2.Prokaryotic expression of HN gene of bovine parainfluenza virus type 3 and the establishment of indirect ELISA method.
Yu-Long ZHOU ; Ya-Chao REN ; Zhan-Bo ZHU ; Xi-Lin HOU ; Mi WANG ; Jing GENG ; Fan-Ze PIAO ; Sen LI
Chinese Journal of Virology 2012;28(1):23-28
The prokaryotic expression plasmid pQE30-HN of hemagglutinin-neuraminidase (HN) protein gene of bovine parainfluenza virus type 3 (BPIV3) strain HJ-1 was expressed by IPTG induction in E. coli XL1Blue. The recombinant HN protein(rHN) was purified by electroeluting method, and used as coated antigen. An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect the antibody valence of BPIV3. The best working conditions of ELISA were as follows: the antigen concentration was 6 microg/mL; the serum dilution was 1:50; the blocking reagent was 5% skimmed milk; the blocking time was 60 min at 37 degrees C; the second antibody concentration was 1:10 000; The cut-off value was 0.30. The method revealed a good specificity, no cross-reaction to the positive sera of BCV, IBRV or BRSV was observed. We applied the method to detect 323 serum samples of dairy cow in Heilongjiang Province, the seropositivity rate of BPIV3 was about 58%. The indirect ELISA established provided a technological basis for the development of ELISA kit.
Animals
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Antibodies, Viral
;
blood
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Blotting, Western
;
Cattle
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Enzyme-Linked Immunosorbent Assay
;
methods
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Escherichia coli
;
genetics
;
Female
;
HN Protein
;
genetics
;
Parainfluenza Virus 3, Bovine
;
genetics
;
immunology
3.Ultrastructural changes of the rat convoluted seminiferous tubule-after alcohol consumption.
Ting JIN ; Yong-jiu ZHAO ; Zhao-dian CHEN ; Li-ming SHEN ; Chao-hui ZHAN ; Fu-jin REN ; Jie YAO ; Zhong-mu TANG ; Xiao-feng ZHU
National Journal of Andrology 2006;12(6):505-509
OBJECTIVETo study the ultrastructural changes of the rat convoluted seminiferous tubule after alcohol consumption.
METHODSForty-eight Wistar mature male rats were divided into two groups randomly: control group (A) and experimental one (B). 6 ml/(kg x d) of 50 degrees alcohol was perfused through the gastric tube for 39 days in Group B; and 6 ml/(kg x d) of normal saline was supplemented in Group A. The ultrastructure of the rat convoluted seminiferous tubule was observed by transmission electron microscope at day 14, 27 and 40.
RESULTSIn Group A, the pykno-basement membrane was unstriated and uniform, Sertoli cells showed cytoplasmic profusion, with big nucleus, well-distributed nucleoplasm, distinct nucleolus, more mitochondria and plain hierarchical tight-junction. And the ultrastructure of the rat convoluted seminiferous tubule in Group B began to change at the end of the first spermatogenic cycle (D 14) and changed more and more evidently with the ethanol administration, mainly as follows: (1) more lysosomes and vacuolisation found in Sertoli cells, and organelles decreased and blurry; (2) more and bigger vacuoles among the spermatogonia, Sertoli cells and basement membrane; (3) obvious apoptosis of spermatogonia and apoptotic bodies aggregated near the membrane; (4) more cytoplasm and vacuolisation in the sperm of the convoluted seminiferous tubule, and disarranged, deleted or clustered mitochondria in the sperm tail; (5) blurry and rigid tight-junction; (6) thickened, wrinkled or broken basement membrane and under-basement
CONCLUSIONAlcohol can cause ultrastructural changes of the basement membrane, tight-junction and Sertoli cells of the membrane. rat convoluted seminiferous tubule and apoptosis of spermatogonia.
Animals ; Apoptosis ; drug effects ; Basement Membrane ; drug effects ; pathology ; Ethanol ; toxicity ; Male ; Microscopy, Electron, Transmission ; Random Allocation ; Rats ; Rats, Wistar ; Seminiferous Tubules ; drug effects ; ultrastructure ; Sertoli Cells ; drug effects ; pathology