Abstract：ObjectiveTo observe the regulatory effect of Xiao Qinglongtang and its ingredients on lung water transport-related proteins， and to explain the biological connotation of lung governing water movement， based on which the regulatory mechanism of Xiao Qinglongtang will be explored.MethodAccording to the composition rules of classical formula， Xiao Qinglongtang （11.22 g·kg-1）， Guizhi Gancao （2.70 g·kg-1）， Shaoyao Gancao （2.70 g·kg-1）， Jiangxinwei （3.90 g·kg-1）and Banxia Muahuang （0.032 7 g·kg-1） were prepared. The pathological model of syndrome of cold fluid accumulated in lung of rats was established by the "coldness of body + drinking cold + cold bath" method， and Xiao Qinglongtang and its ingredients were administrated to intervene with the model rats. Lung function parameters of forced vital capacity （FVC）， functional residual capacity （FRC）， mean mid-expiratory flow （MMEF）， inspiratory time （tI）， and inspiratory time （tE） were determined by lung function analyzer. Hematoxylin and eosin （HE） staining was used to observe the changes in pathological morphology. The expression of aquaporin （AQP）1， AQP5， epithelial sodium channel α subunit（α-ENaC） and Na+-K+-ATPase in lung tissues of rats， the content of tumor necrosis factor -α（TNF-α）， the mRNA expression of cyclic adenosine monophosphate （cAMP）， protein kinase A （PKA） and cAMP-response element binding protein （CREB）， and the protein expression of cAMP， PKA， CREB， and phosphorylated-CREB （p-CREB） were detected by immunohistochemistry （IHC）， enzyme-linked immunosorbent assay （ELISA）， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and Western blot， respectively.ResultCompared with normal group， functions of FVC， FRC and MMEF in model group were significantly decreased (P<0.01)， and the time of tI and tE was significantly prolonged （P<0.05，P<0.01）. The content of TNF-α in lung tissue was significantly increased （P<0.01）. The mRNA and protein expressions of cAMP， PKA and CREB in lung tissue were significantly decreased （P<0.01）. The expression of AQP5 and α-ENAC in lung tissue decreased significantly. The alveolar cavity of rats was filled with edema fluid， surrounding tissue hyperemia， inflammatory cell infiltration, bronchial mucosa epithelial adhesion. Compared with model group， Xiao Qinglongtang and its fangyuan group could significantly enhance the FVC, FRC and MMEF functions of model rats （P<0.05，P<0.01）， and tI and tE time were shortened （P<0.05，P<0.01）. The content of TNF-α in lung tissues of Xiao Qinglongtang group， Guizhi Gancao group and Banxia Mahuang group was significantly decreased （P<0.01）. The mRNA expressions of cAMP， PKA and CREB in Xiao Qinglongtang group were significantly up-regulated （P<0.01）， and the mRNA expressions of cAMP and PKA in Guizhi Gancao， Jiangxinwei and Banxia Mahuang groups were significantly up-regulated （P<0.01）. The protein expressions of cAMP， PKA and CREB in Xiao Qinglongtang group， Guizhi Gancao group， Jiangxinwei group and Banxia Mahuang group were significantly up-regulated （P<0.01）， and the protein expression of CREB in Shaoyao Gancao group was significantly up-regulated（P<0.05）. Xiao Qinglongtang could up-regulate the positive expression of AQP5 and α-ENAC， and Guizhi Gancao group could up-regulate the positive expression of α-ENAC. Xiao Qinglongtang and its fangyuan can reduce the lung edema， inflammatory cell infiltration and bronchial mucosal adhesion of model rats.ConclusionXiao Qinglongtang and its ingredients can reduce lung edema and inhibit inflammation by improving the expression of lung water transport-related proteins AQP1， AQP5， and α-ENaC through cAMP/PKA pathway， thereby restoring the lung functions in rats with syndrome of cold fluid accumulated in lung. Na+-K+-ATPase may play an auxiliary role in the regulation of lung water transport. This provides a certain objective basis for preliminarily elucidating the connotation of lung governing water movement from the perspective of lung water transport-related proteins.
Keywords：Xiao Qinglongtang;lung governing water movement;aquaporins;epithelial sodium channel;Na+-K+-ATPase
Abstract：ObjectiveTo observe the effects of Suanzaoren Tang on the behavior， growth-associated protein-43 （GAP-43）， postsynaptic density protein-95 （PSD-95）， and synaptophysin Ⅰ （SynⅠ） of insomniac rats induced by p-chlorophenylalanine （PCPA）， and to investigate the mechanism of Suanzaoren Tang in improving the behavior of the insomniac rats.MethodSeventy-two SD rats were randomly assigned into 6 groups （12 rats in each group）： control group （normal saline）， PCPA （0.35 g·kg-1·d-1） group， estazolam （2.7×10-4 g·kg-1·d-1） group， and low-， medium-， and high-dose （3.25， 7.5， 15 g·kg-1·d-1， respectively） Suanzaoren Tang groups. The rat model of insomnia was established by intraperitoneal injection of PCPA and then the rats were administrated with corresponding drugs for 7 continuous days. The Morris water maze and Y-maze were used to test the learning and memory functions， and the open field to test anxiety. Histopathological changes in the hippocampus were observed via hematoxylin-eosin （HE） staining. The mRNA and protein levels of GAP-43， PSD-95， and SynⅠ in hippocampus were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR)， Western blot， and immunohistochemistry （IHC）.ResultCompared with the control group， the PCPA group showcased long escape latency， shortened time in the quadrants， and decreased times of crossing the platform in Morris water maze， decreased alternation correct rate was significantly Y-maze， and increased distance， mean velocity， and time in center of the open field test （P<0.01）. Furthermore， the PCPA-treated rats showed obvious pathological damage in the hippocampus and down-regulated mRNA and protein levels of GAP-43， PSD-95， and SynⅠ in hippocampus （P<0.01）. Compared with the PCPA group， the treatments with estazolam and different doses of Suanzaoren Tang improved the rat performance in Morris water maze， Y-maze， and open field test （P<0.05， P<0.01）， alleviated the hippocampal damage， and up-regulated the mRNA and protein levels of GAP-43， PSD-95， and SynⅠ （P<0.05， P<0.01）.ConclusionSuanzaoren Tang may alleviate the learning and memory disorders and anxiety in PCPA-induced insomnia rat model by up-regulating the mRNA and protein levels of hippocampal synaptic plasticity-associated proteins GAP-43， PSD-95， and SynⅠ.
Abstract：ObjectiveTo observe the protective effect of Chaihu Jia Longgu Mulitang （CLMT） on dopaminergic neurons in Parkinson's disease with depression （PDD） model rats， and to explore the mechanism based on adenosine monophosphate-activated protein kinase/mammalian target of rapamycin （AMPK/mTOR） signaling pathway.MethodAmong the 80 male SD rats， 10 were randomly selected as normal group and the rest were treated with long-term low-dose subcutaneous injection of rotenone in the neck and back combined with chronic unpredictable mild stress （CUMS） to establish PDD rat model. The successfully modeled PDD rats were randomly divided into model group， western medicine group （madopar 0.032 g·kg-1+fluoxetine hydrochloride 0.002 g·kg-1）， CLMT low-dose， medium-dose and high-dose groups （5， 10 and 20 g·kg-1）， 10 rats in each group. Normal group and model group were administrated with the same amount of normal saline by gavage for 4 consecutive weeks. Behavioral changes of rats in each group were evaluated by open field test and pole climbing test. The content of dopamine （DA） and 5-hydroxytryptamine （5-HT） in cerebrospinal fluid was determined by high performance liquid chromatography （HPCL）. The pathological changes of dopaminergic neurons in substantia nigra of rats were observed by hematoxylin-eosin （HE） staining. The positive expression of tyrosine hydroxylase （TH） and expression of α-synuclein in substantia nigra were detected by immunohistochemistry （IHC） and immunofluorescence （IF）， repsectively. The protein expression of microtubule-associated protein 1 light chain 3 （LC3）， adenosine monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mammalian target of rapamycin （mTOR） and phosphorylated mTOR （p-mTOR） was detected by Western blot.ResultCompared with the conditions in normal group， the total horizontal distance and the activity time in the central region in open field test and the content of DA and 5-HT in cerebrospinal fluid were decreased （P<0.05， P<0.01）， and the time of pole climbing was shortened （P<0.01）， with increased score （P<0.01） in model group. Compared with model group， CLMT high-dose group and western medicine group increased the total horizontal distance and activity time in the central region and the content of DA and 5-HT （P<0.05， P<0.01）， and extended the time of climbing pole （P<0.05）， with decreased score （P<0.05， P<0.01）. Compared with those in normal group， the number of dopaminergic neurons in the substantia nigra was reduced， with narrowed and loosely arranged cell body. The fluorescence expression of α-synuclein was enhanced （P<0.01）， and the positive expression of TH was decreased （P<0.01） in model group. Compared with model group， CLMT high-dose group and western medicine group showed elevated number of dopaminergic neurons in the substantia nigra， with enlarged cell body， and decreased fluorescence expression of α-synuclein， and enhanced the positive expression of TH （P<0.05， P<0.01）. Compared with normal group， model group had lowered expression of LC3Ⅱ/Ⅰ， p-AMPK/AMPK in striatum （P<0.05， P<0.01） and increased expression of p-mTOR/mTOR （P<0.01）. Compared with those in model group， LC3Ⅱ/Ⅰ and p-AMPK/AMPK expression were increased （P<0.05， P<0.01） and p-mTOR /mTOR expression was decreased （P<0.01） in CLMT high-dose group and western medicine group.ConclusionCLMT exerts a neuroprotective effect by inhibiting rotenone neurotoxicity. It enhances the level of DA， and thus improves the depression condition in rats with Parkinson's disease. The underlying mechanism may be related to the regulation of AMPK/mTOR signaling pathway， activation of autophagy， and promotion of degrading α-synuclein.
Keywords：Chaihu Jia Longgu Mulitang （CLMT）;Parkinson's disease with depression （PDD）;adenosine monophosphate-activated protein kinase/mammalian target of rapamycin （AMPK/mTOR） signaling pathway;autophagy;neuroprotective effect
Abstract：ObjectiveBy observing the effect of Xiaoluowan on phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin complex 1 （mTORC1） pathway in experimental goiter rats， this study aims to explore its therapeutic effect on experimental goiter rats.MethodSixty 5-month-old SD rats of SPF grade were purchased， half males and half females， of which 10 were used as a normal group， and the remaining rats were administrated with propylthiouracil （PTU） solution to induce nodular goiter. After successful modeling， rats were randomly divided into a model group， levothyroxine sodium tablets group， Xiaoluowan low-dose group， medium-dose group， and high-dose group， ten rats each. The levothyroxine sodium tablets group was given 15 μg·kg-1 levothyroxine sodium tablets by gavage. The Xiaoluowan low-， medium-， and high-dose groups were given （ig） Xiaoluowan low-dose （10 g·kg-1）， medium-dose （20 g·kg-1）， and high-dose （30 g·kg-1） Xiaoluowan， and the normal group and model group were administered （ig） with the same volume of 0.9% sodium chloride solution. Four weeks after the intervention， rats were sacrificed by routine intraperitoneal anesthesia using 5% phenobarbital. Subsequently， the histopathology was observed under a microscope， and serum thyroid hormone levels were measured using a Roche electrochemiluminescence immunoassay analyzer. Serum cytokines were detected by enzyme-linked immunosorbent assay （ELISA）， and neurotransmitters were measured using a high-performance liquid chromatograph. The protein level of PI3K/Akt/mTORC1 pathway was determined by Western blot.ResultAs compared with the normal group， the levels of basic fibroblast growth factor （bFGF）， vascular endothelial growth factor （VEGF）， insulin-like growth factor （IGF-1）， 5-hydroxytryptamine （5-HT）， and thyroid stimulating hormone （TSH） were increased， and PI3K， Akt， and mTORC1 protein levels were up-regulated in the model group， while the levels of norepinephrine （NE）， triiodothyronine （T3）， tetraiodothyronine （T4）， free triiodothyronine （FT3）， and free thyroid hormone （FT4） were decreased （P<0.05）. Compared with the model group， the levothyroxine sodium tablets group， and Xiaoluowan low-， medium-， and high-dose groups exhibited reduced levels of bFGF， VEGF， IGF-1， 5-HT， and TSH， and down-regulated PI3K， Akt， and mTORC1 protein levels， and increased NE， T3， T4， FT3， and FT4 levels （P<0.05）.ConclusionXiaoluowan may act on the PI3K/Akt/mTORC1 signaling pathway to play its role in the treatment of nodular goiter， and it is dose-dependent.
Abstract：ObjectiveTo study the effect of Jinlida granules on visceral fat accumulation and its induced inflammatory response in prediabetic rats.MethodMale SD rats were randomly divided into normal group， model group， Jinlida low-dose group （1.5 g·kg-1）， Jinlida high-dose group （3.0 g·kg-1） and atorvastatin group （10 mg·kg-1）. Prediabetic rat model was established using high-carbohydrate， high-fat diet combined with low-dose streptozotocin （STZ） by multiple small-dose intraperitoneal injections. After 8 weeks of modeling and drug intervention for 13 consecutive weeks， body weight， oral glucose tolerance test（OGTT）， fasting blood glucose （FBG）， fasting insulin （FINS）， insulin resistance index （HOMA-IR）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C） were measured in each group of rats. The content of visceral fat was quantified by micro-computed tomography （Micro-CT）. Hematoxylin-eosin staining （HE） was used to observe the pathological changes of fat cells. The levels of tumor necrosis factor-α （TNF-α） and interleukin- 6 （IL-6） in rat visceral fat and serum were determined by enzyme linked immunosorbent assay （ELISA）. The expression of macrophage marker CD68 in visceral fat was detected by immunofluorescence and Western blot.ResultCompared with normal group， model group had increased oral glucose tolerance， FBG， FINS， HOMA-IR， TC， LDL-C （P<0.01）， elevated body weight and visceral fat accumulation （P<0.05， P<0.01）， enhanced CD68 protein expression and TNF-α and IL-6 levels （P<0.01）， decreased HDL-C （P<0.01）， and abnormal hypertrophy of adipocytes. Compared with model group， Jinlida high- and low-dose groups lowered oral glucose tolerance， HOMA-IR， TC and LDL-C （P<0.05， P<0.01）， body weight and visceral fat accumulation （P<0.05）， and CD68 protein expression and TNF-α and IL-6 levels （P<0.05， P<0.01） and lessened hypertrophy of fat cells.ConclusionJinlida can improve the insulin resistance in prediabetic rats by reducing visceral fat accumulation and its induced inflammatory response， which provides a new pharmacological basis for clinical treatment of prediabetes by Jinlida granules.
Abstract：ObjectiveTo explore the regulatory effect of Gouqi chewable tablets on innate and adaptive immunity in normal mice and its antioxidant activity in vitro and in vivo.MethodThe effects of low-， medium-， and high-dose groups （0.25， 0.5， 1.5 g·kg-1） on the immune function of normal mice were observed by carbon clearance test， immune organ index test， serum hemolysin test， ConA-induced splenic lymphocyte proliferation test， and natural killer cell （NK cell） activity test. The effects of Gouqi chewable tablets on the antioxidant capacity in vivo were determined by detecting the content of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） in mice serum. The in vitro antioxidant activity of Gouqi chewable tablets was detected by 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） diammonium salt （ABTS）， 2，2-diphenyl-1-picrylhydrazyl （DPPH）， and hydroxyl radical scavenging tests.ResultCompared with the blank control group， the low-， medium-， and high-dose groups of Gouqi chewable tablets improved the viability of NK cells， the proliferation of splenic lymphocytes， and the level of serum hemolysin antibody in mice （P<0.05）. The high-dose group increased the thymus index， spleen index， and phagocytic function of macrophages （P<0.05， P<0.01）. As compared with the blank control group， the activity of GSH-Px in mice serum in the medium-dose group was increased （P<0.05）， and the content of MDA in mice serum in the high-dose group was decreased （P<0.05）. In in vitro antioxidant tests， the median inhibitory concentration （IC50） values of Gouqi chewable tablets were 1.64±0.20， 2.04±0.03， and 10.27±0.03 g·L-1 by the DPPH， ABTS， and OH- free radical method， respectively. Those results indicated that Gouqi chewable tablets have good antioxidant effects in vitro.ConclusionGouqi chewable tablets can enhance the immune function of mice with good antioxidant effects.
Abstract：ObjectiveTo study the virulence and biofilm inhibition effect of Fufang Huangbai Fluid Paint （FFHBFP） on methicillin-resistant Staphylococcus aureus （MRSA）， and to explore the antibacterial effect of FFHBFP on MRSA， which provides a theoretical basis and reference for clinical medication.MethodFirstly， the microdilution method and time–growth curve were used to determine the minimum inhibitory concentration （MIC） of FFHBFP and vancomycin （VAN） against MRSA and the effect on bacterial growth. The effects of FFHBFP and VAN on the inhibition of MRSA virulence factor lipase and restoration of hydrogen peroxide （H2O2） sensitivity were detected under sub-minimum inhibitory concentration （sub-MIC）. The inhibitory effect of FFHBFP and VAN on MRSA biofilm formation and maturation was detected by the microplate method. The morphological changes of mature biofilms before and after administration were observed under a scanning electron microscope （SEM）. Real-time polymerase chain reaction （Real-time PCR） was utilized to detect the effect of 50.600 g·L-1 concentration of FFHBFP on the expression of MRSA virulence gene crtM and biofilm-forming genes fnbA and icaA. Finally， molecular docking technology was used to predict the mechanism of potential antibacterial active ingredients of FFHBFP in inhibiting the virulence and biofilm of MRSA.ResultThe MIC of VAN was 2 mg·L-1， and VAN below 1 mg·L-1 exerted no effect on MRSA growth. The MIC of FFHBFP was not determined， while the 101.200-202.400 g·L-1 original solution inhibited MRSA growth. Compared with the blank group and the VAN group， sub-MIC （25.300-50.600 g·L-1 original solution） inhibited lipase and recovered MRSA sensitivity to H2O2 （P<0.01）. The results of the microplate method showed that FFHBFP （25.300-202.400 g·L-1 original solution） inhibited biofilm formation and maturation （P<0.05， P<0.01）. The SEM exhibited that FFHBFP made the structure of biofilm loose and the size of the bacteria varied. FFHBFP at 50.600 g·L-1 concentration can inhibit the expression of related virulence genes and biofilm-forming genes （P<0.05， P<0.01）， and molecular docking results also showed that the main antibacterial active ingredients in FFHBFP have good binding ability to the target.ConclusionFFHBFP that cannot directly kill MRSA exerts clinical efficacy by impairing virulence expression， biofilm formation， and other pathogenic properties.
Abstract：ObjectiveTo analyze the transcriptome characteristics of Xianlian Jiedu prescription （XLJDP） in the intervention of colorectal carcinoma by high-throughput cDNA-sequencing （RNA-seq）.MethodNinety male C57BL/6 mice were randomly divided into the control group， colorectal carcinoma due to dampness， heat， stasis， and toxin model group， and XLJDP group， with 30 mice in each group. Mice in the model group and XLJDP group were fed a high-fat diet and provided with azoxymethane and dextran sodium sulfate （AOM/DSS） for inducing colorectal carcinoma. Those in the XLJDP group were further treated with intragastric administration of 12.9 g·kg-1 XLJDP since the day of modeling for 112 days. The colorectal tissues were collected from each group 4 h after the last drug treatment and stained with hematoxylin-eosin （HE） and methylene blue for observing the pathological changes. The total RNA was extracted from colorectal tissues for RNA-Seq-based transcriptome profiling， followed by gene oncology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment analysis and the screening and verification of differentially expressed genes.ResultCompared with the model group， XLJDP significantly relieved the colorectal congestion and edema and decreased tumor number and volume in mouse colorectal tissues. The methylene blue staining results indicated that XLJDP significantly suppressed the development of aberrant crypt foci （ACF，P<0.01）. As revealed by HE staining， XLJDP significantly alleviated the injury and dysplasia of colorectal tissues. Transcriptome analysis identified 615 differentially expressed genes （446 up-regulated and 169 down-regulated） between the model group and the blank group and 54 differentially expressed genes （29 up-regulated and 25 down-regulated） between the XLJDP group and model group. XLJDP mainly affected the expression of NIMA-related protein kinase 7 gene （Nek7， P<0.01）， Mucin 16 （Muc16， P<0.01）， SiahE3 ubiquitin protein ligase family member 3 （Siah3， P<0.01）， regenerating islet-derived protein 3-gamma （Reg3g， P<0.01）， RNA polymerase Ⅱ elongation factor-associated factor 2 （Eaf2， P<0.01）， transforming growth factor‐alfa gene （TGF-α， P<0.05）， secretoglobin family 1A member 1 （Scgb1a1， P<0.05）， family with sequence similarity 227 member B （Fam227B， P<0.05）， cytochrome P450 family 2 subfamily c polypeptide 40 （Cyp2c40， P<0.01）， and ankyrin repeat and EF-hand domain containing protein 1 （Ankef1， P<0.05）. Enrichment analysis showed that intestinal epithelial cell proliferation， metabolism of xenobiotics by cytochrome P450， and arachidonic acid metabolism signaling pathway were significantly enriched.ConclusionXLJDP is able to interfere with colorectal tumorigenesis and development due to dampness， heat， stasis， and toxin in mice， which has been proved by transcriptome analysis to be related to the regulation of metabolism-related pathways.
Abstract：ObjectiveTo explore the effect of Xianlian Jiedu prescription （XLJDP） on the proliferation and glycolysis of human colorectal cancer HCT-116 cells and the underlying mechanism.MethodHCT-116 cells were cultured with XLJDP and then the survival rate was examined by methyl thiazolyl tetrazolium （MTT） assay. The effect on the HCT116 cell proliferation was detected by colony formation assay and 5-ethynyl-2′-deoxyuridine （EDU） incorporation assay. The amount of glucose consumed by HCT-116 cells was measured by glucose test kit， and the amount of produced lactic acid was determined by lactic acid test kit 48 h after the treatment with XLJDP. The expression of glycolysis-related proteins mammalian target of rapamycin （mTOR）， phosphorylated mTOR （p-mTOR）， glucose transporter 1 （GLUT1）， and lactate dehydrogenase （LDHA） was detected by Western blot.ResultThe half-maximal inhibitory concentration （IC50） of XLJDP against HCT-116 cells was 6.82 g·L-1. Compared with the blank group， XLJDP （1.625， 3.25， 6.50 g·L-1） inhibited the proliferation of HCT-116 cells （P<0.05， P<0.01）. Moreover， compared with the blank group， XLJDP （1.625， 3.25， 6.50 g·L-1） suppressed glucose uptake and lactic acid production in a dose-dependent manner （P<0.05， P<0.01）. The expression of p-mTOR/mTOR， LDHA， and GLUT1 was down-regulated by XLJDP （P<0.05， P<0.01）.ConclusionXLJDP can significantly inhibit the proliferation and the Warburg effect of glycolysis in colorectal cancer cells by regulating the mTOR signaling pathway and the down-regulating the expression of LDHA， GLUT1， and other key proteins and enzymes in glycolysis.
Abstract：ObjectiveTo study the effect of Xianlian Jiedu prescription （XLJDP） on the activation of nuclear transcription factor-κB （NF-κB） signaling pathway induced by bromodomain-containing protein 4 （Brd4） in hypoxic microenvironment and to explore its mechanism in inhibiting the proliferation of colorectal cancer HT-29 cells.MethodThe human colorectal cancer HT-29 cells were cultured in a hypoxic incubator or normoxia incubator and treated with XLJDP at 0.8，1，1.2，1.6，3.2，6.4，and 12.8 g·L-1 for 48 h， respectively. Following the detection of cell vitality using methyl thiazolyl tetrazolium （MTT） colorimetry， the effects of XLJDP （1.25，2.5，and 5 g·L-1） on the cell mitochondrial membrane potential were determined using a fluorescent probe （JC-1）， and the apoptosis of colorectal cancer HT-29 cells was detected by flow cytometry. The cell colony formation assay and 5-ethynyl-2'-deoxyuridine （EDU） staining were conducted to test the proliferation of colorectal cancer HT-29 cells. The Western blot was carried out to measure the expression levels of Brd4 and its downstream relevant proteins such as c-Myc and hexamethylene bisacetamide-inducible protein 1 （HEXIM1）， as well as the effects of XLJDP on related proteins in the NF-κB signaling pathway.ResultCompared with the blank control group， XLJDP at 0.8，1，1.2，1.6，3.2，6.4，and 12.8 g·L-1 inhibited the vitality of colorectal cancer HT-29 cells （P<0.05 ， P<0.01）， with the median inhibitory concentration （IC50） under the hypoxic condition higher than that under the normoxia condition. Compared with the blank control group， XLJDP at 1.25，2.5，and 5 g·L-1 significantly decreased the mitochondria membrane potential， enhanced the apoptosis （P<0.05，P<0.01）， and lowered the number of cell colonies and also the EDU-positive cells （P<0.05， P<0.01）. The results of Western blot showed that compared with the blank control group， XLJDP at 1.25，2.5，and 5 g·L-1 down-regulated Brd4， c-Myc， p-NF-κB p65， and p-IκBα protein expression to varying degrees and up-regulated the expression of HEXIM1 （P<0.05， P<0.01）.ConclusionIn the hypoxic microenvironment， XLJDP inhibits the proliferation of colorectal cancer HT-29 cells regulated by Brd4， which may be related to its inhibition of the activation of NF-κB signaling pathway.
Abstract：ObjectiveTo observe the effect of Xianlian Jiedu prescription （XLJDP） on the proliferation， apoptosis， and migration of cancer-relative endothelial （CRE） cells， and to decipher the mechanism of XLJDP in regulating angiopoietin2 （Ang2） to maintain CRE cell homeostasis and inhibit tumor neovascularization.MethodHuman umbilical vein endothelial cell line （HUVEC-c） was induced into CRE cells in the human colorectal cancer HCT-116 cell-conditioned medium. The CRE cells were assigned into the blank group， conditioned medium group， and XLJDP groups （1， 2， 3 g·L-1） and treated for 48 h. The proliferation of CRE cells was detected by methyl thiazolyl tetrazolium （MTT） colorimetry. The morphological changes of CRE cells were observed via an inverted microscope. The apoptosis rate was detected by flow cytometry. Wound healing test and Transwell migration assay were employed to detect the 2D/3D migration ability of CRE cells. The protein levels of vimentin， N-cadherin， matrix metalloproteinase-9 （MMP-9）， and Ang2 in CRE cells were measured by Western blot.ResultThe MTT results showed that the cell viability was higher in the conditioned medium group than in the blank group （P<0.05）. Compared with the conditioned medium group， XLJDP decreased the cell proliferation rate （P<0.01） and changed the cell morphology. The total apoptosis rates of all the XLJDP groups were higher than that of the conditioned medium group （P<0.01）. The 2D and 3D migration abilities of the conditioned medium group were higher than those of the blank group （P<0.05， P<0.01）. Compared with the conditioned medium group， XLJDP at all the concentrations weakened the 2D migration ability （P<0.01） and medium- and high-concentration XLJDP weakened the 3D migration ability （P<0.01）. The protein levels of N-cadherin， Vimentin， MMP-9， and Ang2 were up-regulated in the conditioned medium group compared with those in the blank group （P<0.05， P<0.01）. Compared with the conditioned medium group， XLJDP at all the concentrations down-regulated the protein level of Ang2 （P<0.05， P<0.01）， and medium- and high-concentration XLJDP down-regulated those of N-cadherin， vimentin， and MMP-9 protein （P<0.01）.ConclusionXLJDP may inhibit the proliferation， migration， differentiation， and apoptosis of CRE cells by down-regulating the expression of Ang2， inhibiting tumor neovascularization， and maintaining the cell homeostasis.
Keywords：Xianlian Jiedu prescription（XLJDP）;colorectal cancer;vascular homeostasis;angiopoietin2;conditioned medium
Abstract：ObjectiveTo investigate the effect of combined therapy of lung and intestine （Mahuangtang + Da Chengqitang） on lipopolysaccharide （LPS）-induced acute lung injury （ALI） in rats and its protective mechanism.MethodWistar rats were randomly divided into blank group， model group， low-， medium-， and high-dose groups with combined therapy of lung and intestine ， and dexamethasone group. LPS （10 mg·kg-1） was given （ip） to induce ALI in rats. The general state of rats in each group was observed and recorded. The body temperature of rats in each group was recorded 0-8 h after modeling by means of anal temperature measurement. Serum and lung tissues were collected 24 h after modeling. Serum levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， interleukin-10 （IL-10）， and arginase-1 （Arg-1） were determined by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein levels of nuclear factor kappa B p65 （NF-κB p65）， phosphorylated NF-κB p65 （p-NF-κB p65）， NF-κB inhibitor α （IκBα）， and phosphorylated IκBα （p-IκBα） in lung tissues of rats. The levels of classically activated （M1） macrophage marker CD80 and IL-1β and macrophage markers F4/80 and IL-10 were detected by double immunofluorescence.ResultCompared with the blank group， the model group showed increased body temperature and thermal response index （TRI）， elevated serum levels of pro-inflammatory factor TNF-α and IL-1β and anti-inflammatory factor IL-10 （P<0.01）， up-regulated protein levels of p-NF-κB p65 and p-IκBα in lung tissues （P<0.01）， and increased levels of F4/80， CD80， and IL-1β in lung tissues （P<0.01）. Compared with the model group， the lung-intestine combined treatment groups and the dexamethasone group exhibited decreased body temperature and TRI in rats （P<0.01）， declined serum levels of inflammatory factor TNF-α and IL-1β （P<0.05， P<0.01）， elevated serum levels of anti-inflammatory factor IL-10 and Arg-1 （P<0.05， P<0.01）， down-regulated protein levels of p-NF-κB p65 and p-IκBα in lung tissues （P<0.05， P<0.01）， decreased levels of CD80 and IL-1β， and increased levels of IL-10 in lung tissues （P<0.01）， while the level of F4/80 was not significantly changed.ConclusionThe combined therapy of lung and intestine can obviously alleviate the fever and inflammatory state of ALI rats， and the mechanism may be related to the inhibition of NF-κB inflammatory pathway and the polarization of lung tissue macrophages to anti-inflammatory phenotype.
Keywords：acute lung injury;combined therapy of lung and intestine;inflammation;nuclear factor kappa B （NF-κB）;macrophage polarization;Mahuangtang;Da Chengqitang
Abstract：ObjectiveTo explore the mechanism of the combined therapy of lung and intestine （Mahuangtang + Da Chengqitang） in alleviating pulmonary edema in rats with acute lung injury （ALI） induced by lipopolysaccharide （LPS）.MethodWistar rats were randomly divided into blank group， model group， low-， medium-， and high-dose groups with combined therapy of lung and intestine， and positive control group. LPS （10 mg·kg-1） was given （ip） to induce ALI in rats. After modeling， the blank group was given normal saline （25 mL·kg-1）， the combined therapy of lung and intestine treatment groups were given （ig） low- （5 g·kg-1）， medium- （7.5 g·kg-1）， and high-dose （10 g·kg-1） Mahuangtang and Da Chengqitang， and the positive control group was given dexamethasone （5 mg·kg-1）. Medications were administered 0， 8， and 16 h after LPS injection for 3 times. Then lung tissue and serum were collected after administration. The lung tissues were stained with haematoxylin-eosin （HE）， and the pulmonary edema score was evaluated. The dry/wet （D/W） weight ratio of lung tissues in each group was measured， and the content of serum vasoactive intestinal peptide （VIP） in rats was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein levels of aquaporin-1 （AQP1）， AQP5， VIP， cyclic adenosine monophosphate （cAMP）， phosphorylated protein kinase A （p-PKA）， and PKA in lung tissues of rats in each group. The level of VIP mRNA in lung tissues of rats was detected by real-time quantitative polymerase chain reaction （Real-time PCR）.ResultCompared with the blank group， the model group exhibited obvious lung injury， increased edema score， decreased D/W ratio （P<0.01）， declined AQP1， AQP5， cAMP， and p-PKA/PKA in lung tissues （P<0.05， P<0.01）， elevated VIP content （P<0.01）， and up-regulated levels of VIP protein and mRNA in lung tissues （P<0.05， P<0.01）. Compared with the model group， combined therapy of lung and intestine treatment groups showed alleviated lung injury， increased D/W ratio （P<0.01）， elevated AQP1， AQP5， VIP， cAMP， and p-PKA/PKA in lung tissues （P<0.05， P<0.01）， and up-regulated VIP levels in lung tissues （P<0.05， P<0.01）.ConclusionThe combined therapy of lung and intestine can alleviate ALI-induced lung tissue edema， and the mechanism may be related to the activation of the VIP/cAMP/PKA signaling pathway， which further promotes the expression of AQP1 and AQP5 and enhances the water metabolism of lung tissue.
Keywords：combined therapy of lung and intestine;acute lung injury;pulmonary edema;vasoactive intestinal peptide （VIP）/cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA）/aquaporins（AQPs） signaling pathway;Mahuangtang;Da Chengqitang
Abstract：ObjectiveTo observe the therapeutic effects of the combined therapy of lung and intestine， a common treatment for pulmonary diseases in traditional Chinese medicine （TCM）， on bronchial asthma mice， and further detect the changes of vasoactive intestinal peptide （VIP） and p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway-related proteins which are closely related to the pathogenesis of asthma， in order to elucidate the mechanism of the combined therapy of lung and intestine in the treatment of bronchial asthma.MethodA total of 60 Kunming mice were randomly divided into normal group， model group， dexamethasone group （0.5 mg·kg-1·d-1）， TCM group （2.73 g·kg-1·d-1）， and lung-intestine treatment group （6.825 g·kg-1·d-1）， 12 mice in each group. All mice except the normal group were sensitized by ovalbumin to induce bronchial asthma. After 30 days of intragastric administration， serum and lung tissue samples were obtained. The content of VIP， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the serum of mice in each group was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA levels of TNF-α， IL-6， and p38 MAPK in lung tissues of mice were detected by real-time quantitative polymerase chain reaction （Real-time PCR）， and the protein levels of TNF-α， IL-6， p38 MAPK， and phosphorylated p38 MAPK （p-p38 MAPK） in lung tissues of mice were assayed by Western blot （WB）.ResultCompared with the normal group， the model group showed decreased content of serum VIP （P<0.05）， increased content of TNF-α and IL-6 （P<0.05）， up-regulated mRNA levels of TNF-α， IL-6， and p38 MAPK， and elevated protein levels of TNF-α， IL-6， and p-p38 MAPK/p38 MAPK in lung tissues （P<0.05）. Compared with the model group， the treatment groups exhibited increased content of serum VIP， TNF-α， and IL-6 （P<0.05）， down-regulated mRNA levels of TNF-α， IL-6， and p38 MAPK， and lower protein levels of TNF-α， IL-6， and p-p38 MAPK/p38 MAPK in lung tissues （P<0.05）. As compared with the lung-intestine treatment group， the serum TNF-α and IL-6 levels in the dexamethasone group were increased （P<0.05）， and the mRNA and protein levels of TNF-α and IL-6 in lung tissues were down-regulated （P<0.05）， while the levels of p38 MAPK， VIP mRNA， and p-p38 MAPK/p38 MAPK protein in lung tissues were up-regulated （P<0.05）. The serum VIP， TNF-α， and IL-6 levels in the TCM group were decreased （P<0.05）， and the mRNA levels of TNF-α， IL-6， p38 MAPK and protein levels of TNF-α， IL-6， p-p38 MAPK/p38 MAPK in lung tissues were up-regulated （P<0.05）， while the level of VIP mRNA in lung tissues was down-regulated （P<0.05）.ConclusionThrough increasing endogenous VIP and inhibiting the excessive activation of p38 MAPK signaling pathway， the combined therapy of lung and intestine can reduce the release of inflammatory factors， inhibit pulmonary inflammation response， and treat bronchial asthma.
Keywords：combined therapy of lung and intestine;vasoactive intestinal peptide;p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway;bronchial asthma;Sanaotang;Xiao Chengqitang
Abstract：The theoretical origin of the combined therapy of lung and intestine can be traced back to the Inner Canon of Huangdi （《黄帝内经》）， which explains the physiological and pathological interaction between the lung and the large intestine. In recent years， researchers have investigated the scientific essence of the "lung-intestine axis" theory from many aspects， which enriches the relevant theoretical basis， and applied it to the treatment of COVID-19， acute lung injury， and other lung diseases. The close relation between lung and intestine in many aspects embodies the holistic conception of traditional Chinese medicine and explains the holistic theory of interrelation between organs， which correlate to each other physiologically and pathologically. Intestinal microecological disorders can affect lung immune function and cause respiratory diseases， and respiratory diseases are usually accompanied by gastrointestinal symptoms. Lung diseases can be prevented and treated by regulating intestinal flora. According to histoembryology， the epithelial tissue of the lung and intestine comes from primitive foregut. In immunology， both lung and intestine contain mucosa-associated lymphoid tissue， and the pathological changes of the respiratory tract are also closely related to intestinal microorganisms. The tissue origin of lung and large intestine， the correlation of mucosal immunity， and the synchronization of ecological changes provide a scientific basis for the combined therapy of lung and intestine. Therefore， this paper summarizes the theoretical origin， modern research mechanism， and clinical application of combined therapy of lung and intestine， in order to provide a new direction for its application in clinical and scientific research.
Keywords：combined therapy of lung and intestine;mucous membrane;microorganism;lung-intestine axis;immunity
Abstract：ObjectiveTo explore the pharmacodynamic ingredients of Zhenqi Fuzheng granules （ZFG） for immunomodulatory through spectrum-effect relationship analysis， which provides experimental basis for improving the quality standard of ZFG.MethodEighteen batches of ZFG from six manufacturers were collected for analysis. The fingerprints were established by high performance liquid chromatography （HPLC）. Acetonitrile （A）-0.1% formic acid aqueous solution （B） were adopted as the mobile phase with gradient elution （0-15 min， 5%A； 15-23 min， 5%-8%A； 23-30 min， 8%-11%A； 30-45 min， 11%-18%A； 45-60 min， 18%-21%A； 60-67 min， 21%-23%A； 67-90 min， 23%-37%A）， the detection wavelength was 220 nm. Chemometric analysis such as similarity analysis and hierarchical cluster analysis （HCA） were subsequently used to analyze the similarities and chemical differences among these samples. A cyclophosphamide-induced immunodeficiency mouse model was used to evaluate the immune-enhancing effects of the products from different manufacturers. The spectrum-effect relationship between HPLC fingerprints and the immunomodulatory effects was examined using Spearman bivariate correlation analysis. HPLC coupled with mass spectrometry （HPLC-MSn） was used to identify the spectrum-effect related peaks with electrospray ionization， positive and negative ion modes， and scanning range of m/z 100-1 500.ResultThe HPLC fingerprint of ZFG was established， and twenty peaks with good resolution were selected as common peaks. The results of quality analysis and pharmacodynamic test showed there were significant differences in both ingredients content and immune-enhancing effects of ZFG from different manufacturers. Through spectrum-effect relationship study， twelve peaks were screened as bioactive ingredients peaks. Thereafter， eight peaks among them were subsequently identified by HPLC-MSn. They were salidroside （peak 2）， echinacoside （peak 5）， calycosin-7-glucoside （peak 6）， isomer of specnuezhenide （peak 7）， isonuezhenide （peak 9）， calycosin （peak 11）， nuezhenide G13 or oleonuezhenide （peak 14）， and formononetin （peak 18）， respectively.ConclusionThere are differences in quality and efficacy of ZFG produced by different manufacturers. Through spectrum-effect relationship analysis， the medicinal ingredients of ZFG for immune-enhancing effects are screened， which can provide reference for the improvement of its quality standard.
Abstract：ObjectiveTo study the effects of foliar spraying of two kinds of compound rhizosphere growth-promoting agents on the growth and physiological characteristics of Angelicae Sinensis Radix （ASR）， as well as the pharmacodynamic components， in order to lay a foundation for providing functional microbial agents for ecological cultivation of ASR.MethodThe compound growth-promoting agents T1 （Pseudomonas CBS5， CBS7 and CBSB） and T2 （Bacillus 5C1， 5C5 and 5C7） with the concentration of 1×108 CFU·mL-1 were sprayed on the leaf surface of the field， and the sterile potato glucose broth medium was used as the control （CK）. The plant growth indexes of ASR were measured by conventional methods， the photosynthetic physiological indexes of ASR were measured by portable photosynthetic measurement system， the enzyme activities of plants and microorganisms were measured by kit method， and the endogenous hormone levels were analyzed by ultra-performance liquid chromatography tandem mass spectrometry. The contents of ferulic acid， senkyunolide I， coniferyl ferulate， senkyunolide A and Z-ligustilide were determined by high performance liquid chromatography.ResultCompared with CK， the two compound inoculants could promote the growth of ASR and increase the biomass， increase the leaf net photosynthetic rate， stomatal conductance， intercellular CO2 concentration， transpiration rate， increase catalase， peroxidase， superoxide dismutase， polyamine oxidase， diamine oxidase and polyphenol oxidase enzyme activities， increase endogenous jasmonic acid， cytokinin and gibberellin levels in plants， increase the contents of ferulic acid， senkyunolide A and Z-ligustilide， reduce the contents of malondialdehyde and abscisic acid， and reduce the incidence of root rot.ConclusionFoliar spraying of two kinds of rhizosphere compound growth-promoting agents can promote the growth， photosynthesis and stress resistance of ASR， and can improve the quality of ASR in different degrees. Comprehensive analysis shows that T1 treatment is better than T2 treatment in the growth-promoting and quality-enhancing of ASR.
Abstract：ObjectiveUltra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS/MS） was used to analyze the chemical constituents in the aerial part and roots of Gentiana straminea from different areas of Qinghai province， and the main chromatographic peaks and differential components of different parts were identified.MethodThe chromatographic separation was performed on a Waters ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 μm） with 0.1% formic acid aqueous solution （A）-acetonitrile （B） as the mobile phase for gradient elution （0-1 min， 1%-13%B； 1-5 min， 13%-18%B； 5-7 min， 18%-50%B； 7-9.5 min， 50%-60%B； 9.5-11 min， 60%-99%B； 11-14 min； 99%B； 14-15 min， 99%-1%B； 15-16 min， 1%B）， the column temperature at 40 ℃， and the flow rate of 0.3 mL·min-1. Electrospray ionization （ESI） and negative ion full scan mode were selected for the mass spectrometric conditions to analyze the samples， and the detection range was m/z 50-1 200. Chemical constituents of the aerial part were qualitatively analyzed with the reference substances， literature information and ChemSpider. Principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to analyze the classification trend， correlation and differential chemical components between aerial part and roots of G. straminea.ResultA total of 68 components， including 24 iridoids， 13 flavonoids， 8 triterpenoids， 6 xanthones， 5 fatty acids， 4 saccharides， 3 phenolic glycosides， 2 alkaloids， 2 sterols and 1 lignan， were preliminarily identified from the aerial part of G. straminea. Among them， 42 components were firstly reported in 4 Gentiana species included in the 2020 edition of Chinese Pharmacopoeia. Eight differential components were screened out， namely sucrose， maltotriose， loganic acid， shanzhiside methyl ester， 6′-O-β-D-glucosylgentiopicroside， swertiamarin， gentiopicrin and isovitexin.ConclusionThe aerial part of G. straminea is rich in chemical constituents and has good medicinal potential. There were significant differences in the chemical components between the aerial part and roots of G. straminea， and the main differential components were iridoids， which could provide a basis for exploring efficacy differences in different parts of G. straminea.
Keywords：Gentiana straminea;chemical composition;ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS/MS）;chemometrics;iridoids;flavonoids;triterpenoids
Abstract：ObjectiveTo compare the effect of different solvent extracts of spore powder and fruiting body of Lasiosphaera Calvatia on fibroblasts and wound healing of full-thickness skin defect， in order to screen the optimal medication part of Lasiosphaera Calvatia.MethodThe effect of water extract and alcohol extract of spore powder and fruiting body on cell proliferation and cell migration of mouse skin fibroblasts （MSF） were examined in vitro. Cell proliferation and activity test （CCK-8） method was used for cell proliferation， scratch assay was used for cell migration， flow cytometry was conducted to explore cell cycle， enzyme-linked immunosorbent assay （ELISA） was used to determine the production of collagen Ⅰ and Ⅲ. At the same time， a full-thickness skin defect wound model was established to investigate the therapeutic effect of different solvent extracts of spore powder. Ultraviolet-visible spectrophotometry was used to measure the contents of index components in different solvent extracts.ResultThe water extract of spore powder and fruiting body had certain cytotoxicity， while the alcohol extract could promote proliferation， migration and production of collagen Ⅰ and Ⅲ of MSF， and the effect of spore powder was significantly higher than that of fruiting body. When the concentration was 10 mg·L-1， the cell proliferation rate of alcohol extract of spore powder was as high as （159.22±15.95）%， and could promote MSF from the G0/G1 phase to S phase and G2/M phase with an increased proliferation index. The alcohol extract also promoted the migration of fibroblasts， secreted collagen Ⅰ and Ⅲ. On in vivo model， the alcohol extract of spore powder significantly accelerated wound healing on mice， effectively promoted the complete epithelialization of wound tissue， and generated new collagen fiber. The results of determination showed that the contents of polyphenols and flavonoids in the alcohol extract were higher than the alcohol extract of fruiting body.ConclusionThe alcohol extract of spore powder in Lasiosphaera Calvatia has active components in the treatment of wounds with good development prospect， and the medicinal components may be polyphenols and flavonoids.
Abstract：ObjectiveTo study the effect of flower removal on the content of three alkaloids in different parts of Fritillaria thunbergii from different regions and at different growth stages.MethodThe content of peiminine， peimine， and peimisine in the bulb， root， stem， and leaf of F. thunbergii after flower removal and with flower un-removed at different growth stages and in different regions were determined simultaneously by ultra-performance liquid chromatography-evaporative light scattering detection （UPLC-ELSD） method. The UPLC was conducted on ACQUITY UPLC BEH C18 column （2.1 mm × 150 mm， 1.7 μm） with the mobile phase of 0.02% triethylamine aqueous solution （A） and methanol （B）elution gradient（0-2 min， 45%A； 2-5 min， 45%-25%A； 5-7 min， 25%A； 7-17 min， 25%-10%A； 17-20 min， 10%A）， flow velocity of 0.20 mL·min-1， column temperature 35 °C， sample room temperature of 20 °C， and injection volume of 3 µL. The ELSD was carried out at drift tube temperature 45 °C and with the sprayer parameter of 40%.ResultThe flower removal significantly increased the yield of F. thunbergii. At the budding stage， the alkaloid content in the bulb of F. thunbergii from Ningbo in Zhejiang， Pan'an in Zhejiang， and Nantong in Jiangsu after flower removal were significantly higher than that of flowering un-removal treatment， while it showed no significant difference between the flower removal and un-removal treatments for the samples from Fengjie in Chongqing. At the flowering stage， the alkaloid content in the bulb of F. thunbergii from Nantong in Jiangsu after flower removal was significantly higher than that of flower un-removal treatment， while it showed an opposite trend for the samples from Pan'an in Zhejiang and Fengjie in Chongqing and had no significant difference between the two treatments for the samples from Ningbo in Zhejiang. At the bulb expansion stage， the alkaloid content in the bulb of F. thunbergii from Ningbo in Zhejiang and Pan’an in Zhejiang after flower removal were significantly higher than that of flower un-removal treatment， which was opposite for the samples from Nantong in Jiangsu and had no significant difference between the treatments for the samples from Fengjie in Chongqing. At the harvest stage， except for the samples from Pan'an in Zhejiang， the samples from the rest 3 regions showed decreased alkaloid content in the bulb after flower removal compared with that of flower un-removal treatment. The alkaloid content in the leaf was higher than that in the bulb of F. thunbergii at all growth stages and from different origins.ConclusionFlower removal can increase the yield of F. thunbergii. The alkaloid content in the bulb of F. thunbergii with flower removed was higher than that with flower un-removed at the budding stage， while this trend was reversed at the harvest stage. Both the yield and the alkaloid content of F. thunbergii from Pan'an in Zhejiang were increased by flower removal. The above-ground part of F. thunbergii has a potential development value.
Abstract：ObjectiveTo predict the active ingredients and mechanism of action of lavender in protecting skin photodamage based on network pharmacology and molecular docking technology，and further verify possible signal pathways via animal experiments.MethodThe active ingredients and potential targets of lavender were obtained by SwissTargetPrediction，PharmMapper， and literature. Skin photodamage-related targets were searched from GeneCards，Online Mendelian Inheritance in Man （OMIM），DrugBank and DisGeNET databases. After common targets of the two were screened out，STRING was adopted to analyze the protein-protein interaction （PPI） network，where topological analysis and core target screening were performed by CytoNCA plug-in of Cytoscape 3.8.2. Based on DAVID， gene ontology （GO） annotations and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were carried out among the intersection targets， and the active ingredients of lavender and the signal pathway proteins were selected and verified via molecular docking with AutoDock vina 1.1.2. Finally， mouse photodamage model was established by UVB irradiating the bare skin of mouse back， and the skin condition was observed by naked eyes. Hematoxylin-eosin （HE） and picric acid-acid fuchsin staining （Van Gieson， VG） were used to observe the pathological changes of mouse skin tissues. Western blot was employed to detect the protein expression in mouse skin tissues to further validate the key signal pathways.ResultIn this study，6 active ingredients of lavender，526 potential targets，2 688 disease-related targets，and 258 intersection targets were screened out， and 16 core targets were obtained by PPI network. Additionally， 113 related signal pathways were obtained by KEGG pathway analysis，among which phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathway and nuclear transcription factor-κB （NF-κB） signal pathway might play a key role in skin photodamage protection by lavender. Molecular docking showed that the active ingredients and the signal pathway proteins were well docked. Animal experiments indicated that the total flavonoids of lavender improved the appearance and histopathological condition of mouse skin， reduced the relative expression levels of phosphorylated（p）-PI3K，p-Akt，and B cell lymphoma 2 （Bcl-2） proteins （P<0.05，P<0.01）， and increased relative expression level of Bcl-2-associated X protein（Bax） （P<0.05）.ConclusionLavender exerts synergistic effect in resisting skin photodamage，with the characteristics of multi-components，multi-targets，and multi-pathways， which provides a basis for subsequent in-depth research on the complex mechanism of lavender against skin photodamage.
Keywords：lavender;skin photodamage;network pharmacology;molecular docking;phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signal pathway
Abstract：ObjectiveTo explore the mechanism of Fangji Fulingtang in the treatment of acute kidney injury （AKI） induced by ischemia-reperfusion based on network pharmacology and experimental verification.MethodActive components of Fangji Fulingtang were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and previous report and targets of these components were predicted by SwissTargetPrediction. The targets of AKI were searched from GeneCards， Online Mendelian Inheritance in Man （OMIM）， the database of gene-disease associations （DisGeNET）， and Therapeutic Target Database （TTD）. Protein-protein interaction （PPI） network was constructed by STRING. Metascape was used for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of core targets. Cytoscape was employed to construct the "medicinal-active component-target-disease" network and “active component-target-pathway” network. AutoDock was applied for molecular docking. Finally， animal experiment was carried out to validate the mechanism of Fangji Fulingtang in treatment of AKI.ResultA total of 137 active components and 858 targets of Fangji Fulingtang， 1 294 targets of AKI， and 267 targets of Fangji Fulingtang in the treatment of AKI were screened out. Phosphoinositide-3-kinase regulatory subunit 1 （PIK3R1）， phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit alpha （PIK3CA）， proto-oncogene tyrosine protein kinase （SRC）， protein kinase B1 （Akt1）， and mitogen-activated protein kinase 3 （MAPK3） were the key anti-AKI targets of Fangji Fulingtang， which were involved in 1 609 GO terms， particularly cell response to lipids， membrane rafts， and protein kinase activity， and 140 KEGG pathways such as PI3K/Akt signaling pathway， chemokine signaling pathway， and Toll-like receptor signaling pathway. Molecular docking showed that the core active components had strong binding affinity to the key targets. The hematoxylin and eosin （HE） staining results indicated that Fangji Fulingtang can significantly improve the pathological state and the serological results suggested that the levels of serum creatinine （SCr） and blood urea nitrogen （BUN） were significantly reduced.ConclusionThis study clarified the mechanism of Fangji Fulingtang in the treatment of AKI and found that Fangji Fulingtang had the multi-component， multi-target， and multi-pathway characteristics in the treatment of AKI. The result lays a foundation for further study of its specific mechanism.
Abstract：ObjectiveTo systematically analyze the chemical components of QiLing Wenshen （QLWS） formula and explore the key active components and mechanism of the formula in the treatment of polycystic ovary syndrome （PCOS）.MethodThe chemical components of QLWS formula were systematically identified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE） combined with comparison with reference substances， literature data， and databases. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and SwissADME were employed to screen the active components for network pharmacological analysis. SwissTargetPrediction， GeneCards， DisGeNET， and DrugBank were used to obtain the potential components and targets of the formula for the treatment of PCOS. The protein-protein interaction （PPI） network was constructed via STRING database for further screening of the core targets. Gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of core targets were carried out with DAVID database. Molecular docking was performed in MOE 2019.ResultA total of 90 components of QLWS formula were identified， and 32 active components and 45 core targets for treating PCOS were obtained. GO annotation obtained 429 terms and KEGG pathway enrichment screened out 110 signaling pathways， mainly involving phosphatidylin-ositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， estrogen signaling pathway， and hypoxia inducible factor-1 （HIF-1） signaling pathway. The molecular docking revealed that key active components in QLWS formula were icariin， salvianolic acid A\B\C， wogonin， magnoflorine， etc.， which may play a role in treating PCOS through regulating mitogen-activated protein kinase 1 （MAPK1）， epidermal growth factor receptor （EGFR）， mitogen-activated protein kinase 3 （MAPK3）， etc.ConclusionThis study preliminarily predicted that several key active components of QLWS formula could treat PCOS via multiple targets and multiple pathways based on UPLC-Q-TOF/MSE and network pharmacology， which could provide ideas and references for the study of pharmacodynamic material basis and mechanism of action of the formula.
Keywords：QiLing Wenshen formula;polycystic ovary syndrome;network pharmacology;ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry;molecular docking;mechanism of action
Abstract：ObjectiveTo reveal the medication rules of the clinical prescriptions containing Citri Sarcodactylis Fructus， and to provide a basis for the modern clinical application of Citri Sarcodactylis Fructus， the development of health products and the research and development of new drugs.MethodThe clinical prescriptions containing Citri Sarcodactylis Fructus were retrieved from CNKI， Wanfang Data， and VIP， and then a basic database of prescriptions was established via the traditional Chinese medicine inheritance auxiliary platform （V2.5）， IBM SPSS V20， and Excel （Microsoft Office 2016）. The frequency and association rules of the medicines in prescriptions （compatible medicines） and the corresponding syndromes were then mined.ResultThe prescriptions were screened according to the inclusion and exclusion criteria. A total of 458 clinical prescriptions containing Citri Sarcodactylis Fructus were collected， involving 388 Chinese medicines， and the total frequency of medicines reached 6 199. The core compatible medicines （frequency > 130） of Citri Sarcodactylis Fructus included Poria （frequency of 222）， Atractylodis Macrocephalae Rhizoma （217）， Paeoniae Radix Alba （196）， Bupleuri Radix （159）， and Citri Reticulatae Pericarpium （142）. The Citri Sarcodactylis Fructus-compatible medicines with frequency > 49 were selected for further analysis， which included 34 medicines with the cumulative frequency of 3 131 （50.51% of the total frequency）. These medicines mainly have the functions of tonifying Qi， invigorating Qi， tonifying blood， alleviating edema and promoting urination， promoting digestion， and activating blood and relieving pain. They are mainly warm， cold， or mild-natured， taste bitter， sweet， or acrid， and have the tropism in the spleen， liver， stomach， or lung meridians. The association rule analysis demonstrated that 14 medicine combinations were commonly used， and the core combinations were Poria-Citri Sarcodactylis Fructus， Atractylodis Macrocephalae Rhizoma-Citri Sarcodactylis Fructus， Paeoniae Radix Alba-Citri Sarcodactylis Fructus， Bupleuri Radix-Citri Sarcodactylis Fructus， Atractylodis Macrocephalae Rhizoma-Poria-Citri Sarcodactylis Fructus， and Citri Reticulatae Pericarpium-Citri Sarcodactylis Fructus. The clinical prescriptions containing Citri Sarcodactylis Fructus were mainly used to treat 52 diseases corresponding to 11 types of traditional Chinese medicine （TCM） syndromes. Three representative syndrome types， including spleen and stomach syndromes， Qi-blood-body fluid syndromes， and gynecological syndromes were selected for further association rule analysis. In the treatment of spleen and stomach syndromes， the core compatible drugs were Atractylodis Macrocephalae Rhizoma， Poria， Paeoniae Radix Alba， Bupleuri Radix， Citri Reticulatae Pericarpium， and Pinelliae Ehizoma， which， together with Citri Sarcodactylis Fructus， formed 25 commonly used medicine combinations （16 combinations composed of 2 medicines and 9 combinations composed of 3 medicines）. In the treatment of Qi-blood-body fluid syndromes， the core compatible drugs were Poria， Paeoniae Radix Alba， Atractylodis Macrocephalae Rhizoma， Astragali Radix， Hordei Fructus Germinatus， and Bupleuri Radix， which， together with Citri Sarcodactylis Fructus， formed 23 common medicine combinations （17 combinations composed of 2 medicines， 5 combinations composed of 3 medicines， and 1 combination composed of 4 medicines）. In the treatment of gynecological syndromes， the core compatible medicines were Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， Angelicae Sinensis Radix， Astragali Radix， Cyperi Rhizoma， and Poria， which constituted 25 common medicine combinations （15 combinations composed of 2 medicines and 10 combinations composed of 3 medicines）.ConclusionWe employed the traditional Chinese medicine（TCM） inheritance auxiliary platform to explore the compatibility of Citri Sarcodactylis Fructus-containing clinical prescriptions and the corresponding TCM syndromes， which intuitively showcased the medication rules of Citri Sarcodactylis Fructus. Specifically， Citri Sarcodactylis Fructus was mainly combined with the medicines for tonifying Qi， invigorating Qi， tonifying blood， alleviating edema and promoting urination， promoting digestion， and activating blood and relieving pain to treat different TCM syndromes. While soothing liver， regulating Qi， harmonizing stomach， and relieving pain， the combinations tonify and activate blood， invigorate spleen， and resolve dampness. The findings are of great significance to the rational application of Citri Sarcodactylis Fructus， the development of health food， and the research of new drugs and will bolster the development of Chinese medicine industry.
Keywords：Citri Sarcodactylis Fructus;data mining;clinical prescriptions;medication rule;traditional Chinese medicine inheritance auxiliary platform
Abstract：This scoping review aimed to summarize the application information and clinical studies of oral Chinese patent medicines. The oral Chinese patent medicines in treating lung cancer were screened out by searching pf the drug directory， related guidelines， and medical information websites. The data including functions， application， ingredients， and prices of these medicines were collected. Six public databases were searched with the time interval of establishment to August 22， 2021 for collection of the clinical studies of oral Chinese patent medicines in the treatment of lung cancer. The expert consensuses， systematic reviews， randomized controlled trials， non-randomized controlled trials， and non-controlled trials were selected for analysis. A total of 104 oral Chinese patent medicines were screened out， including 31 capsules， 16 granules， 20 oral liquids， 17 tablets， 17 pills， and 3 ointments， in which altogether 198 herbal medicines were involved. The single-dose prices of 2， 36， and 66 medicines were > CNY 100， CNY 10-100， and < CNY 10， respectively. There were 410 clinical studies associated with 48 oral Chinese patent medicines， which were published from 1986 to 2021. These publications included 1 expert consensus， 21 systematic reviews， 277 randomized controlled trials， 87 non-randomized controlled trials， and 24 non-controlled trials. In the clinical studies， the Chinese patent medicines were usually applied in combination with radiotherapy and chemotherapy. The evaluation of primary outcomes focused on 9 indicators including clinical efficacy， quality of life， and incidence of side effects. In conclusion， the oral Chinese patent medicines demonstrated significant advantages in the treatment of lung cancer， and the relevant clinical trials were increasing year by year， with multiple outcome indicators being evaluated. More comprehensive and standardized clinical studies need to be designed for oral Chinese patent medicines in treating lung cancer in the future.
Keywords：oral Chinese patent medicine;lung cancer;clinical research;scoping review
Abstract：To summarize the status quo of clinical evidence on oral Chinese patent medicine in the treatment of hyperlipidemia through scoping review and thereby provide a reference for clinical application and decision-making in health care. Proprietary Chinese medicines for the treatment of hyperlipidemia （dyslipidemia） were retrieved from relevant catalogs and then screened based on their instructions. Articles on the selected Chinese patent medicines were searched from Chinese and english electronic databases and screened according to the inclusion criteria， followed by data extraction and analysis. The results were described with text and graphs. ①A total of 32 Chinese patent medicines and 1 010 related articles were screened out. ②All the included Chinese patent medicines were made from Chinese medicinal materials， of which 5 were Chinese medicinal extracts，and the remaining 27 were pure Chinese medicinal preparations （the compositions of 1 prescription is confidential）. ③As indicated in the instructions， all the 32 Chinese patent medicines can be used to treat hyperlipidemia with the main syndrome of combined phlegm and blood stasis， and the main effect of them is activating blood and resolving stasis. ④Among the 32 medicines， Xuezhikang tablets （Capsules） and Gypenosides Tablets have the lowest price. ⑤For the 328 key clinical studies， 248 were randomized controlled trials and 80 non-randomized controlled trials. ⑥As for the sample size， randomized controlled trials generally included 28-579 cases， and non-randomized controlled trials 24-152 cases. Clinical studies with more than 200 cases accounted for 9.3% （12/129）. ⑦The most common method was the comparison of the intervention effect of Chinese patent medicine and western medicine （36.5%） and Atorvastatin Calcium Tablets was the preferred western medicine control. The observation duration was generally 56 days （8 weeks）. ⑧In clinical practice， the absolute value of blood lipid decrease and （total） effective rate were often used to evaluate the efficacy， and the effective rate was generally determined based on Clinical Guidelines for New Chinese Medicines. The overall quality of clinical studies on oral Chinese patent medicines is uneven and there is a lack of high-quality clinical evidence. Moreover，there are the risks of unreasonable use and uneasy use of Chinese patent medicine.It is recommended that researchers on proprietary Chinese medicine should further broaden the research ideas，focus on the top-level design of the research plan， and standardize the research process，thereby provide high-quality research evidence for the clinical use of proprietary Chinese medicine. Moreover， more efforts should be made to establish a sound mechanism for information collection and feedback of proprietary Chinese medicine， so as to reduce the risk of uneasy use.We hope that in-depth investigation and discussion should be launched by price， economy， quality evaluation， and other departments headed by the supervision department to make the price of Chinese patent medicine reasonable and fair， thereby promoting the rational use of Chinese patent medicine.
Keywords：hyperlipidemia;dyslipidemia;Chinese patent medicine;clinical research;scoping review;rational use of drugs
Abstract：This study aims to summarize the clinical research evidence on oral Chinese patent medicines for the treatment of influenza with the method of scoping review， and thus clarify the status quo and problems. Specifically， the target medicines were selected from related drug catalogues and diagnosis and treatment protocols， and the basic information of the medicines on the specifications was collected. Articles on these medicines were retrieved from Chinese and English databases for statistical analysis and visualization. Finally， 36 medicines and 87 articles were included. The main efficacy of the medicines is clearing heat and removing toxin， and the main components of the medicines are Lonicerae Japonicae Flos， Forsythiae Fructus， and Isatidis Radix. A total of 12 medicines can be used for the treatment of mumps and acute bronchitis in addition to influenza. Only 6 medicines have contraindications and adverse reactions labeled on the specifications. Papers on oral Chinese patent medicines in the treatment of influenza show an increasing trend， and the authors are from 25 provinces and cities in China. Among them， papers on Lianhuaqingwen preparations take up the largest proportion. The studies were mostly randomized controlled trials， non-randomized controlled trials， and retrospective research. A total of 13 studies were supported by national funding， and only 9 studies included more than 200 cases. The most frequently used method was the comparison of the intervention effect of Chinese patent medicines with western medicine， and the treatment course was generally 3-14 days. A total of 7 outcome indicators were used in the studies and the frequency was in the order of ① composite effective rate，② antipyretic effect， ③ symptom improvement， ④ safety indicator， ⑤ virological examination， ⑥ serum inflammatory factor， and ⑦ traditional Chinese medicine （TCM） syndrome score. The conclusions in the clinical studies show difference from the information in drug catalogues. The drug specifications are generally not standard. The available clinical studies have the limitations of small quantity， low in quality， and no demonstration of TCM advantages. In the future， it is necessary to optimize the specifications of Chinese patent medicines， enhance clinical research， further standardize the design of clinical research， and highlight the characteristics of Chinese patent medicines， thereby providing evidence to support the comprehensive clinical evaluation of oral Chinese patent medicines for influenza.
Keywords：oral Chinese patent medicines;influenza;clinical research;scoping review
Abstract：To clarify the research progress and existing problems in the clinical research on oral Chinese patent medicine in the treatment of breast cancer. National drug catalogs and databases were searched for oral Chinese patent medicines against breast cancer. Chinese and English databases were searched for clinical trials on oral Chinese patent medicines in the treatment of breast cancer. The main functions，drug composition，literature basic characteristics，intervention measures， and outcome indicators of Chinese patent medicines were comprehensively analyzed and evaluated by a scoping review based on visual charts. In 68 Chinese patent medicines included，53% of them were potent in activating blood， resolving stasis， tonifying the kidney， and strengthening the spleen. In terms of frequency， the top drugs were Ligustri Lucidi Fructus， Astragali Radix， Moutan Cortex， Poria， and Olibanum，indicating the importance of the therapeutic principles of activating blood， resolving stasis， tonifying the kidney， and strengthening the spleen for breast cancer treatment. In 161 included clinical trials，70.71% of the intervention measures were Chinese patent medicine combined with chemotherapy. Literature studies mainly reported Xihuang pills and Huaier granules，which may be related to the combination of chemotherapy and the inclusion in the medical insurance. The outcome evaluation was mainly based on immune indexes，tumor markers，Karnofsky score，safety report and objective response rate，while the survival time and characteristic outcome indicators of traditional Chinese medicine （TCM） were insufficient. The application and research on oral Chinese patent medicine in the treatment of breast cancer have advanced in recent years，but most of the clinical research evidence is not robust，and the evaluation of outcome indicators is not standardized，which needs to be improved.
Keywords：breast cancer;oral Chinese patent medicine;scoping review;clinical research;rational medication of drugs
Abstract：This study systematically evaluated the effectiveness and safety of Dingkundan combined with conventional western medicine in the treatment of female infertility. Four Chinese databases，three English databases， and two clinical trial registration platforms were retrieved from inception to April 2021. Two researchers independently carried out literature screening，data extraction，risk assessment of bias，and grading of evidence quality. RevMan 5.4.1 was used for data analysis. A total of 216 research articles were retrieved and 21 randomized controlled trials （RCTs） were included，involving 2 172 cases. The risks of bias in the included RCTs were high. As unraveled by Meta-analysis results， Dingkundan combined with western medicine for ovulation stimulation was superior to western medicine for ovulation stimulation alone in improving pregnancy rate and progesterone level ［relative risk（RR）pregnancy rate=1.67，95% confidence interval（CI）（1.44，1.93）；standardized mean difference （SMD）progesterone=1.21，95% CI（0.82，1.60）］. Dingkundan combined with western medicine for improving the endometrium was superior to western medicine for improving the endometrium alone in improving the pregnancy rate ［RRpregnancy rate=1.35，95% CI（1.23，1.48）］. Dingkundan combined with clomiphene was more effective than clomiphene alone in regulating endometrial thickness and reducing follicle-stimulating hormone and estradiol levels ［MDendometrial thickness=3.34，95% CI（3.27，3.41）， MDfollicle-stimulating hormone=-0.42，95% CI（-0.65，-0.19）， MDestradiol=-4.33，95% CI （-8.18，-0.48）］. Dingkundan combined with letrozole was superior to letrozole alone in increasing the follicle-stimulating hormone level and reducing the estradiol level ［MD follicle-stimulating hormone=1.14，95% CI（0.49，1.78）， MDestradiol =-33.65，95% CI（-59.13，-8.17）］. The single-study results showed that Dingkundan combined with conventional western medicine had certain advantages in regulating endometrial thickness，reducing follicle-stimulating hormone，luteinizing hormone，and estradiol levels，and increasing progesterone levels. The Grading of Recommendations， Assessment， Development， and Evaluation（GRADE）system was used for the evaluation of outcome indicators. The results showed that the quality of the evidence was graded moderate or low. Based on the existing evidence，Dingkundan combined with western medicine for infertility treatment had certain advantages in increasing the pregnancy rate， improving endometrial thickness， regulating hormone levels， and reducing adverse reactions. However，affected by the quality of the included trials，the results may have limitations，and high-quality RCTs are needed for verification in the future.
Abstract：Alzheimer's disease （AD） is a neurodegenerative disease. With the acceleration of aging process， the number of AD patients increases year by year. This threatens the health and even life of patients， and causes heavy economic burden and mental pressure to patients， families and society. In traditional Chinese medicine （TCM）， AD belongs to the category of dementia， and tonifying kidney is the main treatment. Based on the basic theory of TCM and combined with clinical manifestations of AD， AD is closely correlated with liver and spleen. Therefore， "simultaneous regulation of three Yin" of liver， spleen and kidney will be an important way for the prevention and treatment of AD. Hei Xiaoyaosan， a representative prescription of "simultaneous regulation of three Yin" of liver， spleen and kidney， has theoretical， experimental and clinical basis in preventing and treating AD. Modern studies have shown that neurofibrillary tangle formed by tau hyperphosphorylation is a main pathological feature of AD， and trimethylamine oxide （TMAO） is closely related to tau hyperphosphorylation. Therefore， regulating TMAO metabolism to inhibit tau hyperphosphorylation is a new target for the prevention and treatment of AD. On the basis of the above theory and previous studies， this paper put forward the hypothesis that Hei Xiaoyaosan regulates the trimethylamine（TMA）/heparin monooxygenase 3（FMO3）/TMAO metabolic pathway of intestinal flora through "simultaneous regulation of three Yin" of liver， spleen and kidney， and then inhibits tau hyperphosphorylation in brain hippocampus， thereby protecting nerve cells， improving learning and memory， and preventing AD. This paper explored the role and mechanism of Hei Xiaoyaosan in the prevention and treatment of AD from the perspective of inhibiting tau hyperphosphorylation by regulating the TMA/FMO3/TMAO metabolic pathway of intestinal flora， which provided new ideas and strategies for in-depth study of Hei Xiaoyaosan in the prevention and treatment of AD.
Keywords：Alzheimer's disease （AD）;"simultaneous regulation of three Yin "of liver， spleen and kidney;Hei Xiaoyaosan;intestinal flora;tau hyperphosphorylation
Abstract：With the development of omics technology， the construction of disease networks has been widely used in the study of complex diseases. It has been widely used to construct disease networks using systems biology technology to study complex diseases. The mechanism exploration model of disease molecular network which uses the method of constructing disease networks， simulates the occurrence of diseases， explores the core development mechanism of complex diseases， and then predicts biomarkers and exploits the mechanism of drug action provided many new thoughts for the prevention and treatment of complex diseases. Nowadays， the research on the mechanism of myocardial infarction caused by myocardial ischemia and heart failure after myocardial infarction is still very important. However， the research of the molecular network of myocardial infarction and heart failure diseases is usually limited to a few targets and pathways， so it is not able to comprehensively and systematically explain the disease process. Furthermore， authors outlined the typical biological process of "myocardial infarction-heart failure" and related targets from the pathophysiological level， and summarized the existing methods of constructing dynamic networks for heart diseases and other diseases. Based on the dynamic molecular network construction methods of cardiac diseases and other diseases， this paper discusses the construction of the dynamic molecular network of myocardial infarction and heart failure， in order to understand the evolution of myocardial infarction and heart failure more accurately and explore the importance of the dynamic molecular network of the disease process for the study of disease mechanism.
Abstract：Diabetic nephropathy （DN） is classified into "collateral diseases" in traditional Chinese medicine （TCM）. This study reviewed， analyzed， and discussed the clinical and basic research on DN treatment from collateral diseases in recent years. The main TCM therapeutic principles of DN from collateral diseases are dominated by reinforcing the deficiency and dredging collaterals. Fundamental research showed that the relevant mechanism is related to the improvement of oxidative stress and endoplasmic reticulum stress， reduction of inflammatory responses， coping with microcirculation disorder and vascular endothelial damage， and alleviation of renal injury. The theory of collateral diseases can provide ideas for clinical and basic research on DN and also shows advantages in clinical application. However， the present clinical application of this therapeutic principle is mainly based on the combination of deficiency-reinforcing drugs and blood stasis-resolving drugs. In spite of some overlap， blood stasis syndrome and collateral diseases are not in the same category. Therefore， the current diagnosis and treatment of DN from collateral diseases fail to combine with the changes of collateral vessels， main pathogenic factors， and the changes of the secondary pathogenesis. The application of classic collateral drugs such as pungent drugs， rattan drugs， and insect drugs has not been paid enough attention in syndrome differentiation and treatment. In addition， there is a lack of correlation with clinical symptoms and laboratory indexes in the basic research on the treatment of DN by Chinese medicinal prescriptions. Accordingly， the present study advocates systematic research on DN under the guidance of collateral diseases theory， clarifying the essence of DN in collateral diseases and emphasizing the importance of collateral drugs and microscopic syndrome differentiation in screening target drugs for DN to explore the underlying mechanism of collateral drugs in the treatment of DN， so as to provide evidence for the clinical application of collateral diseases theory and Chinese medicinal prescriptions against collateral diseases in the treatment of DN.
Abstract：The occurrence and development of colorectal cancer as a complex disease involves the abnormality of multiple signaling pathways. Chinese medicine regulates a variety of biological processes such as tumor cell differentiation， cell proliferation， apoptosis， cell metastasis， cell cycle， and tumor angiogenesis to prevent the occurrence of colorectal cancer （inflammation-cancer transformation）， tumor metastasis （common metastases of colorectal cancer include liver metastasis， lung metastasis， bone metastasis， and lymphatic metastasis）， and multidrug resistance induced by chemotherapy， treat primary tumors， and mitigate the toxic and side effects of chemotherapy. The pathways of Chinese medicine in the treatment of colorectal cancer have been intensively studied. The available studies have demonstrated that Patrinia villosa Juss and Pien Tze Huang can regulate the Notch pathway to inhibit the growth of colorectal cancer cells. Curcumin and Quyu Jiedu decoction regulate Hippo pathway to inhibit the survival， proliferation， invasion， and migration of colorectal cancer cells. Kujin tea and luteolin suppress the proliferation of colorectal cancer cells and protect intestinal barrier by regulating Kelch-like epichlorohydrin-associated protein-1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/antioxidant response element （ARE） pathway. Icariin and ginkgolide C can regulate hepatocyte growth factor （HGF）/cellular-mesenchymal to epithelial transition factor （c-Met） pathway to induce apoptosis of colorectal cancer cells and prevent liver metastasis of colorectal cancer. Verbascoside and apigenin regulate p53 protein to promote apoptosis of colorectal cancer cells， reverse thymidylate synthase （TS）， and alleviate the multidrug resistance of colorectal cancer. Resveratrol and lycopene regulate insulin-like growth factor （IGF）/insulin-like growth factor receptor 1 （IGF1R） pathway to inhibit cancer cell metastasis and prolong disease-free survival. Cordycepin and Galla Chinensis water extract activate AMP-activated protein kinase （AMPK） pathway to inhibit the migration and invasion of cancer cells as well as the lung metastasis of colorectal cancer. The above summary aims to provide reference for the in-depth research on the treatment of colorectal cancer with Chinese medicine and inspire new research ideas.