1.Examination of A.hydrophila Isolated from Whitmania pigra (L.)
Microbiology 1992;0(01):-
We examined diseases occurring farmed Whitmania pigra(L.) in Hebei province in the aspect of situation of disease,clinical symptoms and pathological changes.In addition,the molecular identification were conducted to representative strain,the 16S rRNA gene was sequenced and compared with that of related strains,molecular phylogenetic tree was constructed.The results showed that the disease was infected by Aeromonas hydrophila.Pure cultures of 10 strains have the same serotype.Selected representative strain was proved to be the corresponding primitive causal agent of the disease by artificial infection experiment to healthy Whitmania pigra(L.).Antibiotic sensitivity of the isolates to used thirty-seven antimicrobial agents showed that the tested strains were high sensitive to cefotaxime et al.,were sensitive to streptomycin et al.,were resistant to oxacillin et al.
3.Identification of Acinetobacter junii Morphovari from Diseased Stone Flounder (Kareius bicoloratus L.)
Cui-Zhen CHEN ; Hai FANG ; Xiao-Jun ZHANG ;
Microbiology 1992;0(03):-
Successive pathogenic bacteria were examined from liver,spleen, kidney and contents of intestine of diseased stone flounder (Kareius bicoloratus L.) occurring bacterial septicaemia infection caused by Aeromonas salmonicida. Identification of extensive phenotypic information to 6 pure cultures and detection of the mol% G+C ratio of the DNA to representative strains,showed that the examined bacteria belonged to a new morphovar of Acinetobacter junii, and was designated as Acinetobacter junii morphovar I. In addition, serotype, antibiotic sensitivity and pathogenicity of isolates were studied, the results showed that the 6 strains have the same K-antigen and O-antigen, there are no obvious differences in sensitivity and resistance to used 37 antimicrobial agents between strains, and have strong pathogenicity to experimental stone flounder and Bastard halibut.
4.Expression of Truncated NSP2 Protein of Porcine Reproduction and Respiratory Syndrome Virus in E.coli and Preparation of Monoclonal Antibodies Against NSP2 Protein
Hai-Yan WANG ; Ping JIANG ; Yi-Jun DU ; Yu-Feng LI ; Jun-Xing LI ; Fang SHEN ;
China Biotechnology 2006;0(02):-
The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.
5.Clinical study on the severe hepatitis with nosocomial fungal infections and risk factors.
Xue-Hai ZHANG ; Guang-Hai ZHANG ; Chang-Jun MAN ; Fang-Ming HE
Chinese Journal of Hepatology 2004;12(7):389-391
OBJECTIVETo study the nosocomial fungal infections in the patient with severe hepatitis and analyze of risk factor.
METHODSAll 115 severe hepatitis with fungal infections inpatients was studied prospectively.
RESULTSWe identified 115 cases with fungal infections, the mean age of patients was 37.2+/-21.5 years, male: 49 cases, female 66 cases. Infection of abdominal cavity accounted for 40.9%, infectious rate in respiratory tract and digestive tract were 26.9%, 21.8%, respectively. Candida albicans accounted for 67.6%. Use of broad-spectrum antibiotic and corticosteroids, neutropenia, severity of liver disease, improper medical manipulations as significant risk factors for fungal infection. Death rate of study group and control group was 59.1%, 34.8%, respectively (x2=36.0). In multivariate analysis, neutropenia, disseminated infection and severity of liver diseases were independent prognostic factors.
CONCLUSIONIdentification of risk factors and predictors of a poor outcome in patients with severe hepatitis with fungal infections, it suggested that implications in prophylaxis of fungal infection, early diagnosis and appropriate therapy would be important for these patients.
Adult ; Candidiasis ; diagnosis ; epidemiology ; China ; epidemiology ; Cross Infection ; complications ; epidemiology ; Female ; Hepatitis, Viral, Human ; complications ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Mycoses ; epidemiology ; microbiology ; Prospective Studies ; Risk Factors ; Severity of Illness Index
6.Expression of NSP 3AB Gene of Encephalomyocarditis Virus(EMCV)in E.coli and Development of Monoclonal Antibodies Against 3AB Protein
Fang SHEN ; Ping JIANG ; Yu-Feng LI ; Jun-Xing LI ; Hai-Yan WANG ;
China Biotechnology 2006;0(09):-
Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.
7.Inhibitory effect of 5-fluorouracil encapsulated by galactosylceramide liposomes on drug resistance and its mechanisms
Yong JIN ; Jun LI ; Yuan-Hai LI ; Xiong-Wen LV ; Jin-Fang GE ; Shu-Yun XU ;
Chinese Pharmacological Bulletin 2003;0(08):-
Aim To study inhibitory effect of 5-fluorouracil encapsulated by galactosylceramide liposomes (5-Fu-GCL)on 5-Fu-resistent HepG_2 cells and its mechanisms. Methods Inhibitory effect of 5-Fu-GCL on established model of 5-Fu-resistant HepG_2 cells was assessed with MTT assay in vitro. The concentration-time course of 5-Fu-GCL in intracellular fluid was detected with high performance liquid chromatography (HPLC). Thymidylic acid synthase (TS) expression was observed with immunohistochemical method,and NO content was determined with chemical method. Results Obvious inhibitory effects of 5-Fu-GCL (75,150,300,600,1200?mol?L-1) on 5-Fu-resistant HepG_2 cells were observed with IC_ 50 of 158.6 ?mol?L-1,far lower than that of free 5-Fu (400.9 ?mol?L-1). 5-Fu-GCL (300 ?mol?L-1) inhibited 5-Fu-resistant HepG_2 cells in a time-dependent manner,and the inhibitory effect of 5-Fu-GCL was stronger than that of free 5-Fu during 12~48 h. Compared with free 5-Fu,5-Fu-GCL (300 ?mol?L-1) increased the content of intracellular fluid in 5-Fu-resistant HepG_2 cells. 5-Fu-GCL(62.5,300,1200 ?mol?L-1) not only inhibited the expression of TS,but also increased the production of NO in 5-Fu-resistant HepG_2 cells,and these effects of 5-Fu-GCL(300,1200 ?mol?L-1) were stronger than those of free 5-Fu. Conclusion 5-Fu-GCL has inhibitory effect on 5-Fu-resistant HepG_2 cells. The effect may be related to the increased concentration of 5-Fu-GCL in intracellular fluid,inhibited expression of TS and increased production of NO.
8.Effect of nerve growth factor on denervated bone fracture healing in rats
Cheng MA ; San-Huai GOU ; Hai-Jun XIAO ; Yue-Ping OUYANG ; Yan LIU ; Fang HE ;
Academic Journal of Second Military Medical University 1985;0(05):-
Objective:To study the effect of nerve growth factor(NGF)on bone fracture healing of inflicted T_(10)spinal cord injury(SCI)complicated with tibia fracture in rats.Methods:Totally 120 rats were randomly divided into tihia fracture group (F group,n=40),T_(10)SCI+tibia fracture group(FS group,n=40),and T_(10)SCI+tibia fracture+NGF group(FSN group,n=40).Four weeks later,the fracture sites in the 3 groups were subjected to CT scanning;the maximum transverse diameter of the fracture ends and the gray scales of non-osseous area were measured;the changes of biomechanics property of the fracture ends were determined by three-point bending test;the bone morphometry,bone density,and histomorphology of callus were determined;the expression of OCN was detected by immunohistochemical method;the osteoblast ultrastructure was observed by TEM and the expression ofⅠ,Ⅱtype collagen were examined by Western blotting.Results:The maximum transverse diameter of F group was less than those of FS group(P
9.Clinical analysis of Staphylococcus aureus resistance to methicillin in patients with coal worker's pneumoconiosis complicated by lung cancer.
Si-hai LIU ; Pei-yue LIU ; Wen FENG ; Jun-he DAI ; Cheng-dong QI ; Fang QIAN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(5):391-392
10.Evaluation of Urodynamic Study and Somatosensory Evoked Potentials before and after Operation in Children with Lipomyelomeningocele
zhe, GAO ; hai-tao, GU ; gang, WANG ; jun-ping, HE ; yun-fei, GUO ; fang, LIU
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To evaluate prourodynamic study and somatosensory evoked potentials(SEPs)before and after operation of children with lipomyelomeningocele(LMMC)and its clinical significance.Methods Urodynamic study(UDS)and SEPs in 31 cases of LMMC who underwent microsurgical release within 1 week preoperatively and 3 months postoperatively were conducted.The 4 parameters used for UDS evaluation were bladder volume,compliance,detrusor activity and residual urine;parameters used for SEPs evaluation were latent period and amplitude.Results There was a statistics significant difference between operation and control group;the similar result was attented in the comparison of before and after operation,also in the comprison of the improved group and the group without improvement after operation.Conclusions UDS and SEPs investigation can provide guidance for the treatment of tethered cord syndrome(TCS)and evaluate clinical prognosis.These cases with the severity changing in UDS and somatosensory shall be avoided unnecessary action in operation.