Objective To investigate the steroid levels of urine in Chinese athletes. Methods Subjects were 1217 Chinese athlete, 8 kinds of steroids named androsterone (An), etiocholanolone (Etio), dihydrotestosterone (DHT), dehydroepiandrosterone (DHET), Testosterone (T), Epitestosterone (ET), 5?-androstane-3?, 17?-diol (5?-diol), 5?-androstane-3?, 17?-diol (5?-diol) were measured by gas chromatography combined with mass spectrometry (GC/MS) after the urine sample being treated with chemical pre-extraction, hydrolysis, extraction and derivatization. Results The urine steroid levels of the Chinese male athlete were higher than those of the female athlete, but both male and female athlete had a lower urine steroids especially testosterone and 5?-androstane-3?,17?-diol, 4～7 times lower than other country's athletes. Some ratios of urine steroids such as An/Etio, T/ET, 5?-diol/5?-diol were relatively steady, which were affected less by gender.

Objective:To search for proteins interacting with ARA267-? with the yeast two-hybrid system in order to further investigate the function of ARA267-?. Methods:We screened a pretransformed human brain cDNA library with the pGBKT7-PHD-SET recombinant plasmid as a bait which express four PHD(plant homeodomain) and one SET[Su(var)3-9, Enhancer-of-zeste, Trithorax] conserved domains in ARA267-?.The plasmids in positive yeast clones were selectively identified by restriction analysis and DNA sequencing. The interactions were retested by yeast two-hybrid assay. Results: There were about six hundreds positive yeast clones on SD/-Ade/-His/-Leu/-Trp/2.5 mmol/L 3-AT/ X-?-Gal high-stringency selection plates. The pACT2-cDNA plasmids in sixty-five yeast clones were isolated and thirty-five cDNA inserts were sequenced. Sixteen different genes,including DR6(death receptor-6), PIAS3 (protein inhibitor of activated STAT3)and RanBPM(Ran-binding protein in the microtubule-organizing center), were identified after BLAST in GenBank. The yeast two-hybrid retest showed that all but RanBPM were true interactors of ARA267-?-PHD-SET. Conclusion: The ARA267-?-PHD-SET can interact with several distinct proteins. This suggests that ARA267-? is a protein having multiple functions. RanBPM might be a transcriptional factor.

Objective To make a survey of medication taken by Chinese athletes and a comparison between Chinese athletes and athletes from other countries in order to get information about how to improve Chinese athletes' performance. Method The information came from the forms"Doping Control Sample Collection" in which athletes answered the question: "What medications have you taken in the past 3 days?" The medicines taken by athletes were classified and statistically analyzed.Results 2,330 athletes and 25 kinds of sports were involved in. Medicines were statistically analyzed with 4 classes: profiling of declaration, vitamins and minerals, medicines for treatments, alternative medicine. Conclusion The survey recorded the types of supplements and medications taken by athletes in China in 1999. Chinese athletes took less vitamins and more alternative medicines than athletes from other countries.

Objective: To investigate the relationship between heart rate variability (HRV), blood pressure variability (BPV) and autonomic nerve function, blood vessel damage in patients with essential hypertension (EH) via synchronous monitoring. Methods: A total of 275 EH patients admitted to our hospital from 2011-04 to 2014-01 were enrolled. The vascular function was assessed by carotid-femoral pulse wave velocity (PWV). Based on PWV, the patients were divided into 2 groups: Normal PWV group (PWV<9m/s),n=185 and High PWV group (PWV≥9m/s),n=90. Synchronic 24h dynamic electrocardiogram (Holter) and 24h ambulatory blood pressure monitoring (ABPM) were performed in all patients. t-test, chi-square test, person liner correlation study and multi stepwise regression analysis were conducted to explore the relationship between HRV, PBV and PWV. Results: HRV and BPV in High PWV group had been changed unusually. Compared with Normal PWV group, High PWV group showed decreased standard deviation of the average of all normal-to-normal intervals in all 5-minute intervals (SDANN) (159.66±66.50) ms vs (194.36±119.29) ms and increased 24 h systolic blood pressure standard deviation (24h SSD) (14.40±3.65) mmHg vs (12.98±3.46) mmHg, all P<0.01; increased new index of night/day HR ratio (0.90±0.08) vs (0.87±0.06), P<0.01 and it had liner correlation to PWV (r=0.169, P=0.005). Multi stepwise liner regression analysis indicated that 24hSSD and HRV at low frequency (LF) portion had obvious and independent correlation to PWV (standard β value=0.352 and 0.212 respectively). Conclusion: ① EP patients were with decreased HRV (SDANN), increased BPV (24h SSD) and the higher incidence of arteriosclerosis; 24h SSD and HRV at LF portion were the most 2 important risk factors affecting PWV. ②Autonomic nerve dysfunction, vagus nerve over-excitatory were the independent risk factors for promoting the occurrence and development of arteriosclerosis in EH patients. ③Night/day HR ratio as a sensitive index for examining autonomic nerve function was independently related to hypertensive vessel damage. Synchronic monitoring of HRV and BPV is helpful to identify blood vessel damage in EH patients.

Objective To determine low-concentration clenbuterol in edible meat based on the solidphase extraction coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).Method The homogenated sample was acidized to remove proteins,and purified using the liquid-liquid extraction and MCX Oasis solid prepared extraction column,then further treated with gradient elution with the mobile phase of ammonium formate (10 mmol/L and pH 3.5) and acetonitrile.The clenbuterol was completely separated on Eclipse C18 (1.8 μm,4.6×100 mm) column and detected in multiple reaction monitoring (MRM) mode.Results A good linearity was achieved for clenbuterol in the arrange of 0.01～0.2 μg/kg based on the internal standard calibration of D9-clenbuterol,with the linearity correlation coefficient greater than 0.99 and the detection limit of 0.005 μg/kg.The relative recovery of target compounds spiked in blank sample at three levels ranging from 78.8 to 114.8％,with the relative standard deviations less than 10％.Conclusion The method in this research is simple,rapid,reliable and suitable to confirm low-concentration clenbuterol in edible meat.

Objective To evaluate immunogenicity of the recombinant protein GST-p55/570 of Pneumocystis carinii.Methods The fusion protein GST-p55/570 was expressed from the prokaryotic expression plasmid pGEX-570,and purified by using glutathione-agarose.The expressed product was analyzed by SDS-PAGE.Thirty-three mice were randomly divided into three groups,immunized with GST-p55/570,GST and PBS,respectively.Each group was immunized for four times at 2 week intervals.At the 7th day after final immunization,spleen was removed to obtain single cell suspension.Proliferation ability of lymphocytes was determined by MTT.Serum samples were collected at pre-immunizaton and two weeks after each immunization.Antibody level in sera of mice was determined by ELISA.The immune response to the recombinant GST-p55/570 recognized by sera of immunized mice was examined by Western blotting.Results The expressed fusion protein GST-p55/570 showed a Mr 47 000.Compared with GST group(1.134 5?0.073 5) or PBS group(1.124 8?0.041 6),a higher stimulation index(2.063 0?0.160 2) was revealed in GST-p55/570-immunized mice(P

Objective To explore the association between SLC2A9,SLC17A3,ABCG2 single nucleotide polymorphisms and gout susceptibility in Quanzhou.Methods One hundred and fifty-four cases of gout patients and 160 healthy controls were selected,single nucleotide polymorphisms (SNP) of SLC2A9 SLC17A3,ABCG2 with tri-primer polymerase chain reaction (PCR) were tested and the relation between different genotypes and primary gout prevalence were analyzed.Results High risk genotype frequency of rs16890979 was 93.5％ and 70.0％ in patients and healthy people,respectively (the difference of genotype frequency between the two groups was statistically significant (x2=55.377,P＜0.01).High risk allele frequency was 79.9％ and 48.4％ in patients and healthy people,respectively (allele frequency in different population was statistically significant,x2=67.128,P＜0.01).High risk genotype frequency of rs2231142 was 68.8％ and 38.7％ in patients and healthy people,respectively (the difference of the genotype frequency was statistically significant,x2=29.129,P＜0.01);High risk allele frequency was 43.5％ and 23.4％ in patients and healthy people,respectively (the difference of allele frequency was statistically significant,x2=28.468,P＜0.01) ; rs1165205was a protective SNP,low risk genotype frequency was 42.2％ and 45.6％ in patients and healthy people,respectively (the difference of genotype frequency was statistically significant,x2=0.373,P=0.571); High risk allele frequency was 26.0％ and 28.1％ in patients and healthy people,respectively (the difference of allele frequency was not statistically significant,x2=0.270,P=0.364).Conclusion SNP loci rs16890979 of SLC2A9 gene and rs2231142 of ABCG2 gene can be used as genetic markers for primary gout susceptibility in the Quanzhou area,but SNP loci rs1165205 of SLC17A3 gene has little correlation with the prevalence of primary gout in Quanzhou residents.

Objective To construct prokaryotic recombinant expression plasmid carrying Pneumocystis carinii Mr 55 000 antigen(p55) gene fragment and express the recombinant protein. Methods P. carinii pneumonia(PcP) rat models were established by subcutaneous injection of dexamethasone for 14 weeks. Total RNA was extracted from lung of P. carinii rat and p55 antigen gene fragment was cloned by RT-PCR,which was identified by sequencing. The 690 bp fragment was cloned to pGEX-4T-1,the recombinant plasmid was screened and identified by restriction analysis and PCR. The recombinant plasmid was finally induced with IPTG to express a new fusion protein,and the products were analyzed by SDS-PAGE and Western blot. Results A fragment of 690 bp was obtained by RT-PCR. The recombinant pGEX-4T-1/690 was constructed. SDS-PAGE revealed that the molecular weight of the recombinant protein was approximately Mr 62 000,the maximum amount of the fusion protein produced was 11.6% of the total protein. The recombinant protein can be recognized by GST antibody and by the sera from P. carinii infected rats using Western blotting. Conclusion Prokaryotic expression plasmid pGEX-4T-1/690 has been constructed and the recombinant fusion protein shows antigenicity.

Objective To evaluate the safety and efficacy of intra-arterial infusion neoadiuvant chemotherapy in local advanced bladder cancer. Methods Nineteen cases with T2-T4a bladder cancer were enrolled in this study.Intra-arterial infusion chemotherapy with Gemcitabine and Cisplatin (GC)were performed for 1 to 3 cycles before radical cystectomy.Postoperative values of hematological parameters,maximum diameter of tumors,TNM(tumor,node and metastasis)stages and pathological grades were compared with preoperative parameters of the same case. Results Compared to the values before GC chemotherapy,WBC count showed no significant change post-operative,(6.63±2.58)×109/L vs(5.12±2.91)×109/L(P=0.13);RBC(4.41+0.52)×1012/L vs(3.92±0.42)×1012/L(P=0.00)and platelet count(220.50±59.86)×109/L vs(157.05±56.72)×109/L(P=0.001)showed significant decrease;ALT did not show significant decrease(20.00±8.31 vs 26.88±17.04 U/L,P=0.08);Creatltme also showed no significant change(95.82±14.57 vs 88.04±17.76μmol/L,P=0.06);Maximum diameter of tumors decreased significantly(3.72±1.23 vs 2.80±1.29 cm,P=0.02).Compared with clinical TNM stages,pathological TNM stages demonstrated significant decrease in 9 cases;While cell differentiation did not show decrease. Conclusions Intra-arterial infusion with GC regimen can reduce tumor size,decrease TNM stages,while not causing significant adverse impact to radical cystectomy.Bladder-spare treatment is an option for chemotherapy-sensitive cases.

Objective To observe the effects of GABA on proliferation, cell cycle and expression of MMP-2, MMP-9 of pancreatic cancer cell line SW1990. Methods The effects of different concentration of GABA (0 ～ 320 μmol/L) on proliferation and cell cycle of pancreatic cancer cell line SW1990 was investigated by MTT assay and flow cytometry analysis, respectively. Expressions of MMP-2 and MMP-9 proteins were evaluated by Western blot analysis. Results GABA could promote the proliferation of SW1990 cells and influence the distribution of cell cycle, which made less cells of G0/G1 phase and more cells of S and G2/M phase. The value of A570 after GABA pretreatment at a dose of 320 μmol/L was 1. 11 ± 0.03, which was significantly higher than that in the control group (0. 56 ± 0.01, P < 0. 01 ), the cells of G0/G1 phase was (46.18 ± 1.12 )% ,which was significantly lower than (87.29 ± 1.34)% in the control group (P < 0. 01 ) ;the expressions of MMP-2 mRNA, MMP-9 mRNA and their proteins were 8.6, 6.8, 10.5, 8.4, respectively, which were significantly higher than those in the groups of the doses of 0 ～ 40 μmol/L ( P < 0. 05 ). Conclusions GABA could influence the proliferation and expression of MMP of SW1990 cells.