Objective To estimate the biological effects of leptin on human HaCaT keratinocytes and explore their molecular mechanisms.Methods Cell counting kit-8 (CCK-8) was used to evaluate the proliferation of cultured HaCaT cells treated with different concentrations of leptin for 24 and 48 hours.Some HaCaT cells were classified into four groups to remain untreated,be treated with leptin (100 μg/L) and piceatannol (a specific inhibitor of STAT3 phosphorylation) alone or in combination for 24 hours,respectively,followed by the evaluation of cell proliferation using CCK-8 kit.Flow cytometry was performed to assess cell cycle of HaCaT cells treated with leptin of 100 μg/L,Western blot to determine the phosphorylation level of Erk1/2 and STAT3 in HaCaT cells treated with leptin of 100 μg/L for different durations.Statistical analysis was done by Student's t-test for unpaired data using GraphPad Prism 5 software.Results The proliferation of HaCaT cells was accelerated to different degrees after treatment with leptin of 50 and 100 μg/L for 24 and 48 hours,and the accelerating effect was in a dose-dependent manner within 24 hours (r =0.9989,P ＜ 0.05).Piceatannol apparently inhibited the promotive effect of leptin on the proliferation of HaCaT cells.There was an obvious elevation in the percentage of cells at S phase ((57.70 ± 5.88)％ vs.(42.50 ± 7.55)％,P ＞ 0.05),but a significant decrease in that at G0/G1 phase ((39.70 ± 1.57)％ vs.(45.20 ± 1.44)％,P ＜ 0.05),with a significant increase in proliferation index (0.603 ±0.0157 vs.0.564 ± 0.0144,P ＜ 0.05) in HaCaT cells treated with leptin of 100 μg/L for 24 hours compared with the untreated controls.Western blot showed that leptin of 100 μg/L markedly enhanced the phosphorylation level of STAT3 in HaCaT cells.Conclusion Leptin may upregulate the proliferation of HaCaT cells through activation of STAT3 pathway.

Objective To analyze the changes in the expression of Toll-like receptors 4 (TLR4) and 7 on the surface of murine bone marrow-derived dendritic cells following dengue virus type 2 (DEN2) infection.Methods DEN2 NGC strain was infected into BALB/c suckling mice through intracranial injection and injected into C6/36 cells to induce the in vivo and in vitro proliferation of DEN2, respectively.RT-PCR was performed to identify DEN2 RNA.Reed-Muench method was used to determine the 50% tissue culture infective dose (TCID50) of DEN2.Dendritic cells (DCs) were prepared by stimulating bone marrow cells isolated form C57BL/6 mice with IL-4 and GM-CSF and then identified by flow cytometry.The prepared murine bone marrow-derived DCs were infected with DEN2 and observed with direct immunofluorescence assay.Dynamic changes in the expression of CD11c, CD86 and I-A/I-E molecules on DCs after DEN2 infection were detected by flow cytometry.Levels of DEN2 RNA and the expression of TLR4 and TLR7 at mRNA level were dynamically detected by real-time quantitative PCR.Results The TCID50 of DEN2 to C6/36 cells was 10-5.8.Murine bone marrow-derived DCs were acquired with a purity of 70% and could be infected with DEN2 in vitro.The percentages of CD86 and I-A/I-E molecules on the surface of DCs infected with 1×105 TCID50 of DEN2 were statistically different from those of the negative control group.Neither of the two groups showed a significant difference in the percentages of membrane molecules over time.However, the percentages of membrane molecules on DCs increased with increasing viral load.Compared with the negative control group, the levels of DEN2 RNA in infection groups were increased with increasing virus load, while the expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level was decreased with increasing viral load.Conclusion DEN2 infection promotes the maturation of DCs.Expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level decreases with increasing viral load, which suggests that TLR4 and TLR7 are closely related to viral infection and play a certain role in the pathogenesis of DEN infection.

Objective To explore the complication and its cause and treatment of pace markers implantation in older patient.Methods Among 205 older patients aged 70 to 90 years,103 were male,the others were female.Of these patients,162 were implanted with single chamber pace marker,43 with dual chamber pace markers.Results 21 patients happened complication(10.2%),the common complications related to operation were blood effusion and hematoma formation in pocket 12 cases,lead dislodgement 4 cases and infection with or without pocket rapture 2 cases.Occurrence of blood effusion was related to aspirin administration,lead dislodgement and infection with or without pocket rapture were related to operation.Conclusion To pay attention to ample preparation,close operation and strict observation can decrease the complication rate when pace marker is implanted.The serious result will be avoided by timely and effective treatment.The older patient with pace markers implantation is safe.

One hundred and thirty seven pafients diagnosed by coronary angiography were recruited in the study and divided into coronary heart disease group(CAD)and control group.Relationship between serum apolipoprotein M levels and the severity of stenosis of coronary arteries was analyzed.In covariate analysis,serum apolipoprotein M levels were significantly lower in CAD group than those in control group adjusted for sex,age,body mass index(BMI)(P＜0.05).By stepwise muhiple regression,serum levels of apolipoprotein M were correlated positively with BMI(r=0.65,P＜0.01),and correlated negatively with total cholesterol(TC)(r=-0.53,P＜0.01)and low density lipoproteins(LDL-C)(r=-0.42,P＜0.01).Serum levels of apolipoprotein M were correlated negatively with the score of coronary stenosis(r=-0.62,P＜0.01).The results suggest that apolipoprotein M might be related to the development of coronary artery disease.

Objective To investigate the changes of cortical thickness and surface area in patients with bipolar depression(BD),and to explore the relationship between abnormal changes in gray matter and clinical symptoms.Methods 28 BD patients and 28 healthy controls underwent T1-weighted MRI.The Freesurfer software was used to process the T1 images,which used a set of automated sequences to analyze cortical thickness and surface area on 66 regions (33 regions of each hemisphere),and the correlation with clinical features was also calculated.Results Compared with controls,BD patients showed thinner cortical thickness in left medial orbitofrontal cortex((2.40±0.12) mm vs (2.55 ±0.18) mm,P=1.2× 10-3) and left rostral anterior cingulate((2.66±0.21) mm vs (2.88±0.27) mm,P=3.1 × 10-4),and smaller area of left cuneus((1 443.13± 131.00) mm2vs (6 634.70±600.16) mm2,P=2.7× 10-4) and right superior frontal gyrus ((6 634.70±600.16) mm2vs (7 300.50±653.39) mm2,P=1.3× 10-3).In addition,the negatively correlation was found between the cortical area of left cuneus and effective illness duration (r=-0.471,P=0.018),and the cortical thickness in left rostral anterior cingulate and total score of HAMD-17(17-item Hamilton Rating Scale for Depression) (r=-0.508,P=0.009).Conclusion There are abnormal altertion of cortical thickness and cortical areas of emotional circuit in bipolar depression,but the brain areas are not completely overlapping.Correlation analysis suggests that cortical thickness and area is related to different clinical features.

Objective To compare the clinical effect of preserving the ascending uterine artery in laparoscopic hysterectomy with traditional laparoscopic hysterectomy.Methods 112 patients with various kinds of uterine benign lesions performed by hysterectomy were randomly divided into the preserving the ascending uterine artery in laparoscopic hysterectomy group(observation group,n =60) and traditional laparoscopic hysterectomy group (control group,n =52),The operation time,the intra operative blood loss,the postoperative complications rate and the perimenopausal symptoms in two groups were compared as well as the levels of blood calcium and the levels of hormone before operation,the first month,the sixth month,the first year,and the third year after operation.Results The opreration time [(95.5 ± 22.8) win] and intraoperative blood soss [(81.3 ± 11.6) ml] of the observation group was more than those of the control group [(68.8 ± 7.2) ml] (t =3.772,4.418,all P ＜ 0.05).The postoperative abnormal uniation and defection (1.7％) and fever incidence (1.7％) of the observation group was lower than those of the control group (11.5 ％,13.5 ％) (x2 =4.623,5.843,all P ＜ 0.05).The estradiol level of the observation group began to fall in 6 months postoperative,and had significant differences compare with those of the control group 6 months,1 year,3 years after operation [(328.4 ± 13.4) pmol/L vs (304.8 ± 15.2) pmol/L,(311.3 ± 12.9) pmol/L vs (248.5 ±13.6)pmol/L,(258.3 ± 11.4)pmol/L vs (138.7 ± 12.7)pmol/L] (t =8.758,25.121,52.688,all P ＜0.05).There was significant difference in the level of follicle stimulating hormone (FSH) and luteinizing hormone (LH) between the two groups 1,3 years after operation(t =-90.540,-123.229,-14.757,-19.845,all P ＜ 0.01).There was significant difference in the level of calcium between the two groups 1,3 years after operation(t =3.261,3.050,all P ＜0.05).The perimenopausal symptoms of the observation group was less than those of the control group(x2 =6.142,3.471,4.976,6.646,4.633,all P ＜ 0.05).Conclusion Laparoscopic hysterectomy of preserving the ascending uterine artery is safe and reliable;the recently ovarian function of the patients with hysterectomy is to maximize the preservation in the treatment,the women's quality of life will be to improved.

BACKGROUNDTo study the expression of Skp2 and c-myc in non-small cell lung cancer (NSCLC) and to investigate their relationship and clinical significance.

METHODSThe expression of Skp2 and c-myc was detected in 42 NSCLC, 10 pulmonary benign disease and 8 epithelial dysplasia tissues by immuno-histochemistry.

RESULTSThe positive rate of Skp2 stain was 24.83%±13.64% in NSCLC tissues, which was significantly higher than that in pulmonary benign disease tissues (3.07%±1.32%)(P < 0.001) and that in dysplasia tissues (13.89%±3.95%)(P < 0.05). The expression level of Skp2 was closely related to cell differentiation (P < 0.001), TNM stages (P < 0.01) and lymph node metastasis (P < 0.05), but not to pathological type of NSCLC. There was a positive correlation between Skp2 and c-myc expression (r=0.448, P=0.003). The concurrent high expression rate of Skp2 and c-myc was 38.1% (16/42), which was closely rela- ted to TNM stage (P < 0.05), but not to cell differentiation, pathological type or lymph node metastasis.

CONCLUSIONSOverexpression of Skp2 may play important roles in carcinogenesis and development of NSCLC, and it may cooperate with c-myc protein.

Objective To evaluate the early diagnostic value of circulating microRNA-1 (miR-1) on acute myocardial infarction (AMI). Methods A prospective cohort study was conducted. The patients with chest pain admitted to the Second People's Hospital of Wuxi from November 2012 to June 2015 were enrolled. According to AMI diagnostic criteria, the patients were divided into AMI group and non-AMI group, and healthy individuals during the same period were served as heath controls. The venous samples of the onset patients were collected within 3 hours after admission. The plasma miR-1 was determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR), and the levels of plasma cardiac troponin I (cTnI) and MB isoenzyme of creatine kinase (CK-MB) were measured by electrochemiluminescence. The correlation between plasma miR-1 and cTnI as well as CK-MB was performed by Spearman analysis. The early diagnostic performance of plasma miR-1, cTnI, and CK-MB for AMI was estimated by receiver operating characteristic (ROC) curve analysis. Results There were 127 patients in AMI group, and 107 in non-AMI group, including 82 patients with angina pectoris, 2 with pulmonary embolism, 3 with aortic dissection, 2 with acute pericarditis, 3 with myocarditis, 13 with acute heart failure, and 2 with peptic ulcer. Ninety volunteers were served as healthy controls. There was no difference in clinical characteristics including gender and hyperlipidemia between AMI group and non-AMI group. The expressions of plasma miR-1, cTnI and CK-MB were significantly increased in AMI patients as compared with those of the healthy controls [miR-1 (2-ΔΔCt): 4.32±2.60 vs. 1.44±0.75 and 0.98±0.18, cTnI (μg/L): 3.23 (0.63, 10.70) vs. 0.02 (0.00, 0.17) and 0.00 (0.00, 0.00), CK-MB (U/L): 32.40 (14.20, 95.40) vs. 14.40 (11.20, 17.10) and 8.90 (8.28, 9.50), all P < 0.01]. The expression of plasma miR-1 had a significantly positive correlation with cTnI and CK-MB in AMI patients (r1 = 0.395, r2 = 0.490, both P < 0.000). It was demonstrated by ROC curve analysis that the area under ROC curve (AUC) for the diagnostic value of miR-1 on AMI was 0.905 [95% confidence interval (95%CI) = 0.860-0.950, P = 0.000], the sensitivity was 86.6%, and the specificity was 95.4%; the AUC for cTnI was 0.908 (95%CI = 0.870-0.946, P = 0.000), the sensitivity was 81.9%, and the specificity was 95.9%; the AUC for CK-MB was 0.795 (95%CI = 0.736-0.854, P = 0.000), the sensitivity was 63.0%, and the specificity was 92.9%. Conclusions Plasma miR-1 has the capacity in early diagnosis of AMI, superior to CK-MB, and equal to cTnI. It can provide additional diagnostic information beyond cTnI. The diagnostic accuracy for early AMI can be improved with the combination of plasma miR-1 and cTnI.

The objective of this experimental study was to assess the effects of a new kind of drug eluting coronary stent. Fourteen mini-pigs were used; seven normal stainless stents and seven new drug eluting stents were implanted in their normal coronary arteries, respectively. Angiography was performed and followed by pressure-fixation of the coronary arteries for light and electron microscopic examinations at the end of three months after implantation. Repeated angiography showed that all the stented coronary segments were open. With no additional antithrombotic treatment, there was no thrombus formed in the stented coronary segments. Scanning electron microscopy analysis showed the implanted stent surface was covered by endomembrane without thrombus formation. The endothelial cell in the membrane was clear and lined by the direction of blood flow. Histomorphological analysis revealed the neointima in normal stainless stent group was thicker than that in new drug eluting stent group, and the neointima was composed of smooth muscle cell and extracellular matrix. The result of this study shows that this kind of stent could reduce the rate of the re-stenosis and occlusion of PTCA. This stent can be used in clinical trials.
Angioplasty, Balloon, Coronary ; instrumentation ; Animals ; Coated Materials, Biocompatible ; chemistry ; Coronary Restenosis ; prevention & control ; Drug-Eluting Stents ; Random Allocation ; Swine ; Swine, Miniature ; Titanium ; chemistry

Objective To examine the benefits of ECMO for potential organ donors with hemodynamic instability after brain death.Methods Three brain-dead potential donors who presented with hemodynamic instability despite maximal medical management,finished a declaration of brain death,that were supported by extracorporeal circulation membrane oxygenation (ECMO).Results Donor organs,including six kidneys,and two livers,were harvested from the three donors under ECMO support,leading to 8 successful transplantations.The organs functioned well and the recipients made full recoveries.Conclusion Our experience indicates that ECMO allows for the maintenance of abdominal organ tissue perfusion without warm ischemia before organ procurement,providing sufficient time for safe organ donation procedures and reducing the risk of unpredictable cardiac arrest that could result in the donor death and graft loss.