Objective To investigate the expression of human bone morphogenetic protein-7(hBMP-7)in rat bone malTOW stromal cells(BMSC)with a recombinant adenovirai vector carrying the hBMP-7 gene(Ad-hBMP-7) and study the effecta of Ad-hBMP-7 transfection on BMSC difierentiation.in order to explore the possibility for hBMP-7 gene therpy.Methods The rat BMSC cultured in vitro.They were divided into 3 groups:untreated group,Ad-hBMP-7 and Ad-GFP transduced treated group.The rat BMSC were transfected by Ad-hBMP-7 and Ad-GFP. The expression of hBMP-7 was detected by RT-PCR and Western blot analysis.and the alkaline phosphatage (ALP)activity of the BMSC was observed.ResulIs In the Ad-hBMP-7 transduced treated group.hBMP-7 mRNA expression wag manifested detected by RT-PCR(470 bp),Westem blot analysis demonstrated that these cells indeed produced the hBMP-7 protein(Mr.15×103);10 days after transduction treatment,most of the BMSC were had brown black particles stained positively by ALP activity.But in Ad-GFP transduced treated group and untreated group they did not.Conclusions Ad-hBMP-7 could efficiently transfect BMSC and promote the conversion to osteoblast.The expression of hBMP-7 in rat BMSC provides a basis for hBMP-7 gene therapv.

OBJECTIVETo prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity.

METHODSRecombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay.

RESULTSELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%.

CONCLUSIONWe obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.

Animals ; Antibodies ; immunology ; Antibody Formation ; Egg Proteins ; immunology ; Female ; Immune Sera ; Male ; Membrane Glycoproteins ; immunology ; Mice ; Oocytes ; Rabbits ; Receptors, Cell Surface ; immunology ; Recombinant Proteins ; immunology ; Sperm-Ovum Interactions ; Spermatozoa ; Zona Pellucida Glycoproteins

OBJECTIVETo investigate the methylation frequencies of multiple tumor suppressor genes (TSGs) in hepatocellular carcinoma (HCC) and the clinical implication of aberrant DNA methylation in molecular carcinogenesis of HCC.

METHODSSixty samples of HCC and the paired adjacent liver tissue, 16 samples from post-hepatitis cirrhotic livers, 5 from livers with chronic hepatitis and 5 from normal livers were collected. Eight TSGs frequently silenced by hypermethylation of their promoters in various types of digestive tumors were selected, including APC, RASSF1A, p16, GSTP1, MGMT, DAPK, SOCS-1 and RIZ1. The status of promoter methylation in these 8 genes was investigated using methylation-specific polymerase chain reaction. The clinicopathological data of HCC were also analyzed in order to evaluate the clinical implication of aberrant methylation in HCC.

RESULTSMethylation of the 8 TSGs was quite frequent in HCC, with a methylation rate of 95.0% in RASSF1A, 90.0% in APC, 73.3% in GSTP1, 65.0% in p16, 61.6% in RIZ1 and 60.0% in MGMT. Methylation of the 6 genes was more frequent in HCC than that in adjacent tissues (P < 0.05). The methylation rate of MGMT, GSTP1 and RIZ1 in the adjacent tissues was 41.6%, 40.0% and 25.0%, respectively, significantly higher than that in cirrhotic liver (P < 0.05). p16 methylation was more frequently observed in HCC in elderly patients. The frequency of MGMT methylation was tended to be higher in giant HCC than that in the other types of HCC. Patients with MGMT methylation in the tumor were found to have a shorter disease free survival.

CONCLUSIONDifferent frequency of methylation in hepatocellular carcinomas, adjacent liver tissues and cirrhotic livers implies that epigenetic alteration in the hepatocellular carcinogenesis may be a gradually progressive process. Methylation status of MGMT, GSTP1 and RIZ1 may be promising in risk assessment of hepatocellular carcinoma and in early diagnosis. Furthermore, MGMT methylation might be also used as a potential prognostic biomarker for hepatocellular carcinoma patients.

Adult ; Aged ; Base Sequence ; Carcinoma, Hepatocellular ; genetics ; metabolism ; DNA Methylation ; DNA Modification Methylases ; genetics ; metabolism ; DNA Repair Enzymes ; genetics ; metabolism ; DNA, Neoplasm ; genetics ; DNA-Binding Proteins ; genetics ; metabolism ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Glutathione S-Transferase pi ; genetics ; metabolism ; Hepatitis B, Chronic ; genetics ; metabolism ; Histone-Lysine N-Methyltransferase ; Humans ; Liver ; metabolism ; Liver Cirrhosis ; genetics ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Molecular Sequence Data ; Neoplasm Recurrence, Local ; Nuclear Proteins ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism ; Young Adult

BACKGROUNDNerve growth factor (NGF) is well-known for its important role in the development and maintenance of the nervous system. Along with its neurotrophic role, NGF has been detected in the testis of mouse, rat and human, suggesting an additional non-neurotrophic effect in the male reproductive system. The expression of β-NGF in the undescended testes (cryptorchidism) has not been detected at present. The aim of this study was to evaluate the expression of β-nerve growth factor mRNA and protein in experimental cryptorchidism.

METHODSA unilateral mechanical cryptorchidism model in the Sprague-Dawley rat was established and the expression of β-NGF with histologic changes in experimental cryptorchidism were investigated using one step quantitative real-time reverse transcription-polymerase chain reaction, in situ hybridization histochemistry, immunofluorescence and hematoxylin-eosin staining.

RESULTSThe expression of β-NGF mRNA and protein were both significantly decreased in the development of unmarred testis and cryptorchidism-induced testis, and the decrease of β-NGF in cryptorchidism-induced testis was far greater than that in uninjured testis.

CONCLUSIONFrom this investigation, we confirmed a lower expression of β-NGF in undescended testes than in the development of testis.

Animals ; Cryptorchidism ; genetics ; metabolism ; Male ; Nerve Growth Factor ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influencing factors on tuberculosis (TB) in four provinces in the eastern and central parts of China.

METHODSFrom Nov. 2009 to Feb. 2011, three population-based field surveys were conducted among the resident population in several townships/streets in Guangdong, Hunan and Jiangsu provinces and Shanghai municipality to collect TB-related information. 474 sputum smear positive TB patients and 1896 controls were randomly selected from the population under study and each case was matched by province, age and sex using a frequency matching method. Single-variable and multiple non-conditional logistic regression modeling were applied for data analysis, and odds ratios (ORs) and their corresponding 95% confidence intervals (CIs) were estimated.

RESULTSData from Single-variable analysis showed that TB history, history of exposure to TB, DM history, immigrant population and per-capita living space were risk factors for TB, and high level of education was protective factors.

RESULTSfrom multiple logistic regression showed that the risk factors of TB would include the following items: history of having had TB (OR = 52.356, 95%CI: 18.956 - 144.607), living space over 50 m(2)per-capita (OR = 8.742, 95%CI: 1.107 - 69.064), history of exposure to TB (OR = 6.083, 95%CI: 2.336 - 15.839) and being immigrants (OR = 3.306, 95%CI: 1.907 - 5.734), while having had high degree of education as the protective factor of TB (OR = 0.284, 95%CI: 0.110 - 0.733).

CONCLUSIONControl programs targeting those ever having TB patients and contacts to TB patients as well as immigrants should be strengthened.

Adult ; Aged ; Case-Control Studies ; China ; epidemiology ; Cities ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Risk Factors ; Tuberculosis ; epidemiology

Objective To establish the surveillance and warning index system corresponnding with the epidemic characteristics of tuberculosis,in China.Methods Literature review and expert meeting were conducted to formulate the preliminary index system frame.Delphi method was used for screening the index and determining the weight of each index.Results Two-round Delphi consultations were performed.The activity coefficients were 87.0％,90.0％ respectively,with means of authority coefficient as 0.850 ± 0.055 and 0.917 ± 0.017,respectively.Mean scores of the index were 7.063 ± 1.435,8.156 ± 0.611 respectively,with the means of coefficient of variation as 0.352 ±0.161 and 0.170 ± 0.057 respectively.The harmony coefficients were 0.322 (x2=499.472,P＜0.05)and 0.393 (x2=241.126,P＜0.05) respectively.After the two-round consultation,the tuberculosis monitoring and warning index system was developed,including 4 first-class indicators,9 second-class indicators and 48 third- class indicators.Conclusion An index system was established for tuberculosis monitoring and early warning that could provide evidence for tuberculosis prevention and control as well as for the forecasting and warning model of the disease.