Objective:To explore the effect of leptin on proliferation of human hepatocellular carcinoma cells Huh 7 and its probable molecular mechanism.Methods:The human hepatocellular carcinoma cells Huh 7 cultured in vitro was treated with leptin at different concentrations.The proliferation of the cells was measured by MTT assay.The cell cycle was monitored by flow cytometry analysis(FCM).The expression of cell cycle regulatory proteins Cyclin D1 and P21waf1 were determined by immunocytochemistry and imageanalysis system,and real-time quantitative PCR.Results:Leptin significantly raised the proliferation rate of Huh 7 cells.The effect was dose and time-depended partly.Leptin promoted Huh 7 cells in entering the S phase from the G0/G1 phase.mRNA of Cyclin D1 in the leptin treated Huh 7 cells was increased,as compared with that in the control cells(P

Objective: To observe the effect of leptin on proliferation and invasion of human hepatocellular carcinoma cell line Huh-7 and explore its possible molecular mechanism. Methods: Huh-7 cells were cultured and treated with leptin at different concentrations of 5, 10, 50, 100 and 200 ng/mL for 12, 24 and 48 h. Huh-7 cells without leptin treatment were used as control group. The proliferation rate of the Huh-7 cells was measured by MTT assay. The change of cell cycle was detected by flow cytometry analysis. Transwell chamber assay was performed to determine the effect of leptin 100 ng/mL on the invasion capability of Huh-7 cells 24 h later. The effect of leptin on mRNA expression of matrix metalloproteinase (MMP) of Huh-7 cells was detected by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). Results: Leptin significantly promoted the proliferation of Huh-7 cells. The effect was dose-dependent in certain degree. Flow cytometry analysis found that leptin significantly induced decrease in proportion of cells in G0/1 phase and significantly increased cell proportion in S phase. The invasion capability of Huh-7 cells was significantly increased after leptin 10 ng/mL treatment for 24 h (P <0.01). The number of cells penetrated polycarbonate membrane were (41.80 ± 3.19) and (18.20 ± 2.86), respectively. Real time fluorogentic quantitative PCR analysis revealed that the expression level of MMP-2 and MMP-9 mRNA expression were 4.25-fold and 2.73-fold as the control group (P <0.01), respectively. Conclusion: Leptin significantly increases the proliferation and invasion capabilities of Huh-7 cells in vitro. The possible mechanism may be related with up-regulation of MMPs level.

[Objective]To explore the effect of combined therapy of Chinese and western medicine on Bell paralysis.[Methods] Randomly divide 62 cases of Bell paralysis into 2 groups who were treated respectively with combined therapy of Chinese and western medicine and western medicine.[Results] For effective rate,the treatment group(91.1%) was markedly higher than control group(67.8%),there was obvious difference between the 2 groups.[Conclusions] Combined therapy of Chinese and western medicine has marked therapeutic effect on Bell paralysis.

Objective To investigate the recurrence and influence factors of spontaneous pneumothorax in patients after thoracoscope surgery. Methods From March 2009 to March 2011,218 patients with spontaneous pneumothorax after thoracoscope surgery in our hospital were surveyed by the self-made general data questionnaire. The clinical data were analyzed by descriptive statistics and binary classification logistic regression analysis of SPSS 19. 0 to find out the infection influential factors of spontaneous pneumothorax for patients after thoraco-scope surgery in order to provide reference basis for reducing postoperative recurrence. Results The recurrence rate of spontaneous pneumo-thorax was 5. 05%(11/218). Multiple logistic regression analysis showed that recurrent pneumothorax ( OR=2. 432,P=0. 000) and multi-ple pulmonary bulla (OR=1. 918,P=0. 000) were the risk factors of spontaneous pneumothorax for patients after thoracoscope surgery. Conclusion The recurrence rate of spontaneous pneumothorax is high,we should strengthen the management of high-risk population and im-prove the prognosis of patients.

The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.

Objective To investigate the characteristics and plausible mechanisms of attention network impairments in hyperthyroid patients.Methods 34 hypertyroid patients and 33 normal controls performed the attention network test (ANT),which was used for assessing three components of attentional networks including alerting,orienting,and executive control.Results The mean scores for alerting and orienting networks were significantly shorter (P ＜ 0.01) in hyperthyroid patients (respectively,(20.00 ± 22.47) ms,(35.26 ± 22.65) ms)than those in normal controls (respectively,(35.70 ± 17.62) ms,(50.61 ± 23.66) ms).The mean reaction time for executive control network was significantly longer(P ＜ 0.01) in hyperthyroid patients((116.71 ± 40.36)ms)than those in controls ((93.00 ± 29.34)ms).Triiodothyronine(T3) and thyroxine(T4) were positively correlated with the scores for executive control network (respectively,r=0.350,P＜ 0.05; r=0.417,P＜ 0.05) in hyperthyroid patients,whereas no relationship was found between thyroid stimulating hormone(TSH) and the value of executive control network.T3,T4 and TSH were not correlated with the value of alerting and orienting network in hyperthyroid patients.Conclusion Hypertyroid patients have extensive impairments in various kinds of attentional network,rather than a specific deficit.The recession of executive control network accompanies with enhancement of serum levels of T3,T4 in hyperthyroid patients.

In the reform of human resource system at public institutions,public hospitals are challenged with changing employment mechanism and effective mobilization of all-staff's incentives.This paper introduced the classified staff management by Sir Run Run Shaw Hospital,College of Medicine,Zhejiang University,which covered the background,specific methods,purposes,initial results,as well as the key links and problems encountered.The study proved that classified management of the hospital staff helps create a fair and impartial workplace,conducive for sustainable development of the hospital.

Objective To study the roles of TLR2 and TLR4 in the progress of invasive pulmonary aspergillosis(IPA) in experimental mice.Methods The mice were divided into three groups including the group of normal mice,the group of normal mice infected with A.fumigatus and the group of IPA mice.The mice were sacrificed at four time points(8 h,24 h,48 h and 72 h) after infection.The lung tissues from each group were collected for pathological analysis and RT-PcR for detecting the expression level of,TLR2,TLR4 and β-tublin.The ratio of density value of band of each PCR product on electrophoresis to the density value of β-tublin was used to evaluate the expression level of each gene like TLR2.TLR4 and TNF-α.Re-suits The pathological analysis showed the normal structure and no inflananatory reaction in the lungs in the group of normal mice.The infiltration of inflammatory cells,weak injuries and no germination of spore into hypha in the lungs of normal mice infected with A.fumigatus,and serious injuries like destruction of alveolar structure,bleeding,infiltration of inflammatory cells and germination of spore into hypha in the lungs of IPA mice.The expression level of TLR4 at 8 h,24 h,48 h and TNF-α at 24 h and 48 h were lower in IPA mice than that in healthy mice with infection(P<0.05).Conclusion There was low expression of TLR4 and TNF-α in IPA mice lung tissues.Typical pathological injuries in the lungs and germination of spore into hy-pha in IPA mice were observed by the microscope.

Background The rat model of N-methyl-N-nitrosourea (MNU)-induced photoreceptor cell apoptosis is often used to study retinal degeneration.But the changes in the gene expression patterm in retinal degeneration in rats have not been reported.Objective This study was undertaken to investigate regulation of gene expression in the retina of MNU-induced retinal degeneration in rats by performing microarray analysis of retinal RNA.Methods Fifty 6-week-old SD rats were numbered and randomized into the normal group and the model group.The retinal degeneration model was established by a single hypodermic injection of 40 mg/kg of MNU,and the rats in the normal group received equivalent volume of physiological saline in the same way.The rats were sacrificed 12 hours or 24 hours after injection.Retinal sections from the right eyes were prepared for the measurement of the retinal thickness by histopathological examination,and retinas from the left eyes were used to confirm the differential gene expression as detected by microarray (normal group and 12 hours model group).Genes exhibiting changes in expression by ≥2.0 folds were further confirmed using real-time PCR.Results The whole thickness of the retina declined in the rats from the 24 hours model group compared to the normal group and 12 hours model group (t =9.926,P=0.002;t=2.736,P=0.028).The thickness of the outer nuclear layer was (26.58±2.90) μm in the 24 hours model group,showing a significant decrease in comparison with (38.11 ± 1.01) μm in the normal group and (35.07t3.03) μm in the 12 hours model group (t=6.028,P=0.009;t=6.839,P=0.006).However,there was no significant difference in retinal thickness between the normal group and the 12 hours model group (whole thickness:t=1.541,P=0.324;outer nuclear layer thickness:t=2.040,P=0.134).Microarray analysis of the rat genes showed that out of 17 000 genes,142 genes involved in biological process and 94 genes involved in molecular functions were differentially expressed,where most of them participate in the mitogen activated protein kinase signaling pathway,Tolllike receptor signaling pathway and apoptosis pathway.Real-time PCR analysis demonstrated that the expression of CCL2,IL-1b,CCL3,c-fos,c-myc,p53 and MMP3 were consistently up-regulated,conforming with the results from microarray analysis.Conclusions The changes in gene expression pattern appear in the early stage of MNUinduced retinal degeneration.These microarray results provided clues to understanding the molecular pathways underlying photoreceptor degeneration and indicating the directions for future studies.

Objective To evaluate surgical treatment of infected femoral artery pseudoaneurysm.Methods The data on surgical treatment of 45 patients with infected femoral artery pseudoaneurysm admitted from January 2003 to June 2008 were analyzed retrospectively.Fourty-three patients underwent operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis.Two patients were unavoidable to undergo removing of infected femoral artery pseudoaneurysm and ligating the proximal and distal artery of pseudoaneurysm because of severe infection and large volume.Results The patients were followed up from 3 to 12 months(mean 7.82 months).The limbs of all the patients underwent bypass graft with vascular prosthesis were salvaged successfully,patients of which had secondary wound healing and had not intermittent lameness.One of two patients performed ligation of artery was salvaged successfully but had severe intermittent lameness,another patient underwent high amputation above knee because of ischemic gangrene.ConclusionFor infected femoral artery pseudoaneurysm,the operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis is effective and safe.