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.

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.

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

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 explore the expression and distribution of transient receptor potential cation channel 6(TRPC6)in normal human,mice,rats' renal tissue and the mouse podocyte clone 5(MPC5)for further investigating the relationship between TRPC6 and the protei-nuria-related podocyte molecules.Methods 1.The distributions of TRPC6 in normal human,mice,rats' renal tissue and MPC5 were observed by using the immunochemistry staining.2.The mRNA expression of TRPC6 in mouse renal cortex and differentiated MPC5 was detected by using reverse transcriptase-polymerase chain reaction(RT-PCR).3.The protein expression of TRPC6 in human,mice and differentiated MPC5 was detected by using Western blotting.Results 1.In human kidney,TRPC6 showed a weak staining in glomeruli and a strong staining in renal tubules and vessels.In mice and rats' kidney,TRPC6 showed a strong staining in glomeruli and was mainly distributed along the capillary loops of glomerulus and in mesangium.The positive staining of TRPC6 was observed in MPC5,which was distributed evenly on the cell membrane in differentiated podocytes.2.The specific PCR band of TRPC6 was detected in mouse renal cortex and differentiated MPC5.3.The specific protein band of TRPC6 was detected in normal human,mice renal cortex and differentiated MPC5 with the size of 106.Conclusions The expression of TRPC6 is verified in normal human,mice and rats' kidneys,and in differentiated MPC5.These results will benefit for further exploring the relationship between TRPC6 and the proteinuria-related podocyte molecules.

Objective To explore the value of deoxyribonucleic acid(DNA) content on clinical pathological characteristics of nephroblastoma in children.Methods Twenty-four cases of pediatric nephroblastoma from Jan.1993 to Dec.2007 were proved by pathology.The distribution of the 24 cases according to National Wilms Tumor Study group(NWTSG) criteria of USA was stage Ⅰ(12 cases),stage Ⅱ(7 cases),stage Ⅲ(5 cases).The pathology was highly differentiated in 10 cases,medium differentiated in 8 cases and low differentiated in 6 cases according to the level of differentiation.There were 9 cases of lymphatic metastasis and 15 cases of non-lymphatic metastasis in accordance with lymphatic metastasis.The DNA contents of 24 cases were determined by flow cytometry.Chi-square test was used to analyze the relationship among DNA ploidy and clinical stage,level of differentiation,lymphatic metastasis of children with nephroblastoma.Results Aneuploidy mainly occurred in clinical stage Ⅱ or stage Ⅲ(57.14%,100.0%),there was significant difference compared with that of stageⅠ(25.0%)(P