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.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of C-terminal Src kinase (Csk) in Ang Ⅱ-infused rat model and cultured podocytes,and to explore the role of Csk in Ang Ⅱ-induced cytoskeletal rearrangement of podocytes.Methods Twenty-four Wista rats were randomly subjected to normal saline infusion,or Ang Ⅱ infusion at 400 ng · kg1 · min-1 (via subcutaneous osmotic minipumps) for 2 or 4 weeks.Renal histomorphology was evaluated through electron microscopy.The expression of glomerular Csk was analyzed by immunofluorescence and Western blotting.In vitro,conditionally immortalized mouse podocytes were cultured and treated with Ang Ⅱ doses ranging from 10-9 mol/L to 10-5 mol/L and for different hours.The expression of podocytes Csk was assessed by Western blotting.After transfection to podocytes with Csk siRNA,FITC-conjugated phalloidin was used to stain F-actin,to investigate the role of Csk in Ang Ⅱ-induced or cytochalasin D-induced cytoskeletal rearrangement.Results (1) Examination of Ang Ⅱ infusion rats glomerular and podocyte ultrastructure by electron microscopy revealed foot process effacement and fusion; (2) In Ang Ⅱ infusion rats,the expression of glomerular Csk was increased (P ＜ 0.05); (3) In vitro,Ang Ⅱ-stimuli up-regulated the expression of Csk (P ＜ 0.05),and the effects of Ang Ⅱ were on dose-dependent and time-dependent manner; (4) Ang Ⅱ-induced disruption of F-actin was alleviated by Csk siRNA transfection in cultured podocytes; furthermore,cytochalasin D depolymerized the F-actin cytoskeleton,while Csk siRNA stabilized the actin filaments.Conclusion The enhanced expression of Csk may be involved in Ang II-induced podocytes cytoskeletal rearrangement and foot process fusion.

Objeetive To investigate the clinical efficacy of treating stomach cance belonging to spleen and stomach deficient cold type with self-designed formula and chemotherapy.Methods 100 patients of stomach cancer belonging to stomach and spleen deficient cold were randomly grouped into a control group and a treatment group,with 50 cases in each group.The control group was only treated with chemotherapy,and the treatment group was additionally treated with self-designed Chinese formula on the basis of the control group.After 3 months' treatment,the curative effect,the improvement of life quality,the changes of body weight,the improvement of peripheral blood and immune function,and the occurrence of adverse reaction were observed.The survival rate of patients after 1,3 and 5 years' treatment was investigated.Results ① Short term curative effect:the total effectiveness of the control group and the treatment group was 60.0％ (30/50) and 30.0％ (15/50) respectively,with the treatment group being better than the control group(Z=-2.100,P＜0.05);② life quality:theimprovement rate of life quality in the treatment group and the control group was 64.0％ (32/50) and 20.0％ (10/50) respectively,with the treatment group being better than the control group (Z=-3.259,P＜0.05); ③ body weight:the improvement rate of body weight in the treatment group and the control group was 24.0％ and 16.0％ respectively,and there was no significant difference (Z=-1.815,P＞0.05) ; ④ peripheral blood:the peripheral blood abnormality rates of treatment group was obviously lower than the control group (Z=-4.286,P＜0.05) ;⑤ immune function changes:CD4+、CD8+、and the activity of NK cell of two groups after the treatment [the treatment group was (40.8 ± 4.1)％,(26.1-3.2)％,(83.57-2.52)％ respectively; the control group was (31.7±2.9)％、(33.8±3.0)％、(92.88±3.83)％ respectively] wasobviously improved compared with the same group before the treatment [the treatment group was (34.2±3.5)％,(31.0±2.0)％,(85.01±2.92)％ respectively; the control group was (34.0±3.2)％,(30.9±2.2)％,(85.02± 3.48) ％ respectively].The immune indexes (CD4+、CD8++、CD4+/CDg+and the activity of NK cell)in the treatment group presented significant difference (P＜0.01) than the control group after the treatment.⑥adverse reaction:compared with the control group,the occurrence rate of adverse reaction of the treatment group was obviously lower than the control group,and there was significant difference (Z=-5.297,P＜0.05).⑦survival rate:the survival rate of patients after 1,3 and 5 years' treatment in the treatment group was better than the control group,and there was significant difference (x2=5.263,12.96,7.895,P＜0.05).Conclusion The combination of self-designed Chinese formula and chemotherapy could effectively relieve the clinical symptoms,improve the immunity,reduce the occurrence of adverse reaction,and improve the life quality of patients with stomach cancer belonging to stomach and spleen deficient cold type.

The relationship between visfatin.endothelin, and insulin resistance in the pathogenesis of type 2 diabetes mellitus was explored. Endothelin, visfatin, and other markers were compared. Plasma visfatin was reduced after therapy. Endothelin was positively correlated with fasting insulin, HOMA-IR, HOMA-p, and visfatin. Visfatin was positively correlated with endothelin, HbA1C , fasting insulin, and HOMA-IR. HOMA-IR was an independent related factor in influencing visfatin. The visfatin is related to endothelin and insulin resistance, which may contribute to the pathogenesis of insulin resistance and type 2 diabetes mellitus.

In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P < 0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P < 0.05). No significant difference in the Pin1 expression was found between disease stages (FIGO), pathological grades or pelvic lymph node metastasis status (P > 0.05). The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix (P < 0.05). In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P < 0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.
Cervical Intraepithelial Neoplasia/metabolism ; Ki-67 Antigen/*biosynthesis ; Ki-67 Antigen/genetics ; Peptidylprolyl Isomerase/*biosynthesis ; Peptidylprolyl Isomerase/genetics ; Tumor Markers, Biological ; Uterine Cervical Neoplasms/*metabolism

Aim To observe the effects of rapamycin (Rapa) and starvation-induced autophagy on the morphology of neuronal cells, tau protein aggregation and expression of phosphorylated tau protein, to explore the possible mechanism of cytoprotective effect of these two classical autophagy inducers on phosphorylated tau expressing cells.Methods N2a cells were transfected with GFP-tau plasmid, and equal amount of empty vector was used as control.Then cells were incubated with or without okadaic acid(OA) for 12 h, followed by treatment with autophagy inducers rapamycin(Rapa) and EBSS, autophagy inhibitor Bafilomycin A1(Baf A1) for 6 h.DAB was used to observe tau expression and cell morphology.Confocal microscopy was used to observe the intracellular tau aggregation.TUNEL assay and cleaved caspase-3 level were used to detect cell apoptosis.Immunoblot was used to detect the expression of phosphorylated tau and autophagy-related proteins.Results Our study showed that the N2a cells treated with OA exhibited small cell body, retracted processes and increased tau aggregation, compared with only tau-expressing cells.Rapa and EBSS treatment significantly improved cell morphology, decreased tau aggregation and reduced cell apoptosis.On the contrary, Baf A1 treatment induced aberrant cell shape and increased tau aggregation and cell apoptosis.In addition, Rapa significantly decreased the high molecular weight, phosphorylated tau whereas EBSS especially decreased the low molecular weight phosphorylated tau.Conclusions Rapa and EBSS is alleviate hyperphosphorylated tau-induced cytotoxicity through different mechanism.Rapamycin mainly decreases phosphorylated tau oligomers, while EBSS liable to decrease the soluble phosphorylated tau.

OBJECTIVETo explore the situation and cause of new reported HIV/AIDS cases and death of HIV cases in Henan province from 2008 to 2009.

METHODSThe data of HIV/AIDS cases in Henan province before the end of 2009 was downloaded from "the history card downloading site" of the national online case reporting system on Jan. 5(th), 2010. There were 6990 HIV/AIDS new cases reported from 2008 to 2009, and 1214 cases died. Descriptive study was implemented including population characteristics of new reported HIV/AIDS cases and dead cases from 2008 to 2009. Survival time and cause of death were analyzed.

RESULTSAmong all the dead cases, the Han nationality accounted for 99.0% (1202 cases), and males were more than females (62.0% (753 cases) vs 38.0% (461 cases)), 64.7% (786 cases) were married, those under junior high school education level accounted for 93.7% (1137 cases), AIDS patients accounted for 93.2% (1131 cases) and 6.8% (83 cases) were HIV carriers; 65.5% (795 cases) were former plasma donors and transfusion blood/blood produces. The main cause of death were AIDS-related diseases, the proportion was 71.9% (873 cases). The median survival time after confirmed HIV positive was 62 days (Q(L) = 14 d, Q(U) = 151 d), 35.9% (436 cases) cases died in one month and 79.0% (959 cases) cases died in six months after confirmed HIV positive.

CONCLUSIONThe newly reported dead cases of HIV/AIDS were mainly infected through former plasma donation and transfusion blood/blood produces in 2008 and 2009. The main cause of death was AIDS-related diseases.

Acquired Immunodeficiency Syndrome ; epidemiology ; mortality ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; mortality ; Humans ; Infant ; Male ; Middle Aged ; Young Adult

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation ; methods ; Culture Media ; Pinellia ; physiology ; Protoplasts ; physiology ; Regeneration

SWI1 is a member of a new class of tumor DNA-binding proteins named as the AT-rich interaction domain family (ARID), and considered to bind with AT base pairs specifically. Genomic and functional data support ARID1A as a tumor suppressor because ARID1A/BAF250a (SWI1) subunit of the SWI/SNF chromatin-remodeling complex has emerged as recurrently mutated in a broad array of tumor types. But the crystal structure of SWI1 has not been solved as yet. Using docking and molecular dynamics, we predicted the DNA interaction pattern of human SWI1 ARID and made comparisons with the other two representative ARID family members, human Mrf-2 ARID and Drosophila Dri ARID. Dynamic results revealed that the N-terminal and loop L1 of SWI1 ARID bound with the DNA major groove, while the loop L2 and helix H6 bound with the minor groove. Moreover, it was found that SWI1 ARID bound with DNA apparently in a sequence-nonspecific manner. It was concluded that SWI1 ARID can form stable complex with sequence-nonspecific DNA segment comparing to Mrf-2 ARID/DNA and Dri ARID/DNA sequence-specific complexes.
Binding Sites ; DNA ; chemistry ; metabolism ; DNA-Binding Proteins ; chemistry ; metabolism ; Drosophila Proteins ; chemistry ; Homeodomain Proteins ; chemistry ; Humans ; Models, Molecular ; Molecular Docking Simulation ; Molecular Dynamics Simulation ; Nuclear Proteins ; chemistry ; Protein Structure, Tertiary ; Transcription Factors ; chemistry ; metabolism

Objective To evaluate the role of nutritional parameters in prognosis,especially in the early and late mortality of hospital-acquired acute kidney injury (AKI) patients.Methods This study was a prospective cohort study conducted in a hospital comprising 1500 beds in Shanghai, China. One hundred ninety-four patients with hospital-acquired AKI, as determined using the RIFLE staging criteria,were enrolled as subjects after obtaining informed consent.Patients with AKI caused by postrenal obstruction,glomerulonephritis,interstitial nephritis or vasculitis were excluded.Nutritional evaluation,including subjective global assessment (SGA),anthropometric and laboratory examination,was conducted. Other laboratory measurements and clinical data were recorded.The primary outcome was early mortality (≤ 7 days) and late mortality (8-28 days) after enrolling into the study. Results AKI patients at enrollment were characterized by a high prevalence of malnutrition as determined by SGA, anthropometric and laboratory examination.Univariate analysis showed that the SGA,the serum levels of prealbumin,cholesterol and total lymphatic cells, and the Maastricht index were significantly different among early mortality,late mortality and survival groups.The serum prealbumin and cholesterol levels in the early death group were significantly lower than those in the survival and late death groups (P＜0.05).Multivariate analysis revealed that SGA,albumin,prealbumin and cholesterol remained independently and significantly associated with early mortality after adjusting for age,sex,dialysis,ventilation,hemoglobin,platelets,bilirubin,and Glasgow coma score.The areas under the receiver operating characteristic curve to predict early mortality for albumin,prealbumin and cholesterol were 0.591,0.736 and 0.603,respectively,with that of prealbumin significantly higher than others (P＜0.05). Conclusion Low levels of serum prealbumin,albumin and cholesterol at enrollment are independtly associated with increased early mortality in hospital-acquired AKI patients.