We investigated the mechanism of human aspartyl beta-hydroxylase (HAAH) in early diagnosis of tumors. The encoding gene of HAAH was cloned from the hepatic carcinoma by RT-PCR and expressed as a fused protein in the prokaryotic vector pBV-IL1. The expressed HAAH was purified by Ni(2+)-NTA purification column and the purified protein was then used to immunize Balb/c mice. Three hybridoma cell lines (respectively designated H3/E10, E4/F12 and G4/D8) stably expressing the monoclonal antibody specific to HAAH fusion protein were obtained. The specificity and sensitivity of the monoclonal antibody were assessed by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Finally, the monoclonal antibody expressed by H3/E10 cell line was used to detect the expression of HAAH in several tumor cell lines by indirect immuno-fluorescence, and the specific fluorescence was observed. In conclusion, this study successfully constructed the recombinant prokaryotic vector pBV-IL1-HAAH and prepared HAAH-specific monoclonal antibody for further study of the structure and function of the protein. The result may also lay solid foundation for the research of the molecular mechanism of HAAH in early diagnosis of tumors.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; genetics ; Humans ; Hybridomas ; metabolism ; Immunization ; Liver Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mixed Function Oxygenases ; biosynthesis ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

Objective To investigate the influence of blood glucose fluctuation on ser202 phosphorylation sites of tau protein( p-Tau) in hippocampus of diabetic rats; to explore the possible mechanism of blood glucose fluctuation impacting on tau protein hyperphosphorylation. Methods Healthy male Sprague Dawley rats were randomly divided into normal control group ( NC group ) and diabetes group. After diabetic rats model was established, all the diabetic rats were randomly divided into diabetic continuous hyperglycemia group (DC group) and diabetic blood glucose fluctuant group ( DF group). Rats in DF group were given glucose solution intraperitoneal injection twice at regular time everyday. 30 minutes after each intraperitoneal injection, insulin subcutaneously injections were given. Rats in the NC group and DC groups were given the same volume of saline subcutaneous injection. Specimens were collected in 8 weeks, the levels of p-Tau and total tau in rat hippocampus were detected by immunohistochemical staining and Western blotting. The immunoreactive positive products were analyzed by image analysis system. Glycogen synthase kinase-3β(GSK-3β) mRNA was detected by realtime PCR. Results (1) Blood glucose fluctuation of rats in DC and DF group were greater than NC group. And the mean blood glucose, standard deviation of mean blood glucose (SDBG), and large amplitude of glycemic excursion (LAGE) levels were increased significantly compared to NC group, the difference has statistical significance ( all P < 0. 05). Compared with DC group, SDBG and LAGE levels of DF group were higher (both P<0. 05). HbA1C and insulin levels were no difference (P>0. 05). (2) Compared with NC group, the hippocampal p-Tau level of DC group and DF group were increased (P < 0. 05 ); Compared with DC group, the hippocampal p-Tau expression of DF group was increased ( P <0. 05). Compared with DC group, a higher hippocampal GSK-3β mRNA level was found in DF group ( P <0. 05). Conclusions On the basis of diabetes animal model, giving glucose solution intraperitoneal injection and insulin subcutaneously injection 30 minutes later twice at regular time everyday could establish experimental model of diabetic blood glucose fluctuation. Blood glucose fluctuation may aggravate the diabetic rats hippocampal p-Tau. The possible mechanism seems to be an up regulation of the GSK-3β.

With completing a baseline survey of a large natural population cohort, conducting regular follow-up has become a key factor in further improving the quality of cohort construction and ensuring its sustainable development. Typical cohort follow-up methods include repeat surveys, routine monitoring, and community-oriented surveillance. However, in practical applications, there are often issues such as high costs, difficulty, and high error rates. Telephone follow-up is an important supplementary method to the methods mentioned above, as it has the characteristics of low cost, fast response, and high quality. However, the with difficult organization, quality control is challenging, response rates are low, and management levels vary widely, which limits its widespread use in large-scale population cohort studies. Given the above problems, this study draws on customer relationship management based on the actual needs of the China Northwest Cohort follow-up. It relies on the REDCap electronic data collection platform to build a telephone follow-up management and quality control system. Targeted solutions are provided for key issues in telephone follow-up implementation, including organizational structure, project management, data collection, and process quality control, to improve the quality control level of telephone follow-up comprehensively and thereby enhance the quality and efficiency of follow-up. We hope to provide standardized follow-up programs and efficient quality control tools for newly established and existing cohort studies.

Objective:To explore the disturbance of metal element balance in mice after exposure to radon.Methods:Mice were randomly divided into control group, radon exposure of 30 WLM group, 60 WLM group and 120 WLM groups, with 10 mice in each group. After radon exposure with the cumulative dose, the lung function of mice was detected by a non-invasive pulmonary function testing instrument. Mice blood was taken from eyeballs. The lungs, heart, liver, kidney and spleen were also collected. HE staining was used to observe the pathological changes of lung tissue. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect the content of metal elements, including essential trace elements in the body: chromium (Cr), molybdenum (Mo), cobalt (Co), selenium (Se), copper (Cu), zinc (Zn), manganese (Mn), nickel (Ni), and potentially toxic elements: arsenic (As), tin (Sn), lead (Pb), aluminum (Al), mercury (Hg), cadmium (Cd), and silver (Ag).Results:Compared with the control group, lung ventilation function of the radon-exposed mice was decreased, alveolar structure was destroyed, and the contents of pulmonary metal elements Cr, Al, Pb, Sn( F=0.34, 0.66, 3.14, 1.16, P<0.05) and essential trace elements Mn, Cr, Zn, and Mo in the blood were decreased( F=0.65, 1.44, 0.97, 2.08, P<0.05), while the elements of Cu, Mo, Se and As in the lungs were increased( F=1.31, 1.26, 0.81, 2.04, P<0.05), and the element contents in other tissues also fluctuated. Conclusions:Inhalation of a certain cumulative dose of radon can reduce the lung ventilation function of mice and induce lung inflammation, as well reduce the content of essential trace elements in the lung and blood so that the content of metal elements in the body fluctuates.

Pulmonary fibrosis(PF)is a progressive,interstitial fibrotic lung disease characterized by persistent scar formation in the lung parenchyma,and a reduced quality of life and poor prognosis for patients.The pathogenesis of PF is unknown and there is a lack of effective therapeutic agents;however,animal models are currently the main tool used to explore the pathogenesis of the disease and to find effective therapeutic agents.PF can be induced by various factors and to different degrees according to known etiologies.Among these,bleomycin-induced models are widely used because of their reproducibility and the similarity between the fibrosis pathology and clinical conditions.The main induction method include intratracheal drip,intratracheal nebulization,tail vein injection,intraperitoneal injection,and transnasal inhalation,and these can be classified into single and multiple doses,according to the frequency of induction.Based on the relevant literature,the current review summarizes the characteristics of the bleomycin-induced PF model using different induction frequencies and method,to provide a basis for the application of this model.

Cohort study is one of the important research methods in analytical epidemiology because of its clear time sequence relationship, which is better than other observational studies in demonstrating causal association. However, screening diagnosis or other methods are often used to exclude the individuals with outcome events during the enrollment process of the subjects in cohort studies. The accuracy of screening diagnosis and the effectiveness of exclusion will affect the accuracy of the baseline status assessment of the subjects included in the study, which may lead to the causal time sequence reversal of exposure-outcome in the estimation of causal effect. Landmark analysis can be used to control reverse causality by excluding subjects with potentially unknown expose-outcome timing. In this paper, we describe the basic principles and analytical steps of landmark analysis, and use data from the Chinese Longitudinal Healthy Longevity Survey to explore the relationship between physical activity and frailty, and introduce the specific application of landmark analysis for the purpose of facilitating its application and inferring causal effects more accurately in cohort studies.