Objective To explore the risk factors and prevention measures for anastomotic leakage after rectal radical resection.Methods The clinical data of 404 patients with rectal radical resection were analyzed retrospectively and the risk factors for anastomotic leakage were analyzed.Results Thirty-one patients (7.67％,31/404) were subjected to anastomotic leakage.The mean leakage time was 6.5 (3-14) d postoperatively.The muhivariate Logistic regression analysis showed that preoperative hemoglobin (OR =3.023,95％ CI:1.101-8.303,P=0.031 8),tumor size (OR =2.543,95％ CI:1.075-6.018,P=0.033 7) and tumor distance from anal verge (OR =3.160,95％ CI:1.387-7.199,P=0.006 2) were the risk factors for anastomotic leakage.Conclusions Preoperative hemoglobin,tumor size and tumor distance from anal verge are significant factors for anastomotic leakage.Therefore correction of anemia,improvement of surgical technique and suitable use of preventive diversion stoma ane all benefit for prevention of anastomotic leakage after rectal radical resection.

The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.
Animals ; Bone Morphogenetic Proteins ; genetics ; Chromosomes, Artificial, Bacterial ; DNA Transposable Elements ; Fibroblasts ; Goats ; genetics ; Keratins ; genetics ; Transfection

Objective To investigate the in vitro effects of bone marrow mesenchymal stem cells (BM MSCs) on the replication of HBV with the participation of lymphocytes and to analyze the possible mechanism.Methods The HBV genomic DNA transfected HepG2.2.15 cell line was used to evaluate the HBV replication.Bone marrow and spleen samples were collected from BN rats for the isolation of BM MSCs and T lymphocyte cells, respectively.Five groups of co-culturing with different cells were designed in this study.The cellular activities of lymphocytes and HepG2.2.15 cells were detected at the time of 24 h, 48 h and 72 h after co-culturing by using MTT method.The levels of HBV DNA and HBV cccDNA were detected by real-time polymerase chain reaction ( PCR) .T cell subsets in co-culture were measured by using fluores-cence labeled antibodies and flow cytometry analysis.ELISA was used to detect the levels of cytokines in the supernatant of cultured cells.Results Compared with HepG2.2.15 cells group, BM MSCs+HepG2.2.15 cells and splenic lymphocytes+HepG2.2.15 cells co-culture groups, the levels of HBV DNA and HBV cccDNA were significantly decreased in splenic lymphocytes+BM MSCs+HepG2.2.15 cells co-culture group after 48 h of culture [ HBV DNA: ( 181.000 ±14.731 ) IU/ml vs ( 6270.000 ±300.450 ) IU/ml, (2564.000±231.058) IU/ml, (2433.300±302.379) IU/ml;HBV cccDNA: (4.330×105 ±0.464×105 ) IU/ml vs (11.100×105±0.375×105) IU/ml, (8.930×105±0.778×105) IU/ml, (9.850×105±0.810× 105) IU/ml;P<0.01].The secretion of IFN-γin the supernatant of co-cultured cells was negatively corre-lated with HBV DNA level, but the levels of IL-10 and IL-22 were positively correlated with HBV DNA.The ratio of CD4+/CD8+cells was increased in splenic lymphocytes+BM MSCs+HepG2.2.15 cells co-culture group.The percentage of CD8+cells showed a positive correlation with HBV DNA.Conclusion BM MSCs could inhibit the expression of HBV DNA to enhance the clearance of HBV strains.It might be possibly due to rebalancing of Tc1/Tc2 cells and regulating the expression of autocrine agents and cytokines.

Objective: To analyze the epidemiological characteristics of latent syphilis in Yancheng City from 2016 to 2020, so as to provide insights into syphilis control. Methods: All reported cases with latent syphilis in Yancheng City from 2016 to 2020 was collected from the Communicable Disease Report System of China Disease Prevention and Control Information System, and the prevalence of latent syphilis was estimated and standardized by the seventh population census data in Yancheng City. The trends in the incidence of latent syphilis were evaluated using annual percent change (APC), and the temporal, regional and human distributions of latent syphilis patients were descriptively analyzed. In addition, the spatial clusters of latent syphilis incidence were identified using spatial autocorrelation analysis. Results: A total of 7 790 cases with latent syphilis were reported in Yancheng City from 2016 to 2020, and the standardized incidence of latent syphilis increased from 15.35/105 in 2016 to 28.70/105 in 2020 (APC=17.54%, t=5.357, P=0.013). Latent syphilis cases were reported in each month, and no obvious seasonable characteristics were seen. During the period from 2017 to 2020, the highest incidence of latent syphilis was seen in residents at ages of 70 to 79 years, with incidence rates of 41.71/105, 43.04/105, 75.79/105 and 72.94/105, respectively, and most cases were farmers (4 711 cases, 60.47%). The three highest incidence of latent syphilis was reported in Funing County (191.40/105), Tinghu District (137.13/105) and Yandu District (126.23/105). There was a positive spatial correlation of latent syphilis incidence in Yancheng City from 2016 to 2020 (Moran's I=0.23, Z=4.457, P=0.001), and two high-high clusters were identified in 14 townships (streets) of Funing County, Binhai County, Tinghu District, Sheyang County and Yandu District and 3 low-low clusters in 7 townships (streets) in Jianhu County, Tinghu District, Dongtai City and Sheyang County. Conclusions The incidence of latent syphilis appeared a tendency towards a rise, and there were remarkable spatial clusters identified in latent syphilis incidence in Yancheng City from 2016 to 2020. The elderly people and farmers are at high risk of latent syphilis.

Objective To explore the effect and mechanism of heme oxygenase-1 (HO-1) gene modified bone marrow mesenchymal stem cells (BMMSCs) on rat reduced-size liver transplantation.Methods 50 male Brown Norway (BN) rats were used to prepare BMMSCs.Male Lewis and BN rats were 75,respectively.BN rats were randomly divided into model group (n =25),stem cell group (n =25) and combined group (n =25).Acute rejection models following 50％ reduced-size transplantaton were established in rats using two-cuff technique,1 ml of normal saline,BMMSCs suspension,or HO-1/BMMSCs suspension were injected immediately after surgery.Rats were executed at an instant,3rd,7th and 14th day after surgery to identify BMMSCs and HO-1/adenovirus infection efficiency.Evaluated hepatic pathology by HE staining.Liver function indexes were detected.Portal vein pressure on 7th day after surgery was detected.The levels of endothelin-1 (ET-1) and nitric oxide (NO) in serum were detected using ELISA.The expressions of ET-1 in liver were detected by immunohistochemistry staining and Western blotting.Results High purity BMMSCs were obtained and HO-1/BMMSCs were successfully infected.Compared with model group,liver tissue injury and rejection were alleviated in stem cell group and combined group,liver function was improved,and the combined group was superior to stem cell group.The portal vein pressure in model group,stem cell group,and combined group were 21.3±0.2 mmHg,11.2±0.2 mmHg,and 10.1±0.1 mmHg,respectively.The portal vein pressure in three groups showed a decreasing trend,difference was statistically significant (P＜0.05).On the 3rd,7th and 14th day after surgery,compared with model group,the expression levels of ET-1 and NO in the stem cell group and the combined group were decreased,and the combined group was significantly lower than stem cell group (P＜ 0.05).Conclusion HO-1/BMMSCs improved liver function and portal vein pressure after reduced-size liver transplantation in rats,and may play a protective role by regulating ET-1/NO expression.

Objective To study the effect of bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic mechanical perfusion (NMP) on biliary epithelial cells (BEC) after DCD donor liver transplantation in rats.Methods The third generation of BMMSCs and the BMMSCs modified by Ad/HO-1 (Ad/HO-1/BMMSCs) were cultured,identified and expanded in vitro.To establish a stable NMP system device in vitro.The DCD liver transplantation models were constructed in rats after cardiac ischemia for 30 minutes,220 SD recipient rats were randomly divided into sham operation group (S group,n=44) static cold storage (SCS group,n =44) group,and simple NMP group (P group,n =44),BMMSCs combined with NMP group (BP group,n =44) and BMMSCs modified by Ad/HO-1 combine with NMP group (HBP group,n =44),NMP group,BP group and HBP group were subjected to vitro perfusion for 4h.The group were taken at 0,1,7 and 14 days after transplantation and the relevant indicators were detected,n =6 in each group.The survival rate of the recipient rats,liver function and pathological changes of the bile duct were observed.The expression of cytokeratin 19 (CK19) protein in BEC was detected by immunohistochemistry and Western blot.Apoptotic biliary epithelial cells were detected by TUNEL staining and the expression of apoptosis-related protein caspase-3 was detected by immunohistochemistry.Results The survival time of HBP group was significantly prolonged for (5.6 ±0.8) d in SCS group vs.(18.4 ±2.0) d in NMP group,(20.5 ± 1.5) d in BP group,(82.5 ±3.2) d in HBP group,the differences were statistically significant (all P ＜ O.05).Compared with other groups,the HBP group and the BP group were significantly improved in liver function and biliary pathology,and the expression of CK19 protein in BEC was significantly increased [(0.81 ±0.02) in S group vs.(0.35 ±0.03) in SCS group,(0.47 ±0.02) in NMP group,(0.63 ± 0.02) in BP group,(0.77 ± 0.01) in HBP group on postoperative day (POD) 14],the differences were statistically significant (all P ＜ 0.05).The number of apoptosis and the expression of apoptosis-related protein caspase-3 in HBP group were significantly decreased [(10.0 ± 1.2) in S group vs.(57.3 ±5.5) in SCS group,(40.1 ±4.6) in NMP group,(32.0 ± 2.2) in BP group,(13.7 ± 3.1) in HBP group on POD 14],the difference was statistically significant (all P ＜ 0.05).Compared with the BP group,the protective effect of the HBP group was more obvious,and the difference was statistically significant (P ＜ 0.05).Conclusion By the method of the BMMSCs modified by Ad/HO-1 combined with NMP in vitro preservation of rat,DCD donor liver can significantly improve the effect of BEC on rats and the survival rate after liver transplantation.

Liver transplantation is the most effective therapeutic options for the patients at the advanced stage, but with the amount of transplant surgery increasing, margin donors are used for transplantation in the case of severe donor organ deficiency. However, the commonly used cold storage technique has poor preservation effect on margin donors, resulting in an increase in the incidence of complications after transplantation. The donated liver quality is one the most important factors for the patients long term survival, so there is an urgent need for a new type of organ preservation technology to preserve the margin donors in vitro. This paper summarized the current research on the ex-vivo preservation methods of liver and the new mechanical perfusion preservation methods.

Objective:To explore the role of heme oxygenase-1 (HO-1) modified bone marrow mesenchymal stem cells (BMMSCs) in improving hepatic microcirculation after reduced-size liver transplantation (RLT) in rats.Methods:BMMSCs were isolated and cultured in vitro by adherence method. Then HO-1/adenovirus (Adv) was transfected for constructing HO-1/BMMSCs. The "dual-sleeve" method was employed for establishing an acute rejection model after 50% RLT. Immediately post-operation, 1 ml normal saline (NS) and BMMSCs or HO-1/BMMSCs single cell suspension were injected. The changes of surviving rats were observed by parameters at Day 3/7/14 post-operation. Five rats were observed at each timepoint. The serum level of mitochondrial aspartate aminotransferase (mAST) was detected; Na+ -K+ -ATPase in transplanted liver was measured by chemical colorimetry; mitochondrial ultrastructural changes were observed under a transmission electron microscope. Portal vein pressure was detected by Power Lab at Day 7 post-operation; the expressions of endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS) and von Willebrand factor (vWF) in liver tissues were detected by Western blot. Liver histopathological changes were observed by hematoxylin & eosin stain. The expression of vWF was detected by immunohistochemistry and serum level of hyaluronic acid (HA) detected by enzyme-linked immunosorbent assay (ELISA).Results:HO-1/BMMSCs could significantly lessen the pathological injury and rejection of 50% reduced-size transplanted liver, improve mitochondrial damage and energy metabolism, promote the expression of eNOS, suppress the expression of iNOS, reduce portal pressure, up-regulate the expression of hepatic sinus vWF and HA degradation, protect hepatic sinusoidal endothelial cells (SECs) and ultimately improve hepatic microcirculation. And the differences were statistically significant as compared with NS/BMMSCs group ( P<0.05). Conclusions:HO-1/BMMSCs may play an important role in protecting rat liver by improving hepatic microcirculation during RLT.

Objective:To explore the correlation between cerebrovascular hemodynamic index (CVHI) accumulative score and subclinical arteriosclerosis indicators.Methods:A total of 27 184 cases were collected from the Health Management Center,the Third Xiangya Hospital,Central South University.Linear regression analysis was carried out to confirm the correlations between CVHI accumulative score and the modified Framingham stroke profile (FSP),as well as between CVHI accumulative score and cerebrovascular diseases (ICVD) scale.The correlation between CVHI accumulative score and brachial-ankle pulse wave velocity (baPWV),carotid plaque orcarotid intima-media thickness (CIMT) was analyzed by multifactor logistic regression model in 11 580 cases.Moreover,the correlation between CVHI accumulative score and microalbuminuria or serum cystatin C was performed by multifactor logistic regression model in 9 860 cases.Results:In this study,the people whose CVHI accumulative score was less than 75 accounted for 12.98％.The CVHI accumulative score was negatively related with the modified FSP score (r=-0.484,P＜0.01) or ICVD score (r=-0.455,P＜0.01).The multifactor logistic regression model found that the baPWV,carotid plaque,microalbuminuria and serum cystatin C were independent predictors for CVHI accumulative score.Conclusion:The CVHI accumulative score is correlated with the modified FSP score,ICVD score and indexes of subclinical arteriosclerosis (baPWV,carotid plaque,microalbuminuria and serum cystatin C).The CVHI accumulative score could be used as a tool for zero-level and primary prevention of cerebral stroke.

Objective To summarize the clinical characteristics of recipients of renal transplantation who used tacrolimus extended-release capsules,to optimize the postoperative immunosuppressive regimen,and provide reference for the therapeutic administration of imnmunosuppressive agents after renal transplantation.Methods 156 patients who had renal transplant in our center were divided into three groups according to the time of the change of the extended-release tacrolimus capsules,and the blood glucose and blood lipids of each group were analyzed.Results The longer the postoperative duration was,the higher proportion of new-onset diabetes cases (P =0.025).There was no significant difference among the three groups of immune induction regimens.The immnunosuppressive regimen was changed from MMF (68.8％ in G3 group) to MPA (72％ in G1 group).With the prolongation of postoperative time,the dosage of tacrolimus decreased gradually.The mean tacrolimus concentration in the 3 groups was significantly different (P＜0.001) as time went by.There was no significant change in the average daily dosage before and after the change.The trough value before and after the change in the first two groups was significantly different (P＜0.001).Conclusion The extended-release tacrolimus capsules could be used in different stages after renal transplantation.After the conversion of the extended-release tacrolimus capsules,the dosage of adjuvant is reduced,and blood concentration and creatinine level are more stablem which is a more optimized immunosuppressive regimen.