Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Female ; Gene Frequency ; Humans ; Male ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

Objective To test the reliability and validity of the Chinese version of Profile of Fatigue Scale (PROF)among the patients with Sjogren′s syndrome. Methods The English version of PROF was translated into Chinese according to Brislin Translation Model. 107 patients with Sjogren′s syndrome were investigated to test its reliability and validity. Results The Cronbachαcoefficient , Guttman split-half coefficient, test-retest reliability and content validity of Chinese version of PROF were 0.976, 0.953, 0.872 and 0.875. Two factors were extracted by factor analysis and the cumulated variance was 80.75%. The structure of PROF was in line with the original scale. The criterion-related validity was 0.621 measured by comparing with visual analog scale of fatigue, and was -0.707 measured by comparing with vitality domain in the MOS item Short From Health Survey (SF-36). Conclusions The Chinese version of PROF has been proved to be reliable and valid. It can be used to measure the fatigue in Chinese patients with Sjogren′s syndrome.

This study was aimed to construct lentivirus-mediated shRNA expression vector targeting Bmi-1 and establish a stable cell line U266-li, so as to pave the way for further research on function of Bmi-1 and application of shRNA to gene therapy. One pair of oligonucleotide sequences targeted at human Bmi-1 mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pLVTHM vector. Virus particles were collected after the control or shRNA vectors were co-transfected with the psPAX2 packaging plasmid and the plasmid pMD2.G was enveloped into HEK-293T cells by using Lipofectamine2000. The U266 cells were transduced with 5 × 10(6) recombinant lentivirus-transducing units plus 6 µg/ml of polybrene. Real-time PCR and Western blot were used respectively to detect the expression of Bmi-1 and P14 after lentivirus transduction. DNA sequencing demonstrated that the lentivirus RNAi vector of Bmi-1 was constructed successfully and the virus was packaged in 293T cells. The titer of virus was 5 × 10(7) TU/ml. Stable transfected U266 cell line was established. As was expected, the mRNA and protein levels of Bmi-1 was reduced significantly in U266 cells after lentivirus transduction, whereas the mRNA and protein levels of P14 was upregulated. It is concluded that the lentiviral RNAi vector of Bmi-1 is constructed, and U266 stable cell line is established.
Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Polycomb Repressive Complex 1 ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics

This study was aimed to construct miRNA sponge targeting miR-20a and to establish a stable cell line Jurkat-S, paving the way for further research on function of miR-20a and application of RNAi in gene therapy. One pair of two-repeated oligonucleotide sequences containing bulged sites that are mispaired opposite miR-20a positions 9-12 was designed and synthesized with enzyme cutting sites. The annealed oligonucleotide fragments were subcloned into pCDNA3.0-L expressing vector. After double-enzyme cutting, the vector was ligated to the annealed oligonucleotide fragments again. Enzyme cutting and luciferase activity assay were performed for identification after four repeats. Then the ligated fragment was subcloned to lentivirus expressing vector. Virus particles were collected after the control or sponge vectors were co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pMD2.G into HEK-293T cells using Lipofectamine 2000. The Jurkat cells were transfused with recombinant lentivirus-transfusing units plus 6 µg/ml of Polybrene. Real-time PCR and Western blot were used to detect the mRNA and protein expression of P21 and E2F1 after lentivirus transfusion respectively. As a result, luciferase activity assay demonstrated that the sponge targeting miR-20a was constructed successfully and the virus was packaged in 293T. The titer of virus was 5×10(7) TU/ml. Stable transfected Jurkat-S cell line was established. As was expected, the mRNA and protein level of P21 and E2F1 was upregulated significantly in Jurkat-S cells. It is concluded that the miR-20a sponge is constructed successfully, and Jurkat-S stable cell line is established, in which the expression of miR-20a is inhibited stably.
Gene Expression ; Genetic Vectors ; Humans ; Jurkat Cells ; MicroRNAs ; genetics ; Plasmids ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate the effect of overexpression of S3 ribosomal protein (S3rp) gene on the resistance of leukemia cell to antitumor drugs.

METHODSBoth sense and antisense cDNA recombinants of S3rp gene were constructed with pcDNA3.1 expression vector. Subsequently, the sense S3rp cDNA recombinant was transfected into K562 cells while the antisense one into K562/DOX cells (a multidrug resistant cell line). In addition, empty pcDNA3.1 vector was transfected into the corresponding cells as negative controls. The chemosensitivity of cells was evaluated by MTT assay.

RESULTSSense S3rp cDNA transfected K562 cells were 5.8 times more resistant to doxorubicin than control cells did, whereas antisense S3rp cDNA transfected K562/DOX cells were 3.2 times less resistant to doxorubicin than control cells did.

CONCLUSIONOverexpression of S3rp gene plays an important role in the development of drug resistance in K562/DOX cells.

Antibiotics, Antineoplastic ; pharmacology ; DNA, Antisense ; genetics ; DNA, Complementary ; genetics ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; genetics ; Gene Expression ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Plasmids ; genetics ; Ribosomal Proteins ; biosynthesis ; genetics ; Transfection

Objective To assess the feasibility of three-dimensional contrast-enhanced ultrasonic cholangiography(3D-CEUSC) in cadaver liver.Methods The 3D-CEUSC was performed in 6 cases of cadaver liver.Image quality of 3D-CEUSC was evaluated.The visualization of branching orders,the degree of visibility and coincidence of morphous were compared with those of cholangiagraphy using fluoroscopy.Results The imaging quality of 3D-CEUSC was inferior to that of cholangiography with significant difference.The three-dimensional biliary tree structures were visualized in all 6 3D-CEUSC.The maximum visualization of branching orders in 3D-CEUSC was (3.67±0.52),which was equal to the results(4.00±0.63)by cholangiography (P=0.465).The degrees of visibility of biliary tree were equivalent with those by cholangiography in the first and second order with significant difference.The coincidence of morphous was excellent compared the images of 3D-CEUSC with direct X-ray cholangiography.Conclusion 3D-CEUSC is a new technique as a useful supplement to cholangiography in evaluation of biliary anatomy and variation before graft harvesting in LDLT.

Objective To investigate the relationship between the endoscopic characteristics of chronic gastritis,duodenitis,peptic ulcer and their histopathologic findings in children,and explore the relationship between Helicobacter pylori(Hp) infection and the severity of histopathologic changes of gastroduodenal mucosa in children. Methods Three hundreds and sixty-six children with chronic upper gastrointestinal symptoms who were examined by gastroscopy were enrolled.The gastric and duodenal mucosal biopsy specimens of all the patients were studied histopathologically. ResultsTypes of chronic gastroduodenal diseases in all these patients were: chronic gastritis(n=206,56.3%),chronic gastritis combined with duodenitis(n=112,30.6%),chronic gastritis combined with peptic ulcer(n=48,13.1%).There was chronic inflammation in gastric mucosa and duodenal bulb mucosa in all the cases examined by histopathologic examination.Hp infection was found in 106 cases(28.96%).The gastric mucosal inflammation was more severe in those with Hp infection than those without(P0.05).The were significant differences in the incidences of inflammation activity,atrophia and nodulus lymphaticus of gastric mucosa between those with Hp infection and those without(P

ObjectiveTo explore temperament characteristics of children with cerebral palsy.Methods364 children with cerebral palsy, ages from four months to seven years, were tested with modified Carey\'s Children Temperament Questionnaires. Temperament characteristics of children with different types of cerebral palsy, able and unable walk independently were compared.ResultsChildren with cerebral palsy had negative temperament including less active, low persistence, high distractibility and negative mood. Approach was different between children with diplegia and athtosis. There were no different temperaments among children with different lever of IQ. Children they could not walk independently had an obvious negative temperament.ConclusionChildren with cerebral palsy have significantly negative temperament.

Objective To investigate the characteristics of pulse wave velocity(PWV)and heart rate variability (HRV)and evaluate their feasibility in grading the cardiovascular risk in patients with isolated systolic hypertension (ISH).Methods Eighty-nine ISH patients and systolic-diastolic hypertension patients(DH,n=98)admitted in our hospital were submitted carotid-radial PWV(crPWV),carotid femoral PWV(cfPWV)and HRV,ISH patients were categorized depending on their risk grade as:low risk group(n=3),moderate risk group(n=17),high risk group(n=35)and very high risk group(n=34).Results The cfPWV in ISH patients is significantly higher than that of sys-diastolic hypertension group(ISH: 399.6?48.2 vs sys diastolic hypertension:374.3?39.7 cm/s,P0.05).The LF in ISH group are markedly higher than those in DH(ISH:4.35?1.07 log ms~2 vs 3.78?0.82 log ms~2,P 0.05).LF are markedly positive correlated with the degree of the total cardiovascular risk(rs=0.438,P

BACKGROUND: Many data demonstrate that the components of soybean can lower blood lipid,suppress the growth of cancer cells and exert weak estrogenic activities. However,little is known about the effect of isolated soybean protein on the lifespan of the organism.OBJECTIVE: To observe the effect of isolated soybean protein on the lifespan of drosophila melanogasters and investigate the mechanism of effective anti-aging and anti-oxidation action.DESIGN: A controlled trial based on drosophila melanogasters.SETTING: Department of nutrition and food hygiene in a university.MATERIALS: The experiment was conducted in the Department of Nutrition and Food Hygiene,Public Health College of Xinjiang Medical University from March to June 2002. A total of 400 drosophila melanogasters of American wild type with half for each gender were provided by the Department of Nutrition and Food Hygiene, Public Health College of Xinjiang Medical University.METHODS: The 400 drosophila melanogasters were divided into control group(normal culture) and three-dosage experiment groups(normal culture contained isolated soybean protein 0.2%,1.0% and 5.0%,respectively).From the second day on, the number of living and dead drosophila melanogasters was observed and counted until all died. Meanwhile, mean lifespan,half death time and maximal lifespan were calculated.MAIN OUTCOME MEASURFS: Mean lifespan, half death time and maximal lifespan of the drosophila melanogasters.RESULTS: Compared with that of control group, the lifespan of male and female drosophila melanogasters in experiment groups was prolonged by isolated soybean protein and responded in a dose-dependent manner,especially in high-dosage group. The mean lifespan, half death time and maximal lifespan of both female and male drosophila melanogasters were prolonged by 24.5% and20.7%,27.1% and22.0%,and 13.9% and 10.6%,respectively.CONCLUSION: Isolated soybean protein may have anti-aging and lifespan-prolonging effects on drosophila melanogasters.