BACKGROUND:Induction of osteoblasts differentiating into osteocytes is a hot spot in tissue engineering;however, the regulatory mechanism underlying differentiation has not been ful y elucidated. MicroRNA, as an endogenous smal RNA molecule, can regulate post-transcriptional gene expression by binding to the 3’ nontranslated region of the target gene mRNA, which also has been found to play an important regulatory role in osteocyte differentiation. OBJECTIVE:To study the regulation of miR-155 on osteoblast differentiation and the underlying mechanism. METHODS:The mouse osteoblast cel lines MC3T3-E1 were selected and induced by mouse bone morphogenetic protein-2 (BMP2, 200 ng/mL) and then the miR-155 mRNA expression was determined by quantitative real-time PCR at 1, 3, 7 and 14 days. MC3T3-E1 cel s were divided into control, BMP2, miR-155 and miR-155 inhibitor groups, fol owed by cultured withα-MEM medium, BMP2, miR-155 and miR-155 inhibitor, respectively, for 2 weeks. RESULTS AND CONCLUSION:After induction using BMP2, miR-155 expression was downregulated in a time dependent manner. The staining intensity of alizarin red in the BMP2 group was significantly higher than that of the control group, and the activity of alkaline phosphatase and mRNA expression were also significantly higher than those in the control group (P<0.01). The staining intensity of alizarin red, activity of alkaline phosphatase and mRNA expression in the miR-155 group were significantly lower than those in the control group (P<0.01), while al above measurements were reversed significantly by miR-155 inhibitor (P<0.05). miR-155 could bind to the 3’ untranslated region of SMAD5 mRNA and significantly downregulated the expressions of SMAD5 protein and mRNA in MC3T3-E1 cel s (P<0.01). These results show that miR-155 can inhibit MC3T3-E1osteogenic differentiation by downregulating SMAD5 expression.

Objective To investigate the prevalence and risk factors of metabolic syndrome (MS) in residents in Pudong New District of Shanghai. MethodsA total of 5 584 residents aged 20-80 years were randomly selected from Pudong New District of Shanghai through multistage sampling and interviewed from April to July of 2008. Metabolic syndrome was defined according to three diagnostic criteria for MS, issued by the modified National Cholesterol Education Program Adult Treatment Panel Ⅲ criteria ( NCEP-ATP Ⅲ ), International Diabetes Federation (IDF), and Chinese Diabetes Society (CDS). ResultsThe crude prevalences of MS in the adult population in Pudong New District were 18.2％ and 13.1％ standardized ( male 19. 1％, female 17.4％, the age-standardized 15.6％ and 13.2％ ) with CDS criterion, 31.8％ and 24.4％ standardized ( male 28.4％ ,female 35.1％ ,the agestandardized 22. 7％ and 25.0％ ) with NCEP-ATP Ⅲ criterion, and 21.7％ and 17.0％ standardized ( male 15.9％ ,female 26.7％, the age-standardized 13.8％ and 19.2％ ) with IDF criterion. The age-specific prevalence of MS increased according to three diagnostic criteria, and the age-adjusted prevalence was higher in males than females in junior age groups and higher in females than males in senior ones. Significant differences were present among region, education, marriage status, smoking, work intensity, recreation, and physical activity according to some diagnostic criteria. ConclusionsSubstantial proportions of adults in Pudong New District of Shanghai suffer from metabolic syndrome, and there exists a tendency for young people involved. MS has become a noteworthy public health problem. It suggests that community-integrated control strategy of MS should be made a priority.

Adaptor Proteins, Signal Transducing ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Proteins ; metabolism ; RNA, Messenger ; genetics

To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

0bjective To determine the optimal contrast protocols for 64.slice spiral CT coronary angiography in order to reduce the volume of contrast injection.Methods One hundred fifty pailents scheduled to undergo 64-slice spiral CT coronary angiography were prospectively randomized into the following five groups with different injection protocols:group l:uniphasic injection without a flush;group 2:biphasic injection with a flush;group 3,group 4 and group 5:triphasic injection with a diluted contrast material with 3:7.5:5.7:3 contrast/saline ratio respectively.Attenuation was measured in the right atrium,right ventricle,left atrium,left ventricle,ascending aorta,fight coronary artery and left coronary artery and analyzed with single factor variance test(ANOVA).The quality the coronary artery images was evaluated and compared using person Chi-Square.Results The total contrast material vohme were (67.0±5.3)ml,(59.9±4.9)ml,(62.9±3.2)ml,(69.2 4±5.7)ml and(70.9 4-4.6)ml in five groups respectively(F=27.43,P<0.01).Image quali～scores of coronary arteries were significant different among five groups(X2=18.81,P<0.05).There were signiflcandy differences in artifacts of the superior vena cava among five groups(X2=31.44,P<0.01).The artifacts in the superior vena cava in group 1 was the most,and in group 2 was the least.The mean enhancement values of right and left coronary arteries in group 2 were significantly greater than those in other groups(F=2.47 and 4.10,P<0.05).The visualization of both left ventricle and right ventricle cavities W88 the best in group 3.Conclusion Biphasic injection and triphasic injection are better than uniphasic injection for 64-slice spiral CT coronary angiography and triphasic injecfion is better than biphasic injection for the visualization of both left ventricle and right ventricle cavities.

Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

To study the relationship between the interleukin (IL)6 -572G/C polymorphism and risk of hepatocellular carcinoma (HCC) in men.A hospital-based case-control study was conducted with 500 male HCC patients without tumor history in other organs and 590 healthy male controls without history of tumors or chronic diseases. All HCC cases were diagnosed by histopathology. The controls were recruited from the Department of Orthopedic Trauma and Ophthalmology at the same hospital. The IL-6 promoter -572G/C polymorphism and its genotype variants were detected by real-time fluorescence quantitative PCR. The Chi-squared test and unconditional logistic regression analyses were applied to determine the risk of HCC among men carrying the different genotype variants.The frequencies of alleles and distribution of genotypes in the -572G/C loci were not significantly different between the HCC cases and controls (P more than 0.05). The Chi-squared test indicated that the polymorphisms of the loci were not associated with HCC in our male population. However, after adjusting by multivariate logistic regression, the odds ratio (OR) of HCC for the G allele (CG + GG genotypes) carriers was 1.31 (95% confidence interval (CI): 1.00 - 1.71) compared with the CC genotype. Among the male HBV carriers, the CG genotype increased HCC risk significantly (OR = 1.60, 95% CI: 1.14 - 2.24) compared with the CC genotype. A trend test indicated that HCC risk was significantly increased with the numbers of G alleles (P trend less than 0.05). Breslow-Day tests of homogeneity of the ORs indicated an interaction between hepatitis B virus (HBV) infection and polymorphisms of IL-6 (P less than 0.05). The synthetic odds ratio (OReg) of HBV infection and harboring a G allele was 5.95 (95% CI: 3.99-8.87), which represented a super multiplication interaction.Polymorphism of the IL-6 promoter -572 loci may be associated with HCC occurrence in men. Moreover, there is a super multiplication interaction for HCC risk between HBV infection and harboring the IL-6 G allele.
Adult ; Alleles ; Carcinoma, Hepatocellular ; genetics ; pathology ; Case-Control Studies ; Genotype ; Humans ; Interleukin-6 ; genetics ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Risk Factors

BACKGROUNDStreptococcus mutans (S. mutans) is the prime pathogen of dental caries. There are few reports that studied the relationship between S. mutans, bacteria and dental caries in permanent teeth when compared to those in primary teeth. This study aimed to detect S. mutans and bacteria of dental caries and non-caries groups in permanent teeth from a north China population by real-time polymerase chain reaction (PCR) and compare the relationship between the number of these bacteria and the prevalence of dental caries in permanent teeth.

METHODSHuman saliva samples were collected from 142 subjects with permanent teeth. According to their dental tooth (DT), 142 subjects were divided into a dental caries group (DT ≥ 1) and a non-caries group (DT = 0). With specific primers for S. mutans and 16S rRNA, the total number of S. mutans and total bacteria of 142 saliva samples were detected by real-time PCR and statistically analyzed.

RESULTSThere was no significant difference between the detection rates of S. mutans (P = 0.118) and medians of S. mutans (P = 0.115). The ratio of S. mutans to total bacteria in people with dental caries was significantly higher than in those without caries (P < 0.001), but the total number of bacteria in people with dental caries was significantly lower than in those without caries (P < 0.001).

CONCLUSIONSS. mutans had different effects on caries in the permanent teeth of several individuals from a north China population. The ratios of S. mutans to total bacteria in saliva detected by real-time PCR with Sm479F/R and 16S RNA primers were closely associated with the prevalence of dental caries in the same population. These assays may be useful for the assessment of an individual's risk of dental caries.

Adolescent ; Adult ; Aged ; Bacteria ; isolation & purification ; Dental Caries ; microbiology ; Female ; Humans ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Saliva ; microbiology ; Sensitivity and Specificity ; Streptococcus mutans ; isolation & purification ; Tooth ; microbiology

To investigate molecular mechanism of traditional Chinese medicine Rheum offcinale against Yersinia pestis, whole genome DNA microarray that contains 4005 annotated genes of Y. pestis was used. The minimal inhibition concentration (MIC) of R. offcinale extract against Y. pestis was determined by liquid dilution method. The gene expression profile of Y. pestis was performed after exposured to R. offcinale extract at a concentration of 10 X MIC for 30 and 60 minutes. The total RNA extracted and purified from Y. pestis were reverse-transcribed to cDNA and labeled by Cy3-Cy5 dye. The labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software. The microarray data was confirmed by RT-PCR. The platform of the DNA microarray-based bacteria transcriptional profiling was eshtablished. The results revealed general gene expression changes of Y. pestis were a global phenomenon. Down-regulation of genes encoding proteins involved in ribosome protein synthesis was a remarkable change. Genes encoding cell envelope and transport/binding proteins were the major changed genes of the Y. pestis in response to R. offcinale.
Bacterial Proteins ; genetics ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; Microbial Sensitivity Tests ; Oligonucleotide Array Sequence Analysis ; RNA, Bacterial ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Rheum ; chemistry ; genetics ; Yersinia pestis ; drug effects

OBJECTIVETo determine the affinity of new opioid receptor ligands to cloned mu opioid receptors stably expressed in CHO cell.

METHODSThe binding characteristics of the opioid ligand [3H] diprenorphine (3H-dip) were studied by cellular biological techniques and radioligands binding in cloned mu opioid receptors stably expressed in CHO cells in saturation binding experiments, and were followed by competition binding experiments with a variety of new synthesized opioid receptor ligands.

RESULTSThe Kd and Bmax of [3H] diprenorphine bound to mu receptors were 1.06 nmol/L and 930 fmol/mg protein, respectively. Competition binding experiments revealed that ligand 3# and 12# displayed much higher affinity than DAMGO and Morphine for the cloned mu opioid receptor. However, the affinities of ligands 2#, 6#, 8# and 9# were lower than DAMGO and Morphine.

CONCLUSIONThe present results suggest that the new ligands 3# and 12# have higher affinity to mu opioid receptors. However, ligands 2#, 6#, 8# and 9# have lower affinity to mu opioid receptors.

Animals ; Binding Sites ; Binding, Competitive ; CHO Cells ; metabolism ; Cloning, Molecular ; Cricetinae ; Diprenorphine ; pharmacology ; Ligands ; Receptors, Opioid, mu ; biosynthesis ; genetics ; metabolism