Objective To find out severity and types of syphilis infection in blood donors, attendants and persons to have their premrital medical examination in Zhoushan City and offer a new measure for prevention and treatment of syphilis. Methods Totally, 174 589 blood donors, attendants and persons to have their premarital medical examinations were screened preliminarily for inapparent syphilis with non-TPHA, and then TPHA was applied to confirm the diagnosis, according to the National Standard No. GB 15974-1995, combining with clinical symptoms and physical check-up. Results A total of 1 327 cases of syphilis from 174 589 samples tested, including blood donors, attendants and persons to have their premarital medical examinations, were diagnosed, with an inapparent infection rate of 7. 60‰ in average, 6. 42‰in males and 8. 74‰ in females, with a sex ratio of 0.71 (X2 = 29. 92, P

OBJECTIVETo evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide.

METHODSNinety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis.

RESULTSThe levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced.

CONCLUSIONIn the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Animals ; Azoospermia ; chemically induced ; metabolism ; Cyclophosphamide ; toxicity ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; metabolism ; Immunohistochemistry ; Insulin-Like Growth Factor I ; biosynthesis ; Male ; Oligospermia ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism

OBJECTIVETo establish a rapid method for detection of alpha-globin gene αααbased on droplet digital PCR (ddPCR) technique.

METHODSThe differential sequence between the X1 and Y1 box of α1 gene was selected as the amplicon of the target gene with β-actin as the reference gene. The specific primers and TaqMan probes were designed, and then a quantitative method for detecting the copy number was established based on ddPCR technique. The sensitivity and accuracy of the method were evaluated by detecting 28 samples of known genotypes and 60 clinical samples.

RESULTSThe ddPCR-based method accurately identified the genotypes of all the 28 samples with known genotypes and detected 5 cases of αα/αααfrom the 60 clinical samples, and the results were verified by MLPA. The sensitivity and accuracy of this method were both 100% for detecting alpha-globin gene ααα.

CONCLUSIONThis ddPCR-based method for detecting αααtriplet can be applied for population screening and in routine clinical molecular diagnosis with simple operation, rapid analysis and accurate results.

OBJECTIVETo investigate the effect of Astragalus in regulating the imbalance between naive helper T cells (Th1/Th2) cytokines expression in patients with cervical cancer.

METHODSThirty patients with cervical cancer received intravenous dripping with 20 mL of Astragalus Injection (AI, contained extract from 40 g of crude drug) per day for 1 week, peripheral blood sample was collected from patients separately before and after treatment for extract mononuclear cells by density gradient centrifugation. The positive percentages of CD4+ interferon-gamma (IFN-gamma ) cell and CD4+ interleukin-4 (IL-4) cell in total CD4+ cells were measured by flow cytometry; the contents of IFN-gamma, IL-4 in culture supernate were detected with ELISA; and the expressions of T-cell transcription factor T-cells (T-bet) and GATA-binding protein-3 (GATA-3) were determined by RT-PCR. The data were controlled by those get from 10 healthy persons.

RESULTSCD4+ IFN-gamma positive cell percentage, T-bet mRNA expression level and concentration of IFN-gamma in supernate were significantly lower in patients than those in healthy control respectively, while CD4+ IL-4 positive percentage, level of GATA3 mRNA expression and IL-4 concentration in supernate were insignificantly different between the two groups. After AI treatment, the lowered parameters were up-regulated (P < 0.05), and no obvious change was observed in the CD4+ IL-4 positive cell associated parameters.

CONCLUSIONTh1/Th2 cell function imbalance existed in patients with cervical cancer, showing a Th2 predominant reaction mode; AI can regulate the imbalance, offset to Th1, thus to display its anti-tumor effect.

Aged ; Astragalus membranaceus ; chemistry ; Carcinoma, Squamous Cell ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Middle Aged ; Phytotherapy ; Th1-Th2 Balance ; drug effects ; Uterine Cervical Neoplasms ; drug therapy ; immunology

OBJECTIVETo search for chemical constituents with structural diversity from Laurencia tristicha to supply for biological assay.

METHODCompounds were isolated by means of column chromatography over normal phase silica gel and Sephadex LH-20, recrystallization and HPLC. Structures were identified by spectroscopic methods including 1D NMR, IR and MS. Cytotoxicities of the purified compounds were evaluated by MTT method.

RESULTSeven compounds were isolated from L. tristicha. Their structures were elucidated as cholesterol (1), cholesta- 5-en-3beta, 7alpha-diol (2), beta-stigmasterol (3), phytol (4), zeaxanthin (5), 4 -hydroxybenzaldehyde (6), indolyl-3-carbaldehyde (7). In the cytotoxic assay compound 2 was active against human cancer cell lines HCT-8, Bel-7402, BGc-823, A549 and HELA with IC50 values of 1.90, 2.02, 1.99, 6.52 and 1.20 microg x mL(-1), respectively. Compound 4 showed cytotoxicity against HCT-8 and HELA with IC50 value of 3.51 and 2.04 microg x mL(-1), and other compounds were inactive ( IC50 > 10 microg x mL(-1)).

CONCLUSIONCompounds 1-7 were isolated from L. tristicha for the first time. In additon, compounds 2 and 4 were cytotoxic against several human cancer cell lines.

Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; drug effects ; Cholestenes ; chemistry ; isolation & purification ; pharmacology ; Cholesterol ; chemistry ; isolation & purification ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Laurencia ; chemistry ; Phytol ; chemistry ; isolation & purification ; pharmacology

Gene Products, tat ; genetics ; Gene Targeting ; Hepatitis B virus ; genetics ; growth & development ; Humans ; Recombinant Fusion Proteins ; genetics ; Ribonucleases ; genetics ; Transduction, Genetic ; Virus Replication ; drug effects

OBJECTIVETo observe the inhibitory effect of targeted ribonuclease delivered via adenovirus against HBV replication in vitro.

METHODSThe shuttle plasmids pDC316 were constructed on the basis of the previous plasmids pcDNA3.1(-)/TRL, pcDNA3.1(-)/TR, pcDNA3.1(-)/TRmut, pcDNA3.1(-)/HBVc, and pcDNA3.1(-)/hEDN by subcloning the target gene sequences of TRL, TR, HBVc, and hEDN, respectively. HEK 293 cells were cotransfected with the pDC316 plasmids respectively in the presence of the rescue plasmid pBHGlox(delta)E1,3Cre to yield the recombinant adenoviral vectors which comprised the above genes. After transfection of HepG2.2.15 cells with the vectors, RAd/TRL expression was detected by indirect immunofluorescence staining and RT-PCR. Radioimmunoassay was used to analyse anti-HBV activity of RAd/TRL.

RESULTSRecombinant RAd vectors were prepared successfully. Effective expression of RAd/TRL in HepG2.2.15 cells resulted in a significant decrease of HBsAg and HBeAg concentration in comparison with the controls.

CONCLUSIONAdenoviral vector-mediated targeted ribonuclease can effectively inhibit HBV replication.

Adenoviridae ; genetics ; Cell Line ; Cell Line, Tumor ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Genetic Vectors ; genetics ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonucleases ; genetics ; metabolism ; Transfection ; Virus Replication ; genetics

Cell Line ; DNA, Viral ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Humans ; Ribonucleases ; genetics ; Transfection ; Virus Replication

Objectives To explore the relationship between polymorphism in estrogen receptor alpha (ERα)gene Xba I and child dental fluorosis.Methods Qiulou township of Kaifeng and Sunying township of Tongxu counties of Henan province were chosen as the investigation spots in 2006.An area of water drinking endemic fluorosis(high fluoride area)and a non-endemic area(control area)were chosen in every spot,where dental fluorosis of children aged 8 to 12 years old were examined and diagnosed by using the Dean method.The children in the high fluoride areas were divided into dental fluorosis group and control group of the endemic areas according to dental fluorosis status,and the children in the control areas as control gruop of non-endemic areas.The Xba I polymorphism in the ERα gene was genotyped using the PCR-RFLP procedure.The fluoride levels in the urine samples from the three groups were detected by fluoride ion selective electrode and over standard rate of the urine was counted.Results The prevalence rate of dental fluorosis in high fluoride areas was 51.7%(74/143)and the community fluorosis index was 1.310.No dental fluorosis case was checked out in the control and the community fluorosis index was 0.021.The over standard rate of urine fluoride in dental fluorosis group[84.6%(121/143)]was significantly higher than that of control in non-endemic area[9.6%(9/94);χ2=125.95,P<0.01].The frequency distribution of ERα Xba I genotype was XX 6.8%(5/74),xx 36.5%(27/74),xx 56.8%(42/74)in dental fluorosis group;XX 15.9%(11/69),Xx 37.7%(26/69),xx 46.4%(32/69)in the eontrol of the endemic areas;XX 14.9%(14/94),Xx 43.6%(41/94),xx 41.5%(39/94)in children from the control in non-endemic area,respectively.No significant difference was found among the three groups(χ2= 3.450, P > 0.05). Allele frequency of ERα Xba I genotypes was X 22.7%(30/132), x 77.3%(102/132) in dental fluorosis group and X 35.5%(39/110),x 64.5% (71/110) in the control in endemic area when urine fluorosis of children was exceeding standard and significant difference was found in this two groups(χ2 = 4.768, P < 0.05; OR = 0.535,95% CI:0.305 - 0.941). Conclusion Children who carried X allele frequency of ERα Xba I genotypes have a lower risk of dental fluorosis when children with high-loaded fluoride status.

To study the impact of the enterovirus 71(EV71) on the nuclear transport mechanism,The pGFP-NLS vector with nuclear location signal(NLS) was constructed, RD cells transfected by the pGFP-NLS vector were inoculated with the EV71 or cotransfected by EV71-2A vector. The results showed that GFP protein with NLS was expressed in the cytoplasm due to the inhibition of nuclear transport. In order to further study the mechanism of the EV71 to prevent nuclear transport,Nup62 was detected by Western blotting after RD cells were infected with EV71 or transfected by EV71-2A vector. The results showed that decreased expression of Nup62 could be detected after infection with EV71 and transfection by EV71-2A vector. This study demonstrates that the cleavage of Nup62 by EV71 2A protease may be the mechanism of nuclear transport inhibition.
Active Transport, Cell Nucleus ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; metabolism ; Enterovirus Infections ; virology ; Gene Expression Regulation, Viral ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Membrane Glycoproteins ; metabolism ; Nuclear Localization Signals ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Peptide Hydrolases ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Transfection