Analysis of in vitro activity and compatibility of Dunhuang Yifang Dabupi Decoction on gastric cancer based on computer-aided drug design / 药学学报

Based on the research system of computer-aided drug design combined with complex network analysis, the potential mechanism of Dunhuang Dabupi Decoction in preventing and treating gastric cancer (GC) is analyzed, and the scientific connotation of its prescription rules is explored through the efficacy grouping. To study the effect of Dabupi Decoction freeze-dried powder solution on the proliferation activity of gastric cancer cells through cell experiments; the TCMSP and TCMID databases were used to collect the compound components of Dabupi Decoction. Swiss Target Prediction is used to predict potential targets of compounds. DrugBank, GeneCards, TTD, and DisGeNET were used to collect potential targets for gastric cancer. Analyze protein interactions of potential targets through the STRING database. DAVID database was used for KEGG enrichment analysis; Dabupi Decoction was divided into Wenzhong group (dried ginger), Yiqi group (ginseng, licorice, Atractylodes macrocephala), nourishing Yin group (Ophiopogon japonicus, Schisandra) and Jiangni group according to its efficacy characteristics. The inverse group (inula) has 4 functional compatibility groups. Cytoscape was used to construct a network of "medicinal flavor-potential active ingredient-key target" respectively, and the network was used to analyze the scientific connotation of the compatibility of efficacy groups. The Schrödinger software was used to verify the molecular docking of the core components and the core targets. The material basis of the Dabupi Decoction to prevent and treat gastric cancer was discovered through the combination of pattern analysis and combined free energy calculation. The core drug was analyzed from the perspective of dynamics through molecular dynamics simulation. Potential targets and representative potential compounds interact with each other. Cell experiments confirmed that Dabupitang freeze-dried powder solution can down-regulate the mitochondrial membrane potential of AGS gastric cancer cells, block the cell cycle in the G0/G1 phase (P < 0.05), and inhibit its proliferation (P < 0.05). The pathways enriched by the four functional groups contained in Dabupi Decoction are mainly distributed in the body's energy metabolism, inflammation-immune system regulation, and cycle-apoptotic functions. Each module is connected by a common target gene and has its own focus. The results of molecular docking showed that the compounds liquiritigenin, quercetin, kaempferol, isorhamnetin, methylophiopogonanone A, etc. may be the effective multi-target components of Dabupi Decoction. Estrogen receptor 1, androgen receptor, ATP-binding cassette superfamily G member 2, epidermal growth factor receptor, glycogen synthase kinase-3 beta and other targets have good affinity with each potential active compound, which may be a potential target of Dabupi Decoction for preventing and treating gastric cancer. Among them, kaempferol and the drug target EGFR not only have good binding ability, but also have good binding stability. This study is based on computer-aided drug design combined with complex network analysis strategies to initially reveal the material basis and molecular mechanism of Dabupi Decoction in the prevention and treatment of gastric cancer. It also explores the scientific connotation of Dabupi Decoction in the prevention and treatment of gastric cancer with different efficacy groups, and its clinical application provide chemical bioinformatics basis.

Material basis and molecular mechanism of Angelicae Sinensis Radix in activating blood:based on computer-aided drug design / 中国中药杂志

Angelicae Sinensis Radix excels in activating blood, but the scientific mechanism has not been systematically analyzed, thus limiting the development of the medicinal. This study employed the computer-aided drug design methods, such as structural similarity-based target reverse prediction, complex network analysis, molecular docking, binding free energy calculation, cluster analysis, and ADMET(absorption, distribution, metabolism, excretion, toxicity) calculation, and enzyme activity assay in vitro, to explore the components and mechanism of Angelicae Sinensis Radix in activating blood. Target reverse prediction and complex network analysis yielded 40 potential anticoagulant targets of the medicinal. Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis indicated that the targets mainly acted on the complement and coagulation cascade signaling pathway to exert the anticoagulant function. Among them, the key enzymes thrombin(THR) and coagulation factor Xa(FXa) in coagulation cascade and thrombosis were the drug targets for thromboembolic diseases. At the same time, molecular docking and cluster analysis showed that the medicinal had high selectivity for FXa. According to binding free energy score, 8 potential active components were selected for enzyme activity assay in vitro. The results demonstrated that 8 components inhibited THR and FXa, and the inhibition was stronger on FXa than on THR. The pharmacophore model of 8 active compounds was constructed, which suggested that the components had the common pharmacophore AAHH. The ADMET calculation result indicated that they had good pharmacokinetic properties and were safe. Based on target reverse prediction, complex network analysis, molecular docking and binding free energy calculation, anticoagulant activity in vitro, spatial binding conformation of molecules and targets, pharmacophore model construction, and ADMET calculation, this study preliminarily clarified the material basis and molecular mechanism of Angelicae Sinensis Radix in activating blood from the perspective of big data, and calculated the pharmacology and toxicology parameters of the active components. Our study, for the first time, revealed that the medicinal had obvious selectivity and pertinence for different coagulation proteins, reflecting the unique effect of different Chinese medicinals and the biological basis. Therefore, this study can provide clues for precision application of Angelicae Sinensis Radix and the development of the blood-activating components with modern technology.

Research progress on Astragali Radix and prediction of its quality markers (Q-markers) / 中国中药杂志

Astragali Radix, a medicinal herb for invigorating Qi, has anti-aging, anti-tumor, immunoregulatory, blood sugar-and lipid-lowering, anti-fibrosis, anti-radiation and other pharmacological effects. This article reviewed the studies about the chemical components and pharmacological effects of Astragali Radix. According to the theory of quality markers(Q-markers) of Chinese medicinal materials, we predicted the Q-markers of Astragali Radix from traditional efficacy, chemical component validity, measurability, plant phylogeny, and pharmacokinetis. The results showed that total polysaccharides, flavonoids(e.g., calycosin-7-O-β-D-glucoside, formononetin, calycosin, quercetin, and ononin), and saponins(e.g., astragalosides Ⅱ, Ⅲ, and Ⅳ) can be taken as the main Q-markers. This review lays a foundation for regulating the quality research and standard establishment of Astragali Radix, and benefits the control and quality supervision of the production process of Astragali Radix and its related products.

Effect of Dunhuang Pingweiwan and Its Decomposed Recipes on Tumor Inhibition and PI3K/Akt/mTOR Pathway in SCG-7901 Gastric Cancer Mice / 中国实验方剂学杂志

Objective:To investigate the antitumor effect and the mechanism of Dunhuang Pingweiwan and its decomposed recipes based on phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway in SCG-7901 gastric cancer-mice. Method:The subcutaneous tumor bearing model of SCG-7901 gastric cancer in mice was established， and the the mice were randomized into model group， Dunhuang Pingweiwan group （14.04 g·kg^-1·d^-1）， Huoxue Jiedu group （6.50 g·kg^-1·d^-1）， Wenzhong Sanhan group （3.64 g·kg^-1·d^-1） and cisplatin group （2 mg·kg^-1·d^-1）， with 8 mice in each group. From the 8^th day of inoculation， the mice were administered for 10 consecutive days. The mice were weighed and the general conditions were observed every other day. On the next day of the last administration， the mice were sacrificed， and the tumor was removed and weighed to calculate the anti-tumor rate. The histopathological changes were observed by hematoxylin-eosin （HE） staining， and the mRNA and protein expressions of PI3K， Akt， and mTOR in tumor tissues were detected by real-time polymerase chain reaction（Real-time PCR） and immunohistochemistry （IHC）， respectively. Result:From the 10^th day of inoculation， the mice in cisplatin group were generally in poor condition and their body mass decreased. The mice in model group， Dunhuang Pingweiwan group， Huoxue Jiedu group and Wenzhong Sanhan group were generally fair， and their body mass increased without significant difference among groups. The tumor inhibition rates of Dunhuang Pingweiwan， Huoxue Jiedu， Wenzhong Sanhan and cisplatin groups were 30.74%， 24.80%， 4.19% and 63.84%， respectively. Except for Wenzhong Sanhan group， tumor weight of the other treatment groups was significantly lower than that of the model group （<italic>P</italic><0.01）， and there was no significant difference between the Dunhuang Pingweiwan and Huoxue Jiedu group. Dunhuang Pingweiwan and Huoxue Jiedu group could significantly reduce tumor cell density and cause tumor cell necrosis. Compared with the model group， the expressions of PI3K， Akt， and mTOR mRNA and protein in the Dunhuang Pingweiwan， Huoxue Jiedu and cisplatin groups significantly decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， and there was no significant difference between the Dunhuang Pingweiwan group and Huoxue Jiedu group. Conclusion:Dunhuang Pingweiwan and its decomposed recipes （Huoxue Jiedu） have a certain anti-tumor effect on the SCG-7901 gastric cancer-mice， and the mechanism may be related to the down-regulation of key molecules in the PI3K/Akt/mTOR signaling pathway.

Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis / 中国实验方剂学杂志

<b>Objective::To observe the effect of Youguiwan on the levels of cartilage transducers and activators of transcription 3 (STAT3) and interleukin-6 (IL-6) in rats with knee osteoarthritis (KOA). <b>Method::Sixty SD rats were randomly divided into six groups: sham control group, model group, glucosamine sulfate group and Youguiwan (high, middle and low-dose) groups. The modified Hulth method was used to prepare KOA models for 6 weeks. The shame control group and the model group were treated with normal saline, Youguiwan high, middle, low-dose groups received Youguiwan 4.8, 2.4, 1.2 g·kg^-1 by gavage respectively, and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g·kg^-1. The rats were administrated for 8 weeks according to the dose. After intervention, each group was put to death by femoral artery blood collection, and the keen cartilages of the rats were collected. The pathological changes were observed by htoxylin eosin (HE) staining method, and Mankin score was evaluated. The expressions of STAT3, superoxide dismutase3(SOD3) and Wnt inhibitory factor 1(WIF1) in articular cartilage were detected by immunohistochemistry. The expressions of IL-6 mRNA in articular cartilage were detected by quantitative real-time fluorescence polymerase chain reaction (Real-time PCR). The expression of WIF1 in articular cartilage was detected by Western blot. <b>Result::Compared with the sham control group, the Makin score was obviously increased in the model group, the protein expression of STAT3 was increased significantly, the mRNA of IL-6 was raised significantly, but the protein expression of WIF1 was decreased significantly (<italic>P</italic><0.01), articular cartilage was seriously damaged, and chondrocytes were arranged in disorder. Compared with the model group, the Makin score was declined obviously in the high-dose Youguiwan group, the protein expression of STAT3 was significantly reduced, the mRNA expression of IL-6 was significantly reduced in Youguiwan treatment group, while the protein expression of WIF1 was significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), the cartilage structure returned to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. <b>Conclusion::Youguiwan could significantly improve the articular cartilage degeneration of KOA rats, and inhibit the inflammation of chondrocytes, which may be related to the suppression of STAT3 and IL-6 expression.

Pharmacological Analysis of Xuanfei Huazhuo Prescription in Treatment of Coronavirus Disease-2019 / 中国实验方剂学杂志

Objective:The targets and signaling pathways of Xuanfei Huazhuo prescription (XFHZP) for the treatment of coronavirus disease-2019 (COVID-19) were explored, and its possible action mechanisms were described through network pharmacology and basic analysis of modern pharmacology. Method:The compounds and targets in XFHZP were collected through TCMSP and BATMAN-TCM databases. The targets of COVID-19 were studied by GeneCards, NCBI and CTD databases. The PPI network was constructed through STRING database. The networks of "herb-meridian" and "traditional Chinese medicines-compounds-targets-disease" were generated by Cytoscape 3.7.0. Then, Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis and Gene Ontology（GO） analysis were made for shared targets through the Omicshare platform. In addition, the disease targets of multiple organ injury, immune injury and severe acute respiratory syndrome (SARS) were retrieved and then mapped with XFHZP. The ratio of intersection targets to XFHZP's targets was calculated. Result:XFHZP has 10 traditional Chinese medicines in total, including 6 medicines with the meridian tropism to lung, 5 medicines with the meridian tropism to the spleen and 5 medicines with the meridian tropism to the stomach. There were 409 compounds and 2 271 targets. There were 8 same inflammatory factors in targets between XFHZP and COVID-19, and each inflammatory factor corresponded to multiple compounds. XFHZP and COVID-19 had 135 intersection targets, and 36 key targets were screened out. A total of 172 signaling pathways were screened out through KEGG signal pathway enrichment (P<0.05). There were 4 000 biological processes, 254 cell components, and 408 molecular functions (P<0.05) according to GO analysis. XFHZP had many common targets with various organ damage targets and immune damage targets, with the ratio of about 7.6%-97.8%. XFHZP had 173 intersection targets with SARS. Conclusion:XFHZP may treat COVID-19 through anti-inflammatory, organ protecting and immune effects. It will provide a certain theoretical basis for the development of drugs for COVID-19.

Material Basis of Xuanfei Huazhuo Prescription in Treatment of COVID-19 by Multi-target Molecular Docking / 中国实验方剂学杂志

Objective:Structure-based angiotension converting enzyme 2 (ACE2) and interleukin-6R (IL-6R) were taken as the target proteins to in the investigation of the material basis of Xuanfei Huazhuo prescription in the treatment of coronavirus disease-2019 (COVID-19) by molecular docking. Method:The compounds in Xuanfei Huazhuo prescription were retrieved through TCMSP. Structure-based ACE2 and IL-6R were taken as the target proteins to screen out the compounds with a better activity by molecular docking, and analyze structural properties of these compounds. Furthermore, the potential molecular mechanism of Xuanfei Huazhuo prescription in the treatment of COVID-19 was analyzed by target reverse prediction. Result:There were 312 potentially active compounds in Xuanfei Huazhuo prescription, including 75 highly active compounds and 15 highly active compounds for ACE2. There were 100 eligible active compounds and 3 highly active compounds for IL-6R, most of which belong to flavonoids. The herb-component-target network included 10 herbs, 126 compounds and 130 targets. String analysis showed that PIK3R1, SRC, AKT1, AR and EGFR might be the key targets of Xuanfei Huazhuo prescription. Conclusion:Based on the virtual screening of multi-target molecular docking, the anti-virus and anti-inflammatory material basis of Xuanfei Huazhuo prescription was preliminarily obtained. At the same time, based on the reverse prediction and analysis, potential targets and molecular mechanism of the recipe in the treatment of COVID-19 were explored, so as to provide clues for the multi-angle mining of Xuanfei Huazhuo prescription and its relevant prescriptions and the modernization development of monomer components.

Clinical Efficacy of Xuanfei Huazhuo Prescription on 40 Cases of COVID-19 / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Xuanfei Huazhuo prescription in the treatment of coronavirus disease-2019 (COVID-19). Method:A total of 40 patients with COVID-19 were selected and treated with Xuanfei Huazhuo prescription. The changes of body temperature, clinical symptoms, computed tomography (CT), blood routine and biochemical indexes were observed before and after treatment. Result:The 40 patients included 15 males and 25 females, with a male to female ratio of 1∶1.7. They were aged between 20-94 years old, with the average age of （43.9±16.3） years old. The course of disease was 8-23 days, with the average of （14±4.4） days. Compared with before administration, the patients' clinical symptoms, such as cough, fever, sputum, diarrhea, loss of appetite and fatigue, were all improved (P<0.05). Before treatment the traditional Chinese medicine (TCM) syndromes of patients were mainly cold dampness lung (57.5%) and cold dampness Lung (42.5%), and the tongue coating was mainly white greasy coating (52.9%). After adjuvant treatment with Xuanfei Huazhuo prescription, the fever removal time was (2.48±2.56) days; white blood cell (WBC), lymphocyte percentage (LYM%), neutrophil percentage (NEUT%), absolute value of lymphocytes (LYM #) indexes of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), total bilirubin (TBIL), ratio of glutamic oxaloacetic transaminase to glutamic pyruvic transaminase (AST/ALT) and lactate dehydrogenase (LDH) were basically restored to the normal range (P<0.05) compared with before administration. After adjuvant treatment with Xuanfei Huazhuo prescription, the results of three pharyngeal test virus nucleic acid tests were negative, and the lung CT showed that infected lesions were absorbed and all met the discharge criteria. All 40 patients met the discharge criteria and were all cured and discharged, with a cure rate of 100%. There has been no case of recurrence with a positive result of nucleic acid detection so far. The score of symptom and clinical index of patients after administration was （1.62±1.90）, which was significantly lower than that before administration (7.65±4.08, P<0.05). Conclusion:In the adjuvant treatment of COVID-19, Xuanfei Huazhuo prescription can reduce body temperature, promote the absorption of pulmonary inflammation, and improve clinical symptoms, such as fever and cough.

Analysis of "Gansu Prescriptions" in Prevention and Treatment of Coronavirus Disease-2019 / 中国实验方剂学杂志

Coronavirus disease-2019 (COVID-19) is an acute infectious disease caused by a 2019 novel coronavirus (2019-nCoV) infection. It is highly contagious, and can spread quickly home and abroad. It has caused a global pandemic. After the outbreak, Gansu province actively responded to the national "integrated Chinese and western medicine(ICWM)" epidemic prevention policy by organizing an expert group on the prevention and treatment of traditional Chinese medicine(TCM) and establishing a joint working mechanism of ICWM. In adherence to the principle of ICWM, it highlighted the advantages of TCM in epidemic prevention, and emphasized early, timely and whole course use of TCM. The expert group continued to summarize in practice and form a series of "Gansu prescriptions", so as to explore the prevention and control strategy of "prevention in advance, timely interruption and reversal, early prevention and cure, and cure in early stage". Before illness, the prevention shall be made in advance by taking Fuzheng Biwen prescription based on constitution differentiation, in order to strengthen the body resistance and removing pathogenic Qi, after the onset, the syndromes were first treated, interrupted and reversed, and Xuanfei Huazhuo prescription and Qingfei Tongluo prescription were administered based on syndrome differentiation, so as to exorcise pathogenic Qi and cure COVID-19 at the early stage, at the beginning stage of recovery, Jianpi Yifei prescription was used to strengthen the spleen and lungs, and harmonize the stomach and resolve dampness, so as to prevent recurrence. In the principle of ICWM, "Gansu prescriptions" were selected based on the constitution differentiation and syndrome differentiation, so as to prevent the occurrence of epidemics, block light and common symptoms from developing to heavy and critical symptoms, improve the clinical efficacy, shorten the course of disease, and reduce the incidence of critical illness, thereby reducing mortality.

Effect of M1/M2 Alveolar Macrophage Subsets in COVID-19 and Regulatory Mechanism of Traditional Chinese Medicine / 中国实验方剂学杂志

The pathological anatomical results of coronavirus disease-2019 (COVID-19) patients showed that excessive inflammatory reaction in the lungs is one of the important causes for such complications as acute lung injury or acute respiratory distress syndrome. Therefore, regulation of immune response may be an effective measure for COVID-19. Alveolar macrophages have a high heterogeneity and plasticity. The dynamic changes of subsets balance and function of M1/M2 alveolar macrophages have a significant effect on pulmonary inflammatory response during the early and late stages of infection. This paper reviews the classification and function of macrophages and explores the mechanism of alveolar macrophage in the pathological process of COVID-19 at different stages and the pharmacodynamic mechanism of traditional Chinese medicine. Besides, it provides ideas for the treatment of COVID-19 with traditional Chinese medicine and other drugs' research and development based on the regulation of macrophage polarization.

Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde / 中国实验方剂学杂志

Objective::To study the protective effect of astragalus polysaccharide (APS) on micronucleus and sister chromatid exchange (SCE) in human bone marrow mesenchyml stem cell (BMSCs) exposed to formaldehyde, in order to initially explore the potential mechanism. Method::BMSCs were cultured in vitro, cells were randomly divided into five groups: control group, formaldehyde group, and APS 40, 100, 400 mg·L-1 groups. BMSCs were infected with 120 μmol·L-1 formaldehyde, meanwhile, APS 40, 100, 400 mg·L-1 groups were co-cultured with 40, 100, 400 mg·L-1 APS. Cell morphology was observed by inverted phase contrast microscope, micronucleus were detected by micronucleus test, SCE was detected by SCE test, and mRNA and protein expressions of proliferating cell nuclear antigen (PCNA), xeroderma pigmentosum B, D, F, G (XPB, XPD, XPF, XPG) were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)and Western blot. Result::Compared with control group, cell counts decreased, and cell morphology of BMSCs in formaldehyde group significantly changed, they were all recovered gradually in 40, 100, 400 mg·L-1 APS groups. Compared with control group, the micronucleus and SCE increased significantly (P<0.01), PCNA mRNA and protein expressions down-regulated significantly (P<0.05), while XPB, XPD, XPF, XPG mRNA and protein expressions up-regulated significantly (P<0.05, P<0.01). Compared with formaldehyde group, BMSCs were treated with APS at 40, 100, 400 mg·L-1, micronucleus and SCE decreased significantly (P<0.01), and mRNA and protein expressions of PCNA, XPB, XPD, XPF and XPG up-regulated significantly (P<0.05, P<0.01). Among them, the 100 mg·L-1 APS group had the most obvious effect. Conclusion::APS can protect formaldehyde-induced BMSCs micronucleus and SCE, especially 100 mg·L-1 APS has the most obvious effect. The mechanism may be associated with the up-regulation of expressions of PCNA, XPB, XPD, XPF and XPG in the nucleotide exicision repair pathway (NER), which promoted the damage repair.

Possile Effect of Small Molecule in Hedysari Radix in Antagonizing Tumor Necrosis Factor Receptor 1 Based on Molecular Docking / 中国实验方剂学杂志

Objective: To study the effect of small molecule compounds of Hedysari Radix in ntagonizing tumor necrosis factor receptor type 1 (TNFR1) based on molecular docking. Method: The structure of small molecular compound of Hedysari Radix was downloaded from the chemical composition compound library of traditional Chinese medicine, and then optimized to obtain the composition compound library of Hedysari Radix. The three-dimensional structure of the inflammatory target TNFR1 (PDB ID:1TNR) was identified. After hydrotreating and anhydrating, the binding pocket residues were identified according to the literature. According to the defined target structure and binding pocket, the flexible molecular docking was conducted between the composition compound library and the target, and the score (Glide Score) was obtained. Based on the results of molecular docking, the first nine small molecular compounds of Glide Score were selected as candidate components. On this basis, the drug-likeness was analyzed, which involved small molecular compounds that meet the number of hydrogen-bonded receptors, the number of hydrogen-bonded donors, the formula weight, the number of rotatable key and the numerical range of lipo-hydro partition coefficient. Finally, the binding mode was analyzed according to pharmacokinetic parameters and complex structure of composition-target docking. Result: The residue set in the TNFR1 drug-binding pocket were identified as glutamic acid109 (Glu109), lysine 35(Lys35), alanine62 (Ala62), serine 74 (Ser74), lysine75 (Lys75), cysteine76 (Cys76), argnine 77(Arg77), glutamine82 (Gln82), threonine89 (Thr89), asparticacid91 (Asp91), argnine92 (Arg92), aspartic acid93 (Asp93), threonine 94(Thr94), valine95 (Val95), cysteine 96(Cys96), argnine104 (Arg104), tyrosine106 (Tyr106), asparagine110 (Asn110), leucine111 (Leu111), phenylalanine112(Phe112), glutamic acid 131(Glu131) and lysine132 (Lys132). Totally 43 small molecular compounds of Hedysari Radix were obtained. Five small molecular compounds, namely hedysari radix, quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin, were screened by comprehensive factors, like docking scoring. Conclusion: Quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin are the effective anti-inflammatory substances of Hedysari Radix, with a great possibility of becoming TNFR1 antagonists.

The research progress of interaction between inflammatory environment and MSCs autophagy / 中国药理学通报

Mesenchymal stem cells (MSCs) can participate in the repair of various tissue and organ damage, but inflammation often exists in local repair area,and autophagy is induced. As a kind of self-regulating mechanism of cells, autophagy can not only regulate the physiological process of MSCs under inflamma-tory environment, but also work on anti-inflammatory environ-ment. This research focuses on the relations between inflamma-tory environment and MSCs autophagy's interaction and feed-back regulation, providing a train of thought for research of MSCs in inflammatory environment.

Effects of Astragalus Lily Granules on enhanced hypoxia tolerance in mice exposed to sodium nitrite poisoning / 西安交通大学学报(医学版)

Objective To study the effect of Astragali Lilium Granules on the survival time,CAT,ATP,and pathological changes in mice exposed to sodium nitrite (NaNO2)poisoning.Methods We randomly divided 60 male mice of SPF grade KM into blank group,positive control group,model group and Astragali Lilium Granules low-,medium-and high-dose groups.After 3 days of normal feeding,blank group and model group received intragastric gavage of normal saline,and positive control group was given Rhodiola rosea capsule solution.Astragali Lilium Granules groups received intragastric gavage according to the concentration (1.75,3.5 and 7 mg/kg per day) for 30 days. One hour after the last administration, all groups except the blank control group received intraperitoneal injection of NaNO2(200 mg/kg,injection amount of 0.1 mL/g).We recorded the mice's survival time,observed the pathological changes of brain,lung and heart tissues by HE staining under the microscope.The activity of CAT and the level of ATP in brain tissue were determined by colorimetric analysis.Results Compared with the blank group,the survival time in model group [(9.64±1.60)min,P＜0.05]was significantly lower;in model group,the lung tissue showed extensive pulmonary edema,red cell exudation,emphysema,and local atrophy of the lung;the interstitium of the myocardium was dilated and congested,and the local cells in the epicardium became edematous and denatured;brain cells showed necrosis,cytoplasm condensation,pyknosis,and perinuclear vacuoles.The tissues were loose and the spaces between the vessels and nerve cells widened;the activity of CAT decreased and the content of ATP decreased in lung,heart and brain tissues (P＜0.05).Compared with the model group,the survival time was significantly prolonged in Astragali Lilium Granules low-[(12.78±2.20)min]and medium-dose [(13.22±2.10)min]groups (P＜0.05).Compared with model group,the three dose groups had lessened lung tissue edema and emphysema,cardiac interstitial vascular dilatation and congestion,necrosis and nuclear pyknosis of brain nerve cells;the activity of CAT increased and the content of ATP increased in lung,heart and brain tissues (both P＜0.05).Conclusion Astragali Lilium Granules can improve the toxicity of NaNO2in mice and thus effectively protect against NaNO2-induced anoxic injury.

Influence of Guiqiyiyuan Ointment on the expression of caspase-3 and caspase-9 in rat lung tissues injured by heavy ion radiation-induced bystander effect / 解放军医学杂志

Objective To explore the influence of Guiqiyiyuan Ointment on the expressions of caspase-3 and caspase-9 in the lung and kidney of the rats damaged by heavy ion (12C6+) radiation-induced bystander effect.Methods The Wistar male rats were equally and randomly divided into seven groups,normal control group (NCG),radiation alone group (RAG) and Chinese medicine group (CMG),with the latter two groups being redivided into 6,12 and 24h groups according to the executing time.The Chinese medicine groups were given Guiqiyiyuan Ointment by gavage for two weeks in advance.The normal control group and the radiation alone groups were given the equal normal saline.Afterwards,the right lung of the rats in the radiation alone groups and Chinese medicine groups were radiated by 2Gy 12C6+ ion once.The rats in normal control group were not radiated.All groups of rats were executed 6,12,and 24h after radiation.The protein and mRNA expressions of caspase-3 and caspase-9 in the right lung,left lung and left kidney were examined with immunohistochemistry and Q-PCR.Results Compared with the normal control group,the mRNA expressions of caspase-3 and caspase-9 in the right lung,left lung and left kidney in the radiation alone groups obviously increased 6 and 24h after radiation.While the protein expressions of caspase-3 and caspase-9 in the radiation alone group obviously increased only 24h after radiation (P＜0.01).Compared with the radiation alone groups,the expressions of protein and mRNA ofcaspase-3 and caspase-9 were obviously down-regulated in the Chinese medicine groups (P＜0.01).Conclusion By controlling the up-regulation of the expression ofcaspase-3 and caspase-9,Guiqiyiyuan Ointment can prevent the lung and kidney cell apoptosis and alleviate the damage caused by heavy ion radiation-induced bystander effect in vivo.

IL-6 and TNF-α promote the differentiation of human BMSCs to TFAs by mimicry inflammatory microenvironment / 基础医学与临床

Objective To observe whether BMSCs differentiate into TAFs in inflammatory microenvironment simula-ted by IL-6 and TNF-α.Methods The experiment was divided into 9 groups:the BMSCs group,the 50 ng/mL, 100 ng/mL IL-6 intervention group, the 50 ng/mL,100 ng/mL TNF-α intervention group, the 50 ng/mL IL-6+50 ng/mL TNF-α intervention group,the 50 ng/mL IL-6+100 ng/mL TNF-α intervention group,the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group and the 100 ng/mL IL-6+100 ng/mL TNF-α intervention group; After continuous induction for 42 days, BMSCs cell morphology and cycle, TAFs-tagged α-SMA and FAP protein were examined by phase-contrast microscope,flow cytometry and Western blot method. Results Compared with the normal control group, the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group BMSC cell proliferation was significantly promoted;G1phase decreased in proportion and S phase increased;α-SMA and FAP protein expression was signifi-cantly increased (P＜0.05).Conclusions Microenvironment simulated by the 100 ng/mL IL-6+50 ng/mL TNF-α may induce the abnormal change of the BMSCs morphological and proliferative characteristics. TAFs reference mole-cule α-SMA and FAP expression is increased.

Astragalus polysaccharide attenuates liver injury in rats with chronic cadmium exposure / 基础医学与临床

Objective To explore the effects of Astragalus polysaccharides on liver injury induced by cadmium chloride. Methods The cadmium chloride solution was administrated i.v. to develope rat model of liver injury. Rats were randomized divided into three groups;control group, model group, Astragalus polysaccharides intervention group. After the 5 weeks, the rats were sacrificed and the livers were weighted, alanine aminotrans ferase(ALT), aspartate aminotransferase (AST) and lactate dehydrogenase(LDH), the contents of cadmium(Cd) was checked by plasma mass spectrometerthe(ICP-MS), the contents of interleukin-2(IL-2) and transforming growth factor-β1(TGF-β1) was measured by ELISA, the the protein expression of Bcl-2 and Bax was observed by immunohisto-chemistry staining method. Results The liver weight was increased in model group, the level of ALT, AST and LDH were also ascended in model group, the contents of Cd, TGF-β1 and the protein expression of Bax all increased in mode group(P＜0.05 or P＜0.01). The content of IL-2 and the protein expression of Bcl-2 were decreased in mode group (P＜0.05 or P＜0.01). Compared with the model group, the liver index was decreased in Astragalus polysaccharides intervention group, the levels of ALT, AST and LDH were also reduced in Astragalus poly-saccharides intervention group, the contents of Cd, TGF-β1 and the protein expression of Bax were all dropped in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01).The content of IL-2 and the protein expression of Bcl-2 increased in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01). Conclusions Astragalus polysaccharides may reduce the oxidative stress injury of rats liver cells indued by cadmium chloride, and its mechanism may be explained by regulation of Bcl- 2/Bax protein expression.

The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats / 中国应用生理学杂志

OBJECTIVES@#To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism.@*METHODS@#Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot.@*RESULTS@#Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05).@*CONCLUSIONS@#Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.

Protective effects of Astragalus immortal prescription against liver oxidative stress in irradiated mice / 解放军医学杂志

Objective To explore the protective effect and possible mechanism of Astragalus immortal prescription (AIP) from hepatic oxidative stress in mice irradiated by X ray.Methods Six to eight weeks old Kunming mice were randomly divided into normal control group,irradiation alone group,positive control group,and low,medium and high dose groups of AIP (10 rats in each group),physiological saline,thymus peptide and different concentration decocta of AIP were given respectively.After continuous gavage for 10 days,the whole body was irradiated with X ray at the dose of 8 Gy at a time,and then continued lavaging for 3 days,with weighing the mice and observing the diet,water intake and general condition.After killing the mice,the liver was take and the liver index was calculated;the morphological changes of the liver tissues were observed and the expressions of superoxide dismutase (SOD),malondiadehyde (MDA),glutathione peroxidase (GSH-Px) and nuclear factor erythroid 2-related factor 2 (Nrf2) in the liver tissues were detected.Results Compared with the normal control group,the weight of mice in model group decreased obviously (P＜0.01);pathological section showed that the liver cells were disordered,the central vein was congested and blocked seriously,the surrounding liver cells were denatured and necrotic;the activity of SOD and GSH-Px in liver tissues decreased,and the content of MDA increased (P＜0.05),the expression of Nrf2 protein increased significantly (P＜0.01).Compared with the radiation alone group,the weight of mice in positive control group and high dose AIP group increased significantly (P＜0.01);pathological section showed that the liver tissue structure was basically normal,the liver cells were radially arranged with the central vein as the center;the activity of SOD and GSH-Px in liver tissues increased,and the content of MDA decreased (P＜0.05),the expression of Nrf2 protein decreased significantly (P＜0.01).Conclusion AIP has protective effect from hepatic oxidative stress in mice irradiated by X ray,and the mechanism may be related to its antioxidant effects.

Protective effects of Angelica sinensis and Angelica sinensis polysaccharides against liver and kidney injury induced by irradiation in rats / 解放军医学杂志

Objective To investigate the protective effects of Angelica sinensis and Angelica sinensis polysaccharide against liver and kidney injury induced by irradiation of SD rats.Methods Thirty-two SD rats were randomly divided into four groups:control group,model group,Angelica decoction group and Angelica sinensis polysaccharide group.After 14 days of routine feeding,the rats were treated by intragastric administration once a day for 7 days.From the 8th day,the rats received whole body X-ray irradiation for 2 days.The total absorbed dose was 6Gy,and the dose rate was 92.26cGy/min.The rats were killed and the blood were collected from the femoral artery 3 days after irradiation.The content of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and lipid peroxide (LPO) in the liver and kidney tissues was detected by colorimetric analysis.The pathological changes of the liver and kidney were observed by HE staining under microscope.The expression of Nrf2 protein in the liver and kidney of the rats was detected by Western blotting.Results Compared with control group,the weight of the liver and kidney significantly increased,the activities of SOD and GSH-Px significantly decreased,the contents of MDA and LPO significantly increased,and the liver and kidney cells showed obvious edema,and the content of Nrf2 in the liver and kidney increased in the model group.Compared with model group,the weight of the liver and kidney decreased,SOD and GSH-Px activities significantly increased,the contents of MDA and LPO significantly decreased,edema of the liver and kidney significantly reduced,the expression of Nrf2 in the liver and kidney tissues decreased in Angelica decoction group and Angelica sinensis polysaccharides group,while there was no difference between Angelica sinensis polysaccharide group and Angelica decoction group.Conclusion Angelica sinensis and Angelica polysaccharides have a good protective effect against liver and kidney injury induced by radiation in SD rats,which is implemented mainly through influencing the expression of free radicals.The protective effect of Angelica against radiation injury is achieved mainly by Angelica polysaccharide.