Objective To analyze the level of 25-hydroxyvitamin D[25(OH)D]in patients with type 2 diabetes mellitus,and preliminarily investigate the correlation between serum 25(OH)D level and metabolic indicators such as glycosylated hemoglobin(HbA1c)and pancreatic islet function in patients with type 2 diabetes mellitus.Methods A total of 459 patients with type 2 diabetes mellitus who were hospitalized in the Department of Endocrinology,Xinxiang First People's Hospital from January 2020 to December 2020 were selected as the research subjects.Clinical data of the patients were collected,including gender,age,serum 25(OH)D,fasting insulin,C-peptide,HbA1c,fasting blood glucose,postprandial blood glucose,urinary microalbumin,urinary albumin-to-creatinine ratio,blood calcium,blood uric acid(UA),triglyceride(TG),total cholesterol(TCH),low-density lipoprotein(LDL),and high-density lipoprotein(HDL).According to the serum 25(OH)D level,the patients were divided into sufficient group[n=20,25(OH)D≥30 μg·L-1],insufficient group[n=95,20 μg·L-125(OH)D＜30 μg·L-1],deficient group[n=231,10 μg·L-1 ≤25(OH)D＜20 μg·L-1],and severely deficient group[n=113,25(OH)D＜10 μg·L-1].Differences in metabolic indicators of patients in the four groups were compared,and the correlation between 25(OH)D level and metabolic indicators was analyzed by using the Pearson correlation.Results The serum 25(OH)D level of patients with type 2 diabetes mellitus was 3.00-46.59(15.75±0.35)μg·L-1;the serum 25(OH)D level of male patients was significantly higher than that of female patients(P＜0.05).The prevalence of 25(OH)D deficiency in patients with type 2 diabetes mellitus was 74.9％(344/459).The 25(OH)D deficiency mainly occurred in January,February,March,April,November,and December.Patients in the insufficient,deficient,and severely deficient groups had significantly higher HbA1c levels than those in the sufficient group(P＜0.05),and the HbA1c levels of patients in the deficient and severely deficient groups were significantly higher than those in the insufficient group(P＜0.05);there was no statistically significant difference in the HbA1c level between the deficient group and the severely deficient group(P＞0.05).There was no statistically significant difference in fasting blood glucose between the sufficient group and insufficient group,and between the deficient group and severely deficient group(P＞0.05);fasting blood glucose of patients in the deficient and severely deficient groups was significantly higher than that in the sufficient and insufficient groups(P＜0.05).There was no statistically significant difference in fasting insulin,urinary microalbumin,daily total urinary albumin,and urinary albumin-to-creatinine ratio of patients among the sufficient,insufficient,and deficient groups(P＞0.05);the fasting insulin of patients in the severely deficient group was significantly lower than that in the sufficient,insufficient,and deficient groups(P＜0.05);the urinary microalbumin,daily total urinary albumin,and urinary albumin-to-creatinine ratio of patients in the severely deficient group were significantly higher than those in the sufficient,insufficient,and deficient groups(P＜0.05).There was no statistically significant difference in the homeostasis model assessment for insulin resistance(HOMA-IR),serum albumin,blood creatinine,1-hour postprandial blood glucose,2-hour postprandial blood glucose,3-hour postprandial blood glucose,fasting C-peptide,1-hour postprandial C-peptide,2-hour postprandial C-peptide,3-hour postprandial C-peptide,TG,TCH,LDL,HDL,blood UA,and blood calcium of patients among the four groups(P＞0.05).Pearson correlation analysis showed that serum 25(OH)D levels in patients with type 2 diabetes mellitus were negatively correlated with HbA1c,urinary microalbumin,and urinary albumin-to-creatinine ratio(r=-0.093,-0.166,-0.157;P＜0.05),and positively correlated with fasting insulin(r=0.089,P＜0.05).Serum 25(OH)D levels in patients with type 2 diabetes mellitus had no correlation with fasting blood glucose,HOMA-IR,serum albumin,blood creatinine,1-hour postprandial blood glucose,2-hour postprandial blood glucose,3-hour postprandial blood glucose,fasting C-peptide,1-hour postprandial C-peptide,2-hour postprandial C-peptide,3-hour postprandial C-peptide,TG,TCH,LDL,HDL,blood UA,and blood calcium(P＞0.05).Conclusion 25(OH)D deficiency and insufficiency are common in patients with type 2 diabetes mellitus,especially in female patients.In patients with type 2 diabetes mellitus,25(OH)D level is positively correlated with fasting insulin and negatively correlated with HbAlc,urinary microalbumin,and urinary albumin-to-creatinine ratio.25(OH)D deficiency in patients with type 2 diabetes mellitus is mainly distributed in January,February,March,April,November,and December.

We investigated the enhanced immune response of a recombinant T cell immunogen as an effective cellular immune adjuvant. The T cell immunogen named TI contained several T cell epitopes from the VP1, VP4, 3A and 3D proteins of foot-and-mouth disease virus (FMDV) and two pan-T helper (T(H)) cell sites to broaden the immunogenicity of the protein. Meanwhile, another fusion protein named OA-VP1 was expressed in bacteria, which contained two VP1 proteins of O and Asia1 type FMDV. Mice were vaccinated with commercially inactivated vaccine or OA-VP1 protein with or without the TI immunogen. The results show that mice inoculated with inactivated vaccine or OA-VP1 protein supplemented with TI immunogen produced significantly higher level of neutralizing antibodies (P < 0.01 or P < 0.05) than the mice only inoculated with inactivated vaccine or OA-VP1 protein by microneutralization assay. An obvious increase in T cell number by flow cytometric analysis and significantly higher concentration of IFN-gamma secreted in culture media of spleen lymphocytes were observed in groups supplemented with TI immunogen (P < 0.01). TI immunogen was an effective stimulator for humoral and cellular immunity and could help improve the immunogenicity of inactivated vaccine or protein subunit vaccine.
Adjuvants, Immunologic ; pharmacology ; Animals ; Capsid Proteins ; genetics ; immunology ; Epitopes, T-Lymphocyte ; genetics ; immunology ; Foot-and-Mouth Disease ; immunology ; prevention & control ; virology ; Foot-and-Mouth Disease Virus ; immunology ; Immunization ; Mice ; Viral Vaccines ; genetics ; immunology ; pharmacology

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Foot-and-Mouth Disease/prevention & control* ; Foot-and-Mouth Disease Virus ; Viral Nonstructural Proteins/genetics* ; Viral Vaccines