Relationship between urinary albumin-to-creatinine ratio and prehypertension among young migrant builders in Shanghai / 第二军医大学学报

Objective To explorethe relationship between urinary albumin-to-creatinine ratio (UACR) and blood pressure of young migrant builders in Shanghai. Methods A total of 3 195 builders (age ranged from 18 to 50) were selected from migrant builders of Shanghai Construction Group Co., Ltd. by using random clustersampling method. They were subjected to questionnaire interview, physical examination, and biochemical tests, including UACR, renal function, serum uric acid, serum lipid, and serum glucose. The participants were divided into 4 groups (I, II, III, and IV) according to the levels of UACR, and the ordinal multinomial logistic regression model was employed todetermine the relationship between UACR levels and blood pressure grouping (normal, pre-hypertension, and hypertension). Results Among the 3 195 builders, 3 112 (97. 4%) finished interview. The UACR (median [Q1-Q3]) levels and high-UACR ratio for the normal, pre-hypertension and hypertension groups were 0. 8 (0. 6-1. 2) mg/mmol, 0. 9 (0.7-1. 4) mg/mmol, and 1. 0 (0.8-1. 7) mg/mmol, and 6. 8%, 8. 0%, and 15. 6 %, respectively, with significant differences for both UACR levels and high-UARC ratios (P<0. 05) among the 3 blood pressure groups. According to the 25%, 50%, and 75% quartiles of UACR values, builders were classified into 4 groups. After adjustment of the other factors such as age, the average systolic pressures of the I, II, III, and IV UACR groups were (126. 8± 13. 3), (127. 9 ± 14. 2), (129.9 ± 14. 4), and(134. 2 ± 19. 0) mmHg (1 mmHg= 0. 133 kPa), and the average diastolic pressures of the 4 groups were (77. 6±9. 7), (78. 8±9. 2), (79. 2 ± 10. 4), and (81. 8 ± 12. 5) mmHg, respectively. UACR was positively correlated with both the average systolic pressures and diastolic pressures (P<0. 05). Logistic regression analysis showed that, after adjusting the factors such as age, body mass index, total cholesterol, and low density lipoprotein cholesterol, the odds ratio (OR) and 95% confidence interval (95% Cl) of pre-hypertension and/or hypertension, compared with I group, were 0. 98 (0. 80-1. 20), 1. 29 (1. 05-1. 58), and 1. 86 (1. 52-2. 28) for II, III, and IV groups, respectively. Conclusion The UACR is positively correlated with pre-hypertension and hypertension among young migrant builders in Shanghai.

Relationship of folic acid, homcysteine with pancreatic cancer: A case-control study / 第二军医大学学报

Objective: To study the relationship of folic acid, homocysteine with pancreatic cancer. Methods: A case-control study was performed in which the case group was defined as patients with newly diagnosed pancreatic cancer and the control group were population-based healthy individuals. ELISA assay was used to determine the plasma levels of folic acid, homocysteine, vitamine B6 and vitamine B12 in all the subjects. The general information (such as demology data, smoking history, diet, etc) was collected by face to face talking using a standard questionnaire. Univariate analyses were performed using Chi-square tests for norminal variables and unpair t-tests for continuous variables. The variables with a P value no more than 0.25 in univariate analyses were selected as candidate variables for a multivariate logistic regression analysis. Results: Forty-two patients with pancreatic cancer and 42 healthy individuals were included in the present study. The results of univariate analyses showed that the plasma folic acid, homocysteine, vitamine B12 and vitamine B6 were not significantly different between the two groups (P>0.05); they were potentially associated with pancreatic cancer (P0.05). The consumptions of vegetables, fruits, white meat and milk in case group was significantly less in the pancreatic cancer group than in the control group (P<0.05). Multivariate analysis showed that the odds ratios (95% CI) of plasma folic acid, vitamin B6, and homocysteine for pancreatic cancer were 0.571 (0.383-0.851), 0.750 (0.557-1.011), and 1.514 (0.986-2.326), respectively. Conclusion: Increased plasma folic acid can decrease the risk of pancreatic cancer. Plasma vitamin B6 might be a protective factor and homocysteine might be a risk factor of pancreatic cancer.

Infectious bovine rhinotracheitis viral gG expression and gG-ELISA development / 生物工程学报

Taking the genome DNA of Infectious Bovine Rhinotracheitis Virus (IBRV) as the template, the gG gene was amplified with PCR and cloned into the T cloning vector pMD18-T. After being identified by restriction digestion and DNA sequencing, the insert was subcloned into the expression vector pGEX-KG. Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that this gene was expressed as both soluble form and inclusion body by the transformed E. coli BL21 strain (DE3). The fusion protein was purified and used as the coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay (ELISA). Comparison between this gG-ELISA and commercial IBRV gB-ELISA Kit (IDEXX) was made in the detection of 380 cow serum samples. The results demonstrated an agreement of 92%. By using this novel gG-ELISA, 1248 cow serum samples were tested and the average positive rate of IBRV antibodies for imported cows is 21.7%, while the positive rate ranged greatly from 0.0%-41.5% for Hubei local Chinese Black and White Dairy Cows.

Preparation antibodies against recombinant bovine IFN-gamma and development of sandwich ELISA for bovine IFN-gamma detection / 生物工程学报

This study was aimed to establish ELISA for recombinant bovine IFN-gamma (BovIFN-gamma) detection and provide a new method for diagnosis of pathogenic infection. The total RNA was isolated from peripheral blood leucocytes cultured with PHA mitogen stimulation. Then bovine IFN-gamma (BovIFN-gamma) gene cDNA was amplified by RT-PCR and cloned into pET28a to obtain the expression plasmid designated as pETBovIFN-gamma. The pETBovlFN-gamma was further transformed into competent E. coli BL21 cells and a 18kD His-tagged protein as expected was expressed after IPTG induction. By using purified recombinant BovIFN-gamma as antigen and lymphocyte-hybridoma technique, four hybridoma cell lines which stably secreted monoclonal antibodies against rBovIFN-gamma were generated, designated as A7, A10, G6, and G10. The immunoglobin subset was identified as IgG1 . Western-blotting analysis and ELISA demonstrated that the monoclonal antibodies secreted by all the four hybridoma cell lines could react specifically to the recombinant BovIFN-gamma, but not irrelative proteins such as Ag85B, ESAT-6-CFP-10 and GM-CSF, suggesting that the four hybridoma cell lines were rBovIFN-gamma specific monoclonal antibodies. A sandwich ELISA was established by using A10 secreted monoclonal antibody and rabbit polyclonal antibodies against BovIFN-gamma, HRP labeled goat anti-rabbit IgG. The results indicated that the sensitivity was 2ng/mL. This sandwich ELISA to detect BovIFN-gamma paved the way to develop a sensitive method for specific infection detection such as bovine tuberculosis diagnosis.

Construction and characterization of delta crp delta asd mutant host-vector balanced lethal system of Salmonella choleraesuis C500 strain / 生物工程学报

Salmonella choleraesuis C500 strain was an attenuated vaccine strain to prevent piglet paratyphoid, attenuated by chemical method. Although the vaccine has good immunogenicity, it remains some residual virulence. In order to develop a safer vaccine strain and exploit C500 as a live vaccine vector for mucosal immunization, delta crp delta asd double deletion mutant was constructed. Firstly, the recombination suicide vector with 320 bp-deleted crp (cAMP receptor protein) gene and sacB (sucrose-sensitive gene) gene was constructed and conjugated with C500. The unmarked crp deleted strain without resistance was selected by two-step method and crp deletion on the genome was determined by PCR. Then the asd (beta-aspartic semialdehyde dehydrogenase) gene was further deleted in the delta crp strain by the same method. Foreign DAP (diaminopimelic acid) must be supplied for delta crp delta asd mutant to grow. The phenotype, growth properties and virulence in mice of delta crp mutant were further characterized. In conclusion, the delta crp delta asd double-deletion mutant was successfully constructed. The delta crp delta asd mutant can be used as a live vector to express foreign genes and to develop potential oral multivalent vaccines.