Objective:To study the effects of the Gekko gecko ethanol extract on granular cells apoptosis of rat ovaries.Methods :3 month and 6 month female Wistar rats were taken in this study, and 20 rats were in each age group.Each age group were randomly divided into two groups(experimental group and control tragastric administration in experimental group, corresponding volume of 0.9% sodium chloride was given in control group by the same way.Apoptosis of granular cells and distribution of cell cycle in rat ovary were measured by terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL) and propidi-um iodide(PI) staining, AnnexinV/PI double labelling staining by flow cytometry (FCM) after administration for 15 days.Re8utts:The apoptosis numbers of granular cells in 3 month and 6 month age experimental group were 34 ±7 and 53 ±11, in control group were 48±9 and 76±17 respectively by TUNEL.There was signrficantly statistic difference (P <0.01).The apoptosis rates in 3 month and 6 month age experimental group were(6.06 ±2.01)% and (12.16 ±3.26) % , in the control group were (8.23±2.13) % and (23.69 ±6.28)% respectively by PI staining.There was significantly statistic drfference (P <0.05).However, no significant difference was found between two groups in cellular cycle distribution (P > 0.05).Early apoptosis rates in 3 month and 6 month age experimental group were (6.71±3.11) % and (23.74±6.28) % , in the control group were (19.05 ±5.78) % and (36.55± 8.35) % respectively by AnnexinV/PI double labelling staining FCM.There was significantly statistic difference (P <0.01).The apoptosis of granular cells were sig-nificantly less in experimental groups comparing wrth control groups both in 3 month and 6 month age group (P<0.01 or P<0.05).Conclusions :The ethanol extract of gekko gecko can inhibit the apoptosis of granu-lar cells in rat ovary, then improves the ovary function and delay ageing of rat.

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Objective To evaluate the effect of hyperlipidemia on pulmonary uptake in the patients inhaling sevoflurane for anesthesia.Methods A total of 103 patients of both sexes,aged 20-50 yr,with body mass index of 18-25 kg/m2,of American.Society of Anesthesiologists physical status Ⅰ or Ⅱ,undergoing elective operation under general anesthesia,were enrolled in the study.Anesthesia was induced with iv midazolam 0.1 mg/kg,vecuronium 0.1 mg/kg,fentanyl 0.l mg/kg and etomidate 0.3 mg/kg.Patients were mechanically ventilated after endotracheal intubation.Patients inhaled 2％ sevoflurane for 30 min at an oxygen flow rate of 2 L/min.The inspired concentrations (Fi) and expired concentrations (Fe,Fe≈alveolar concentration [Fa]) of sevoflurane were recorded at 1,3,5,7,10,15,20 and 30 min of inhalation (T0-7).Patients were divided into either control group group C) or hyperlipidemia group (group H) according to the results of blood lipid levels after the end of observation.The ratio of Fa/Fi and time spent in reaching the titration value (Fa/Fi =0.8) were calculated in each group.Results There were 67 cases in group C and 36 cases in group H.Compared with group C,the Fa/Fi ratio was significantly decreased at 5,7 and 10 min of inhalation,and the time spent in reaching the titration value was prolonged in group H (P ＜ 0.05).Conclusion The pulmonary uptake of sevoflurane is increased,which may be associated with increased blood/gas partition coefficient.

OBJEVTIVETo investigate the expression of transient receptor potential lvanilloidreceptor 4 (TRPV4) protein in pemphigus vulgaris (PV), bullous pemphigoid (BP), dermatitis herpetiformis (DH), and epidermolysis bullosa acquisita (EBA), and explore the role of TRPV4 in the pathogenesis of these diseases.

METHODSTRPV4 protein in normal skin tissues and lesions of PV, BP, DH, and EBA were detected with immunohistochemistry.

RESULTSThe positivity rate of TRPV4 protein expression was 61.90% in PV, 81.81% in BP, 72.22% in DH, and 68.42% in EBA. TRPV4-positive rates in these lesions were significantly lower than the rate in normal skin tissues (93.33%) and also differed significantly among these lesions (PV

CONCLUSINLow TRPV4 expressions may affect the formation and reconstitution of skin connection. TRPV4 may play an role in the occurrence and development of autoimmune bullous skin disorders.

Dermatitis Herpetiformis ; metabolism ; Diagnosis, Differential ; Epidermolysis Bullosa Acquisita ; metabolism ; Humans ; Pemphigoid, Bullous ; metabolism ; Pemphigus ; metabolism ; Skin ; pathology ; TRPV Cation Channels ; metabolism

Objective To examine whether Tim-3 plays a protective role in paraquat poisoning induced excessive immune response and tissue damage based on the critical roles of Tim-3 controlling inflammatory response.Methods A paraquat poisoning model was established in wild type and in Tim-3 transgenic C57BL/6 mice by intraperitoneal injection of paraquat (40 mg/kg) .In addition, C57BL/6 mice with paraquat poisoning were injected with Tim-3 soluble protein( sTim-3) or control protein to see the effect of Tim-3 blocking on the progression of paraquat poisoning.Samples were collected at 6 and 24 h after paraquat injection respectively and were examined for tissue damage, cytokine expression and paraquat metabolism.Results After paraquat poisoning, there was significantly attenuated tissue damage in the lungs and kidneys and decreased TNF-α,IL-6 and IL-1 beta expression in the PBMCs or in the serum from Tim-3 transgenic mice compared to wild type mice.The serum concentration of paraquat in Tim-3 transgenic mice was also significantly decreased.However, in sTim-3 treated paraquat poisoning mice, there was significantly increased cytokine expression and tissue damage compared to control protein treated mice.The in vitro data showed that Tim-3 signaling negatively regulated macrophages mediated inflammatory response.Conclusion Tim-3 plays a critical role in maintaining the homeostasis after paraquat poisoning. Further investigation on the regulatory roles of Tim-3 in inflammation will shed new light on the pathogenesis of paraquat poisoning and provide new therapeutic strategies.

Objective To develop a human Tim-3 specific monoclonal antibody and evaluate its biological activity and possible use in clinical diseases associated with dysregulated Tim-3 expression .Methods The BALB/c mice were immu-nized by conventional method, and positive clones were used to develop anti-human Tim-3 antibody, the binding and neutralization activities of which in vitro and in vivo were investigated.Results ①A monoclonal antibody (clone L3D) which could specifically bind to human Tim-3 protein in ELISA assay was obtained and the subtype of the monoclonal antibody was IgG2a .②Flow cytometry indicated that the monoclonal antibody could bind to Tim-3 expressed in human U937 cells.This antibody also showed a cross activity to mice′Tim-3.③The monoclonal antibody inhibited the apoptosis of THP1 cells induced by Gal-9, the ligand of Tim-3.④Injection of Tim-3 antibody exacerbated sepsis in mice as marked by the decreased survival rate and increased expression of pro-inflammatory cytokines .Conclusion An anti-human Tim-3 monoclonal antibody is successfully obtained.The excellent binding and neutralization activities of this antibody enable it to be widely used in clinical diseases associated with deregulated Tim-3 expression .

Objective Based on the gene expression omnibus(GEO)database,bioinformatics methods were employed to analyze the expression characteristics of hypoxia-related differentially expressed genes(HRDEGs)in ischemic stroke,and key genes were screened,to provide important support for a deeper understanding of ischemic stroke.Methods The GSE16561 and GSE58294 datasets were downloaded from the GEO database,and Python software was used for data integration.The Combat method was employed to eliminate batch effects while retaining disease grouping characteristics.Principal component analysis was conducted to reduce dimensionality of the data before and after batch effect removal,and intraclass correlation coefficient(ICC)testing was performed on the ischemic stroke and normal control groups.Gene set enrichment analysis(GSEA)and single-sample GSEA were conducted on the merged and batch effects eliminated dataset,with a nominal P-value(NOM P-val)＜0.05 and false discovery rate P-value(FDR P-val)＜0.25 used as criteria to select significantly different gene sets.Differential expression genes between the ischemic stroke samples and normal control samples after merging and eliminating batch effects of the GSE16561 and GSE58294 datasets were identified using R software,with an absolute value of log2 gene expression fold change(FC)≥0.58 and adjusted P-value(Padj)＜0.05 as selection criteria.Intersection with hypoxia-related genes obtained from the National Center for Biotechnology Information(NCBI)in the United States yielded the HRDEGs.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed on the HRDEGs,and the STRING database was used to construct a protein-protein interaction network of differentially expressed genes.The top 10 key genes were filtered using Cytoscape 3.8 software.Results The ICC analysis results showed excellent consistency in the ischemic stroke and normal control samples after batch effect removal,with ICC values of 0.94 and 0.98 for the GSE16561 and GSE58294datasets,respectively.GSEA results demonstrated significant enrichment of 34 gene sets in the stroke samples in the newly merged and batch effects removed dataset from GSE16561 and GSE58294,leading to the identification of 404 differentially expressed genes(all with Padj＜0.05),including 354 upregulated genes and 50 downregulated genes.Intersection with hypoxia-related genes yielded 64 HRDEGs.GO enrichment analysis indicated significant enrichment of HRDEGs in vesicle lumen,cytoplasmic vesicle lumen,secretory granule lumen,with molecular functions such as amide binding,peptide binding,phospholipid binding,and enzyme inhibitor activity.These genes are primarily involved in the positive regulation of cytokine production,regulation of immune response,response to bacterium-derived molecules,and response to lipopolysaccharide,among other biological processes.KEGG enrichment analysis revealed enrichment of HRDEGs in pathways related to lipid and atherosclerosis,Salmonella infection,neutrophil extracellular trap formation,nucleotide-binding oligomerization domain-like receptor signaling pathway,protein glycosylation in cancer,tuberculosis,and necroptosis.Based on the protein-protein interaction network,10 key genes were identified,including arginase1(ARG1),caspase1(CASP1),interleukin1 receptor type 1(IL-1R1),integrin subunit alpha M(ITGAM),matrix metalloproteinase9(MMP9),prostaglandin-endoperoxide synthase 2(PTGS2),signal transducer and activator of transcription 3(STAT3),Toll-like receptor2(TLR2),TLR4,and TLR8.Conclusion This study has identified 10 key genes associated with ischemic stroke and hypoxia through bioinformatics mining,which maybe provid potential targets for subsequent research and diagnostic and therapeutic interventions.