Objective To explore the mechanism of Wenyang Shengji Ointment(温阳生肌膏,WYSJO)in the treatment of diabetic wounds from the perspective of network pharmacology,and to veri-fy it by animal experiments. Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and related literature were used to screen active compounds in WYSJO and their corresponding targets.GeneCards,Online Mendelian Inheritance in Man(OMIM),DrugBank,PharmGkb,and Therapeutic Target Database(TTD)databases were employed to identify the targets associated with diabetic wounds.Cytoscape 3.9.0 was used to map the ac-tive ingredients in WYSJO,which was the diabetic wound target network.Search Tool for the Retrieval of Interaction Gene/Proteins(STRING)platform was utilized to construct protein-protein interaction(PPI)network.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analyses were performed to identify signaling pathways be-tween WYSJO and diabetic wounds.AutoDock 1.5.6 was used for molecular docking of core components in WYSJO to their targets.Eighteen rats were randomly divided into control,model,and WYSJO groups(n=6).The model and WYSJO groups were used to prepare the model of refractory wounds in diabetes rats.The wound healing was observed on day 0,5,9,and 14 after treatment,and the wound tissue morphology was observed by hematoxylin-eosin(HE)staining.The expression levels of core genes were detected by quantitative real-time polymerase chain reaction(qPCR). Results A total of 76 active compounds in WYSJO,206 WYSJO drug targets,3 797 diabetic wound targets,and 167 diabetic wound associated WYSJO targets were screened out through network pharmacology.With the use of WYSJO-diabetic wound target network,core targets of seven active compounds encompassing quercetin,daidzein,kaempferol,rhamnetin,rham-nocitrin,strictosamide,and diisobutyl phthalate(DIBP)in WYSJO were found.GO enrich-ment analysis showed that the treatment of diabetes wounds with WYSJO may involve lipopolysaccharide,bacteria-derived molecules,metal ions,foreign stimuli,chemical stress,nutrient level,hypoxia,and oxidative stress in the biological processes.KEGG enrichment analysis showed that the treatment of diabetes wounds with WYSJO may involve advanced glycation end products(AGE-RAGE),p53,interleukin(IL)-17,tumor necrosis factor(TNF),hypoxia inducible factor-1(HIF-1),apoptosis,lipid,atherosclerosis,etc.The results of animal experiments showed that WYSJO could significantly accelerate the healing process of diabetic wounds(P<0.05),alleviate inflammatory response,promote the growth of granulation tis-sues,and down-regulate the expression levels of eight core genes[histone crotonyltrans-ferase p300(EP300),protoc gene-oncogene c-Jun(JUN),myelocytomatosis(MYC),hypoxia inducible factor 1A(HIF1A),mitogen-activated protein kinase 14(MAPK14),specificity pro-tein 1(SP1),tumor protein p53(TP53),and estrogen receptor 1(ESR1)]predicted by the net-work pharmacology(P<0.05). Conclusion The mechanism of WYSJO in treating diabetes wounds may be closely related to AGE-RAGE,p53,HIF-1,and other pathways.This study can provide new ideas for the phar-macological research of WYSJO,and provide a basis for its further transformation and appli-cation.

Objective To investigate the effects of curcumin on acute kidney injury in endotoxic shock rabbits and discuss the possible mechanism.Methods Forty healthy male New Zealand white rabbits aged 2 months were randomly divided into 4 groups:control group (group C),curcumin control group (group Cur,curcumin 50 mg/kg),endotoxic shock group (group L,LPS 5 mg/ kg) and curcumin pretreatment group (group CurL,curcumin 50 mg/kg,LPS 5 mg/kg 30 minutes later),10 in each group.The concentrations of blood urea nitrogen (BUN) and creatinine (Cr) were detected,and the rabbits were sacrificed 6 h after LPS treatment.The pathological changes of renal tissue were examined and the pathological scores were recorded.SOD activities were measured by xanthine oxidase method.MDA contents were assayed according to thiobarbituric acid method.Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to investigate the mRNA levels of Nrf2 and HO 1.The expressions of Nrf2 total protein,Nrf2 nuclear protein and HO-1 protein in the renal tissue were determined by Western blot.Results No abnormal structures were visible in groups C and Cur.The histopathological changes of the kidneys included glomerular shrinkage,cellu lar swelling,vacuolization and desquamation in tubules,interstitial edema and massive inflammatory cells infiltration in group L.Compared with group C,the pathological manifestations was relieved obviously in group CurL.Compared with group C,the pathological score,the concentrations of BUN,Cr,and MDA were obviously increased,while SOD activity was significantly decreased,the mRNA expression of Nrf2 and HO-1,the protein levels of Nrf2 total,Nrf2 nuclear and HO-1 in the renal tissues were distinctly increased in groups L and CurL (P＜0.05).Compared with group L,the pathological score,the concentrations of BUN,Cr and MDA were obviously decreased,while SOD activity was significantly increased,the mRNA expression of Nrf2 and HO 1,the protein levels of Nrf2 total,Nrf2 nuclear and HO-1 in the renal tissues were significantly increased in group CurL (P ＜0.05).Conclusion Curcumin can ameliorate acute kidney injury induced by endotoxic shock in rabbits,and its mechanism may be relate to activation of Nrf2/HO-1 signaling pathway.

Objective To observe the effect of erythropoietin(EPO)in rats with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP). Methods A total of 90 male adult SD rats were randomly divided into three groups:blank control group(BC group),air control group(AC group)and DEACMP group. Time points(1,3,7,14,21 and 28 days after acute carbon monoxide poisoning)were set for measuring the changes. Pure CO were injected into the abdominal cavity of the rats from DEACMP group for several times to estab-lish DEACMP model. Rats in AC group were injected with equal volume of air by the same way. BC group were without any treatment. Morris water maze test was used to measure the cognitive behavior. The apoptosis of pyrami-dal neurons at hippocampus was measured by TUNEL. The expression of erythropoietin(EPO)in hippocampus was measured by immunohistochemistry. Results The average escaped latency of rats in DEACMP group increased after poisoning compared with rats in other two groups(P<0.05). The apoptosis of pyramidal neurons in hippocam-pus increased from day 1 after the CO poisoning. It reached the peak at day 7 and it still had a high expression at day 28. The apoptotic index in DEACMP group increased significantly compare with that of BC group and AC group (P < 0.05). The expression of EPO in hippocampus was found increased from day 1 after the CO poisoning and reached the peak at day 3. It began to reduce at day 7. The expression in DEACMP group was higher than those of other two groups (P < 0.05). Conclusions It may be one of the causes of the DEACMP that the expression of EPO decreased in the middle and late stage after CO poisoning and its anti-apoptosis was decreased.

Objective To observe the effect of excretory/secretory products from Trichinella spiralis adult worms(AES)on cecal ligation and puncture(CLP)?induced sepsis in mice. Methods Forty?eight BALB/c mice were randomly divided into 3 groups:a sham operation group(PBS+sham group,Group A),a CLP?induced sepsis group(PBS+CLP group,Group B)and an AES treatment group(AES+ CLP group,Group C). The mice of each group were intraperitoneally injected with 25 μg of AES or PBS only as a control in a total volume of 200μl. Eight mice from each group were selected randomly for survival analy?sis of 96 hours. The other 8 mice in each group were observed for pathological changes in the lung,liver and kidney tissues by HE staining 12 h after CLP,and then determined for the detection of cytokines including TNF?α,IL?1β,IL?6,IL?10 and TGF? βin the sera by ELISA. Results The difference among the survival rates of mice in the 3 groups was statistically significant (χ2=21.16,P<0.05). Compared to Group A(100%),the survival rate of mice in Group B(0)decreased significantly(P<0.05),and also the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group B increased signifi?cantly after CLP. Compared with the mice in group B,the survival rate of those in Group C(70%)increased significantly(P<0.05),and the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group C decreased significantly after the treatment with AES. The differences among the levels of pro?inflammatory cytokines TNF?α(F=27.11,P<0.05),IL?1β(F=18.75,P<0.05)and IL?6(F=100.93,P<0.05)in the sera of the mice in the three groups were statistically signifi?cant. Compared with the mice in Group A,the levels of the 3 cytokines of those in Group B increased significantly(all P <0.05). However,after the treatment with AES,the levels of the pro?inflammatory cytokines of those in Group C decreased signifi?cantly(all P<0.05). The differences among the levels of immunoregulatory cytokines IL?10(F=10.88,P<0.05)and TGF?β(F=11.37,P<0.05)in the sera of the mice in the three groups were also statistically significant. Compared with the mice in Group B,the levels of IL?10 and TGF?β of those in Group C were higher after treatment with AES(both P<0.05). Conclu?sion T. spiralis AES has a therapeutic potential for alleviating sepsis induced by CLP in mice.

Objective To investigate the relationship between the C677T polymor-phism of the methylenetetrahydrofolate reductase (MTHFR) gene/G448A polymorphism of the