OBJECTIVE To explore the molecular mechanism of Xijiao Dihuang Decoction(XJDHT)in inhibiting inflammatory response in sepsis based on network pharmacology,molecular docking and in vitro and in vivo experiments.METHODS Active com-ponents of XJDHT were screened using the TCMSP and HERB databases.Sepsis-related targets and histone H3K36 trimethylation(H3K36me3)-associated targets were retrieved from GeneCard,OMIM,and DisGeNet databases.A protein-protein interaction(PPI)network was constructed using the STRING database,and core targets were identified via Cytoscape 3.9.1.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to predict potential mechanisms.Mo-lecular docking validated ligand-receptor binding affinity.A cecal ligation and puncture(CLP)model was established in mice to evalu-ate 24-hour sepsis scores(MSS)and survival rates.Blood routine parameters were analyzed using an automated hematology analyzer.Serum IL-1β and TNF-α levels were measured by ELISA.Liver histopathology was assessed via HE staining,and H3K36me3 expression in Kupffer cells was detected by immunofluorescence.In vitro,LPS-induced THP-1 cells were used as an in-flammatory model.ChIP-qPCR evaluated H3K36me3 enrichment at IL-1β and TNF-α gene loci.Western blot analyzed HIF-1α,EGFR,and AKT1 pathway proteins.RESULTS A total of 28 active components of XJDHT were identified,corresponding to 987 gene targets,with 189 overlapping sepsis-related targets.Core targets included TNF,IL1B,and GAPDH.Enriched pathways includ-ed EGFR tyrosine kinase inhibitor resistance.Molecular docking confirmed strong binding between core components and key targets.In vivo,compared to the sham group,the CLP group exhibited significantly reduced 24-hour survival(P<0.01),elevated MSS(P<0.01),immune imbalance,and increased serum IL-1β and TNF-α levels(P<0.01).High-and low-dose XJDHT intervention im-proved survival(P<0.01),reduced MSS(P<0.01),restored immune homeostasis,and dose-dependently suppressed IL-1β and TNF-α(P<0.01).CLP mice showed elevated H3K36me3 in Kupffer cells and severe hepatic inflammation,while XJDHT dose-de-pendently reduced H3K36me3(P<0.05)and attenuated liver injury.In peritoneal macrophages,CLP upregulated H3K36me3,IL-1β,TNF-α,HIF-1α,p-AKT1/AKT1,and EGFR(P<0.01),which were reversed by XJDHT(P<0.05,P<0.01).In vitro,LPS increased H3K36me3 and IL-1β and TNF-α expression(P<0.01),with ChIP-qPCR confirming H3K36me3 enrichment at IL-1β lo-ci(P<0.01).Treatment with 15%XJDHT-containing serum for 24 h reduced H3K36me3(P<0.01),diminished its recruitment to IL-1β loci(P<0.01),and inhibited LPS-induced activation of EGFR,HIF-1α,and p-AKT1/AKT1(P<0.05,P<0.01).HIF-1α agonist Dimethyloxallyl Glycine(DMOG)further validated that XJDHT suppressed H3K36me3-mediated epigenetic remodeling via HIF-1α-related pathways.CONCLUSION XJDHT inhibits inflammatory responses,regulates immune homeostasis,and improves sepsis prognosis,potentially by modulating H3K36me3 epigenetic modifications at IL-1β loci.

OBJECTIVE To explore the molecular mechanism of Xijiao Dihuang Decoction(XJDHT)in inhibiting inflammatory response in sepsis based on network pharmacology,molecular docking and in vitro and in vivo experiments.METHODS Active com-ponents of XJDHT were screened using the TCMSP and HERB databases.Sepsis-related targets and histone H3K36 trimethylation(H3K36me3)-associated targets were retrieved from GeneCard,OMIM,and DisGeNet databases.A protein-protein interaction(PPI)network was constructed using the STRING database,and core targets were identified via Cytoscape 3.9.1.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to predict potential mechanisms.Mo-lecular docking validated ligand-receptor binding affinity.A cecal ligation and puncture(CLP)model was established in mice to evalu-ate 24-hour sepsis scores(MSS)and survival rates.Blood routine parameters were analyzed using an automated hematology analyzer.Serum IL-1β and TNF-α levels were measured by ELISA.Liver histopathology was assessed via HE staining,and H3K36me3 expression in Kupffer cells was detected by immunofluorescence.In vitro,LPS-induced THP-1 cells were used as an in-flammatory model.ChIP-qPCR evaluated H3K36me3 enrichment at IL-1β and TNF-α gene loci.Western blot analyzed HIF-1α,EGFR,and AKT1 pathway proteins.RESULTS A total of 28 active components of XJDHT were identified,corresponding to 987 gene targets,with 189 overlapping sepsis-related targets.Core targets included TNF,IL1B,and GAPDH.Enriched pathways includ-ed EGFR tyrosine kinase inhibitor resistance.Molecular docking confirmed strong binding between core components and key targets.In vivo,compared to the sham group,the CLP group exhibited significantly reduced 24-hour survival(P<0.01),elevated MSS(P<0.01),immune imbalance,and increased serum IL-1β and TNF-α levels(P<0.01).High-and low-dose XJDHT intervention im-proved survival(P<0.01),reduced MSS(P<0.01),restored immune homeostasis,and dose-dependently suppressed IL-1β and TNF-α(P<0.01).CLP mice showed elevated H3K36me3 in Kupffer cells and severe hepatic inflammation,while XJDHT dose-de-pendently reduced H3K36me3(P<0.05)and attenuated liver injury.In peritoneal macrophages,CLP upregulated H3K36me3,IL-1β,TNF-α,HIF-1α,p-AKT1/AKT1,and EGFR(P<0.01),which were reversed by XJDHT(P<0.05,P<0.01).In vitro,LPS increased H3K36me3 and IL-1β and TNF-α expression(P<0.01),with ChIP-qPCR confirming H3K36me3 enrichment at IL-1β lo-ci(P<0.01).Treatment with 15%XJDHT-containing serum for 24 h reduced H3K36me3(P<0.01),diminished its recruitment to IL-1β loci(P<0.01),and inhibited LPS-induced activation of EGFR,HIF-1α,and p-AKT1/AKT1(P<0.05,P<0.01).HIF-1α agonist Dimethyloxallyl Glycine(DMOG)further validated that XJDHT suppressed H3K36me3-mediated epigenetic remodeling via HIF-1α-related pathways.CONCLUSION XJDHT inhibits inflammatory responses,regulates immune homeostasis,and improves sepsis prognosis,potentially by modulating H3K36me3 epigenetic modifications at IL-1β loci.

Objective To Analyze the expression profiles of LncRNAs and mRNAs in ovarian epithelial cancer cell lines by gene mi-croarray, and then provide experimental evidences for investigating the function of LncRNAs associated with ovarian cancer. Methods The differentially expressed LncRNAs and mRNAs in ovarian epithelial cancer cell lines, such as A2780, HO8910 and SKOV3, and ovarian epithelial cell line HOSEpiC were analyzed by gene microarray. The differentially expressed mRNAs were further performed the KEGG pathway enrichment analysis. The expression levels of six candidate LncRNAs, which had significant difference between the o-varian epithelial cancer cell line and the ovarian epithelial cell line, were further verified by qRT-PCR. Results There were 227 up-regulated LncRNAs and 483 down-regulated LncRNAs in A2780, HO8910 and SKOV3 cell lines. The differentially expressed mRNAs in A2780, HO8910 and SKOV3 cell lines were mainly enriched in the tumor-related pathways such as PI3K-AKT, mTOR and TNF-α( P<0.05) . The expression levels of PTPRG-AS1, CCNT2-AS1, XLOC 009869 and LINC01138 in ovarian epithelial cancer A2780, SKOV3 and OVCR3 cell lines were up-regulated (P<0.05), while those of RP11-252P19.2 and RP11-744I24.2 in ovarian epithelial cancer A2780, SKOV3, OVCR3 and 3AO cell lines were down-regulated ( P<0.05) . Conclusion The differentially expressed LncR-NAs and mRNAs in ovarian epithelial cancer cell lines may be obtained by gene microarray, and the differentially expressed mRNAs are associated with the tumor-related pathways such as PI3K-AKT, mTOR and TNF-α, which may provide new targets for the diagnosis and treatment of ovarian cancer.

Objective To investigate the effect of early combined functional exercise on lower limb rehabilitation of vessel bridge taken in coronary artery bypass surgery. Methods From January to October 2017,90 patients undergoing coronary artery bypass surgery were randomly divided into control group and experimental group by coin toss, with 45 patients in each group. The control group was given routine nursing care, and the experimental group was given early combined functional exercise (foot dorsiflexion, ring rotation exercise and straight leg raising exercise) on the basis of routine nursing care. The difference of correlation index between two groups was compared. Results In the experimental group, the number of swelling in the thigh, crus and ankle were 20, 25 and 35 respectively; the control group was 32, 35, 43, the difference was statistically significant(χ2=6.55, 5.00, 8.38, P<0.05). The remission of limb swelling in the experimental group was also better than that in the control group(P<0.01). The incidence of limb wound complications: 6 cases in the experimental group, 18 cases in the control group(χ2=11.79,P<0.01));limb muscle strength grade Lovett:experimental group 0 case with grade 3,19 cases with grade 4,26 cases with grade 5,control group 9 cases with grade 3,26 cases with grade 4,10 cases with grade 5, there were significant differences between two groups (χ2=17.20, P<0.05). The limb comfort score,postoperative bed time,postoperative hospital stay was(7.11±1.15)points,(5.28±1.07)d, (11.04±2.01)d in the experimental group and(5.18±1.13)points,(6.97±1.21)d,(12.67±1.59)d in the control group,the differences were significant(t=-8.02,6.97,4.23,P<0.01). Conclusions To carry out early functional exercise can not only take angiogenesis effectively reduce limb swelling rate after coronary artery bypass grafting,relieve the severity of limb swelling,but also can reduce the incidence of incision complications,promote the recovery of limb muscle strength,shorten the postoperative recovery time and postoperative hospitalization time,is worthy of promotion and use in clinical nursing work.

Objective To explore the expression of transient receptor potential channel 6(TRPC6)in human breast cancer cells, and to clarify the correlation of TRPC6 with the invasion potential of breast cancer cells. Methods The human breast cancer cell strains MCF-7 (hypo-invasion group)and MDA-MB-231 (hyper-invasion group)were cultured.The expressions of TRPC6 mRNA and protein in in two groups were detected by RT-PCR and Western blotting methods.Then the MDA-MB-231 cells were divided into control group and SKF96365 group, the effects of SKF96365 on the invasion ability of MDA-MB-231 cells invitro were explored by wound healing assay and Transwell experiment.Results The results of Western blotting and RT-PCR showed that the expression levels of TRPC6 mRNA and protein in MDA-MB-231 cells were higher than that in MCF-7 cells(P<0.05).The wound healing assay showed the numbers of migrating cells in 5,25 and 40μmol·L-1 SKF96365 groups (76.24±7.54, 45.33±4.50,25.12±1.57)were lower than those in control group (130.48±9.55)(P<0.05).The Transwell experiment results indicated that the invasiveness of MDA-MB-231 cells were inhibited significantly by SKF96365 compared with control group (P<0.05).Conclusion The invasion ability of human breast cancer MDA-MB-231 cells is promoted by upregulating the TRPC6 expression, which indicates that the TRPC6 may play role in the metastasis of human breast cancer.

Objective To investigate the minimal and optimal concentration of ropivacain for epidural anesthesia in patients with breast cancer.Methods 103 patients with breast cancer ASA Ⅰ-Ⅱ undergoing elective radical mastectomy were conducted with epidural anesthesia.T2-3 were selected as a puncture site.A catheter was inserted into the epidural space in a cephalic direction for 3.5-4.0 cm.The concentration of ropivacain was produced by the test of up-down sequential allocation technique.20-25 ml of 0.20 ％ ropivacaine was given after a test of 0.20 ％ 5 ml ropivacaine.If the anesthesia was effective (VAS≤3),the next concentration was down 0.01％,while the VAS was more than 5,the next concentration was up 0.01％.The anesthesia effect and segments of block were measured with acupuncture.Blood pressure (NIBP),heart rates(HR),blood oxygen saturation (SPO2),blood gas and VAS were monitored during operation.Nasal catheter oxygen inhalation was used in routine method.Results The median effective concentration (EC50) of ropivacaine was 0.10 ％ determined by the formula of dixon and massey.The 95 ％ confidence intervals was 0.1022 ％-0.1065 ％.The least and the best analgesic concentration of ropivacaine for the upper thoracic epidural block was 0.14 ％ and 0.17 ％-0.18 ％ respectively.Conclusion The minimal and the optimal analgesic concentration of ropivacaine for the upper thoracic epidural block are 0.14 ％ and 0.17 ％-0.18 ％respectively,and which provides a safe and rational use of ropivacaine for the clinic.

Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.

Three types of microscopic operations were used in this study: 1. excision of the optic vesicle alone; 2. after excising the optic vesicle, a piece of forebrain tissue taken from another donor was inserted in between presumptive lens ectoderm and forebrain wall; 3. cut down the optic vesicle and portion of forebrain tissue and replaced them back in situ by turning over 180?. The operations were carried out on Rana nigromaculata at 15 and 16 stages, in order to bring the presumptive lens ectoderm to come into contact directly with the forebrain tissue for the purpose to analyse the possibility of eye cup formation from the brain wall. The results indicate that the forebrain wall which came into contact with the presumptive lens ectoderm could be induced and differentiated into a secondary eye cup or retina. A total of 81 cases of secondary eye cups among 282 operations (28.8%) were observed. The mechanism for the induction and its significance were discussed.

The mesenchymal cells were carefully scraped off from the optic vesicles or eye cups at 15 to 24 stages in Rana limnocharis by a hair loop (those specimens from 21 to 24 stages were treated with trypsin in order to separate the mesenchymal cells easier) and then the optic vesicles or eye cups were transplanted into the coelom of tadpoles for 1 to 2 weeks. The transplantation of the eye cups Without scraping mesenchymal cells off at 21 to 24 stages were used as control. The results are as follows: 1.The pigment epithelium of eye, which was transplanted at 15 to 20 stages, (the pigment epithelium have not differentiated at those stages) can not fully develop or only a tiny piece of pigment epithelium layer occurs. 2.The pigment epithelium of eye, which was transplanted at 21 to 24 stages, (the pigment epithelium have differentiated at those stages) can develop, but separates itself from the normal position and hangs outside the eye or curls inside the cavity of eye in a vesicular form. 3.In the intact mesenchymal cells, the pigment epithelium of eye, which was transplanted at 21 to 24 stages, can develop well and remain in normal position. These results suggest that the mesenchymal cells might have an important effect on the development and morphology of eye pigment epithelium before and after its differentiation. Before differentiation of pigment epithelium, the existence of mesenchymal cells is a necessary condition for its normal differentiation; after its differentiation, the mesenchymal cells might play a mechanical role to keep the pigment epithelium layer in normal position.

This study was an attempt to excise or transplant optic vesicles, and excise or transplant presumptive lens ectoderms, by means of microscopic manipulations. Its purpose was to explore the dependence of the development of lens on the eye cup and the ability of induction of the eye cup to develop lens in liana nigromaculata, R. limnocharis, and Kaloula barealis, which are frequently used in experimental embryology in China. The results show that their lenses cannot develop without eye vesicles. This indicates that they belong to the pattern of dependent differentiation. The results also indicate that the situation of forming lenses from ectoderms under induction of eye cup is different in three kinds of frogs: the lenses can be frequently formed from ectoderm of both Lead and abdomen in Rana nigromaculata; but the frequency of lens formation from ectoderm of abdomen is rather low in R. limrocharis; and only the ectoderm of head is positive in Kaloula barealis. The interrelation in size between the eye cup and the lens was recorded. The formation of a secondary eye cup from brain wall under condition of the contact with the presumptive lens ectoderm was reported too. In addition, it was found that the temperature might have influence on eye differentiation.