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.

Acute Disease ; Adolescent ; Adult ; Electromyography ; Female ; Humans ; Male ; Middle Aged ; Nervous System Diseases ; chemically induced ; diagnosis ; Organophosphate Poisoning ; Pesticides ; poisoning

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.

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.

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

The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.
Biological Transport ; Caco-2 Cells ; Chitosan ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Coculture Techniques ; Drug Carriers ; Furans ; administration & dosage ; chemistry ; metabolism ; HT29 Cells ; Heterocyclic Compounds, 2-Ring ; administration & dosage ; chemistry ; metabolism ; Humans ; Lactic Acid ; chemistry ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglycolic Acid ; chemistry ; Zonula Occludens-1 Protein ; metabolism

Adult ; Aged ; Aged, 80 and over ; Coal Mining ; Humans ; Lung Neoplasms ; complications ; microbiology ; Male ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; complications ; microbiology ; Staphylococcus aureus ; drug effects ; isolation & purification

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.

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

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.