Abstract：ObjectiveTo explore the effect of Guilu Erxianjiao on improving reproductive injury in diabetic rats and its possible mechanism.MethodFifty-three SD male rats were randomly divided into 5 groups， with 1 group as the normal group and the other 4 groups as the modeling groups. Rats in the modeling groups were fed with a high-fat diet combined with 30 mg·kg-1 streptozotocin （STZ） intraperitoneal injection to induce diabetes， with the random blood glucose >16.7 mmol·L-1 for 3 consecutive times as the criteria for inclusion in the model of diabetic reproductive injury. The rats with diabetic reproductive injury were then randomly divided into a model group， a Guilu Erxianjiao group （2 g·kg-1）， a Vitamin E group （0.03 g·kg-1）， and a Wuzi Yanzong pill group （0.6 g·kg-1） according to the blood glucose level. The rats were given the corresponding drug dose intragastric administration， once a day for 4 weeks， and their body weight and blood glucose were measured weekly. After 4 weeks， samples were collected for index determination. Morphological changes in testis and epididymis were observed by hematoxylin-eosin （HE） staining， and apoptosis of testis cells was observed by in situ end labeling （TUNEL） staining. Sperm concentration and motility were detected by the semen analyzer. Serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） were determined by enzyme-linked immunosorbent assay （ELISA）. The content of superoxide dismutase （SOD）， propylene glycol （MDA）， reactive oxygen species （ROS）， and glutathione peroxidase （GSH-Px） in the testicular tissue was determined by ELISA. The expressions of nuclear respiratory factor-2 （Nrf2）， heme oxygenase 1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in the testicular tissue were detected by Western blot.ResultAs compared with the normal group， testicular tissue atrophy， decreased spermatogenic tubules， epididymal wall hyperplasia， and lumen stenosis were observed in the model group. Sperm concentration and motility decreased （P<0.01）， and serum levels of T， FSH， and LH decreased （P<0.01） in the model group. The content of ROS and MDA in the testis increased （P<0.01）， while that of SOD and GSH-Px decreased （P<0.01） in the model group. The expression of Bax increased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 decreased （P<0.01） in the model group. As compared with the model group， the pathological changes in the testis and epididymis in the Guilu Erxianjiao group were improved to some extent. Sperm concentration and motility increased （P<0.05， P<0.01）. In the Guilu Erxianjiao group， serum levels of T and LH increased （P<0.05， P<0.01）， while FSH levels showed no significant difference. The content of ROS and MDA in the testis decreased （P<0.01）， while that of SOD and GSH-Px increased （P<0.01） in the Guilu Erxianjiao group. The expression of Bax decreased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01） in the Guilu Erxianjiao group.ConclusionGuilu Erxianjiao improves the reproductive injury and sperm quality of diabetic rats to a certain extent， and the mechanism may be related to the improvement of oxidative stress and anti-apoptosis.
Abstract：ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages.MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl2）. The cells were randomly divided into blank group， CoCl2 hypoxia model group （200 mmol·L-1）， Biejiajian Wan low-dose group （200 mmol·L-1+0.55 g·kg-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L-1+1.1 g·kg-1 Biejiajian Wan）， and high-dose group （200 mmol·L-1+2.2 g·kg-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L-1+0.56 g·kg-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence.ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl2 stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited.ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.
Keywords：Biejiajian Wan;macrophages;hypoxia inducible factor-1α （HIF-1α）;nuclear factor-kappa B （NF-κB）;inflammatory factors
Abstract：ObjectiveTo investigate the preventive and curative effect of Chaishao Liujunzi Tang （CSLJZT） on colonic mucosal injury induced by dextran sulfate sodium （DSS） in mice with ulcerative colitis （UC） and its mechanism.MethodFifty Balb/c male mice were randomly divided into normal group， model group， CSLJZT low-dose group， CSLJZT high-dose group， and sulfasalazine group. Except for the normal group， other groups were given 2.5% DSS freely for 7 d， and were given drug intervention after successful modeling for 7 d. Bodyweight， feces， and other general physiological statuses of mice were recorded every day， and disease activity index （DAI） scores were calculated.The colon length was measured， and stained by hematoxylin-eosin （HE） staining to observe the morphological changes of the colon.The enzyme-linked immunosorbent assay （ELISA） was used to determine the content of interleukin-1β （IL-1β）， myeloperoxidase （MPO）， and superoxide dismutase （SOD） in the serum. Western blot was used to determine the protein expression levels of Toll-like receptor 4 （TLR4）， myeloid differentiation factor 88 （MyD88）， nuclear factor kappa B （NF-κB）， inhibitor-kappa binding protein （IκB）， Caspase-1， and nucleotide-binding oligomerization domain-like receptor 3 （NLRP3） in the colon tissues.ResultAs compared with the normal group， mice in the model group had significantly decreased body weight （P<0.01）， severe diarrhea and hematochezia， and significantly increased DAI score （P<0.01）. As compared with the model group， the decreasing trend of body weight was significantly alleviated in the CSLJZT groups （P<0.01）， diarrhea and hematochezia were significantly improved， DAI score was significantly decreased （P<0.01）， and colon length increased （P<0.05）. HE staining showed that the pathological damage of colon tissue was significantly improved and the inflammatory cell infiltration was reduced in the CSLJZT groups as compared with the model group. As compared with the normal group， the serum levels of IL-1β and MPO were significantly higher （P<0.01） and SOD levels were significantly lower （P<0.01） of mice in the model group.Compared with the model group， the treated group reduced the serum IL-1β and MPO levels （P<0.01）， and raised the SOD level （P<0.01）. The results of Western blot showed that as compared with the normal group， the expression levels of TLR4， MyD88， NF-κB， Ccaspase-1， and NLRP3 proteins were significantly increased （P<0.01）， whereas the expression level of IκB protein was significantly decreased （P<0.01） in the colonic tissue of mice in the model group. As compared with the model group， the expression levels of TLR4， MyD88， NF-κB， Caspase-1， and NLRP3 proteins were decreased （P<0.01）， whereas the expression level of IκB protein was increased （P<0.01） in the colonic tissue of mice in the CSLJZT groups.ConclusionCSLJZT improves the inflammatory injury of the colon tissue in DSS-induced UC mice through TLR4/MyD88/NF-κB signaling pathway.
Abstract：ObjectiveTo explore the protective effect of Salviae Miltiorrhizae Radix et Rhizoma and Puerariae Lobatae Radix （SP） extract on oxygen-glucose deprivation/reoxygenation （OGD/R）-injured SH-SY5Y cells based on oxidative stress and apoptosis.MethodThe extracts of the two medicinal materials mixed in different ratios were prepared. Human neuroblastoma SH-SY5Y cells were cultured in vitro and the injury was induced by OGD/R. Cell counting kit-8 （CCK-8） assay was used to screen the optimal ratio of the two medicinals and then the extract was used for further experiment. SH-SY5Y cells were classified into normal control group， OGD/R group， and low-， medium-， and high-dose SP （2∶1） extract groups （10， 30， 100 mg·L-1， respectively）. Cells in the groups， except the normal control group， were rapidly reoxygenated for 12 h after 4 h OGD for modeling. Then cell viability was detected by CCK-8 and cell morphology was observed under the microscope. The release rate of lactate dehydrogenase （LDH）， superoxide dismutase （SOD） activity， and content of glutathione （GSH） and malondialdehyde （MDA） were determined by spectrophotometry. The level of reactive oxygen species （ROS） was detected with 2，7-dichlorodihydrofluorescein diacetate （DCFH-DA） and mitochondrial membrane potential with JC-1 assay. The nuclear morphology was observed based on Hoechst 33342 staining， and apoptosis was examined by flow cytometry combined with Annexin V-FITC/PI staining.ResultThe viability of the cells was highest in the presence of the extract of the two medicinals mixed at the ratio of 2∶1. Compared with normal control group， OGD/R group showed damaged cell morphology， high release rate of LDH and levels of ROS and MDA （P<0.01）， low SOD activity and GSH level （P<0.01）， low mitochondrial membrane potential， and high apoptosis rate （P<0.01）. Compared with OGD/R group， SP extract improved cell viability and cell morphology and reduce cell LDH release rate in a concentration-dependent manner （P<0.01）. In addition， SP extract at 30， 100 mg·L-1 reduced the level of intracellular ROS and increased SOD activity and GSH level （P<0.05， P<0.01）， and SP extract at 100 mg·L-1 decreased the content of MDA （P <0.05）. Moreover， SP extract increased mitochondrial membrane potential， and SP extract at 30， 100 mg·L-1 lowered the apoptosis rate （P<0.01）.ConclusionThe extract of Salvia miltiorrhiza Bunge and Radix Puerariae mixed at 2∶1 shows better protective effect on OGD/R-injured SH-SY5Y cells. The mechanism is the likelihood that it alleviates oxidative damage of cells and inhibits cell apoptosis.
Keywords：ischemic stroke;oxygen-glucose deprivation/reoxygenation （OGD/R） model;Salviae Miltiorrhizae Radix et Rhizoma and Puerariae Lobatae Radix;oxidative stress;apoptosis
Abstract：ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo.MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry.ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency.ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
Abstract：ObjectiveTo explore the pharmacodynamic effect of the water extract of Citri Grandis exocarpium （WEC） on mice with alcohol-induced acute liver injury and provide data support for the development of this medicinal for anti-alcoholism and liver protection.MethodThe main components of WEC were determined by high performance liquid chromatography （HPLC）. Sixty Balb/c mice were randomized into 6 groups： control group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， model group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， low-， medium-， and high-dose WEC groups （0.5， 1.0， 2.0 g·kg-1）， and Haiwang Jinzun tablet positive control group （2.0 g·kg-1）. The administration lasted 14 days. One day before the end of the administration， mice were fasted for 12 h with free access to water. The mice， except the control group， were given 56° Chinese liquor （13 mL·kg-1）. After 2 h， blood was taken from eyeballs and the liver was dissected and weighed. Automatic biochemical analyzer was employed to detect the expression of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and alcohol dehydrogenase （ADH）. The pathological changes of liver tissues were observed based on hematoxylin-eosin （HE） staining， and apoptosis of hepatocytes based on TUNEL/DAB staining. The expression of proteins related to apoptosis was detected by Western blot.ResultAccording to the HPLC fingerprint， the main components of WEC were rhoifolin and naringin. Compared with the control group， the model group showed increase in liver/body weight ratio （P<0.01） and the expression of ALT and AST （P<0.05， P<0.01）， decrease in the expression of ADH （P<0.05）， blurred structure of hepatic lobules， pathological changes of liver tissue， loose cytoplasm with edema， severe steatosis， rise of the TUNEL-positive rate （P<0.01）， reduction in expression of Bcl-2 （P<0.01）， and increase in Bax and Caspase-3 （P<0.01）. Compared with the model group， medium-dose WEC lowered liver/body weight ratio （P<0.05）. All doses of WEC depressed the activity of ALT and AST （P<0.05， P<0.01）， up-regulated the expression of ADH （P<0.05）， significantly improved the pathological features of alcohol-induced cytoplasmic porosity， edema， and steatosis， down-regulated the TUNEL-positive rate （P<0.05， P<0.01）， enhanced the expression of Bcl-2 （P<0.05）， and decreased Bax and Caspase-3 （P<0.01）.ConclusionWEC regulates the expression of ALT， AST， and ADH and improves hepatic steatosis and hepatocyte apoptosis to fight against acute liver injury.
Abstract：ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide （LPS） injection， ligation of coronary artery and different combinations of the two methods.MethodA total of 225 male SD rats were randomly divided into sham operation group， simple coronary artery ligation group， first injected LPS group ［LPS（5 mg·kg）injection 24 h before coronary artery ligation］ and follow injected LPS group ［LPS（5 mg·kg）injection 10 min after coronary artery ligation］. The indexes of each group were detected at 3， 24， 72 h after modeling， and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine （TCM） syndrome （tongue and pulse） of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function， and the myocardial ischemia and infarction areas were measured by Evans blue/2，3，5-triphenyltetrazolium chloride （TTC） staining. The content of creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， creatine kinase （CK）， and troponin T （cTnT） in serum as well as interleukin-1 β （IL-1β） and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay （ELISA）. Hematology analyzer was adopted to determine the white blood cell （WBC） count， and the four coagulation indexes， platelet aggregation rate， hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin（HE）staining.ResultAfter 3 h of modeling， compared with the conditions in sham operation group， the R， G and B values of tongue of rats （P<0.01）， pulse amplitude （P<0.01）， and cardiac function in simple coronary artery ligation group were decreased， and the color of hypoglossal veins became purple（P<0.01）. The content of CK， LDH， cTnT， IL-1β and IL-6 in serum（P<0.05）， myocardial infarction area（P<0.01）， and total number of WBCs （P<0.05）were increased. Compared with simple coronary artery ligation group， first injected LPS group and follow injected LPS group had increased hypoglossal veins， decreased R value of tongue and elevated cTnT content （P<0.01）， while follow injected LPS group had reduced B value of tongue， decreased cardiac output （CO）（P<0.05）， increased IL-1β content， and thinned left ventricular anterior walls at end-systole （LVAWs）（P<0.01）. After 24 h of modeling， compared with sham operation group， simple coronary artery ligation group presented significantly decreased R， G and B values of tongue， lengthened purplish dark hypoglossal veins （P<0.01）， reduced pulse amplitude（P<0.01） and cardiac function， enlarged myocardial infarction area（P<0.01）， increased whole blood viscosity， platelet aggregation rate， fibrinogen （FIB）， shortened prothrombin time （PT） and thrombin time （TT）（P<0.01）， and elevated total number of WBCs （P<0.01）and content of CK， LDH， cTnT and IL-6 in serum（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the pulse amplitude， R， G and B values of tongue （P<0.01）， and ejection fraction （EF） and fractional shortening （FS） scores （P<0.05）dropped， and hypoglossal veins were deepened and lengthened（P<0.05）， and cTnT content was increased（P<0.01）in first injected LPS group and follow injected LPS group. However， follow injected LPS group had thinned LVPWs， increased LDH content， platelet aggregation rate（P<0.05）， myocardial infarction area， and total number of WBC， level of IL-1β（P<0.05）， and shortened TT（P<0.01）. Additionally， 72 h after modeling， compared with sham operation group， simple coronary artery ligation group showed significantly reduced pulse amplitude， lowered R， G and B values of tongue， thickened and lengthened hypoglossal veins（P<0.01）， decreased cardiac function， and increased content of cTnT， FIB， whole blood viscosity（P<0.01），platelet aggregation rate， level of IL-6 and IL-1β（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple， and the content of cTnT was boosted（P<0.01）， whereas follow injected LPS group had decreased pulse amplitude， R， G and B values of tongue， EF and FS scores （P<0.05）， and enlarged myocardial infarction area（P<0.01）.ConclusionCompared with the other modeling methods and models at different modeling time， the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.
Keywords：animal model;acute coronary syndrome;combined blood stasis and poison;disease-syndrome combination
Abstract：ObjectiveTo investigate the possible mechanism of different doses of L-Borneolum，Borneolum，and Borneolum Syntheticum in the electrophysiology，anti-inflammation，and regulation of hypoxia inducible factor-1α （HIF-1α）/vascular endothelial growth factor （VEGF） cardiovascular protection of the experimental acute myocardial infarction （AMI） rats.MethodSD male adult rats were randomly divided into thirteen groups according to their body weight，namely the sham operation group，the model group，the solvent model group，the nitroglycerin group，the Borneolum high，medium，and low-dose （0.6，0.3， 0.15 g·kg-1） groups，the L-Borneolum high，medium，and low-dose （0.2，0.1， 0.05 g·kg-1） groups，and the Borneolum Syntheticum high，medium，and low-dose （0.2，0.1， 0.05 g·kg-1） groups，with 10 rats in each group. Rats were given 10 mL·kg-1 by gavage for 3 d of pre-administration. Thirty minutes after the last administration，the left anterior descending coronary artery （LAD） was ligated to induce the model，and the successful rat model was continuously treated for 3 d. BL-420N biosystem was used to analyze the electrocardiogram （ECG） and heart rate variability （HRV） before and after modeling and after 3 d of treatment. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to determine the mRNA expressions of interleukin-1β （IL-1β） and interleukin-6 （IL-6） in the myocardial tissue Western blot and immunohistochemistry were used to determine the protein expression levels of VEGF receptor 1 （VEGFR1），HIF-1α，and CD34.ResultCompared with the sham operation group，the model group significantly increased the heart rate，ECG ST wave，T wave，QRS duration，QTC interval，and Q wave on the day of modeling and after 3 d of treatment，and significantly changed HRV and T wave （P<0.05，P<0.01）. As compared with the solvent model group，on the day of modeling，the heart rate of the L-Borneolum medium and low-dose groups and the Borneolum groups，the ST wave of the L-Borneolum groups，the Borneolum high and medium-dose groups，and the Borneolum Syntheticum high-dose group，HRV parameters of the L-Borneolum groups，the Borneolum medium and low-dose groups，and the Borneolum Syntheticum high-dose group，LF/HF of the L-Borneolum high and medium-dose group，the Borneolum low-dose group，and the Borneolum Syntheticum groups，T wave of the L-Borneolum high-dose group，the Borneolum Syntheticum high-dose group，and Borneolum medium-dose group，QTC interval of the L-Borneolum medium and low-dose groups and the Borneolum high and medium-dose groups，and QRS duration of the L-Borneolum high and low-dose groups，the Borneolum high and low-dose groups，and the Borneolum Syntheticum groups were significantly reduced or shortened （P<0.05，P<0.01）. After 3 d of treatment，the heart rate of the L-Borneolum groups，the Borneolum high and medium-dose groups，and the Borneolum Syntheticum medium-dose group，ST wave of the L-Borneolum group，the Borneolum high and medium-dose groups，and the Borneolum Syntheticum high-dose group，OTC interval，ORS duration，and Q wave of the L-Borneolum high-dose group，the Borneolum high-dose group，and the Borneolum Syntheticum high and medium-dose groups，QRS duration of the L-Borneolum medium-dose group，QTC interval of the Borneolum medium-dose group，and Q wave of the Borneolum Syntheticum low-dose group were all significantly reduced or shortened（P<0.01）. The mRNA expressions of IL-1β and IL-6 in the L-Borneolum medium and low-dose groups，the Borneolum medium and low-dose groups，and the Borneolum Syntheticum high and medium-dose groups were significantly down-regulated（P<0.01），and LF/HF in the L-Borneolum high and medium-dose groups，the Borneolum high and medium-dose groups，and the Borneolum Syntheticum high and low-dose groups were significantly reduced （P<0.05，P<0.01）. HRV in the L-Borneolum high-dose group，the Borneolum groups，and the Borneolum Syntheticum high and low-dose groups，and T wave in the Borneolum high and medium-dose groups and the Borneolum Syntheticum high-dose group were increased significantly. The protein expressions of HIF-1α，VEGFR1，and CD34 in the L-Borneolum medium and low-dose groups，the Borneolum low-dose group，and the Borneolum Syntheticum high-dose group were significantly up-regulated，as well as those of VEGFR1 and CD34 in the Borneolum medium-dose group （P<0.05，P<0.01）.ConclusionThe 3 kinds of Borneolum improves the heart rate，heart rate variability，and electrocardiogram of AMI model rats to different degrees，and may play a myocardial protective effect by anti-inflammation and promotion of angiogenesis. The combined effect suggests that L-Borneolum has the superior effect next to Borneolum，and Borneolum Syntheticum has the inferior effect.
Keywords：3 kinds of Borneolum;myocardial infarction;myocardial electrophysiology;inflammatory factors;angiogenesis
Abstract：ObjectiveTo screen out the extended spectrum beta-lactamase （ESBL）-producing Escherichia coli with the strongest biofilm-forming ability through experiments， and discuss the effect of modified Dayuansan （MDYS） combined with imipenem-cilastatin and cilastatin sodium on the biofilm of E. coli.MethodThe paper diffusion and crystal violet staining methods were used to identify 19 clinically isolated strains of drug-resistant E. coli-induced enzymes and the biofilm-forming ability. The induced enzymes and the E. coli with the strongest biofilm-forming ability were screened out. The minimum inhibitory concentration （MIC） value of MDYS and imipenem-cilastatin and cilastatin sodium was determined by 2，3-bis（2-methoxy-4-nitro-5-sulfophenyl）-2H-tetrazolium-5-carboxamide （XTT） assay. The 1/2， 1/4， and 1/8 MIC of the water extract of MDYS， imipenem-cilastatin and cilastatin sodium alone， and MDYS combined with imipenem-cilastatin and cilastatin sodium was determined by methyl thiazolyl tetrazolium （MTT） assay to obtain the optimum concentration of drugs. BioFlux dynamically observed the effect of the optimum combined drug concentration on the number of bacteria in the biofilm and the biofilm formation of E. coli， and observed the distribution of live/dead bacteria with a laser confocal scanning microscope. Finally， the morphological changes in bacteria after drug treatment were observed statically by scanning electron microscopy.ResultE5E7 strain was ESBL enzyme and the E. coli with the strongest biofilm-forming ability. The results of MTT assay showed that the MIC values of the water extracts of imipenem-cilastatin and cilastatin sodium and MDYS were 1 mg·L-1 and 250 g·L-1， respectively. The results of XTT assay showed that compared with the blank group， the 1/2， 1/4， and 1/8 MIC MDYS groups and the combined drug groups significantly decreased the number of bacteria in the biofilm （P<0.01）. The inhibitory effect diminished as the concentration of imipenem-cilastatin and cilastatin sodium decreased. Compared with the imipenem-cilastatin and cilastatin sodium group with the same concentration， the combined drug group improved the inhibitory effect on the number of bacteria in the biofilm （P<0.01）. Compared with the MDYS group with the same concentration， 1/2 MIC imipenem-cilastatin and cilastatin sodium combined with 1/2， 1/4， and 1/8 MIC MDYS， 1/4 MIC imipenem-cilastatin and cilastatin sodium combined with 1/2 and 1/4 MIC MDYS， and 1/8 MIC imipenem-cilastatin and cilastatin sodium combined with 1/2 and 1/4 MIC MDYS decreased the number of bacteria （P<0.05， P<0.01）. The results of BioFlux showed that compared with the blank group， the 1/2 and 1/8 MIC imipenem-cilastatin and cilastatin sodium groups had an insignificant effect on the area of biofilm， whereas the 1/2 and 1/4 MIC MDYS groups significantly decreased the area of biofilm. The results under the scanning electron microscopy showed that as compared with the blank group and the imipenem-cilastatin and cilastatin sodium group， the division cycle was significantly longer under the action of MDYS combined with imipenem-cilastatin and cilastatin sodium. The length of the division cycle in the combined drug group was higher than that in drug alone group.ConclusionIn vitro studies reveal that MDYS combined with commonly-used antibiotics can inhibit the biofilm status of multi-drug resistant E. coli， and MDYS has the effect of enhancing sensitization and inhibiting bacteria with synergistic antibiotics.
Abstract：ObjectiveTo explore the effect of Scutellariae Barbatae Herba extract on on the cycle arrest of nasopharyngeal carcinoma cells and the possible mechanism by adding different concentration of Scutellariae Barbatae Herba extract （0.25， 0.5， 1 g·L-1） in the culture medium， taking CNE1 （nasopharyngeal carcinoma cells） as the research object.MethodAfter the treatment of CNE1 by Scutellariae Barbatae Herba extract， cell counting kit-8 （CCK-8） was used to detect cell proliferation， and Giemsa staining was used to detect the clone formation rate. Flow cytometry was used to detect cell cycle distribution， and reverse transcription-polymerase chain reaction （ RT-PCR） assay and Western blot assay were used to detect the relative expression of messenger ribonucleic acid （mRNA） by small interfering RNA （siRNA） or overexpression.ResultAs compared with the blank group， the proliferation and colony formation rate of CNE1 in the Scutellariae Barbatae Herba extract group significantly decreased （P<0.05， P<0.01） in a dose and time-dependent manner， whereas the percentage of cells in the presynthetic phase （G0/G1） increased （P<0.05， P<0.01）. The expression level of S-phase kinase associated protein 2 （SKP2） in the Scutellariae Barbatae Herba extract group significantly decreased （P<0.01） as compared with the blank group. As compared with the Scutellariae Barbatae Herba extract group， the protein levels of p21 and p27 significantly decreased in the overexpressed SKP2+ Scutellariae Barbatae Herba extract group （P<0.01）. As compared with the blank group， the signal activation and the phosphorylation level of signal transducer and activator of transcription 3 （STAT3） of CNE1 in the S. barbata extract group significantly decreased （P<0.05， P<0.01）.ConclusionScutellariae Barbatae Herba extract effectively inhibits the proliferation of CNE1， and the mechanism may be related to its action on the STAT3/SKP2 pathway.
Keywords：Scutellariae Barbatae Herba;nasopharyngeal carcinoma;cell cycle arrest;signal transducer and activator of transcription 3 （STAT3）;S-phase kinase associated protein 2 （SKP2）
Abstract：ObjectiveTo investigate the effect of Shengjiang Tonglong prescription hollow suppository on rats with prostate hyperplasia， and the effect of the proteins related to phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） signaling pathway in the prostate， thus exploring the mechanism of Shengjiang Tonglong prescription hollow suppository in the treatment of rats with prostate hyperplasia.MethodTen SD male rats were randomly selected from 60 SD male rats to form a sham operation control group， and the rest rats were subcutaneously injected with testosterone propionate for 4 consecutive weeks after castration to induce the rat model of prostatic hyperplasia. According to the random number table method， the 50 rats were randomly divided into a model group， a finasteride group （0.45 mg·kg-1）， and three high， middle， and low-dose Shengjiang Tonglong prescription hollow suppository groups （3.98， 1.99， 0.99 g·kg-1）， with ten rats in each group. After castration for 7 d， the sham operation control group and the model group used the blank hollow suppositories， and the finasteride group and the Shengjiang Tonglong prescription hollow suppository groups used the corresponding hollow suppositories. The drugs were given to the rats by anal plugs continuously for 28 d. The rats were then killed， and the prostate tissues were separated and weighed to observe the effects of drugs on the prostate index of rats in each group. The hematoxylin-eosin （HE） staining was used for the pathological observation of the prostate tissues. The level of dihydrotestosterone （DHT） was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of PI3K/Akt signaling pathway protein， B-cell lymphoma-2 （Bcl-2）， cysteine aspartate-specific protease-3 （Caspase-3）， Bcl-2-associated X protein （Bax）， and αB-crystallin （CRYAB） protein in the prostate tissues.ResultAs compared with the sham operation control group， the protein expression levels of prostate index， DHT level， CRYAB， Bcl-2， PI3K， and Akt in the model group were increased， and the protein expression levels of Caspase-3 and Bax were decreased （P<0.05， P<0.01）. As compared with the model group， the prostate index in the high-dose Shengjiang Tonglong prescription hollow suppository group was decreased （P<0.05）， and the level of DHT and the protein expression levels of CRYAB， Bcl-2， PI3K， and Akt in the prostate of the Shengjiang Tonglong prescription hollow suppository groups were decreased， and the protein expression levels of Caspase-3 and Bax were increased （P<0.05， P<0.01）.ConclusionShengjiang Tonglong prescription hollow suppository decreases the expression of CRYAB protein， negatively regulates the PI3K/Akt signaling pathway， down-regulates the level of DHT and the protein expression levels of Bcl-2， PI3K， and Akt， and up-regulates the protein expression levels of Caspase-3 and Bax， thereby inhibiting cell proliferation and promoting cell apoptosis， which plays a therapeutic role in the benign prostate hyperplasia （BPH）. The high-dose Shengjiang Tonglong prescription hollow suppository significantly improves prostatic hyperplasia in rats.
Abstract：ObjectiveTo study the effect of apigenin on the proliferation and apoptosis of human colon cancer CL187 cells and the underlying mechanisms.MethodHuman colorectal cancer CL187 cells were treated with different concentrations of apigenin （0， 30， 45， 60 mg·L-1） according to the results of the preliminary experiment. The proliferation of CL187 cells was detected by methyl thiazolyl tetrazolium （MTT） and colony formation assays， and the apoptosis was observed via Hoechst 33258 staining. Real-time fluorescence quantitative PCR was conducted to determine the mRNA levels of cysteine protease-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the CL187 cells treated with apigenin. Western blot was employed to measure the protein levels of Caspase-3， Bcl-2， and Bax associated with apoptosis， protein kinase B （Akt） and phosphorylated Akt （p-Akt） in phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathway， and extracellular signal-regulated kinases 1/2 （ERK1/2）， p-ERK1/2， c-Jun N-terminal kinase （JNK）， p-JNK， p38 mitogen-activated protein kinase （MAPK）， and p-p38 MAPK protein in MAPK pathway.ResultCompared with the blank group， the apigenin groups had low cell survival rates and high inhibition rates on cell proliferation （P<0.01）. Apigenin decreased the cell clone number and clone formation rate， and increased the inhibition rate on clone formation （P<0.01）. After CL187 cells were treated with different concentrations of apigenin for 48 h， typical apoptosis characteristics such as nuclear pyknosis， chromatin condensation， and enhanced fluorescence reaction were observed. Compared with blank group， 45， 60 mg·L-1 apigenin treatments down-regulated the mRNA level of anti-apoptotic gene Bcl-2 （P<0.01） and all the apigenin treatments up-regulated those of the pro-apoptotic genes Bax and Caspase-3 （P<0.05， P<0.01）. Similarly， apigenin treatments down-regulated the protein level of Bcl-2 （P<0.05， P<0.01） and up-regulated those of Caspase-3 （P<0.05， P<0.01） and Bax （P<0.01， 45， 60 mg·L-1）. The blank group had higher protein level of Akt than the 60 mg·L-1 apigenin group （P<0.01）， higher protein levels of p-Akt， ERK1/2， and p-ERK1/2 than the 45， 60 mg·L-1 apigenin groups （P<0.01）， and higher protein levels of JNK and p-JNK than the apigenin groups （P<0.05， P<0.01）. Compared with blank group， 60 mg·L-1 apigenin up-regulated the protein level of p38 MAPK （P<0.05）， and all the apigenin groups up-regulated that of p-p38 MAPK （P<0.01）. Furthermore， apigenin lowered the p-Akt/Akt ratio （P<0.05， P<0.01） and p-ERK1/2/ERK1/2 ratio （P<0.01）， while it increased the p-JNK/JNK ratio （45， 60 mg·L-1； P<0.05， P<0.01） and p-p38 MAPK/p38 MAPK ratio （P<0.05， P<0.01）.ConclusionApigenin can inhibit the proliferation and promote the apoptosis of CL187 cells by inhibiting the PI3K/Akt signaling pathway and regulating the expression of proteins in the MAPK signaling pathway.
Keywords：apigenin;colon cancer;CL187 cells;proliferation;apoptosis;phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway;mitogen-activated protein kinase （MAPK） signaling pathway
Abstract：ObjectiveTo evaluate the efficacy and safety of An'erning granules in the treatment of community-acquired pneumonia in children.MethodA randomized， double-blind， single-simulation， placebo-controlled trial was designed in this study. The children were randomly assigned into an observation group （An'erning granules combined with ceftriaxone sodium） and a control group （An'erning granules placebo combined with ceftriaxone sodium） according to the ratio of 2∶1. The disease cure rate was taken as the main indicator of efficacy， and the safety of An'erning granules was observed.ResultA total of 206 children （137 in the observation group and 69 in the control group） were included in this study. Before treatment， the age， sex， body height， body weight， diagnosis time of pneumonia， and symptom and sign scores had no significant differences between the two groups. After 8 days of continuous medication， the observation group［70.80%（97/137）］ had higher cure rate than the control group［56.52%（39/69）］（χ2=4.17，P<0.05） and total effective rate of chest X-ray ［97.98%（97/99）］ than the control group［86.27%（44/51）］ （χ2=12.98，P<0.01）. The observation group was superior to the control group in the alleviation of TCM syndrome under the condition of 0-3 g dose stratification on day 3 of medication （P<0.01）. The recovery time， time to complete fever abatement， time to fever abatement and expectoration alleviation， rate of conversion to severe case， and reduction in the frequency of antibiotic use showed no significant differences between the two groups. In terms of safety， 13 and 7 adverse events occurred in the observation group and control group， respectively， which were relieved or disappeared after drug withdrawal or symptomatic treatment and showed no significant difference between the two groups.ConclusionIntravenous drip of ceftriaxone sodium combined with An'erning granules is effective in the treatment of community-acquired pneumonia in children. It can accelerate the absorption of pulmonary inflammation， alleviate the clinical symptoms in a short time for young children or the children with mild symptoms， and is safe in clinical application.
Keywords：An'erning granules;community-acquired pneumonia in children;randomized controlled trial;antibiotic
Abstract：ObjectiveTo explore the effect of Suanzaoren Tang combined with Ziwu Liuzhu acupuncture on the vertebral artery hemodynamics， inflammatory cytokines， and neurotrophic factors in the patients with cervical insomnia with syndrome of deficiency of both heart and spleen.MethodThe random number table method was employed to assign 164 patients with cervical insomnia with syndrome of deficiency of both heart and spleen treated in the First Clinical Medical School of Guangzhou University of Chinese Medicine from January 2018 to June 2021 into a control group and an observation group. The control group was orally administrated with 1-2 mg estazolam tablets before bed for 4 weeks， and the observation group with Suanzaoren Tang combined with Ziwu Liuzhu acupuncture for 4 weeks. The therapeutic efficacy and safety were observed. The Pittsburgh Sleep Quality Index （PSQI） score， polysomnography monitoring results， hemodynamics parameters of vertebral artery， and serum levels of inflammatory cytokines and neurotrophic factors were compared before and after treatment.ResultExcept 4 dropouts， the remaining 160 patients were included in this study， with 80 patients in each group. The observation group had higher total effective rate than the control group ［92.50% （74/80） vs. 80.00% （64/80）， χ2=5.270， P<0.05］. Compared with that before treatment， the therapies in both groups decreased the PSQI score， sleep latency time， awakening time， awakening times， serum levels of interleukin-1β （IL-1β）， C-reactive protein （CRP）， and tumor necrosis factor-α （TNF-α） （P<0.01）. Meanwhile， they increased the proportion of rapid-eye-movement （REM） sleep， the diastolic blood flow velocity （Vd）， systolic blood flow velocity （Vs）， and mean blood flow velocity （MFV） of vertebral artery， as well as the serum levels of brain-derived neurotrophic factor （BDNF） and glial cell-derived neurotrophic factor （GDNF） （P<0.05， P<0.01）. Moreover， the observation group had lower PSQI score， sleep latency time， awakening time， awakening times， and serum IL-1β， CRP， and TNF-α levels （P<0.01） and higher proportion of REM sleep， Vd， Vs， MFV of vertebral artery， and serum BDNF and GDNF levels （P<0.05， P<0.01） than the control group.ConclusionZiwu Liuzhu acupuncture combined with Suanzaoren Tang can improve blood circulation of vertebral artery， reduce the serum levels of inflammatory cytokine， and increase the serum levels of neurotrophic factors to improve the sleep quality of the patients with cervical insomnia with syndrome of deficiency of both heart and spleen.
Abstract：ObjectiveTo evaluate the success of Qi-deficiency model of Balb/C-nu mice established by swimming exhaustion test from the view of biomarkers and metabolic pathways by metabonomics.MethodBalb/C-nu mice were randomly divided into the normal group and Qi-deficiency group， Qi-deficiency model was established by swimming with 5% body weight metal fixed at the tail for 15 consecutive days until exhaustion （nose tip immersion time>5 s）. The urine metabonomics was analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MS）， and the mobile phase was acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-1 min， 5%-8%A； 1-4 min， 8%-8.5%A； 4-5 min， 8.5%-12%A； 5-10 min， 12%-40%A； 10-12 min， 40%-100%A； 12-15 min， 100%A）， the flow rate was 0.3 mL·min-1， the injection volume was 10 μL， electrospray ionization （ESI） in positive and negative ion modes was used in MS analysis， the MS data were acquired in full-scan mode from m/z 50-1 000. Principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， human metabolome database （HMDB）， high collision energy ion fragments collected by MSE and tandem MS （MS/MS） ion information were used to identify potential biomarkers. Kyoto Encyclopedia of Genes and Genomes （KEGG） database and MetaboAnalyst 5.0 were used to analyze the corresponding metabolic pathways and pathway enrichment of biomarkers.ResultEndogenous substances in urine of mice in normal group and Qi-deficiency group were obviously separated， and there were 24 biomarkers with significant difference. The metabolic pathways involved in these biomarkers were tricarboxylic acid cycle， glycolysis metabolism， amino acid metabolism， fatty acid metabolism， pyrimidine metabolism， steroid hormone biosynthesis and tryptophan metabolism. Among them， the metabolic pathways related to energy were tricarboxylic acid cycle， glycolysis metabolism， amino acid metabolism， fatty acid metabolism， pyrimidine metabolism and steroid hormone biosynthesis.ConclusionThrough the investigation of urine metabonomics， combined with the physical signs， the Qi-deficiency model established by swimming exhaustion test in Balb/C-nu mice is successfully evaluated， and it is also verified that there is a close correlation between Qi-deficiency and energy metabolism.
Keywords：Qi-deficiency;metabonomics;energy metabolism;ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MS）;traditional Chinese medicine syndrome;Balb/C-nu mice;mechanism
Abstract：ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine.MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4.ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4.ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.
Keywords：Morindae Officinalis Radix;kidney Yang deficiency model;hypothalamic-pituitary-gonadal axis （HPG axis）;E-cadherin;α-smooth muscle actin;Wnt2b;β-catenin;Glycyrrhizae Radix et Rhizoma
Abstract：ObjectiveTo explore the correlation between dryness and energy metabolism of Atractylodis Rhizoma， and to analyze the difference of medicinal properties between Atractylodes lancea and A. chinensis.MethodA total of 110 healthy SD rats were randomly divided into 11 groups， including normal group， volatile oil of A. lancea 1-5 group （S1-S5 group， doses of 447， 473， 442， 489， 496 mg·kg-1） and volatile oil of A. chinensis 1-5 group （N1-N5 group， doses of 197， 118， 281， 222， 185 mg·kg-1）， the administration volume was 0.01 mL·g-1 with intragastric administration for 21 days. Dryness effect of A. lancea and A. chinensis on rats was evaluated by comparing the body weight， drinking water volume， urine volume， whole blood viscosity and pathological sections of submandibular gland stained with hematoxylin-eosin （HE）. The expression of aquaporin 2 （AQP2） in rat kidney was measured by immunohistochemistry， the mRNA expressions of cytochrome C oxidase subunit 7A2 （COX7A2） and succinate dehydrogenase （SDH） complex subunit D （SDHD） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The contents of SDH， lactate dehydrogenase （LDH） and sodium ion-potassium ion-adenosine triphosphatase （Na+-K+-ATPase） in rat plasma were determined by colorimetry. The quality of A. lancea and A. chinensis was evaluated by coefficient of variation method， and Pearson correlation coefficient was used to analyze the correlation between dryness and energy metabolism.ResultCompared with the normal group， the amounts of drinking water and urine in volatile oil of A. lancea group and volatile oil of A. chinensis group increased， and the submandibular gland acini atrophied， the whole blood viscosity of rats in the volatile oil of A. lancea group increased significantly （P<0.01）， the expression levels of COX7A2 and SDHD mRNA， the activities of SDH， LDH and Na+-K+-ATPase increased significantly （P<0.01）， and the expression of AQP2 in kidney decreased significantly （P<0.01）. Compared with the normal group， the expression level of COX7A2 mRNA， SDH activity and whole blood viscosity in the volatile oil of A. chinensis group increased （P<0.05， P<0.01）， the AQP2 and SDH mRNA expression levels， LDH and Na+-K+-ATPase activities had no significant difference. The comprehensive score analysis of each index showed that the effect of volatile oil of A. lancea on dryness and energy metabolism was stronger than that of volatile oil of A. chinensis， and there was a positive correlation between dryness index and energy metabolism index.ConclusionThe two indexes show that medicinal properties of A. lancea is stronger than that of A. chinensis， and energy metabolism is closely related to the dryness of Atractylodis Rhizoma. It is suggested that it is reasonable to evaluate the dryness effect of Atractylodis Rhizoma from the perspective of energy metabolism， which can further enrich the evaluation indexes of medicinal properties.
Abstract：ObjectiveTo compare the differences in resistance and structure of skin between acupoints and non-acupoints， and to study the difference in skin permeability characteristics of Corydalis Rhizoma total alkaloid patches （CTTP） after administration at Shenque acupoint and non-acupoint， so as to provide experimental support for its clinical acupoint application to prevent and treat chronic pain.MethodTaking corydaline （CD）， tetrahydropalmatine （THP） and corydalis L （CDL） as evaluation indexes， and the quantitative analysis was carried out by high performance liquid chromatography （HPLC）. The mobile phase was methanol-0.04 mol·L-1 phosphoric acid aqueous solution （70∶30， pH 6.0 adjusted with triethylamine）， the detection wavelength was 281 nm. In vitro transdermal test in Franz diffusion cell and in vivo transdermal test were used to study the skin permeability characteristics of CTTP through Shenque acupoint and non-acupoint administration. At the same time， the skin resistance between Shenque acupoint and non-acupoint was measured before and after the administration， and the distribution of the drug in each layer of the skin was compared by freezing sectioning， and visual verification was performed with fluorescence inverted microscope.ResultAfter 24 h of administration， the results of in vivo and in vitro experiments showed that the cumulative permeation and retention of CD， THP and CDL at Shenque acupoint skin were higher than those at non-acupoint skin （P<0.05， P<0.01）， the skin resistance of Shenque acupoint was lower than that of non-acupoint at all time points. The fluorescence microscopic observation results showed that the drug content of each layer of the skin was all Shenque acupoint>non-acupoint， indicating that the skin of Shenque acupoint had better effect on drug penetration and storage than non-acupoint.ConclusionThe 24 h cumulative permeation and retention of CTTP in Shenque acupoint skin are higher than those in non-acupoint skin， and the mechanism may be related to the thin skin， low electrical resistance and large number of hair follicle bodies at Shenque acupoint.
Abstract：ObjectiveTo identify the chemical constituents of Alismatis Rhizoma before and after processing with salt-water by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）， and to investigate the changes of terpenoids in Alismatis Rhizoma before and after processing with salt-water.MethodUPLC-Q-TOF-MS was used to detect with 0.1% formic acid aqueous solution （A）-acetonitrile （B）as mobile phase for gradient elution （0-0.01 min， 20%B； 0.01-5 min， 20%-40%B； 5-40 min， 40%-95%B； 40-42 min， 95%B； 42-42.1 min， 95%-20%B； 42.1-45 min， 20%B）， electrospray ionization （ESI） was selected for collection and detection in positive ion mode with the scanning range of m/z 100-1 250 and ion source temperature at 500 ℃. The data were analyzed by PeakView 18.104.22.168， the components were identified according to the primary and secondary MS data， and combined with the reference substance and literature. After normalized treatment by MarkerView 1.2.1， the MS data were analyzed by principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）， and then the differential components before and after processing were screened. The content changes of differential components were analyzed according to the relative peak area.ResultA total of 30 components were identified under positive ion mode， including 28 prototerpene triterpenes and 2 sesquiterpenes. The results of PCA and OPLS-DA showed that there were significant differences in components from Alismatis Rhizoma before and after processing with salt-water， and 10 differential components （alisol B 23-acetate， alisol I， alismol， 11-deoxy-alisol B 23-acetate， alisol B， alisol C， 11-deoxy-alisol B， alisol G， 11-deoxy-alisol C and alisol A） were screened， and the contents of alisol G and alisol A decreased significantly after processing.ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents in raw and salt-processed products of Alismatis Rhizoma. It takes a great difference in the contents of chemical constituents before and after processing， and the difference of substituents is the main reason for this differences， which can provide reference for determining the material basis of efficacy changes of Alismatis Rhizoma before and after processing with salt-water.
Keywords：processing of traditional Chinese medicine;Alismatis Rhizoma;ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）;salt-processed products;terpenoids;principal component analysis （PCA）;orthogonal partial least squares-discriminant analysis （OPLS-DA）
Abstract：ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp.MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively.ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution.ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.
Keywords：hemp;type 2C protein phosphatases （PP2C）gene family;gene identification;functional prediction;expression analysis
Abstract：Shengyang Yiweitang is one of the first 100 classical prescriptions published by the National Administration of Traditional Chinese Medicine. It originated from the Clarifying Doubts about Damage from Internal and External Causes by physician LI Dongyuan of Jin dynasty， and is composed of Astragali Radix， Ginseng Radix et Rhizoma， Glycyrrhizae Radix et Rhizoma， Atractylodis Macrocephalae Rhizoma， Poria， Pinelliae Rhizoma， Citri Reticulatae Pericarpium， Angelicae Pubescentis Radix， Saposhnikoviae Radix， Notopterygii Rhizoma et Radix， Bupleuri Radix， Paeoniae Radix Alba， Alismatis Rhizoma， and Coptidis Rhizoma. With the effects of replenishing Qi， promoting Yang， clearing heat and removing dampness， Shengyang Yiweitang is used to treat spleen-stomach weakness and dampness-heat accumulation syndrome. Using bibliometrics， the authors systematically sorted out the source，composition， dosage， preparation， efficacy， indications， principle of composition， origin and processing of drugs，and modern clinical application of the prescription， and explored its history and key information. Additionally， it was found that Shengyang Yiweitang was widely used in modern clinical practice and was suitable for multisystem diseases， of which digestive system （264） was the most common， accounting for 41.71%， followed by urogenital system （57， 9.00%） and nervous system （48， 7.58%）. Although the treatment scope was wide， the pathogenesis of the diseases in traditional Chinese medicine belongs to "spleen-stomach weakness"， which fully reflected Li's academic thought of "internal injury of spleen and stomach leads to various diseases". The key information of Shengyang Yiweitang was determined by summarizing the relevant ancient books and modern literature， so as to provide accurate reference for its rational clinical application and further research and development.
Keywords：famous classical prescription;Shengyang Yiweitang;historical evolution;clinical application;key information;literature research
Abstract：ObjectiveTo predict the potential molecular mechanism of Erxian decoction in the treatment of anxiety disorder based on network pharmacology， and to verify the efficacy and mechanism using the animal model of maternal separation combined with restraint stress.MethodActive components and related targets of Erxian decoction were obtained by traditional Chinese medicine system pharmacology database and analysis platform （TCMSP） and SwissTargetPrediction. The targets related to anxiety disorder were screened out through GeneCards， therapeutic target database （TTD）， online mendelian inheritance in man database （OMIM）， and DrugBank， and the drug-disease intersection targets were obtained by taking intersections with the drug targets. The protein-protein interaction （PPI） network was constructed by the STRING database， and the core targets were screened out based on topological parameter analysis. Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were carried out for the intersection targets through the Metascape platform. Maternal separation combined with restraint stress was used to induce the mouse model of anxiety disorder. From the end of lactation on the 21st postnatal day （PD21） to the completion of restraint stress on the 97th postnatal day （PD97）， the mice were fed with Erxian decoction mixed with diet. The anxiety state of mice was evaluated by open field test and elevated O-maze test. The content of plasma corticosterone （CORT） in mice was detected by enzyme-linked immunosorbent assay （ELISA）. The expression levels of protein kinase B （Akt1）， mammalian target of rapamycin （mTOR）， brain-derived neurotrophic factor （BDNF）， postsynaptic density-95 （PSD95）， and synaptophysin in the hippocampus of mice were detected by Western blot and real-time quantitative polymerase chain reaction （Real-time PCR）.ResultNinty-seven active components and 227 action targets of Erxian decoction were obtained. There were 3 863 targets related to anxiety disorder， with 161 drug-disease intersection targets. Among these intersection targets， core targets such as Akt1， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor （TNF）， and mTOR were presumedly closely related to anxiety disorder. The results of KEGG pathway analysis showed that Erxian decoction mainly treated anxiety disorder through phosphatidylinositol 3-kinase （PI3K）/Akt， mitogen-activated protein kinase （MAPK）， and neuroactive ligand-receptor interaction signaling pathways. The results of animal experiments showed that compared with the model group， the Erxian decoction group significantly increased the time of mice spent in the central zone and central crossing times and time spent in the opened arm and opened arm crossing times， with significantly increased expression levels of p-Akt1， p-mTOR， BDNF， PSD95， and synaptophysin （Syp）.ConclusionErxian decoction has the multi-target and multi-pathway characteristics in the treatment of anxiety disorder， and its mechanism may be related to the improvement of synaptic plasticity and neuroinflammation by affecting Akt1， IL-1β， IL-6， TNF， mTOR， and other core targets and modulating PI3K/Akt， MAPK， as well as neuroactive ligand-receptor interaction signal pathways.
Abstract：ObjectiveTo explore the effective components and mechanism of Epimedii Folium in the treatment of oligoasthenotspermia by using network pharmacology and molecular docking technique.MethodThe main active components and corresponding target genes of Epimedii Folium were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. Target genes of oligospermia were obtained by GeneCards and Online Mendelian Inheritance in Man （OMIM） database. Uniprot was used to correct all genes. The drug-active component-key target regulatory network was constructed by Cytoscape3.9.0， and the key active components were screened out according to the degree value. The active components and common targets of the disease were uploaded to STRING 11.5 database to construct the Epimedii Folium and oligoasthenotspermia target protein-protein interaction （PPI） network， and the key protein targets were screened out according to the degree value. The key targets of gene ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed using DAVID database. Protein Data Bank （PDB） and TCMSP were used to obtain the molecular structure of target proteins and active components. AutoDock Vina 1.1.2 was used to perform molecular docking of the active components and the core protein targets. Finally， icariin， the active component of Epimedii Folium， was used to intervene in the rat model of oligoasthenotspermia to verify the effect of icariin on the expression level of protein targets.ResultTwenty-three active components from Epimedii Folium were screened out， and 50 common targets and 6 core targets of oligoasthenotspermia and Epimedii Folium were obtained， including tumor protein p53 （TP53）， epidermal growth factor receptor （EGFR）， prostaglandin-endoperoxide synthase 2 （PTGS2）， cysteine aspartate-specific protease （Caspase）-3， erb-b2 receptor tyrosine kinase 2 （ERBB2）， and caspase-9. Through GO enrichment and KEGG pathway enrichment analysis， the active components of Epimedii Folium were mainly involved in the P53 signaling pathway， the pathways in cancer， cell proliferation， and apoptosis， etc. Molecular docking results indicated that icariin， quercetin， and 8-isopentenol had strong binding ability to target protein. The results of icariin intervention experiment showed that as compared with the control group， the expression of target proteins in testis of rats with oligoasthenotspermia was significantly down-regulated. As compared with the model group， icariin significantly up-regulated the expression of target protein in testis of rats with oligoasthenotspermia （P<0.05）.ConclusionEpimedii Folium treats oligoasthenotspermia through regulating the P53 signaling pathway， the pathways in cancer， cell proliferation， and apoptosis by icariin， quercetin， and 8-isopentenol.
Abstract：ObjectiveTo study the correlations of the characteristics of kidney Yang deficiency syndrome in patients with chronic kidney disease （CKD） with clinical indicators and to explore the risk factors of kidney Yang deficiency in CKD.MethodThe differentiation of traditional Chinese medicine （TCM） syndrome classified the 225 CKD patients who met the inclusion criteria into two groups： one group of kidney Yang deficiency syndrome （99 patients） and one group of non-kidney Yang deficiency syndrome （126 patients）. The symptoms， tongue manifestation， pulse manifestation， and accompanied symptoms of the kidney Yang deficiency syndrome group were recorded. The syndrome characteristics were summarized by factor analysis and clustering analysis. The levels of hemoglobin, red blood cell count, urinary protein, urinary glucose, creatinine, urea nitrogen and glomerular filtration rate were compared between the kidney Yang deficiency syndrome group, the non-kidney Yang deficiency syndrome group and the normal control group by ANOVA and non-parametric test. The binary logistic regression model was employed to analyze the correlations of lifestyle， body mass index （BMI） with syndrome.ResultThe high-frequency symptoms of CKD patients with kidney Yang deficiency syndrome were waist pain， fear of cold， favor of warm， lethargy， fear of cold at waist and knees， etc. The patients mainly presented deep pulse， thready pulse， or weak pulse， and the tongue with white coating， greasy coating， or thin coating. A total of 13 common factors were obtained， which can be classified into 5 categories. The patients with kidney Yang deficiency syndrome mainly had symptoms in limbs （especially lower limbs）， chest， bladder， fleshy exterior， and stomach， with the main manifestations of deficiency-cold， Qi deficiency， fluid retention， and blood stasis. The clustering analysis classified the patients into 11 categories， which reflected that kidney Yang deficiency syndrome mainly presented the symptoms of Qi deficiency， blood stasis， and fluid retention， with fleshy exterior， limbs， spleen， stomach， ears， mind， and bladder involved. The results of clustering analysis and factor analysis were consistent， both of which indicated that the patients were weak with deficiency-cold， accompanied by fluid retention and blood stasis. Frequency analysis also showed that common symptoms mainly included Qi deficiency， fluid retention， cold-dampness， and blood stasis. Compared with the non-kidney Yang deficiency group， the kidney Yang deficiency group showed a large proportion of patients in stage 3-5 CKD， elevated urea nitrogen （P<0.05）， decreased glomerular filtration rate， hemoglobin， and red blood cell count （P<0.05）， and increased qualitative grade of urine protein. In addition， the results of regression analysis showed that female， little or no exercise， and diet preference were the risk factors for kidney Yang deficiency syndrome in CKD （P<0.05）.ConclusionThe disease location and manifestations have correspondence in the CKD patients with kidney Yang deficiency syndrome. The TCM symptoms are correlated with clinical indicators. Hemoglobin， red blood cell count， glomerular filtration rate， urea nitrogen， and urine protein can reflect the connotation of kidney Yang deficiency syndrome in CKD to a certain extent. Additionally， related risk factors in life can affect the occurrence of kidney Yang deficiency syndrome in CKD.
Keywords：chronic kidney disease;kidney Yang deficiency syndrome;syndrome characteristics;data mining;clinical indicators;risk factors
Abstract：ObjectiveTo count and analyze the toxic traditional Chinese medicines and their characteristics in Chinese Materia Medica， so as to provide reference for the development and application of toxic drugs.MethodThe traditional Chinese medicines included in Chinese Materia Medica were screened one by one， and the inclusion criteria were "drug properties"， "usage and dosage" and "major poison， highly poisonous， poisonous， slightly poisonous， slightly poisonous" appearing in ancient books. Standard toxic traditional Chinese medicines were entered into an excel sheet for statistical analysis.ResultA total of 1 408 toxic Chinese medicines were included. The properties and flavors were mainly cold， bitter， pungent and sweet； the main meridians were liver， lung， spleen and stomach； the root， whole grass and leaves were the most used medicinal parts， and there were many toxic drugs. The pre-treatment methods are mainly sun-dried， fresh， fried， calcined， and sunburned； the efficacy categories are mainly heat-clearing drugs， rheumatism drugs， blood-activating and stasis-removing drugs； oral administration methods are mainly decoctions， pills， and powders ， mainly for external application， dipping， and coating； the dosage for oral administration is mostly 9-15 g， 3-9 g， 3-6 g， and an appropriate amount is mainly for external use.ConclusionThere are many toxic Chinese medicines clearly recorded in Chinese Materia Medica， but only 83 kinds of clearly toxic Chinese medicines are included in Chinese Pharmacopoeia， which need to be further strengthened by experimental observation and clinical data verification. The clinical application of toxic traditional Chinese medicine is mainly based on heat toxin blood syndrome and rheumatic arthralgia， which is closely related to its nature， taste and meridian return. Able to move， has the effect of activating Qi and activating blood， "sweet" can replenish energy and slow down， and has the effect of tonic， alleviation and pain relief， and mostly used for the treatment of heat syndrome， blood syndrome and arthralgia syndrome. However， there are certain limitations in the classification and processing conditions of toxic traditional Chinese medicines， which need to be further improved and scientifically verified.
Keywords：Chinese materia medica;toxic traditional Chinese medicine;sexual taste;application characteristics;existing problems
Abstract：Ovarian cancer is a malignant tumor of female reproductive system with high morbidity and mortality. Currently， ovarian cancer patients are mainly treated by primary debulking surgery combined with taxotere/cyclophosphamide （TC） chemotherapy， with the five-year survival rate of 36%-46%. Chinese medicinal materials play a positive role in preventing the occurrence and development of ovarian cancer via multiple targets. The flavonoid monomers in representative Chinese herbal medicines， such as Epimedii Folium， Scutellariae Radix， Curcumae Longae Rhizoma， Ginkgo Folium， Bupleuri Radix， and Longicerae Japonicae Flos， have been proved to have significant anti-tumor activity and been widely used in the treatment of malignant tumors. We reviewed the relevant literature and summarized that flavonoid monomers can regulate multiple signaling pathways to inhibit cell proliferation， block tumor cell cycle， induce apoptosis and autophagy， reduce the ability of cell invasion and migration， inhibit tumor angiogenesis， and reverse platinum resistance， thereby inhibiting the occurrence and development of ovarian cancer. Such pathways include phosphatidylinositol 3- kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway， Janus kinase （JAK）/signal transducer and activator of transcription 3 （STAT3） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， secreted glycoprotein （Wnt）/β-catenin signaling pathway， Notch signaling pathway， and nuclear factor kappaB （NF-κB） signaling pathway. By reviewing the regulatory effect of flavonoid monomers on the signaling pathways of ovarian cancer， we aim to provide a theoretical basis for the research on the roles of flavonoid monomers in inhibiting the occurrence and development of ovarian cancer.
Keywords：flavonoid monomers of Chinese herbal medicines;ovarian cancer;signaling pathway;mechanism;research progress
Abstract：Pyroptosis is a newly discovered programmed cell death. It is an important natural immune response and has obvious anti-infection function. Studies have shown that pyroptosis plays an important role in the occurrence and development of rheumatoid arthritis. Traditional Chinese medicine（TCM） has unique advantages in the prevention and treatment of rheumatoid arthritis. How to guide TCM to effectively prevent and treat rheumatoid arthritis using pyroptosis theory is a new research hotspot in this field. This paper discussed the overview of pyroptosis theory，its mechanism， signal pathway，and application in the treatment of rheumatoid arthritis as well as the research on the activity of TCM based on pyroptosis theory. It was found that the occurrence of pyroptosis was related to Caspase-1-dependent classical inflammatory body pathway and Caspase-1-independent non classical inflammatory body pathway， and pyroptosis produced distinct regulatory effect on the occurrence，development and treatment of rheumatoid arthritis，which would provide a new strategy for the treatment of rheumatoid arthritis. Additionally，TCM recipes such as Miao ethnomedicine prescription Sidaxue and Duhuo Jishengtang， and a variety of effective components such as punicalagin and paeoniflorin monomer derivatives exerted anti-rheumatic and other biological activities by regulating pyroptosis. This provided a theoretical basis and research ideas for the in-depth study of pyroptosis theory and guiding the prevention and treatment of rheumatoid arthritis with TCM.
Keywords：pyroptosis;traditional Chinese medicine;rheumatoid arthritis;research progress;Duhuo Jishengtang;Caspase
Abstract：Mori Cortex is sweet and pungent in taste， cold in nature， and has the tropism to the lung meridian. It has the functions of purging the lung and relieving asthma and can treat oliguria and edema， being one of the commonly used herbal medicines in clinical practice. The prescriptions with Mori Cortex， such as Sangbaipi Tang， Qingjin Huatanfang， and Qingfei Huatantang， are widely used in clinical practice. The main active components in Mori Cortex are the material basis for its efficacy. Owing to the mature methods for the identification of pharmacodynamic substances in Chinese herbal medicines， the research on the chemical components of Mori Cortex has been in-depth and systematic. This article reviews the recent studies about the chemical components and pharmacological effects of Mori Cortex， as well as the treatment of respiratory diseases by the prescriptions with Mori Cortex. On this basis， the effect and mechanism of Mori Cortex and related prescriptions in the treatment of respiratory diseases are summarized. Furthermore， this article analyzes the formulation compatibility and commonly used dosages of Mori Cortex-related prescriptions in clinical practice. It provides reference for the clinical application of Mori Cortex and related prescriptions in the treatment of respiratory diseases.
Abstract：Mitochondrial autophagy is a process to clear dysfunctional mitochondria in the cytoplasm to maintain the integrity of mitochondrial function and cell homeostasis. Mitochondrial autophagy is a complex physiological process， which can maintain the balance of mitochondrial quality and quantity， cell survival under starvation and harsh conditions， and the stability of the intracellular environment. Its molecular mechanism involves a variety of proteins. Many factors can induce mitochondrial autophagy， such as starvation， oxidative stress， hypoxia， depolarization， and other stresses. The accumulation of unfolded proteins can also induce mitochondrial autophagy. In recent years， as a research hotspot， the abnormality of mitochondrial morphology and function is closely related to the occurrence of a variety of diseases. The research on mitochondrial autophagy and the pathogenesis of clinical diseases has attracted more attention， such as tumors， cardiovascular diseases， liver diseases， nervous system diseases， and glucose metabolism disorders. It has been found that regulating mitochondrial autophagy may inspire the treatment of some diseases. Meanwhile， clinical researchers have paid more attention to traditional Chinese medicine （TCM）. As revealed by in-depth research， Chinese medicine has a certain value in regulating mitochondrial autophagy. The research on the pathogenesis of mitochondrial autophagy in related diseases and the intervention of Chinese medicine has found that there are many reports on the regulation of mitochondrial autophagy by Chinese medicine in tumors， cardiovascular diseases， and nervous system diseases. However， the mechanism of mitochondrial autophagy， the balance of mitochondrial autophagy， and the difference in the activation or inhibition of mitochondrial autophagy by Chinese medicine remain unclear. The regulation of mitochondrial autophagy has become a new research target strategy of Chinese medicine in the prevention and treatment of diseases. This paper reviewed the available literature in recent years to provide reference materials for the regulation of mitochondrial autophagy by Chinese medicine and ideas for the follow-up research of Chinese medicine in mitochondrial autophagy.
Abstract：Reflux esophagitis （liver-stomach disharmony， Spittoon-Qi interties， Qi and blood stasis syndrome， turbid poison intrinsic） and nonalcoholic fatty liver disease （liver depression， spleen deficiency， phlegm and blood stasis syndrome， hot and humid embodiment， phlegmy wet resistance） and functional dyspepsia （liver depression syndrome， liver stomach with spleen deficient， spleen deficiency cold syndrome， in a word， fever） is a common disease and frequently encountered disease of digestive system. The course of disease is prolonged and the prevalence is high. The successful establishment of animal model combining disease and syndrome is the premise of exploring the mechanism of traditional Chinese medicine（TCM） effect and the foundation of the development of new preparations. At the same time， mastering the complex relationship network among disease， syndrome and prescription is the prerequisite of effective treatment. When the same syndrome occurs between different diseases， the concept of "treating different diseases with the same treatment" in TCM suggests that methods can be cross-referenced for the shortage of some syndrome models. TCM intervention of digestive diseases has the characteristics of multi-path， multi-target， multi-dimension and multi-level. Therefore， this article through the literature review， summarizes the reflux esophagitis， nonalcoholic fatty liver disease and functional dyspepsia is a common disease such as the combined operation method of the model and the intervention mechanism of TCM， so as to diseases of the digestive system of different syndrome types provides the theory basis for the objective of research， and the basic research of TCM prescription and achievements provide methodological guidance.
Keywords：combination of disease and syndrome;animal model;reflux esophagitis;nonalcoholic fatty liver disease;functional dyspepsia
Abstract：The depressive state， as an emotional disorder， has common symptoms of lack of interest， self-denial， unresponsiveness， unwillingness to communicate， and even extreme misanthropy or suicidal tendency. The depressive state involves a variety of diseases， such as depression， post-stroke depression， postpartum depression， irritable bowel syndrome， major depression， and schizophrenia， which affects the treatment effect and prognosis of the disease， seriously reduces the quality of life and increases the economic burden of patients. At present， the mechanism of depressive state is complex， and the pathophysiological mechanism is unclear. The mechanism of depressive state may be related to abnormal expression of monoamine neurotransmitters， neuronal damage， changes in transduction pathways， hyperactivity of the hypothalamic-pituitary-adrenal （HPA） axis， release of inflammatory cytokines， etc. An increasing number of studies in recent years have confirmed that the occurrence of depression is closely related to intestinal flora disorder， and they interact with each other. Traditional Chinese medicine （TCM） is effective in preventing and treating depressive state with few adverse reactions and a low recurrence rate. TCM also has the effect of regulating the homeostasis of intestinal flora. At the same time， intestinal flora affects the absorption and efficacy of active components of Chinese medicine through metabolic transformation. With the development of microecology， in-depth studies are conducted on the effect of intestinal flora on the occurrence and development of depressive state and brain-gut axis. Intestinal flora has become another potential target for the study of TCM treating depressive state. Starting from the theory and clinical practice of TCM， this paper summarized the mechanism of TCM in treating depressive state by Chinese medicine monomers， compound prescriptions， and acupuncture based on the theory of intestinal flora in recent years. This paper provided information for the profound study of the pathogenesis of depressive state and the scientific connotation of TCM in treating depressive state and ideas for the systematic exploration of the microbiological basis of symptom changes.
Keywords：intestinal flora;depressive state;traditional Chinese medicine;same treatment for different diseases;mechanism
Abstract：Renal interstitial fibrosis（RIF）is a common pathway for the progression of chronic kidney disease to renal failure，and its pathogenesis is mainly related to renal inflammatory damage，oxidative stress，apoptosis，and excessive extracellular matrix（ECM） deposition. Transforming growth factor-β1（TGF-β1） signaling pathway，mammalian target of rapamycin（mTOR） signaling pathway and other signaling pathways mediate the occurrence and development of RIF. Because of the complicated mechanism of RIF，there have been no specific prevention and treatment measures in clinical practice. Autophagy is a non-damaging response produced by eukaryotic cells. It maintains the balance of tissue homeostasis through degradation and reabsorption. At present， Chinese medicine has achieved desirable clinical effects with its unique advantages of multiple components，multiple effects，and multiple targets in the treatment of chronic kidney disease to delay the process of RIF. Scholars have found that autophagy is consistent with the Yin-Yang theory and the theory of abdominal mass in traditional Chinese medicine （TCM） to a certain extent，and it is involved in many aspects of RIF. The progression of RIF is closely related to autophagy. The targeted therapy of RIF by intervention in autophagy has become the frontier of research. However，little is known about the role of autophagy in RIF and the regulation of autophagy by Chinese medicine in the treatment of RIF. Therefore，it is necessary to further elucidate the relationship between autophagy and RIF in order to clarify the mechanism of autophagy in RIF and the mechanism of Chinese medicine regulating autophagy in targeted therapy of RIF. This article focused on the correlation between autophagy and RIF based on TCM theory，and systematically summarized the role of autophagy in RIF and the intervention of Chinese medicine by combining the effects of autophagy on inflammation damage，oxidative stress，apoptosis，and excessive ECM deposition in RIF， and the regulation mechanism of autophagy in TGF-β1 and mTOR signaling pathways in RIF. This study was expected to provide a certain reference for the clinical treatment of RIF and the development of new drugs.