Objective To study the effects of salt sensitivity on evolution of blood pressure and develope- ment to hypertension from adolescents to youth.Methods A baseline survey was carried out in 4623 adolescents aged 6-15 years old in Hanzhong rural area in 1987,310 of them were recruited for determination of salt sensitiv- ity using the tests of oral saline load and furosemide sodium-volume depletion.Salt sensitivity (SS) were diag- nosed in 101 while 209 subjects as no-sah sensitivity (NSS).This cohort of adolescents were followed up for av- erage 18 years.Results The response rate for this cohort of adolescents was 71.9%.At the end of follow up period,BP in subjects with baseline SS was higher in youth than that in NSS (SBP:122.9?13.1 vs 117.3?12.4, P

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

Objective To investigate the expression and function of integrin-iinked kinase (ILK) in the transdifferentiation process of primary tubular epithelial cell( TEC) and its effect on the accumulation of extracellular matrix (ECM) in the aging process. Methods Primary TEC was cultured from kidneys of male Wistar rats aged 3 and 24 months. Then the primary TEC was stimulated by TGF-? at the concentrations of 0, 5, 10 and 20-40/?g/L for 24 h. The TEC expressions of ?-SMA,ILK and F-actin were detected by immunocytochemistry or indirect immunofluorescence. FN or ILK protein expression levels were tested by Western-blot. DIG-labelled cRNA probe of rat ILK in situ hybridization was obtained by transcription in vitro, with which ILK expression level and the location of TEC were detected. Results ?-SMA expression levels were very low in young or aging rat TEC on the base situation, no significant difference was found between the two groups. However, ILK and F-actin levels in 24 months group were higher than in 3 months group. Along with increasing concentration of TGF-?, ?-SMA, ILK and F-actin levels were increased. The expression of ILK was associated with F-actin expression. FN and ILK expression levels were significantly lower in 3 monthsrats TEC than in 24 monthsrats, P

OBJECTIVETo study the role of Mycobacterium tuberculosis in the pathogenesis of sarcoidosis.

METHODSArchival material of 22 patients with a histologic diagnosis of sarcoidosis were retrieved. Real-time fluorescent polymerase chain reaction (PCR) was used to detect DNA fragments of the complex-specific insertion sequence IS6110 of Mycobacterium tuberculosis in formalin-fixed and paraffin-embedded biopsy samples.

RESULTSAmong the 22 samples studied, Mycobacterium tuberculosis DNA was detected in 11 cases. The sequence of PCR amplified IS6110 DNA fragments completely matched with the related sequence in Mycobacterium tuberculosis gene.

CONCLUSIONSMycobacterium tuberculosis DNA is identified in a certain proportion of patients with a clinicopathologic diagnosis of sarcoidosis. Mycobacterium tuberculosis may be an important etiologic agent, at least in some of these patients.

Adult ; DNA, Bacterial ; analysis ; Female ; Fluorescence ; Follow-Up Studies ; Humans ; Lymph Nodes ; microbiology ; pathology ; Male ; Middle Aged ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Sarcoidosis ; microbiology ; pathology

ObjectiveTo study the quality control methods for 99Tcm-TRODAT-1 kit and injection.MethodsThe appearance,pH,contents of the bases,the labeling yield,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 kits from three different batches were examined.The kit stability was estimated under different conditions.The transparence,pH,radiochemical purity,half-life,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 injection were tested.ResultsThe 99Tcm-TRODAT-1 kit and injection were both achromous and transparent,with pH values being 5.9 ± 0.1 and 5.5 - 7.0 respectively.The contents of stannous chloride and TRODAT-1 were stable.The labeling yield of the kit and the radiochemical purity of the injection were both ≥95％.The asepsis test demonstrated that the characters of 99Tcm-TRODAT-1 kit and injection were qualified.TRODAT-1 kit was stable at 0 -4 ℃ for 6 months or at room temperature (20 -25 ℃ )for 10 days,and the radiochemical purity of the injection was still ＞ 90％ at room temperature for 8 hours.ConclusionsThe quality control methods for 99Tcm-TRODAT-1 kit are simple and practical.The kit and injection are qualified and can be used for clinical application.

OBJECTIVEOccult hepatitis B infection of voluntary blood donors has been plagued in the serum screening. Determined the OBI through the highly sensitive detection methods Nest-PCR among the blood donors, and then learned occult HBV infection and analysed the genotypes of this area.

METHODS10 080 serums of donors were determined respectively by the imported Abbott HBsAg kit and Beijing Wantai anti-HBc and anti-HBs reagents, obtained the gene and detected DNA sequences by the high sensitive Nest-PCR method.

RESULTSAmong 10 080 cases of unpaid blood donors, 108 cases were detected HBsAg positively by Abbott sensitivity kit (positive rate of 1.07%), 767 cases were anti-HBc single - positive (positive rate of 7.67%). 25 patients screened blood donors who tested negative for serum HBsAg and positive for HBV DNA in the 10 080 cases. Occult HBV infection incidence rate was 0.25%. 12 cases were HBV genotype C (48%), 13 cases were genotype B (52%), and no other genotypes. Genotype B has no statistically significant difference to genotype C (P > 0.05). Sequence analysis showed that 5 patients in the HBsAg epitope "a" (aa124 - aa147) have mutation (20%).

CONCLUSIONThe high proportion of occult hepatitis B infected among voluntary blood donors in our country. Also genotype and mutation was differences in different regions.

Adolescent ; Adult ; Blood Donors ; DNA, Viral ; blood ; Female ; Genotype ; Hepatitis B ; epidemiology ; Hepatitis B virus ; classification ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Sequence Analysis, DNA

To investigate the effects of lipopolysaccharide (LPS) on autophagy in rat hepatic stellate cells (HSC-T6) and involvement of NF-κB pathway in it.Methods 1)HSC-T6 cells were treated with LPS at the concentraction of 0, 0.01, 0.1, 1, and 10 mg/L for 0, 3, 6, 12, 18, and 24 h respectively, Microtubule-associated protein light chain Ⅱ (LC3Ⅱ) and Beclin1 levels were detected by Western blot; 2)HSC-T6 cells were randomized into groups of control group, LPS group, PDTC group, LPS+PDTC group, PMA group and LPS+PMA group, Western blot assay was used to measure the levels of LC 3Ⅱand Beclin1, immunofluoresence was used to measure NF-κB P65 intracellular distribution.The levels of hydroxyproline (Hyp) were determined by the method of colorimetry and the levels of laminin (LN) and hyaluronic acid (HA) were determined by ELISA in the culture supernatants after corresponding processing .Results The levels of LC3Ⅱand Beclin1 were significantly increased after HSC-T6 cells were treated with LPS at the concentraction of 0.1 mg/L for 6 h and the levels of Hyp, LN, and HA in the culture supernatants increased remarkably as well (P<0.05).PDTC pretreatment increased the levels of LC3Ⅱand Beclin1 in the LPS-treated HSC-T6 cells while decreased the levels of Hyp , LN, and HA significantly (P<0.05).PMA pretreatmen decreased the levels of LC 3Ⅱand Beclin1 in the LPS-treated HSC-T6 cells while increased the levels of Hyp, LN, and HA (P<0.05).Conclusions LPS can promote autophagy and activation of NF -κB pathway in HSC-T6 cells.Activation of NF-κB pathway may inhibit the LPS-induced autophagy of HSC-T6 cells.

OBJECTIVETo investigate the effects of Qushuanling Capsule ( QSLC) on thrombus formation and platelet aggregation in rats.

METHODSArteriovenous bypass, venous thrombosis, and middle cerebral artery thrombosis models were used in rats to investigate the anti-thrombotic effects of QSLC, a compound of nine Chinese herbs. The platelet aggregation induced by adenosine diphosphate (ADP), thrombin or arachidonic acid (AA), as well as the contents of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1α (6-keto-PGF1α) in rat plasma and aortic walls, were determined to investigate the possible mechanisms of the anti-thrombotic effects of QSLC.

RESULTSAfter oral administration with QSLC for 7 days, arteriovenous bypass thrombosis was obviously suppressed compared with the model group, venous thrombosis was also obviously suppressed, rat behaviors were obviously improved, and brain infarct size as well as water content were also reduced. The platelet aggregation induced by ADP or thrombin was inhibited by QSLC, but the drug had no effect on AA-induced platelet aggregation and content of TXB(2) and 6-keto-PGF1α in plasma and the aortic wall.

CONCLUSIONThese results suggest that QSLC can be used in the prevention and treatment of thrombotic diseases, and that its mechanism of action may be related to inhibition of platelet aggregation.

6-Ketoprostaglandin F1 alpha ; blood ; Adenosine Diphosphate ; pharmacology ; Animals ; Aorta ; drug effects ; metabolism ; pathology ; Cerebral Infarction ; blood ; drug therapy ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Middle Cerebral Artery ; drug effects ; pathology ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Thrombosis ; drug therapy ; pathology ; Thromboxane B2 ; blood ; Venous Thrombosis ; drug therapy ; pathology

OBJECTIVETo estimate the human immunodeficiency virus (HIV-1) prevalence of injecting drug users (IDUs) in Chongqing city.

METHODSTo apply BED-capture enzyme immunoassay (CEIA) which was based on the principle of HIV-antibody varies as the disease progress, in order to estimate both the HIV incidence and prevalence of IDUs from two IDUs surveillance sites in Chongqing.

RESULTSDuring the research period, 4711 serum samples were tested by ELISA and 130 were HIV-1 positive, confirmed by Western blot. The prevalence of IDUs surveillance site A from 1999 to 2006 were 0.73%, 2.02%, 1.54%, 2.96% and 2.80%, and the incidence rates were 0.57%, 0.93%, 0,1.24% and 1.68% respectively. The prevalence of IDUs surveillance site B appeared to be 4.21%, 9.96%, 8.13%, and the incidence rates were 0.95%, 1.04% and 0.90% respectively, from 2004 to 2006.

CONCLUSIONMany of the IDUs HIV carriers in Chongqing had been infected for long time, and the incidence rates among them were steady, keeping at the same level for 1-2 years. Promotion on intervention for IDUs had produced certain effects but more attention still needs to be paid.

AIDS Serodiagnosis ; methods ; China ; epidemiology ; Drug Users ; HIV Antibodies ; blood ; HIV Infections ; blood ; epidemiology ; HIV Seroprevalence ; Humans ; Immunoenzyme Techniques ; Incidence ; Population Surveillance ; Prevalence ; Substance Abuse, Intravenous ; epidemiology ; virology ; Urban Population ; statistics & numerical data

OBJECTIVETo examine whether the enhanced expression of CD40L cDNA on murine ovarian cancer (OVHM) cells could induce the secretion of Th1 cytokines from dendritic cells (DC).

METHODSOVHM cells were transfected with the full-length mouse CD40L cDNA by lipofectamine 2000 and then G418 resistant cells as positive cells were selected. They were examined for their expression of CD40L with flow cytometry. Bone marrow cells were firstly depleted of erythrocytes, macrophages, T and B cells with PE-conjugated magnetic beads, and then cultured in 10% FCS RPMI 1640 medium supplemented with recombinant mouse GM-CSF and IL-4 for 10 days. PKH67-labeled tumor cells were cultured with DC, and then the stained cells were analyzed for the expression of MHC-I, MHC-II, CD80, CD86, CCR7 in DC with flow cytometry. The expression of p40, p19, p35, p28, EBI3 subunits, IL-18, IFN-gamma, Mig gene in cocultured DC-tumor cells were detected by RT-PCR.

RESULTSThe CD40L cDNA was successfully transfected into OVHM cells. Bone marrow-derived DCs, when cultured with CD40L/OVHM, formed clusters with the tumors and showed an upregulated expression of MHC- I, MHC-II, CD80, CD86, CCR7. Expression of the IL-12, IL-23, IL-27, IL-18, interferon-gamma (IFN-gamma) and Mig (monokine induced by IFN-gamma) genes was induced in the DCs that were cultured with CD40L/OVHM but not with OVHM cells.

CONCLUSIONThese data directly showed that the expression of CD40L on ovarian cancer cells facilitates the interaction between DCs and tumors, enhances the maturation of DCs, induces secretion of Th1 cytokines, especially for IL-12, IL-23 and IL-27, which maybe one of the possible antitumor mechanism for CD40L-transfected ovarian cancer cell line.

Animals ; CD40 Ligand ; genetics ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Cytokines ; secretion ; DNA, Complementary ; genetics ; Dendritic Cells ; cytology ; metabolism ; Female ; Interleukin-12 ; secretion ; Interleukin-23 ; secretion ; Interleukins ; secretion ; Mice ; Ovarian Neoplasms ; metabolism ; pathology ; Th1 Cells ; secretion ; Transfection