Objective To explore the role and mechanism of tumor necrosis factor-α (TNF-α) in the formation of chronic transplant renal interstitial fibrosis and to study Smurf2 expression change and role played in this process.Methods We collected 26 cases of normal renal tissues and 26 cases of renal allograft specimens from chronic allograft dysfunction patients to observe the degree of renal interstitial fibrosis by Hematoxylin and eosin (HE) staining and Masson staining.Immunohistochemical staining was applied to detect the expression and distribution of E-cadherin,α-smooth muscle actin (α-SMA) and TNF-α,Smad ubiquitination regulatory factor 2 (Smurf2).Furthermore,The HK2 cells were divided into five groups depending on different concentrations of TNF-α (0,10,20,50,and 100 ng/mL).After 48 h,we collected the cells to analyze the expression changes of Smurf2,E-cad,α-SMA by Western blotting.Results As compared with normal group,the degree of renal interstitial fibrosis in CAD group was aggravated according to the results of HE and Masson staining (P＜0.01).Immunohistochemical staining results showed that the positive expression of E-cadherin was reduced,and that of α-SMA,TNF-α and Smurf2 increased greatly in CAD group as compared with normal group (P＜0.01).The results of Western blotting also revealed that after treatment with different concentrations of TNF-α for 48 h,the expression of E-cadherin was downregulated,and that of α-SMA and smurf2 was up-regulated in HK2 cells (P＜0.05).Conclusion TNF-α may promote the formation of allograft renal interstitial fibrosis via transdifferentiation of human tubular epithelial cells to mesenchymal cells by up-regulating the expression of Smurf2.

Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10%FBS, during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation, including three-drug combination group and four-drug combination group. The protocol of mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination induced differentiation, four-drug combination was successfully achieved in over 90% of the cell inducing, which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.

Objective To evaluate endoscopic pancreaticobiliary separation (EPBS) in patients with gallstone and occult pancreaticobiliary reflux (OPBR).Methods The clinical data of 47 cases with gallstone and OPBR from Oct 2013 to Oct 2016 was analyzed retrospectively.Results The mean gallbladder bile amylase (GBA) was (864 ± 575) U/L.40 cases have undergone the treatment of endoscopic retrograde cholangiopancreatography.26 cases were diagnosed as duodenal papillitis,16 cases as periampullary diverticula,14 cases as long nipple,5 cases as atrophic papilla,3 cases as ampulla stone and 2 cases as papillary tumor.9 were diagnosed as pancreaticobiliary maljunction in the 14 patients with long nipple.33 cases were performed with EPBS.GBA was tested in 16 cases,and the GBA of 15 cases returned to normal level.The difference of GBA was statistically significant [(1 161 ±764) U/L vs.(47 ± 17) U/L,(t =5.641,P ＜ 0.05)].Patients were followed up for 1 to 4 years,among 36 cases without cholecystectomy,there was no recurrence of cholecystolithiasis after EPBS in 27 cases,but 2 cases had recurrent gallstones in 9 cases without EPBS,and the difference of gallstone recurrence rate was statistically significant (x2 =21.340,P ＜ 0.05).Conclusions Pancreaticobiliary junction diseases is an important cause for gallstone formation and OPBR.EPBS can avoid pancreaticobiliary reflux and reduce the recurrence rate of gallstone after choledochoscopic lithotomy.

Objective: To study the coping styles and its relationship with quality of life among part of the female breast cancer patients in Shanghai in 2014. Methods: In August of 2014, 1-3 block groups affiliated to Shanghai Cancer Rehabilitation Club were selected from each of the 17 districts of Shanghai by convenient sampling method. Respondents were recruited from these block groups via putting up posters in the community or top three hospitals nearby. The inclusion criteria were as follows: female, aged from 18 to 80 years old; the initial diagnosis or primary tumor was breast cancer, and active treatments including surgery, radiotherapy and chemotherapy were finished; capable of basic reading and comprehension, and there was no communication disorder; daily activities were not limited, and with no mental disorder or dysgnosia. Totally 2 205 respondents were included. Questionnaire survey was conducted to collect the information of demographic characteristics, disease characteristics, result of European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30), and result of the Ways of Coping Inventory-Cancer Version (WOC-CA). 1 968 valid questionnaires were withdrawn. The results of respondent characteristics and WOC-CA were under descriptive analysis, and the differences of coping styles among groups of different demographic characteristics were compared using t test. In addition, we analyzed the correlation between respondents' coping styles and quality of life using partial correlation analysis. Results: The average age of the 1 968 respondents was (58.7±7.4) years old, and BMI was (24.4±5.4) kg/m². The score of positive coping styles was 2.04±0.41, and the score of negative coping styles was 1.79±0.33. The scores of Physical Functioning (PF), Role Functioning (RF), Cognitive Functioning (CF), Emotional Functioning (EF), Social Functioning (SF), and Global Health (GH) were 83.40±12.18, 90.80±15.92, 80.72±16.29, 84.67±15.19, 80.99±20.91, 65.27±21.21, respectively. Positive coping styles correlated with PF, EF, and GH positively, and the correlation coefficients were separately 0.581 (P=0.046), 0.593 (P=0.045), 0.770 (P<0.001). Negative coping styles correlated with RF, CF, EF, and SF negatively, and the correlation coefficients were separately-0.672 (P=0.021),-0.815 (P=0.005),-0.121 (P<0.001),-0.123 (P<0.001). Conclusion Part of the female breast cancer patients in Shanghai in 2014 mainly adopted positive coping styles, and in general the positive coping styles correlated with quality of life positively and negative coping styles correlated with quality of life negatively.