Objective To observe the effects of vitamin K3(VK3) on cell growth and apoptosis in the cell line SMMC-7721 of human liver cancer and the changes in expression of survivin gene.Methods The inhibitory effect of VK3 on SMMC-7721 cells was examined by CCK-8 (cell counting kit).Morphological evaluation of apoptosis was performed by Hoechst33342 staining.The amount of apoptotic cell was assayed by flowcytometry.The changes of survivin gene expression were detected by RT-PCR.Results The inhibitory rate of VK3 (at concentrations of 2,5,10,20,25 mmol/L for 48 h) on SMMC-7721 growth was 3.8%,50.1%,63.9%,78.5%,84.7% respectively.The cells in apoptosis were strongly stained by Hoechst33342 compared with the cells in control group.The apoptostic rate of cells was 33.28%,63.97%,77.69%,87.30% and 92.40%.Following exposure to VK3 at the concentration of 2,5,10,20 and 25 mmol/L for 48h,the survivin mRNA expression in SMMC-7721 was downregulated.Conclusion VK3 inhibits the proliferation of SMMC-7721 cells and induce apoptosis.The mechanism may related to survivn gene.

Objective:To explore the effects and mechanism of liraglutide on nitrogen monoxide (NO) release in human umbilical vein endothelial cells in the state of hypoxia and high glucose.Methods:A model of hypoxia and high glucose was established by using isolation and culture of primary human umbilical vein endothelial cells (HUVECs) in vitro.HUVECs were incubated with liraglutide and/or exendin (9-39) for 4 h.The metabolic ability of cells was detected by MTT assay,the activity of lactate dehydrogenase (LDH) was measured by a colorimetric method,and the levels of extracellular NO were measured by a nitrate reductive enzymatic method.The endothelial nitric oxide synthase (eNOS) mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results:Compared with the model group,liraglutide could significantly increase cell metabolic ability,reduce LDH release,increase NO release and eNOS mRNA expression (P<0.05 or P<0.01).The above effects of liraglutide were partly inhibited by glucagon like peptide-1 (GLP-1) receptor antagonist exendin (9-39)(P<0.05).Conclusion:Liraglutide can improve endothelial relaxation function in HUVECs in the state of hypoxia and high glucose in vitro.The effect might be related to up-regulating eNOS mRNA expression and promoting NO release.

Objective To compare the effect and safety of endoscopic duodenal papillal incision combined dilatation(ESBD) and endoscopic sphincterotomy(EST) in the treatment of common bile duct stone.Methods A retrospective analysis of 71 cases of choledocholithiasis,according to surgical methods,were divided into group EST (45 cases) and group ESBD (26 cases).The effect and safety of the above two kinds of treatment was evaluated through observing the rate of one-time stone clearance and postoperative complications.Results The success rate of removing calculous of the two groups was above 95%,and there was no significant difference between the two kinds of treatment(EST and ESBD)(P>0.05).Recent complications:the incidence rate of hemorrhage of the EST group was 0.0%,which of the ESBD group was 3.8%,there was no significant difference between the two groups(P>0.05).The incidence rate of postoperative pancreatitis of the EST group was 8.8%,which the ESBD group was 3.8%,there was no significant difference between the two groups(P>0.05).The diameter of the removed stone of the two groups:the EST group was (0.70±0.35)cm,and the ESBD group was (1.41±0.69)cm,there was significant difference between the two groups(P<0.05).Conclusion There are no significant differences in early complications and the success rate of removing stone between the EST group and the ESBD group.However,ESBD preserved teat sphincter's physiological barrier function to the greatest extent and the diameter of the removed stone was obviously larger than the EST group.Therefore,it is much safer for those who have quite large common bile duct stone or people who are with joint periampullary diverticulum to be treated by ESBD.

Objective To evaluate the clinical results of surgical treatment of isolated fractures of the sustentaculum tali of calcaneus via medial approach.Methods The data of 23 patients with isolated fractures of the sustentaculum tali of calcaneus was retrospectively analyzed who were treated with open reduction and internal fixation with cannulated screw or Kirschner wire via medial approach from September 2006 to March 2011.There were 19 males and 4 females,with an average age of 26.3 years (range,17-41 years).Associated injuries included 4 cases of talus fracture,4 of metatarsal fracture,and 3 of cuboid fracture.The functions of hiudfoot were assessed by American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores pre-operation and post-operation respectively.Results Fifteen patients got followed up with an average 20.5 months (10-56 months).Thirteen patients were rated as good,2 as excellent,and the excellent and good rate was 100％(15/15).All the fractures were stabilized reliably,and got clinical union with no obvious complications occurred.Time of fracture union was 8-10 weeks,with an average of 8.5 weeks.Three patients felt mild transient pain during the recovery of walking,but their pain disappeared quickly after physical therapy.No patients developed wound infection,nonunion and other complications.Conclusion For isolated fractures of the sustentaculum tali of caleaneus with articular surface displaced greater than 1 mm or involving the articular surface of middle subtalar joint,open reduction and internal fixation operation via medial approach under direct visualization is recommendable.

Objective To analyze the prevalence and genetic characteristics of influenza A(H3N2) viruses in the city of Xiangyang in 2022-2023, and to provide a scientific basis for predicting the epidemic and mutation of influenza virus. Methods Throat swab specimens of the influenza like cases were collected from national influenza monitoring sentinel hospitals in Xiangyang every week. RNA was extracted from the specimens for influenza diagnosing using real-time RT-PCR．Viruses were isolated from H3N2 positive specimens, and HA and NA genes were amplified and sequenced．3D modeling analyses were conducted. Results The gene phylogenetic tree showed that the H3N2 isolates in 2022-2023 belonged to 3C.2a1b.2a1 and 3C.2a1b.2a2 branches, respectively. The A(H3N2) influenza virus strains all had amino acid point mutation sites on important antigenic determinants of HA protein. The epitope mutations of the 2022 A(H3N2) strain mainly occurred in regions B, C, and D. The epitope mutations of the A(H3N2) strain in 2023 mainly occurred in regions C and D. Different glycosylation sites of HA gene were found in 2022-2023 strains. No variation was found in key amino acid sites associated with neuraminidase inhibitor resistance. The difference of overall structure was not obvious in the three-dimensional simulation structure diagram. Conclusion The A(H3N2) influenza strains isolated in this study have shown antigenic drift, especially the mutation of HA, which may affect the protective effect of the vaccine on the local population and lead to influenza epidemic. The variations of HA and NA suggest that close attention should be paid to the epidemic and genetic variation of H3N2 subtype influenza virus, to provide a scientific basis for the selection of influenza virus vaccine strains and the prevention and control of influenza.

Objective To study the serotype, drug resistance, and molecular typing characteristics of Salmonella enteritidis strains isolated from patients with diarrhea in Xiangyang City. Methods Serotyping, drug sensitivity test, pulsed field gel electrophoresis (PFGE) typing, and whole genome sequencing were carried out on Salmonella enteritidis strains isolated from diarrhea patients from 2019 to 2020. Results From the drug susceptibility test , Salmonella enteritidis in this region showed varying degrees of drug resistance, with 94.7% of Salmonella enteritidis strains being multidrug-resistant. The PFGE electrophoretogram of Salmonella enteritidis showed that 19 strains could be divided into 6 PFGE types by cluster analysis, among which 12 strains belonged to the epidemic dominant type. MLST typing showed that all 15 strains were ST11 type, carrying multiple drug resistance genes (mdsA, mdsB, mdsC, gols, and sdiA) and mdtK gene, and all carried various invasiveness-related virulence genes. However, there were slight differences in virulence genes related to the secretion system of the pathogenic island among the different strains. The phylogenetic tree analysis of the system evolution showed that 15 strains could be divided into 2 different evolutionary branches. Conclusion The situation of multidrug resistance in Salmonella enteritidis in Xiangyang is serious, with dominant PFGE types and ST types, and it carries multiple drug resistance genes and various invasiveness-related virulence genes.

Objective To understand the exposure level of disinfection by-products (DBPs) in drinking water in Xiangyang City and to evaluate the health risks of the population, so as to provide reference for the safety risk control of drinking water. Methods Sampling and laboratory testing were conducted according to national standard methods. The risk assessment model provided by USEPA was used to evaluate the health risks. Results Trichloromethane (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM) and tribromomethane (TBM) were detected in drinking water in Xiangyang City. The contents of halogenated hydrocarbon DBPs in water disinfected with liquid chlorine were higher than those with chlorine dioxide. The contents of three DBPs were higher in wet season than those in dry season except TBM. The content of TCM in pipe network terminal water and secondary water supply disinfected with chlorine dioxide was higher than that in factory water. The carcinogenic risk of DBPs in drinking water disinfected with liquid chlorine was 4.33×10^-5, and the non-carcinogenic risk was 0.114. The carcinogenic risk of DBPs in drinking water disinfected with chlorine dioxide was 1.24×10^-6, and the non-carcinogenic risk was 3.15×10^-3. Conclusion The health risk of DBPs in drinking water in Xiangyang City is acceptable, but TCM produced by liquid chlorine disinfection should be paid more attention. It is recommended that chlorine dioxide disinfection be used to reduce the health risks of halogenated hydrocarbon DBPs.

Objective To investigate vitamin K3 (VK3) effect on apoptosis of human liver cancer cells and its mechanism. Methods The SMMC-7721 cells were cultured in the experiment. The inhibitory effects of VK3 on SMMC-7721 cells were tested by CCK-8. Morphological evaluation of apoptosis was performed by Hcechst33342 staining. The distribution of cell cycle and apoptosis, and the expression of Fas were assayed by flow cytometry. Fas mRNA expression were detected by RT-PCR. And the concentration of soluble Fas(sFasL) in the culture supernatant were measured by EIJSA. Results The inhibitory rates ofVK3 (at concentrations of 2,5,10,20,25 umol/L for 48 h) on SMMC-7721 growth were 33.8% ,50.1%,63.9% ,78.5% and 84.7%, respectively. Compared with the control group, the number of cells in the G0/G1 phase increased, while that of S phase decreased. The apoptotic cell rates were 18.75%, 25.80%,38.80% ,29.92% and 26.18% ,respectively. The apoptosis cells were strongly stained by Hcechst33342.On exposure to VK3 at the concentration of (2,5,10 umol/L) after 48 h, the mean fluorescence intensity ofFas on cell surface and the expression of fas mRNA and the concentration of FasL in the culture supematantin SMMC-7721 were increased, but they all decreased at the high concentration of VK3. Conclusion VK3can inhibit the proliferation of SMMC-7721 cells and induce apoptosis via up-regulating expression of Fas and sFasL.

Objective: To replace esophageal defects with artificially composed biodegradable materials and non-biodegradable materials. Met hods: A two-layered tube consisting of a collagen-chitosan sponge and an inner polyurethane stent was used to replace 5 cm esophageal segmental defect s in 15 dogs. The inner polyurethane stent was removed endoscopically at weekly intervals from 2 or 4 weeks. Results: Partial regeneration of es ophageal epithelia was observed in 5 dogs at week 2, and progressing constricti on occurred and the dogs became unable to swallow within 1 month. In the 10 dog s that the polyurethane stent was removed at week 4, regenerated esophageal tiss ue successfully replaced the defects, and complete epithelization was observed 1 month after surgery. Complete regeneration of esophageal mucosa structures, inc luding mucosal smooth muscle and mucosal glands were observed 3 months after surgery, and partial regeneration of esophageal muscle tissue was also observed 6 months after surgery. Conclusion: Our artificial prosthesis i n reconstruction of the cervical esophagus segment in dogs is feasible. Through temporary polyurethane tube, collagen-chitosan sponge provides a three-dimensi onal structure suitable for the regeneration and sufficient degradation time for the complete regeneration of esophagus.

Objective To investigate the expression of interferon-induced protein 44 (IFI44) gene in the leukocytes of the peripheral blood samples from patients with systemic lupus erythematosus (SLE), and to evaluate the relationship between the expression level and disease activity. Methods Mononuclear cells in the peripheral blood samples from 100 SLE patients were compared with those of 40 disease controls and 40 healthy donors (HD) and the expression of the IFI44 was evaluated by quantitative real-time PCR.Comparisons between groups were performed with ANOVA, and the correlation analysis between the level of expression was higher in SLE patients than disease controls and healthy donors (26.8±5.3, 7.4±2.7, 5.2±2.0,respectively) (P=0.0012, P=0.005), but no difference was found between disease controls and healthy donors. Mild disease activity and the SLE patients with stable disease (63.1±22.4, 28.0±7.2, 9.2±1.8, respectively)and 24 hours urine protein level (r=0.42, P=0.000). Conclusion IFI44 is demonstrated to be highly expressed in SLE patients. The level of IFI44 may be a promising candidate biomarker for identifying SLE activity.