A DNA fragment encoding human interleukin 4 was obtained by PCR from pORF-hIL4 plasmid. The amplified fragment was inserted into prokaryotic expression vector PQE60 and recombinant protein was expressed in E. Coli M15 by adding isopropyl-beta-D-thiogalactoside (IPTG). The hIL-4 protein was present as insoluble inclusion bodies in the bacterial extract. After denaturation of inclusion bodies with 5 mol/L guanidine hydrochloride, the supernate was diluted to get renaturized. Then dialysis and Ni chelating chromatography were used for purification. TF-1 proliferation assay of recombinant human interleukin 4 was performed, and then rhIL-4 was fit to be used for proliferation of human dendritic cells from monocyte in vitro.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Inclusion Bodies ; metabolism ; Interleukin-4 ; biosynthesis ; genetics ; Protein Folding ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

[摘 要] 近年来，随着对肿瘤抗原和抗肿瘤免疫反应机制的深入认识，肿瘤治疗性疫苗发展迅速，有望成为临床肿瘤治疗的重要手段。同时，随着新抗原的筛选、疫苗设计、疫苗递送系统、佐剂等关键技术的突破，进一步加速了这一领域的发展。在产品研发方面，众多国际大型的医药企业和新兴的生物科技公司正在布局不同的肿瘤治疗性疫苗项目，多个肿瘤治疗性疫苗获批上市，但临床效果欠佳。尽管目前肿瘤治疗性疫苗大多处于临床前和临床试验阶段，但展现出了良好的临床应用前景和市场价值。本文论述了国内外肿瘤治疗性疫苗的研发现状（主要聚焦当前发展迅速的个性化新抗原疫苗、DC疫苗和mRNA疫苗），总结了目前所面临的挑战并展望了其发展前景，为未来肿瘤治疗性疫苗的研究和产品研发提供了新思路。

OBJECTIVETo observe the difference in the clinical therapeutic effects on bronchial asthma in children of different body constitutions treated withplaster andplaster.

METHODSOne hundred and twenty-two children of bronchial asthma were divided into three groups according to TCM body constitutions, 42 cases in thedeficiency constitution group, 40 cases in thedeficiency constitution group and 40 cases in the phlegm damp constitution group. From 2011 to 2013, the acupoint plaster was applied to all of the children in the three groups during the dog days and the third nine-day period after the winter solstice each year. The average attack frequency and onset days of bronchial asthma and relevant immune function indicators were observed during treatment and 1 year after treatment in the children and the therapeutic effects were evaluated.

RESULTS①In 2014, the acute attacks of bronchial asthma were (1.2±0.9) times and (1.4±0.4) times in thedeficiency constitution group and thedeficiency constitution group, all lower than (3.0±0.5) times in the phlegm damp constitution group (both<0.05) separately. ②After treatment, in thedeficiency constitution group anddeficiency constitution group, the values of IgG, IgA and IgM were all increased as compared with those before treatment (all<0.05). ③The total effective rate was over 95% in the children of the three groups. The clinical control rates in thedeficiency constitution group and thedeficiency constitution group were higher apparently than that in the phlegm damp constitution group, indicating the significant difference statistically (both<0.05).

CONCLUSIONSThe combined treatment ofplaster andplaster are effective on bronchial asthma in the children of different body constitutions. The therapeutic effects for thedeficiency constitution and thedeficiency constitution are more apparent than that for the phlegm damp constitution.

BACKGROUND: Women comprise more than half of people living with human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) worldwide and incomplete immune recovery and metabolic abnormalities affect them deeply. Studies of HIV antiretroviral therapy (ART) have a low female representation in China. We aimed to investigate immune reconstitution and metabolic changes of female HIV-positive cohort in China longitudinally. METHODS: HIV-positive women who initiated ART from January 2005 to June 2021 and were followed up regularly at least once a year were included in this study. Immunological indicators (cluster of differentiation 4 [CD4] counts and CD8 counts), viral load (VL), and metabolic indicators were collected at follow-up. All data were collected from the China Disease Prevention and Control Information System (CDPCIS). VL was tested half a year, 1 year after receiving ART, and every other year subsequently according to local policy. CD4/CD8 ratio normalization was considered as the primary outcome and defined as a value ≥1. Incidence rate and probability of CD4/CD8 ratio normalization were estimated through per 100 person-years follow-up (PYFU) and Kaplan-Meier curve, respectively. Multivariate Cox regression was used to identify independent risk factors associated with CD4/CD8 ratio normalization. We further studied the rate of dyslipidemia, hyperuricemia, diabetes, liver injury, and renal injury after ART initiation with the chi-squared tests or Fisher's exact probability tests, and a generalized estimating equation model was used to analyze factors of dyslipidemia and hyperuricemia. RESULTS: A total of 494 female patients with HIV/AIDS started ART within 16 years from January 2005 to June 2021, out of which 301 women were enrolled with a median duration of ART for 4.1 years (interquartile range, 2.3-7.0 years). The overall incidence rate of CD4/CD8 ratio normalization was 8.9 (95% confidence interval [CI], 7.4-10.6) per 100 PYFU, and probabilities of CD4/CD8 normalization after initiating ART at 1 year, 2 years, 5 years, and 10 years follow-up were 11.7%, 23.2%, 44.0%, and 59.0%, respectively. Independent risk factors associated with CD4/CD8 normalization were baseline CD4 cell counts <200 cells/μL, CD8 counts >1000 cells/μL, and more than 6 months from the start of combined ART (cART) to first virological suppression. Longitudinally, the rate of hypercholesterolemia (total cholesterol [TC]) and high triglyceride (TG) showed an increasing trend, while the rate of low high-density lipoprotein cholesterol (HDL) showed a decreasing trend. The rate of hyperuricemia presented a downtrend at follow-up. Although liver and renal injury and diabetes persisted during ART, the rate was not statistically significant. Older age and protease inhibitors were independent risk factors for increase of TC and TG, and ART duration was an independent factor for elevation of TC and recovery of HDL-C. CONCLUSIONS This study showed that women were more likely to normalize CD4/CD8 ratio in comparison with findings reported in the literature even though immune reconstruction was incomplete.
Humans ; Female ; CD4-CD8 Ratio ; HIV ; Immune Reconstitution ; Hyperuricemia/drug therapy* ; HIV Infections/drug therapy* ; Acquired Immunodeficiency Syndrome/drug therapy* ; Anti-Retroviral Agents/therapeutic use* ; Cholesterol ; Viral Load ; CD4 Lymphocyte Count ; Anti-HIV Agents/therapeutic use*