AIM　To establish an allele specific PCR amplification (ASA-PCR) for determination of the genotype of CYP2D6*10B polymorphism in Chinese subjects. METHODS　CYP2D6*10B alleles of 65 healthy Chinese subjects were analyzed by a two-step PCR assay and the correlation of genotype and phenotype was studied. RESULTS　There were 20 CYP2D6*10B heterozygous genotypes subjects (wt/m) in 35 very extensive metabolizers (VEMs), which consisted the major part of VEM subjects (57%). Meanwhile, 20 subjects consisting 69% of 29 intermediate metabolizers were CYP2D6*10B homozygous mutant genotypes (m/m). The poor metabolizer was also m/m. The metabolic ratio of CYP2D6*10B m/m subjects were larger than wt/m and wild type, the values were -1.49±0.54, -2.20±0.49 and -2.47±0.61 (P<0.01). CONCLUSION　PCR-ASA was shown to be a rapid and specific method. It can be used to study the genetic polymorphism, especially CYP2D6 intermediate metabolism.

Objective To construct a replication-deficient recombinant adenovirus expression vector of human heparanase (hpa). Methods The hpa gene was cloned at the downstream of CMV promoter of the adenoviral shuttle plasmid pDC315 in sense direction, and the resultant recombinant plasmid pDC315-hpa was transfected into HEK293 cell together with plasmid pBHGlox (deltaE1,3) containing adenoviral genome, then the replication-deficient recombinant adenovirus expression vector of hpa (Ad-hpa) was obtained, and it was identified by infection test, electronic microscope observation and PCR amplification. Results After purification and concintration,the titer of Ad-hpa reached 5?10 10pfu/ml. Virus particles could be found in virus concintration solution, and replication of virus was observed in HEK293 cells was observed under transmission electron microscope. Both adenovirus and hpa special fragment could be amplified from Ad-hpa by PCR, whereas hpa special fragment could not be amplified from the control. Conclusion The replication-deficient recombinant adenovirus expression vector of hpa was constructed successfully. This study established a foundation for further study on hpa vaccines and gene therapy for carcinoma.

Objective To investigate the changes in biologic behavior of dendritic cells (DCs) after being modified by human telomerase reverse transcriptase (hTERT) gene. Methods In order to obtain the hTERT gene modified DCs, DCs were transfected with a replication-deficient recombinant adenovirus expression vector of hTERT. Then the expression to hTERT and PCNA was assessed by by Western blot, mature markers on DCs surface were detected by flow cytometry, and the proliferative capacity was determined by MTT method. Results Immunohistochemical staining and Western blotting showed that the expression of hTERT was upregulated obviously in DCs after being modified by hTERT gene. Flow cytometry indicated that the expression of CD83 and CD86 remained unchanged. The DC growth curve showed that the number of DC-hTERT was increased slightly in the first 2 weeks, meanwhile the number of DC/rAd-LacZ and DC was decreased obviously. Although the number in 3 groups was all decreased in the third week, the number of DC/rAd-hTERT was still greater than the other control groups. It was also found that the expression of PCNA was increased in DC/rAd-hTERT compared with that of immature DC, mature DC or DC/rAd-LacZ by Western blot. Conclusion After rAd-hTERT modification, life span of DC in vitro is extended and proliferative capacity is enhanced.

Objective The aim of this study was to explore the effect of androgen deficiency on serum hormone levels, visceral fat accumulation and inflammatory gene expression in miniature pigs fed a high-fat diet ( HFD) . Methods Sexually mature male Chinese Wuzhishan miniature pigs were divided into three groups ( animals/group) as follows:intact male pigs ( SHAM) , castrated male pigs ( CAS) and castrated male pigs plus testosterone treatment ( CAS+T) . The pigs were fed a HFD diet for 12 weeks. Serum levels of testosterone and leptin were measured and visceral fat were dissected and weighted. qRT?PCR was performed to determine the mRNA expression levels of lipogenic, lipolysis and inflammation relat?ed genes. Results (1) Serum testosterone levels were significantly decreased but serum leptin levels were significantly in?creased in the castrated pigs. These effects were recovered after testosterone treatment. ( 2 ) Visceral fat percentage was significantly increased in the castrated pigs, and testosterone treatment reduced the increased visceral fat in the castrated pigs. (3) Castration and testosterone treatment had no significant effects on the expression levels of lipogenic genes (FAS and ACC) and lipolysis genes (HSL and ATGL) in pigs fed a HFD. (5) Castration significantly induced the expressions of inflammatory genes including Leptin, CD68, CCL16, CCL23 and SAA, and testosterone treatment recovered the expres?sions of the above genes in the castrated pigs. Conclusions Castration?induced testosterone deficiency promotes visceral fat accumulation and upregulates the expression levels of inflammatory genes in miniature pigs fed a HFD. Moreover, tes?tosterone treatment ameliorates castration?induced visceral fat accumulation and inflammatory response in HFD?fed pigs.

Objective To investigate current health status of medical professionals in Guangdong Province to provide evidence for individualized health management. Methods Based on multi-stage stratified sampling method,3872 medical professional from 56 hospitals in Guangdong Province were enrolled and completed a self-designed questionnaire. The results were collected and compared using descriptive analysis, K-S test, Chi-square test and multivariate Logistic regression. Results A total of 3687 (95.2%) feed-backs were collected,of which 3502 (95.0%) could be used. Insomnia was the most serious problem among nurses (68. 9% ). The incidence rate of sub-health condition was 58. 2%. First-line clinical work,female, long working experience, tertiary hospital, no physical activity and insomnia were correlated with more serious sub-clinical health. Conclusion Unhealthy life style and sub-heahh problems negatively impact health of medical professional. Therefore, supervisors should pay more attention to those with certain characteristics.

A comparison of the number of clinicians per 1000 residents in the province,and analysis of their distribution and characteristics identified an imbalance of such resources between regionsand between urban and rural areas.A further analysis of the payment flow of medical insurance fund of the counties and districts probed into the deployment efficiency of clinical in the province,holding that the hck of a human resources deployment and guidance mechanism for clinical resources is key to toor healthcare equity and difficulties in accessibility of healthcare service.

Air ; analysis ; Air Pollutants ; analysis ; Hazardous Substances ; analysis ; Spectrophotometry, Atomic ; methods

Objective To investigate alteration of SHP-1 mRNA and protein of bone marrow cells of leukemia mice after ?-ray exposure.Methods A total of 318 BALB/c mice were exposed to ~(60)Co ?-ray once a week for 4 weeks,and the total dose of ?ray received by mice was 7.00 Gy.By pathological examination,39 mice developed thymic lymphoma,14 acute lymphocytic leukemia,21 T-lymphoblast leukemia/lymphoma,1 spiroma,4 malignant yolk sac tumor.Exposed to ?-ray,the mice that developed leukemia or were free of canceration were used in our study(n=10 in each group),and another 10 mice free from irradiation served as control.SHP-1 mRNA and protein in femoral bone marrow cells were detected by real-time fluorescence quantitative PCR(FQ-PCR) and Western blotting respectively.Results SHP-1 mRNA in leukemia mice was(5.26?2.14),significantly higher than that of mice free of canceration(3.68?1.27) and controls(2.95?1.09)(P

Objective To investigate alteration of SHP-2 mRNA and protein of bone marrow cells of leukemia mice after ?-ray exposure.Methods A total of 318 BALB/c mice were exposed to ~(60)Co ?-ray once a week for 4 weeks,and the total dose of ?-ray received by mice was 7.00 Gy.By pathological examination,39 mice developed thymic lymphoma,14 acute lymphoblastic leukemia,21 T-lymphoblast leukemia/lymphoma,1 spiroma,4 malignant yolk sac tumors.Exposed to ?-ray,the mice that developed leukemia or were free of canceration were used in our study(n=10 in each group),and another 10 mice free from irradiation served as control.SHP-2 mRNA and protein in femoral bone marrow cells were detected by real time fluorescence quantitative PCR(FQ-PCR) and Western blotting respectively.Results SHP-2 mRNA was(5.08?2.87) in leukemia mice,(4.59?2.36) in mice free of canceration and(3.54?1.02) in controls.SHP-2 protein was(0.956?0.125) in leukemia mice,(0.892?0.236) in mice free of canceration and(0.712?0.368) in controls.The enzyme catalytic activity of SHP-2 was(0.156?0.069),(0.118?0.065),(0.098?0.048).No significant difference in mRNA,protein or enzyme catalytic activity of SHP-2 was found in leukemia mice,mice free from canceration and control mice.Conclusion SHP-2 mRNA and protein was significantly increased in bone marrow cells of leukemia mice induced by ?-ray irradiation.

Objective To investigate the alterations of SHP-2 mRNA of thymus cells of mice with ? ray-induced leukemia. Methods BALB/c mice were randomly divided into canceration group, non-canceration group, and control group. The gene mutation and content of SHP-2 mRNA in thymus cells were detected by PCR-SSCP, DNA sequencing, and real time fluorescence quantitative PCR (FQ-PCR). Results PCR-SSCP showed there was gene mutation within 700～1 096 bp of SHP-2 mRNA in thymus cells of mice in the canceration group, which was not supported by DNA sequencing. No significant difference in change of SHP-2 mRNA content was found in thymus cells in canceration, non-canceration, and control groups. Conclusion There is translational abnormality of SHP-2 mRNA in thymus cells of mice with ? ray-induced leukemia, which is presented as translational enhancement.