1.DETERMINATION OF ELEVEN ELEMENTS IN BLOOD OF THE NORMAL AND PATIENTS IN NANJING
Acta Nutrimenta Sinica 1956;0(01):-
The elements in blood or serum, including calcium, magnesium, copper, zinc, chromium, manganese, nickel, selenium, lead and cadmium were determined in the normal and patients in Nanjing by atomic absorption spectrophotometry. The method used was proved to be reliable being checked by the reference bovine serum and the data obtained in this paper was coincident with those reported by the other authors previously. The relationship between the concentrations of elements and the ages as well as the sexes in the normal was discussed briefly. The concentrations of several of the elements of the patients were significantly different from those of the normal in the same ages.
2.Antitumor efficacy of specific cytotoxic T lymphocytes activiated by dendritic cells pulsed with K-ras antigen to pancreatic cancer ex vivo
Li XU ; Yanqiu FANG ; Guang TAN ; Zhongyu WANG ; Yan TAN
Chinese Journal of Immunology 2009;25(12):1075-1079,1091
Objective:To investigate the antitumor efficiency of the special cytotoxic T lymphocytes(CTLs) activated by dendritic cells(DCs) pulsed with K-ras (12-Val) antigen.Methods:DCs was generated from PBMC in the presence of granuloceyte/macrophage-colony stimulating factor(GM-CSF),interleukin-4(IL-4)in vitro.DCs were differently sensitized with K-ras mutant pancreatic cancer cell line,K-ras(12-Val) mutant peptide,K-ras(12-Val) mutant peptide with the surface of cationic nanoparticle.Cell surface markers on DCs was measured by flow cytometry.The activation of CTL induced by DCs was detected by ~3H- thymidine incorporation test.The killing effects of CTL to pancreatic cancer was detected by ~(125)I-UdR release test. Production of IL-12 and IFN-γ by DCs and PBMC was detected by ELISA.Results:Compared with DCs pulsed with K-ras(12-Val) mutant peptide and K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle,DCs pulsed with whole tumor antigen could better induce CTLs killing activity(P<0.05).The DCs with K-ras(12-Val) mutant peptide and K-ras mutant peptide with the surface of cationic nanoparticle could produce specific CTL killing activity aganist pancreatic cancer cell line Patu8988(K-ras+)(P<0.05),but not SW1990(K-ras-)(P>0.05). K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle at lower concentrations can be effectively presenting on the surface of DCs than only K-ras (12-Val) mutant peptide.Conclusion:K-ras (12-Val) mutant peptide with cationic carrier can be effectively presenting and expression of DCs and induce CTL specific killing activity aganist pancreatic cancer cell lines with K-ras (12-Val) mutant peptide.
3.Genetic Analysis of VP_1 Sequences of Enterovirus 71 Isolated from Patients of Hand,Foot and Mouth Disease in Beijing,2008
Fang HUANG ; Weihong LI ; Xiaojuan TAN
Chinese Journal of Vaccines and Immunization 2008;0(06):-
Objective To get genetic information of VP 1 coding region of HEV 71 in Beijing in 2008.Methods Enteroviruses were isolated from samples of throat swabs collected from 33 HFMD patients within 3 days after onset by rhabdomyosarcoma(RD) cells and identified by RT-PCR method with specific primers to human enteroviruses,then VP 1 coding region was amplified and sequenced by Sanger dideoxy.Bioedit 7.0.5 and MEGA3.1 were used for the nucleotide sequence alignment and phylogenetic analysis.Results 16 virus strains were isolated from 33 samples,of which 14 strains were identified as HEV 71 and 2 were CVA 16,and 1 was co-infected with CVA 16 and HEV 71.Sequence analysis of VP 1 nucleotide sequences of 10 HEV 71 isolates showed that homologous analysis of nucleotide identity amino acid 95.5%~100% and 98.9%~100% respectively;representative strains of Fuyang in 2008,nucleotide identity was 95.4%~99.1%;with representative strain of C 4 nucleotide identity was over 92%.Phylogenetic analysis of HEV 71 strains for the nucleotide sequence of VP 1 coding region clarified that the HEV 71 isolates in Beijing belonged to C 4a cluster of C 4 sub-genotype and 10 strains formed four relatively separated clusters.Conclusions The HEV 71 viruses isolated from children of HFMD in Beijing belonged to C 4 sub-genotype,and C4a cluster which was the predominant in China since 2004.According to phylogenetic analysis,HEV 71 which belonged to more than 4 different clusters were circulating in Beijing in 2008.More virological suggestion for disease control and prevention,and information of HEV 71 molecular epidemiology need to be collected urgently due to the successive large epidemic of HEV 71 in China.
4.Effect of Gypenosides on Lipid Peroxidation in Rats with Type 2 Diabetes Mellitus Complicated by Nonalcoholic Fatty Liver Disease
Qin HE ; Fang LI ; Huabing TAN
Herald of Medicine 2014;(12):1549-1553
Objective To obserVe the effect of gyPenoside on liPid Peroxidation and hePatic lesion in rats with tyPe 2 diabetes mellitus and nonalcohol fatty liVer disease. Methods Totally,65 SPF male SD rats were randomly diVided into blank control grouP (grouP N),NAFLD model grouP (grouP NM),and NAFLD with T2DM model grouP. The NAFLD with T2DM model grouP was further diVided into three subgrouPs:JH grouP,Perfused with 1 g·kg-1 ·d-1 GPS;JL grouP,Perfused with 0. 5 g·kg-1 ·d-1 GPS;model control grouP,Perfused with the same Volume of water. Blood sugar,triglycerides ( TG) ,total cholesterol ( TC) ,alanine aminotransferase ( ALT) ,asPart aminotransferase ( AST) ,adePonectin ( ADP) in the Plasma were measured. TG, malondialdehyde (MDA),and suPeroxide dismutase (SOD) in the liVer tissue were also tested. Results ADP leVel was (7. 46±1. 12),(3. 58±0. 98),(4. 89±1. 02),(4. 79±1. 01) and (4. 13±0. 89) ng·mL-1 in N,M,NM,JH and JL grouPs, resPectiVely. The ADP leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). MDA leVel was (2. 98±0. 09),(4. 22±0. 11),(3. 66±0. 10),(3. 72±0. 11),(3. 99±0. 13) nmol·mL_1 in N,M,NM,JH and JL grouPs,resPectiVely. The MDA leVel was significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). SOD leVel was (240. 8±17. 4), (149. 9±20. 6),(181. 6±19. 4),(209. 8±19. 2),(189. 4±18. 9) U·mL_1 in N,M,NM,JH,and JL grouPs,resPectiVely. SOD leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). TG leVel was (28. 98±1. 68),(214. 46±5. 44),(198. 46±6. 98),(142. 87±6. 64) and (164. 92±7. 56) mg·g-1 in N,M,NM,JH and JL grouPs,resPectiVely. TG leVel was significantly lower in grouP JH and JL than in grouP M ( P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). ALT and AST were significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). Conclusion The liPid Peroxidation in the liVer of rats with T2DM comPlicated with NAFLD can be reduced by gyPenoside,and hePatic lesion may be alleViated through inhibition of liPid Peroxidation.
5.Treatment of patients with hypoparathyroidism accompanied by elevated serum muscle enzymes
Qiang TONG ; Hong LI ; Xiaozheng FANG ; Juan TAN
Chinese Journal of Endocrinology and Metabolism 2013;29(9):798-800
The clinical data of patients with hypoparathyroidism accompanied by elevated and normal muscle enzymes were analyzed retrospectively.The results showed that there were no differences in age,gender,pathogenesis,etiology,and parathormone level between groups with elevated muscle enzyme and normal muscle enzyme.Serum calcium concentration in the group with normal muscle enzyme was within normal range.Serum calcium concentration in the group with elevated muscle enzyme was significantly lower than that in the group with normal muscle enzyme (P<0.01).As the serum calcium concentration rose,the elevated enzyme level gradually returned to normal range,suggesting that the decrease of serum calcium concentration resulted in the elevated muscle enzyme levels in patients with hypoparathyroidism.
6.Hepatitis B virus X gene transforms human hepatocyte QSG7701 in vitro
Tao LI ; Guozhen LIU ; Deming TAN ; Chuanxiang WU ; Fang ZHENG
Chinese Journal of Infectious Diseases 2009;27(7):398-402
Objective To observe the effects of hepatitis B virus(HBV)X gene muhifunctional protein(HBx)on the biological characteristics of QSG7701 and the transformational effects on QSG7701 cell.Methods QSG7701 ceils were stably transformed by recombinant plasmid pCMV/X and eukaryotic expressed plasmid pRc/CMV2 by liposome-based assay,respectively.Non-transfeeted QSG7701 cells were employed as controls.The expressions of HBx,c-Myc and Bel-2 proteins in QSG7701 cells were detected by Western blot.MTT colorimetric analysis,flow cytometry and soft agar clone-forming assay were performed tO detect the biolo~dcal activity of cells.Results HBx Drotein was highly expressed in pCMV X/QSG7701 cells.The expression level of c-Myc protein in the pCMV X/QSG7701 cells was much higher than those in the other tWO groups of cells.The expression of Bcl2 protein was detected in the three groups of cells,but the expression levels were similar.Percentage of S stage cells in pCMV X/QSG7701 ceils was significantly higher than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells E(28.80±2.32)%,(15.5±2.64)%and(21.5±3.66)%,LSD 0.05=3.95%,LSD 0.01=5.47%,P<0.01].While percentage of G1 stage cells in pCMV X/QSG7701 cells was significantly lower than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells[(62.30±3.85)%,(78.70±4.12)%and(78.10q±4.45)%,LSD 0.05=5.63%,LSD 0.01-7.79 %,P<0.01].The apoptosis rate of pCMV X/QSG7701 cells was much higher than those in pRcCMV2/QSG770t and non-transfected QSG7701 cells[(14.90±1.01)%,(8.91±0.48)%and (4.03±0.47)%,LSD 0.05=O.94%,LSD 0.01=1.31%,P<0.01].The population doubling time of pCMV X/QSG7701 cells was shorter than those in pRcCMV2/QSG and non-trarmfected cells(14 h,29 h,38 h,respectively).The cloning ratio in soft agar of pCMV X/QSG7701 cells WaS significantly higher than those of pRcCMV2/QSG and non-transfected QSG7701 cells[(19.83±1.96)%,(1.76±0.03)%and (1.33±0.18)%,LSD 0.05=1.53%,LSD 0.01=2.11%,P<0.01].Conclusion HBx may transform human non-tumor hepatoeyte QSG7701,which makes the cell growth malignizeA.
7.Adenovirns construction with miceCD40- IgG2aFc-IRS-GFP fusion gene and in vitro testing
Chuanguang LI ; Yifeng FANG ; Yingxia TAN ; Zhijian SHEN ; Qiyu ZHANG
Chinese Journal of Hepatobiliary Surgery 2010;16(3):208-211
Objective To construct the adenovirus vector containing mice CD40- IgG2aFc-IRS-GFP fusion gene and detect the expression of the fusion protein in transfeete 293cells.Methods The fragments of CD40, IgG2aFc were obtained and inserted into plasmid PDC316-IgG2aFc-GFP.After being verified by sequencing, Ad5-PDC316-CD40-IgG2aFc-GFP was contransfored with the bckbone vector into 293 cells.The virus titer was detected after replicating and purifing and the expression of the fusion protein was analyzed.Results A virus and plasmid were constructed successfully.The vitro infection showed that the virus can infect cells of which the fusion protein was confirmed by fluo-rescence.The expression of the fusion protein increased with the increased time and virus concentra-tion.The protein expression stopped increasing after the virus concentration came to a certain level.Conclusion CD40, IgG2aFc fragments are correctly ligated with plasmid PDC316-IgG2aFc-GFP, and the fusion protein can be expressed in 293cells.This might lay a foundation for further studies of the expresstion of virus, immune tolerance and mechanism of liver transplantation model in rats.
9.A preliminary study on the establishment of rat model with acute hyperuricemia
Li XU ; Le SHI ; Fang ZHAO ; Qiuwei TAN ; Jinliang YAO
Chinese Pharmacological Bulletin 2003;0(07):-
Aim To observe the fluctuation of uric acid in serum after replicating rat hyperuricemia model with hypoxanthine, uricase inhibitor singly or combined. Discuss the method of acute hyperuricemia replication on rats. Method Hypoxanthine was administered via stomach, uricase inhibitor was injected intraperitoneally, then inspect the uric acid in serum at different periods of time. Results When we combined hypoxanthine with uricase inhibitor, the uric acid in serum of different dose groups was higher than control group after 3 hours(P
10.Study on hNIS mediated transferring gene99mTcO-4 imaging and mI treatment in xenografted ovariancancer
Jingqiu CUI ; Peihua FANG ; Ning LI ; Ping FENG ; Jian TAN
Chinese Journal of Endocrinology and Metabolism 2009;25(1):83-86
Objective To establish radioiodine therapy in nonthyroid tumor and to investigate 131Ⅰ treatment effect on xenografted ovarian cancer. Methods Based on previous test, xenografted ovarian cancer nude model were established in nude mice. The effects of radioactive isotope 99m TcO-4 imaging and radioiodine 131Ⅰ treatment on xenografted ovarian cancer in vivo were investigated. Results After transferring human sodium/iodide symporter (hNIS) gene, the xenografted ovarian cancer in nude mice was imaged by isotope 99m TcO-4 Moreover,131Ⅰ exerted inhibitory effect on the proliferative activity. Conclusion After the transfection of hNIS gene, 131Ⅰ has inhibitory effect on proliferative activity of xenografted ovarian cancer.