Adolescent ; Adult ; Diabetes Mellitus, Type 1 ; blood ; complications ; genetics ; Diabetic Angiopathies ; blood ; complications ; Female ; Gene Frequency ; Genotype ; Homocysteine ; blood ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Mutation ; Polymorphism, Genetic ; Regression Analysis

AIM:To observe the expression of connective tissue growth factor(CTGF)in unilateral ureteral obstruction(UUO)rats,and to explore its pathogenic role in renal tubulointerstitial fibrosis.METHODS:48 Wistar rats were randomly divided into sham-operated and UUO group.The rats were sacrificed at day 1,3,7,and 14.The degree of tubulointerstitial damage was scored according to the Masson staining.The mRNA and protein levels of CTGF,transforming growth factor-?1(TGF-?1),collagen Ⅰ(Col Ⅰ),and plasminogen activator inhibitor-1(PAI-1)were detected by reverse transcriptional-polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively.Expression of CTGF protein in the kidney was also assessed using Western blotting.RESULTS:TGF-?1 mRNA level began to increase as early as 1 day after UUO.This increase was followed by the elevation of CTGF mRNA level,which began to increase at third days after UUO(P

ObjectiveTo study the level of temperament and self-esteem of children aged 8～12 in Beijing city,as well as their interaction and influenced factor.Methods203 school children aged 8～12 were assessed with Carey's temperament questionnaire(8～12:MCTQ) and The Self-Esteem Scale(SES).ResultsTemperament and self-esteem of school children aged 8～12 were associated with the parental behavior,especially with work style of their fathers.The rhythmicity was found to be negative correlated to the self-esteem scores,and the intensity of reaction was found to be positive correlated to the self-esteem scores.ConclusionParents' behavior and family environment were important to the temperament and self-esteem of school children.

AAV-ITR single strand DNA mini vector (AAV-ITR ssDNA mini vector) is a novel gene expression vector based on AAV-ITR. We have shown efficient gene expression of AAV-ITR ssDNA mini vector in HEK 293T. Here, we studied the efficacy of gene expression of AAV-ITR ssDNA mini vector in vivo. We injected the skeletal muscle of ICR mice separately with equal molars of AAV-ITR ssDNA mini vector, ITR mutated AAV-ITR single strand DNA mini vector (AAV-ITRmm ssDNA mutant vector), AAV-ITR dsDNA and pUC57-minivector-GFP, combined with TurboFect. Florescence microscope analysis of skeletal muscle section shows that AAV-ITR ssDNA mini vector had higher expression efficiency and longer expression period. We extracted DNA from the muscle three months after injection and quantified three vectors by Real-time PCR. RT-PCR analysis shows that there were highest copy numbers of AAV-ITR ssDNA mini vector existing in muscle. Stable existing of AAV- TR ssDNA mini vector in muscle could be the molecular basis of long term gene expression of the vector. The results suggest that AAV-ITR ssDNA mini vector might be a promising vector for gene therapy.
Animals ; DNA, Single-Stranded ; genetics ; Dependovirus ; genetics ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Mice ; Mice, Inbred ICR ; Muscle, Skeletal ; metabolism ; Real-Time Polymerase Chain Reaction

Objective To study the effects of CD2-associated protein (CD2AP) on podocyte adhesion and extension ability and to explore its possible mechanism. Methods Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium at 33℃permissive conditions. The podocytes were transfected with CD2AP small interfering RNA (siRNA) and serambing sequences labeled with fluorescein were taken as control. The transfected podocytes were trypsinized and seed into collagen IV coated plates. The relative cell adhesion and cell area were examined 90 min later. Apoptotic rates of CD2AP siRNA transfected podoeytes and different PAN concentrations incubated podoeytes were detected by flow cytometer. The distribution of F-actin was observed under laser scanning confoeal microscope. Nephrin protein expression and its phosphorylation level were examined by immunofluorescence and Western blot. Results The relative ceil adhesion of CD2AP siRNA transfected podocytes was apparently lower than that of control group[(41.72±6.07)% vs (64.46±8.53)%, P<0.05]. The cell area analysis had the similar result. The apoptotic rate of CD2AP siRNA transfected podocytes was significantly higher than that of the controls [(5.73±0.61)% vs (3.26±0.45)%, P<0.05]. 100 mg/L PAN could markedly induce podocytes to apoptosis and impair cell adhesion ability (P<0.05). Nevertheless, no significant difference was found in cell body spreading (P>0.05). The distribution of F-actin in CD2AP depletion podocytes was apparently altered. The expression of nephrin protein and its phosphorylation level was conspicuously descended to some degree (P<0.05). Conclusions CD2AP depletion facilitates podocyte apoptosis and impairs cell adhesion function. Cytoskeleton confusion and nephrin signaling weakness caused by CD2AP depletion may he partly responsible for the decline of cell adhesion and spreading.

Objective To explore the relationship between brachial ankle pulse wave velocity (BaPWV) change and carotid artery plaque formation in middle-aged and elderly patients with essential hypertension(EH).Methods A cross sectional study was conducted in 409 patients who were admitted in Hypertension Department in our hospital from January to September 2013.Their age ranged from 35 to 75 years,with mean aged(53.2±15.0) years.Parallel carotid artery ultrasound and BaPWV examination were performed in all patients.According to whether carotid plaques were present,patients were divided into two groups:carotid plaque group and control group.And carotid plaque group was sub-grouped into normal BaPWV group (BaPWV ＜ 1400cm/s) and increased BaPWV group(BaPWV≥1400 cm/s) according to BaPWV levels.The detection rate of carotid plaque was compared between normal BaPWV group and increased BaPWV group.The correlation between BaPWV and carotid plaques was analyzed by multivariate logistic regression analysis.Results Compared with the control group,carotid plaque group showed the prevalence of carotid plaques was increased along with the increases of age(t=11.0,P=11.0),systolic blood pressure (t=3.87,P=3.87),diastolic blood pressure(t=3.70,P=0.00),pulse pressure(t=6.13,P 6.13),total cholesterol levels(t=2.57,P=0.01).The detection rate of carotid plaque was higher in increased BaPWV group than in normal BaPWV group [62.6％(159/254) vs.43.2％(67/155),x2=14.61,P=0.00].After adjusting for age,blood pressure and other cardiovascular risk factors,the multivariate logistic regression analysis showed that increased BaPWV was an independent risk factor for carotid plaques(OR=2.06,P=0.05),and age,smoking,systolic pressure,diastolic pressure,pulse pressure,total cholesterol,low-density lipoprotein cholesterol,triglyceride levels were positively correlated with carotid plaques.Conclusions BaPWV is one of the independent impact factors for carotid plaques,which plays an important role in early diagnosis and screening for subclinical vascular lesions.

0.05).It could also decrease the times of wet manure induced by folia sennae,while it was of no effect on diarrhea induced by castor oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has anti-inflammation,analgesic and anti-diarrhea effect related to the cure for ulcerative colitis.

AIM: To investigate antioxidative and antinitrosative effects of volatile oil from A.longiligulare T.L.Wu on dextran sulfate sodium(DSS)-induced ulcerative colitis mice. METHODS: Balb/c mice were fed with 4% DSS solution for 7 d to induce ulcerative colitis.Using biochemical method,the activity of antioxidative enzyme SOD,MDA and NO were determined in normal,model,SASP and three mouse's groups with low,moderate and high volatile oil from A.longiligulare T.L.Wu respectively.At the same time,the activity of iNOS was also measured by immunohistopathology. RESULTS: The concentration of MDA and NO were reduced and SOD increased significantly in high-and moderate-volatile oil groups compared with those in the model group.The activity of iNOS was reduced significantly in high-and moderate-volatile oil groups compared with those in the model group.The results demonstrated that the expression of iNOS was significantly inhibited in DSSinduced ulcerative colitis mice after being treated with high or moderate-dosage volatile oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has antioxidative and antinitrosative effects which may be one of the mechanism for treating UC.

Objective; To clone, sequence and analyze the coding sequences of the Mr 15000 and Mr 9000 active segments of natural granulysin derived from human CTLs activated by allogenic antigen ,in order to establish the basis for further purifying and investigating the immune - impairing mechanism of granulysin. Methods; The coding sequences of the Mr 15000 and Mr 9000 granulysin gene were amplified from the total RNA of activated CTLs of healthy person after reverse transcription by Nested - PCR, inserted into pET32a ( + ) vectors and then transformed into E. Coli TOP10, respectively. The recons were identified by PCR, endonuclease digestion and sequencing. Results: The whole coding sequences of the Mr 15000 and Mr 9000 active segments were successfully cloned as expected. The accurate pET32a - Mr 15000 and pET32a - Mr 9000 recons were obtained through Colony - PCR, endonuclease digestion and sequencing. There lied polymorphism on the 119 th amino acid of the product encoded by GLS gene. Conclusion; The coding sequences of the Mr 15000 and Mr 9000 active segments of human granulysin can be obtained and cloned by the methods mentioned above and can be used for subsequent research.

The conditions of Se accumulating Agaricus blazei in submerged culture with shake flask were studied in this paper, experimental results showed that the composition of the cultural medium was as follow: corn starch 30g/L,glucose 20g/L,(NH 4) 2SO 4 2g/L, yeast extract 3g/L, KH 2PO 4 2g/L, MgSO 4?7H 2O 1g/L and VB 1 100mg/L,that strains domesticated in slant with 1mg/L Se, 250mL conical bottle contained with 80mL liquid cultural medium, pH 7 , 80r/min and 12 days of cultural period were the optimum cultural conditions,and that on such conditions , dry weight of the mycelium reached 18 6g/L,Se accumulated in the mycelium reached 55 32?g/g and Se accumulation rate reached 5 14%