Abstract：ObjectiveTo explore the effect and mechanism of Xiangshenwan on ulcerative colitis （UC） induced by dextran sulfate sodium （DSS） in mice based on the classic Toll-like receptor （TLR）/nuclear factor kappa B （NF-κB） signaling pathway.MethodThe experimental mice were divided into a normal group， a model group， a Xiangshenwan group， and a mesalazine group. The mice， except for those in the normal group， received 3% DSS solution for 7 days to establish the acute UC model and were treated with Xiangshenwan （5 g·kg-1） and mesalazine （300 mg·kg-1） continuously from the 1st day to the 10th day of modeling. The body weight， disease activity index （DAI）， colon weight， intestinal weight index， colon length， colon weight per unit length， and pathological changes of mice were evaluated respectively. The protein expression of TLR5， myeloid differentiation factor 88 （MyD88）， interleukin-1 receptor-associated kinase 4 （IRAK4）， tumor necrosis factor receptor-associated factor 6 （TRAF6）， transforming growth factor β-activated kinase 1 （TAK1）， p38 mitogen-activated protein kinase （MAPK）， NF-κB， IRAK1， TAK1-binding protein 1 （TAB1）， TAB2， mitogen-activated protein kinase kinase 3 （MKK3）， MKK6 and cyclic adenosine monophosphate response element-binding protein （CREB） in colon tissues of mice was detected by Western blot.ResultCompared with the normal group， the model group showed decreased body weight of mice， increased DAI scores， elevated colon weight， intestinal weight index， and colon weight per unit length， shortened colon length， severe colonic mucosal injury， and up-regulated protein expression of TLR5， MyD88， IRAK4， TRAF6， TAK1， p38 MAPK， NF-κB， IRAK1， TAB1， TAB2， MKK3， MKK6， and CREB in colon tissues （P<0.05， P<0.01）. Compared with the model group， the Xiangshenwan group and the mesalazine group displayed increased body weight of mice， decreased DAI scores， declining colon weight， intestinal weight index， and colon weight per unit length， increased colon length， improved colonic mucosal injury， and down-regulated protein expression of TLR5， MyD88， IRAK4， TRAF6， TAK1， p38 MAPK， NF-κB， IRAK1， TAB1， TAB2， MKK3， MKK6， and CREB in colon tissues （P<0.05， P<0.01）.ConclusionXiangshenwan can effectively treat DSS-induced UC presumedly by the inhibition of TLR/NF-κB signaling pathway.
Abstract：ObjectiveTo investigate the anti-anxious depression mechanism of Baihe Dihuangtang from the NOD-like receptor thermal protein domain 3 （NLRP3） inflammasome.MethodFifty SD rats were randomly divided into normal group， model group， venlafaxine group （13.5 mg·kg-1）， Baihe Dihuangtang high and low dose group （16，4 g·kg-1）， with 10 rats in each group. Chronic restraint stress for 28 days （6 h） combined with subcutaneous injection of corticosterone （30 mg·kg-1） was used to establish induce an anxious depression model. From the 8th day of modeling， the rats in the normal group and the model group received distilled water， and those in groups with drug intervention were treated with corresponding drugs by gavage for 21 days. Elevated plus maze and open field test were used to evaluate the behavioral changes of rats. Enzyme- linked immunosorbent assay （ELISA） was used to detect serum and hippocampal interleukin-1β （IL-1β）， interleukin-6 （IL-6） and interleukin-18 （IL-18） levels. Western blot were used to detect the relative expression of hippocampal NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， and Caspase-1. The pathological changes of the hippocampus were observed by hematoxylin-eosin（HE） staining， the average fluorescence intensity of NLRP3， ASC， and Caspase-1 was detected by immunofluorescence. The ultrastructure of neurons was observed under electron microscopy.ResultCompared with the normal group， the model group showed reduced total entries （TE）， the ratio of open-arm entries （OE%）， the ratio of open-arm times （OT%）， and the autonomous activity score （P<0.01）， significant anxiety and depression-like behaviors， increased levels of IL-1β， IL-6， and IL-18 in the serum and hippocampus （P<0.01）， elevated protein expression of NLRP3， ASC， and Caspase-1 （P<0.01）， activated NLRP3 inflammasomes， and injured hippocampal neurons. Compared with the model group， the high-dose Baihe Dihuangtang group showed improved anxiety and depression-like behaviors （P<0.01）， and decreased levels of IL-1β， IL-6， and IL-18 in the serum and hippocampus （P<0.05，P<0.01）， reduced protein expression of NLRP3， ASC， and Caspase-1 （P<0.01）， and alleviated hippocampal neuron damage.ConclusionBaihe Dihuangtang can improve neuronal damage in anxious depression by inhibiting the excessive activation of NLRP3 inflammasomes.
Keywords：Baihe Dihuangtang;anxiety depression;NOD-like receptor thermal protein domain 3 （NLRP3）;inflammasome;hippocampus;neurons
Abstract：ObjectiveTo explore the underlying protective mechanism of Kaixinsan on learning， memory， and synaptic function in APP/PS1 mice.MethodSixty APP/PS1 mice were randomly divided into a model group， a donepezil （2 mg·kg-1·d-1） group， and low- （0.7 g·kg-1·d-1）， medium- （1.4 g·kg-1·d-1）， and high-dose （2.8 g·kg-1·d-1） Kaixinsan groups， and the wild-type mice of the same age in the same litter were assigned to the normal group， with 12 mice in each group. After continuous intragastric administration for two months， the Morris water maze experiment was performed. The ultrastructure of hippocampal neurons was observed by transmission electron microscopy. The colorimetric assay was used to detect serum content of acetylcholine （ACh）， choline acetyltransferase （ChAT）， acetylcholinesterase （AChE）， and levels of hippocampal reactive oxygen species （ROS）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and glutathione peroxidase （GSH-Px）. Real-time fluorescence-based quantitative polymerase chain reaction （Real- time PCR） was used to detect the mRNA expression of hippocampal brain-derived neurotrophic factor （BDNF）， beta-nerve growth factor （NGFB）， discs large homolog (DLG)2， DLG4， and synaptophysin （SYP）.ResultCompared with the normal group， the model group showed prolonged escape latency， reduced number of crossing platforms， shortened stay in the target quadrant （P<0.01）， decreased number of mitochondria with different shapes and irregular arrangement， some swollen and deformed mitochondria with broken mitochondrial cristae， endolysis， and cytoplasm vacuole， and more cell debris. Additionally， the model group also displayed reduced serum levels of ACh and ChAT， increased AChE （P<0.01）， elevated hippocampal ROS and MDA （P<0.05，P<0.01）， declining SOD and GSH-Px （P<0.01）， and diminished hippocampal BDNF， NGFB， DLG2， DLG4， and SYP mRNA levels （P<0.05，P<0.01）. Compared with the model group， the donepezil group， and the medium- and high-dose Kaixinsan groups showed shortened escape latency， increased number of crossing platforms， prolonged stay in the target quadrant （P<0.05，P<0.01）， improved mitochondrial damage with a regular shape （mainly oval shape）， relieved mitochondrial swelling and deformation， and clear mitochondrial cristae. Furthermore， the donepezil group， and the medium- and high-dose Kaixinsan groups also exhibited increased serum ACh and ChAT levels （P<0.05，P<0.01）， blunted AChE activity （P<0.05）， reduced hippocampal ROS level （P<0.05，P<0.01）， declining MDA level （P<0.05）， potentiated SOD and GSH-Px activities， and up-regulated hippocampal BDNF， NGFB， DLG2， DLG4， and SYP mRNA levels （P<0.05，P<0.01）. In the low-dose Kaixinsan group， the stay time in the target quadrant was prolonged and the expression of hippocampal SYP mRNA was elevated significantly （P<0.05）. There was no statistical difference in swimming speed between the groups.ConclusionKaixinsan can improve the learning and memory ability of APP/PS1 mice by increasing the expression of synaptic plasticity-related proteins， reducing the ultrastructural damage to hippocampal neurons， resisting oxidative stress， and regulating cholinergic neurotransmitters， thereby exerting neuroprotective effects.
Keywords：Kaixinsan;Alzheimer's disease;synaptic plasticity-related proteins;oxidative stress;cholinergic neurotransmitter;learning and memory ability
Abstract：ObjectiveTo investigate the effects of Buyang Huanwutang on the expression of microtubule-associated protein-2（MAP-2）， neurofilament-M（NF-M）， and growth associated protein-43（GAP-43）in rat sciatic nerve after sciatic nerve transection and anastomosis. To explore the mechanism of Buyang Huanwutang promoting peripheral nerve regeneration.MethodSD rats were selected as the experimental subjects， and sciatic nerve transection model was selected as the experimental model. They were randomly divided into model group， sham operation group， Buyang Huanwutang group high， medium and low dose （29.6， 14.8， 7.4 g·kg-1）group， and mecobalamin （0.156 mg·kg-1）group， the model group and the sham operation group were given distilled water intragastric administration. After successful modeling， each group was treated with relevant drugs for 4 weeks. After 4 weeks， sciatic nerve function index（SFI）， degree of inclined plate test and hematoxylin-eosin（HE）of sciatic nerve in each group were tested. The expression levels of MAP-2， NF-M， and GAP-43 at the sciatic nerve anastomosis site were detected by immunohistochemistry and Western blot.ResultCompared with sham operation group， the expression levels of SFI， inclined plate test， MAP-2， NF-M and GAP-43 in model group were significantly increased （P<0.01）. Compared with model group， the expression levels of SFI， inclined plate test， MAP-2， NF-M and GAP-43 in Buyang Huanwutang high， medium and low-dose groups were significantly increased （P<0.05，P<0.01）.ConclusionBuyang Huanwutang has a positive effect on nerve regeneration after sciatic nerve transection and anastomosis in rats.
Abstract：ObjectiveTo explore the mechanism of Banxia Xiexintang （BXXX） in preventing and treating chronic atrophic gastritis （CAG） through Kelch-like ECH-associated protein 1 （Keap1）/nuclear factor erythroid 2-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway.MethodSD rats were divided into a normal group （n=12） and an experimental group for CAG model induction. The model rats were then randomly divided into a model group， a vatacoenayme （VG） group （60 mg·kg-1）， and high- （280 mg·kg-1）， medium- （140 mg·kg-1）， and low-dose （70 mg·kg-1） BXXX groups. The doses in the BXXX groups were equivalent to 28， 14， and 7 g·kg-1 crude drugs. The rats in the normal group and the model group received distilled water at an equal volume， and those in the VG group and the BXXX groups were treated correspondingly by gavage. After 12 weeks of treatment， hematoxylin-eosin （HE） staining was carried out to observe pathological changes in the gastric mucosa of CAG rats. Western blot and real-time fluorescence-based quantitative PCR was used to detect the protein and mRNA expression levels of Nrf2， glutathione S-transferase （GST）， and NAD （P）H：quinone oxidoreductase 1 （NQO1） in the gastric mucosa of CAG rats.ResultCompared with the normal group， the model group showed increased protein and mRNA expression levels of Nrf2， NQO1， and GST in the gastric mucosa of the rats （P<0.05）， atrophic gastric mucosa， and even intestinal metaplasia. The protein and mRNA expression levels of Nrf2， NQO1， and GST in the VG group and the high- and medium-dose BXXX groups were lower than those in the model group （P<0.05）， and gastric mucosa atrophy and intestinal metaplasia were significantly improved， especially in the high-dose BXXX group. However， the effect in the low-dose BXXX group was not significant.ConclusionBXXX can blunt the transcriptional activity of Nrf2， shut down Nrf2 signaling pathway， and reduce the expression levels of NQO1 and GST to achieve normal oxidation-anti-oxidation balance， which may be one of its action mechanisms in the treatment of CAG.
Abstract：ObjectiveTo observe the effects of Wenxin prescription on the key targets of gap 1/synthesis （G1/S） cell cycle transformation in rats with atherosclerosis （AS）， and reveal the mechanism of Wenxin prescription in the treatment of AS.MethodNinety SPF Wistar rats were randomly divided into a normal group （n=6） and a modeling group （n=84）. The rats in the modeling group were fed on a high-fat diet （4% cholesterol， 0.5% sodium cholate， 0.2% propyl thiouracil， 10% lard， 5% sugar， and 80.3% basal feed） for 60 days， and intraperitoneally injected with 400 000 U·kg-1 vitamin D3， once a week for three weeks. The model rats were then randomly divided into a model group， high-dose （24 g·kg-1）， medium-dose （12 g·kg-1）， and low-dose （6 g·kg-1） Wenxin prescription groups， and a rosuvastatin （1.8 mg·kg-1） group. The groups with drug intervention were treated correspondingly by gavage for 30 days. The rats in the model group were administered with an equal volume of distilled water. The general condition of rats was observed after treatment. The levels of high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and total cholesterol （CHO） were detected by enzyme-linked immunosorbent assay （ELISA）， and the atherosclerosis index （AI） was calculated. The pathological morphology of the coronary artery and aorta was observed by hematoxylin-eosin （HE） staining. The protein and mRNA expression of E2F transcription factor 1 （E2F1）， phosphorylated retinoblastoma protein （p-Rb）， cell division cycle 25 （Cdc25）， CyclinE， and CyclinD1 was detected by Western blot and real-time fluorescence-based quantitative polymerase chain reaction （Real-time-PCR）， respectively.ResultCompared with the normal group， the model group showed intima thickening， smooth muscle proliferation， and plaque formation in the coronary artery and aorta， decreased HDL-C （P<0.01）， increased LDL-C， CHO， and AI （P<0.01）， elevated protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE and CyclinD1 （P<0.05）. Compared with the model group， the rosuvastatin group and the Wenxin prescription groups showed slight intimal hyperplasia and lumen narrowing of the coronary artery and aorta， decreased levels of LDL-C， CHO， and AI （P<0.01）， and declining protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE， and CyclinD1 to varying degrees （P<0.05）.ConclusionWenxin prescription can significantly inhibit the expression of key proteins and genes of the G1/S cell cycle， regulate G1/S cell cycle transformation， and reduce vascular smooth muscle and intimal hyperplasia in AS rats.
Abstract：ObjectiveTo compare the effects of Baiyaojian before and after fermentation on intestinal flora and expression of Occludin and zonula occludens protein-1 （ZO-1） in intestinal mucosa of mice with ulcerative colitis （UC）， and to explore the mechanism of Baiyaojian and Galla Chinensis in the treatment of UC.MethodTotally 50 mice were randomly divided into 5 groups with 10 mice in each group， one group was randomly selected as blank group， and the other 4 groups were treated with dextran sodium sulfate （DSS） to induce UC model. After modeling， mice in the blank group and model group were given normal saline， and treatment groups were given Mesalazine （0.8 g·kg-1）， Galla Chinensis decoction （1.8 g·kg-1） and Baiyaojian decoction （2.7 g·kg-1） by intragastric administration for 7 days. The 16S rRNA sequencing technology was used to detect the changes of intestinal flora in mouse feces. The histopathological changes of colon tissue were observed by hematoxylin-eosin （HE） staining， and the expression of Occludin and ZO-1 in colon tissue of mice were compared by immunohistochemistry.ResultCompared with the blank group， the abundance and diversity of intestinal flora in UC mice were significantly decreased， and the colonic tissue was thickened with congestion and obvious ulcers， and the expression levels of Occludin and ZO-1 were significantly decreased （P<0.01）. After treatment with Galla Chinensis and Baiyaojian， the abundance and diversity of flora were improved. At the phylum level， relative abundance of Firmicutes， Proteobacteria and Actinobacteria increased significantly （P<0.01）， while the relative abundance of Bacteroidetes decreased significantly （P<0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of Proteobacteria and Actinobacteria increased significantly （P<0.01）， while the relative abundance of Firmicutes increased and the relative abundance of Bacteroidetes decreased， but there was no significant difference. At the genus level， the relative abundance of Bacteroides， Allobaculum and Ruminococcus decreased significantly （P<0.01）， the relative abundance of Roseburia， Prevotella， Oscillospira and Paraprevotella increased significantly （P<0.05， P<0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of Bacteroides and Allobaculum decreased significantly （P<0.05， P<0.01）， and the relative abundance of Prevotella， Oscillospira， Roseburia and Ruminococcus increased significantly （P<0.01）. Compared with model group， colon tissue of Galla Chinensis group and Baiyaojian group was recovered obviously， congestion was alleviated， only scattered ulcers were seen. The expression of Occludin and ZO-1 increased， and the expression level of Baiyaojian group was higher than that of Galla Chinensis group.ConclusionThe effect of Baiyaojian is better than Galla Chinensis in the treatment of UC. The mechanism may be through regulating the abundance and diversity of intestinal flora， improving the disorder of intestinal flora and increasing the expression of ZO-1 and Occludin and protecting the intestinal mucosal barrier function for alleviating intestinal inflammation.
Abstract：ObjectiveTo investigate the effects of Di'ao Xinxuekang （DXXK） on NLRP3 inflammasome in mouse RAW264.7 macrophages and thoracic aorta of rats with atherosclerosis （AS）， so as to explore its anti-AS mechanism.MethodRAW264.7 cells were stimulated with oxidized low density lipoprotein （ox-LDL） and then intervened with MCC950 and DXXK. The contents of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were determined by enzyme linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of Nod-like receptor protein 3 （NLRP3）， inflammasome adaptor protein apoptosis-associated speck-like protein containing CARD （ASC）， and cysteine-dependent aspartate-directed protease-1 （Caspase-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blotting. Sixty male SD rats were randomly divided into the normal group， model group， atorvastatin group （2.0 mg·kg-1）， as well as high-， medium-， and low-dose （100， 30， and 10 mg·kg-1） DXKK groups， with 10 rats in each group. The rats were exposed to the high-fat diet and vitamin D2 for inducing AS. The blood lipid level was measured using an automatic biochemical analyzer， followed by the calculation of AS index （AI）. The contents of serum TNF-α and IL-1β were determined by ELISA， and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in thoracic aorta were assayed by Real-time PCR and Western blotting. HE staining and Sirius red staining were conducted to observe the pathomorphological changes in the abdominal aorta and aortic sinus.ResultCompared with the normal group， the model group exhibited significantly increased TNF-α and IL-1β contents and up-regulated NLRP3， ASC， and Caspase-1 mRNA and protein expression in RAW264.7 cells （P<0.01）. The above indexes in each drug administration group were significantly reduced in contrast to those in the model group （P<0.05， P<0.01）. The comparison with the model group showed that cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and AI in each DXXK group significantly declined， while the high-density lipoprotein cholesterol （HDL-C） was significantly elevated （P<0.05， P<0.01）. The levels of serum TNF-α and IL-1β and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in the thoracic aorta were decreased （P<0.05， P<0.01）. Abdominal aortic lesions and fibrous hyperplasia of aortic sinus were significantly improved.ConclusionDXXK has a significant anti-AS effect， which is possibly related to the inhibition of NLRP3 inflammasome.
Keywords：Di'ao Xinxuekang （DXXK）;NLRP3 inflammasome;atherosclerosis （AS）;mechanism of action
Abstract：ObjectiveTo investigate the mechanism of Duanteng Yimu decoction （DTYM） in the inhibition of pannus formation in collagen-induced arthritis （CIA） mice.MethodTwenty-four SPF-grade DBA/1 male mice were randomly divided into the following four groups： a blank group （NC group）， a model group （CIA group）， a methotrexate group （MTX group）， and a DTYM group， with six mice in each group. The mice， except for those in the NC group， were modeled. From the second immunization， the medium， MTX （1 mg·kg-1）， and DTYM （15.4 g·kg-1） were administered at an equal volume by gavage for 35 days. Mice were observed for general condition and the arthritis index. The knee and ankle joints were scanned by microcomputed tomography （micro CT）. Hematoxylin-eosin （HE） and safranin O/fast green staining were performed to observe pathological changes. Immunohistochemistry was performed to detect the expression of platelet/endothelial cell adhesion molecule-1 （CD31）， vascular endothelial growth factor-α （VEGF-α）， vascular endothelial growth factor receptor 2 （VEGFR2）， and phosphorylated（p）-VEGFR2.ResultCompared with the NC group， the CIA group showed red and swollen ankle joints， increased arthritis index scores （P<0.05， P<0.01）， manifest injury in the knee and ankle joints， reduced cartilage thickness， elevated Micro CT bone destruction scores of knee and ankle joints （P<0.01）， and up-regulated absorbance values of synovial CD31， VEGF-α， VEGFR2， and p-VEGFR2 （P<0.01）. Compared with the CIA group， the DTYM group showed relieved ankle joint redness and swelling， reduced arthritis index scores of mice three weeks after administration （P<0.05， P<0.01）， intact joint surfaces of the knee and ankle joints， thickened cartilage， declining Micro CT bone destruction scores in both the knee and ankle joints （P<0.05， P<0.01）， and lowered absorbance values of CD31， VEGF-α， VEGFR2， and p-VEGFR2 in the synovium （P<0.01）.ConclusionDTYM can inhibit the pannus formation in CIA mice presumedly by regulating the VEGF pathway.
Abstract：ObjectiveTo explore the effect of Shugan Yishen prescription（SGYS） on the tamoxifen （TAM） -resistant cell line LCC9 by the intervention of exosome-mediated crosstalk in the breast cancer microenvironment.MethodFour groups of serum were set up， specifically， a blank group， a TAM （2 mg·kg-1·d-1） group，an SGYS（113.2 g·kg-1·d-1） group，and a combination group. The exosomes of LCC9 cells were extracted by ultracentrifugation and identified by transmission electron microscopy （TEM），nanoparticle tracking analysis （NTA）， and Western blot. Then the collected LCC9 exosomes （LCC9-EXO） were co-cultured with bone marrow mesenchymal stem cells（BMMSCs），and 10% of the above four groups of serum were added to the co-culture system. After 48 hours of co-culture，the exosomes of BMMSCs （BMMSCs-EXO） were extracted and incubated with LCC9 cells. Fluorescence microscope was used to observe the uptake of exosomes by cells. Cell counting kit-8 （CCK-8） assay，flow cytometry， and Transwell assay were used to detect the effects of drug-containing serum in the four groups on the proliferation，apoptosis， and migration of LCC9 cells. Western blot was used to detect the protein expression of CD24，CD44，human epidermal growth factor 2（HER2）， and estrogen receptor α （ERα） in each group.ResultFluorescence microscope observed that LCC9-EXO could be taken up by BMMSCs，and BMMSCs-EXO could be taken up by LCC9 cells. CCK-8 assay revealed that compared with the TAM group，the SGYS group and the combination group showed reduced cell proliferation ability at each period （P<0.05），especially the combination group，but no statistically significant difference between the SGYS group and the combination group was observed （P<0.05）. Flow cytometry revealed that compared with the TAM group，the SGYS group and the combination group showed increased levels of apoptosis （P<0.05）. Transwell assay revealed that compared with the TAM group，the SGYS group and the combination group showed decreased cell migration ability （P<0.05）. Western blot revealed that compared with the TAM group，the SGYS group and the combination group showed up-regulated expression of ERα and CD24（P<0.05），and down-regulated expression of HER2 and CD44 （P<0.05）. The effect of the combination group on protein expression was superior to that of the SGYS group （P<0.05）.ConclusionSGYS reverses the TAM resistance of LCC9 cells by interfering with the crosstalk between BMMSCs-EXO and LCC9-EXO.
Abstract：ObjectiveTo study the protective effect of the Wenyang Huoxue Huatan prescription （WYHXHT） on cardiotoxicity induced by adriamycin.MethodSD rats were randomly divided into the following six groups： a normal control group， an adriamycin model group， a low-dose （4.86 g·kg-1） WYHXHT group， a middle-dose （9.72 g·kg-1） WYHXHT group， a high-dose （19.44 g·kg-1） WYHXHT group， and a dexrazoxane group. Except for the normal control group， the rats in other groups received intraperitoneal injection of 2.5 mg·kg-1 adriamycin， once a week for six weeks， with a cumulative dose of 15 mg·kg-1. The normal control group， the adriamycin model group， and the dexrazoxane group received 10 mL·kg-1 normal saline daily by gavage. In the dexrazoxane group， the rats were subjected to intraperitoneal injection of 25 mg·kg-1 dexrazoxane 30 min before doxorubicin administration， once a week for six weeks. The general condition of rats was observed and their body weight was monitored. A high-resolution micro-ultrasound imaging system was used to detect rat cardiac function. Hematoxylin-eosin （HE） staining was performed to observe the pathological changes of myocardial tissues of rats. Western blot was used to detect the protein expression of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ， the mammalian homolog of yeast Atg6 （Beclin-1）， and p62 protein in rat myocardial tissues.ResultCompared with the normal control group， rats in the adriamycin model group showed dull fur， reduced food intake and activity， loose stool， low energy， and slow response. Besides， it also displayed reduced body weight （P<0.01）， decreased left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） （P<0.01）， myocardial cell degeneration， edema， rupture， and dissolution， expansion of myocardial interstitium， uneven staining of myocardial fiber， visible inflammatory cell infiltration， up-regulated expression of Beclin-1 and LC3Ⅱ in rat myocardial tissues （P<0.01）， and down-regulated p62 expression （P<0.01）. Compared with the adriamycin model group， the medium- and high-dose WYHXHT groups exhibited increased body weight， LVEF， and LVFS （P<0.01）， relieved pathological injury of myocardial tissues， down-regulated expression of LC3Ⅱ and Beclin-1 （P<0.05，P<0.01）， and up-regulated expression of p62 （P<0.05，P<0.01）.ConclusionWYHXHT can effectively prevent and treat adriamycin-induced cardiotoxicity， and its effect may be related to the inhibition of myocardial cell autophagy. The effect is dominant in the high-dose group.
Keywords：blood activating and phlegm resolving;adriamycin;autophagy;cardiotoxicity
Abstract：ObjectiveTo observe the effect of Jinxiangdan （JXD） on NOD-like receptor pyrin domain-containing-3 （NLRP3）/cysteine-dependent aspartate-directed protease-1 （Caspase-1）/interleukin-1β （IL-1β） signaling pathway in myocardium of rats with myocardial ischemia-reperfusion injury （MIRI） and explore the protective effect and mechanism of JXD against MIRI.MethodFifty male SD rats were randomly divided into the sham operation group， model group， high- and low-dose JXD groups， and positive drug （Di'ao Xinxuekang） group， with 10 rats in each group. Seven days before modeling， rats in the JXD groups were separately treated with intragastric administration of 0.72 and 0.18 g·kg-1 JXD tablets， the ones in the sham operation group and model group with the same volume of normal saline， and those in the positive drug group with 1.29 g·kg-1 Di'ao Xinxuekang. Twelve hours after the last intragastric administration， the anterior descending branch of the left coronary artery was ligated for 30 min and then re-perfused for 60 min for inducing MIRI. ST segment elevation was detected by electrocardiogram（ECG） for model evaluation. The contents of creatine kinase （CK） and lactate dehydrogenase （LDH） in cardiac tissue were measured by colorimetry. Hematoxylin-eosin （HE） staining was conducted for observing myocardial histopathological changes， followed by the detection of cardiomyocyte apoptosis by DNA in situ nick end-labeling （TUNEL） assay. The protein and mRNA expression levels of NLRP3， Caspase-1， and IL-1β were detected by Western blot and real-time polymerase chain reaction （Real-time PCR）， respectively.ResultCompared with sham operation group， the model group exhibited obviously elevated ST segment （P<0.01）， enhanced CK and LDH activities in the myocardium （P<0.01）， increased apoptotic cardiomyocytes （P<0.01）， and up-regulated NLRP3， Caspase-1， and IL-1β protein and mRNA expression （P<0.01）. Compared with model group， JXD at both the high and low doses and Di'ao Xinxuekang significantly lowered the ST segment （P<0.05，P<0.01）， diminished the CK and LDH activities in myocardial tissue （P<0.05，P<0.01）， improved the apoptosis of cardiomyocytes （P<0.05，P<0.01）， and down-regulated the mRNA and protein expression levels of NLRP3， Caspase-1， and IL-1β in myocardial tissue （P<0.05，P<0.01）. The ST segment of ECG in the low-dose JXD group was increased as compared with that in the Di'ao Xinxuekang group （P<0.05）， while the ST segment in the high-dose JXD group was obviously elevated （P<0.05）. Besides， the green fluorescence intensities in the low- and high-dose JXD groups and the Di'ao Xinxuekang group remarkably declined （P<0.05，P<0.01）. The mRNA and protein expression levels of NLRP3， Caspase-1， and IL-1β in the high-dose JXD group were down-regulated （P<0.05）.ConclusionJXD alleviates MIRI possibly by lowering NLRP3 and IL-1β expression and inhibiting cardiomyocyte apoptosis.
Abstract：ObjectiveTo investigate the effect of licochalcone A （LCA） on apoptosis in human breast cancer MDA-MB-231 cells， and to explore its possible mechanism.MethodMDA-MB-231 cells were treated with LCA of different concentrations， and cell counting kit-8 （CCK-8） assay was used to detect the cell viability. The cells were treated with LCA （10， 20， and 40 μmol·L-1） for 24 h， and apoptosis was detected by Annexin V staining with fluorescein isothiocyanate （FITC） and propidium iodide （PI） （Annexin V-FITC/PI）. The level of intracellular reactive oxygen species （ROS） was detected by 2′，7′-dichlorodihydrofluorescein diacetate （DCFA-DA） fluorescent probe. Mitochondrial membrane potential （MMP） was detected by 5， 5′， 6， 6′-tetrachloro-1， 1′， 3， 3′-tetraethyl-imidacarbocyanine （JC-1） fluorescence probe. Western blot was used to detect the expression of cell apoptosis-related proteins， such as B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）， and endoplasmic reticulum （ER） stress-related proteins， such as C/EBP homologous protein （CHOP）， activating transcription factor 4 （ATF4）， protein kinase R-like ER kinase （PERK）， p-PERK， eukaryotic translation initiation factor 2 alpha （eIF2α）， and p-eIF2α.ResultWith the increase in the drug concentration （starting from 5 μmol·L-1）， the cell viability decreased （P<0.05） with IC50 of 19.05 μmol·L-1 as compared with the normal group. Additionally， the apoptosis rates of the LCA groups （10， 20， 40 μmol·L-1） significantly increased （P<0.05）， which reached 30.2% （P<0.05） at LCA concentration of 40 μmol·L-1. LCA （10， 20， and 40 μmol·L-1） decreased the expression of Bcl-2 （P<0.05） and increased Bax expression （P<0.05） in a dose-dependent manner. Besides， the intracellular ROS level was elevated （P<0.05） and mitochondrial MMP was reduced （P<0.05） after LCA （10， 20， and 40 μmol·L-1） treatment in a dose-dependent manner， leading to mitochondrial dysfunction. LCA （10， 20， and 40 μmol·L-1） induced ER stress to up-regulate the expression of CHOP， ATF4， p-PERK， and p-eIF2α （P<0.05） in a dose-dependent manner.ConclusionLCA can induce MDA-MB-231 cell apoptosis by increasing intracellular ROS level and reducing MMP to trigger mitochondrial dysfunction and ER stress.
Keywords：licochalcone A （LCA）;breast cancer;reactive oxygen species;mitochondrial membrane potential;endoplasmic reticulum stress
Abstract：ObjectiveTo study the effect of icaritin on the proliferation， apoptosis， migration and invasion of human epithelial ovarian cancer A2780 cells and the inhibitory mechanism of icaritin against cell invasion and migration via the regulation of epithelial-mesenchymal transformation （EMT）-related molecule expression.MethodA2780 cells were divided into the blank control group and low-， medium-， and high-dose （5， 10， 20 μmol·L-1） icaritin groups and received the corresponding inventions for 48 h. Cell proliferation and viability were detected using the cell counting kit-8 （CCK-8）. The cellular proliferation inhibition and apoptosis rates were assayed by flow cytometry. The cell invasion and migration were observed in Scratch test and transwell test， followed by the calculation of wound healing rate and migration rate. The protein and mRNA expression levels of EMT-related molecules including E-cadherin， N-cadherin， and Vimentin and tumor invasion and migration-related molecule matrix metalloproteinase-9 （MMP-9） were measured by Western blot and real-time polymerase chain reaction （Real-time PCR）.ResultAs revealed by CCK-8 assay and flow cytometry， compared with the blank control group， the icaritin groups all exhibited elevated proliferation inhibition rate （P<0.01） and apoptosis rate （P<0.05）. According to the Scratch test and transwell test， compared with the blank control group， the icaritin groups displayed weakened invasion and migration ability and decreased number and rate of cell invasion and migration （P<0.05）. Western blot and Real-time PCR results showed that the protein and mRNA expression levels of N-cadherin， MMP-9 and Vimentin in each icaritin group were down-regulated as compared with those in the blank control group， while the expression of E-cadherin was up-regulated.ConclusionIcaritin inhibits the proliferation and promotes the apoptosis of human ovarian cancer A2780 cells， and it inhibits the invasion and migration of A2780 cells possibly by regulating the expression of EMT-related molecules.
Keywords：icaritin;ovarian cancer A2780 cells;epithelial-mesenchymal transformation （EMT）;invasion and migration;proliferation and apoptosis
Abstract：ObjectiveTo explore the clinical effects of Xingshen Tongqiao Huoxue decoction in the treatment of dysphagia caused by cricopharyngeal achalasia （CPA） after cerebral infarction and its influence on neurotransmitters.MethodOne hundred and fourteen eligible patients were randomly divided into a control group （56 cases） and a treatment group （58 cases）. Patients in both groups received balloon catheter dilatation and rehabilitation training. In the observation group， the Tiandan Tongluo capsules were further orally taken， five capsales per time， three times per day. In the control group， the Xingshen Tongqiao Huoxue decoction was provided， one dose per day. The treatment lasted for four weeks. Following the videofluoroscopic swallow study （VFSS） and water-swallowing test （WST） before and after treatment， the swallowing quality of life questionnaire （SWAL-QOL）， functional oral intake scale （FOIS） and National Institutes of Health Stroke Scale （NIHSS） scores were calculated. The serum brain-derived neurotrophic factor （BDNF）， substance P （SP）， dopamine （DA）， 5-hydroxytryptamine （5-HT） and nerve growth factor （NGF） levels before and after treatment were detected， and the complications were recorded.ResultThe clinical efficacy of the treatment group was superior to that of the control group （Z=2.079， P<0.05）. Both VFSS and WST findings in the treatment group were milder than those in the control group （Z=2.004， P<0.05 and Z=1.973， P<0.05）. The VFSS， SWAL-QOL， and FOIS scores and the BDNF， DA， SP， 5-HT and NGF levels of the treatment group were elevated in contrast to those of the control group （P<0.01）， whereas the NIHSS score declined （P<0.01）. The complication rate in the treatment group was （8/58） 13.79%， significantly lower than （17/56） 30.36% in the control group （χ2=4.565， P<0.05）.ConclusionOn the basis of balloon catheter dilatation and rehabilitation training， the Xingshen Tongqiao Huoxue decoction can significantly enhance the swallowing function， improve the quality of life， and reduce complications in patients with dysphagia caused by CPA after cerebral stroke.
Abstract：ObjectiveTo construct the targeting evaluation method of traditional Chinese medicine （TCM） preparations based on supramolecular Qi chromatography theory， and to study the liver targeting effect of Bupleuri Radix on Pien Tze Huang.MethodThe molecular connectivity index （MCI） was used to analyze the characteristics of imprinted template and liver targeting tendency of TCM mainly attributed to liver meridian and components of Pien Tze Huang， and combined with target dynamics and total statistical moment principle， aimed at the independent action characteristics of multi-component imprinted template of TCM， a method for evaluating the targeting of TCM preparations was established. Hepatoma rats in Pien Tze Huang group， Bupleuri Radix group， Pien Tze Huang+Bupleuri Radix group and blank group were tested and verified.ResultAfter the average value of MCI of TCM mainly attributed to liver meridian was deducted， the MCI similarity between Pien Tze Huang group and Bupleuri Radix group was 0.376 8， Pien Tze Huang+Bupleuri Radix group and Bupleuri Radix group was 0.988 2， so it was predicted that Bupleuri Radix could enhance the liver targeting of Pien Tze Huang. A system for evaluating the targeting of TCM compounds was established， including relative total uptake efficiency （RUET）， relative total concentration （RCT）， relative imprinted tendency （RITT） and relative imprinted variance （RIVT）. The RUET and RCT of liver were the highest in all tissues （RUET=1.88>1，RCT=2.30>1）， and the corresponding values of other tissues were all <1， indicating that Pien Tze Huang combined with Bupleuri Radix could increase its distribution in liver and enhance liver targeting. Except for plasma， the RITT and RIVT of other tissues fluctuated around 1.0， indicating that targeted modification did not change imprinted tendency of Pien Tze Huang and had no significant effect on the types of components.ConclusionUnder the guidance of supramolecular Qi chromatography theory， a targeting evaluation parameter system can be established to characterize the multi-component imprinted effect of TCM preparations by MCI and total statistical moment parameters， so as to realize the evaluation of targeting of TCM preparations. The addition of Bupleuri Radix can increase the liver targeting of Pien Tze Huang.
Abstract：ObjectiveTo investigate the physiological response of Ginseng Radix et Rhizoma with different phenotypes to high light stress under farmland cultivation mode， and to provide theoretical support for breeding of and fine management of new varieties of farmland ginseng.MethodFour-year-old ginseng plants with different stem colors were used as materials， and blue film was replaced by the colorless and translucent film for performing high light stress and investigating the change of growth status， light response curve， photosynthetic and fluorescence parameters of ginseng leaves. Meanwhile， the physiological adaptability and cell ultrastructural differences of ginseng with different phenotypes for strong light were compared by analyzing the variation of leaf ultrastructural characteristics before and after the stress.ResultThe color of ginseng leaves became weak and the content of chlorophyll was greatly decreased with burning phenomenon on the leaf margin under high light stress. The maximum net photosynthetic rate and light saturation point of purple-stem ginseng were higher than those of green-stem ginseng. Transpiration rate， water use efficiency and other gas exchange parameters of purple-stem ginseng had adversity advantages， photosynthetic electron transport rate， maximum photochemical efficiency， photochemical quenching coefficient and other chlorophyll fluorescence parameters were higher than those of green-stem ginseng. There were changes in ultrastructure of the two germplasms， and the plasmolysis of green-stem ginseng was more obvious， its chloroplast membrane ruptured， the contents were exuded and the accumulation of starch grains and osmiophilic bodies increased. The chloroplast membrane structure of purple-stem germplasm was relatively stable， and its number of multivesicular bodies increased significantly by comparing with that of green-stem ginseng under high light.ConclusionHigh light stress inhibits the leaf growth of farmland ginseng with different stem colors. The photosynthetic efficiency of green-stem ginseng is obviously reduced， and the integrity of chloroplast is damaged， leading to the weak physiological resistance. The purple-stem ginseng can reduce the damage of the photosynthetic system of plants under high light stress by its special cell structure and non-photochemical quenching advantages. Therefore， the purple-stem ginseng can be cultivated into a new resistant variety.
Keywords：Ginseng Radix et Rhizoma;farmland planting ginseng;chlorophyll fluorescence;ultrastructure;photosynthesis;new varieties of stress resistance;light stress
Abstract：ObjectiveTo identify the anti-acetylcholinesterase active ingredients in Aconitum tanguticum， so as to lay the foundation for finding new anti-Alzheimer's disease （AD） drugs.MethodThe anti-acetylcholinesterase active fractions of A. tanguticum were screened by the modified Ellman's method， and the chemical composition of the active fraction was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The chromatographic separation was performed on an ACQUITY UPLC BEH C18 column （2.1 mm×50 mm， 1.7 μm） with acetonitrile （A）-0.4% ammonia aqueous solution （B） as mobile phase for gradient elution， and the column temperature was set at 30 ℃ with the flow rate of 0.4 mL·min-1. Phase A of the dichloromethane fraction changed with time as follows：0-3 min， 5%A； 3-7 min， 5%-20%A； 7-11.5 min， 20%-33%A； 11.5-15.5 min， 33%-50%A； 15.5-20.5 min， 50%-80%A； 20.5-23 min， 80%-85%A； 23-25 min， 85%-95%A. Phase A of the n-butanol fraction changed with time as follows：0-2 min， 5%A； 2-8 min， 5%-20%A； 8-11 min， 20%-33%A； 11-15 min， 33%-95%A. Mass spectrometry was performed on electrospray ionization， data were collected in positive ion mode， and the detection range was m/z 100-1 500.ResultBoth the dichloromethane and n-butanol fractions had a certain inhibitory effect on acetylcholinesterase， their half inhibitory concentration （IC50） values were （64±4.4） mg·L-1 and （85.7±3.8） mg·L-1， respectively. By UPLC-Q-TOF-MS/MS analysis， a total of 21 alkaloids were identified from the dichloromethane fraction， and 11 alkaloids were identified from n-butanol fraction. Guan-fu base Ⅰ， found in both fractions， was first discovered in A. tanguticum.ConclusionDiterpene alkaloids are the main anti-acetylcholinesterase substances of A. tanguticum， which is worth further exploration.
Keywords：Aconitum tanguticum （Bangge）;acetylcholinesterase;ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）;diterpene alkaloids;guan-fu base Ⅰ;dichloromethane fraction;n-butanol fraction
Abstract：ObjectiveTo explore the effect of natural decay of exogenously added fibrous roots on the growth and development of Paris polyphylla var. yunnanensis and its medicinal quality.MethodThe effects of natural decay of fibrous roots at different amounts on mycorrhizal infection rate， physiological and biochemical indexes， and saponin contents of P. polyphylla var. yunnanensis were studied in pot culture experiments at room temperature.ResultThe results showed that the infection rate of arbuscular mycorrhizal （AM） fungi in the root of P. polyphylla var. yunnanensis was not significantly affected by different fibrous root treatments， but there were significant differences in infection intensity. The photosynthetic pigment content in the leaves declined significantly with the increase in fibrous root amount， and the total chlorophyll was decreased by 78.7% at most. The contents of soluble protein， soluble sugar and malondialdehyde in the leaves of P. polyphylla var. yunnanensis showed an overall upward trend. The activities of the three protective enzymes varied. The peroxidase and malondialdehyde were reduced by 181.6% and 200.0% at most. In the root system of P. polyphylla var. yunnanensis， the contents of the above-mentioned six components decreased to varying degrees， with the largest reductions of peroxidase and malondialdehyde reaching 44.6% and 69.7%. Different fibrous root treatments resulted in a decrease in active component content of P. polyphylla var. yunnanensis. The total content of the four saponins was decreased by 58.9% at most， and the total saponin content by 46.9%.ConclusionThe natural decay of fibrous roots affects the soil microbial environment of root system， reduces the photosynthetic pigment content in leaves， and destroys the stability of cells， thus interfering with the growth and development of P. polyphylla var. yunnanensis， reducing its medicinal components， and causing continuous cropping obstacles.
Keywords：Paris polyphylla var. yunnanensis;fibrous root decay;continuous cropping obstacle;infection rate;physiological and biochemical;active components
Abstract：ObjectiveTo establish a method for the determination of artemisinin and arteannuin B in different Artemisia annua germplasms， compare the differences of the two compounds among different A. annua germplasm under the condition of hydroponic homogenization and explore the key factors affecting contents of principal compounds in different A. annua germplasms.MethodSeedlings from different A. annua germplasms were arranged randomly and fed in a hydroponic cultivation system. Contents of artemisinin and arteannuin B were detected by ultra performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS） with multi-reaction monitoring mode and ACQUITY UPLC® BEH C18 column （2.1 mm×100 mm， 1.7 μm）， mobile phase was water-acetonitrile （95∶5， containing 0.1% formic acid， A） and acetonitrile-water （95∶5， containing 0.1% formic acid， B） for gradient elution （0-3.5 min， 25%-1%A； 3.5-3.6 min， 1%-25%A； 3.6-5.0 min， 25%A）， the flow rate was set at 0.4 mL·min-1. The content differences of artemisinin and arteannuin B in different provenances of A. annua were detected and analyzed statistically.ResultThe established method had high sensitivity and good separation. A significant difference of artemisinin and arteannuin B contents was observed in different germplasms under the same culture conditions， that is， under the constant temperature of 25 ℃ in hydroponics. The provenance with higher artemisinin content was Yunnan， and the content was 3 810.597 μg·g-1. The highest strain of arteannuin B was Shanxi provenance germplasm with the content of 1 691.747 μg·g-1. According to the content of artemisinin， the provenances were arranged as follows：Yunnan， Hainan， Hubei， Guangxi， Zhejiang， Shanxi， Beijing， Shandong， Heilongjiang， and Gansu province germplasms. Correlation analysis showed that there was a significant negative correlation between artemisinin content and latitude of A. annua， but there was no significant correlation between contents of artemisinin and arteannuin B and longitude.ConclusionThe contents of artemisinin and arteannuin B among different A. annua germplasms were significantly different under the same culture environment， and the dominant factors affecting biosynthesis and accumulation of artemisinin and arteannuin B in A. annua may be the genetic background， suggesting that germplasm improvement is the key factor to improve the medicinal quality of A. annua in subsequent cultivation.
Keywords：Artemisia annua;artemisinin;arteannuin B;hydroponics;germplasm;ultra performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS）;longitude and latitude
Abstract：ObjectiveTo explore the potential mechanism of Bianketong tablet （BKT） in the treatment of constipation-predominant irritable bowel syndrome （C-IBS） based on network pharmacology and bioinformatics.MethodThe BKT-meridian network was constructed for analyzing the combined effect of the nine Chinese herbs in BKT. The active components and targets of BKT were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and then screened according to the oral bioavailability （OB） and drug likeness （DL） criteria. Following the acquisition of C-IBS target set from GeneCards， Online Mendelian Inheritance in Man （OMIM）， Drugbank and DisGeNet， the protein-protein interaction （PPI） network was constructed. Cytoscape 3.7.2 was utilized for network visualization. The screened key targets were subjected to gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using DAVID platform. The C-IBS mouse model was established via intragastric administration of ice water， and the key targets of BKT against C-IBS were verified by enzyme linked immunosorbent assay （ELISA） and immunohistochemistry.ResultThe large intestinal meridian was the main site where BKT acted. There were 70 potential active components in BKT， which acted on 227 intersection targets. Through T helper cell 17（Th17） differentiation， Toll-like receptor （TLR）， tumor necrosis factor and other signaling pathways， BKT participated in inflammatory response， immune regulation， intestinal nerve regulation， hormonal regulation， and oxidative stress response， thus exerting the therapeutic effects against C-IBS. As reveled by in vivo experiments， BKT significantly improved the small intestinal propulsion rate， up-regulated the expression of vasoactive intestinal peptide （VIP） in serum and colon tissue of C-IBS mice， and down-regulated the expression of nuclear transcription factor-κB （NF-κB）， interleukin（IL）-6， and TLR2 in serum and colon tissue， which confirmed the reliability of integration analysis.ConclusionBKT inhibits C-IBS via multiple components， multiple targets， and multiple pathways. This study has provided ideas for further clinical research and experimental verification of BKT in the treatment of C-IBS.
Abstract：ObjectiveTo explore the key gens of peripheral blood mononuclear cells in hepatocellular carcinoma（HCC-PBMC） and potentially effective Chinese herbs based on bioinformatics， and to verify the clinical efficacy of these Chinese herbs via a systematic review.MethodThe chips GSE58208 and GSE36076 of HCC-PBMC were downloaded from the Gene Expression Omnibus （GEO）， followed by the identification of differentially expressed genes （DEGs） using RStudio. After protein-protein interaction （PPI） analysis by STRING， the DAVID was employed for gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis. The DEGs of HCC-PBMC were visualized by Cytoscape. The key genes of HCC-PBMC were calculated by CytoHubba plug-in and mapped with those in Coremine Medical for screening out the potential Chinese herbs for the treatment of HCC， which were then included for subsequent systematic review.ResultA total of 203 DEGs were obtained （194 up-regulated and nine down-regulated）. As revealed by DAVID analysis， the DEGs were mainly enriched in such biological processes and signaling pathways as transcriptional regulation of DNA template， hydrolysis of ribonucleic acid phosphodiester bond， positive regulation of intranuclear mitosis and division， skeletal muscle fiber development， activation of mitogen-activated protein kinase（MAPK）activity， Fanconi anemia pathway， and metabolic pathway. The key genes of HCC-PBMC were calculated by Cytoscape to be GTPase IMAP family member 1 （GIMAP1）， GTPase IMAP family member 4 （GIMAP4）， GTPase IMAP family member 6 （GIMAP6）， GTPase IMAP family member 7 （GIMAP7）， GTPase IMAP family member 8 （GIMAP8）， interleukin-1β （IL-1β）， CX3C chemokine receptor 1 （CX3CR1）， C-C chemokine receptor type 2 （CCR2）， Toll-like receptor 7（TLR7）， and epidermal growth factor（EGF）. Through Coremine Medical analysis， it was concluded that Ginseng Radix et Rhizoma， Curcumae Longae Rhizoma， Centellae Herba， and Hedyotidis Herba were closely related to the key genes. Ginseng Radix et Rhizoma has the effects of tonifying and benefiting lung and spleen and enhancing strength， suitable for the liver depression and spleen deficiency syndrome or Qi-Yin deficiency syndrome of HCC. Hence， Si Junzitang with Ginseng Radix et Rhizoma as the sovereign medicinal was included for systematic review. It has been confirmed that Ginseng Radix Et Rhizoma was superior to western medicine alone in improving the overall clinical efficacy， alleviating TCM syndrome， elevating serum CD4+ and CD4+/CD8+ levels， and reducing the serum CD8+ and TBIL levels （P<0.01）， with high safety.ConclusionThis study conducted at the gene level has provided new ideas for clinical diagnosis and treatment of HCC. The systematic review of Ginseng Radix Et Rhizoma against HCC provides a basis for the clinical prevention and treatment of HCC with TCM.
Keywords：peripheral blood mononuclear cells in hepatocellular carcinoma（HCC-PBMC）;key genes;traditional Chinese medicine;bioinformatics analysis;systematic review
Abstract：ObjectiveAs the problem of global aging intensifies，postmenopausal osteoporosis （PMOP） has become a global health problem among females. At present，the commonly used biological agents have been proved not suitable for long-term use due to multiple adverse reactions. Several Meta-analyses have confirmed the good safety and effectiveness of kidney-tonifying method against PMOP，but its therapeutic mechanism remains unclear. The purpose of this Meta-analysis was to evaluate the effect of kidney-tonifying method on osteoclastogenesis inhibitory factor（OPG）/receptor activator of nuclear transcription factor （NF）-κB （RANK）/receptor activator of NF-κB ligand （RANKL） signaling pathway in PMOP animal model，so as to provide an experimental basis for the treatment of PMOP with kidney-tonifying method.MethodThe related articles were retrieved from PubMed，Ovid Medline，Embase，China National Knowledge Infrastructure （CNKI），Chongqing Weipu Database for Chinese Technical Periodicals （VIP），and Wanfang Data Knowledge Service Platform with the retrieval time set from their inception to January 2020. The quality of each included article was evaluated using the SYRCLE's risk of bias tool. Then RevMan 5.3 was utilized for Meta-analysis according to the Cochrane systematic review methodology.ResultThirty-two studies involving 619 rats were included. The quality score of these studies ranged from 3 to 5 points. The results of the Meta-analysis indicated obvious advantages of kidney-tonifying method in increasing bone mineral density （BMD）［standardized mean difference （SMD）=2.01，95% confidence interval（CI）=1.50-2.52，P<0.000 01］），serum OPG level （SMD=3.33，95% CI=2.59-4.07，P<0.000 01），and OPG mRNA expression （SMD=11.81，95% CI=7.49-16.13，P<0.000 01），promoting OPG protein production （SMD=4.95，95% CI=3.09-6.81，P<0.000 01），reducing serum RANKL（SMD=-4.88，95% CI=-6.01--3.75，P<0.000 01） and RANK levels （SMD=-7.30，95% CI=-9.53--5.07，P<0.000 01），and down-regulating RANKL （SMD=-6.22，95%CI=-8.95--3.49，P<0.000 01） and RANK mRNA （SMD=-3.18，95% CI=-6.19--0.18，P<0.05） expression and RANKL protein expression in bone tissue （SMD=-3.99，95% CI=-5.47--2.50，P<0.000 01）.ConclusionThe kidney-tonifying method has been proved to possess potential advantages in regulating the balance of OPG/RANK/RANKL signaling pathway in PMOP animal model. Nevertheless，more large-sample sized，properly designed，and high-quality animal experiments are still needed for further verification.
Abstract：ObjectiveTo systematically evaluate the efficacy of oral Chinese herbal prescriptions combined with transcatheter arterial chemoembolization （TACE） against primary hepatic carcinoma （PHC） and screen the basic Chinese herbs，in order to provide certain reference for clinical medication.MethodThe randomized controlled trials concerning the treatment of PHC with oral Chinese herbal prescriptions plus TACE were retrieved from CBM，China National Knowledge Infrastructure （CNKI），Chongqing Weipu Database for Chinese Technical Periodicals （VIP），and Wanfang Data Knowledge Service Platform.The quality of the included trials was evaluated by Cochrane handbook，and the Meta-analysis was performed using RevMan 5.3.The enumeration data were expressed by odds ratio （OR），the measurement data by mean difference （MD） or standardized mean difference （SMD），and the effect size by 95% confidence interval （CI）.The data of oral Chinese herbal prescriptions involved in trials were sorted out and subjected to association rule analysis and frequency analysis based on the Traditional Chinese Medicine Inheritance Support System （TCMISS），for exploring the basic Chinese herbs and their dosages against PHC.ResultA total of 75 randomized controlled trials were included，involving 7 406 cases. As revealed by the Meta-analysis，oral Chinese herbal prescriptions combined with TACE was significantly better than TACE alone in improving the short-term curative effect ［OR=2.05，95%CI（1.83，2.29）］，decreasing alpha fetoprotein （AFP） ［MD=-59.02，95%CI（-79.03，-39.01）］，ameliorating liver function ［SMD=-1.23，95%CI（-1.58，-0.88）］，boosting immunity ［SMD=1.08，95%CI（0.84，1.32）］，adjusting Karnofsky Performance Status （KPS） scale score ［OR=2.7，95%CI（1.11，11.02）］，elevating survival rate ［OR=2.31，95%CI（1.96，2.71）］，and reducing adverse reactions ［OR=0.38，95%CI（0.34，0.43）］.Data mining results showed that the basic Chinese herbs against PHC were Bupleuri Radix，Paeoniae Alba Radix，Atractylodis Macrocephalae Rhizoma，Poria，and Glycyrrhizae Radix et Rhizoma，with their clinical dosages listed as follows：6-15 g for Bupleuri Radix，10-15 g for Paeoniae Alba Radix，9-15 g for Atractylodis Macrocephalae Rhizoma，10-15 g for Poria，and 3-10 g for Glycyrrhizae Radix et Rhizoma.ConclusionThe oral Chinese herbal prescriptions combined with TACE produce better effects in treatment of PHC as compared with TACE alone.These five basic Chinese herbs have anti-cancer effect，and their dosages are within the ranges stipulated in 2020 edition of Chinese Pharmacopoeia.This Meta-analysis has provided certain reference for clinical medication.
Abstract：ObjectiveAuxiliary Verse on Drugs and methods for Zang-Fu Organs （Fuxingjue Wuzangyongyao Fayao） written by TAO Hong-jing from the Liang dynasty covered many contents from a missing prescription book Classic of Decoction （Tangye Jingfa）， including a map revealing the compatibility principle of Chinese herbs， namely the Tangye Jingfatu. Represented by a centrosymmetric pentagon， the map describes a unique theoretical system for deficiency-excess syndrome differentiation of five Zang organs （liver， heart， spleen， lung and kidney） and the tonification-purgation and compatibility theory of five flavors （pungent， salt， sweet， sour and bitter）. Each Zang organ fixedly corresponds to one "property" flavor （purgation）， one "function" flavor （tonification） and one "transformation" flavor （harmonization） resulting from the combination of the former two. For example， the liver can be purged by sour， tonified by pungent， and moderated by sweet transformed by the combination of sour with pungent. The heart can be purged by bitter， tonified by salt， and astringed by sour transformed by the combination of bitter with salt. The spleen can be purged by pungent， tonified by sweet， and dried by bitter transformed by the interaction between pungent and sweet. The lung can be purged by salt， tonified by sour， and dispersed by pungent transformed by the combination of salt with sour. The kidney can be purged by sweet， tonified by bitter， and moistened by salt transformed by the combination of sweet with bitter. This study selected appropriate mathematical tools to analyze the fixed relationship between "property" flavor， "function" flavor and "transformation" flavor among the five Zang organs in “Tangye Jingfatu” and establish a mathematical model revealing the compatibility-transformation relationship among five flavors.MethodBased on the group representation of five elements （wood， fire， earth， metal， and water）， the correlations of "property" flavor， "function" flavor， and "transformation" flavor with five elements’ generation-restriction were deduced based on matrix calculation and group theory. The three-dimensional calculation method for vector product was expanded for establishing the mathematical operator of five flavors' compatibility-transformation.ResultandConclusionThere is a mapping relationship of the purging， tonifying， and harmonizing functions represented by the "property" flavor， "function" flavor， and "transformation" flavor of the five zang organs in the "Tangye Jingfatu" with the five elements' generation-restriction. The established mathematical operator contributes to explaining the fixed collocations of five flavor transformation. Based on such algorithm， the tonifying and purging characteristics of five flavors in 10 representative classic prescriptions have been clearly expounded.
Abstract：Constipation is a common functional gastrointestinal disease with obstructive constipation， prolonged defecation time， or difficulty in defecation as the main symptoms， its prevalence is increasing year by year. Copying the animal model that is highly consistent with the clinical characteristics of traditional Chinese medicine （TCM） and western medicine is the basis for the related research on constipation. Based on the clinical characteristics， etiology and pathogenesis， clinical diagnostic criteria and TCM syndrome differentiation of constipation， a comparative analysis of the anastomosis between common animal models and clinical symptoms was carried out. Existing animal models of constipation were mostly drug induction and non-pharmaceutical intervention methods， and the modeling method was relatively simple， which was different from the multi-factors of constipation in clinical practice， and the evaluation indicators were mainly charcoal-powder propelling rate， defecation function， colon tissue and serum biochemical indicators. TCM syndrome differentiation lacked perfect evaluation system， such as laborious defecation， palpitation， shortness of breath， forgetfulness， cold pain in the abdomen， and other indicators that had not been embodied in the animal models， and the observation indicators were inconsistent with the diagnostic criteria of TCM. The same pathogenic factor could cause different syndromes， such as shortness of breath and fatigue， hot flashes and night sweats in the diagnostic criteria of TCM. Therefore， it is necessary to further explore the animal model of constipation that is highly consistent with the clinical characteristics of TCM and western medicine， in order to promote clinical treatment of constipation.
Keywords：constipation;modeling method;integrated traditional Chinese and western medicine;diagnostic criteria;clinical syndromes;biochemical index;gastrointestinal diseases
Abstract：In recent years， with the change in life style， social environment， and national childbearing policy， the proportion of high-risk pregnant women has increased significantly， triggering the spectrum of obstetric diseases to constantly change， which has brought new challenges to the diagnosis and treatment of obstetrics. Traditional Chinese medicine （TCM） has been proved effective in dealing with a variety of obstetric diseases， and various treatment methods are available， which can serve as alternative means for solving refractory obstetric diseases. However， most obstetric clinicians are currently less aware of the therapeutic effects of TCM， which has significantly hindered its participation in clinical treatment. Therefore， the China Association of Chinese Medicine （CACM） organized the outstanding young obstetricians of TCM and western medicine to discuss 15 obstetric diseases responding specifically to TCM or integrated TCM and western medicine， including hyperemesis gravidarum， threatened abortion， ectopic gestation， cough during pregnancy， pregnancy-induced hypertension syndrome， maternal-fetal ABO incompatibility， postpartum hypogalactia， residual pregnancy tissue in uterine cavity， puerperal infection， pantalgia after childbirth， hematoma/undesirable healing after caesarean section， postpartum urinary retention， ileus after cesarean section， pelvic floor dysfunction， and postnatal depression. The suggestions for their treatment with TCM or integrated TCM and western medicine were also proposed， aiming to provide patients with effective and personalized treatments in clinical practice and improve the diagnosis and treatment effects of obstetric diseases， thus benefiting the public. At the same time， more obstetrical clinicians are expected to understand the therapeutic effects and advantages of TCM and draw on the strengths of both TCM and western， thereby promoting the establishment of an obstetric diagnosis and treatment system with Chinese characteristics.
Keywords：obstetrics;diseases responding specifically to traditional Chinese medicine （TCM）;expert consensus
Abstract：Cancer is a threat to human health. New treatments for cancer can significantly prolong the survival time of patients， but fail to improve the adverse reactions induced by chemoradiotherapy. Improving patient outcomes still requires the effort of cancer researchers. In recent years， the anti-tumor effects of active components from Chinese herbs have received wide attention. Kaempferol， a flavonoid mainly found in the medicinal plant Kaempferia galanga， can be used to treat obesity， cardiovascular diseases， diabetes and other diseases. It has also exhibited good efficacy in inhibiting the occurrence and development of liver cancer， colon cancer， lung cancer， ovarian cancer， and other malignant tumors. Kaempferol mainly exerts the anti-cancer effect by inducing apoptosis. Specifically， it promotes the production of intracellular reactive oxygen species （ROS） and triggers cell apoptosis through the mitochondrial pathway. Besides， it is capable of interfering with the cancer cell cycle， causing most cancer cells to arrest in the G2/M phase， and inducing cell autophagy， a programmed cell death， thus inhibiting cell migration and invasion and angiogenesis， synergistically improving chemotherapeutic drug efficacy， and reducing adverse effects. Kaempferol acts on a series of intracellular and extracellular targets to participate in the regulation of tumor cell signaling pathways， involving phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， epidermal growth factor receptor （EGFR）， mitogen activated protein kinase （MAPK）， and Wnt signaling pathways， with the PI3K/Akt signaling pathway being most significant. In addition， kaempferol also plays an important regulatory role in tumor epigenetics. This paper reviewed the anti-tumor effect and mechanism of kaempferol， aiming to provide reference for in-depth study on its prevention and treatment of tumor and the development of new anti-tumor drugs.
Keywords：kaempferol;anti-tumor;mechanism of action;apoptosis
Abstract：Sjögren syndrome （SS） is a chronic autoimmune disease characterized by immune cell infiltration and progressive destruction of salivary and lacrimal glands. It not only affects the lacrimal and salivary glands， manifested as dry eyes and dry mouth， but also involves heart， lung，kidney，and central nervous system， seriously affecting human physical and mental health. Although western medicine has made extensive and in-depth research on the diagnosis and treatment of this disease in recent years，there is no effective treatment targeting the potential causes. Chinese medicine emphasizes the concept of holism，treatment and prescription formulation based on syndrome differentiation， and effect exertion via multiple targets，multiple levels，and multiple pathways，exhibiting great advantages in the treatment of SS. This paper reviews the mechanisms of Chinese medicine in treating SS from the perspectives of immunity regulation，aquaporin up-regulation， and anti-oxidative stress reported in the related literature，so as to provide more theoretical basis for the research and clinical treatment of SS.
Keywords：Sjögren syndrome （SS）;traditional Chinese medicine;immunity regulation;aquaporins;anti-oxidation
Abstract：Lung cancer is a malignant tumor with high incidence and high mortality， posing a great threat to human health. Neovascularization may be one of the important mechanisms of lung cancer. The growing lung cancer cells can obtain necessary nutrients from the newly formed blood vessels， thereby causing the spread and metastasis of lung cancer. Nowadays， anti-angiogenic drugs are commonly used in western medicine in addition to surgery，radiotherapy， chemotherapy， and immunotherapy. However， the resulting adverse reactions such as thrombosis， hypertension， diarrhea， and cardiotoxicity have seriously affected the quality of life of patients. As the recognition of angiogenesis deepens， the selection of lung cancer treatment options has become a research hotspot and difficulty in the field of lung cancer treatment. In traditional Chinese medicine（TCM）， angiogenesis is believed to fall into the category of “collateral disease”. The invasion of external pathogens and deficiency of healthy Qi will cause visceral dysfunction， which can be gradually followed by Qi obstruction and blood stasis and phlegm-turbidity congesting the collaterals. As a result， the collateral function will be damaged， providing favorable conditions for the occurrence of lung cancer. More and more modern studies have confirmed that TCM is able to inhibit angiogenesis in the lung cancer， thereby resisting the tumor. In addition， by virtue of the unique advantages， TCM effectively reduces adverse reactions， enhances the efficacy， and improves the living conditions of patients. Moreover， it can synergize with other western medicine therapies in the treatment of lung cancer， exhibiting a wide application prospect. This paper summarizes the mechanisms of TCM in inhibiting angiogenesis of lung cancer reported in relevant experimental research， hoping to provide reference for the optimization of clinical treatment strategies for lung cancer.
Keywords：lung cancer;angiogenesis;traditional Chinese medicine;mechanism of action