Objective To evaluate the effects of high glucose on autophagy and apoptosis of podocyte and explore the signaling pathway in high glucose-induce podocyte autophagy.Methods Differentiated mouse podocytes were exposed to high glucose(30 mmol/L) or rapamycin (autophagy enhancer,1 μg/L) or LY294002 (a selective PI3K inhibitor,50 mmol/L) for 24 h.The formations of autophagy were observed by electron microscopy and acridine orange staining.Apoptosis was evaluated by flow cytometry.The expression of autophagy protein LC3-Ⅱ/Ⅰ and Beclin-1 as well as the phosphorylation of AKT and mTOR were examined by Western blotting analysis.Results High glucose induced podocytes apoptosis,increased autophagy and the expression of autophagy-associated proteins (all P ＜ 0.05).Rapamycin further increased the expression of LC3-Ⅱ and Beclin-1 protein (all P ＜ 0.05),but LY294002 inhibited partialiy the protein expression of LC3-Ⅱ and Beclin-1 induced by high glucose (both P ＜ 0.05).Treatment with rapamycin increased the phosphorylation of AKT,but reduced that of mTOR in podocytes.Moreover,LY294002 inhibited phosphorylation of both AKT and mTOR (both P ＜ 0.05).Conclusions High glucose promotes podocyte autophagy and apoptosis.High glucose-induced autophagy is mediated partly through PI3K-AKT-mTOR signaling pathway.

Background The measurement of retinal nerve fiber layer (RNFL) by Cirrus optical coherence tomography (OCT) has been applied widely in ophthalmology.However,previous assessment of RNFL is based on spatial measurement,while the evaluation based on optical characteristics of OCT can offset the shortcomings of spatial measurement in some eye diseases.Objective This study was to analyze the optical characters of RNFL measured by OCT and its correlation with age in healthy Chinese individuals.Methods Four hundred and six normal healthy subjects were collected in Shanghai Shibei Hospital from June 2011 to June 2012,with the age of 40-83 years old.Macular RNFL was unilateral imaged using a Cirrus OCT device with 5 line raster macular scan mode under the approval of Ethic Committee of Shanghai Shibei Hospital and informed consent of subjects.A customized software was used to measure RNFL optical characters,including absorbance (A value) and attenuate coefficient.The difference of the measured parameters in different genders was compared.The correlations between RNFL optical characters and age were analyzed by linear regression analysis.Results The data of 353 eyes were included in the final outcomes.The RNFL thickness,A value and attenuate coefficient were (35.1 ±4.4) μm,121.6 ±5.3 and 2.06 ±0.25,respectively.RNFL thickness and A value showed significantly negative correlations with age (r=-0.487,-0.571,both at P＜0.01) with the regression equation Y=-0.17 X+45.23 and Y=-0.24 X+ 135.82,while a positive correlation was found between the attenuate coefficient and age (r=0.368,P＜0.01) with the regression equation Y=0.01 X+1.63.RNFL thickness and A value showed weaker positive correlations with OCT signal intensity (rs =0.128,P =0.016;rs =0.284,P＜0.01),but no remarked correlation was seen between the attenuate coefficient and OCT signal intensity (rs =-0.053,P=0.319).Conclusions Age of subjects affects the parameters of OCT optical characteristics in normal populaition.Age-related RNFL change in optical characters of OCT should be considered before concluding optical parameters.

Objective To evaluate the effects of autophagy on oxidative stress induced by contrast media in podocytes.Methods The differentiated mouse podocytes were exposed to contrast media (Iopromide,50 mg/L)、rapamycin (Rap,autophagy enhancer,1 ng/L),3-methyladenine (3-MA,autophagy inhibitor,2 mmol/L) for 2 hours.The expression of autophagy protein LC3-Ⅱ and Beclin-1 as well as oxidative stress-related proteins Catalase,MnSOD were detected by Western blot.The formations of autophagy were observed by MDC staining,and the levels of reactive oxygen species (ROS) by CM-H2DCFDA staining.Cell activity was evaluated by CCK8 assay.Results Both the levels of oxidative stress and autophagy in podocytes increased when stimulated by contrast media,the expression of LC3-Ⅱ and Beclin-1 were enhanced,Catalase and MnSOD were inhibited (all P ＜ 0.05).Rapamycin increased the expression of Catalase,MnSOD and cell activity of podocytes,reduced the generation of ROS (all P ＜ 0.05),but in Rap group,cell activity showed no significant difference (P ＞ 0.05).3-MA decreased the expression of Catalase 、MnSOD and inhibited the cell activity of podocyte,increased the generation of ROS (all P ＜ 0.05).Conclusion Autophagy protects podocyte from contrast media by the means of reducing oxidative stress.

Objective To analyze the pathogenesis and prevention and treatment of limb deep venous thrombosis complicated by postoperative gynaecological malignant tumours.Methods A retrospective analysis of 22 cases with limb deep venous thrombosis complicated by postoperative gynaecological malignant tumours in our hospital were analyzed.According to the situation of patients,surgery or conservative treatment was conducted.The clinical efficacy after treatment was observed.Results In 22 cases,6 cases underwent surgical treatment,16 cases underwent conservative treatment.After treatment,18 cases were cured,3 cases were effective,1 case ineffective.After treatment,the TT,Pt,APTT and FBG levels were (24.17 ±2.13)s,(18.25 ±2.13)s,(31.68 ± 10.30)s and (2.26 ±0.52) g/L,which were all better than before treatment,the differences were statistically significant (t =10.362,9.774,12.281,8.462,P =0.028,0.032,0.012,0.029).Followed up for 3 months,no recurrence of LDVT was observed.Conclusion The cases of gynecological malignant tumor have higher risk of limb deep vein thrombosis,we should taken prevention actively,once diagnosed should take effective intervention measures to improve the prognosis.

Objective To investigate the protective effect of Ganyu capsule on the experimental hepatic injury in mice and rats.Methods Acute hepatic injury was induced by intraperitoneal injection of 0.1 %CCl4 10mL/kg and D-galactosamine 500 mg/kg in mice;Cirrhosis was induced by 40 %CCl4 adding with variousagents in rats.The biochemical parameters such as serum ALT were examined and the histopathological changes of hepatic tissue was measured.Results Ganyu capsule could obviously inhibit the increase of serum ALT and AST activity and reduce the content of collagen in liver and the deseverity of hepatic fibrosis.Conclusion Ganyu capsule has protective effects on the acute and chronic hepatic injury in mice and rats.

No abstract available.
Cervical Intraepithelial Neoplasia

ObjectiveTo obtain stable animal models and observe Helicobacter hepaticas (Hh)colonization and pathological claracteristics,through infecting different mice strains with Hh.Methods SPF-class male BABL/c Cr,SCID/Cr and C57BL/6 Cr mice were inoculated 0.2 ml Hh standard strain ATCC51450 bacterial suspension (1 × 108CUF/ml),inoculated for 3 times with 48 hours intervals,the control group was fed with the same volume of PBS.Mice were executed at 4 weeks,8weeks and 16 weeks since last Hh inoculation,and mice esophagus,stomach,jejunum,ileum,cecum,colon,liver and pancreas tissue were taken for histopathology examination,Micro-aerobic bacteria isolation,culture and identification and Hh specific 16S rRNA gene amplification.Results The colonization rates of Hh in cecum after inoculated in BALB/c Cr mice and SCID/Cr mice at 4 weeks,8 weeks and 16 weeks were all 8/8,colonization rates in colon at 4 weeks,8 weeks and 16 weeks were 4/8,5/8,5/8 and 3/8,6/8,5/8 respectively,colonization rates in ileum and jejunum at 16 weeks were 1/8,colonization rates in liver at 8 weeks and 16 weeks were 2/8,3/8 and 2/8,2/8respectively.The colonization rates of Hh in cecum after inoculated in C57BL/6 Cr mice at 4 weeks,8weeks and 16 weeks were 1/8,2/8 and 2/8 respectively,colonization rates in colon at 8 weeks and 16weeks were 1/8,2/8 respectively.Compared with C57BL/6 Cr mice,the inflammatory changes in liver,cecum and colon were more significant in Hh infected BALB/c Cr and SCID/Cr mice (P＜0.01),and histological scores gradually increased as infection time extended (P＜0.05,P＜ 0.01 ).The histological scores were significantly higher in those with colon and liver Hh bacterial colonization than those without Hh bacterial colonization (P＜0.05).The histopathological score of cecal tissue was positively correlated with the density of Hh colonization.ConclusionDifferent mice strains are with different susceptibility to Hh,and better Hh infection model can be obtained in Hh inoculated BALB/c Cr and SCID/Cr mice.

INTRODUCTIONHelicobacter (H.) hepaticus infection causes chronic active hepatitis and induces hepatocellular tumours in A/JCr mice, but evidence of this in humans is scarce. This study aimed to demonstrate the correlation between H. hepaticus and human primary hepatocellular carcinoma (HCC).

METHODSThe sera of 50 patients with primary HCC were tested for the presence of anti-H. pylori and anti-H. hepaticus immunoglobulin G (IgG) antibodies. The liver tissues of patients who tested positive for serum antibody were analysed for H. hepaticus-specific 16S rRNA, H. hepaticus cdtB, H. pylori cagA, H. pylori vacA and H. pylori ureC genes using polymerase chain reaction.

RESULTSAfter the anti-H. pylori antibodies in the serum samples were absorbed by H. pylori antigen, the anti-H. hepaticus IgG serum antibody detection rate was 50.0% in patients with primary HCC. This was significantly higher (p < 0.001) than the detection rate in the benign liver tumour (7.7%) and normal liver tissue (6.3%) groups. Of the 25 primary HCC samples that tested positive for anti-H. hepaticus IgG serum antibody, the H. hepaticus-specific 16S rRNA gene was detected in nine (36.0%) samples. Sequencing showed that the polymerase chain reaction-amplified product exhibited 95.5%-100% homology to the H. hepaticus-specific 16S rRNA gene. Among these nine primary HCC tissue samples, the H. hepaticus cdtB gene was detected in four (44.4%) samples, while no such expression was observed in the benign liver tumour or normal liver tissue groups.

CONCLUSIONThe present study identified the presence of H. hepaticus infection in patients with primary HCC using serological and molecular biological detection, suggesting that H. hepaticus infection may be involved in the progression of HCC.

Adult ; Aged ; Carcinoma, Hepatocellular ; microbiology ; DNA, Bacterial ; genetics ; Female ; Helicobacter Infections ; genetics ; microbiology ; Helicobacter hepaticus ; genetics ; isolation & purification ; Helicobacter pylori ; genetics ; isolation & purification ; Humans ; Immunoglobulin G ; blood ; Liver Neoplasms ; microbiology ; Male ; Middle Aged ; Polymerase Chain Reaction

Osteoarthritis (OA) is a common degenerative disease. Its most significant pathological change is destruction of articular cartilage and the main clinical symptoms are pain and dysfunction of joints. Recent studies have shown that the expression of non-coding RNA (ncRNA) in chondrocytes can abnormally up-regulate or down-regulate and alter the activities of chondrocytes like their proliferation, migration and apoptosis, thus leading to the occurrence and development of osteoarthritis. Exosomes are extracellular vesicles with a diameter of 40-100 nm, which are secreted in intercellular fluid, act as medium of intercellular communication. They protect ncRNA, protein, lipid and other bioactive materials from enzymatic degradation by encapsulating them and transferring to sibling chondrocytes, due to their good tissue permeability. They can also improve communication between cells and regulate the activities of chondrocytes. Thus, exosomes behave like gene carriers. The ncRNA carried by exosomes can supplement or adsorb the abnormal ncRNA in chondrocytes, so as to regulate the activity of chondrocytes, and is therefore considered as a possible candidate with capabilities to repair cartilages. In this study we reviewed existing literatures related to the roles and effects of exosome miRNA, lncRNA and circRNA on osteoarthritis. We also reviewed the pathogenesis of exosome ncRNA in osteoarthritis.