Objective Bone morphogenetic protein 4 ( BMP4) induces rat myocardial hypertrophy in H9c2 cells, but the spe-cific mechanism remains unclear .The study aimed to elucidate the role of autophagy in myocardial hypertrophy and its relationship with extracellular signal regulating kinase ( ERK1/2) signal transduction pathway . Methods H9c2 cardiomyocytes were randomly divided into 4 groups:control group, BMP4 group, BMP4+PD98059 (ERK1/2 signal pathway inhibitor) group and BMP4+3MA (autophagy inhibitor) group.With PD98059(50μmol/L) and 3MA(5mmol/L) blocking for 30min, BMP4 (50μg/L) were added.Expressions of tiny tube related proteins 1 light chain 3 (LC3) and p-ERK1/2 were detec-ted after culturing for 30min.48h later, measurements were made on cell surface area , average protein content and α-smooth muscle actin (α-SMA ) protein expression level .Cell morphology and size were measured respectively by inverted microscope and Image J software .Total protein content was detected by BCA , and western blot was used to measure the expressions of LC3, ERK1/2, p-ERK1/2 and α-SMA. Resul ts 30min later, the expressions of LC3 and p-ERK1/2 were significantly higher in BMP4 group than in control group [(1.54 ±0.05) vs (1.95 ±0.11),(0.94 ±0.04) vs (1.33 ± 0.06),P<0.01] and no significant difference was found in other two groups .Compared with BMP4 group, significant decrease was found in BMP4+PD98059 and BMP4+3MA [(1.95 ±0.11) vs (1.59 ±0.08), (1.36 ±0.02);(1.33 ±0.06) vs (0.87 ±0.05), (0.95 ±0.15), P<0.01], while no significant difference was found between BMP 4+PD98059 group and BMP4+3MA group.48h lat-er, the cell area, protein content and α-SMA protein expression level are significantly lower in control group than in BMP 4 group [(644.1 ±15.01)μm2 vs (745.6 ±14.43)μm2,(240.0 ±7.26)pg/cell vs (347.1 ±5.4)pg/cell,(1.22 ±0.06 vs 1.99 ±0.03),P<0.01], while the corresponding indexes were significantly higher in BMP 4+PD98059 and BMP4+3MA compared with BMP4 group [(745.6 ±14.43)μm2 vs (645.0 ±12.63)μm2, (647.7 ±13.89)μm2;(347.1 ±5.4)pg/cell vs (239.7 ±3.39)pg/cell , (241.1 ± 8.56)pg/cell;(1.99 ±0.03) vs (1.13 ±0.05);(1.12 ±0.02), P<0.01].No significant difference was found as to the indexes be-tween BMP4+PD98059 group and BMP4+3MA group(P>0.05). Conclusion Autophagy may participate in BMP4-induced rat myo-cardial hypertrophy in H9c2 cells through ERK1/2 signal transduction pathway .The clinical treatment of myocardial hypertrophy may benefit from the blocking of autophagy or ERK 1/2 signal transduction pathway .

Objective To establish a quantitative SYBR GreenⅠreal-time PCR method for detection of Metadherin (MTDH) gene expression level in breast cancer, and to investigate the relationship between MTDH mRNA expression level and the clinicopathological characters. Methods Real-time PCR was employed to determine the expression level of MTDH mRNA in peripheral blood of 80 specimens of breast cancer patients and normal females. Results In the 80 specimens, MTDH mRNA was positively expressed in the peripheral blood of 61 cases of breast cancer patients while negatively expressed in the 19 cases of normal females. Among the 61 breast cancer patients, MTDH mRNA showed high expression in 34 cases, accounting for 55.7%, and showed low expression in 27 cases, accounting for 44.3%. Both of the differences in expression rate has statistic significance (Ratio = 2.02 ± 0.81, P<0.05). MTDH expression relative to GAPDH expression in the peripheral blood of breast cancer patients was 1.15 ± 0.36. MTDH mRNA expression has no connection with age, estrogen and progesterone receptors , as well as HER-2.(P＞0.05). There is statistical difference for MTDH mRNA expression level between the group with lymph node metastasis and the group without lymph node metastasis (Ratio=2.02 ± 0.81,P<0.05). MTDH mRNA expression level changed between clinicopathological stage I, II and III, IV. Conclusion The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of MTDH mRNA, which may be closely related to the occurrence and development of breast cancer.

Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.

Objective To analyze the current situation and influence factors of ordinary medical college undergraduates' contact with scientific research at early stage in order to provide references for scientific research.Methods Totally 1940 students majoring in clinical medicine,imaging,traditional Chinese medicine and nursing (2008 -2010 grade) in China Three Gorges University were enrolled to do questionnaine and SPSS 17.0 was used to do statistical analysis.Results Totally 1653copies of questionnaires were collected from 1940 students,the recovery rate was 85.21％.Two hundred and nineteen students ( 13.25％ ) participated in scientific research,65.28％ students thought college propaganda to be ordinary,95.43％ students got benefits from scientific research.The main influence factors of scientific research were lack of time (23.73％),insufficient knowledge reserves (22.03％) and researchers' own problems (39.73％).Conclusions Medical school should expand the range of scientific research and strengthen propaganda.Medical students should arrange research time and constantly improve their comprehensive ability so as to achieve good results.

Objective To investigate the expression of survivin and bcl-2 in human squamous cell carcinoma (SCC) lesions and cell line SCL-1. Methods Tissue samples from 60 patients with SCC and 10 normal human controls were immunohistochemically stained to detect the expressions of survivin and bcl-2.Western blot was used to measure the expressions of bcl-2 and survivin proteins in HaCaT human keratinocytes and SCL-1 human squamous cell carcinoma cells. Results In normal control tissues, there was no expressions of survivin or bcl-2, while in SCC, the expression rates of bcl-2 and survivin were 70% and 60%, respectively,and there was no statistical correlation between the expressions of bcl-2 and survivin (P >0.05). Neither the expression of survivin nor that of bcl-2 was correlated to patients' age, gender or lesional site (all P >0.05). A statistical correlation was observed between the pathological stage in patients and expression of bcl-2 as well as between lymph node metastasis and expression of survivin (both P < 0.05). Western blot analysis revealed a significant increase in the expression of survivin and bcl-2 in SCL-1 cells compared with HaCaT cells. Con-clusion In SCC, survivin and bcl-2 seem to play their roles via different anti-apoptotic pathways.

Objective To investigate the in vitro effect of interferon-γ (IFN-γ) and ATRA on the morphological transition, proliferation of and apoptosis in a human cutaneous squamous cell carcinoma cell line SCL-1. Methods Cultured SCL-1 cells were divided into 6 groups to be treated with ATRA of 1 μmol/L, various concentrations ( 100, 500, 1000 U/ml) of IFN-γ, the combination of ATRA of 1 μmol/L and IFN-y of 1000 U/ml,respectively, or to remain untreated. MTT assay and flow cytometry were performed to evaluate the cell proliferation and apoptosis. The morphological features of apoptotic cells were observed by a transmission electron microscope (TEM) and inverted phase contrast microscope after 1% propidium iodide staining. Results IFN-γ could inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and the most pronounced inhibitory effect was observed at a dose of 1000 U/ml . ATRA and IFN-γ induced an apoptosis in SCL-1 cells, and the early apoptosis rate was 4.84%, 11.96% and 18.71% in SCL-1 cells after treated with ATRA of 1 μmol/L, IFN-γ of 1000 U/ml and their combination, respectively. A series of morphological changes characteristic of apoptosis,such as bipolar changes, were observed in SCL-1 cells treated with ATRA and IFN-γ, with the presence of many early apoptotic cells, which showed a trend towards benign differentiation. Conclusions Within a certain concentration range, IFN-γcan promote the differentiation, but inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and ATRA could enhance the effects of IFN-γ.

Objective To analyze the influence of hepatosteatosis on pancreatic β-cell function in type 2 diabetes mellitus (T2DM).Methods A total of 213 subjects with T2DM from Metabolic Disease Hospital,Tianjin Medical University from January 2013 to December 2013 were included in the study.Non-alcoholic fatty liver disease (NAFLD) was diagnosed with abdominal ultrasonography.Patients were divided into two groups:T2DM with NAFLD and T2DM without NAFLD.ALT,AST,γ-glutamyltransferase,serum lipid,glycosylated hemoglobin A1 c (HbA1c),fructosamine,fasting glucose,insulin and 2 hours plasma glucose,insulin after 75g glucoseload were detected.The insulin resistance and β-cell function were assessed by HOMA-IR and HOMA-β.Results Among the 213 T2DM subjects,51％ (108 cases) were with NAFLD.The HOMA-IR [4.76 (2.83,7.21) vs 2.79 (1.76,4.37),P ＜ 0.05] and HOMA-β [49.18 (37.78,85.09) vs 29.50 (18.09,45.54),P ＜ 0.05] were significantly higher in T2DM with NAFLD than those in T2DM alone.Within subjects with T2DM and NAFLD,the HOMA-IR [6.28 (2.87,8.17) vs 2.95 (2.07,3.66) P ＜ 0.05] and HOMA-β [59.18 (37.78,85.09) vs 30.59 (28.56,34.49),P ＜ 0.05] levels were higher in subjects with normal liver function than those with abnormal liver function.Conclusions T2DM patients with NAFLD have severer insulin resistance than those without NAFLD.The β-cell function of those patients was compensatory increased,which was decreased in subjects with abnormal liver function.

Objective To evaluate the safety and efficacy of colour doppler in guiding the percutaneous nephrolithotomy(PCNL) for Staghom calculi.Methods The clinical records of 46 patients with renal calculi who underwent PCNL were retrospectively analyzed.Patients' mean age was 43 years old,and the range of diameter of stone was 3.0 -7.5 cm.Among these cases,29 cases had complete staghorn calculus.One case had isolated kidney stone.And the other 6 patients had open surgery history.Using Colour Doppler guidance,the percutaneous nephrolithotomy for renal calculi was conducted.Results F22 percutaneous channel was successfully established in 55 sides of 46 patients,with the first and the second phase surgeries of 41 and 14 sides respectively.Single-,double- and three-channel PCNL were performed in 38,16,and 1 sides respectively.Abdominal x-ray conducted at 3 -4 days post operation revealed residual stones on 18 sides,with the range of 0.4 -2.0 cm in diameter.Second-phase lithotripsy was conducted on 14 sides of patients.After the first and the second phases of surgery,4 sides having residual stones ranging 0.6 - 1.0 cm,underwent extracorporeal shock wave lithotripsy.Ater the third-phase surgery,the diameters of residual stones were much less than 0.5 cm and patients were treated with medication.The total rate of clearance was 87.0％ (48/55).The duration of surgery was 65 - 160 minutes and 95 minutes on average.One patient having delayed bleeding was cured with selective renal artery embolization.There were no complications such as nephrectomy,deaths,pleural or intestinal damage during the period of study.Patients were followed up for 3 to 18 months.Six of 9 patients with renal insufficiency recovered after surgery.The serum creatinine in the remaining 3 patients ranged 185 -220 μmol/L Conclusion Colour Doppler-guided percutaneous nephrolithotomy for renal calculi is safe and effective.

The related substances in docetaxel injection were identified by LC-MS/MS. Ethyl acetate was used to extract the injection to remove the pharmaceutical excipients. HPLC separation was carried out on a Hedera ODS-2 column (150 mm x 2.1 mm, 5 microm) with a mobile phase consisting of acetonitrile - 0.1% acetate acid aqueous solution (40: 60). Electrospray ionization source was set in the positive mode for the LC-ESI-MS/MS, and the ion monitoring modes were full scan and product ion scan. According to the mass spectra of the related substances, the fragment profiles were explained, and the chemical structures were elucidated. Docetaxel and its main related substances were well separated. Nine related substances in docetaxel injection were detected by LC-MS/MS. Their chemical structures were proposed, and four of them were identified in the docetaxel injection for the first time. The established LC-MS/MS method is effective in the separation and identification of the related substances in docetaxel injection. The test results are useful for its quality control.

Objective To investigate the effects of silencing TSG-6 gene modified bone marrow mesenchymal stem cells (BMSC) transplantation on liver allotransplantation rejection in rats.Methods BMSC and KCs were isolated from rats and cultured in complete medium.We down-regulated TSG-6 expression of BMSC with lentiviral vectors carrying short hairpin RNA (LV3-shTSG-6).TSG-6mRNA and protein level of BMSC were tested respectively by quantitative real-time PCR and western blot analysis.After co-cuhured between BMSC and KCs,the expression of TNF-α and TSG-6 in cell supernatants were tested.Then,we established the orthotopic liver transplantation models in rats,the rats of each group were killed at 1 days,3 days and 7 days after operation.The serum levels of AST,ALT,TBIL,γ-GGT,TNF-α,IL-6 and IL-4 were tested with ELISA in each group,TSG-6mRNA and protein level of liver tissues obtained from each group were tested by quantitative real-time PCR and western blot analysis.The 1iver tissues of each group were stained with HE,then microstructure of liver tissues were observed under light microscope.The postoperative survival rates in other rats of each group were observed.Results The lentivirus transfection efficiency of mesenchymal stem cells was beyond 70 percent;After co-cultured between BMSC and KCs,the expression of TSG-6 in TSG-6-shRNA-BMSC + KCs group supernatant was significantly lower in different time point than that in BMSC + KCs group and TSG-6-NC-BMSC + KCs group (P ＜0.05),and the expression of TNF-α peak in BMSC + KCs group and TSG-6-NC-BMSC + KCs group supernatants were at 6 hours,which were significandy lower than those at 12 hours (P ＜0.05),but the expression of TNF-α in TSG-6-shRNABMSC + KCs group at 12 hours was significantly higher than those in BMSC + KCs group and TSG-6-NC-BMSC +KCs group (P ＜0.05);After transplantation,the serum levels of AST,ALT,TBIL,γ-GGT,TNF-α and IL-6 in TSG-6-shRNA-BMSC group was significantly higher than those in TSG-6-NC-BMSC group and BMSC group in different time point (P ＜ 0.05),but had no significant difference compared with PBS group;The serum levels of IL-4 in TSG-6-shRNA-BMSC group,TSG-6-NC-BMSC group and BMSC group was significantly higher than that in PBS group(P ＜ 0.05),but TSG-6-shRNA-BMSC group compared with TSG-6-NC-BMSC group and BMSC group,the serum levels of IL-4 was significantly lower (P ＜ 0.05);In pathological changes,we found that the degree of liver rejection in TSG-6-shRNA-BMSC group and PBS group were seriously obvious,and were graded Ⅱ-Ⅲ with Banff schedule;Comparation of postoperative survival time in each group,TSG-6-shRNA-BMSC group and PBS group were (16.6 ±4.6) d and (15.4 ± 6.7) d respectively,which were signifcantly lower than those in TSG-6-NCBMSC group (69.6 ± 28.1) d and BMSC group (69.2 ± 28.2) d (P ＜ 0.05).Conclusion Transplantation of BMSC secreted TSG-6 could,to some extent,mitigate acute liver transplantation rejection.