Acute Disease ; Adult ; Humans ; Hydrogen Sulfide ; poisoning ; Male ; Poisoning ; pathology

Objective To study the genomic molecular organization and genogroup of human metapneumovirus(hMPV) infected infants in Guangzhou of China. Methods Primers were designed on the basis of the genomic sequence of hMPV 00-1 strain(AF371337) in the GenBank, and amplify hMPV genomeby RT-PCR. The PCR-products were cloned to T vector and sequenced, the genomic nucleotide sequences were analyzed with the programs Clustal W/X, DNASTAR and MEGA4. 1. Results The cloned strainhMPVgz01 genome is 13 327 bp in length, the genome contains eight open reading frames in the order 3-N-P-M-F-M2-SH-G-L-5. The genomic sequences of hMPVgz01 strain are compared with those of hMPV in GenBank, revealed that the homology with hMPV group A ranges between 92%-97%, homology with group B is 81%, and with avian metapneumovirus group C is 71%, the highest homology is with BJ1887 strain of genogroup A2b. The N, F, G genes of hMPVgz01 strain are compared with those corresponding genes of hMPV subgroups A1, A2, B1, B2, revealed that the highest homology is also with genogroup A2b. Conclusion The complete nucleotide sequence of hMPVgz01 strain isolated from Guangzhou in China is 13 327 bp in length, GenBank accession No. is GQ153651. Comparison of the genomic sequence and three genes of hMPVgz01 strain with those corresponding sequences of hMPV show the highest homology is with genogroup A2b. Sequence and phylogenetic analysis of the hMPVgz01 strain revegled that this isolate belongs to genogroupA2b.

Objective:To study the mutation frequencies of the exon 5 and the exon 8 of PTEN (phosphatase and tensin homology deleted on chromosome ten) gene in gastric carcinoma and investigate the relationship of the gene mutation and pathological differentiation and clinical stage.Methods:The mutation of exon 5 and exon 8 of PTEN gene was detected in 42 gastric carcinoma samples and the matched adjacent normal gastric mucosa with polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) method.The PCR products of mutant samples were analysed by DNA sequencing technique.Results:The mutation of PTEN was shown in 3 of the 42 gastric carcinoma tissues and in none of the adjacent normal tissues.The mutation rates of PTEN gene in poorly differentiated and well differentiated samples were 12.00% and 0,respectively (P0.05).The mutation rates of PTEN gene in clinical stage Ⅰand Ⅱ (5.88%) had no significant difference with that in clinical stage Ⅲand Ⅳ (8.00%) (P0.05).Conclusion:PTEN gene mutation occurs mainly in poorly differentiated gastric carcinoma tissues,and the mutation rate is not related to pathological differentiation and clinical stage.

Objective For realizing the simultaneous detection of multiple pathogens,by looking for a method with a combination of the new gene chip detection system based on nano-gold with the technology of restriction endonuclease without PCR.Methods Helicobacter pylori,Mycoplasma pneumoniae,Chlamydia trachomatis,Candida albicans,Ureaplasma urealyticum,and EB virus were selected as the experimental targets.Endonuclease Hha Ⅰ was selected as tool enzyme.After bering digested by Hha Ⅰ,the digested fragments of samples were tailed with poly-A.The samples were then detected by the gene chip detection system based on nano-gold.Both specific and common probes were used in the hybridization.The coincidence rate of the detection results between the new constructed chip test and the fluorescence quantitative PCR test in 168 clinical samples was examined.The stability and sensitivity of chips detection were also checked.Results The new constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR could be used to detect the target pathogens.The coincidence rate of the chip detection test and fluorescence quantitative PCR test in 168 clinical samples was 89.2%.Chip detection results showed that the stability of chips detection was 100% and the sensitivity was 50pmol/L.Conclusion The newly constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR can be widely applied in the simultaneous detection of Mycoplasma,Chlamydia,fungus,virus and bacteria.It shows a bright prospect in increasing the throughput of identifieation of pathogene.

0.05).EBV-specific IgM was detected more frequently in patients than that in controls(P

ObjectiveTo investigate glutathione S-transferase GSTO 1 Glu155△Glu genetic polymorphism and risks of arsenic poisoning caused by coal-burning in Guizhou.Methods GSTO1 Glu155 △Glu gene polymorphism was analyzed by polymerase chain reaction-with confronting two-pair primers among one hundred and thirty arsenic poisoning patients and one hundred and thirty healthy controls.The results were verified by DNA sequencing.The association between different genotypes and arsenic poisoning was analyzed by unconditional Logistic regression model.ResultsThe results of Glu/Glu and Glu/△Glu genotype detected by this method were consistent with those of DNA sequencing.The frequencies of GSTO1 Glu/Glu genotype and Glu/△Glu genotype were 94.85％(92/97) and 5.15％(5/97) in the patients,99.15％(117/118) and 0.85％(1/118) in the controls,respectively.The difference was statistically significant(x2 =3.896,P ＜ 0.05).△Glu/△Glu genotype was not found in both patients and controls.After age and sex adjusting,GSTO1 Glu155 △Glu polymorphism was found to be a risk factor of arsenic poisoning [odds ratio (OR) =1.85,95％ confidence interval (CI):1.39 - 17.48].ConclusionsThe study finds that GSTO1 Glu 155 △ Glu polymorphism is associated with risk of arsenic poisoning.The relationship between them should be further studied through increasing sample size.

The vip3A gene of Bt9816C was cloned and the sequencing result was submitted to GenBank (accession no.AY945939). The gene was identified as a novel vip3Aa gene, which was assigned name vip3Aa18 by the Bacillus thuringiensis delta-endotoxin nomenclature committee. Subsequently, vip3Aa18 was expressed in Escherichia coli BL21 and bioassay demonstrated that the purified recombinant Vip3Aa18 had high toxicity against Helicoverpa armigera and Spodoptera exigua. The results of sequence analysis revealed that a carbohydrate binding domain exists on the C-termini 536 to 667 residues of Vip3Aa18,which maybe participate in binding to midgut receptors in susceptible insects. Moreover, a transmembrane helices located on N-termini 272 to 292 residues was proposed responding for pore formation. Furthermore, a putative disulfide bond was found in the Vip3Aa18 sequence. The specific structures and sites of Vip3Aa18 sequence imply potential function.

Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P

Macrolactins are 24-membered macrolides produced by unidentified marine bacterium, Actinomadura sp. and Bacillus sp., which exhibit both antibacterial and antitumor activities in vitro. The environmental strain X-2 which was isolated from the sediment of the East China Sea produce Macrolatin A, B and O. In this study, a set of degenerate oligonucleotide primers, designed for amplification ketosynthase(KS) domains, had been employed to identify KS gene fragments of the X-2 DNA samples. One 645 bp KS fragment(GenBank accession no. EF486351)had been cloned and used as a probe to screen the genome DNA fosmid library of X-2. Three positive clones were selected and sequenced, Homologous analysis and the function prediction of the obtained PKS gene fragments suggested that macrolactin is the Polyketide Biosynthesis Product of the gene cluster obtained in the environmental strain X-2.

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.