AIM: To determine whether chronic hypercholesterolemia affects ionic currents on cardiac ventricular myocytes of rats. METHODS: Whole - cell patch - clamp technique was used to record the ionic currents in single cardiac myocytes isolated from normal cholesterolemia and hypercholesterolemia rats. RESULTS: In the hypercholesterol group (group Ⅱ ), serum total - cholesterol level was significantly higher than that of normal group (group Ⅰ) [ (3. 10 ±tricular myocytes of rats, 50% repolarization of action potential duration (APD50) prolonged from (70. 86 ± 8.12) ms (group Ⅰ) to (116.16±6.90)ms (group Ⅱ) (n=10 in each group, P＜0.01); APD90 prolonged from (95.10±7. 27)ms (group Ⅰ ) to (144. 04 ± 7.39)ms (group Ⅱ ) (n = 10 in each group, P ＜ 0. 01 ); at the test potential of - 120 mV, Ik1 increased from ( - 16. 98 ±4. 54) pA/pF(group Ⅰ ) to ( - 19.92 ±4.08) pA/pF (group Ⅱ ) (n = 12 in each group, P ＜ 0. 05 ); at the test potential of 0 mV, ICa- L decreased from ( - 8.56 ± 1.29) pA/pF ( group Ⅰ ) to ( -5. 24 ± 0. 90) pA/pF ( group Ⅱ ) ( n = 10 in each group, P ＜ 0. 01 ); at the test potential of + 60 mV, Ito decreased from (13.20±1.97) pA/pF (group Ⅰ) to (10.30±1.97) pA/pF (group Ⅱ) (n=8 in each group, P＜0. 05). CON-CLUSION: Hypercholesterolemia affects the ionic currents on cardiomyocytes of rats greatly, which may be the ionic mechanism of cardiac toxicity induced by hypercholesterolemia.

Objective To explore comparison between methotrexate and uterine arterial embolization in β-HCG, bleeding volume and success rate of women with cesarean scar pregnancy after cesarean section.Methods 42 patients who were diagnosed with cesarean scar pregnancy after cesarean section were collected.All patients were randomly divided into uterine arterial embolization group and methotrexate group,21 cases in each group corresponding treatment were given respectively, after the treatment, the serum levels of β-HCG, bleeding volume and success rate were detected in all patients.Results After treatment, compared with methotrexate group, the serum level ofβ-HCG was lower in the uterine arterial embolization group,and the difference was statistically significant(P<0.05); the bleeding volume was lower in the uterine arterial embolization group(P<0.05); the success rate was higher in the uterine arterial embolization group(P <0.05).Conclusion Compared with methotrexate,the uterine arterial embolization can significantly reduce the serum level ofβ-HCG in patients with cesarean scar pregnancy after cesarean section,reduce the amount of bleeding, improve the success rate of treatment.

Objective:The abstraction of characteristics in CT perfusion image is crucial in the clinical diagnosis of brain infarction and cerebral tumor.Methods:After describing the relevant parameters of CT cerebral perfusion imaging,such as cerebral blood flow (CBF),cerebral blood volume (CBV),the mean transition time (MTT) and the permeability surface (PS).With using Visual C ++ programming and CT scan images sequence obtained,the time-density curves of the brain tumor region were visualized.The more accurate hemodynamic parameters were calculated through an improved algorithm basing on the deconvolution and measuring method of the length,angle,area and other parameters.Conclusion:The quantitative analysis of feature parameter is of great benefit to clinical doctor in making timely diagnosis and treatment of brain tumor.

Objective To clarify the role of FLT3 signaling-dependent pulmonary conventional dendritic cells (cDCs) in the pathogenesis of lipopolysaccharide (LPS)-induced acute lung injury (ALI),and as well as the modulation effects of cDCs in vivo on the inflammatory responses to acute lung injury.Methods Thirty C57BL/6 male mice were divided into normal control group,LPS group,FLT3L pretreatment group,lestaurtinib,(a high efficient and specific blocker in FLT3 signal pathway) pretreatment group and vehicle (DMSD) control group.FLT3L and lestaurtinib were administrated subcutaneously for 5 days.Murine model of ALI was subsequently established by intra-tracheal application of LPS and lung specimens were harvested 6 h or 24 h later.The accumulation and maturation of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate lung inflammation.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Lung myeloperoxidase (MPO) activity was measured to evaluate neutrophil infiltration.Transcription factors Tbet/GATA-3 mRNA ratio was determined to estimate balance of Th1/Th2 response.IFN-γ and IL-4 were quantified to evaluate Th1-specific and Th2-specific cytokine production respectively.Results The accumulation and maturation of pulmonary cDCs peaked at 6h after LPS challenge.FLT3L pretreatment significantly stimulated the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to markedly deterioration of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/GATA-3 mRNA ratio were elevated (P ＜ 0.05).Furthermore,the production of IL-6,TNF-α and IFN-γwas markedly increased by FLT3L pretreatment (P ＜ 0.05).In contrast,lestaurtinib pretreatment markedly inhibited the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to significant improvement of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/ GATA-3 mRNA ratio were decreased (P ＜ 0.05).Furthermore lestaurtinib efficiently suppressed the production of IL-6,TNF-α and IFN-γ (P ＜ 0.05).Conclusion This study thus demonstrated that FLT3 signaling-dependent pulmonary cDCs could control the initiation of acute lung inflammation response to LPS-induced ALI through the regulation of neutrophil infiltration and balance of Thl/Th2 response.

Objective To study the phenotype and function of CD4 +CD25 + Treg cells in the peripheral blood of patients with systemic sclerosis (SSc) and their relationship with fibrosis.Methods The proportion of Foxp3, CD127, CTLA-4 and CD69 on CD4+CD25+ Treg cells in peripheral blood were detect by flow cytometry;the levels of TGF-[1 and IL-10 in serum were detect by enzyme-linked immunosorbent assay (ELISA) in patients with SSc.The correlation between Treg cells and the score of chest HRCT, MRSS, and disease activity was analyzed.T test and Pearson correlation analysis were used for statistical analysis.Results ① Compare to the control group, the proportion of CD4+CD25+ Treg cells in peripheral blood of SSc patients was increased significantly(12.9±2.4 vs 14.9±2.2, t=2.63, P=-0.012), and the expression of CD69+, CTLA-4+ on CD4+CD25+ Treg cells was decreased significantly (P＜0.01).② Compare to the control group, the proportion of CD4+CD25+Foxp3 + cells and CD4+CD25+CDI27-cells in peripheral blood of SSc patients was increased significantly (respectively, 3.3±0.7 vs 5.0±0.7, 5.1±1.6 vs 7.6±2.0, t=7.03, 4.195;P＜0.01), but no correlation between them was detected.③ The level of TGF-β1 in the serum of the SSc patients was lower than that of the control group(86±29 vs 133±29 ng/ml, t=-5.026, P=0.000).However, IL-10 had no significant difference between the two groups.④ The proportion of CD4+CD25 +Foxp3 + cells and CD4~D25 +CD127-cells in peripheral blood of SSc patients was positively correlated with the scores of chest HRCT (respectively, r=-0.541, P=0.02;r=0.486, P=0.041), and no correlation was observed with ESR, CRP.In addition, CD4+CD25+Foxp3+ cells were associated with MRSS.Conclusion The proportion of CD4+CD25+ Treg cells in the peripheral blood of SSc patients is increased, but they alters the immune function.The different phenotypes of Treg cells of CD4+CD25+Foxp3+ cells and CD4+CD25+CD127-cells in peripheral blood of SSc patients are increased significantly, which changes along with skin and lung fibrosis.The associated cytokine TGF-β1 is reduced, and IL-10 is not significantly changed.

BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.

Objective:To explore the biological function of miR-132 in ovarian cancer and the target. Methods: 22 cases ovarian cancer tissue and non-tumor tissue adjacent were collected,the expression of miR-132 in tumor tissue and non-tumor tissue, normal ovarian epithelial cells and ovarian cancer cell were detected by RT-PCR. The normal ovarian epithelial cells which the expression of miR-132 maximum or minimum were chosen, and they were divided into two groups, respectively with transfection of negative control plasmid ( NC) and miR-132 mimic plasmid. The expression of miR-132 after transfection was detected by RT-PCR,the cell proliferation and cell apoptosis were detected by CCK-8 method and flow cytometry instrument respectively,the expression of Ezrin protein was detected by Western blot. Results:The expression of miR-132 in tumor tissue was significantly lower than the tumor tissue adjacent,the expression of miR-132 in ovarian cancer cell lines was significantly lower than normal ovarian epithelial cells, the differences were statistically significant (P<0. 05). The SKOV3 cell lines was chosed for gene transfection,compared with NC group, transfection with miR-132 mimic plasmid could significantly reduce cell proliferation, increase cell apoptosis, the difference had statistical significance ( P<0. 05 ) . Western blot results showed that up-regulation miR-132 significantly increased the Ezrin protein expression in ovarian cancer SKOV3 cells ( P<0. 05 ) . Conclusion: In ovarian cancer, miR-132;inhibits proliferation and induces apoptosis of ovarian cancer via Ezrin,it may be a tumor suppressor gene.

Objective To evaluate the effect of sucralfate and acid-suppressive drugs on preventing ventilator-associated pneumonia (VAP) in mechanically ventilated patients.Methods All randomized controlled trials (RCTs),which studied the effect of sucralfate and acid-suppressive drugs on the incidence of VAP in mechanically ventilated patients,were searched from PubMed,Embase and the Cochrane Library during January 1966 to March 2013 via manual and computer retrieval.All related data were extracted.Meta analysis was conducted using the statistical software RevMan 5.2 and the quality of the RCTs was strictly evaluated with the methods recommended by the Cochrane Collaboration.Results A total of 15 RCTs involving 1315 patients in the sucralfate group and 1568 patients in the acid-suppressive drug group were included in this study.The incidence of VAP was significantly reduced in the sucralfate group (RR =0.81,95％ CI 0.7-0.95,P =0.008),while no difference was found between the two groups in the incidence of stress-related gastrointestinal bleeding (RR =0.96,95％ CI 0.59-1.58,P =0.88).No statistical difference was found in the days on ventilator,duration of ICU stay and ICU mortality in the two groups (all P values ＞ 0.05).Conclusion In patients with mechanical ventilation,sucralfate could decrease the incidence of VAP,while has no such effect on the stress-related gastrointestinal bleeding,the days on ventilator,duration of ICU stay and ICU mortality.

Objective To investigate the relationship between serum inflammatory cytokines and atherosclerosis in type 2 diabetic patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods One hundred and three patients with type 2 diabetes were divided into OSAHS group (OSAHS group) and type 2 diabetes mellitus group (non-OSAHS group) according to the results of polysomnography(PSG),and 35 healthy subjects were selected as control group.Serum levels of total cholesterol,triglyceride,fasting plasma glucose(FPG),glycosylated hemoglobin(HbA1c),tumor necrosis factor-α(TNF-α),high-sensitivity C-reactive protein (hs-CRP) and carotid intima-media thickness (IMT) were measured,and their relationship with OSAHS were analyzed.Results The waist circumference,BMI,FPG,HbA1c,IMT,TNF-α and CRP in OSAHS group were significantly higher than those in non-OSAHS group (P＜0.05).Logistic regression analysis showed that,IMT was independently associated with TNF-α and hs-CRP.Conclusion Patients of T2DM with OSAHS have poor blood glucose control and higher incidence of atherosclerosis.High levels of TNF-α and hs-CRP may be involved in the formation of atherosclerosis and plaque occurrence and development.

The effect of troglitazone on glucose-stimulated insulin secretion (GSIS) in pancreatic β-cells and its mechanism were investigated.10 μmol/L troglitazone had no effect on basal insulin secretion,but significantly decreased GSIS and stimulated AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) phosphorylations (all P<0.01).These reactions were completely reversed by AMPK inhibitor compound C,suggesting that the troglitazone acutely inhibits insulin secretion via stimulating AMPK activity in beta cells.