4.Expression of TIMP-2 in peripheral blood lymphocytes in patients with chronic obstructive pulmonary diseases
zhen-ping, HU ; yi-cheng, ZHANG ; jun-gang, XIE
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To explore the expression of tissue inhibitor of metalloproteinases-2(TIMP-2) in peripheral blood lymphocytes in patients with chronic obstructive pulmonary diseases(COPD) and the association of smoking and TIMP-2 mRNA. Methods The expression of TIMP-2 mRNA in peripheral blood lymphocytes was measured by reverse transcription-polymerase chain reaction(RT-PCR) in 44 patients with COPD and 42 healthy smokers.The correlation analysis was then conducted between TIMP-2 mRNA expression and smoking index. Results The expression of TIMP-2 mRNA was 0.753?0.154 and 1.170?0.196,respectively,in patients with COPD and healthy smokers(P
6.Ilizarov technique used in treatment of bone and soft tissue defects of the tibia
Yijun REN ; Li YAH ; Rui HU ; Xincheng YI ; Wenjun CHENG
Chinese Journal of Orthopaedic Trauma 2017;19(3):213-218
Objective To report our clinical outcomes of treating tibial defects combined with soft tissue defects using Ilizarov technique.Methods From May 2010 to February 2015,52 patients with combined bone and soft tissue defects of the tibia were treated at our department.They were 41 males and 11 females,aged from 19 to 65 years (average,37.7 years).By Gustilo classification,49 cases were type ⅢB and 3 type ⅢC.The areas of soft tissue defect ranged from 7 cm ×3 cm to 28 cm × 15 cm,and the tibial defects ranged from 5 cm to 15 cm in length (average,12.6 cm).The schemes of Ilizarov technique depended on the location and size of the tibial defects.Open wound dressing combined with bone transport was adopted in 21 cases,limb shortening followed by bone lengthening with compression at the fracture ends in 12 cases,and tissue flap transplantation combined with bone transport or lengthening in 19 cases.Results The follow-up time of the 52 patients ranged from 13 to 61 months (average,27.1 months).The distance of bone transport or lengthening ranged from 5.0 cm to 13.6 cm (average,10.8 cm);the bone transport speed averaged 0.81 mm/day.The tibiae united in all the 52 patients;the time for external fixation ranged from 13 to 21 months (average,15.3 months);the external fixation index was 2.3 months/cm.According to the Paley functional criteria,23 cases were excellent,19 good,9 fair,and one poor,yielding an excellent to good rate of 80.7%.Conclusion According to the location and size of the bone and soft tissue defects of the tibia,the 3 schemes of Ilizarov technique can be rationally chosen to obtain fine clinical outcomes.
8.Establishment and optimization of real-time PCR with SYBR Green Ⅰ for quantification of survivin
Li-Hua HU ; Zheng-Jiang CHENG ; Yi-Rong LI ; Shao-Jun HUANG ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.
9.Serum Uric Acid and Prehypertension
Tao CHEN ; Wei LI ; Bo HU ; Yang WANG ; Xiao-Ru CHENG ; Xuan JIA ; Yi SUN ;
Chinese Journal of Hypertension 2006;0(08):-
Background Previous experimental and clinical studies have proved that elevated serum uric acid increased risk for developing hypertension.Whereas,there are a paucity of information on the relationship between serum uric acid and prehypertension.Objective The purpose of this research is to evaluate the association between the serum uric acid and prehypertension.Method A cohort of seven thousand eight hundred thirty-nine subjects without hypertension and other cardiovascular diseases were recruited from a cross sectional study in urban and rural place in 9 provinces during 2005-2006.Based on serum uric acid(324 ?mol/L for overall population,366 ?mol/L for male,285 ?mol/L for female),people were categorized into quartiles.The odds ratio for prehypertension was calculat ed with the lowest quartile as the reference.Results The prevalence of prehypertension increased with increasing uric acid in total population(P324 ?mol/L)to lowest quartile 1(
10.Creation and Characteristics of HepG2/mdr1 Cell Line
Ju ZHANG ; Jie CHENG ; Jing CHEN ; Juan YI ; Jing SUN ; Hu-Lai WEI ;
China Biotechnology 2006;0(05):-
Eukaryotic expression plasmid pCI-neo-mdr1 which contains human multidrug resistance gene 1(mdr1),was constructed and transferred into human hepatocarcinoma HepG2 cells by use of liposome. G418 was used to screen the cells successfully with mdr1 and the selected cells was named HepG2/mdr1 Morphological and biological properties of HepG2/mdr1 cells were observed. The results show that the constructed HepG2/mdr1 cell line was high efficient and stationary in the expression of mdr1. The work was valuable and desirable for the establishment of multidrug resistant cell models,and for the study of MDR in human hepatoma. Furthermore,the work also provided a perfect model for the research of relationship between insulin resistance and MDR in hepatocarcinoma cells.