2.Cloning and expression of human glucagon-like peptide-1 cDNA
Zhi-Zhen ZHANG ; Ji-Fang MAO ; Hong DOU ; Sheng-Sheng YANG
Academic Journal of Second Military Medical University 2001;22(4):316-318
Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion: Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.
3.Cloning and expression of human glucagon-like peptide-1 cDNA
Zhi-Zhen ZHANG ; Ji-Fang MAO ; Hong DOU ; Sheng-Sheng YANG
Academic Journal of Second Military Medical University 2001;22(4):316-318
Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion: Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.
4.Therapy for hepatic encephalopathy.
Chinese Journal of Hepatology 2004;12(5):306-307
5.A comparison of four methods for extraction of human fecal DNA by using real time PCR
Zhong-Wen WU ; Ying HAN ; Hai-Feng LU ; Lan-Juan LI ; Ji-Fang SHENG ; Jian ZUO ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P
6.THE ROLE OF APP17 ON MODULATION OF IRS-1 AND IGF-1R EXPRESSIONS IN HIPPOCAMPAL NEURONS OF DIABETIC MICE
Pengwen WANG ; Shuli SHENG ; Fang YANG ; Zhijuan JI ; Zhigao JIN ; Zhil TAO
Acta Anatomica Sinica 1957;0(04):-
Objective To investigate the effect of a peptide,APP17,on regulating the expression of insulin receptor substrate\|1(IRS\|1) and insulin\|like growth factor (IGF\|1R) in neurons of the hippocampus from diabetic mouse. Methods Diabetic mouse models were established by injection of streptozotion.In experimental group,these models were injected with APP17 peptide subcutaneously and their brain sections were taken after 4 weeks of survival. The immunohistochemical stainning of these sections were then performed with IRS\|1 and IGF\|1R antibody.With regard to control groups,the mouse models were only injected saline and gone through the same procedure of immunohistochemistry together with normal mice. Results IRS\|1 and IGF\|1R positive neurons were widely distributed in the hippocampus of the diabetic mice,and the cytoplasm was darkly stained.In the contrast,positive cells in the hippocampus were lightly stained in those normal mice and the APP17 peptide\|treated diabetic mice. Conclusion The expression of IRS\|1 and IGF\|1R could increase in the hippocampus of dabetic mice.The APP17 can regulate the distribution of IRS\|1 and IGF\|1R in the brain of diabetic mice and return them to normal situation.
7.THE EFFECT OF APP17 ON MODULATION OF Bcl-2,Bax,CREB,AKt,AIF EXPRESSIONS IN HIPPOCAMPAL NEURONS OF D-GALACTOSE INDUCED BRAIN AGING OF THE MICE
Pengwen WANG ; Zhiwei ZHAO ; Lina SONG ; Fang YANG ; Zhijuan JI ; Shul SHENG
Acta Anatomica Sinica 1955;0(03):-
Objective To investigate the effect of the peptide APP17 on regulating the expression of Bcl\|2,Bax,cAMP response element binding Protein(CREB),Ser\|Thr kinase B/protein kinase B(Akt/PKB),apoptosis inducing factor(AIF) in neurons of the hippocampus from the D\|gal mouse. Methods D\|gal mouse models were established by injection of D\|gal.In experimental group,these models were injected with APP17 petide subcutaneously and their brain sections were taken after 3 months of survival.The immunohistochemical staining of these sections was then performed with Bcl\|2,Bax,CREB,Akt,AIF antibody. Results Bax,AIF positive neurons were widely distributed in the hippocampus of the D\|gal mice,and the cytoplasm was darkly stained.In contrast,positive cells in the hippocampus were poorly stained in those normal mice and the APP17 peptide\|treated D\|gal mice.But Bcl\|2,CREB,AKt positive neurons were widely distributed in the hippocampus of those normal mice and the APP17 peptide\|treated D\|gal mice,and the cytoplasm was darkly stained.In contrast,positive cells in the hippocampaus were poorly stained in the D\|gal mice.Conclusion\ The expression of Bax and AIF could be increased in the hippocampus of D\|gal mice.But the expression of Bcl\|2,CREB,AKt decreased in the hippocampus of D\|gal mice.The APP17 can regulate the distribution of Bcl\|2,Bax,CREB,Akt,AIF in the brain of D\|gal mice and return them to normal situation.\;[
8.Research and realization of signal processing algorithms based on FPGA in digital ophthalmic ultrasonography imaging.
Simin FANG ; Sheng ZHOU ; Xiaochun WANG ; Qingsheng YE ; Ling TIAN ; Jianjun JI ; Yanqun WANG
Chinese Journal of Medical Instrumentation 2015;39(1):1-4
OBJECTIVETo design and improve signal processing algorithms of ophthalmic ultrasonography based on FPGA.
METHODSAchieved three signal processing modules: full parallel distributed dynamic filter, digital quadrature demodulation, logarithmic compression, using Verilog HDL hardware language in Quartus II.
RESULTSCompared to the original system, the hardware cost is reduced, the whole image shows clearer and more information of the deep eyeball contained in the image, the depth of detection increases from 5 cm to 6 cm.
CONCLUSIONThe new algorithms meet the design requirements and achieve the system's optimization that they can effectively improve the image quality of existing equipment.
Algorithms ; Data Compression ; Diagnostic Imaging ; Ophthalmology ; Signal Processing, Computer-Assisted ; Ultrasonography
9.Research progress of microtubule inhibitor-induced tumor cell mitotic catastrophe
Xiang ZOU ; Chao WANG ; Zhongyuan QU ; Yueni FANG ; Jiejing SHENG ; Yubin JI
Chinese Pharmacological Bulletin 2015;(12):1637-1640
Microtubule inhibitor has been a hot area of anticancer drugs research.Microtubule inhibitor exert an anti-tumor effect by promoting or inhibiting the microtubule aggregation to break the dynamic balance of microtubule,hindering the spindle forma-tion of tumor cells,and then blocking the process of cell divi-sion.Mitotic catastrophe is a cell death phenomenon that is caused by abnormal cell division and damage of spindle structure in cell mitosis phase.In recent years more and more attention has been paid to mitotic catastrophe cell death because it has
been confirmed clinically that microtubule inhibitors can induce mitotic catastrophe death of tumor cells.This paper reviews the latest research progress of microtubule inhibitors,and discusses the molecular mechanisms of mitotic catastrophe cell death tumor cells induced by microtubule inhibitors.
10.The analyse of effectiveness in HBeAg-positive chronic viral hepatitis B treated by adefovir dipivoxil combined with bicyclol.
Dai-Rong XIANG ; Ji-Fang SHENG
Chinese Journal of Experimental and Clinical Virology 2009;23(4):299-301
OBJECTIVETo analyze the efficacy of adefovir dipivoxil combined with bicyclol in treatment of HBeAg-positive chronic viral hepatitis B (CHB).
METHODSA total of 91 patients with CHB were randomized into experimental group and control group to be treated. The patients in experimental group (46 samples) received adefovir dipivoxil orally 10 mg daily and bicyclol orally 150 mg daily for 48 weeks and those in control group (45 samples) received adefovir dipivoxil orally 10 mg daily alone for 48 weeks. The serum aminotransferace (ALT/ AST), HBV-DNA, HBeAg/antiHBe were observed before and after treatment.
RESULTSCompared with pretreatment, the serum aminotransferace were all decreased obviously in two groups, the experimental group is better (P < 0.05). HBVDNA negative conversion rate was significantly higher in experimental group than in the control group (47.8% vs. 31.1%, P < 0.05). There were nostatistically differrence between the two groups in the portion of HBeAg loss rate and HBeAg seroconversion rate. There were no obvious adverse reaction in the study.
CONCLUSIONAdefovir dipivoxil combined with bicyclol is efective in the treatment of chronic hepatitis B.
Adenine ; administration & dosage ; analogs & derivatives ; Adolescent ; Adult ; Aged ; Biphenyl Compounds ; administration & dosage ; Drug Therapy, Combination ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Male ; Middle Aged ; Organophosphonates ; administration & dosage ; Treatment Outcome ; Young Adult