Abstract: Objective To investigate the expression of T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) on the surface of T cells in patients with brucellosis (Bm), as well as the expression of interleukin-10 (IL-10) and transforming growth factor beta (TGF-β) in serum, and to analyze the differential expression of these indicators in patients with acute and chronic brucellosis, in order to provide new approaches for the differential diagnosis of acute and chronic brucellosis. Methods A total of 56 patients diagnosed with brucellosis at the First Affiliated Hospital of Xinjiang Medical University from April 2023 to September 2023 were selected, including 31 patients in the acute phase and 25 patients in the chronic phase. Additionally, 35 healthy individuals underwent routine physical examinations within the same period served as healthy controls. Flow cytometry was used to detect and compare Tim-3 levels on the CD4+ T cells' surface among the groups. Levels of serum IL-10 and TGF-β were measured and compared using CBA and ELISA, respectively, and the relationship of these factors with the staging of brucellosis patients was analyzed. Results The proportions of Tim-3+CD3+CD4+T cells in the control group, acute group, and chronic group were (2.56±1.25)%, (5.14±1.98)%, and (13.66±2.66)%, respectively. The Tim-3 levels in the patients with brucellosis were higher than those in the healthy control group, with the chronic group showing even higher levels, and these differences were statistically significant (P<0.05). The levels of IL-10 and TGF in the patient group were higher than those in the healthy control group, with the chronic group exhibiting significantly higher levels of IL-10 and TGF-β than the acute group, also presenting statistically significant differences (P<0.05). The areas under the ROC curve for predicting chronic brucellosis with Tim-3, IL-10, and TGF-β scores were 0.876, 0.865, and 0.663, respectively. Conclusions There are certain differences in the expression of Tim-3, serum IL-10, and TGF-β among patients with brucellosis, with high expression indicating a potential transition to the chronic phase of the disease. Tim-3 has shown the best diagnostic performance. Therefore, as a diagnostic indicator, Tim-3 may provide new ideas and strategies for the treatment and differential diagnosis of brucellosis.

Objective To dynamically analyze the antibodies with regard to in vitro serum bacteri-cidal activity and the quantity of IgG in healthy adults received one dose of immunization with ACYW 135 me-ningococcal polysaccharide vaccine .To investigate the term of protection with polysaccharide vaccine and the correlation between bactericidal titer and IgG concentration .Methods Twenty healthy adults were given one dose of immunization with quadrivalent meningococcal polysaccharide vaccine .Serum samples were col-lected before and after vaccination at specific time points .Test for serum bactericidal activity ( SBA ) and quantitative ELISA were performed to detect bactericidal titers and IgG concentrations in serum samples .Re-sults Certain levels of antibodies against capsular polysaccharides of serogroup A , C, Y and W135 strains had already existed prior to immunization .Moreover, bactericidal titers against serogroup A , Y and W135 strains except serogroup C strain were relatively high .Specific IgG concentrations and bactericidal titers for all serogroup stains were significantly increased on day 15 after vaccination (P<0.05), and then sustained at a high level during a time period from day 30 to day 160.Both IgG concentrations and bactericidal titers dropped to the levels similar to that before preimmunization in about 3 years.No significant correlation was observed between bactericidal titers and IgG concentrations (r<0.3, P>0.05).However, a close correla-tion was demonstrated between GMTs and GMCs of serum samples (r>0.7, P<0.05).Conclusion The geometric mean titers ( GMTs) and geometric mean concentrations ( GMCs) of serum samples collected be-fore and after vaccination at different time points were reliable and consistent parameters for the evaluation of vaccine .The term of protection of quadivalent meningococcal polysaccharide vaccine was about 3 years upon a single dose of immunization .

Objective To re-evaluate the immunogenicity of meningococcal serogroups A and C polysaccharide vaccine among a healthy population of age 5 to 59.Methods Pre and post-vaccination ser-um samples were collected from the subjects involved in a randomized , controlled clinical trial conducted in 2005 at Hechi, Guangxi, China.Enzyme-linked immunosorbent assay (ELISA) was performed for the quan-titative detection of specific anti-capsule IgG antibody against meningococcal serogroups A and C in serum samples.Serum bactericidal assay ( SBA) was used to measure the bactericidal antibody activity against se-rougroups A and C bacterial strains .Results Geometric mean concentrations (GMCs) of specific anti-cap-sule IgG antibody were 23.66 μg/ml and 78.83 μg/ml for serogroup A (P<0.05), and 1.23 μg/ml and 51.25 μg/ml for serogroup C (P<0.05) in serum samples of pre and post-vaccination, respectively.Be-fore immunization , 99% of serum samples ( 97/98 ) showed serogroup A-specific IgG concentration ≥2μg/ml, which reached up to 100%(98/98) after vaccination (P>0.05).The percentage of serum sam-ples with serogroup C-specific IgG concentration ≥2 μg/ml rose from 20%to 99% after vaccination ( P<0.05).The ratio of positive serum samples with at least four times of increases in concentration of specific IgG against serogroup A and serogroup C after vaccine immunization were 34% and 100%, respectively . The rSBA geometric mean titers ( GMTs) for serogroups A and C in serum sample of post-vaccination were respectively elevated from 1164 to 5530 (P<0.05), and from 4 to 6225 (P<0.05).The percentage of se-rum samples with rSBA GMTs≥128 increased from 91% to 99% for serogroup A (P>0.05), and from 14%to 96%for serogroup C (P<0.05) after vaccination.The rSBA GMTs with at least four times of in-creases after vaccination were detected respectively in 53% and 100% of serogroups A and C vaccinated subjects.Pearson correlation coefficient between IgG concentration and rSBA GMTs was r=0.15 (pre) and r=0.23 (post) for serogroup A, and r=-0.14 (pre) and r=0.58 (post) for serogroup C (P<0.05). Conclusion This study has demonstrated an efficient and sustained immunogenicity with meningococcal sero -groups A and C polysaccharide vaccine as evidenced by the data from standardized assays of ELISA and SBA .

Objective To monitor and analyze the antigenicity of Streptococcus pneumonia polysac-charide, its derivatives and conjugates by three immunological assays .Methods Inhibition ELISA and rate nephelometry(RN) were established for this study.Antigenicity of serotype 23F pneumococcal conjugates and their intermediates were analyzed by double immunodiffusion assay , inhibition ELISA and RN .The re-sults derived from three assays were comparatively analyzed to evaluate the changes of antigenicity during the preparing process of serotype 23F conjugate.Results Double immunodiffusion assay, inhibition ELISA and RN were all applicable to antigenicity analysis during the process of conjugate preparation .Inhibition ELISA could quantitatively detect a slight difference of polysaccharide antigenicity during the preparing process . Conclusion The antigenicity of polysaccharide during the preparing process of pneumococcal conjugates could be analytically monitored by using three immunological assays .This study provided evidence for suc-cessfully using immunological assays as the quality control means during the preparing process of pneumococ -cal conjugates .

Objective To establish a standardized quantitative enzyme-linked immunosorbent as-say ( ELISA) recommended by WHO for the detection of human IgG antibodies specific for Streptococcus pneumoniae capsular polysaccharides ( Pn PS ELISA ) .Methods According to the WHO recommended standard Pn PS ELISA protocol , capsular polysaccharide concentrations of 13 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F) of Streptococcus pneumonia for coating were optimized;the ELISA plates and AP conjugated goat anti-human IgG antibody for the detection were experimentally selected .Using the established assay parameters assured by testing quality control standards ( QCs) provided by WHO refer-ence laboratory , a panel of 16 LIBP ( Lanzhou Institute of Biological Products Co .Ltd.) QCs were measured for comparison analysis between WHO reference laboratory and LIBP laboratory .In the meantime , the range of IgG concentrations of an internal QC panel 907 for 13 serotypes of pneumococcal capsular polysaccharide were established for routine QC work in LIBP laboratory , and inter-assay precision within LIBP laboratory was assessed as well .Results The standardized Pn PS ELISA assay established in LIBP laboratory met the criteria required by WHO .There was a high correlation between the data collected by WHO reference labora -tory and LIBP laboratory (slope=0.94, coefficient of correlation r=0.97, P<0.05).Eighty one percent of IgG concentrations measured by LIBP laboratory were within the range of ±40%of those measured by WHO reference laboratory , which met the criteria of 75%data falling within the ambit of ±40%of assigned values commonly used for comparison .The ranges of IgG concentration in LIBP QC 907 for 13 serotypes had been established.The inter-assay precision in LIBP laboratory was high with coefficiency of variation ( CV) less than 30%.Conclusion LIBP laboratory has successfully established the standardized ELISA recommended by WHO for quantitative detection of human IgG antibodies against pneumococcal capsular polysaccharides (Pn PS ELISA).The range of IgG concentrations in LIBP QC 907 for 13 serotypes of pneumococcal capsular polysaccharide are also established for routine quality-control practice .

Objective To prepare a human meningococcal reference serum and standardize IgG concentrations to capsular polysaccharides and in vitro bactericidal activities of the reference serum against serogroup A, C, Y and W135 strains.Methods Twenty healthy adults were recruited and given one dose of immunization with tetravalent (serogroups A, C, Y and W135) meningococcal polysaccharide vaccine . Plasma samples were collected and gone through a series of process treatments including defibrination , filtra-tion, and lyophilization to prepare the meningococcal reference serum Men 10.The IgG concentrations of Men10 to capsular polysaccharides of serogroups A , C, Y and W135 were calibrated by using an internation-al reference serum CDC1992 as the standard in enzyme-linked immunosorbent assay (ELISA).Provisional IgG concentrations of Men10 were intensively validated by testing a panel of 12 calibration serum samples from Centers for Disease Control and Prevention , USA ( US CDC) and a panel of 56 serum samples immu-nized with A, C, Y and W135 meningococcal polysaccharide vaccine from Lanzhou Institute of Biological Products Co., Ltd.(LIBP) with the assays using Men10 and CDC1992 as the standard and/or test sam-ples, respectively.The bactericidal titers against serogroup A , C, Y and W135 strains were measured by se-rum bactericidal assay (SBA).Results Four thousand vials (0.5 ml/vial)of lyophilized human meningo-coccal reference serum Men10 were successfully prepared with 2.5%of residual moisture .Reference serum Men10 was sterile and free from contamination by hepatitis B virus , hepatitis C virus , human immunodefi-ciency virus and syphilis .Provisional IgG concentration of Men 10 to capsular polysaccharide of serogroups A, C, Y and W135 was calibrated by using CDC1992 as the standard.Furthermore, IgG concentrations of both panels of 12 CDC calibration serum samples and 56 LIBP serum samples calibrated by using Men 10 as the standard correlated well with those by using CDC1992 as the standard (r=0.99,P<0.05).The IgG concentrations of CDC1992 as calibrated by using Men10 as the standard showed significant correlation with its previously determined values with variation <10%.SBA titers for serotype A , C, Y and W135 strains were established as well .Conclusion A panel of new human meningococcal reference serum Men 10 with accurately calibrated IgG concentration against capsular polysaccharide of serogroups A , C, Y and W135 as well as SBA titers was successfully established .

Objective To observe the effects of salidroside regulating glucose metabolism in type 2 diabetic mice,then to explore the molecular mechanism. Methods Type 2 diabetes model was induced by feeding high-fat diet and intraperitoneal injecting STZ to male C56BL/6J mice,then the glucose related indexes,micro RNA-370 levels in the serum and liver tissue and the expression of gluconeogenesis key protein(G6Pase and PEPCK) in the liver tissue to observe the treatment effects of salidro-side on type 2 diabetic-caused gluose metabolic disorder.In cell test,we isolated primary hepatocytes,then silenced or over-ex-pressed micro RNA-370 in mouse primary hepatocytes to observe the molecular mechanism of glucose metabolic regulation of the micro RNA-370 and salidroside. Results Treated with salidroside 40,80 and 160 mg·kg-1,the results showed that compared with the model control group,the glucose related indexes were all improved significantly.The relative expression levels of micro RNA-370 in serum and liver,and that of PEPCK and G6Pase all reduced in different degrees,dose-dependently.The changes of middle and high dose group decreased significantly(P<0.05),that of low dose group had a decreasing trend but no statistically significant.In the cell test,compared with the normal control group,salidroside alone group and micro RNA-370 inhibitor group were able to reduce the protein expression level of PEPCK and G6Pase(P<0.05),micro RNA-370mimic alone group can signifi-cantly increase the protein expression level of PEPCK and G6Pase(P<0.05),compared with the micro RNA-370mimic alone group,combining micro RNA-370 mimic and salidroside can significantly reverse the increasing caused by micro RNA-370 mimic alone(P<0.05). Conclusion Our research found that salidroside can improve glucose metabolism disorder in type 2 diabetic mice,and at least in part,through the suppression of micro RNA-370 expression for the first time.

Objective To compare the clinical characteristics and influence factors of hyperuricemia between genders,in order to provide references for better controlling and preventing the occurrence and development of hyperuricemiaprovide.Methods A total of 5 783 people who underwent physical examination in two Baoding Health Screening Centers from January 1st to June 1st,2016 were enrolled in this study,and all volunteers completed physical examination,laboratory examination and questionnaire survey.Patients with hyperuricemia were selected to analyse the clinical characteristics and influence factors.Results There were statistically significant differences in clinical characteristics,including obesity,blood pressure,blood glucose,blood lipids,liver function,renal function,anemia,blood rheology examination,thyroid ultrasound and lateral radiographs,between male and female patients with hyperuricemia (P＜0.05).The survey showed that there were statistically significant differences in age,education level,marital status,work status,sleeping status,smoking and drinking between male and female patients with hyperuricemia (P＜0.05).The red blood cells counts,marital status and education level were influence factors for female patients with hyperuricemia,while have little effect on male patients.The smoking,creatinine and diastolic blood pressure were influence factors for male patients with hyperuricemia,while have no effect on female patients.Conclusion The clinical characteristics and the influencing factors of male and female patients with hyperuricemia are different,so corresponding preventive and therapeutic measures should be taken for male and female patients.

Objective:To analyze the correlation between two-dimensional ultrasound and HbA1c, Thyroid stimulating hormone (TSH) and its diagnostic value in benign and malignant Thyroid nodules.Methods:A total of 155 patients with thyroid nodules examined in the Department of Ultrasound Medicine of Haikou Orthopedics and Diabetes Hospital from January 2020 to January 2021 were prospectively selected. With surgical pathological examination as the gold standard, they were divided into benign group (75 cases) and malignant group (80 cases) according to the nature of nodules. There were 40 males and 35 females in the benign group, with an age of (47.95±8.26) years old and BMI of (22.45±1.26) kg/m2. There were 42 males and 38 females in the malignant group, with an age of (48.22±8.01) years old and BMI of (22.36±1.21) kg/m2. The expression level of HbA1c was detected by automatic biochemical analyzer, and the concentration of TSH was detected by automatic immunoassay. SPSS 19.0 statistical software was used for analysis and processing.Results:Compared with benign thyroid nodules, the mean transit time (MTT) and time to peak intensity of malignant thyroid nodules were (0.92±0.11 vs 1.48±0.42) s and (1.48±0.42) s respectively. TIP (1.06±0.21 vs 1.57±0.56) s, perfusion Index (PI) (1.15±0.22 vs 1.76±0.62) s parameters increased, and the differences were statistically significant ( P < 0.05) . Compared with patients with benign thyroid nodules, the expression levels of HbA1c (6.98±0.73 VS 10.22±1.35) % and TSH (3.95±0.53 VS 8.75±0.95) mIU/L in patients with malignant thyroid nodules were significantly higher ( P<0.05) . The expression levels of MTT, TIP, PI, HbA1c, and TSH were correlated with those of benign and malignant thyroid nodules, and the expression levels of MTT, TIP, PI, HbA1c, and TSH were positively correlated with those of benign and malignant patients ( P<0.05) . MTT, TIP and PI had correlation with the expression levels of HbA1c and TSH. Positive correlation: MTT had a positive correlation with HbA1c and TSH. TIP had a positive correlation with HbA1c and TSH. PI had a positive correlation with HbA1c and TSH ( P<0.05) . Compared with the single diagnosis of MTT, TIP and PI, the three combinations had higher diagnostic value for malignant thyroid nodules ( P=0.001) . Conclusion:Two-dimensional ultrasound parameters have high diagnostic value in malignant thyroid nodules, and have certain correlation with HbA1c and TSH, which can be used for the diagnosis of malignant thyroid nodules.

Objective To summarize and evaluate the characteristics of current recurrent spontaneous abortion(RSA)animal models at home and abroad,and to provide reference and guidance for the standardized preparation of RSA models.Methods"Recurrent spontaneous abortion"and"animal model"were used as co-keywords in CNKI,Wanfang,VIP,PubMed and Web of Science databases to search the RSA animal experimental literature,covering the period up to January 20,2024,and a total of 1 411 articles were collected.The analysis focused on construction methods and essential elements of RSA animal models,the modeling process and result evaluation,as well as the application of these models in pharmacological and pharmacodynamic research.An Excel table was established for systematic analysis and discussion.Results A total of 138 experimental studies were obtained after screening.In constructing RSA animal models,immunological models were the most widely used in Western medicine(96.92％),with the Clark model being the main one(92.31％).In traditional Chinese medicine(TCM)models,70.00％were kidney deficiency-luteal inhibition-syndrome combination models,20.00％were kidney deficiency and blood stasis models,and 10.00％were deficiency-heat syndrome models.Most animals were selected at 6-8 weeks(33.86％)and 8 weeks(32.28％)of age.The majority of animals were paired for mating at 18:00 on the day of cage pairing.In 81.03％of literatures,vaginal plugs were checked once the following morning,with 8:00 being the most common time(17.02％).The most commonly used drug administration cycle was 14 days of continuous gavage after pregnancy.Among the tested drugs,Western drugs were mainly protein-based(29.17％),while TCM drugs were mainly TCM decoction(81.11％).The most frequently used methods for detecting indicators included visual observation of embryos(22.54％),western blot(15.96％),PCR(13.58％),ELISA(12.91％),HE staining(10.80％)and immunohistochemistry(9.39％).Conclusion The etiology of RSA is complex,and corresponding animal models should be established based on different etiologies.Clark model is commonly used in the construction of Western medicine model,while the kidney deficiency-luteal inhibition-syndrome combination model is predominant in TCM.RSA animal model is widely used in related research,but systematic evaluation needs to be strengthened.