OBJECTIVEEstablishment of determination method of carbon disulfide in charcoal tube with low toxicity solvents desorption-gas chromatography.

METHODSFour types of solvent with low toxicity are applied respectively as substitution of benzene to desorb the carbon disulfide in samples of charcoal tube. The signal strength and desorption efficiency of the detector are compared by using different solvents.

RESULTSChloroform has been considered as the best alternative solvent of benzene. Carbon disulfide has a good linearity (R = 0.9997) over the concentration of 0 ∼ 54.7 µg/ml, detection limit can reach 0.2 µg/ml. When the sampling volume is 3.0 L, the minimum detectable concentration is 0.07 mg/m(3).

CONCLUSIONWith the use of chloroform, the health hazard to laboratory personnel and environment pollution as well as the costs of experiments are reduced.

Air Pollutants, Occupational ; analysis ; Carbon Disulfide ; analysis ; Chromatography, Gas ; methods ; Solvents ; analysis ; Workplace

AlM: To silent hypoxia inducible factor-1α ( HlF-1α) gene in malignant melanoma of the choroid cell by small interference RNA ( siRNA ) and investigate its effect on the expression of matrix metalloproteinase-2 ( MMP-2 ) in the choroid cell line human uveal melanoma cell (OCM-1) in hypoxia environment.METHODS:OCM-1 cells cultured on culture flask were divided into normal group and hypoxia group. Hypoxia group were divided into five groups: simple hypoxic group, and interference group, and negative control group, and positive control group, and liposome group. Normal group cells were cultured on DMEM culture flask with 10% FBS, 100U/mL penicillin, 100μg/mL streptomycin as well as high concentration of glucose. The cells were maintained at 37℃ in a humidified 5% CO2 incubator. Cells in good condition were selected for experiment. For hypoxia group, chemical hypoxia inducer CoCl2 was added into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment. We designed and synthesised siRNA ( siRNA + negative control+positive control ) , the target sequences of the HlF-1α to transfect hypoxic malignant melanoma of the choroid cell. SiRNA including HlF-1α siRNA, β-actin siRNA and negative control group synthesized in vitro transfected hypoxic OCM - 1 cell through Lipofectamine2000. The expression of HlF-1α, MMP-2 gene and the protein were detected by RT-PCR and Western blot. RESULTS: Compared with the normal group, the expression of HlF-1α mRNA was not obviously changed (P>0. 05), but the expression of HlF-1α protein and MMP- 2 mRNA protein was significantly higher ( P<0. 05) . Compared with the other hypoxia groups,β-actin mRNA expression of positive control group decreased ( P< 0. 05 ) , which proved successful transfection. The expression of HlF-1α mRNA and the expression of its protein and both MMP-2 mRNA and its protein was significantly lower ( P < 0. 05 ). The negative control group, liposome control group had no significant difference in the detection of factors (P>0. 05).CONCLUSlON: Hypoxia status may upregulate the HlF-1α in OCM-1 cells by increasing the expression of protein. Hypoxia can also inactivate MMP-2, resulting in upregulation of MMP-2 RNA and the expression of its protein. The expression of HlF-1α and MMP-2 mRNA can be down-upregulated by transfecting OCM-1 with HlF-1α siRNA.

OBJECTIVETo study the biological exposure index of carbon disulfide in China.

METHODSHigh-performance liquid chromatography (HPLC) was used to detect the levels of 2-thiothiazolidine-4-carboxylic acid (TTCA) in the urine of the workers after working shift end, Gas chromatography was used to detect the concentrations of the carbon disulfide in the workplace air. The relationship between the urine TTCA levels and the concentrations of the carbon disulfide was analyzed, the biological exposure index and judgement result from PC-TWA were compared.

RESULTSThe levels of TTCA in urine of workers occupationally exposed to carbon disulfide were closely and positively related with the concentrations of the carbon disulfide in the workplace air. The regression equation was Y = 0.265X - 0.165, The biological exposure index of carbon disulfide were calculated by regression equation according to occupational exposure limits of carbon disulfide in China.

CONCLUSIONThe biological exposure index of CS(2) in China might be revised for 1.2 mg/g Cr.

Carbon Disulfide ; analysis ; Chromatography, Gas ; Environmental Monitoring ; Humans ; Occupational Exposure ; analysis ; Thiazolidines ; urine ; Threshold Limit Values ; Workplace

OBJECTIVEEstablishment of determination method of 2-thiothiazolidine-4-carboxylic acid (TTCA) in urine with HPLC.

METHODSA volume of 0.5 ml hydrochloric acid (2 mol/L) and 0.5 ml pure water was added into 1 ml urine, and then extracted by 4 ml of diethyl ether by shaking for 2 min. Remove the water phase in a tube with plug and extract again, mix the two extraction diethyl ether together, take 4 ml by adding 2 ml borax-monopotassium phosphate buffer and shaking for 2 min to extract, then take the water phase to detect. A C(18) column and UV detector were used for separating and detecting. The wavelength was 273 nm, the flow rate was 1.0 ml/min, and the injection volume was 20 µl.

RESULTSTTCA has a good linearity (r = 0.9995) over the concentration of1 1 ∼ 10 µg and the minimum detectable concentration of TTCA in urine was 0.1 µg/ml. The within-day precision (RSD) were 8.4%, 3.0% and 1.7%, the between-day precision (RSD) were 11%, 3.8%, 1.9%, respectively. The extraction recovery were between 80% ∼ 102%.

CONCLUSIONThe method was accurate and sensitive to detect TTCA in urine.

Carbon Disulfide ; urine ; Chromatography, High Pressure Liquid ; methods ; Humans ; Thiazolidines ; urine

OBJECTIVETo investigate the molecular genetics basis for a novel HLA allele, HLA-B*5614, in Chinese population.

METHODSDNA was extracted from whole blood by salting-out method. The HLA-B exons 2-4 of the proband was amplified and the amplified product was cloned using TOPO cloning sequencing kit to split the two alleles apart. Both strands of exons 2,3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing.

RESULTSThe sequencing results showed the HLA-B alleles of the proband as B*1502 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY601726, AY610727, AY610728). After BLAST analysis, the novel allele differs from B*5608 by a single nucleotide at position 277G-->C in exon 2. This results in an amino acid change from Gly to Arg at codon 93.

CONCLUSIONThis allele is a novel allele and has been officially named B*5614 by the WHO Nomenclature Committee.

Alleles ; Exons ; genetics ; HLA-B Antigens ; genetics ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA ; methods

OBJECTIVETo study the efficacy and safety of Eviprostat for the treatment of benign prostatic hyperplasia (BPH).

METHODSAn open, multicentral clinical trial was conducted in 100 patients with BPH. Patients received a 12-week oral administration of Eviprostat 2 tablets per-time, 3 times a day. The main indexes of efficacy include international prostatic symptom score (IPSS), maximum urinary flow rate (Qmax), residual urine ( Ru) and prostatic volume (V). The additional indexes are quality of life score (QOL) and average urinary flow rate (Qave).

RESULTSAfter a 12-week therapy, IPSS, QOL score, Qmax and Qave were significantly improved. IPSS was averagely decreased by 5.67 (P < 0.001); QOL score was averagely decreased by 1.44 (P < 0.001); Qmax was averagely increased by 1.70 ml/s (P <0.001); Qave was averagely increased by 1.15 ml/s (P < 0.001); Ru was averagely decreased by 5.07 ml (P = 0.046) , PSA level was averagely decreased by 0.129 microg/L (P < 0.017). The clinical adverse event rate was 1%.

CONCLUSIONEviprostat is a kind of safe, effective and preferable drug for treating BPH. It can improve the subjective symptoms and objective measures of the patients.

Aged ; Aged, 80 and over ; Drug Combinations ; Ethamsylate ; adverse effects ; therapeutic use ; Humans ; Male ; Middle Aged ; Plant Extracts ; adverse effects ; therapeutic use ; Prostatic Hyperplasia ; drug therapy ; Quality of Life ; Treatment Outcome ; Urodynamics

OBJECTIVETo investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.

METHODSGenomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.

RESULTSThe sequencing results showed HLA-DRB1 alleles of the proband as DRB1*090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1*120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.

CONCLUSIONThis allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis

OBJECTIVETo investigate the effect of Salvia miltiorrhiza on glomerulosclerosis induced by Ang II.

METHODRat mesangial cells were exposed to 100 nmol L(-1) Ang II. Meanwhile, we added S. miltiorrhiza injection of different concentrations to Mcs. PAI-1 mRNA and protein, TGF-beta1 in serum free MEM medium, and the level of cellular reactive oxygen species (ROS) were measured.

RESULTS. miltiorrhiza notably attenuated Ang II induced expression of PAI-1 in a concentration-dependent manner. Meanwhile, S. miltiorrhiza suppressed the production of TGF-beta1 and cellular ROS in mesangial cells.

CONCLUSIONS. miltiorrhiza can alleviate glomerular sclerosis. The renoprotective effects of S. miltiorrhiza may be due to its ability to decrease Ang II -induced PAI-1 and TGF-beta1 secretion and cellular ROS level.

Angiotensin II ; pharmacology ; Animals ; Blotting, Western ; Cells, Cultured ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Gene Expression ; drug effects ; Injections ; Mesangial Cells ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; Plasminogen Activator Inhibitor 1 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; chemistry ; Transforming Growth Factor beta1 ; blood ; secretion

The prognostic value of phosphatidylinositol-4,5-bisphosphate 3-kinase,catalytic subunit alpha (PIK3CA) in patients with esophageal squamous cell carcinoma (ESCC) is controversial.We aimed to investigate the prognostic significance of PIK3CA mutation in patients with ESCC.EMBASE,PubMed,and Web of Science databases were systematically searched from inception through Oct.3,2016.The hazard ratios (HRs) and 95％ confidence intervals (CI) were calculated using a random effects model for overall survival (OS) and disease-free survival (DFS).Seven studies enrolling 1505 patients were eligible for inclusion of the current meta-analysis.Results revealed that PIK3CA mutation was not significantly associated with OS (HR:0.90,95％ CI:0.63-1.30,P=0.591),with a significant heterogeneity (I2=65.7％,P=0.012).Additionally,subgroup analyses were further conducted according to various variables,such as types of specimen,the sample size,technique and statistical methodology.All results suggested that no significant relationship was found between PIK3CA mutation and OS in patients with ESCC.For DFS,there was no significant association between PIK3CA mutation and DFS in patients with ESCC (HR:1.00,95％ CI=0.47-2.11,P=0.993,I2=73.7％).Publication bias was not present and the results of sensitivity analysis were very stable in the current meta-analysis.Our findings suggest that PIK3CA mutation has no significant effects on OS and DFS in ESCC patients.More well-designed prospective studies with better methodology for PIK3CA assessment are required to clarify the prognostic significance of PIK3CA mutation in ESCC patients.

Objective To understand the status on AIDS awareness,AIDS-related behaviors,risk factors on HIV infection status among 15-90 years or older men at the sexually transmitted disease clinics.Methods Data from the 2009 and 2010 national sentinel surveillance system,regarding men attending sexually transmitted disease clinics was collected from Guangdong,Guangxi,Henan,Sichuan,Yunnan and Jiangxi provinces,where the AIDS epidemic among 15-90 years or older population was serious.Data was uploaded to National Center for AIDS/STD Control and Prevention (NCAIDS) through the comprehensive AIDS control and prevention information system.Data was then analyzed by SPSS 18.0.Results A total of 64 003 pieces of data were collected.Among them,8783 ( 13.7％ ) were related to men at ≥ 50 years or older,and to men 15-49 years older were 55 220.The rates on the awareness of AIDS knowledge were from 69.6％ vs.80.1％,on frequently having had commercial sexual contacts in the last three months were between 34.1％vs.36.6％,on having had casual sexual contact in the last three months were 18.7％ vs.28.4％,on having had homosexual anal intercourse as 0.7％ vs.1.4％.The rates of taking HIV antibody testing in the last year (14.3％ vs.17.1％ ) among this population were all significantly lower than the rate among the 15 to 49 years age group.However,the HlV-positive rate among the older age group (fifty years of age or older) was significantly higher than the rate among 15 to 49 year age group ( 1.1％ vs.0.7％ ).Regard the fifty years of age or older men.Data from the multivariate logistic regression analysis demonstrated that factors as:having regular partner (OR=0.588,P=0.034),having homosexual anal intercourse (OR=5.226,P=0.006) were associated with positivities of HIV antibody.Conclusion High-risk sexual behaviors,including homosexual anal intercourse were the major risk factors for men at ≥50 years or older age,related to the infection of HIV.