convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Objective:To study the effect of Compound Duzhong Jiangu Granules on joint function, quality of life and inflammatory factors in patients with Kashin-Beck disease.Methods:By group design, 135 patients with Kashin-Beck disease in Chongxin County, Pingliang City, Gansu Province, were selected and divided into intervention group (100 cases treated with Compound Duzhong Jiangu Granules, 12 g/bag, 1 bag/time, 3 times/day, treatment for 1 month) and control group (35 cases treated with ibuprofen, 0.3 g/capsule, 1 capsule/time, 2 times/day, treatment for 2 weeks) according to the randomized, single-blind principle. The changes of joint dysfunction index score, joint function improvement rate, total scores of Kashin-Beck disease quality of life (KBDQOL) and scores of each dimension were analyzed before treatment, 1 month and 3 months after treatment. The serum levels of interleukin-6 (IL-6), nuclear factor κB (NFκB)-p65, inducible nitric oxide synthase (NOS2), nitric oxide (NO), and prostaglandin E2 (PGE2) were detected by enzyme-linked immunosorbent assay before treatment and 1 month after treatment.Results:The total scores of joint dysfunction index of the intervention group and the control group 1 month after treatment and 3 months after treatment were lower than those before treatment, but 3 months after treatment was higher than 1 month after treatment ( P < 0.001). One month after treatment, the total effective rates of joint function improvement in the intervention group and the control group were 68.00% (68/100) and 54.55% (18/33), respectively; 3 months after treatment, the total effective rates of the intervention group and the control group were 36.00% (36/100) and 39.39%(13/33), respectively. The total scores and scores of each dimension of KBDQOL were not significant for the main effect of the group ( P > 0.05), but significant for the main effect at the time point ( P < 0.05), and there was no interactive effect ( P > 0.05). There were significant differences in the scores of social support and mental status dimensions in the intervention group at different time points ( P < 0.001). And in the intervention group, the total score, the scores of physical function, activity limitation, economy and overall health dimensions were statistically significant between before treatment and 1 month after treatment, 1 month after treatment and 3 months after treatment( P < 0.05); however, there was no significant difference between before treatment and 3 months after treatment ( P > 0.05). One month after treatment, the serum PGE2 levels of both groups were decreased ( P < 0.05), and there was no difference in other inflammatory factors at different groups and time points ( P > 0.05). Conclusion:Compound Duzhong Jiangu Granules can effectively inhibit the inflammatory reaction of patients with Kashin-Beck disease, promote the improvement of joint function and improve the quality of life in various aspects.

Objective:To explore the ability of sequential immunization regimen inducing neutralizing antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) Wuhan-Hu1 and Omicron variants in mice.Methods:Groups of 6-8-week-old BALB/c mice were primed with two doses of Wuhan-Hu-1 inactivated vaccine, and then boosted with Omicron or Wuhan-Hu-1 inactivated vaccine, respectively. Binding antibodies were tested by enzyme linked immunosorbent assay; and neutralizing antibodies against Wuhan-Hu1 and Omicron variants were analyzed by vesicular stomatitis virus pseudovirus assay system; SARS-CoV-2 specific cellular immune responses were quantified by enzyme-linked immunosorbent spot assay.Results:IgG antibodies against Wuhan-Hu1, Delta and Omicron RBD were enhanced after the second dose of Wuhan-Hu1 inactivated vaccine. Compared with Wuhan-Hu1 inactivated vaccine, the group boosted with Omicron inactivated vaccine improved Wu-RBD and Omic-RBD specific IgG antibodies 1.41 and 1.26 times, and serum neutralizing antibodies against BA.1, BA.2, BA.4/5 and BF.7 were elevated 4.5, 3.4, 12.1 and 6.5 folds, respectively, by sequential immunization. After booster immunization with inactivated Wuhan-Hu1 or Omicron vaccines, Wu-RBD IgA titer was significantly higher than that of one dose inactivated Wuhan-Hu1 vaccine ( P=0.005 7, P=0.006 1). Conclusions:Neutralizing antibodies against Omicron variants were enhanced by sequential immunization with Omicron inactivated vaccine. Specific IgA was significantly enhanced after the third dose of inactivated vaccine.

Objective:To investigate the effects and potential mechanism of vitamin D supplementation on testicular function in aging rats induced by D-galactose.Methods:The aging rats were induced by D-galactose with subcutaneous injection. The animals were randomly divided into 6 groups: aging rats (DG), aging rats with low-dose vitamin D supplementation (LD), aging rats with high-dose vitamin D supplementation (HD), normal control rats(NC), normal rats with low-dose vitamin D supplementation(LN), normal rats with high-dose vitamin D supplementation (HN). The body weight, testicular weight, serum testosterone concentrations and sperm quality of the rats in each group were measured. The testis morphological changes were detected using light microscopy. The activity of superoxide dismutase (SOD) and level of malondialdehyde(MDA) were determined with spectrophotometer. The expression levels of Nrf2, GCLC, SOD2 and VDR in testis were detected by western blot.Results:At baseline, compared with NC group, testicular weight, serum testosterone level, SOD activity, Nrf2, GCLC and SOD2 expression levels were significantly decreased in DG group, while MDA level was significantly increased. After vitamin D supplementation, testicular weight, testosterone levels and SOD activity in both of HD and LD groups were significantly increased, while the MDA level was significantly decreased. The expression levels of Nrf2, GCLC, SOD2 and VDR were significantly increased.Conclusion:Vitamin D supplementation may enhance the testicular antioxidant capacity through activating Nrf2-ARE signaling pathway, and improve the testicular function in D-galactose-induced aging rats.

Objective:To evaluate the reliability and validity of the Chinese version of concise health status questionnaire (SF-36 scale) in evaluating the quality of life of patients with chronic Keshan disease, and to provide a scientific basis for studying the quality of life and the evaluation of treatment and rehabilitation of this population.Methods:In the August 2017, using cluster random sampling method, 175 patients with chronic Keshan disease treated by self-management of family beds in Pingliang City, Gansu Province in 2017 were selected as survey subjects, and demographic and disease data were collected. The Chinese version of SF-36 scale was used to investigate the quality of life. Split-half reliability and Cronbach's α coefficient were used to evaluate the reliability of the SF-36 scale; the factor analysis, correlation and differences between groups were used to evaluate the validity of the SF-36 scale.Results:The split-half reliability value of SF-36 scale was 0.916, and the Cronbach's α coefficient was 0.869. Factor analysis extracted 3 common factors from 8 dimensions of SF-36 scale, and the cumulative contribution rate of the 3 common factors to the total variance was 72.08%. In addition to the correlation coefficient ( r) between Role-Emotional and Bodily Pain dimension, the r value between total score and the scores of each dimension, and the scores of each dimension of SF-36 scale were 0.140 - 0.769. Except for the Bodily Pain dimension, there were statistically significant differences in the scores of Physiological Functioning, Role-Physical, General Health, Vitality, Social Functioning, Role-Emotional, and Mental Health dimension of the quality of life of patients with different grades of cardiac function ( F = 4.66, 10.73, 6.77, 14.61, 5.58, 9.57, 7.10, P < 0.05). Conclusion:The Chinese version of SF-36 scale has good reliability and validity in evaluating the quality of life of patients with chronic Keshan disease, and can be used to evaluate the quality of life of the patients.

Objective:To evaluate the reliability and validity of Chinese version of Minnesota Living with Heart Failure Questionnaire (MLHFQ) scale among patients with chronic Keshan disease.Methods:The patients with chronic Keshan disease diagnosed in Keshan disease area of Gansu Province from August 2018 to April 2020 were selected as the survey subjects to analyze the reliability and validity of the Chinese version of MLHFQ scale in evaluating the quality of life of patients with chronic Keshan disease. To evaluate the reliability of MLHFQ scale, the Cronbach's α coefficient was used, and when the Cronbach's α coefficient > 0.8 indicated that the consistency reliability of the scale was good. Kaider-Meyer-Olkin(KMO) statistic and Barlett's sphericity test were used to evaluate whether the scale was suitable for factor analysis (applicable when KMO > 0.6 and Barlett's sphericity test P < 0.05). The principal component analysis and maximum variance method of exploratory factor analysis were used to extract common factors, and the structural validity of the scale was evaluated. The discrimination validity of the scale was evaluated by New York Heart Association (NYHA) cardiac function classification and analysis of variance. Pearson correlation analysis was used to evaluate the content validity of the scale. Results:A total of 333 questionnaires were collected, of which 329 were valid. There were 180 males and 149 females, aged (59.52 ± 9.26) years. The Cronbach's α coefficient of the Chinese version of MLHFQ scale was 0.95 ( > 0.8). Among them, the Cronbach's α coefficients in the physical domain, emotional domain and other domains were 0.93, 0.91 and 0.86, respectively ( > 0.8). The KMO value of the Chinese version of MLHFQ scale was 0.95 ( > 0.6), and Barlett's sphericity test rejected the hypothesis (χ 2 = 5 222.01, P < 0.05). Principal component analysis and maximum variance method were used to extract 3 common factors, which were named kinetic energy of life, physical condition and emotional condition, respectively. The cumulative contribution rate of principal components was 66.22% ( > 50%). Compared with patients with NYHA cardiac function grade Ⅱ and patients with grade Ⅲ and Ⅳ, there were significant differences in the physical domain, other domain and total score ( P < 0.001). The Pearson correlation coefficients between the physical, emotional, and other domains and their respective items ranged from 0.74 to 0.88, 0.78 to 0.90, and 0.56 to 0.80, respectively, and the correlation coefficients between each item and its domain were all > 0.5 ( P < 0.001). Conclusion:The Chinese version of MLHFQ scale has good reliability and validity in patients with chronic Keshan disease in Gansu Province, and can be used to evaluate the quality of life of patients with chronic Keshan disease.

Objective:To evaluate the effect of self-management treatment for patients with chronic Keshan disease in Gansu Province.Methods:From March to June 2018, 243 patients with chronic Keshan disease who were treated by self-management treatment outside the hospital in 7 Keshan disease counties in Gansu Province were selected as the survey subjects, and their general demographic and clinical data were collected. At the same time, the survey subjects were given a 6-month self-management treatment, and the self-management scale was used to evaluate the patients' self-management behavior before treatment and 6 months after treatment. The higher the score, the better the patients' self-management behavior. The influencing factors of self-management scale were analyzed by multiple linear regression analysis.Results:Excluding incomplete data, a total of 158 patients with chronic Keshan disease were included in this survey, including 96 males and 62 females. Compared with patients before self-management treatment, the patients' cardiac function classification and work ability improved significantly after 6 months of self-management treatment ( Z = - 4.685, - 5.934, P < 0.05); cardiothoracic ratio decreased (0.61 ± 0.08 vs 0.63 ± 0.09, t = 5.175, P < 0.05); in the cardiac color Doppler ultrasound indexes, left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were increased (0.41 ± 0.11 vs 0.36 ± 0.07, 0.21 ± 0.07 vs 0.18 ± 0.05, t = - 6.504, - 5.391, P < 0.05); the total score of the self-management scale was increased [(53.86 ± 9.29) scores vs (51.46 ± 10.50) scores, t = - 3.696, P < 0.05]. Further analysis of the efficacy of the patients' cardiac function classification showed that the patients' medication compliance was positively correlated with the efficacy of the cardiac function classification ( r = 0.243, P < 0.05). The result of multiple linear regression analysis showed that education level and efficacy of the cardiac function classification were influencing factors of the self-management scale for patients with chronic Keshan disease ( t = 2.466, 2.635, P < 0.05). Conclusion:The self-management treatment for patients with chronic Keshan disease can effectively improve heart function and improve self-management behavior.

Objective:By investigating the cognitive level and influencing factors of heart failure knowledge in patients with chronic Keshan disease, so as to provide reference for improving the cognitive level of disease-related knowledge of the patients with Keshan disease in the targeted manner.Methods:In 2019, 100 patients with chronic Keshan disease were selected in Jingchuan County, Gansu Province. A questionnaire survey was conducted using the heart failure knowledge questionnaire to conduct correlation and regression analysis on the cognitive level of patients' heart failure knowledge and influencing factors.Results:Patients' average score of the heart failure knowledge level was (6.18 ± 2.72) points, and the scoring accuracy rate was 41.20%. The Spearman correlation analysis indicated that heart failure knowledge level was associated with annual family income ( r = 0.363, P < 0.01), the hospitalization times due to heart failure ( r = 0.304, P < 0.01), and the number of family members ( r = 0.240, P < 0.05). The results of multiple linear regression showed that the cognitive level of heart failure knowledge in male patients was higher than that in female patients ( P < 0.05). The more hospitalization times ( P < 0.05) and the more family members ( P < 0.05) were, the higher the cognitive level of heart failure knowledge was. Conclusions:The cognitive level of heart failure knowledge of patients with chronic Keshan disease is generally low. Medical staff should actively make use of the patient's hospitalization time for health education, and pay attention to the health education of family members of the patients.

Objective:To isolate HIV envelope specific monoclonal antibodies (mAbs) from a chronic HIV-1 subtype B′-infected individual with broadly neutralizing activity, and identify their binding activity, epitopes and neutralizing activity.Methods:Single B cell sorting were used to isolate antigen-specific B cells from an HIV-1 infected individual. Variable regions of heavy chain and light chain gene of mAbs were obtained by PCR amplification, and were ligated into the expression vector subsequently. After expression and purification of mAbs, the binding activity and neutralizing ability of mAbs were identified by ELISA and neutralization assay respectively.Results:Seven mAbs were obtained from the HIV-1 infected individual. The heavy chain genes of the mAbs are derived from IGHV1-69*09, IGHV1-08*01, IGHV1-46*01, IGHV4-34*01, IGHV4-61*01 and IGHV3-43*02 germline, SHM rates range from 10.76% to 21.05%. The light chain genes of the mAbs originate from IGKV3-20*01, IGKV1-39*01, IGKV1-NL1*01, IGKV3-15*01 and IGKV1-9*01 germline, SHM rates cover between 6.03% and 18.64%. Gp140 was well recognized by all seven antibodies. 508-B1, 508-C1, 508-C5, 508-D4 and 508-E5 are gp41-directed mAbs. 508-D6 and 508-F1 bind with gp120. 508-D4 and 508-F1 could neutralize pseudoviruses CNE8 and X2278 in Global Panel with IC 50 values of 38.1μg/ml and 41.7 μg/ml, respectively. Conclusions:This study successfully identified seven envelope specific mAbs, among them five mAbs are directed to gp41 region and two mAbs are targeted to gp120. Two mAbs showed limited neutralization spectrum. These mAbs may provide technical reserves for developing HIV-1 diagnostics kit, antibody-related drugs and vaccines.

The study was initiated from a child with diabetes. After we collected his clinical data and traced back his family history of diabetes, a clinical diagnosis of maturity-onset diabetes of the young(MODY)was made. To amplify and sequence the target gene, the genomic DNA was extracted from the anticoagulant blood samples of the patient and his first-degree relatives, revealing a missense mutation(c.779C>T)in exon 4 of hepatocyte nuclear factor-1α in the proband and his father. The above sequencing result confirms the diagnosis of MODY3. During one year follow-up, the proband achieved the strict control of blood glucose with the use of repaglinide and his father got a notable improvement of blood glucose after his drug was shifted to the long-acting sulfonylurea.