The partial segment of Marek′s disease virus (MDV) glycoprotein B (gB) gene was amplified by PCR. The segment was cloned into pET-28a vector to obtain the recombinant pET-gB plasmid. The recombinant plasmid was transformed into E.coli BL21,and expressed in very high level as inclusion body after induced with 1.0mmol/L IPTG. The inclusion body was solubilized in urea (8mol/L) . The purified protein was obtained by use of His?Bind affinity chromatography. Mice were immunized i.p. by the purified protein to make the polyclonal antibody. The titer of the antibody by indirect ELISA was 1?10~ -5 . Moreover, the analysis by western blot proved that antibody was specific to the recombinant protein. These works lay a favorable foundation for the study of the immune response by MDV gB.

Objective To compare the ability of the fourth and the third generation HIV assay kits available in Chinese market to detect early HIV infection.Methods 8 BBI HIV seroconversion panels (PRB924,930,940,942,943,944,946 and 948) and 2 National AIDS Reference Lab's HIV seroconversion panels (2004XJ727 and 20505217) were respectively detected with one HIV antigen assay kit,2 fourth generation HIV assay kits and 4 third generation HIV assay kits.The ability of these kits to detect early HIV infection was analyzed and compared.Results For every panel,the fourth generation HIV assay kits could detect HIV-1 infection 4 to 8 days earlier than the third generation kits,and 2 to 4 days later than the antigen kit.The detection ability of different brands of kits was different.Conclusions The fourth generation HIV assay kits could reduce the window period to detect HIV infection.It's meaningful for diagnosing early HIV infection,blood safety and etc.

OBJECTIVETo investigate the current cognition of occupational exposure to human immunodeficiency virus (HIV) and the personal occupational protection awareness in healthcare workers in Liuzhou, China.

METHODSA total of 270 healthcare workers were selected from 10 hospitals in Liuzhou by stratified random sampling for a cross-sectional study. And a self-administered questionnaire of occupational exposure to HIV was designed to conduct a survey. The descriptive analysis of data was carried out by Excel. And a logistic regression analysis was done to analyze the effects of different factors on healthcare workers' cognition of occupational exposure to HIV using the statistical analysis software SPSS 19.0.

RESULTSA total of 260 usable questionnaires (96.3%) were returned. Among them, 220 healthcare workers (84.6%) had received the trainings on occupational exposure to HIV; 223 healthcare wofkers (85.8%) were aware of the rules and regulations on prevention of occupational exposure to HIV and the operation procedures in their hospitals. The healthcare workers who had not received the trainings or had not known the rules and regulations and the operation procedures were mainly from primary or secondary hospitals. A total of 106 healthcare workers (40.8%) had directly contacted patients' blood or body fluids; 154 healthcare workers (59.2%) were injured by sharp instruments, and most were hollow needle stick injuries (98/154, 63.6%). A total of 168 healthcare workers (68.08%) had better cognitive awareness of occupational exposure to HIV, and 76 healthcare workers (29.2%) had moderate cognitive awareness. Some healthcare workers had poor knowledge in the common sense of AIDS/HIV and occupational exposure to HIV, the personal protection awareness of occupational exposure, or the disposal measures after occupational exposure. The univariate analysis using chi-square test showed that occupation and professional title were significantly correlated with the cognition (P<0.05). The multivariate logistic regression analysis showed that the doctors (OR3.8; P<0.05), nurses (OR3.04, P<0.05), and laboratory technicians (OR=9.51, P<0.05) had better awareness compared with the others. The healthcare workers with a primary or lower professional title had poorer awareness compared with the healthcare workers with a higher professional title (OR=0.47, P<0.05).

CONCLUSIONHealthcare workers have the risk of occupational exposure to HIT. They do not have comprehensive and systematic knowledge related to occupational exposure to HIV, and they have a high demand for training.

Awareness ; China ; Cross-Sectional Studies ; HIV Infections ; prevention & control ; Health Knowledge, Attitudes, Practice ; Health Personnel ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; prevention & control ; Laboratory Personnel ; Needlestick Injuries ; Nurses ; Occupational Exposure ; prevention & control ; Physicians ; Risk ; Surveys and Questionnaires

The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.

OBJECTIVETo investigate the protective effect of recombinant human N-terminal lipopolysaccharide binding protein in mice challenged with LPS.

METHODSSeventy male Kunming mice were randomly divided into 3 groups, i.e. LPS challenge (Injection of LPS into abdominal cavity, n = 21); tLBP protection (Injection of LPS and tLBP into abdominal cavity, n = 21) and control (Injection of normal saline into abdominal cavity, n = 8) groups. The blood samples and tissue samples of the liver and lungs were harvested on 15 and 30 minutes and 1, 3, 6, 12 and 24 hours after the injection. The serum contents of ALT and TNF-alpha were determined by biochemical velocity analysis and RIA method, respectively. The pathomorphological changes in the liver and pulmonary tissue were examined under light microscope (LM). The mortality rate of ten mice each was observed within 24 hours after the injection of tLBP + 400 ng LPS or 400ng LPS.

RESULTSThe ALT content of tLBP group reached the peak level at 12 post-injection hour (PIH) (41.00 +/- 4.58), but it was significantly lower than that in LPS group in which it peaked at 6PIH (99.50 +/- 62.63) (P < 0.01). The TNF-alpha content in tLBP and LPS group was lower than that in LPS group, and both reached the peak level at 3 PIH (35.96 +/- 7.33). Compared with those in LPS, injury to hepatocytes in tLBP group was obviously milder without scattered necrosis. The pulmonary congestion in tLBP group was abated, and the inflammatory exudation in the alveoli was evidently less than that in LPS group. There were 9 out of 10 mice died in the LPS challenge group, while only 3 out of 10 mice died during 24 hours after LPS injection in tLBP protection group.

CONCLUSIONPreliminary results indicated that recombinant human tLBP might possess biological activity with a potential protection effect in LPS challenged mice.

Acute-Phase Proteins ; Animals ; Carrier Proteins ; therapeutic use ; Lipopolysaccharides ; toxicity ; Liver ; drug effects ; pathology ; Lung ; drug effects ; pathology ; Male ; Membrane Glycoproteins ; Mice ; Random Allocation ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo review the presentations of intracranial venous sinus lesions in phase-contrast (PC) magnetic resonance angiography (MRA) and discuss the diagnostic value of this imaging modality for these lesions.

METHODSThis study involved 52 patients with intracranial venous sinus lesions, including 21 with dural venous sinus malformation, 7 dural arteriovenous fistula (6 caroticocaveneous fistula, CCF), 5 venous sinus thrombus and 19 meningioma invading the venous sinuses. All patients underwent PC and time-of-flight (TOF) MRA with a 1.5 Tesla MR scanner, with 28 of them undergoing subsequent digital subtraction angiography (DSA).

RESULTSPC MRA showed extensive hyperintense signals that identified the lesions, feeding arteries, deferent veins and venous sinuses in 21 cases of dural arteriovenous fistula, but the lesion signals appeared homogeneous. In these cases, PC and TOF MRA had equally good performance in displaying the feeding arteries with success in 20 of the 21 cases (95.2%), whereas the former resulted in a greater rate of deferent vein display [90.5% (19/21)] than the latter [76.2% (16/21), P<0.05]. The 6 CCF cases showed ipsilateral cavernous sinus expansion and upper ophthalmic vein dilations. In the 5 cases of venous sinus thrombis, 4 had transverse sinus thrombus and 1 superior sagittal sinus thrombus. All the cases presented signal disappearance in the sinuses in PC MRA and hyperintense signals in MRI, and some cases had also cerebral superficial hemorrhage and edema. The 19 cases of meningioma invading the venous sinus displayed sinus shift, stenosis and occlusion in PC MRA.

CONCLUSIONPC MRA has great advantages in displaying the anatomical structure and lesions of the venous sinuses.

Adolescent ; Adult ; Aged ; Central Nervous System Vascular Malformations ; diagnosis ; Child ; Cranial Sinuses ; pathology ; Female ; Humans ; Magnetic Resonance Angiography ; methods ; Male ; Meningeal Neoplasms ; diagnosis ; Meningioma ; diagnosis ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity

Objective:To investigate the expression of interleukin-1 receptor associated kinase-M(IRAKM)in systemic lupus erythematosus ( SLE) and its relationship with immunoregulation .Methods:103 patients with SLE were divided into active stage group (n=55) and stable stage group(n=48) according to their disease activity score (SLEDAI),and 40 healthy persons were chosen as control group.Real-time quantitative PCR ( RT-PCR ) was used to detect the expression of IRAKM mRNA in peripheral blood monouclear cells of the three groups .The levels of anti ds-DNA antibody and anti Sm antibody in serum were detected by Enzyme linked immunosorbent assay(ELISA).The levels of serum complement C3 and C4 were measured by immune scattering turbidimetry .Factor analysis of variance was performed to analyze the differences of all the observed indicators in the three groups .The correlation between IRAKM and autoantibodies and complements of SLE was analyzed by Pearson or Spearman .Results: The expression level of IRAKM mRNA of SLE patients in the active stage group and stable stage group were significantly lower than that in the control group ( P<0.05).Moreover,the expression level of IRAKM mRNA in the active stage group was lower than that in the stable stage group ,and the difference was statistically significant ( P<0.05 ) .Compared with the control group , the levels of anti ds-DNA antibody and anti Sm antibody in the active stage group and stable stage group were significantly increased ( P<0.05 ) ,and the levels of complement C 3 and C4 were obviously lower(P<0.05).The changes of serum levels of autoantibodies and complements in the active stage group were more remarkable than those in the stable stage group ( P<0.05 ) .The expression level of IRAKM mRNA in the SLE patients was negatively correlated with the levels of anti ds-DNA antibody and anti Sm antibody (P<0.05).However,it was positively correlated with the levels of complement C3 and C4 ( P<0.05 ) .Conclusion: IRAKM participates in the immune regulation process of SLE through negative regulation,and its expression level is closely related to the degree of SLE disease activity .

The risk factors of high trait anger of juvenile offenders were explored through question naire study in a youth correctional facility of Hubei province,China.A total of 1090 juvenile offenders in Hubei province were investigated by self-compiled social-demographic questionnaire,Childhood Trauma Questionnaire (CTQ),and State-Trait Anger Expression Inventory-Ⅱ (STAXI-Ⅱ).The risk factors were analyzed by chi-square tests,correlation analysis,and binary logistic regression analysis with SPSS 19.0.A total of 1082 copies of valid questionnaires were collected.High trait anger group (n=316) was defined as those who scored in the upper 27th percentile of STAXI-Ⅱ trait anger scale (TAS),and the rest were defined as low trait anger group (n=766).The risk factors associated with high level of trait anger included:childhood emotional abuse,childhood sexual abuse,step family,frequent drug abuse,and frequent internet using (P＜0.05 or P＜0.01).Birth sequence,number of sibling,ranking in the family,identity of the main care-taker,the education level of care-taker,educational style of care-taker,family income,relationship between parents,social atmosphere of local area,frequent drinking,and frequent smoking did not predict to high level of trait anger (P＞0.05).It was suggested that traumatic experience in childhood and unhealthy life style may significantly increase the level of trait anger in adulthood.The risk factors of high trait anger and their effects should be taken into consideration seriously.

Objective To investigate the status of genotype of the KPC(Klebsiella pneumoniae carbapenemase)-encoding genes in Pan-resistant K. Pneumoniae, isolated from the 98th Hospital of People' s Liberation Army, Huzhou district, Zhejiang province, China. Methods 19 strains of Pan-resistant K. Pneumoniae were isolated from the inpatients between November, 2008 and July,2009. Phenotypic confirmatory test for suspected carbapenemases production were carried out by Modified Hodge test. Carbapenemase gene of blaKPC was analyzed by PCR and verified by DNA sequencing. Results In 19 strains of K. Pneumoniae, the positive rates of Modified Hodge test and gene of blaKPC were both 100.0%. These genes all belonged to blaKPC-2 subtype confirmed by nucleotide sequence analysis. Among them, the blaKPC-2 gene sequence of the HZ001 strain (its original serial number was HZ9871 ) had been registered in GenBank (GenBank Accession Number: GU086225).Conclusion All of the Pan-resistant K. Pneumoniae isolated from the inpatients harbored blaKPC-2 type carbapenemases gene and causing an outbreak in a hospital. Carbapenemases that producing type KPC-2 might be the major reason which causing the resistance to Carbapenems antibiotics.

Objective To study the effect of atorvastatin on the expressions of intercellular adhesion molecule-1 (ICAM-1) and transforming growth factor-β1 (TGF-β1) in the spinal cord of rats with experimental autoimmune encephalomyelitis (EAE).Methods Eighty healthy female Wistar rats were randomly divided into 4 groups:normal control group (n=20),EAE model group (n=20),low-dose treatment group (n=20) and high-dose treatment group (n=20).Each group was equally divided into 2 sub-groups:14-d group and 21-d group according to the day that they were sacrificed.EAE rat models were established by immunizing the rats with fresh guinea-pig spinal cord homogenate plus complete Freund's Adjuvat (CFA).Rats of the treatment groups were fed with atorvastatin at dasage of 2 mg/(kg·d)and 10 mg/ (kg·d),respectively; and rats of the normal control group and EAE model group were fed with physiological saline.The severity of EAE was scored according to the signs and symptoms,and NIHSS was performed.Pathological changes were observed with the aid of HE staining.The levels of ICAM-1 and TGF-β1 were detected by immunohistochemistry.Results The rats in the high-dose treatment group had significantly lower incidence of disease,decreased CNS inflammation focus,lower level of ICAM-1 and higher level of TGF-β1 as compared with rats of the EAE model group and low-dose treatment group (P＜0.05).Conclusion Atorvastatin could ameliorate EAE of rats in a dose-dependent manner,whose effect might be related to the decreased expression of ICAM-1 and increased expression of TGF-β1.