicinal plant with various and complex varieties， their quality and curative effect were greatly affected by harvesting and processing in producing area. The processing method differed according to its variety. Fritillariae Cirrhosae Bulbus mainly from western Sichuan plateau， Fritillariae Pallidiflorae Bulbus from Xinjiang and Fritillariae Ussuriensis Bulbus from Northeast China were mostly harvested from June to July and sun dried directly or dried. But Fritillariae Thunbergii Bulbus and Fritillariae Hupehensis Bulbus from Yangtze River basin were harvested when the plants wilted at the beginning of summer， and auxiliary materials such as shell powder and lime must be added during the processing. At present， the drying methods of Fritillaria were still traditional， which is not suitable for large-scale production of cultivated products. Therefore， it is urgent to find a scientific， reasonable and efficient processing methods. Aimed at providing references for standardization production of Fritillaria， this paper made a textual research on the ancient and modern herbal literature， the Chinese Pharmacopoeia and other medicinal standards， combined with modern literature， the harvesting and processing methods of Fritillaria were sorted out and prospected.Keywords：Fritillaria medicinal materials;harvest year;harvest time;producing area processing;drying technology;quality evaluation;herbal textual research- 21
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<HTML><CITATION> <Bulk Citation>Published：2022-06-06 -
Chinese Herbal Prescription Against Esophageal Cancer： A Review
Abstract：Esophageal cancer is a type of upper gastrointestinal malignant tumor with a high degree of malignancy， strong invasion ability， and poor prognosis， which belongs to the category of "dysphagia" in traditional Chinese medicine （TCM）. In addition to tumor resistance， Chinese herbal prescription plays a role in sensitization. In light of information in literature， the syndrome elements of esophageal cancer include Qi stagnation， phlegm， Qi deficiency， blood stasis， and Yin deficiency. After clinical differentiation， the syndromes of esophageal cancer are divided into phlegm and Qi obstruction， Qi and Yin deficiency， fluid deficiency and heat accumulation， Qi deficiency and phlegm dampness， combined phlegm and heat， combined phlegm and stasis， combined heat toxin and stasis， healthy Qi deficiency and toxin accumulation， et al. Dabanxiatang， Qigesan， Xuanfu Daizhetang， Liujunzitang， Shashen Maidongtang， and Tongyoutang are the common clinical prescriptions， where Qigesan， Liu Junzitang， and Tongyoutang have been proved by in vitro and in vivo experiments to exert anti-esophageal cancer effect by directly inhibiting tumor cell proliferation， promoting apoptosis， affecting tumor microenvironment， regulating cell energy metabolism， and inhibiting angiogenesis. In addition， an increasing number of studies have been conducted on anti-esophageal cancer effect of Chinese herbal prescription by targeting non-coding single-stranded microRNA. The specific mechanisms of Da Banxiatang， Shenzhe Peiqitang， Xiao Xianxiongtang， Renshen Banxiatang， and Liushenwan have been scarcely reported despite good clinical efficacy. Wuzhuyu Tang and Tongguansan recorded in ancient books have been rarely applied in modern times. Therefore， the present study reviewed the special drugs and prescriptions mentioned in ancient TCM classics， the commonly used Chinese herbal prescriptions in modern clinical practice， and experimental research progress， to promote treatment methods of Chinese herbal prescriptions against esophageal cancer.Keywords：Chinese herbal prescription;esophageal cancer;Da Banxiatang;Liu Junzitang;Tongyoutang;Xiaoxianxiongtang;Shenzhe Peiqitang;proliferation;apoptosis;microenvironment- 33
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<HTML><CITATION> <Bulk Citation>Published：2022-06-09
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