Objective To investigate the pathological and electro-physiological process of terminal sprouting after muscle injection of Botulinum toxin type A(BTXA) in rats. Methods Sprague-Dawley rats were randomly divided into a BTXA group and a control group. Five unit of BTXA was injected into the right gastrocnemius of the rats in BTXA group, instead of 0.9% saline in the control group. The morphologic axon analysis and SFEMG (mean consecutive difference and fiber density) were measured at 1,4,8,12 week after injection. Results The mean consecutive difference was prolonged in 1-4 week, and got better at 8 week, recovered at 12 week. It was revealed of the recovery of neuromuscular junction at 12 week after BTXA injection. The results of fiber density and morphologic axon analysis demonstrated terminal sprouting after injection of BTXA. Conclusion Motor nerve terminal sprouting could appear after BTXA injection and the function of neuromuscular junction could become recovered at 12 week.

Objective To investigate the radiobiological effects of VPA-BSANPs on C6 and U87 glioma cells in vitro.Methods C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h and 24 h,and MTT assay was used to determine cell viability.C6 and U87 cells were treated with different concentrations of VPA and VPA-BSANPs conbined with X-ray irradiation (0,2,4,6,and 8 Gy),and colony formation assay was used to determine plating efficiency (PE).C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h,followed by X-ray irradiation (0,4,and 8 Gy),and flow cytometry using Annexin V-FITC/PI staining was used to examine cell apoptosis.Western blot was used to evaluate the effects of VPA and VPA-BSANPs on radiation-induced apoptosis protein expression.One-way ANOVA was used for comparison of means with homogeneity of variance between multiple groups,and the t-test was used for comparison of means between two groups.Results Without irradiation,VPA and VPA-BSANPs had no significant inhibitory effects on the proliferation of C6 and U87 cells (P=0.328,0.920).The PE of cells treated with VPA-BSANPs combined with irradiation was significantly lower than that of cells treated with VPA combined with irradiation (P=0.000).In C6 and U87 cells,VPA-BSANPs combined with irradiation increased the expression of p53 and Bax (P =0.000,0.000 and P =0.010,0.002),but reduced the expression of Bcl-2 (P =0.008,0.000).Active caspase-3 fragments were only found in the cells treated with VPA-BSANPs combined with irradiation and VPA combined with irradiation,but were less in the former cells than in the latter cells (P=0.004).The active fragments of peroxisome proliferator-activated receptor were only found in the cells treated with VPA-BSANPs combined with irradiation.Conclusions VPA-BSANPs can increase the radiosensitivity of C6 and U87 glioma cells in vitro,possibly by promoting the apoptosis of tumor cells induced by radiation.

Objective To assess the efficacy of fish oil in the treatment of IgA nephropathy using the method of Meta-analysis. Methods Randomized controlled trials of fish oil in the treatment of IgA nephropathy were searched in the database of Cochrane library,PubMed, EMBASE and CNKI. Data extracted from the literatures were analyzed with Revman software (version 5.0). Results In comparison with the controlled group, proteinuria in the fish oil group was significantly decreased [SMD=-0.27, 95%CI (-0.52 to -0.03), P=0.03], while the renal function deteriorated [SMD=0.30,95%CI(0.05 to 0.55), P=0.02]. Conclusion Fish oil can decrease the proteinuria of IgA nephropathy, but can not prevent renal function from deterioration.

Objective To analyze the therapeutic effect and prognosis of peritoneal dialysis in patients with end-stage polycystic kidney disease.Methods A retrospective analysis was performed on patients with polycystic kidney disease who were treated with peritoneal dialysis for more than 3 months between July 2007 and September 2016 in the Second Hospital Affiliated to Soochow University.A total of 45 patients were enrolled in this study.Another 45 patients of non-diabetic nephropathy were selected as the control group matched by gender,age,and time of PD initiation.The information of the two groups such as general data,dialysis related complications,incidence of peritonitis,prognosis was recorded.Survival analysis was performed using the Kaplan-Meier method and Log-rank test.The risk factors affecting patients' survival were analyzed with Cox regression model.Results There were no significant difference in pre-dialysis age,sex ratio,blood pressure,urine volume,body weight,eGFR,biochemical data,and the proportion of hypertension and diabetes mellitus in the polycystic kidney group and control group.24 h ultra-filtration volume,4 h D/Pcr,Kt/V and Ccr between the two groups showed no significant difference (all P ＞ 0.05).The incidence of peritonitis and the time of the first peritonitis in the two groups respectively as one episode per 82.4 months vs one episode per 81.5 months,(35.8±22.8) months vs (34.5±20.9) months had no statistical difference.The ratio of hernia (6.6％ vs 2.2％),thoracic and abdominal leakage (4.4％ vs 2.2％),dialysate leakage (0 vs 0),catheter dysfunction (4.4％ vs 6.6％),exit-site infections (11.1％ vs 6.6％),tunnel infections (4.4％ vs 2.2％) and non PD related infections (11.1％ vs 13.3％) had no significant difference.The 1-year,3-year,5-year patient survival of two groups respectively were 95.2％ vs 93.3％,78.9％ vs 75.0％,67.6％ vs 64.9％ (P=0.475),and 5-year technique survival was 78.7％ vs 76.7％ (P=0.623),demonstrating no obvious difference.Cox regression analysis showed that age and serum albumin were risk factors for the survival of patients.Conclusions The effect and prognosis of peritoneal dialysis in patients with polyeystic kidney and non polyeystic kidney were similar.Peritoneal dialysis is not the contraindication of polycystic kidney.Peritoneal dialysis can be used as a routine renal replacement therapy in patients with polycystic kidney disease.

Objective To explore ability of the vpr gene of human immunodeficiency virus type 1 ( HIV-1 vpr) to induce cell G_2 arrest and apoptosis, and the influence when it mutated, the relationship between Vpr-induced G_2 arrest and apoptosis inductions. Methods Fourteen mutant vpr fragments selected from Chinese patients with HIV. Both eukaryotic expression vector pcDNA3.1( + ) and PCR products purified, double-cut by Hind Ⅲ and BamH Ⅰ and the cut products legated and transformed into competent cells JM109. The 14 reconstructed plasmids electronically transfected into Jurkat-cells, and established cells with pcDNA3. 1-vpr , pcDNA3. 1-vpr-Fs and pcDNA3. 1 blank cells, and without pcDNA3. 1 cell. Cells were harvested after 24 h. mRNA expression was detected by RT-PCR, the DNA content and percentage of apoptosis were monitored by flow cytometry. Results Transfected with 14 mutant HIV-1 Vpr protein, cells display different G_2 percentage and apoptosis ratio. HIV-1 vpr induce cell cycle G_2 arrest and apoptosis, wherase Vpr Fs with a C-terminal end truncation, vector pcDNA3.1( + ) and the blank cells can not. The G_2 percentage and apoptosis ratio reduced when transfected with vpr expressing mutating of 70V, 85P, 86G, 94G compared to the wild type. Subtype AE has a weaker potential to induce cell cycle G_2 arrest and apoptosis. Preliminary, we find that the higher G_2 percentage followed the higher ratio of apoptosis. Conclusion HIV-1 vpr can induce cell cycle G_2 arrest and apoptosis, wherase Vpr Fs with a C-terminal end truncation can not. We firstly found that mutated sites of 70V, 85P, 86G, 94G may reduce the ability of Vpr to induce cell cycle G_2 arrest and apoptosis, subtype AE of vpr in Chinese HIV-1 patients has a weaker potential to induce cell cycle G_2 arrest and apoptosis. Analysis of various mutations in the vpr gene revealed that the extent of Vpr-induced G_2 arrest correlated with the levels of apoptosis. And investigate the pathegenesis of HIV vpr. This can also make a good foundation for further study on gene therapy.

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.

Objective To investigate the relationship of red cell distribution width (RDW) with all-cause mortality and cardiovascular disease (CVD) mortality in patients undergoing maintenance hemodialysis (MHD).Methods A retrospective analysis was performed in patients who initiated MHD from January 2008 to September 2017 in the hemodialysis center of the Second Affiliated Hospital of Soochow University.Basic data on demographic,dialysis and laboratory were collected,and echocardiography indicators and clinical outcomes were recorded.Patients were divided into four groups according to the quartile of RDW level.Kaplan-Meier survival analysis was used to compare the difference of survival rate among the groups.Cox regression analysis was used to analyze the risk factors of all-cause and CVD-related mortality,and predictive value of RDW for all-cause and CVD-related death in hemodialysis patients.Results A total of 268 MHD patients were enrolled in this study with age of (60.9± 15.8) years and dialysis duration of (58.1±9.1) months,including 159 males (59.3％).Kaplan-Meier survival analysis showed that the 1-year overall survival rates of Q1 group (RDW≤ 13.8％,n=61),Q2 group (RDW 13.9％-14.6％,n=66),Q3 group (RDW 14.7％-15.6％,n=73)and Q4 group (RDW≥15.7％,n=68) were 96.8％,95.1％,93.1％ and 85.7％ respectively;3-year overall survival rates were 88.5％,87.5％,59.2％ and 51.8％ respectively;5-year overall survival rates were 71.5％,65.4％,33.6％ and 17.7％ respectively;The difference between the groups was statistically significant (all P ＜ 0.01).The 1-year CVD survival rates were 98.4％,96.6％,95.8％ and 92.4％ respectively;3-year CVD survival rates were 94.8％,92.5％,84.4％ and 70.4％ respectively;5-year CVD survival rates were 86.9％,81.3％,65.6％ and 51.3％ respectively;The difference between the groups was statistically significant (all P ＜ 0.01).Multivariate Cox regression analysis showed that RDW≥15.7％ was an independent risk factor for all-cause and CVD-related mortality in MHD patients.The risk of all-cause mortality in Q4 group was 3.098 times higher than that in Q 1 group (95％ CI 1.072-8.950,P=0.037) and the risk of CVD-related mortality was 2.661 times (95％ CI 1.111-8.342,P=0.048).Receiver operating characteristic curve (ROC) showed that RDW=14.85％ was the best cut-off point for predicting the all-cause mortality in HD patients (P ＜ 0.01),RDW=15.45％was the best cut-off point for predicting the cardiovascular disease mortality (P ＜ 0.01),and RDW=14.45％ had a higher 5-year survival rate (P ＜ 0.01).Conclusion RDW can independently predict all-cause and CVD-related mortality risk in hemodialysis patients,and it has important value for prognosis.

Objective: To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability. Methods: A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels. Results: Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment. Conclusion TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Objective:To explore the relationship between the hepatic caudate lobe boundary and the ductal system so as to guide the identification of the anatomical relationship during liver surgery.Methods:The specific parts were observed and the liver parenchyma was removed according to 41 cadaveric liver autopsy specimens. The critical relationship between the hepatic caudate lobe and other ducts was observed to explore the reticular duct structure.Results:The plane formed by the hepatic hilar plate and Arantius ligament served as the boundary between the caudate lobe and other hepatic lobes. The caudate lobe hepatic portal vein was composed of numerous small branches from its left and right branches. The portal vein adjacent to the vena cava was mainly derived from the left branch, and to a lesser extent from the right branch. Blood was drained straight from the caudate lobe vein into the inferior vena cava via the short hepatic vein. There were three or four bile duct branches in the caudate lobe. The main source of arterial blood flow were the left and right branches of the hepatic artery. An avascular zone of loose connective tissue was found between the caudate lobe and the retrohepatic inferior vena cava.Conclusion:The hepatic caudate lobe is an independent lobe. During hepatic caudate lobe surgery, the plane formed by the hepatic hilar plate and Arantius ligament can serve as the boundary between the caudate lobe and other hepatic lobes and be used for anatomical site identification. The duct system of the caudate lobe's is complicated, but it also has its own distinct regularity.

Objective To investigate the correlation between oxidative stress and pain after laparoscopie myomectomy.Methods 84 patients with uterine myoma who were treated in Changsha Hospital of traditional Chinese Medicine from March 2014 to March 2017 were enrolled in the study.The patients were divided into the control group (n =42) and the observation group (n =42) according to the random number table method.The patients in the control group were treated with conventional laparotomy hysteromyoma expetion,and the patients in the observation group were treated by laparoscopic myomectomy.The levels of oxidative stress indexes [reactive oxygen species (ROS),malondialdehyde (MDA),advanced oxidation protein products (AOPP)] and pain indexes [interleukin-6 (IL-6),substance P (SP),prostaglandin E2 (PGE2) and visual analogue scale (VAS) score] were compared between two groups before operation,1 d after operation and 3 d after operation.Pearson correlation analysis was used to analyze the correlation between oxidative stress and pain.Results The levels of plasma ROS,MDA and AOPP in the two groups 1 d after operation and 3 d after operation were higher than those before operation (all P ＜ 0.05),the levels of plasma ROS,MDA and AOPP in observation group 1d after operation and 3 d after operation were significantly lower than in the control group (all P ＜ 0.05).The levels of serum IL-6,SP,PGE2 and VAS score in the two groups 1 d after operation and 3 d after operation were all higher than those before operation (all P ＜ 0.05),the levels of serum IL-6,SP,PGE2 and VAS score in observation group after 1 d and 3 d after operation were all lower than those in control group (all P ＜ 0.05).The results of Pearson correlation analysis showed that ROS,MDA and AOPP was positively correlated with PGE2 (P ＜ 0.05).Conclusions Laparoscopic myomectomy can effectively reduce the oxidative stress response to postoperative hysteromyoma and relieve postoperative pain,and oxidative stress is positively correlated with PGE2 after operation.