Mechanism of Modified Zhenwutang in Delaying Progression of Chronic Renal Failure Based on Network Pharmacology， Molecular Docking， and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo preliminarily predict the active ingredients， targets， and signaling pathways of modified Zhenwutang in the treatment of chronic renal failure （CRF） based on network pharmacology and explore its potential mechanism for delaying disease progression through molecular docking and animal experiments. MethodThe effective ingredients and targets of modified Zhenwutang were obtained from the HERB database. The targets related to CRF were obtained from the GeneCards. The intersection target genes were obtained using Venny 2.1 software and a protein-protein interaction （PPI） network was constructed using the STRING. The core targets for treating CRF with modified Zhenwutang were screened using Cytoscape 3.9.1 software. The intersection genes were analyzed using Metascape database for gene ontology （GO） analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis. Molecular docking validation was performed using AutoDockTools 1.5.6 software for the key targets and active ingredients. An experimental CRF model was established in rats by administering adenine via gavage for 12 weeks， followed by intervention with modified Zhenwutang and benazepril hydrochloride for four weeks. After treatment， the rats were euthanized， and immunohistochemistry （IHC）， immunofluorescence （IF）， real-time quantitative polymerase chain reaction （Real-time PCR）， and western blot were performed to detect the expression levels of prolyl hydroxylase domain-containing proteins 1 （PHD1）， prolyl hydroxylase domain-containing proteins 2 （PHD2）， hypoxia-inducible factor-1α （HIF-1α）， and α-smooth muscle actin （α-SMA） in the renal tissues of the rats. ResultA total of 426 drug target genes of modified Zhenwutang were obtained from the HERB database. A total of 2 698 target genes related to CRF were obtained from the GeneCards database. There were 154 intersection genes between the drug and the disease. Eight core targets were identified， including albumin （ALB）， protein kinase B1 （Akt1）， tumor necrosis factor （TNF）， interleukin-6 （IL-6）， insulin （INS）， vascular endothelial growth factor A （VEGFA）， tumor protein p53 （TP53）， and interleukin-1β （IL-1β）， which might be closely related to the treatment of CRF with modified Zhenwutang. KEGG enrichment analysis predicted that the main mechanism of modified Zhenwutang in treating CRF involved lipid and atherosclerosis， HIF-1 signaling pathway， cell apoptosis， and nuclear factor kappa B （NF-κB） signaling pathway. Molecular docking results showed that the ingredients of modified Zhenwutang had stable binding activity with the core targets ALB， Akt1， TNF， IL-6， INS， VEGFA， TP53， and IL-1β， which may regulate inflammation and cell apoptosis by affecting the target proteins. The animal model validation results demonstrated that modified Zhenwutang could reduce the expression levels of HIF-1α and α-SMA in the renal tissues of CRF rats， increase the expression levels of PHD1 and PHD2， alleviate renal tissue hypoxia injury， reduce myofibroblast formation， and slow down the progression of CRF in rats. ConclusionModified Zhenwutang may improve renal tissue hypoxia， inhibit cell transdifferentiation， cell apoptosis/necroptosis， and inflammation by affecting the expression of target proteins such as ALB， Akt1， TNF， IL-6， INS， VEGFA， TP53， and IL-1β， as well as regulating the HIF-1 signaling pathway， thus delaying the progression of CRF.

Effect of Mankuining Formula on NLRP3/Caspase-1/GSDMD Pyroptosis Pathway and Gut Microbiota in DSS-induced Ulcerative Colitis Mice / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect of Mankuining Formula （MKNF） on the gut microbiota and the NOD-like receptor （NLR）P3/Caspase-1/gasdermin D （GSDMD） pyroptosis pathway-mediated inflammation in dextran sulfate sodium （DSS）-induced ulcerative colitis （UC） mice. MethodSixty SPF C57BL/6 mice were randomly divided into a blank group， a model group， a MKNF group （20 g·kg-1）， and a mesalazine group （0.266 g·kg-1）， with 15 mice in each group. The UC model was induced in mice by freely drinking a 3% DSS solution for 7 days. After 12 hours of modeling， the treatment groups received daily oral administration， while the other groups received an equal volume of normal saline by gavage. Daily body weight and disease activity index （DAI） were recorded. On the 8th day， mice were euthanized after anesthesia， and the colon and feces were collected. The colon length was measured， and histopathological changes were observed after hematoxylin-eosin （HE） staining. Tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18） levels in the colon were detected by enzyme-linked immunosorbent assay （ELISA）. The differences in gut microbiota among the groups were analyzed using 16S rRNA sequencing technology. The protein content of NLRP3/Caspase-1/GSDMD in colon tissues was detected by Western blot. ResultCompared with the blank group， mice in the model group showed increased DAI （P<0.01）， shortened colon length （P<0.01）， severe colon mucosal damage， elevated levels of TNF-α， IL-1β， and IL-18 （P<0.01）， increased protein content of NLRP3/Caspase-1/GSDMD in colon tissues （P<0.01）， altered gut microbiota structure with decreased abundance of Actinobacteria， Bacteroidetes， and Proteobacteria， and increased abundance of Firmicutes at the phylum level. At the genus level， there was a decrease in Lactobacillus， Alloprevotella， and Yersinia， and an increase in Bacteroides， Bacillus， and Lachnospiraceae_NK4A136. Compared with the model group， the MKNF group and the mesalazine group showed a significant reduction in DAI after the 3rd day （P<0.01）， a significant increase in colon length （P<0.01）， alleviated colon inflammation and mucosal structural damage， and decreased TNF-α， IL-1β， and IL-18 levels in the colon （P<0.01）， reduced protein content of NLRP3/caspase-1/GSDMD in colon tissue （P<0.05， P<0.01），an increase in the abundance of Proteobacteria and Bacteroidetes， and a decrease in Firmicutes at the phylum level. ConclusionMKNF can alleviate UC-induced colonic inflammation， reduce colon damage， and improve dysbiosis of the gut microbiota by inhibiting the classical pyroptosis pathway.

Chrysin alleviates cerebral ischemia-reperfusion injury by inhibiting ferroptosis in rats / 中国中药杂志

The purpose of this study is to investigate whether chrysin reduces cerebral ischemia-reperfusion injury(CIRI) by inhi-biting ferroptosis in rats. Male SD rats were randomly divided into a sham group, a model group, high-, medium-, and low-dose chrysin groups(200, 100, and 50 mg·kg~(-1)), and a positive drug group(Ginaton, 21.6 mg·kg~(-1)). The CIRI model was induced in rats by transient middle cerebral artery occlusion(tMCAO). The indexes were evaluated and the samples were taken 24 h after the operation. The neurological deficit score was used to detect neurological function. The 2,3,5-triphenyl tetrazolium chloride(TTC) staining was used to detect the cerebral infarction area. Hematoxylin-eosin(HE) staining and Nissl staining were used to observe the morphological structure of brain tissues. Prussian blue staining was used to observe the iron accumulation in the brain. Total iron, lipid pero-xide, and malondialdehyde in serum and brain tissues were detected by biochemical reagents. Real-time quantitative polymerase chain reaction(RT-qPCR), immunohistochemistry, and Western blot were used to detect mRNA and protein expression of solute carrier fa-mily 7 member 11(SLC7A11), transferrin receptor 1(TFR1), glutathione peroxidase 4(GPX4), acyl-CoA synthetase long chain family member 4(ACSL4), and prostaglandin-endoperoxide synthase 2(PTGS2) in brain tissues. Compared with the model group, the groups with drug intervention showed restored neurological function, decreased cerebral infarction rate, and alleviated pathological changes. The low-dose chrysin group was selected as the optimal dosing group. Compared with the model group, the chrysin groups showed reduced content of total iron, lipid peroxide, and malondialdehyde in brain tissues and serum, increased mRNA and protein expression levels of SLC7A11 and GPX4, and decreased mRNA and protein expression levels of TFR1, PTGS2, and ACSL4. Chrysin may regulate iron metabolism via regulating the related targets of ferroptosis and inhibit neuronal ferroptosis induced by CIRI.

Tangbikang Granules Modulates AMPK/NF-κB Pathway to Alleviate Sciatic Nerve Inflammation in Diabetic Rats / 中国实验方剂学杂志

ObjectiveTo explore the effect of Tangbikang granules （TBK） on sciatic nerve inflammation in diabetic rats through modulation of adenosine monophosphate-activated protein kinase （AMPK）/nuclear factor （NF）-κB pathway. MethodSD rats were fed with high-fat and high-sugar diet for 8 weeks and then treated with streptozotocin （STZ， ip） at 35 mg·kg-1 for modeling. Then the rats were randomized into diabetes group， low-dose （0.625 g·kg-1）， medium-dose （1.25 g·kg-1）， and high-dose （2.5 g·kg-1） TBK groups， and lipoic acid group （0.026 8 g·kg-1） according to body weight and blood glucose level， and a normal group was designed. After modeling， administration began and lasted 12 weeks. The body mass， blood glucose level， and thermal withdrawal latency （TWL） of the rats were detected before treatment and at the 4th， 8th， and 12th week of administration. At the 12th week， the sciatic nerve was collected for hematoxylin-eosin （HE） and Luxol fast blue （LFB） staining， and the structural changes of sciatic nerve were observed under scanning electron microscope. The levels of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in sciatic nerve were measured by enzyme-linked immunosorbent assay （ELISA）， and the levels of AMPK， phosphorylated （p）-AMPK， and NF-κB proteins in the sciatic nerve were measured by Western blot. ResultThe blood glucose concentration and TWL in the model group were higher than those in the normal group at each time point （P<0.01）. The levels of IL-1β， TNF-α， and NF-κB protein in sciatic nerve in the model group were higher than those in the normal group （P<0.01）， and the p-AMPK/AMPK ratio was smaller than that in the normal group （P<0.01）. Compared with the model group， TBK of the three doses lowered the TWL （P<0.05， P<0.01） and the levels of IL-1β， TNF-α， and NF-κB protein in sciatic nerve of rats （P<0.05， P<0.01）， and high-dose and medium-dose TBK raised p-AMPK/AMPK （P<0.05， P<0.01）. The sciatic nerve fibers were orderly and compact with alleviation of demyelination in rats treated with TBK compared with those in the model group. ConclusionTBK improves the function of sciatic nerve and alleviates neuroinflammation in diabetic rats. The mechanism is the likelihood that it up-regulates the expression of AMPK in the AMPK/NF-κB pathway and inhibits the expression of downstream NF-κB， thereby alleviating the neuroinflammation caused by high levels of inflammatory factors such as IL-1β and TNF-α due to NF-κB activation.

Molecular Mechanism of Tangbikang Granules Against Diabetic Peripheral Neuropathy： Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Tangbikang granules （TBK） against diabetic peripheral neuropathy （DPN） based on network pharmacology and in-vivo experiment. MethodThe active components in medicinals of TBK and their target genes were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. The active components of the medicinals which are not included in TCMSP were searched from previous research. After the analysis of drug-likeness by SwissADME， the target genes of them were predicted with SwissTargetPrediction. DPN-related target genes were retrieved from GeneCards. The common targets of the disease and the prescription were the hub genes of TBK against DPN， which were uploaded to Metascape for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. High-sugar and high-fat diet and low-dose streptozotocin （STZ， ip） were employed to induce diabetes in rats， and then the model rats were respectively treated with low-dose （0.625 g·kg-1）， medium-dose （1.25 g·kg-1）， and high-dose （2.5 g·kg-1） TBK for 12 weeks. Sensory nerve conduction velocity （SNCV） was evaluated. After hematoxylin and eosin （HE） staining， the sciatic nerve was observed under light microscope to examine the nerve damage. Real-time PCR was performed to detect the gene expression of adenosine monophosphate-activated protein kinase （AMPK） pathway-related targets in rat sciatic nerve， and Western blot to measure the protein expression of AMPK and phosphorylated （p）-AMPK in rat sciatic nerve. ResultThe main active components of TBK， such as quercetin， kaempferol， β-sitosterol， leech pteridine A， stigmasterol， and baicalein were screened out， mainly acting on interleukin-6 （IL-6）， tumor necrosis factor （TNF）， protein kinase B （Akt）， JUN， and HSP90AA1 and signaling pathways such as AMPK， nuclear factor-κB （NF-κB）， and Janus kinase/signal transducer and activator of transcription （JAK/STAT）. Molecular docking results showed that β-sitosterol and stigmasterol had high binding affinity with IL-6， TNF， JUN， and HSP90AA1. As for the animal experiment， compared with the normal group， model group had low SNCV of sciatic nerve （P<0.01）， disordered and loose myelinated nerve fibers with axonotmesis and demyelinization， low mRNA expression of AMPKα， AMPKβ， peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α）， Sirtuin 3 （SirT3）， mitochondrial transcription factor A （TFAM）， and low p-AMPK/AMPK ratio in sciatic nerve （P<0.05， P<0.01）. Compared with the model group， TBK of the three doses raised the SNCV （P<0.01）， restored nerve morphology and nerve compactness， and increased the mRNA expression of AMPKα， AMPKβ， PGC-1α， SirT3， and TFAM （P<0.05， P<0.01）. The ratio of p-AMPK/AMPK in the high-dose and medium-dose TBK groups was higher than that in the model group （P<0.01）， while the protein expression in the low-dose TBK group was insignificantly different from that in the model group. ConclusionTBK exerts therapeutic effect on DPN through multiple pathways and targets. The mechanism is that it activates and regulates AMPK/PGC-1α/SirT3 signaling， which lays a basis for further study of TBK in the treatment of DPN.

Effect of Huanglian Ejiaotang on 5-Hydroxytryptamine System and Gut Microbiota in Sleep-deprived Rats / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Huanglian Ejiaotang in intervening in insomnia based on 5-hydroxytryptamine （5-HT） system and gut microbiota. MethodFifty-five SPF-grade SD rats were randomly divided into normal group， model group， low-， medium-， and high-dose Huanglian Ejiaotang groups （1.925， 3.85， and 7.7 g·kg-1）， and Estazolam group （0.1 mg·kg-1）. Except for those in the normal group， the rats in the other five groups were subjected to sleep deprivation on a narrow platform for 12 hours daily for 21 consecutive days. After 14 days of drug intervention， the sleep， exploratory behavior， and depressive-like behavior of the rats were assessed using the pentobarbital sodium sleep synergistic test， the open field test， and the sugar preference test， respectively. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of 5-HT， 5-hydroxyindoleacetic acid （5-HIAA）， tryptophan hydroxylase （TPH）， and monoamine oxidase-A （MAO-A）. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression of the 5-HT1A receptor （5-HT1AR） and 5-HT2A receptor （5-HT2AR）. Differences in gut microbiota among the groups were assessed using 16S rRNA sequencing， and the correlation between the 5-HT system and microbiota was revealed using redundancy analysis. ResultCompared with the normal group， the model group showed a prolonged sleep latency （P<0.05）， reduced sleep maintenance （P<0.01）， decreased central area activity time in the open field （P<0.01）， and reduced sugar preference rate （P<0.05）. Moreover， the model group also showed decreased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， decreased 5-HIAA/5-HT ratio （P<0.01）， downregulated mRNA expression of 5-HT1AR （P<0.01）， and upregulated mRNA expression of 5-HT2AR （P<0.05）. The proportion of Firmicutes decreased， while that of Bacteroidetes increased， leading to a decreased Firmicutes/Bacteroidetes （F/B） ratio （P<0.05）. Compared with the model group， the high-dose Huanglian Ejiaotang group exhibited a shortened sleep latency （P<0.01）， and increased sleep maintenance （P<0.01）. The low-dose Huanglian Ejiaotang group showed increased central area activity time （P<0.01） and an increased sugar preference rate （P<0.05）. The high-dose Huanglian Ejiaotang group exhibited increased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， increased 5-HIAA/5-HT ratio （P<0.05）， upregulated mRNA expression of 5-HT1AR （P<0.01）， and downregulated mRNA expression of 5-HT2AR （P<0.05）. The low-dose Huanglian Ejiaotang group displayed an increased proportion of Firmicutes and a decreased proportion of Bacteroidetes， resulting in an increased F/B ratio. At the phylum level， 5-HT， 5-HIAA， and MAO-A were positively correlated with Firmicutes and negatively correlated with Bacteroidetes. At the genus level， 5-HT， 5-HIAA， TPH， and MAO-A were negatively correlated with Prevotella and Lactobacillus and positively correlated with Blautia and Bacteroides. ConclusionHuanglian Ejiaotang can improve sleep deprivation-induced insomnia and depressive-like behavior by regulating the activity of the 5-HT system and the composition of gut microbiota.

Exploring the therapeutic mechanism of Longqi Fang for diabetic kidney disease based on network pharmacology and verification in rats / 南方医科大学学报

OBJECTIVE@#To study the therapeutic mechanism of Longqi Fang (LQF) for diabetic kidney disease (DKD) based on GEO database and network pharmacology.@*METHODS@#LQF and DKD targets were obtained using the databases including GEO, TCMSP, CNKI, ChemDraw, and SwissTarget Prediction, and LQF-DKD intersection targets were obtained with VENNY. String was used for protein-protein interaction (PPI) analysis, and R package for KEGG and GO enrichment analysis. Cytoscape 3.7.2 software Network graphs were constructed. The results of network pharmacology analysis were verified in SD rat models of DKD by daily treatment of the rats with LQF at low (1 g/kg), medium (2 g/kg), and high (2 g/kg) doses, and kidney pathology was observed with HE staining and the changes in renal function were assessed. Western blotting was used to detect the expression levels of NF-κB and p-NF-κB proteins.@*RESULTS@#We identified 760 main targets of LQF, and obtained 1026 differential genes using GEO database and 61 LQF-DKD intersection targets using Venny database. The core targets obtained through PPI network analysis included Myc, EGF, CASP3, VEGFA, CCL2, SPP1, VCAM1 and ICAM1. Go analysis showed that LQF affects mainly nuclear receptor activity and ligand activated transcription factor activity. KEGG analysis showed that LQF affects inflammatory signaling pathways by interfering with NF-κB, TNF, and PI3K-AKT. In rat models of DKD, treatment with LQF resulted in significant improvements of the renal functions (P < 0.05) and glomerular and tubular structure and arrangement in a dose-dependent manner. Western blotting results showed that LQF dose-dependently downregulated NF-κB and p-NF-κB expressions in the rat models.@*CONCLUSION@#The therapeutic mechanism of LQF for DKD involves multiple components, targets and signal pathways that mediate an inhibitory effect on NF-κB signaling pathway to protect the renal function.

Mechanism of Modified Shengjiangsan in Treatment of IgA Nephropathy Based on Network Pharmacology， Molecular Docking， and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo preliminarily predict the active components， targets， and signaling pathways of modified Shengjiangsan in the treatment of immunoglobulin A nephropathy （IgAN） based on network pharmacology， and to explore its underlying mechanism through molecular docking and experimental verification on animals. MethodThe active ingredients and related targets of modified Shengjiangsan were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP）， UniProt， SwissTargetPrediction， and literature review. IgAN-related targets were obtained from GeneCards and Online Mendelian Inheritance in Man （OMIM）. Cytoscape 3.9.0 was used to construct the regulation network of the related targets of Shengjiangsan and IgAN， and the protein-protein interaction （PPI） network was plotted by STRING. The common genes were analyzed for gene ontology （GO） functional annotation and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment by Metascape. Key targets and main active ingredients were selected for molecular docking by AutoDockTools 1.5.6. The experimental model of IgAN was induced by bovine serum albumin（BSA， ig） combined with lipopolysaccharide （LPS， iv） and the complex of CCl4 and castor oil （sc） in rats. The model rats were treated with modified Shengjiangsan and benazepril hydrochloride for four weeks. The rats were sacrificed after drug administration. The levels of transforming growth factor-β1 （TGF-β1） and interleukin-6 （IL-6） in the serum and kidney tissues were detected by enzyme-linked immunosorbent assay（ELISA）， immunohistochemistry， Real-time quantitative polymerase chain reaction （Real-time PCR）， and Western blot. ResultA total of 105 active ingredients were obtained according to oral bioavailability（OB）， drug-likeness（DL）， and literature screening. There were 124 common genes and 59 core targets. Neurotrophic tyrosine receptor kinase 1 （NTRK1）， cullin-3 （CUL3）， tumor protein 53 （TP53）， epidermal growth factor receptor （EGFR）， exportin 1 （XPO1）， and other targets might be closely related to IgAN. As predicted by KEGG enrichment analysis， the treatment of IgAN with modified Shengjiangsan mainly involved the phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， nuclear transcription factor-kappa B （NF-κB） signaling pathway， and cytokine-cytokine receptor interaction signaling pathway. As revealed by molecular docking， the main active ingredients in modified Shengjiangsan showed stable binding activities with NTRK1， CUL3， TP53， EGFR， and XPO1 in the core targets， indicating that it presumedly regulated inflammatory responses by affecting NTRK1， CUL3， TP53， EGFR， and XPO1 target proteins. The results of experimental verification on animals showed that the expression levels of cytokines TGF-β1 and IL-6 in the serum and kidney tissues of IgAN rats were significantly decreased by modified Shengjiangsan， suggesting that Shengjiangsan might inhibit excessive fibrosis， and inflammatory and immune responses by regulating signaling pathways such as cytokine-cytokine receptor interaction， PI3K/Akt， and NF-κB. ConclusionModified Shengjiangsan may treat IgAN through multiple targets and pathways. Its mechanism may be related to the inhibition of excessive fibrosis， and inflammatory and immune responses by affecting the expression of NTRK1， CUL3， TP53， EGFR， and XPO1 and the regulation of the cytokine-cytokine receptor interaction， PI3K/Akt， NF-κB， and other signaling pathways.

Molecular Mechanism of Erxian Decoction in Treatment of Anxiety Disorder Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

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.

Molecular Mechanism of Erxian Decoction in Treatment of Anxiety Disorder Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

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.

Preliminary study on polymerized human cord hemoglobin increasing the chemosensitivity of subcutaneous xenografts of breast cancer in nude mice / 中国输血杂志

【Objective】 To investigate the effect of polymerized human cord hemoglobin (PolyCHb) on the chemosensitivity of human breast cancer MCF-7 cell subcutaneous xenografts in nude mice and its mechanism. 【Methods】 The MCF-7 cells in exponential growth phase were collected and made into suspension cells at a density of 5×107 cells/mL.Subsequently, the cells were inoculated subcutaneously in the right limb of 18 BALB/c-nu nude mice with 0.2 mL cells per mouse to establish subcutaneous xenograft.When the tumor volume reached about 100 mm3, they were randomly divided into chemotherapy group: doxorubicin 5 mg·kg-1, once/week; chemotherapy + PolyCHb group: in addition to doxorubicin (chemotherapy group), PolyCHb 600 mg·kg-1, 3 times/week; the control group: normal saline 90 mg·kg-1, once/week; all were injected through tail vein continuously for 4 weeks.From the day of injection (d 0), the tumor volume of each group of nude mice was measured every 3 days, and the tumor growth curves were drawn accordingly.After 38 days, the tumor growth observation was completed.The tumor was removed and weighed to calculate the tumor inhibition rate.HE staining, immunohistochemistry and TUNEL method were used to observe the pathological changes of tumor tissue, detect the expression of HIF-1α, and detect tumor cell apoptosis respectively.The content of reactive oxygen species (ROS) of each group was determined by fluorescence staining. 【Results】 The tumor volume (mm3) of chemotherapy + PolyCHb group, chemotherapy group and the control group at day 38 were 196.35±103.45 vs 316.29±62.88 vs 519.42±177.33 (P<0.05), and the tumor inhibition rate (%) of chemotherapy + PolyCHb treatment group and chemotherapy group was 62.20 vs 39.11, respectively.HE staining and TUNEL detection showed that cell necrosis and apoptosis in the growth area of tumor tissue increased in chemotherapy + PolyCHb group.Immunohistochemistry and fluorescence staining showed that HIF-1α expression in chemotherapy + PolyCHb group decreased and reactive oxygen species (ROS) content increased. 【Conclusion】 PolyCHb increases the chemosensitivity of subcutaneous xenograft in nude mice with breast cancer, and its mechanism may be related to the increase of ROS in tumor tissue and the promotion of tumor cell apoptosis.

Literature Study on Current Status of Traditional Chinese Medicine for Chronic Atrophic Gastritis Based on PubMed and Web of Science Databases / 中国实验方剂学杂志

Objective:To review and summarize the current research status of traditional Chinese medicine（TCM） for the treatment of chronic atrophic gastritis（CAG），provide references and hints for relevant studies，and contribute to the further understanding of TCM and the application of TCM in the treatment of CAG with scientific evidence. Method:The PubMed and Web of Science databases were searched for relevant literature on the treatment of CAG with TCM from their establishment to August 31，2020. Eligible randomized controlled trials （RCTs） and animal studies were included according to the inclusion and exclusion criteria，and then the information of the included studies was extracted，summarized，and organized for further analysis. Result:A total of 4 RCTs and 21 animal studies （including 13 papers on compound studies，3 papers on single herb studies，and 5 papers on monomer studies） about TCM treatment for CAG were included in this study. RCTs showed that TCM could work well in improving the pathological state of gastric mucosa and clinical symptoms in patients. However，there were problems of low study quality，and non-uniform diagnostic criteria for gastric mucosal pathology and clinical efficiency evaluation. Animal experiments mainly focused on the study of drug mechanism exploration，and their results showed that TCM treatment of CAG was characterized by multi-target action. However，the animal experiments also had some problems such as inconsistence of CAG animal model establishment，positive drug selection，drug intervention methods as well as intervention cycles among different experiments. Conclusion:The efficacy of TCM in the treatment of CAG has gradually gained global recognition，but there is still a need for further standardization and unification of research methods. In the future，high-quality clinical trials and standardized animal experiments are still needed to conduct in-depth studies on the time for intervention，intervention methods，active ingredients and mechanisms of TCM，so as to make contributions to the full understanding and application of TCM in the treatment of CAG.

Heterotopic ossification after acetabular fracture fixation: Risk factors, prevention and treatment progress / 中国组织工程研究

BACKGROUND: Heterotopic ossification is a common complication of acetabular fracture. Its pathogenesis is still not completely clear, and it is mainly caused by skeletal muscle trauma, hip surgery or nervous system injury. From the research in the past 10 years, the risk factors for ectopic ossification after acetabular fracture surgery are numerous, but its prevention and treatment are controversial. OBJECTIVE: To summarize the relevant literature on the diagnosis, prevention and treatment of ectopic ossification after acetabular fracture surgery in recent years and the risk factors that may affect the prognosis, so as to provide a basis for the clinical prevention and treatment of ectopic ossification after acetabular fracture surgery. METHODS: The databases of Wanfang Medical Network, CNKI and PubMed were searched using computer. The key words were “heteropic ossification; acetabular fractures surgery; NSAIDs; risk factors; radiotherapy; treatment” in Chinese and English. According to the inclusion and exclusion criteria, 67 related articles were finally included and summarized. RESULTS AND CONCLUSION: (1) Although the etiology of heterotopic ossification is not fully understood, it is generally accepted that the formation of heterotopic ossification is determined by both systemic factors and local tissue changes. The risk factors of heterotopic ossification are various, and some preventive measures should be taken for high-risk patients. (2) The prevention of heterotopic ossification after acetabular fracture surgery mainly includes drug, radiotherapy and combined prevention, among which drug prevention, especially non-steroidal anti-inflammatory drugs, is still the most economical and effective prevention method. (3) The treatment of heterotopic ossification is mainly ectopic bone resection and hip arthroplasty. The corresponding treatment methods based on the biomolecular mechanism are still in the stage of animal experimental research, but it points out the direction for the treatment of heterotopic ossification. (4) Among the existing treatment methods, surgery is recognized as the most effective treatment.

Research progress on the protective effect of exercise on retinal cells and functions / 国际眼科杂志(Guoji Yanke Zazhi)

@#Physical exercise is a low-cost, easy-to-implement therapeutic intervention. In recent years, it has been found that exercise has a protective effect on retinal diseases such as diabetic retinopathy, age-related macular degeneration and retinitis pigmentosa. The mechanism may be related to enhancing retinal defence against oxidative stress, reducing glutamate secretion and inhibiting retinal cell apoptosis through BDNF/TrkB pathway. This paper reviews the recent literature related to the protective effect of exercise on retina from animal experiments, clinical studies and mechanism investigation.

Knockdown of lncRNA H19 suppresses endometriosis in vivo

The long noncoding RNA (lncRNA) H19 is involved in the pathogenesis of endometriosis by modulating the proliferation and invasion of ectopic endometrial cells in vitro, but related in vivo studies are rare. This study aimed to investigate the role of lncRNA H19 in a nude mouse model of endometriosis. Ectopic endometrial stromal cells (ecESCs) were isolated from ectopic endometrium of patients with endometriosis and infected with lentiviruses expressing short hairpin RNA (shRNA) negative control (LV-NC-shRNA) or lncRNA-H19 shRNA (LV-H19-shRNA). The ecESCs infected with LV-NC-shRNA and LV-H19-shRNA were subcutaneously implanted into forty 6- to 8-week-old female nude mice. The size and weight of the endometriotic implants were measured at 1, 2, 3, and 4 weeks after implantation and compared, and lncRNA H19 levels in endometriotic implants were evaluated using real-time polymerase chain reaction (RT-PCR). All nude mice survived the experimental period, and no significant differences in body weight were observed between the experimental group and the control group. All nude mice developed histologically confirmed subcutaneous endometriotic lesions with glandular structures and stroma after 1 week of implantation. The subcutaneous lesions in the LV-NC-shRNA group after 1, 2, 3, and 4 weeks of implantation were larger than those in the LV-H19-shRNA group, and lncRNA H19 levels in subcutaneous lesions in the LV-NC-shRNA group were significantly higher than those in the LV-H19-shRNA group. Knockdown of lncRNA H19 suppresses endometriosis in vivo. Further study is required to explore the underlying mechanism in the future.

Changes of cardiac function in rats with myocardial infarction after umbilical cord-derived mesenchymal stem cell transplantation: A meta-analysis / 中国组织工程研究

BACKGROUND: It has been shown that compared with stem cells from other sources, umbilical cord-derived mesenchymal stem cells have lower immunogenicity, and their application has significant effect in rats with myocardial infarction. OBJECTIVE: To systematically evaluate the effects of umbilical cord-derived mesenchymal stem cells on cardiac function of rats with myocardial infarction. METHODS: PubMed, Cochrane, Embase, CBM, CNKI, WanFang, VIP and CJD databases were retrieved for the literature concerning umbilical cord-derived mesenchymal stem cells for treating rats with myocardial infarction published before June 2019. Two researchers independently completed literature screening, data extraction and methodological quality evaluation according to the inclusion criteria. Meta-analysis was conducted on Stata 14.0. RESULTS AND CONCLUSION: A total of 9 articles were included, involving 216 rats. Meta-analysis showed that: (1) Umbilical cord-derived mesenchymal stem cells significantly increased the left ventricular ejection fraction after myocardial infarction in rats [95% confidence interval (CI) (3.16, 3.76), P < 0.001]. (2) Umbilical cord-derived mesenchymal stem cells significantly increased the left ventricular short axis shortening rate after myocardial infarction in rats [95%CI (0.18, 0.54), P < 0.001]. (3) Umbilical cord-derived mesenchymal stem cells significantly shortened the left ventricular end-diastolic internal diameter [95%CI (-1.90,-0.99), P=0.042] and left ventricular end-systolic internal diameter [95%CI (-6.56,-4.65), P < 0.001]. (4) Umbilical cord-derived mesenchymal stem cells significantly improved the left ventricular end-diastolic volume [95%CI (-2.01,-1.11), P < 0.001] and left ventricular end-systolic volume [95%CI (-3.44,-2.17), P < 0.001]. In summary, umbilical cord-derived mesenchymal stem cell transplantation is effective and safe in the treatment of myocardial infarction in rats. Due to the limitation of the quality of the included literature, the above conclusions need to be validated by high-quality and large-sample randomized controlled trials.

A questionnaire-based comparison of conventional teaching methods to computer assisted learning in experimental pharmacology for undergraduate medical students

Background: Computer Assisted Learning (CAL) for teaching experimental pharmacology is now widely accepted as alternative to animal experiments. The objective of this study was to compare the performance of undergraduate medical students by conventional teaching methods (lecture and discussion) and CAL.Methods: This was a questionnaire based observational study involving 109 MBBS students of fifth semester. The students were taught experimental pharmacology practical by both conventional methods (lecture with discussion) and computer assisted learning (CAL). Questionnaire and their filled responses by these students were taken at the end of lecture-discussion (pre-CAL) and after the CAL experiments (post-CAL), covering the same topics of experimental pharmacology. Pre-CAL and post-CAL data were assessed statistically.Results: In the pre-CAL session, only 53.39%, 47.56%, 53.39% and 49.5% of the students were having the scores above 40% in the rabbit eye, DRC and potentiation, frog heart and dog BP experiment respectively, which was increased to 77.44%, 75.48%, 75.47% and 75.48% of the students respectively in the post-CAL assessment. A statistically significant difference (p <0.05) in the performance was observed among the students in the pre-CAL and post-CAL assessment.Conclusions: CAL is a good alternative to animal experimentation. Lecture with discussion, followed by CAL experiments on the same topics, enhanced the performance of students as shown by improvement in post-CAL scores.

Systematic review on the animal experiments of electroacupuncture for hyperlipidemia combined with atherosclerosis / 中国针灸

OBJECTIVE@#To systematically review the effectiveness and safety of electroacupuncture (EA) on hyperlipidemia combined with atherosclerosis in animal experiments.@*METHODS@#The relevant animal experiments of EA treatment for hyperlipidemia combined with atherosclerosis were retrieved from the CNKI, WANFANG, VIP, CBM, PubMed, Embase and the Cochrane Library up to March 19, 2018 since the establishment of databases. Two researchers in the project team screened the literature and extracted the information such as experiment objects, experiment steps and experiment results separately. The literature assessment was evaluated. Meta analysis was performed with RevMan 5.3 software.@*RESULTS@#Eventually, 29 eligible experimental researches were included and the average score in the quality assessment was 5.1, at the moderate level. Meta analysis showed that compared with the model control group, after EA treatment, the levels of the total cholesterol (TC) [ -2.29, 95% (-2.94, -1.64), <0.000 01], triglyceride (TG) [ -1.30, 95% (-1.76, -0.84), <0.000 01] and low density lipoprotein-cholesterol (LDL-C) [ -2.64, 95% (-3.29, -1.99), <0.000 01] were significantly reduced and the level of high density lipoprotein-cholesterol (HDL-C) [ 0.55, 95% (0.06, 1.04), =0.03] was increased, indicating the significant differences.@*CONCLUSION@#On the base of the current limited evidences, EA significantly reduces the levels of TC, TG and LDL-C and slightly increases the level of HDL-C in the animal models of hyperlipidemia combined with atherosclerosis.

A survey on the knowledge and perception of computer assisted learning among undergraduate medical students in a tertiary care teaching institute of North India

Background: Computer assisted learning (CAL) in the classroom as well as laboratory in the medical profession has been rising in the present scenario worldwide. CAL can replace laboratory based animal experiments to a large extent and prevent the unnecessary harm or killing of animals. In India, only few studies have been conducted on CAL till date. Aims and objective of the study was to evaluate the knowledge and perception on CAL among undergraduate medical students.Methods: This cross sectional study was carried out on the medical students (MBBS-Second Professional) in the department of pharmacology at Guru Gobind Singh Medical College and Hospital, Faridkot, Punjab, India. Feedback was obtained from 105 students who had already performed animal experiments in CAL laboratory.Results: It was seen that Computer simulated models (CSMs) are useful for undergraduate teaching (by 98.1%), CAL enrich learning experience about drug effects (by 97.1%), CSMs in CAL is user friendly (by 97.1%) and use of CSMs in teaching is ethical (by 97.1%).Conclusions: CAL is an innovative teaching and learning technique for the medical students. This is an effective mode of teaching pharmacology to the students and it also helps the students to apply their theoretical knowledge of drugs to the practical aspects (without use of animal) on computer simulated models.

The effects of different dose of nonionic iodine contrast agent on the renal function of dogs / 实用放射学杂志

Objective To evaluate the change of serum creatinine (Scr) before and after administration of contrast agent in different dose,to observe the difference of dog's kidney tissue with electron microscopy and investigate the effect of contrast agent on renal function.Methods Twelve dogs were divided into four groups randomly:the control group,the low dose group,the moderate dose group and high dose group.After the administration of different doses of iodine contrast agent at the same rate,the changes of Scr and microscopic structure were compared before administration and 48 hours later.Results The differences of Scr before and 48 hours after administration were (4.6±1.6) μmol/L,(6.7±2.5) μmol/L,(6.9±4.5) μmol/L,(5.1± 1.9) μmol/L for control group,low dose group,moderate dose group and high dose group,respectively.There was no statistically significant difference among the groups (P ＞0.05).In high dose group,the mitochondria of tubular epithelial cells were swelling and obvious vacuoles were observed.Only a small amount of vacuoles existed in the renal tubular epithelial cells in low dose group.Conclusion Compared with the moderate and high dose group,the low-dose iodine contrast agent have less damage to the kidney cells of the dogs.