1.Expression of THANK in human PBMC activated with different stimulators
Dong WU ; Feng SHEN ; Yonghua LOU ; Binghua JIAO ; Mengchao WU
Journal of Cellular and Molecular Immunology 2001;17(4):304-306
Aim To analyze THANK gene expression in peripheral blood mononuclear cells(PBMC) stimulated with different stimulators and to clone whole length human THANK gene. Methods PBMC were conventionally isolated and cultured in RPMI1640 containing 10% FCS. After stimulated with LPS,TNF α ,IL 2,IFN γ ,PHA or PMA,the THANK gene expression in PBMCs was analyzed by RT PCR and THANK cDNA was cloned. Result RT PCR detection showed that THNAK gene expressed in PBMCs after stimulated with interferon γ for 3 days, whereas THANK'expression could not be detected after stimulated with LPS,TNF α ,IL 2,IFN γ ,PHA or PMA respectively. Then THANK gene was cloned by cloning PCR product and sequenced. Conclusion Human THANK gene is cloned successfully, thus providing the possibility for further research of THANK'function.
2.Expression, denaturation, renaturation and purification of THANK protein
Dong WU ; Feng SHEN ; Yonghua LOU ; Min PENG ; Binghua JIAO ; Mengchao WU
Academic Journal of Second Military Medical University 2001;0(09):-
Objective: To prepare highly purified THANK protein. Methods: THANK was efficiently expressed in E.coli as inclusion bodies. After bacteria were lysed under ultrasonication, TE buffer,1% TritonX and 2 mol /L urea was used to efficiently extract inclusion bodies. After denaturation with 8 mol/L urea,THANK was partially purified by Sephacryl S-200 chromatography, and then subsequent refolding step was optimized for a maximal recovery of biological active protein. The renatured THANK was purified to homogeneity with Q Sephadex Fast Flow gel filtration and then desalted by Sephadex G-25 chromatography. Results: The purity of biologically active THANK was above 97% in SDS-PAGE densitometric studies. Conclusion: An effective method of denaturation, renaturation and purification of recombinant THANK is established.
3.The clone and expression of human THANK gene
Dong WU ; Feng SHEN ; Yonghua LOU ; Yuping ZHU ; Binghua JIAO ; Mengchao WU ;
Academic Journal of Second Military Medical University 2000;0(07):-
Objective: To clone THANK gene and express its extracelluar fragment. Methods: Using RNA isolated from HL 60 cell lines, THANK cDNA was amplified by RT PCR. The fragment was linked to pMD18 T vector and sequenced, and then the extracellular fragment of THANK was subcloned into pET vector and THANK protein expression was induced.Results: A 858 bp DNA fragment was amplified and the cDNA sequence was identical with the published sequence encoding THANK gene. Western blot showed that THANK protein with a relative molecular weight of 2.6?10 4 was expressed. Conclusion: Human THANK gene was cloned and expressed successfully, which provides a base of further research of THANK gene. [
4. Application of narrow-band imaging without magnifying in diagnosis of colorectal lesions by NICE classification
Journal of Shanghai Jiaotong University(Medical Science) 2019;39(2):176-181
Objective: To evaluate narrow-band imaging (NBI) without magnifying in the diagnosis of colorectal lesions by NBI International Colorectal Endoscopic Criteria (NICE classification), and analyze the safety and practicability of "do-not-resect" and "resect and discard" policies in clinical practice. Methods: The patients undergoing screening or surveillance colonoscopy, who were found colorectal lesions in the examination, from May to December in 2017 were enrolled. All the patients were examined by NBI without magnifying by any of the designated two physicians. NICE classification was used to diagnose colorectal lesions, and the diagnostic confidence of each lesion was recorded. The results of endoscopy were compared with those of pathology, and the accuracy rate and the confidence rate of diagnosis were calculated. The sensitivity, specificity, positive predictive value and negative predictive value of the diagnostic method for differentiating superficial tumors from non-tumors were also calculated. Finally, the feasibility, safety and cost savings of using "do-not-resect" and "resect and discard" policies in clinic were analyzed. Results: A total of 764 lesions were detected in the 636 enrolled patients. The overall accuracy of NICE classification was 84.95% and the diagnostic confidence rate was 81.68%. The sensitivity, specificity, positive predictive value and negative predictive value for differentiating tumors from non-tumors were 91.77%, 67.68%, 88.69%, and 74.86%, respectively. The diagnostic accuracy of diminutive colorectal lesions (≤5 mm) with high confidence was 94.98%, and the negative predictive value of diminutive rectosigmoid lesions (≤5 mm) with high confidence was 96.25%. They achieved the criteria of "resect and discard" and "do-not-resect" policies. If "do-not-resect" and "resect and discard" policies had been executed in clinical practice, ¥165 490 could have been saved and the omission diagnostic rates of "do-not-resect" and "resect and discard" policies would have been 3.75% and 0, respectively, in this study. Conclusion: It is feasible to use NBI without magnifying in differentiating tumors from non-tumors. The diminutive colorectal lesions and rectosigmoid lesions with high diagnostic confidence may achieve the criteria of "resect and discard" and "do-not-resect" policies, respectively.
5.Analysis of time trend of hepatocellular carcinoma mortality in Haimen city of Jiangsu province from 1993 to 2006
Jiao-Feng WANG ; Wen-Yao LIN ; Feng JIANG ; Wei MENG ; Fu-Min SHEN
Chinese Journal of Epidemiology 2010;31(7):727-732
Objective To predict the trend of hepatocellular carcinoma (HCC) mortality and investigate the features of its mortality including age, period, and birth cohort in males living in Haimen city of Jiangsu province, China. Methods Grey model (GM) was modeled using standardized mortality rate (SMR) of HCC from 1993 to 2006, and was applied to predicting SMR until 2012. Based on the mortality density (MD) for a four-year period, the goodness-of-fit of models and comparisons between models were evaluated so as to obtain the best one among these models including the effects of intercept, age-period-cohort (APC) , age-period (AP), age-cohort (AC),period-cohort(PC), and APC. Both APC full model and the best model were used to estimate effects of age, period, and cohort on HCC mortality. In addition, MD from 2005 to 2012 was predicted by the best model. Results Predictions based on GM (1,1 )showed that SMR was 48.578 per 100 000 population (relative error=-1.267% ) in 2007 year, which declined between 2008 and 2012. The lowest value was 45.578 per 100 000 people (in the 2012 year). The results of fitted models and comparisons between models showed that AP model was the best one (△G2=9.065,AIC=202.544). The curvatures of the effects of the three factors from APC model suggested that significances existed in changes of curvatures of 36.5-40.5 years old- (-0.368) and 64.5-68.5 years old-(-0.489) as well as in the change of 1956-1959 birth cohort (C21949.5. 1967.5=-0.492). The estimation of relative risks for AP model showed that the age effects were upward to 64.5-68.5 years old-, then downward; and that the period effects were found to be declined between 1993 and 2004. Predictions based on AP model suggested the decrease of HCC mortality. Conclusion The slightly decreasing trend of HCC mortality for males might be explained by age, period and a minor birth cohort effects in Haimen of China.
7.Evolution of interventional cardiology in China.
Chinese Medical Journal 2009;122(19):2209-2219
8.Genetic polymorphism for 124 Individual Identiifcation SNPs from Chinese Han using Ion PGM? platform
He SONG ; Yishu ZHOU ; Feng LIU ; Hongying SHEN ; Jiao YU ; Jinling ZHAO ; Bin ZHAO ; Fei GUO ; Xianhua JIANG
Chinese Journal of Forensic Medicine 2016;31(4):345-350
ObjectiveTo investigate the polymorphisms of 124 individual identiifcation SNPs in Chinese Han using the Ion Personal Genome Machine?(PGMTM).Method Samples from 130 unrelated Chinese Han individuals and two families (8 genealogical individuals) were ampliifed using Ion AmpliseqTM Library kit and sequenced on Ion Torrent PGM? platform.Results 14 148 SNPs were detected.A total of 99.992 9% SNPs were correctly called by the HID SNP Genotyper v4.3 plugin, while 0.007 1% wrongly reported and 62 NN calls needed manual correction. The MP ranged from 0.348 0 (rs2831700) to 0.817 3 (rs740910) with the value of 6.898 4 × 10-34 for CMP. The DP ranged from 0.182 7 (rs740910) to 0.652 0 (rs1355366) with the value of 0.999 999 999 999 999 999 999 999 999 999 999 310 2 for CDP, which was larger than that of 22 STR loci. The PE ranged from 0.007 3 (rs1024116) to 0.278 1 (rs1058083) with the value of 0.999 999 616 7 for CPE, which was smaller than that of 22 STR loci. A total of 8 Y-SNP haplo-types were observed from 72 unrelated male samples. No mutation was observed from pedigrees.Conclusion The 124 IISNPs were high polymorphic in Chinese Han and they were ideal markers for human identiifcation. The PGMTM platform has a potential role in forensic science.
10.Optimization of atrioventricular delay by surface electrocardiography during dual chamber pacing.
Li-qun WU ; Gang GU ; Min CAO ; Yong-chu SHEN ; Kan SU ; Wei-feng SHEN
Chinese Medical Journal 2006;119(6):454-457
BACKGROUNDNominal atrioventricular (AV) interval in dual chamber pacemaker (DDD) is not the best AV delay in the majority of patients with atrioventricular block. To find a simple method for optimizing AV delay adjustment, we assessed surface electrocardiography (ECG) for optimizing AV delay during dual chamber pacing.
METHODSDDD pacemakers were implanted in 46 patients with complete, or almost complete, AV block. Optimal AV delay was achieved by programming an additional delay of 100 ms, to the width of intrinsic P wave or to the interval between pacing spike to the end of P wave on surface ECG. Left ventricular (LV) end diastolic and end systolic volumes, ejection fraction and diastolic parameters were measured by Doppler echocardiography during both nominal and optimal AV delay pacing.
RESULTSCompared to nominal AV delay setting, LV end diastolic volume increased [to (53.2 +/- 11.3) ml from (50.2 +/- 10.2) ml, P < 0.05], end systolic volume decreased [to (26.1 +/- 9.0) ml from (27.9 +/- 8.2) ml, P < 0.05] during adjusted AV delay pacing, resulting in an increase in LV ejection fraction [to (68.2 +/- 5.3)% from (64.5 +/- 4.3)%, P < 0.05]. LV diastolic filling and isovolumic relaxation time were not significantly changed.
CONCLUSIONOptimization of AV delay by surface ECG is a simple method to improve LV systolic function during dual chamber pacing.
Adult ; Aged ; Aged, 80 and over ; Atrioventricular Node ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Electrocardiography ; methods ; Female ; Heart Block ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Time Factors