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2000 01 [Abstract][OnlineView][Download 60k] - Jin Ningyi, Liu Yi, Guo Zhiru et al (Institute of Changchun University of Agriculture and Animal Sciences, Changchun 130062)
Objective To express IBDV VP2/VP243 gene in recombinant FPV and study on the protection and immunogenicity of expressed product. Methods Two recombinant plasmids, pUTALacVP2 and pUTAlacVP0,were constructed by inserting VP243 and VP2 genes of IBDV double strand genome fragment A to the downstream of hybrid promoter of cowlpox virus inclusion body type A(ATI) and vaccinia virus early promoter p7. 5 respectively. CEF cells were transfected with the 2 plasmids, and 2 kinds of recombinant virus- es, vUTALacVP2 and vUTAlacVP0, were screened out by X-gal staining. Then the 2 kinds of viruses were used for immunizing SPF chickens aged 1 week, using routine vaccine as control. Results ELISA, SDS-PAGE and Western blot demonstrated that the expressed products could reacted with specific PcAbs to IBDV and induce protective antibodies in chickens. Before challenge with vvIBDV, the antibody level in control group were signif- icantly higher than that in experimental group. However, the level in experimental group was increased rapidly 5d after challenge. Conclusion Two kinds of rFPV, vUTALac VP2 and VP0, were highly expressed in CEF cells. However, the protective rates of the recombinant viruses were lower than that of routine vaccine.
2000 01 [Abstract][OnlineView][Download 293k] - Zhu Junming, Quan Jiawn, Chen Wei (Wuhan Institate of Biological Products, Wuhan 430060)
Objective To study on the effect of antisense c-myc gene plasmid on the proliferation and apoptosis of rat glioma cell C6. Methods The plasmid pCEP4ASCMR expressing rat antisense c-myc gene in eukaryocyte was transfected to rat glioma C6 cell strain using Iiposome FuGENE6 as media, and the results were analyzed by MTT method, cell colony-forming test in soft agar medium and acridine orange staining. Results Antisense c-myc gene plasmid DNA could inhibit the proliferation and induce the apoptosis of glioma cells, and liposome FuGENE6 could promote the transfection of the plasmid DNA. Conclusion Antisense c-myc gene plasmid can inhibit the excessive expression of c-myc gene and is helpful to the treatment of glioma.
2000 01 [Abstract][OnlineView][Download 301k] - Li Yuexi , Wang Xingtai, Tao Kaihua et al ( East- China Research Institute for Medical Biotechniques , Nanjing 210002)
Objective To obtain engineering E. coli for the high expression of human cytomegavirus gp52 gene. Methods The human cytomegalovirus (HCMV) gp52 gene fragment containing 2 dominant antigen dormains confirmed by computer analysis was cloned by PCR and inserted into plasmid vector pET28(a),then the recombinant plasmid was transformed to E. coli BL21 and purified by Sephacry1 S-200 and Ni-NTA Sepharose chromatographies. Results About 28% of ed soluble somatic proteins were expressed, the molecular weight of the expressed gp52 protein was abaut 21 000. After being purified, the purity of it reached 95%. SDS-PAGE profile showed a single protein band,and ELISA showed good antigenicity and specificity of the expressed protein. Conclusion The study is of great signficance in examination of HCMV, especially in pregnant women and blood donors.
2000 01 [Abstract][OnlineView][Download 351k] - Zhou Fuyuan, Sui Lili, Pan Wensheng et al ( Departmens of Infectious Diseases, Nanfang Hospital, Cuang zhou 510515)
Objective To establish an effective, rapid and simple system for the expression and purifi- cation of hepatitis B core antigen. Methods HBcAg was expressed and purified in prokaryocytes by pQE30 system. Results 26. 2% of total somatic protein was expressed, and about 5 mg of HBcAg was obtained from 500 ml of bacterial fluid. After being purified, the purity of HBcAg reached 91. 0%. Western blotting showed combining acitivity of the expressed HBcAg with polyclonal anti-HBc. Conclusion An effection, rapid and simple system for the expression and purification of hepatitis B core antigen was established.
2000 01 [Abstract][OnlineView][Download 278k] - Liu Zhu, Zhou Hua, Yao Weimin et al (Changchun Institute of Biological Products, Changchun 130062)
Objective To prepare -2/Pseudomonas exotoxin A (PEA) chimeric protein used for the treatment of related diseases. Methods After IL-2 and PEA genes were amplified by PCR, the 3' terminal of IL- 2 gene and 5' terminal of PEA gene were linked, then cloned into vector pBV220 and transformed to E. coli DH5a. The colonies were selected out and cultured, then plasmid was extracted rapidly , digested with enzyme, and expressed under temperature control. Results A recombinant plasmid pBV/IL-2/PEA was constructed. SDS-PAGE showed that the molecular weight of expressed protein was 51 000. Western blot with anti-human IL-2 McAb showed that the expressed product was designed chimeric protein. Conclusion A method for preparing IL-2/PEA chimeric protein was established. The method can also be applied for the expression of other proteins.
2000 01 [Abstract][OnlineView][Download 205k] - Jin Zhigan, Yang Hui, Tian Yuzhen et al (Lanzhou Institute of Biological Products, Lanzhou 730046)
Objective To prepare Haemophilus influenza type b(Hib)polysaccharide protein conju- gate vaccine and study on the immunological characteristics of it. Methods Hib polysaccharide protein conjugate vaccine was prepared by carbodiimide-mediated coupling of adipic acid hydrazide derivtives of Hib capsular polysaccharides with tetanus toxoid(TT). Female NIH mice were subcutaneously injected with 2. 5ug of po- lysaccharide or the conjugate containing 2. 5ug of polysaccharide, and the PRP and TT antibodies in the sera of them were detected by ELISA. Results All the polysaccharide, polysaccharide derivatives and polysaccharide- protein conjugate showed the specific antigenicities of Hib, and the chemical compositions and structures of them were basically consistent with those reported abroad. Single administration of polysaccharide could not induce PRP antibody at a high level in the sera of mice, however, PRP and TT antibodies at high levels were induced in the mice injected with conjugate, and immunological memory was observed. Conclusion The study laid a foundation of further evaluation of protective effect of conjugate.
2000 01 [Abstract][OnlineView][Download 259k] - Yao Zhihui, Dong Guanmu, Yu Yongxin et al (National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050)
Objective To promote the large-scale production of Hantavirus. Methods Hantavirus Z10 strain was adapted in Vero cells. The samples at differenct times were collected and detected for virus titers. Results The strain showed a stable and high titer after being undergone only 2 passages. The peak virus appeared earlier in rolling bottles than that in flasks. At the 4th day after being cultural in rolling bottles, the virus reached a titer of 108/ml and can be harvested for many times. Conclusion The procedure for preparing virus during the production of vaccine has been preliminarily established.
2000 01 [Abstract][OnlineView][Download 122k] -
2000 01 [Abstract][OnlineView][Download 60k] - Zhu Zhiyong, Weng Jingqing, Li Yanjin et al (Zhejiang Health and Antiepidemic Station, Hangzhou 310009)
Objective To improve the immune effect and reduce the adverse reaction of inactivated HFRS vaccine.Methods The concentration effects of stock solution of vaccine by filter membrane with vari- ous sizes were compared by detecting HFRS virus antigen by ELISA. The purification effects of ion exchange chromatography and gel filtration were also compared. Results HFRS virus antigen could he detected out in the stock solution concentrated by the filter membrane with a size of Mr 300 000, but could not be detected out in that concentrated by the filter membrane with a size of Mr 100 000. After the stock solution were purified by Sepharose 4FF gel column chromatography , 3 peaks were observed on the profile at a wavelength of 280nm. However, virus antigen was only detected out in the first peak. Almost all the virus antigens were recovered in the peak, and about 90% of heterologous proteins were removed. Conclusion The purification method is sim- ple, economic and suitable for the large scale production of purified HFRS vaccine.
2000 01 [Abstract][OnlineView][Download 138k] - Yan Hua, Fang Zhizheng et al (Wuhan Institute of Biological Products, Wuhan 430060)
Objective To obtain recombinant HBx-protein with biological acitivity. Methods After expressed inclusion bodies were washed with TNFMX buffer, the HBx-protein in them were degenerated and re- naturated, then purified by affinity chromatography. Results Large amounts of highly purified x-protein were obtained .The purity and recovery rate of the protein reached 96% and 28% respectively. Conclusion The method established by the authors showed good purification effect. It can also be used for the preparation of other recombinant proteins.
2000 01 [Abstract][OnlineView][Download 300k] - Jin Zhezhu, Cai Yingji, Jin Jingling et al (College of Medicine, Yanbian University, Yanji 133000)
Objective To purify the protein with antifungal activity from Aspergillus niger culture flu- id. Methods The protein was seperated by ion exchange column chromatography and further purified by reverse phase HPLC, and the molecular weight of it was determined by tricine gel electrophoresis. Results A kind of protein(AFP) with antifungal activity was seperated. The molecular weight of it was about 9 000. The detection result by MTT method showed significant antifungal activity of the protein to 5 kinds of fungi. Conclusion The study provided a reliable basis for developing drug and antiseptic.
2000 01 [Abstract][OnlineView][Download 202k] -
2000 01 [Abstract][OnlineView][Download 54k] - Huang Xiaoqin, Chen Tongqiu, Tan shunge et al (Institute of Medical Biology, Chinese Academy of Medical Science, Peking Union Medical College, Kunming 650107)
Objective To study on the safety and immunogenicity of inactivated hepatitis A vaccine in common marmosets(Callithris jacchus). Methods After the Lu-8 strain of hepatitis A virus was proliferated in human fetal lung diploid cells(KMB17), the harvested virus was inactivated with formalin and used for inoculating marmosets as an experimental vaccine. Results Specific anti-HAV were induced in the marmosets, and no elevation of serum enzyme activity or pathological exchange of liver tissue were observed. The vaccine protected marmosets against the intravenous challenge with virulent HAV. However, the elevation of serum enz- yme activity and pathological exchange of liver tissue were observed in all the control marmosets. Conclusion The experimental inactivated HA vaccine has good immunogenicity and safety.
2000 01 [Abstract][OnlineView][Download 205k] - Zhang Hongyan, Ling Xiaoyang, Dong Chun' e et al ( Jilin Institute of Family Planning , Changchun 130041)
Objective To prepare a stable and specific diagnostic kit for pregnancy Methods A diagnos- tic kit for pregnancy was prepared by linking of C - terminal polypeptide antibody of anti - HCG - subunit to car- boxyphenylethene latex with bifunctional reagents .Results Latex agglutination test showed that the sensitivity of the kit was 1.5 IU/ml. After being stored at room temperature for 3 months,no decrease of the sensitivity was ob- served. One huned sixty urine specimens from pregnant women and 150 normal urine were detected by the kit. The result showed that the accuracy rate of the kit was 95% .Conclusion The kit is superior to routine kit.
2000 01 [Abstract][OnlineView][Download 135k]
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2000 01 [Abstract][OnlineView][Download 55k] - Liu Songyan,Yu Tingmin, Zhao Li et al (Third Teaching Hospital of Norman Bethune University of Medical Science, Changchun 130031 )
Objective To explore the protective effect of ganglioside on cerebral ischemia/reperfusion. Methods The rat model of cerebral ischemia/reperfusion was established by four-vessel obstruct method. The changes of glutamic acid and γ-aminobutyric acid contents during cerebral ischemia/reperfusion were observed by HPLC,and the denisity of neuron and the influence of ganglioside on the above-mentioned indexes were analyzed by image pattern analyzer.Results The contents of glutamicc acid and aminobutyric acid were significantly increased after cerebral ischemia, and decreased after reperfusionn. Compared with normal saline, ganglioside can significantly decrease glutamic acid content(P <0.01) and increase neuron denisity.Conclusion Ganglioside can ease the cerebral tissue lesion during cerebral ischemia/reperfusion by inhibiting the release of glutamic acid.
2000 01 [Abstract][OnlineView][Download 158k] -
2000 01 [Abstract][OnlineView][Download 277k]
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2000 01 [Abstract][OnlineView][Download 128k] - Chen Weijia, Sui Baozhen, Zheng Hui et al ( Changchun Institute of Biological Products, Changchun 130062)
Objective To observe the curative effect of rIFN-a2a. suppository on chronic inflammation. of pelvic connective tissue. Methods Eighty-three cases of chronic inflammation of pelvic connective tissue complicated by chronic cervicitis were treated by local administration of rIFN-a2a. suppository in cervix uteri, and the HPV-DNA and HCMV-EA in the carvical secretion of a part of the patients were detected before and after treatment. Results The cure, excellent effective, total excellent effective, effective and total effective rates of chronic inflammation of pelvic connective tissue were 59. 7 %, 25. 4%, 85. 1 %, 9. 0% and 94. 1 % respective- ly. The curative effect of rIFN-a2a suppository was significantly superior to that of chlorhexidine suppository. The curative effect on chronic inflammation of pelvic connective tissue was closely correlated with that on chronic cervicitis and the negative conversion of HPV-DNA and HCMV-DNA. Conclusion Local administration of rIFN-a2a suppository in cervix uteri might he effective in chronic inflammation of pelvic connective tissue.
2000 01 [Abstract][OnlineView][Download 174k] - Xu Fugen, Meng Dongmei, Xu Erping et al (Hangzhou Sanitation and Antiepidemic Station, Hangzhou 310006)
Objective To explore if mumps and DT vaccines can be administered simultaneously. Methods The resident students aged 8-14 years were administered with freeze-dried live mumps vaccine and adsorbed purified DT vaccine simultaneously or separately, and the immune responses of them were observed. Results No significant difference were observed in the adverse reactions and immune responses of simultaneous or separate administration of the 2 kinds of vaccine. Conclusion Mumps and DT vaccines can be administered simultaneously.
2000 01 [Abstract][OnlineView][Download 180k] -
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2000 01 [Abstract][OnlineView][Download 174k] 下载本期数据