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关键词： Crohn's disease   Ulcerative colitis   Vitamin D receptor   1,25(OH)(2)D-3   Triglyceride   Colonic mucosal immune  

摘要： The inhibitory effects of the tumor necrosis factor-alpha (TNF-alpha) antibody infliximab (IFX) on colitis are well established. Since IFX dosing is weight-based and associated with various side effects, there is a growing interest in identifying combination therapies that can enhance its efficacy, particularly in overweight inflammatory bowel disease (IBD) patients, to maximize the anti-inflammatory effect while minimizing the required dose. Our research revealed that overweight IBD patients present decreased vitamin D levels in the intestinal epithelium alongside elevated TNF-alpha levels. In mice fed a high-fat diet (HFD) for four weeks, treatment with the vitamin D analog palicalcitol (PAL) reduced lipid synthesis and TNF-alpha production in intestinal epithelial cells (IECs). In a 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced experimental colitis model, PAL treatment mitigated TNF-alpha-induced damage to the intestinal epithelial barrier and reduced the activation of Th1 and Th17 cells in the lamina propria, thereby reducing colitis development in HFD-fed mice. Notably, the combination of IFX and PAL was more effective than IFX alone in treating colitis in these mice. Overall, our findings suggest that vitamin D inhibits TNF-alpha production by reducing lipid synthesis in IECs, thereby enhancing IFX therapy in overweight IBD patients.

关键词： Gastric carcinoma   AFP   HLA-G   PD-L1   HLA- I  

摘要： Alpha-fetoprotein-producing gastric carcinoma (AFPGC) is an aggressive subtype of gastric cancer characterized by a primitive cellular phenotype and poor prognosis. The tumor immunology of AFPGC remains largely unexplored. Given its embryonic-like properties, AFPGC is hypothesized to employ distinct immune evasion strategies, with the oncofetal protein human leukocyte antigen (HLA)-G-a key mediator of maternal-fetal immune tolerance-likely playing a pivotal role. To test this, we assessed the expression of HLA-G, along with other key immune evasion markers, including HLA class I (HLA-I) deficiency and PD-L1 expression, in 39 cases of AFPGC, and compared them with those of 44 Epstein-Barr virus (EBV)-positive, 57 microsatellite instability (MSI), 54 intestinal-type, and 45 diffuse-type gastric carcinomas. HLA-G expression was significantly higher in AFPGCs (71%) than in other subtypes (7-28%;P < 0.001). HLA-I deficiency (>= 1% of tumor cells) was most prevalent in AFPGC (69%), followed by MSI tumors (56%), with lower rates in other subtypes (22-29%). PD-L1 positivity (combined positive score >= 5) was observed in 41% of AFPGCs, lower than in EBV-positive (77%) and MSI tumors (44%), but higher than in intestinal-type (13%) and diffuse-type (9%) carcinomas. Furthermore, CD8-positive T-cell infiltration was found to be lowest in AFPGC compared to the other subtypes. These findings suggest that AFPGC employs multiple immune evasion mechanisms, notably through increased HLA-G expression and HLA-I deficiency, likely linked to its primitive cellular phenotype and reactivation of immunogenic oncofetal antigens. Such immune evasion features may underlie the aggressiveness of AFPGC and present promising targets for immunotherapeutic interventions.

关键词： 复发性流产   白细胞介素-6   干扰素-γ   妊娠结局   相关性  

摘要： 目的:探讨血清白细胞介素-6(IL-6)、干扰素-γ(IFN-γ)水平与复发性流产(RSA)患者妊娠结局的相关性分析.方法:选取2023 年 11 月至 2024 年 10 月本院收治的 122 例RSA患者作为观察组,纳入同期 100 例无流产史女性作为对照组,均为孕早期.对比两组的血清IL-6、IFN-γ表达水平,对观察组患者跟踪随访,根据妊娠结局将观察组分为妊娠结局良好组(n=75)和不良组(n=47),对比两组的一般资料及IL-6、IFN-γ表达水平,并采用Logistic回归分析复发性流产患者妊娠结局的影响因素,并采用Spearman分析IL-6、IFN-γ与RSA患者妊娠结局的关系.结果:观察组IL-6 低于对照组,IFN-γ高于对照组(P<0.05);妊娠不良组的β-HCG、孕酮、IL-6 低于良好组,IFN-γ高于良好组(P<0.05);Logistic回归分析显示,β-HCG(OR=-0.687,95％CI:0.387～0.619)、孕酮(OR=-0.577,95％CI:0.417～0.707)、IL-6(OR=-0.614,95％CI:0.418～0.664)升高是RSA患者妊娠结局不良的保护因素,IFN-γ(OR=1.131,95％CI:1.274～4.924)升高是危险因素(P<0.05).Spearman分析显示,血清IL-6 与RSA患者妊娠结局良好呈正相关(r=0.618,P<0.001),血清IFN-γ与RSA患者妊娠结局良好呈负相关(r=-0.634,P<0.001).结论:血清IL-6 在RSA患者妊娠结局不良中呈低表达,IFN-γ呈高表达;IL-6 表达水平升高是RSA患者出现不良妊娠结局的保护因素,IFN-γ表达水平升高是危险因素,且血清IL-6,IFN-γ水平与复发性流产患者妊娠结局存在明显相关性.

摘要： Familial adenomatous polyposis (FAP) is an inherited gastrointestinal syndrome associated with duodenal adenoma formation. Even among carriers of the same genetic variant, duodenal phenotypes vary, indicating that additional factors, such as the local immune system, play a role. We observe an increase in duodenal IL-17A(+)NKp44(-) innate lymphoid type 3 cell (ILC3) in FAP, localized near the epithelium and enriched in adenomas and carcinomas. Elevated IL1B, IL23A, and DLL4 transcript levels correlate with IL-17A(+)NKp44(-)ILC3 accumulation, and in vitro studies with duodenal organoids confirmed this relationship. Bulk RNA sequencing reveals upregulated Reactive oxygen species (ROS)-inducing enzymes DUOX2 and DUOXA2 in FAP adenomas. IL-17A-stimulated FAP organoids show increased DUOX2/DUOXA2 expression, Duox2 protein, and ROS production, leading to DNA damage, suggesting a mechanism by which these immune cells promote tumorigenesis. These findings suggest IL-17A(+)NKp44(-)ILC3s may contribute to a local environment that makes the epithelium more submissive for oncogenic transformation in FAP.

关键词： Bioanalysis   EliSpot   FluoroSpot   GLP   Validation  

摘要： The enzyme-linked immunosorbent spot (EliSpot) assay and its fluorescence-based version, FluoroSpot, are sensitive immunoassays commonly used to quantify antigen-specific T and B lymphocytes and other immune cells in peripheral blood or homogenized tissues. Due to their high sensitivity, these assays are popular in clinical trials to evaluate the efficacy of immunotherapy and vaccines, which involve a high level of scrutiny to ensure valid study results. Besides industry consensus white papers and other research publications, there is no formal guidance for the industry on how to validate EliSpot and FluoroSpot assays to ensure their accurate performance for immune monitoring in clinical trials. Herein, we describe a comprehensive in vitro study using healthy human donor peripheral blood mononuclear cells (PBMCs) and model antigens to validate a double-color FluoroSpot assay for monitoring antigen-specific lymphocytes by detecting and quantifying IFN-gamma and IL-2-producing lymphocytes. Validation parameters, acceptance criteria set-up, and assay limits-limit of detection (LOD), minimum positive control response, lower and upper limits of quantification (LLOQ and ULOQ)-were determined, and assay performance was demonstrated by assessing precision, specificity, linearity, and robustness. In addition, an inter-laboratory comparison demonstrated concordance between assay results from two laboratories. In summary, this study outlines a robust approach to EliSpot and FluoroSpot validation and demonstrates that the IFN-gamma/IL-2 FluoroSpot assay is suitable for the reliable detection of antigen-specific immune responses from PBMC samples across laboratories and meets the current regulatory requirements for bioanalytical method validation.

关键词： children   severe community-acquired pneumonia   IL-1 family   pleural effusion   Haemophilus influenzae  

摘要： Background: To investigate the evaluative role of interleukin (IL)-1 family cytokines in bronchoalveolar lavage fluid (BALF) among children with severe community-acquired pneumonia (SCAP) and identify cytokines with clinical relevance for pediatric SCAP. Methods: Children with SCAP hospitalized at Shenzhen Children's Hospital (2019-2020) were studied. IL-1 family cytokines in the BALF were measured via CBA or ELISA. These cytokines included nine IL-1 family members (IL-1 alpha, IL-1 beta, IL-1Ra, IL-33, IL-18, IL-37, IL-36 alpha, IL-36Ra, and IL-38) and two receptors (sST2 and IL-18BP). The ratio of proinflammatory cytokines to anti-inflammatory cytokines was analyzed. Results: In the BALF of children with SCAP complicated with pleural effusion (PE), the levels of IL-18, the IL-18/IL-38 ratio, and the IL-33 level were significantly elevated (P < 0.05). Furthermore, the receiver operating characteristic (ROC) curve indicated that these three markers have strong predictive efficacy for diagnosing SCAP complicated with PE. The levels of members of the IL-1 family, including IL-1 alpha, IL-1 beta, IL-1Ra, IL-18, and IL-33, and their associated ratios significantly differed across different pathogen groups (P < 0.05). IL-36 alpha and the IL-36 alpha/IL-38 ratio differed significantly between the Haemophilus influenzae (Hi)-positive and -negative groups (P < 0.0001 and 0.0048), with lower levels in the Hi-positive group. Conclusion: IL-18, IL-33, and IL-38 in BALF may serve as effective markers for predicting the development of PE in pediatric SCAP patients. Additionally, respiratory tract colonization by Hi may diminish the production of specific proinflammatory cytokines, including IL-18, IL-33, and IL-36 alpha, during SCAP.

关键词： 绵羊   卵母细胞   体外成熟   体外胚胎生产  

摘要： 该研究在绵羊卵母细胞体外成熟液中添加40 ng/mL成纤维生长因子2 (FGF2)、20 ng/mL白血病抑制因子(LIF)和20 ng/mL类胰岛素生长因子1 (IGF-1)细胞因子组合(FLI),设置对照组(常规成熟液)与试验组(FLI添加组),体外成熟后进行统一体外受精和胚胎培养,统计胚胎卵裂率和囊胚率,探究体外成熟液添加FLI对绵羊体外胚胎发育效率的作用。使用免疫荧光染色检测囊胚的H3K4me3水平,使用TUNEL染色检测囊胚细胞凋亡水平,使用荧光定量PCR检测多能性基因和凋亡基因表达水平,探究体外成熟液添加FLI对绵羊体外胚胎质量的影响。结果表明:FLI组卵裂率和囊胚率均高于对照组,但均无显著差异(P>0.05)。不同处理对囊胚的总细胞数、H3K4me3水平、凋亡细胞比率无显著影响(P>0.05)。然而,与对照组相比,FLI组囊胚中凋亡相关基因(P53、CASPASE3和BAX)表达水平显著降低(P<0.05),多能性基因OCT4表达显著升高(P<0.05)。可见,绵羊卵母细胞体外成熟液添加FLI虽不影响胚胎发育率,但可能对胚胎质量有提高作用。

关键词： 坤宁颗粒   异常子宫出血   子宫动脉血流   PBAC评分   子宫内膜厚度  

摘要： 目的:探讨坤宁颗粒联合口服避孕药对异常子宫出血患者子宫动脉血流及细胞因子水平的影响.方法:选择 2021 年 3月至 2024 年 3 月收治的 80 例功能失调性子宫出血患者为研究对象并行前瞻性研究,受试者根据随机数字表法予以分组,各 40 例,对照组(口服避孕药)与研究组(坤宁颗粒+口服避孕药),对比两组患者治疗效果与各细胞因子水平变化、子宫动脉血流、子宫出血与月经失血图(PBAC)评分及子宫内膜厚度变化以及用药安全性.结果:研究组总疗效比对照组更高(P<0.05);研究组相关治疗时间指标均较对照组短(P<0.05);组间各血清细胞因子水平相比,治疗后研究组PGE2 与Ang-Ⅱ水平较对照组低,β-EP水平较对照组高(P<0.05);组间PI、RI及S/D相比,治疗后研究组较对照组低(P<0.05);组间子宫出血与PBAC评分及子宫内膜厚度相比,治疗后研究组子宫出血评分与子宫内膜厚度较对照组低,PBAC评分较对照组高(P<0.05);组间用药安全性相比,数据评比结果显示无差异性(P>0.05).结论:在异常子宫出血治疗中联合应用坤宁颗粒与口服避孕药可有效调节各细胞因子水平,改善子宫动脉血流及子宫内膜厚度,疗效显著且安全,对促进患者病情康复具有积极意义.

关键词： genetics   population diversity   precision medicine   type 1 diabetes   type 2 diabetes  

摘要： Aims: We have reported that a 9SNPs type 1 diabetes (T1D) Genetic Risk Score (GRS) developed from European data had a lower power in Indians to distinguish T1D from type 2 diabetes (T2D). We explore the performance of an improved (67SNPs) T1DGRS and also the potential reasons for lower discriminative ability to classify types of diabetes in Indians. Methods: We studied the discriminative ability of a 67SNPs European T1DGRS in 611 clinically diagnosed T1D and 1153 T2D patients, and 321 non-diabetic controls, using receiver operating characteristic (ROC) area under the curve (AUC). We also compared the frequency and effect sizes of HLA risk haplotypes between Indians and Europeans. Results: The T1DGRS was discriminative of T1D from T2D and controls. However, the ability is lower in Indians than Europeans (AUC, Europeans 0.92, Indians all T1D 0.83, AA-positive 0.86). The T1DGRS was higher in AA-positive than in AA-negative persons [13.01 (12.79-13.23) vs. 12.09 (11.64-12.56)], p < 0.0001. The association of common HLA-DQA1 similar to HLA-DQB1 haplotypes was broadly similar;however, important differences were noted in the frequency, direction and magnitude of effect for some haplotypes between Indians and Europeans. Conclusions: We confirm broad applicability of European 67SNPs T1DGRS to Indian T1D persons. However, differences in HLA allele frequencies, magnitude and directional differences reduced the predictive value. Our results stress the need to generate ancestry-specific GRS, which we plan to do in the near future.

关键词： peginterferon alfa-2b   chronic hepatitis B virus infection   liver function   immune factors   prognosis  

摘要： Aims/Background Hepatitis B virus (HBV) infection poses a challenge to global healthcare. Peginterferon alfa-2b (PEG-IFNa-2b) is an effective treatment for HBV infection. This study aimed to explore the efficacy of PEG-IFNa-2b combined with entecavir in the treatment of HBV infection, its effect on liver function and immune factors, and the risk factors affecting the prognosis of patients with HBV infection. Methods The clinical data of 184 patients with HBV infection who were treated at Jinhua Central Hospital from January 2021 to January 2024 were collected for retrospective analysis. Patients were divided into a control group (not receiving antiviral treatment, n = 34), a standard treatment group (receiving entecavir, n = 85), and a combination treatment group (PEG-IFNa-2b and entecavir, n = 65) according to the treatment approach. Treatment efficacy, liver function indicators (albumin [ALB], alanine aminotransferase [ALT], and aspartate aminotransferase [AST]), immune factor indexes (tumour necrosis factor alpha [TNF-a] and interferon gamma [IFN-'y]), hepatitis B surface antigen [HBsAg] and HBV DNA levels were compared among the three groups. All patients were followed up after treatment. According to their prognosis, the patients were divided into good prognosis group (n = 118) and poor prognosis group (n = 66). Logistic regression analysis was performed to explore the risk factors affecting the prognosis of HBV patients. Results The efficacy in the combination treatment group was higher (92.31%) than that in the control group (8.82%) and the standard treatment group (78.82%) (p G 0.05). After treatment, the HBsAg and HBV DNA levels were decreased in the standard treatment and combination treatment groups (p G 0.05). Compared with the control and standard treatment groups, the combination treatment group exhibited significantly lower HBsAg and HBV DNA levels after treatment (p G 0.05). Besides, the combination treatment group had lower ALT and AST levels (p G 0