To facilitate the expression of GL 7 ACA acylase gene in a recombinant E coli , a fragment of the gene, in which the signal peptide was deleted by PCR method, was inserted into a prokaryotic expression vector, pET 28a By colony PCR method screening, a recombinant plasmid pET ACY was obtained and then transformed into the expression host BL21 (DE3) The influences of induction conditions such as IPTG concentration, the time of induction and the induction temperature on the expression of the recombinant protein were investigated Under optimal condition, the enzyme activity could reach 266 U/L Finally, the recombinant GL 7 ACA acylase can be easily isolated to a purity of about 80% by a simple anion ion exchange chromatography with enzyme activity recovery of 50%

Objective: To examine and analyze the nm23, p16 gene expression in gastric cancer tissure,and follow up patients 5 years to discuss the relationship between nm23 and p16 gene synergy expression and gastric cancer biological characteristic and prognosis.Methods: nm23 and p16 protein in gastric cancer tissue and control were detected by immunohistochemistry,and the patients had been followed up for 5 years. Results: In 84 samples of gastric cancer, nm23 positive rate was 46.43%, p16 was 44.05%, the positive rate of gastric cancer tissue and metastasitic lymph node was lower than that of normal control, normal tissue near cancer or benign polyp,and these two genes were related to the depth of tumor invasion and clinical stage.The mortality and recurrence-metastasis rate was higher in these low expression group, and had a shorter median survive period. Conclusion: Abnormal expression of nm23 and p16 gene plays a important role in gastric cancer recurrence and devolepment and may be one of markers for evaluating tumor biological behavior and prognosis.

Adipose tissue is not only involved in energy metabolism, but also recognized as an important endocrine organ.With declining ovarian function, endogenous estrogen levels decrease, leading to body fat mass accumulation and centripetal redistribution in postmenopausal women.Furthermore, energy metabolism, adipokine levels, local estrogen synthesis and the expression of estrogen receptors in adipose tissue also undergo changes, which may induce adipose tissue dysfunction.Changes in body fat mass and fat distribution and adipose tissue dysfunction are associated with increased risk of metabolic diseases and cardiovascular diseases in postmenopausal women, which have a negative impact on their health and quality of life.In this paper, the impact of menopause on body fat mass, regional distribution and adipose tissue functions and the underlying mechanisms are reviewed.

Purpose: This study aimed to identify the risk factors and sonographic variables that could be integrated into a predictive model for endometrial cancer (EC) and atypical endometrial hyperplasia (AEH) in women with abnormal uterine bleeding (AUB). Materials and Methods: This retrospective study included 1837 patients who presented with AUB and underwent endometrial sampling. Multivariable logistic regression was developed based on clinical and sonographic covariates [endometrial thickness (ET), resistance index (RI) of the endometrial vasculature] assessed for their association with EC/AEH in the development group (n=1369), and a predictive nomogram was proposed. The model was validated in 468 patients. Results: Histological examination revealed 167 patients (12.2%) with EC or AEH in the development group. Using multivariable logistic regression, the following variables were incorporated in the prediction of endometrial malignancy: metabolic diseases [odds ratio (OR)=7.764, 95% confidence intervals (CI) 5.042–11.955], family history (OR=3.555, 95% CI 1.055–11.971), age ≥40 years (OR=3.195, 95% CI 1.878–5.435), RI ≤0.5 (OR=8.733, 95% CI 4.311–17.692), and ET ≥10 mm (OR=8.479, 95% CI 5.440–13.216). :A nomogram was created using these five variables with an area under the curve of 0.837 (95% CI 0.800–0.874). The calibration curve showed good agreement between the observed and predicted occurrences. For the validation group, the model provided acceptable discrimination and calibration. Conclusion The proposed nomogram model showed moderate prediction accuracy in the differentiation between benign and malignant endometrial lesions among women with AUB.

Crohn's disease(CD)is a chronic non-specific intestinal inflammatory disease,and the incidence of perianal fistulizing CD(PFCD)is 17%-43%. Non-cutting setons is the first choice for surgical treatment of PFCD. Some new surgical methods are effective for specific types of PFCD,however,the efficacy of most new methods remains to be confirmed by further studies. The multidisciplinary team(MDT)mode has become a new direction of PFCD surgery. This article reviewed the advances in surgical treatment of PFCD.

Objective To compare the short-term and long-term outcomes between laparoscopic and open total proctocolectomy with ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC).Methods The retrospective cohort study was conducted.The clinical data of 150 UC patients who underwent total proctocolectomy with IPAA in the Renji Hospital of Shanghai Jiaotong University between January 2003 and December 2016 were collected.Among 150 patients,87 undergoing laparoscopic total proctocolectomy with IPAA and 63 undergoing open total proctocolectomy with IPAA were respectively allocated into the laparoscopy group and open group.Observation indicators:(1) comparisons of intra-and post-operative situations;(2) postoperative complications;(3) follow-up situation.Follow-up using outpatient examination and telephone interview was performed to detect postoperative recovery and complications up to December 2017.Measurement data with normal distribution were represented as x-±s and comparison between groups was analyzed using the t test.Comparison between groups of count data was analyzed using the chi-square test.Results (1) Comparisons of intra-and post-operative situations:operation time,volume of intraoperative blood loss,time of initial intestinal stoma exsufflation and duration of hospital stay were respectively (306±3) minutes,(197± 12) mL,(62.1±1.8) hours,(8.2±0.4) days in the laparoscopy group and (224±4) minutes,(308±24) mL,(75.6±2.0) hours,(10.1±0.6) days in the open group,with statistically significant difference between groups (t =16.23,4.33,5.03,2.61,P＜ 0.05).All patients discharged successfully from hospital.All the 150 patients underwent stoma reversion of ileum at 3-12 months postoperatively,and the average time in the laparoscopy group and open group was respectively (6.0±5.6) months and (6.0±4.6)months,with no statistically significant difference between groups (t =0.01,P＞ 0.05).(2) Postoperative complications:cases with postoperative wound infection,retention of urine and frequency of defecation ＞ 4 times / day were respectively 2,8,21 in the laparoscopy group and 8,15,29 in the open group,with statistically significant differences between groups (x2 =5.25,4.37,0.96,P＜0.05).The cases with postoperative intestinal obstruction,anastomotic leakage,pelvic infection,pouch infection,pouch-related Crohn's disease and hyperplasia of ileal pouch were respectively 3,10,5,23,2,1 in the laparoscopy group and 8,7,4,24,1,0 in the open group,with no statistically significant differences between groups (x2=3.65,0.11,0.01,0.96,0.17,0.82,P＞0.05).Patients with postoperative complications were improved by acid suppression,fasting,anti-infection and fluid infusion.(3) Follow-up situation:150 patients were followed up for 12-60 months,with a median time of 48 months.There was no abnormality of postoperative anastomotic stoma and intestinal mucosa through comparison of colonoscopy results between pre-operation and 5 year postoperatively.During the follow-up,50 patients had shapeless stool and irregular defecation (times ＞ 4 times / day) at 3 years after stoma reversion of small intestine bypass,including 21 in the laparoscopy group and 29 in the open group,with a statistically significant differences between groups (x2 =4.72,P＜0.05).Eleven and 10 patients in the laparoscopy group and open group had shapeless stool and irregular defecation at 5 years postoperatively,but status were improved compared with the preoperative status,with no statistically significant difference between groups (x2=0.32,P ＞ 0.05).Conclusion The security of laparoscopic total proctocolectomy with IPAA for UC is equivalent to that of open total proctocolectomy,with the better short-term and long-term outcomes.

Objective To investigate the extraction and purification methods of serum specific endothelial cell antibody of renal transplant recipients with rejection after renal transplantation.Methods Human umbilical vein endothelial cell (HUVEC)was isolated and cultured.The serum samples of the renal transplant recipients with poor renal function after renal transplantation were collected.Specific endothelial cell antibody was screened out with flow cytometry;antibodies against human leukocyte antigen (HLA)and major histocompatibility complex class Ⅰ-related chain A (MICA)were detected by Luminex platform.After the existence of specific endothelial cell antibody in the serum sample was confirmed,specific endothelial cell antibody was absorbed with HUVEC.The cell was washed and then the absorbed antibody was eluted from the cell membrane.Antibody IgG in the eluent was purified and concentrated again with Protein-A /G magnetic beads.Antibody activity in the eluent was detected by flow cytometry and the purified specific endothelial cell antibody (IgG)was identified by SDS-polyacrylamide gel (SDS-PAGE)and Western blot.Results In the serum of 386 renal transplant recipients,the serum samples of 5 renal transplant recipients with serum creatinine (Scr) >400 μmoI /L,negative anti-HLA antibody,negative anti-MICA antibody and median fluorescence intensity (MFI) >16 were selected.Purified specific endothelial cell antibody IgG showed immunoglobulin heavy chain (purity > 95%)by SDS-PAGE gel.Flow cytometry showed that the purified antibody had the feature of rebinding with the surface antigen of vascular endothelial cell.Conclusions The purification method of using human umbilical vein endothelial cell to absorb specific endothelial cell antibody in the serum of renal transplant recipients may obtain good effect.

Objective:To investigate the effect of actin related protein 2/3 complex subunit 2 (ARPC2) gene silencing on the biological characteristics of papillary thyroid carcinoma (PTC) TPC-1 cells through lentivirus-mediated RNA interference.Methods:TPC-1 cells infected with nonsense short hairpin RNA (shRNA) sequence lentivirus (shCtrl) was used as the control group. TPC-1 cells infected with ARPC2 shRNA interference sequence lentivirus (shARPC2) was used as the experimental group, in which the expression of ARPC2 gene was specifically interfered. The effects of silencing the expression of ARPC2 gene on the proliferation of TPC-1 cells were detected by using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot and colony formation test. Flow cytometry and Western blot were conducted to detect the effect of silencing ARPC2 gene on TPC-1 cells apoptosis and related proteins.Results:shARPC2 could efficiently infect TPC-1 cells, and the expression efficiency of green fluorescent protein was over 85%. Compared with the control group, TPC-1 proliferation was inhibited in the experimental group. The ratio of S-phase cells in the experimental group was reduced compared with that in the control group [(14.79±0.21)% vs. (21.13±0.33)%, t = 27.77, P < 0.05]. The ratio of G 1 and G 2/M-phase cells in the experimental group was increased compared with that in the control group [G 1 phase: (67.57±0.08)% vs. (62.06±0.36)%, t=25.56, P < 0.05; G 2/M phase: (17.64±0.12)% vs. (16.91±0.17)%, t=6.154, P < 0.05]. Meanwhile, the expressions of cell cycle-related proteins CDK2, CyclinE and CyclinD were reduced in the experimental group. The number of clone formation in the experimental group was less than that in the control group, the difference was statistically significant [(10±2) vs. (161±6), t=9.011, P < 0.05]. In addition, the apoptotic ratio of cells in the experimental group was higher than that in the control group [(8.60±0.77)% vs. (4.08±0.40)%, t=9.011, P < 0.05]. Western blot showed that the expressions of anti-apoptotic factors p21 and bcl-2 were reduced in the experimental group, while the expression of pro-apoptotic factor bax was increased. Conclusion:The interference with the expression of ARPC2 regulated by shRNA can inhibit the proliferation, and promote the apoptosis of PTC TPC-1 cells, indicating that ARPC2 may be a possible biological new target for the treatment of PTC.

Objective:To investigate the application value of barbed suture in reinforcing anastomosis during laparoscopic radical resection of rectal cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 215 patients with rectal cancer who underwent laparoscopic radical resection in the Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine from March 2016 to December 2018 were collected. There were 122 males and 93 females, aged (62.3±0.7)years, with a range from 20 to 75 years. Of 215 patients, 86 patients undergoing laparoscopic radical resection of rectal cancer with barbed suture for anastomosis were allocated into barbed suture group, and 129 patients undergoing laparoscopic radical resection of rectal cancer without reinforced anastomosis were allocated into traditional group, respectively. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) follow-up. Follow-up using outpatient examination and telephone interview was performed to detect postoperative complications, anastomotic healing and patency of patients up to May 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were described as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Ordinal data were analyzed using the Friedman non-parametric test. Results:(1) Intraoperative situations: patients in the two groups underwent laparoscopic radical resection of rectal cancer successfully, without conversion to open laparotomy. The volume of intraoperative blood loss, operation time, distance between the anastomosis and dentate line, length of auxiliary incision of the barbed suture group were (127±9)mL, (160.2±3.8)minutes, (3.56±0.15)cm, (4.12±0.11)cm, respectively, versus (114±6)mL, (128.9±2.4)minutes, (3.67±0.12)cm, (4.25±0.09)cm of the traditional group. There was a significant difference in the operation time between the two groups ( t=7.33, P<0.05), but no significant difference in the other indicators between the two groups ( t=1.32, 0.61, 0.94, P>0.05). (2) Postoperative situations: the time to first flatus after surgery and duration of postoperative hospital stay of the barbed suture group were (72.5±2.2)hours and (8.1±0.5)days, respectively, versus (76.2±1.7)hours and (8.0±0.5)days of the traditional group, showing no significant difference between the two groups ( t=1.33, 0.87, P>0.05). (3) Follow-up: 215 patients were followed up for 6-12 months, with a median follow-up time of 8 months. In the barbed suture group, 2 patients had postoperative anastomotic leakage, 5 had urinary retention, 6 had incision infection, 4 had intestinal obstruction, 3 had bowel dysfunction in 6 months after operation. In the traditional group, 13 patients had postoperative anastomotic leakage, 15 had urinary retention, 11 had incision infection, 8 had intestinal obstruction, 9 had bowel dysfunction in 6 months after operation. There was a significant difference in the anastomotic leakage between the two groups ( t=4.77, P<0.05), but no significant difference in the urinary retention, incision infection, intestinal obstruction, bowel dysfunction in 6 months after operation between the two groups ( t=2.07, 0.17, 0.22, 1.26, P>0.05). Patients in the two groups showed unobstructed intestinal tract on enteroscopy. Conclusion:The barbed suture for reinforcing anastomosis in radical resection of rectal cancer is safe and feasible, which can reduce the incidence of anastomotic leakage.