4.Comparision of the Efficiency of Different Promoters Driving the Expression of CD19-specife Chimeric Antigen Receptor (CAR)-redirected T Lymphocytes Mediated by Lentiviral Vector
Journal of Medical Research 2017;46(6):92-98
Objective Three kinds of different internal promoters were inserted into the lentiviral vector to drive the expression of CD19-specifc chimeric antigen receptor (CAR)-redirected T lymphocytes with the green fluorescent protein(GFP) and the GFP expression efficiency of transduced cells driven by lentivirus were compared.Methods Three GFP reporter lentivirus vectors carrying different promoters and anti-CD19-CAR,including CMV,EF1α,and CMV-EF1α were selected.Three different vectors' names abbreviate to pHAGE-CMV-EFlα-EGFP-CD19,pHAGE-CMV-EGFP-CD19,pHAGE-EF1α-EGFP-CD19.Human embroic kideny 293T cells were cotransfected with the three plasmids by calcium phosphate DNA precipation.And the expression of GFP was observed under fluorescent microscope after transfecting 293T cells,and virus supernatant was collected after 72 h and centrifuged.Nucleic acid copy number in RNA and the expression of EGFP and CD3 zeta were identified through fluorescence quantitative PCR,flow cytometry and western blotting,respectively.The titers of the lentiviral vectors were determinde by scoring GFP expression following swerial dilutions of the viral supernatant on 293T cells.T lymphocytes was infected with lentivims produced from these reporter vectors.Then,fluorescence microscope,wesyern blotting were used to detect the GFP expression strength.Results Results of fluorescence microscope maintained that different internal promotors driving the expressing effect is different.293T as targeted cell,the transfected 293T cells were found containing strong expression of GFP.The amount of the 293T cells expressing GFP driven by CMV-EF1α promoter was the largest,while the EF1α promoter was the smallest.Flow cytometry results show that the rate of transduction is 72.4%,20.6% and 14.5%,respectively.T lymphocytes as targeted cell,Western blot showed that CMV-EF1α was the largest among all promoters,while the CMV was the smallest.Conclusion In the study of gene expression mediated by lenuvirus,proper cell lines and promoters should be selected to obtain high efficiency.
5.Investigation on Key Aspects of the Bilingual Teaching in Microbiology
Microbiology 1992;0(02):-
Fundamental specialized courses are important due to their connection between fundamental knowledge and major knowledge,which are regarded as the difficult courses for undergraduates to learn.The useful approaches and experience of bilingual teaching in microbiology,one of the fundamental specialized courses for the students majoring in Bioengineering,are presented here.The key aspects such as choosing textbooks,teaching manners and teaching contents are also discussed.This paper also proposes how to improve the quality of bilingual teaching during the fundamental specialized courses.
6.Sensory reeducation for restoring the sensation of palm after the median nerve injuries
Xiufu LAN ; Yueqiu LIN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(06):-
Objective To observe the effects of early rehabilitation training on sensory restoration after the median nerve injuries. Methods Seventy patients with median nerve injuries were randomly divided into two groups: the rehabilitation group ( n =32), in which the patients received sensory reeducation two weeks after surgical treatment; and the control group ( n =38),in which the patients only received surgical treatment. After 6 8 weeks of treatment, reassessment were performed and the results were compared between the two groups. Results The excellent and effective rates of functional recovery in sensation in the rehabilitation group were 88% and 97%, respectively. A comparison showed that there was significant difference between the two groups. Conclusion Sensory reeducation could accelerate the restoration of sensation in patients with median nerve injury.
7. Optimization of urinary sample preparation process for real-time PCR detection of BK virus
Academic Journal of Second Military Medical University 2010;31(4):438-441
Objective: To develop an effective preparation method to improve the sensitivity and accuracy of real-time PCR in detection of BK virus's (BKV's) load in urine samples. Methods: A total of 24 samples documented as positive probes in primary detection were enrolled in this study. The candidate samples were prepared by 4 different approaches: unprocessed urine, BKV's DNA extracted from urine, 1:10 diluted urine, and 1:100 diluted urine; and then they were subjected to real-time PCR examination to obtain the viral load. The data obtained were analyzed by SPSS 11.0. Results: The four different preparation processes for urinary specimens had significant impact on detection results of real-time PCR. Three samples were negative in the unprocessed urine group and 66.7% of its samples had the lowest viral loads compared with the other three groups. Two samples in the 1:100 diluted urine group were negative and 79.2% of its samples had the highest viral loads, but its median load was similar to that of the 1:10 group. Viral gene was detected in all samples in the DNA extraction group and 1:10 diluted urine group, but the loss of the target gene was more severe in the DNA extraction group. Conclusion: The 1:10 diluted urine is better for real-time PCR detection of BKV's load, as it lose less viral gene and is more efficient, easy to perform and economical.
8.TCM and western medicine research progress of diabetic peripheral neuropathy
International Journal of Traditional Chinese Medicine 2010;32(6):558-560
Diabetic peripheral neuropathy (DPN) is a common complication of diabetic mellitus. The treatment is so difficult that doctors have not found reliable methods to cure this disease. The research progress both in TCM and western medicine of this disease were reviewed in terms of aspects of etiology, pathogenesis and differential treatments.
9.Application of the tissue chip in experimental teaching of morphology
Yonghong LAN ; Haiyan NIU ; Shizhen LIN
Chinese Journal of Medical Education Research 2011;10(12):1523-1524
The tissue chip is a special tissue section,which has several tens to thousands mini tissues of regular arrangement on the one glass slide.According to the deficiencies in experimental teaching section of morphology at present,the author introduced the characteristics and values of the tissue chip and explored its application prospect in experimental teaching of morphology.
10.Comprehensive evaluation on the curative effect of TCM treating type 2 diabetes with three type dialectical therapy by Fuzzy Mathematics
International Journal of Traditional Chinese Medicine 2011;33(11):972-975
ObjectiveTo comprehensively evaluate the curative effect ofTCM reating type 2 diabetes with three type dialectical therapy by Fuzzy Mathematics.Methods261 cases of 2 type diabetes were divided into three groups by syndrome differentiation,as follows:type of heat with yin deficiency,type of deficiency of both qi and yin,and type of deficiency of both yin and yang,and received corresponding therapies.The course of treatment was 24 weeks,in the first 12 weeks (period 1),on the basis of western medicine treatment,patients of the three groups were respectively treated by qingrungranules,ziyigranule and shuangtiaogranules.In the next 12 weeks (period 2),all patients were only treated by western medicine.The value of FBG,P2BG,HbAlc,TC,TG,BMI of the two periods were comprehensively evaluated by the methods of fuzzy mathematics.ResultsAt the end of period 1,the total effective rate of the type of heat with yin deficiency was 91.38%,the type of deficiency of both qi and yin was 87.16%,and the type of deficiency of both yin and yang was 83.34%; while at the end of period 2,the total effective rate of the three types was 59.18%,48.65%a nd32.65% respectively.ConclusionThe effective rate was obviously improved in period 1; while in period 2,the effective rate was decreased.