4 weeks of arrested embryos.The AAH expression was found to have the similar result as HIF-1?'s.Conclusions The expression level of HIF-1? and AAH in villi of missed abortion patients is much lower than that of normal early pregnant women.HIF-1? and AAH have a function of supporting normal pregnancy,so their low expression may be an important cause of missed abortion.

Identification and validation of a new target is one of the most important steps for new antituberculosis (TB) drug discovery. Researches have shown that Mycobacterium tuberculosis (Mtb) encodes 20 CYP450 enzymes which play important roles in the synthesis and metabolism of lipid, cholesterol utilization, and the electron transport of respiratory chain in Mtb. With the critical roles within the organism as well as the protein structures of six Mtb CYP450 enzymes being clarified, some of them have been highlighted as potential anti-tuberculosis targets. In this paper, the phylogenetic analysis, the structural features, and the enzymatic functions of Mtb CYPs, as well as the mechanism of interactions with selective inhibitors such as azole antifungal agents for the CYPs have been reviewed and summarized. The druggability of the CYPs has also been analyzed for their further utility as targets in high throughput screening and rational design of more selective inhibitors.
Antitubercular Agents ; chemistry ; pharmacology ; Azoles ; chemistry ; pharmacology ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; pharmacology ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Drug Delivery Systems ; methods ; Drug Discovery ; Humans ; Mycobacterium tuberculosis ; drug effects ; enzymology ; genetics ; Phylogeny ; Tuberculosis ; drug therapy ; microbiology

OBJECTIVE: To test the technical parameters of GlobalFiler® PCR Amplification Kit for its application to forensic application value and to investigate the genetic polymorphisms. METHODS: The validation was conducted in sensitivity, mixed samples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The amplification and detection of the genomic DNA from 373 unrelated individuals from Beijing Han nationality were extracted by automation workstation. RESULTS: Global-Filer® PCR Amplification Kit was adaptive to some mixed, degraded and inhibited samples. The power of sensitivity and adaptability and peak height balance showed well. The distributions of genotype frequencies for 21 STR loci in the population were all in accordance with Hardy-Weinberg equilibrium (P > 0.05). The PIC value of the 21 STR loci was among 0.536 to 0.940; the H value was among 0.558 to 0.933; the DP value was among 0.783 to 0.992; the PE value was among 0.243 to 0.874. CONCLUSION GlobalFiler® PCR Amplification Kit is suitable for criminal cases and DNA database in forensic practice. And 21 STR loci in Beijing Han nationality have high polymorphism, which have application value in forensic practice and population genetics.
Asian People/genetics* ; Beijing ; Databases, Nucleic Acid ; Ethnicity ; Gene Frequency ; Genetic Loci/genetics* ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction/standards* ; Polymorphism, Genetic ; Reproducibility of Results ; Species Specificity

Objective To develop a rapid and efficient single cell PCR method for the non-invasive prenatal hemophilia A genetic diagnosis.Methods Six STR markers (DXS15,DXS9901,G6PD, DXS1073,DXS1108 and F8Civsl3) closely linked to F Ⅷ gene were examined in 118 healthy individuals and 12 hemophilia A families by muhi-plex fluorescent PCR.Three markers with the higher heterozygote rate and diagnostic rate were chosen and combined with amelogenin gene to establish the four-plex nested fluorescent PCR on single lymphocyte level.Results The single lymphocyte amplification rates of Amelogenin,DXS15,FSCivsl3 and DXSI073 were 94.3%,91.4%,100% and 100% in the normal male,and 100% ,97.1% ,97.1% and 97.1% in the normal female's respectively.The female was heterozygous in DXS1073 and DXS15.However,no allele dropout was found in both markers.Neither the false negative nor the false positive amplification was observed.Conclusion The single cell four-plex nested fluorescent PCR method established in this study is convenient and efficient,and will be hopefully employed in the clinical non-invasive prenatal genetic diagnosis for hemophilia A.

Objective To develop a liquid bandage for self aid of war wound.Methods Solution A and B were prepared containing functional polymers derived from the biocompatible poly (ethylene glycol) and hyaluronic acid,which had ability of fast crosslinking reaction when they were mixed.Among them,A solution mainly comprised of mercapto group-containing hyaluronic acid derivative (HA-SH),water-solubility starch,propylene glycol,benzoic acid and etc.B solution contained pentaerythritol tetraacrylate derivative (4arm-PEG-Acrylate) and benzoic acid.The structure of mercapto group-containing hyaluronic acid derivative was confirmed by IR,1H-NMR and etc.HA-SH and 4arm-PEG-Acrylate were formulated to different concentrations (W/V) in different buffers.The two kinds of solution were mixed in different ratios,and in situ crosslinking hyaluronic acid hydrogel was obtained;the crosslinking time was recorded.The adhesive force and waterholding capacity of the hydrogel after crosslinking were measured by adding excipients such as water-solubility starch.Results In case the concentration of HA-SH and 4arm-PEG-Acrylate was 2％ and pH value was 11,the hydrogel film on wound skin would be quickly formed in 20 seconds for wound dressing after spraying A and B solution onto the wound skin.Conclusion The liquid bandage has strong adhesive force,high water-holding capacity and controllable crosslinking time,so it could be used as a novel wound dressing for war wound.

Objective To verify the clinical applicability of the published standard intervals of routine clinical chemistry (WS/T404 .1‐2012 ,WS/T404 .2‐2012) based on the health examination results of Han and Uygur populations in Urumqi .Methods This was a retrospective study .The results of serum TP ,ALB ,ALT ,AST ,ALP ,GGT from healthy examination individuals of Han and Uygur populations (from 2013 August to 2015 January) were collected and the healthy cases (age range:20 -79 years old) were chosen to calculate the 2 .5% and 97 .5% percentiles ,excluding the significant abnormal results according to the Medical Deciding Level 2 recommended by Staland .The percents of health cases not falling in the published standard interval were calculated to meet the judgment criterion of verification (<10% ) .Results The test of normality revealed that the Han and Uygur's results of all veri‐fied items were skewed distributions .The 2 .5% and 97 .5% percentiles of the results of two populations were as follows ,TP(Han 65 -81 g/L ;Uygur 64-81 g/L) ,ALB(Han 41-53 g/L ;Uygur 40-52 g/L) ,ALT(Han:male 9-51 U/L and female 7-42 U/L ;Uygur:male 9-53 U/L and female 6-43 U/L) ,AST(Han:male 14-42 U/L and female 12-37 U/L ,Uygur:male 12-42 U/L and female 12-38 U/L) ,ALP(Han:male 45-119 U/L ;Uygur:male 47-122 U/L) ,ALP(female 20-49 years old:Han 35-95 U/L and Uygur 40-104 U/L) ,ALP(female 50-79 years old:Han 43-131 U/L and Uygur 51-132 U/L) ,GGT(Han:male 11-71 U/L and female 8-54 U/L ;Uygur :male 11 -73 U/L and female 7 -55 U/L ) .The percents of AST results for Han's male , Uygur's male and Uygur's female not falling in the published standard reference interval were slightly over 10% ,but AST results o‐ver 10% were mainly under the lower limit of the published standard reference interval .The health case percents for the other veri‐fied items of Han and Uygur populations not falling in the published standard reference interval were under 10% .Conclusion The published standard reference intervals of routine clinical chemistry (WS/T404 .1‐2012 ,WS/T404 .2‐2012) are applicable in our la‐boratory for the detection of Han and Uygur population .

Objective To test the technical param eters of GlobalFiler?PC R A m plification K it for its ap-plication to forensic application value and to investigate the genetic polym orphism s. Methods The valida-tion w as conducted in sensitivity, m ixed sam ples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The am plification and detection of the genom ic D N A from 373 unre-lated individuals from B eijing H an nationality w ere extracted by autom ation w orkstation. Results Global-Filer?PC R A m plification K it w as adaptive to som e m ixed, degraded and inhibited sam ples. The pow er of sensitivity and adaptability and peak height balance show ed w ell. The distributions of genotype fre-quencies for 21 STR loci in the population w ere all in accordance w ith H ardy-W einberg equilibrium (P>0.05). The PIC value of the 21 STR loci w as am ong 0.536 to 0.940; the H value w as am ong 0.558 to 0.933; the D P value w as am ong 0.783 to 0.992; the PE value w as am ong 0.243 to 0.874. Conclusion GlobalFiler?PC R A m plification K it is suitable for crim inal cases and D N A database in forensic practice. A nd 21 STR loci in B eijing H an nationality have high polym orphism , w hich have ap-plication value in forensic practice and population genetics.

Objective To identify Anaplasma species circulating among livestock and rodents from Xitianmu Mountain area in Zhejiang province , Southeastern China and to analyze variations regarding to their 16S rRNA gene.Methods Samples of spleen, liver and blood were collected to extract DNAs .The 16S rRNA gene fragments of Anaplasma species were amplified by using a nested PCR and then sequenced .Ho-mology analysis was conducted by using BLAST program .The multiple sequence alignment and phylogenetic analyses comparing with the sequences of other Anaplasma species in GenBank were conducted by using MEGA 5.0 software.Results The 16S rRNA gene fragments of Anaplasma were detected in 1 cattle, 8 goats, 5 Rattus confucianus, 1 Apodemus agrarius, 1 Berylmys bowersi and 1 squirrel out of 129 animals. The natural infection rate of Anaplasma was 13.2% in animals from Xitianmu Mountain area in Zhejiang . The alignment and phylogenetic analyses indicated that there were at least four Anaplasma species prevalent in livestock and rodents from Xitianmu Mountain area , including Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma centrale and Anaplasma bovis.Moreover, there was a variant that obviously differed from Anaplasmma bovis and other Anaplasma sp.in GenBank.Conclusion The Anaplasma infection was detected among livestock and rodents from Xitianmu Mountain area in Zhejiang province .A newly discovered variant in rodents was likely to be a novel species .More close attention should be paid to Anaplasma infec-tion among human in Xitianmu Mountain area .

Objective To investigate the synergistic effect of 11 β-hydroxysteriod dehydrogenase (11 β-HSD1) and S100A16 on the differentiation of3T3-L1 preadipocytes and its mechanism.Methods Lentiviral vectors PLJM1-11β-HSD1 and PLJM1-S100A16-GFP were respectively constructed and co-transfected into 3T3-L1 preadipocytes.The cell strains expressing 11 β-HSD1/S100A16 were screened with 2.5 μg/ml puromycin for two weeks.Western blot was employed to verify the lentiviral carrier transfection effects.The expressions of marker genes related to the adipocyte differentiation were detected by mean of realtime PCR.Oil red O staining was used to observe the lipid droplet accumulation and the content of triglyceride was measured after differentiation of preadipocytes.The effect of 11β-HSD1 and S100A16 on PPARγ promoter activity was detected by luciferase reporter gene.Results Compared with the empty vector group,the expressions of 11β-HSD1 and S100A16 protein in the lentivirus cotransfected 3T3-L1 cell strain were significantly higher.After 3T3-L1 cell strain co-expressing 1 1β-HSD1 and S100A16 was induced to differentiate for 8 days,the lipid droplets accumulation and triglyceride content were siginificantly increased,along with increased expressions of adipocyte differentiation marker genes such as PPARγ,CCAAT/enhancer binding protein α,lipoprotein lipase,fatty acid synthase,and adipocyte fatty acid-binding protein,in comparison with 11 β-HSD1 or S100A16 overexpression.The result of reporter gene indicated that 11 β-HSD1/ S100A16 enhanced PPARγ promoter activity.Conclusions 11β-HSD1 and S100A16 may jointly promote the differentiation of 3T3-L1 preadipocytes through a synergistic effect on PPARγexpression and play a critical role in the development of obesity.

This study was to build a canine portal hypertension model by intra-portal administration of high polymer material polyurethane and organic solvent tetrahydrofuran mixed solutions in order to evaluate the effectiveness of the model. Twelve local crossbreed dogs were selected randomly, with intra-portal administration of 8% (weight/volume) polyurethane- tetrahydrofuran solutions through an incision in the upper abdomen to build the portal hypertension model. We measured the portal vein pressure before modeling, during modeling, and four-, eight-, and twelve- weeks after modeling, respectively. Then we evaluated the effectiveness of the model comparing values of data with those data obtained before modeling started, which were regarded as the normal values. The results showed that the portal vein pressure rose by 2. 5 times after the solution administrated instantly as much as that before modeling, and maintained at 1. 5 times after 4 weeks. This method presents an easy operation, low animal mortality and reliable model of portal hypertension. Its less abdominal adhesions and its ability in keeping normal anatomic structure specially make it suit for surgical research of portal hypertension.
Animals ; Disease Models, Animal ; Dogs ; Furans ; adverse effects ; Hypertension, Portal ; Polyurethanes ; adverse effects ; Portal Vein ; physiopathology