Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Cause of Death ; Humans ; Male ; Middle Aged ; Mining ; Pneumoconiosis ; mortality ; Retrospective Studies

China ; Foundations ; Preventive Medicine ; economics ; Research Support as Topic ; statistics & numerical data

Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.

Objective To investigate the serum adiponectin levels and TNF-? levels in women with gestational diabetes and to study their clinical significance.Methods Plasma concentration of adiponectin,tumor necrosis factor alpha and correlate parameters were measured in 50 patients with gestational diabetes(GDM)and 36 pregnant women with normal glucose tolerance(NGT).Radio immunoassay was used to measure the adiponectin,TNF-? level was measured by ELISA.Serum insulin level was measured by Electrochemiluminescent immunoassays fasting glucose levels by glucose oxidase method.Results GDM patients had significantly lower concentrations of adiponectin(10.3?(2.4) mg/L) and elevated levels of TNF-?(4.6?(1.5)?g/L) in comparison to NGT women.In GDM group TNF-? level was correlated positively with serum insulin level,glucose level.Concentration of adiponectin was negatively correlated with corresponding parameters.Conclusion Elevated level of TNF-? and decreased adiponectin concentration may not simply reflect maternal adiposity and insulin resistant state,but may contribute to the impaired glucose metabolism during pregnancy and forecast the risk of metabolism syndrome.

0.05) at any observed time points as compared with the controls.3.From day 7 after the adriamycin injection,VEGF protein reduced significantly(P

Objective To investigate the prevalence features of helicobacter pylori (Hp) infection,anemia and iron deficiency in a po-pulation of Wuhan children with 2 to 6 years old,and the relationship between Hp infection and anemia,iron deficiency in the children.Methods Randomly taking 95 children who had taken tests in our hospital's check-up centre in 2008 as the study objects,2 kinds of exa-mination were employed to detect Hp infection.Serum levels of Hp-IgG were measured by enzyme linked immunosorbent assay (ELISA) methods to evaluate past infection.The 14C urea breath test (14C-UBT) was conducted to obtain information of the presence of current/active Hp infection.In the morning 3 mL fasting venous blood was collected to determine the serum levels of Hp-IgG antibodies and ferritin.Hemoglobin values were determined with a hemoglobinometer.Serum levels of C-reactive protein (CRP) were tested in order to determine whether the children had evidence of current inflammation or infection.In addition,demographic information such as age and gender of the children and information about their use of antibiotics within the prior month were recorded.All cases were divided into 2 groups including the Hp infection group and non-Hp infection group according to laboratory examinations,then the Logistic regression was applied to analyze the relationship between Hp infection and anemia,iron deficiency.The Kappa identity test was taked to compare the 2 measures.Results Of the 95 children,18.9% were anemic and 36.8% were iron deficient.Forty percent of the cohort had Hp-IgG antibodies,74.4% tested positive by the UBT.Presence of Hp-IgG emerged as a significant risk factor for anemia,iron deficiency in adjusted analysis controlling for demographic factors,current inflammation,and antibiotic use.Conclusions Findings from different measure of Hp may reflect different stages of infection,with UBT results reflecting an earlier stage of infection,and presence of Hp-IgG reflecting established Hp infection associated with anemia,iron deficiency.

ObjectiveTo determine the potential natural foci of new bunyavirus,and isolate and identify the new bunyavirus strain in sera from suspected new bunyavirus-infected patients.MethodsImmunofluorescence assay was used to detect the antigens of new bunyavirus in different tissue specimens of wild rodent animals in Tiantai area of Zhejiang province.Fluorescence quantitative real-time RT-PCR was applied to detect the viral nucleic acid in sera of suspected new bunyavirus-infected patients and the amplification products were analyzed by sequencing.The new bunyavirus in the pateints'sera was isolated using Vero cells.Using nucleocapsid protein encoding gene of new bunyavirus as the target gene,the isolated suspected new bunyavirus strain was identified by RT-PCR and sequencing of the amplification product.Moreover,sequence identity of the amplification product of nucleocapsid protein encoding gene of new bunyavirus was analyzed and compared.ResultsOf the 70 wild rodent animals,5.71% were positive in the immunofluorescence assay.The fluorescence quantitative real-time RT-PCR confirmed that two of the four detected patients'serum specimens were positive.One suspected strain of new bunyavirus was isolated from one pf the two positive patients'serum specimens.The results of RT-PCR and sequencing confirmed that the viral strain exactly belongs to new bunyavirus with 92.2% sequence identity to that of the new bunyavirus isolates in Hubei province but distinct with the new bunyavirus isolates from other areas in China.ConclusionThe presence of natural foci of new bunyavirus and new bunyavirus-infected patients in Zhejiang province are firstly confirmed by this study.There is a geographical diversity of the distribution of new bunyavirus in different groups.

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.

Objective To observe the changes of inflammatory factors and clinical parameters on septic patients with hemoperfusion,and to discuss the application of hemoperfusion on sepsis. Methods 43 patients with sepsis were divided into treatment group and control group randomly. In the treatment group,the patients received conventional treatment and hemoperfusion together,which performed every 24 hours,continuously for 3 times when they arrived in ICU in the first hour. The concentrations of TNF-α,IL-6,IL-10 and PAF were dynamically detected before hemoperfusion,after 24 hours,48 hours and 72 hours in treatment group. The concentrations of TNF-α,IL-6,IL-10 and PAF were compared between the two groups after 72 hours. So did the clinical parameters as WBC count,CRP,PCT and blood lactate acid. Results The concentrations of TNF-α,IL-6, IL-10 and PAF were increased significantly in the early stage of sepsis,and were decreased obviously after hemoperfusion(P < 0.01). After 72 hours treatment,the concentrations of TNF-α,IL-6 and PAF were decreased rapidly,so did the level of CRP,PCT and blood lactate acid. There were significant differences between the two groups(P < 0.05). ConclusionHemoperfusion could remove the inflammatory factor of septic patients and improve the clinical symptoms of them.

Purpose To investigate the feasibility to diagnose and evaluate the vulnerability of the atherosclerotic plaque through contrast-enhanced ultrasonography. Materials and Methods Thirty-four patients with carotid atherosclerosis were enrolled in the study to take contrast-enhanced ultrasonography and were observed on whether the plaque was enhanced and the features of its enhancement. The peak intensity (PI), time to peak (TTP), and density echo (DE) were calculated according to the time-intensity curve with QLAB software. Results The ratio of enhanced malacoplakia and the mixed plaque showed no difference (χ2=0.847, P>0.05). The percentages of enhanced plaque distributed on the base, tail and the shoulder were 70.0%, 23.3% and 6.7%, respectively with significant difference (χ2=29.100, P<0.001); the distribution ratio of enhanced plaque from high to low were as follows: the plaque on the shoulder > the plaque on the base> the plaque on the tail. It was positively correlated between enhanced plaque and its distribution (r=0.404, P<0.01). The TTP of the ROI between the malacoplakias and the mixed plaques showed no difference (t=0.479, P>0.05). The PI and DE of the ROI in the malacoplakia and the mixed plaques were analyzed by the time-intensity curve and the differences proved to be statistically significant (t=7.497 and 12.224, P<0.05). Conclusion Contrast-enhanced ultrasonography could present the neovessels in the atherosclerotic plaque, which is helpful in evaluating the vulnerability of atherosclerotic plaque.