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

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.

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.

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

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

Objective To understand the association of human leukocyte antigen (HLA)-DRB polymorphism and patients diagnosed as hemorrhagic fever with renal syndrome (HFRS). Methods HLA-DR allele polymorphism was detected by PCR-sequence specific primers (PCR-SSP). Hantavirus (HV) typed as Hantaan virus (HTNV) and Seoul virus (SEOV) in patients were detected by RT-heminested PCR. Results The gene frequency of DRB1*0401-0411, *1001 and *1101-1105 in HFRS case group were 3.1%, 2.2% and 15.7% respectively. Compared with control group, it was significant higher in HFRS case group (RR=13.87, 9.72 and 2.00 respectively with Chi-square value as 10.006,6.324 and 6.472 respectively, P<0.05). When compared with HFRS case group, the gene frequency of DRB1*1501-1502, DRB4 and DRB5 in control group were 11.0%, 19.0% and 16.9% respectively, markedly lower than in patients (RR=0.45, 0.58 and 0.23 respectively. Chi-square values were 6.138, 4.583 and 21.076 respectively, P<0.05). There was no significant difference in other HLA-DR gene frequencies. Mixed infection was found in Hubei, with HTNV slightly more than SEOV. Distinct hantaviruses could coexist in either different or the same geographic or ecological zores in Hubei province. Patients with HLA-DRB1*1101-1105 alleles were 81.8%(27/33) infected by HTNV and only 18.2% infected by SEOV, which had significant difference (P<0.05). Conclusion DRB1*0401-0411,*1001 and *1101-1105 were possibly associated with increased susceptibility to HV infection. On the other hand there was an inverse correlation among HFRS, DRB1*1501-1502, DRB4 and DRB5.

Objective To assess the new edition of WHO Japanese Encephalitis (JE) Surveillance Standards (WHO Standards) based on syndrome surveillance data and to provide field evidence regarding the standards. Methods Based on syndrome surveillance data, acute encephalitis syndrome (AES) case was categorized, according to the WHO Standards. A cohort study was applied to estimate the AES definition set in the Standard and relative risk was computed to estimate the existence and intensity of statistical correlation between AES and JE cases. Percentage of attributable risk was counted to describe the coverage of AES for JE cases in the studied population. Sensitivity,specificity, Youden index and positive predictive value of AES components were calculated for the purpose of identifying the clinical values under the screening program. Results 1424 suspected cases were evaluated in the surveillance program and 1396 cases with ELISA result, of which 109 positive cases were detected. According to the "standardized" classification, a total of 706 cases in line with AES case deftuition, were categorized into 83 cases of JE, 425 cases of AES unknown and 198 cases of AES other agent. In the cohort study,a relative risk of 4.62 (95% CI:2.80-7.63 ) and the percentage of attributable risk as 78.35% (95% CI: 64.25% -86.89% ) were observed. Conclusion The AES definition for JE was significantly effecting on the screening programs and a strong correlation strength was observed in the study. AES syndrome could cover most of the JE cases. "Convulsions",with appreciative screening value, was recommended to be involved into the new version of the WHO Standards.

Objective To investigate the antibacterial molecular mechanism of Traditional Chinese Medicine Coptis rhizome against Yersinia pestis(Y.pestis).Methods The method based on whole genome DNA micrnarray of Y.pestis was used.The minimal inhibition concentration(MIC)of berberine to Y.pestis was determined with liquid dilution method.Then gene expression profile of Y.pestis was performed after exposed to berberine at the concentration of 10×MIC for 30 minutes.Total RNA extracted and purified from Y.pestis and reverse-transcribed to cDNA,then labeled by Cy-dye.Finally,the labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software.Results The gene expression profile data revealed that the response of Y.pestis to berberine was a global phenomenon.A total of 360 genes changed significantly.Among them,333 genes were up-regulated,27 down-regulated.These differentially expressed genes were further classified into 24 different functional categories based on the genomie annotation of Y.pestis CO92,in which the number of mainly related genes were 83,75 and 48,including cell envelop,unkown,transport/binding proteins functions.The 40 genes related to the metabolism were upregulated,which was a remarkable change.Conclusion Our results have revealed the general gene expression changes of Y.pestis in response to berberine and demonstrated the antibacterial molecular mechanism of the Coptis rhizome.The major mechanism of Y.pestis in response to berberine is the upregulation of genes related to the metabolism.