OBJECTIVE:To establish a method for the contents determination of astragalosidesⅠ,Ⅱ,Ⅲ,Ⅳ,lobetyolin and for-mononetin in Shenqi fuzheng injection. METHODS:HPLC was performed. The detector was evaporative light scattering detector, column was Ultimate XB-C18 with mobile phase of acetonitrile-0.5% formic acid (gradient elution) at a flow rate of 1.2 ml/min, column temperature was 30 ℃,and injection volume was 10 μl. RESULTS:The linear range was 0.62-5.67 μg for astragalosidesⅠ(r=0.999 6),0.78-6.31 μg for astragalosidesⅡ(r=0.999 6),0.36-3.48 μg for astragalosidesⅢ(r=0.999 7),0.81-6.72 μg for as-tragalosides Ⅳ(r=0.999 5),0.82-7.03 μg for lobetyolin(r=0.999 8)and 0.58-6.62 μg(r=0.999 7)formononetin;limit of quan-titation was 1.21,0.15,0.12,0.03,0.12,0.17 μg/ml;limit of detection was 0.35,0.35,0.04,0.01,0.03,0.04 μg/ml;RSDs of pre-cision,stability and reproducibility tests were lower than 2%;recoveries were 95.1%-101.1%(RSD=2.0%,n=9),95.2%-100.7%(RSD=1.9%,n=9),95.8%-100.2%(RSD=1.4%,n=9),96.2%-100.6%(RSD=1.7%,n=9),96.6%-101.2%(RSD=1.4%, n=9) and 95.9%-99.5%(RSD=1.2%,n=9),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the simultaneous determination of astragalosidesⅠ,Ⅱ,Ⅲ,Ⅳ,lobetyolin and formononetin in Shenqi fuzheng injection.

There is an increasing interest in single nucleotide polymorphism (SNP) typing in the forensic field.SNPs are very useful for deftning Y chromosome or mtDNA haplotypes and DNA phenotyping.We focus on comparative advantages of SNP typing over length variations and expected number of loci required to gain probabilities equal to sTR loci in use.This review also offers to the reader a state of the art of SNP genotyping technologies with the advantages and disadvantages of the different techniques and platforms for different forensic requirements.

Objective To investigate lead pollution of tap in market. Methods 16 taps (2 PPR taps and 14 metal taps)were collected and were sunk in distilled water for 4 hours,8 hours and 12 hours. Lead levels in the distilled waters were determined by atomic absorption spectrometry. Results No lead was detected for PPR taps. Of 14 metal taps,only one tap was not observed lead pollution. The top lead level was 56.54 ?g/L,the median was 5.79 ?g/L. These brass taps resulted in higher lead pollution than those taps with ceramic preformed core (P0.05). 72.2% of the samples with brass preformed core exceeded 10 ?g/L and only 9.5% for those taps with ceramic preformed core. These taps with registered trade mark had a slighter lead contamination than those without registered trade mark(P

Objective Genotype data of nine CODIS STR loci were gathered to examine the features of population differentiation and gene flow of seven Xinjiang minorities.Methods Heterozygosity,Nei's coefficient of genetic differentiation,Nei's genetic distance and Wright's F-statistics were calculated. Statistical tests using exact method were performed to measure the level of differentiation.Phylogenetic trees were constructed by Mega;AMOVA was processed by Arlequin.R-matrix model had been applied to describe the patterns of gene flow.Results It shows that average genetic heterogeneity for each population was above 0.7 with genetic differentiation coefficient below 2%.Statistical tests for population differentiation were significant for most of the loci.Phylogenetic analysis and AMOVA showed that all populations were divided into three main groups.The R-matrix analysis reflected that Uygur,Kirgiz and Ozbek had more amounts of gene flow than other populations,while the pattern of Hui was more isolated.Conclusion The seven minorities in Xinjiang are independent populations,while the level of differentiation is at average.The relationship in evolution is not far from each other,with wide gene flow.

OBJECTIVE To type the genes of plasmid DNA in 54 clinical Klebsiella pneumoniae isolates producing extended -spectrum beta-lactamases (SHV) by denaturing high-performance liquid chromatography (DHPLc) and evaluate their sensitivity and specificity, and explore a rapid and convenient method for detecting the antibiotic resistance of K. pneumoniae. METHODS Plasmid DNA from each extended-spectrum beta-lactamase (SHV) producing strain was subjected to PCR amplification. After we performed DNA sequencing of these amplicons and identification of mutation and their genotype, DHPLc was undertaken to investigate whether its results correlate the distinctive chromatogram with each genotype. RESULTS All the strains were found abnormal elution peaks (two or three peaks) which were different from each other. The result of DNA sequencing demonstrated that all the strains had DNA mutation in comparison with SHV-1. Moreover, DHPLc could produce specific peak patterns that correlate with genotype. CONCLUSIONS The sensitivity of DHPLc is 100% in this study. And each genotype is corresponded to specific peak pattern. So we can use DHPLc technique to type the genes of plasmid DNA in K. pneumoniae and detect mutations rapidly. DHPLc not only has high accuracy , but also is a convenient and rapid technique for the detection of mutation in the bacterial genome. It has a great potential clinical value.

Nine short tandem repeat (STR) markers (D3S1358, VWA, FGA, THO1, TPOX, CSFIPO, D5S818, D13S317, and D7S820) and a sex-identification marker (Amelogenin locus) were amplified with multiplex PCR and were genotyped with a four-color fluorescence method in samples from 174 unrelated Han individuals in North China. The allele frequencies, genotype frequencies, heterozygosity, probability of discrimination powers, probability of paternity exclusion and Hardy-Weinberg equilibrium expectations were determined. The results demonstrated that the genotypes at all these STR loci in Han population conform to Hardy-Weinberg equilibrium expectations. The combined discrimination power (DP) was 1.05 x 10(-10) within nine STR loci analyzed and the probability of paternity exclusion (EPP) was 0.9998. The results indicate that these nine STR loci and the Amelogenin locus are useful markers for human identification, paternity and maternity testing and sex determination in forensic sciences.
Amelogenin ; China ; Dental Enamel Proteins ; genetics ; Electrophoresis ; Ethnic Groups ; genetics ; Forensic Medicine ; methods ; Gene Frequency ; Genetics, Population ; Genotype ; Heterozygote ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sex Determination Analysis ; methods ; Tandem Repeat Sequences ; genetics

BACKGROUND:Previous studies have shown that minichromosome maintenance 3 is related with fluorosis, but the expression of minichromosome maintenance 3 in fluorosis patients is not clear yet.
OBJECTIVE:To analyze the mRNA expression level of minichromosome maintenance 3 in peripheral blood from patients exposed to fluoride and normal controls.
METHODS:Eleven patients with mild fluorosis by drinking water (exposure group) and 11 cases of control (non-exposure group) were selected for research. SYBRGreen1 real-time quantitative PCR was used to determine the mRNA expression of minichromosome maintenance 3 in peripheral blood mononuclear cel s, and the liver and renal function indicators were detected.
RESULTS AND CONCLUSION:mRNA expressions of minichromosome maintenance 3 in the exposure group and non-exposure group were (0.573 60±0.102 59) and (0.550 0±0.171 81), respectively, and there was no significant difference between two groups (P>0.05). There were no significant differences in the liver and renal function indicators between two groups. The results indicate that mild fluorosis has no significant effect on mRNA expression of minichromosome maintenance 3 in the peripheral blood mononuclear cel s. More indicators are needed to compressively analyze the effect of fluoride on the liver and renal functions.

Objective To probe the association between possible single nucleotide polymorphism (SNP) in the FGB promoter region and idiopathic deep venous thrombosis.Methods A prospective analysis was performed in both IDVT group and control group (120 cases each) followed by a duplex examination using gene sequencing technique and restriction fragment length polymorphism (RFLP) in the promoter region of fibrinogen gene β.Possible SNPs in this region were detected arranged before HardyWeinberg equilibrium test and Linkage disequilibrium (LD) analyses.Ultimately,we compared the genotype frequencies between the two groups and undertook a multiple Logistic regression.Results Six kinds of SNPs were determined in the promoter region of β-fibrinogen gene:-148C/T,-249C/T,-455G/A,-854G/A,-993C/T and-1420G/A.A stronger linkage disequilibrium was confirmed between-993C/T and -455G/A (r2 =0.699) ;-993C/T and-148C/T (r2 =0.509) ;-455G/A and-148C/T (r2 =0.556).Statistical differences of genotype frequencies between two groups were observed in-148C/T,-249C/T,-455G/A and-1420G/A polymorphisms (all P ＜ 0.05).Conclusions The risk of IDVT was 4.579 times higher with every 1 g/L increase of fibrinogen concentration.Allele-148T,-455G and-1420A are IDVT risk factors.-993C/T may indirectly affect IDVT through linkage disequilibrium with-455G/A and-148C/T.

Objective To investigate the antitumor activity of the procaspase-3 activator SM-1 in BGC-823 cells in vivo and in vitro and the mechanisms.Methods The inhibitory effects of SM-1 on proliferation of BGC-823 cells were evaluated using MTT method, the cell apoptosis rate was detected by flow cytometry, and the expression of caspase-3 protein and procaspase-3 mRNA was detected by Western blotting and RT-PCR, respectively.SM-1 Antitumor activity was evaluated using the xenograft of BGC-823 cells in nude mice.Results SM-1 effectively inhibited the proliferation in vitro and in-duced apoptosis of BGC-823 cells in a dose-dependent manner.After treatment with SM-1 for 48 h, the protein expression levels of caspase-3 and mRNA expression levels of procaspase-3 were increased.SM-1 significantly inhibited growth of BGC-823 xenograft tumor at the 300 mg/kg dose and the inhibition rate was 56.3%(P<0.05).Conclusion SM-1 can significantly inhibit the tumor growth of BGC-823 cells in vivo and in vitro.The mechanism is possibly related to the activation of procaspase-3 and induced apoptosis of tumor cells.

OBJECTIVE To understand characteristics of TEM-116 ?-lactamases through comparative study on resistance to antibiotics of clinical isolates of Klebsiella pneumoniae producing TEM-116 ?-lactamases.METHODS K.pneumoniae susceptibility to ?-lactamases was determined by disk diffusion tests,and their isoelectric points(PI) were detected using analytic isoelectric focusing(IEF),and resistance to antibiotics of clinical isolates of K.pneumoniae producing TEM-116 and TEM-1?-lactamases was studied.RESULTS Both of K.pneumoniae producing TEM-116 ?-lactamases and producing TEM-1 ?-lactamases were 100% resistant to AMP,and highly resistant to the first and second generation cephalosporin,but greatly susceptible to FEP and IPM.There was greatly difference between resistance to AMC,TZP,AMK,and GEN of clinical isolates of K.pneumoniae producing TEM-116 ?-lactamases and that of K.pneumoniae producing TEM-116 ?-lactamases,the TEM-116 isolates were higher resistant than TEM-1 isolates.Analytic IEF results showed that PI of TEM-116 ?-lactamases was 5.4,and most strains of K.pneumoniae TEM-116 ?-lactamases displayed two electrophoresis bands or more,only one strain of them just displayed one band,resistant to majority of antibiotics.CONCLUSIONS The results show that K.pneumoniae producing TEM-116 ?-lactamases are more resistant to antibiotics than K.pneumoniae producing TEM-1 ?-lactamases,and indicate TEM-116 ?-lactamases work as ESBLs.