Objective To investigate the mechanisms for cytopathic effect(CPE) of wide-type rubella virus(RV)in ECV304 endothelial-like cells.Method ECV304 cells were cultured in MEM supplemented with 10% fetal calf serum,glutamine,sodium bicarbonate,penicillin and streptomycin at 37℃ with 5% CO-2 in a humidified incubator,and then grown in maintenance medium with 2% fetal calf serum to avoid overstimulating cell growth.Infected cells and control cells were harvested at day 2,4, and 6 after infection for following experiments.RT-PCR and indirect immunofluorescence were used to detect RV infection.Phase-contrast and electron microscopy were performed to analyze morphology changes in infected and uninfected groups.TUNEL assay was carried out to study wide-type RV-induced apoptosis.Result In RV-infected ECV304 cells,cytopathic effects were first observed approximately at 2-3 days post-infection.The majority of cells with CPE changes were of detachment from the monolayer,rounding of cells,cell shrinkage and membrane blebbing were seen.At 4-6 days post-infection,the detached cells increased clearly;the CPE regions on the monolayer were extended,and the infected cells in the CPE regions showed in spindly form with wide spacing.RT-PCR and indirect immunofluorescence results indicated RV infection in ECV304 endothelial-like cells.Using electron microscopy,RV particles and its cytopathic effects including the condensed chromatin,fragmented nuclei and clustering of mitochondria with dramatic changes around nuclear were further examined.TUNEL results showed that the difference of apoptosis index between the infected group and uninfected group was significant(P

Objective: To investigate the relationship between postprandial glucose and cardiovascular disease.Methods: Ninety-four patients were divided into an IFG,an IGT and a DM group,and observed for the incidence of cardiovascular disease and the risk indexes of cardiovascular disease,such as CIMT,TC,TG,HDL-C,LDL-C,BMI,SBP,DBP and CRP.The correlation between postprandial glucose and the risk indexes were analyzed. Results: The risk indexes were obviously higher in the IGT and DM groups than in the IFG group(P0.01).Postprandial glucose was correlated positively with the risk indexes,but negatively with HDL-C.Conclusion: Postprandial hyperglycemia increases the incidence of cardiovascular disease.

Objective To study the effect of small interfering RNA(siRNA) of human papillomavirus(HPV) 18(E6) gene on apoptosis of HPV-related cervical HeLa cell line.Methods siRNA targeting HPV18 E6 mRNA was designed and generated by PCR amplification.The PCR products containing U6 promoter and the siRNA sequence were then transfected into HeLa cells via Lipofectamine()~(TM)2000.Cell viability was determined by MTT assay.Apoptosis was detected by morphological observation and flow cytometry analysis.The expression level of HPV18 E6 mRNA was assayed by RT-PCR.Results The cell growth and viability of(siRNA) transfected group were significantly inhibited(P

A questionaire before class,an analysis and an interview after exam were applied to evaluate the effect of bilingual teaching in biochemistry to provide important experiences and directions for carrying out bibingual teaching in the future.

Objective To prepare 60 mer oligo microarray chips for detecting human immunodeficiency virus (HIV) and for the clinical application in the detection of AIDS patient. Methods Oligonucleotide probes were designed according to the sequence information of two types of HIV. Oligo microarray was prepared by using Cartesian Microarrayer. Products of the restrictive display PCR were labeled with Cy3. Furthermore, the PCR products were sequenced. Results Using the oligo microarray, HIV infection could be detected in laboratory as well as in clinical assays. Results of hybridization indicated that 1 AIDS patient was positive and 20 health people were negative. The results obtained by sequencing confirmed the results obtained by oligo microarray studies. Conclusion The HIV 60 mer oligo microarray could be used in detecting patient HIV infection and analyzing its genotypes.

Objective To design a microarray of ~60mer oligonucleotide for detection and sub-typing of human papillomavirus (HPV). Methods The type-specific oligonucleotide probes of 4 different types of HPV (6, 11, 16, 18) were designed by using biological software Arraydesigner 2.0, which analyzed the whole genome sequences of HPV and selected optimal probes with high specificity, identical length and similar melting temperature (Tm). These probes were synthesized and printed onto the surface of glass slides in order to prepare a low-density microarray. HPV samples were labeled with fluorescence dyes Cy3 using a method of restriction display PCR (RD-PCR). HPV plasmid DNA was restricted with Sau3AⅠ to produce multiple fragments which were ligated to adaptors subsequently and used as PCR template. PCR labeling was performed with the fluorescently labeled universal primer (Cy3-UP) whose sequence is designed according to the adaptor of the RD-PCR approaches. The labeled samples were then hybridized with the oligonucleotide microarray. Results Both single and multiple HPV DNA samples could be detected with oligonucleotides microarray, and the corresponding HPV subtypes were recognized as well. And no signals were detected in all the negative and blank control spots. Conclusion 60mer oligonucleotide microarray designed by appropriate bioinformatics software can be applied to HPV detection and genotyping on gene level.

Objective To evaluate the value of preoperative color Doppler flow imaging findings for predicting possible difficuulties encountered during laparoscopic cholecystectomy (LC).Methods Eighty-six Datients with acute cholecystitis underwent color Doppler flow imaging examination were divided into operation difficult group(67 cases)and operation easy group(19 cases)according to the diffculty score.The parameters were measured pre-operation including the volume of gallbladder,the thickness of the gallbladder wall,the condition of arterial flow in the gallbladder wall,the conditions of gallbladder cavity and gallbladder fossa and the intra-and extra-hepatic bile duet.The relationships among imaging results,operation difficulties and operation findings were investigated.Results Gallbladder volume,gallbladder wall thickness.the presence rates of plentiful arterial flow in the gallbladder wall,adhesion of gallbladder and stone incarceration In operation difficult group were significantly different from those in operation easy group [(52.6±14.6)mm~3 vs(32.6±10.4)mm~3,(9.7±4.1)mm vs(3.8±0.9)mm,89.5%(17/19)vs 17.9%(12/67),78.9%(15/19)vs 11.9%(8/67),10.5%(2,19)vs 0(0/67)](P<0.05 or<0.01).The accurate rate was 94.2%(81/86)in predicting.Conclusion Preoperative color Doppler flow imaging is helpful in predicting difficulties of LC.

Objective To explore the effect of the mutant and expression level of N-ras on chronic myelogenous leukemia. Method We investigated the mutant by direct sequencing in a K562 human chronic myelogenous leukemia cell line, with determination of the expression level of N-ras mRNA in K562 by RT-PCR. Result No single point mutation was detected in K562 cell line, furthermore, the expression level of N-ras gene is abnormaly high in contrast to normal human. Conclusion Our results indicated that the expression level of N-ras gene was obviously high in K562 cell line, and the underlying mechanism was not only mutation, so that further investigation is called for.

Objective To study the double restriction fluorescent labeling (DRFL) method for fluorescent labeling of trace DNA samples and its effect in enhancing the pathogen detection sensitivity of microarray assays. Method SARS-CoV RNA samples were reversely transcribed and then further amplified with the restriction display (RD)-PCR and fluorescently labeled by conventional restriction labeling directly with Cy-universal primer and the novel double labeling with Cy-universal primer and CydNTP. The labeled samples were applied to the microarray with the viral probes, processed and analyzed. Results Compared with the conventional method, DRFL labeling resulted in 3. 5835 times higher fluorescent intensity of all the SARS probes on average, even though increased fluorescent intensities for different probes varied considerably. Conclusion Signal to noise ratio can be enhanced by the DRFL method which improves the sensitivity of microarray technology in trace pathogen detections.

According to the teaching practice in biochemistry laboratory course,we describe the characteristics of international students'teaching,teaching preparation,teaching course and so on.These experiences may provide an important source of information for teaching practice in the future.