课程文献中心 更多>>

关键词： Multiple sclerosis   Cognitive disability   IL-6 rs1800795 G > C   EBV  

摘要： Multiple Sclerosis (MS) is a chronic immune-mediated neurological disorder frequently accompanied by cognitive impairment, which affects up to 60% of patients and is associated with faster disease progression and greater disability. Interleukin-6 (IL-6), a key proinflammatory cytokine involved in neuroinflammation, has been implicated in MS pathogenesis, and the rs1800795 (-174 G>C) single nucleotide polymorphism (SNP) in the IL6 gene may influence disease susceptibility and clinical severity. This study investigated the association between the IL6 rs1800795 polymorphism and clinical outcomes in Epstein-Barr virus (EBV)-positive MS patients, with a particular focus on cognitive dysfunction. A case-control design was employed, including 300 participants: 150 EBV-positive MS patients and 150 matched healthy controls. Genotyping was performed using TaqMan-based PCR, and clinical data such as disability status, disease progression, and cognitive performance were analyzed. The CC genotype was significantly more frequent in MS patients and was associated with a higher risk of severe disability (OR = 6.11, p = 0.0004), faster disease progression, and increased likelihood of cognitive impairment. These findings suggest that the IL6 rs1800795 polymorphism, particularly the CC genotype, contributes to MS susceptibility and adverse clinical outcomes. IL6 genotyping may hold promise as a predictive tool for disease progression and cognitive decline in EBV-associated MS, offering insights for more personalized therapeutic strategies.

关键词： interleukin 6   endothelial cell dysfunction   severe pneumonia   inflammation   allo-HSCT   acute lung injury  

摘要： Background Severe pneumonia after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is associated with high mortality. Given that cytokines, including interleukin 6 (IL-6), play a critical role in immune-mediated organ injury in patients with severe coronavirus disease 2019 (COVID-19), we hypothesized that cytokines may also contribute to the pathogenesis of severe pneumonia after allo-HSCT. This study aimed to investigate the role of IL-6 in severe pneumonia after allo-HSCT and explore its underlying *** Serum cytokine levels were prospectively measured in patients with severe and nonsevere pneumonia following allo-HSCT. A mouse model of acute lung injury (ALI) and in vitro experiments using primary murine pulmonary microvascular endothelial cells (PMVECs) were conducted to assess the effects of IL-6 blockade, the mechanism of IL-6 in ALI, and immune-induced *** Serum IL-6 and soluble IL-6 receptor (sIL-6R) levels were higher in the severe pneumonia group than in the nonsevere group and were associated with disease progression. In a mouse model, preventive IL-6 blockade reduced ALI and improved survival. In vitro, the IL-6 trans-signaling complex caused more severe damage to mouse PMVECs than the classical signaling pathway. Soluble glycoprotein 130 and ruxolitinib effectively blocked the JAK1/STAT3 pathway activated by IL-6 trans-signaling in mouse PMVECs and reduced downstream inflammatory *** IL-6 levels were elevated in patients with severe pneumonia after allo-HSCT and were linked to disease progression. This injury may be driven by the IL-6/sIL-6R/JAK1/STAT3 pathway. This preliminary study suggests that targeting the IL-6 trans-signaling pathway may be a promising therapeutic approach for severe pneumonia/ALI following allo-HSCT. Interleukin 6 was elevated in severe pneumonia after allo-HSCT and plays a role in lung injury via the IL-6/sIL-6R/JAK1/STAT3 pathway.

关键词： Carbonic anhydrase III (CAIII)   Tumor necrosis factor (TNF-alpha)   Colon carcinoma   Osteosarcoma   Gene Regulation  

摘要： The appropriate acid-base balance in organisms is maintained by Carbonic Anhydrase proteins (CAs), which have hydratase activity and regulate intracellular pH. CAs are required for both physiological and pathophysiological processes such as cancer development, and there are differences in their expression profiles in different cancer types. Some members of the CA family like CAIX and CAXII have been suggested as potential cancer biomarkers in various studies. CAIX has been proposed as a possible biomarker for hypoxic colorectal carcinoma. Expression of CAIII, another member of the CA family, was also found to be reduced by TGF-beta via the MAPK and PI3K signaling pathways in human colon cancer cells. Additionally, not much is known regarding the interaction between TNF-alpha, inflammation-related cytokine, and CAIII in colorectal carcinoma (CRC). This study investigates the effect of TNF-alpha on CAIII expression in different cancer cells, namely HT-29 and Saos-2. CAIII mRNA expression was analyzed by qRT-PCR at both late (24, 48, and 72 h) and early time points (1, 3, and 6 h). Western blot analysis was also used to confirm the reducing effect of TNF-alpha at the CAIII protein level. To analyze the transcriptional regulation of CAIII by TNF-alpha, CAIII promoter constructs were transiently transfected to HT-29 and Saos-2 cells, and transcriptional activities of truncated promoter constructs were analyzed with luciferase/SEAP activities. 500U/mL TNF-alpha led to a drastic decrease at 24 and 48 h time points at CAIII mRNA level. Western blot analysis showed that this decreasing effect of 500 U/mL TNF-alpha at 24 h, 48 h, and 72 h resulted in a reduction of the CAIII protein level by 0.5 times. P1 (- 939/+ 86), P2 (- 699/+ 86), P3 (- 236/+ 86), and P4 (- 108/+ 86) CAIII promoter constructs were transiently transfected to HT-29 cells, P4 (- 108/+ 86) promoter basal activity is greater than the other promoter constructs. Transcriptional activity of all CAIII promoter co

摘要： The infrapatellar fat pad (IPFP) acts as a bioactive reservoir, secreting proinflammatory cytokines that orchestrate both local and systemic inflammatory cascades. Despite its emerging role in knee osteoarthritis (OA) pathophysiology, the molecular and cellular mechanisms driving IPFP-mediated disease progression remain a critical gap in mechanistic understanding. 12-week-old male C57BL/6 mice underwent either destabilization of the medial meniscus (DMM) surgery or Sham surgery. Here, we find that the extreme sensitivity of IPFP makes it prone to act as a reservoir of inflammatory factors, which may indiscriminately disrupt the stability of its surrounding tissues. We further ascertain the role of IL-6 in initializing fibrosis in IPFP at early stage of OA and modulating osteopontin (OPN) secretion that cascades cartilage deterioration. Notably, removal of the IPFP in DMM mice reverses the abnormal functions of the knee joint. Compromising the progress of fibrosis by intra-IPFP injection of siRNA Cd61 or inhibition of OPN expression can drastically ameliorate cartilage deterioration. Our findings elucidate a pivotal role for IL-6 in instigating fibrotic remodeling within the IPFP during early-stage OA, concurrently regulating OPN secretion to propagate cartilage matrix degradation. This study thus establishes a conceptual framework for therapeutic intervention by targeting the IL-6/OPN signaling axis in the IPFP during OA initiation, offering a promising strategy to disrupt disease progression.

关键词： Interleukin-25   Induced sputum   Childhood   Asthma   Clinical features   Airway inflammation   Anti-asthma treatment  

摘要： Background: Asthma is a chronic respiratory disease characterized by reversible airway obstruction and persistent airway inflammation, presenting as a highly heterogeneous disorder in children. Further understanding of its complexity is essential to identify applicable biomarkers and targeted therapies. Interleukin-25 (IL-25) has been shown to play a critical role in the pathogenesis of asthma. Methods: To investigate the association between IL and 25 expression and clinical characteristics, we enrolled 46 children with asthma (age 6-17 years) and 15 age-matched healthy controls. Asthma patients were stratified into Group A (untreated, n = 24) and Group B (treatment-controlled, n = 22). IL-25 protein levels in serum and IL-25 mRNA in induced sputum were quantified using ELISA and PCR, respectively. Results: No significant intergroup differences existed in age (P = 0.32), sex (P = 0.67), or BMI (P = 0.144). IL-25 mRNA in sputum was significantly elevated in both groups versus controls (P <0.001 in Group A and P < 0.05 in Group B). Sputum IL-25 protein levels were higher in Group A versus controls (P < 0.001) and Group B (P < 0.05). IL-25 mRNA expression in sputum was significantly higher in Group A (without anti-asthma drugs) compared to Group B (with controlled asthma treated with anti-asthma drugs) (P < 0.05 in both induced sputum and blood). Furthermore, IL-25 mRNA expression correlated with CRP (P = 0.007), FeNO (P = 0.04), FEV1/FVC (%) (P = 0.01), induced sputum eosinophil count (%) (P = 0.03), disease severity (P = 0.042), and anti-asthma treatment (P < 0.05). Notably, IL-25 levels in induced sputum decreased significantly at both molecular and gene levels following anti-asthma treatment, suggesting its potential as a biomarker for evaluating treatment efficacy and asthma control. Conclusion: IL-25 expression in induced sputum may serve as a reliable biomarker in children with bronchial asthma, though further large-scale studies are needed to confirm these find

关键词： Fourier transform infrared spectroscopy  

摘要： The present work reports a green approach for the development of a new electrochemical sensor based on a pectin and ionic liquid composite (PC/IL) for sensitive detection of triclopyr (TCP), which is a useful pesticide. The fabricated sensor was characterized using microscopic (Scanning Electron Microscopy (SEM)), spectroscopic (Energy Dispersive X-ray (EDX) spectroscopy, ultraviolet-visible (UV-vis) spectroscopy, X-ray diffraction (XRD), Fourier Transform Infrared (FTIR) spectroscopy, Electrochemical Impedance Spectroscopy (EIS)), and cyclic voltammetric methods. The sensor displayed a large surface area, high conductivity, and rich porosity, which contributed to achieving high electrochemical performances toward sensing applications. Experimental parameters, including pH, solvent, concentration, frequency, and amplitude, were optimized for the detection of TCP at PC/IL@GCE using voltammetric techniques. The developed sensor exhibited high sensitivity, selectivity and reproducibility for TCP detection with the detection limit (LOD) and quantification limit (LOQ) as low as 223 pg mL-1 and 745 pg mL-1, respectively. The electrode also exhibited satisfactory recovery in the range of 99.64-101.92%, 99.44-100.87%, and 99.02-101.40% for the detection of TCP in real samples of human serum, tomato, and apple, respectively. Additionally, the environmental sustainability of the developed sensor was assessed, demonstrating a 92% greener aspect.

关键词： Human induced pluripotent stem cell   Gene engineering   Natural killer cell   Solid tumor   Immunotherapy   Off-the-shelf product  

摘要： BackgroundStudies of chimeric antigen receptor (CAR)-T and -Natural killer (NK) cells have shown promising results in treating hematological malignancies. However, there are still obstacles to effectively treating solid tumors. These include the challenges of CAR-T cell homing and infiltration, the presence of immunosuppressive microenvironments, and the potential for antigen escape in solid tumors. To overcome the known limitations of immune cell therapy, we engineered human induced pluripotent stem cell (hiPSC)-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D-DAP10 *** introduced the six genes, CCL19, CCR2B, FCGR3A (CD16), IL-15, KLRK1 (NKG2D), and HCST (DAP10), which were controlled under human EF1a promoter, into hiPSCs using the piggyBac system and differentiated them into NK cells. We evaluate the antitumor function, including killing activity, antibody-dependent cytotoxicity, migration ability, and recruitment of dendritic cells. In addition, in vivo antitumor activity was determined by using an orthotopic lung cancer mouse *** gene-engineered hiPSCs expressed all six transgenes, showed normal karyotypes, and were able to differentiate into CD56+ NK cells. The gene-engineered hiPSC-derived NK (eNK) cells showed improvement in viability without additional cytokine supplement in vitro and in vivo. Overexpression of NKG2D complex and high-affinity CD16 enhanced the antitumor function of the eNK cells. Forced expression of CCR2B enhanced eNK cell tumor infiltration. Forced expression of CCL19 endowed the eNK cells with the ability to recruit dendritic cells. We found that the eNK cells were able to lyse HLA-E-expressing tumor cells, but not normal human cells. Moreover, eNK cells demonstrated superior anti-tumor activity in an orthotropic lung cancer mouse *** proof-of-concept studies demonstrate the promise of our eNK cells as a novel adoptive cell therapy product for the treatment of sol

关键词： CD318   单克隆抗体   制备   鉴定  

摘要： 目的 制备CD318特异性单克隆抗体并对其特异性、亲和力及在免疫学检测中的应用进行鉴定，为 CD318的抗体药物研发奠定基础。方法 表达并纯化CD318蛋白，作为抗原免疫小鼠，筛选抗血清效价较高的小鼠，经过细胞融合和单克隆筛选，制备CD318的特异性单克隆抗体。对获得的单克隆抗体的特异性、亲和力及其在免疫学实验中的应用进行检测。构建靶向CD318/CD3双特异性抗体，检测其对T淋巴细胞杀伤功能影响。结果 获得13株单克隆抗体，其中效价最好、特异性最强和用途最广泛的杂交瘤细胞株的克隆号为13-8-G2，抗体为IgG1，轻链为kappa型；轻链可变区大小为318 bp，重链可变区大小为357 bp，亲和常数约为7.68×109，可分别在流式细胞术和免疫荧光中用于对CD318的特异检测，用获得的CD318单克隆抗体可变区构建的靶向CD318/CD3双特异性抗体，可增强T淋巴细胞杀伤功能。结论 成功制备并获得了亲和力高、特异性好的CD318单克隆抗体，为CD318靶点的成药性抗体开发奠定了基础。

关键词： Human rhinovirus   Interleukin-23   T helper 17  

摘要： Background The inflammatory response is critical in Human Rhinovirus (HRV) infection-related acute asthma attacks, but its biological mechanisms remain unclear. The interleukin-23 (IL-23)/T helper 17 (Th17) pathway is a key inflammatory pathway that regulates abnormal inflammatory responses. Therefore, this study aims to investigate the role of the IL-23/Th17 pathway in the pathogenesis of wheezing in children with HRV infection by examining the expression of signaling molecules within this pathway. Method Seventy-five pediatric patients hospitalized with HRV infection and 21 healthy children (as a control group) were randomly enrolled from Hangzhou Children's Hospital between July 2022 and November 2023. HRV-infected patients were stratified into wheezing (n = 42) and non-wheezing (n = 33) groups based on wheezing symptoms. Nasopharyngeal swabs and peripheral venous blood samples were collected on admission. An additional set of peripheral venous blood samples was collected from 22 children in the wheezing group during their recovery phase (day 7 after symptom onset). Th17 cell percentages in peripheral blood lymphocytes were determined using flow cytometry. ELISA was utilized to measure the serum levels of IL-23 and IL-17 and the expression of IL-23R, JAK2, p-JAK2, STAT3, and p-STAT3 in peripheral blood mononuclear cells. Comparisons of expression levels were made among the wheezing, non-wheezing, and control groups, as well as between the acute and recovery phases in the wheezing group. Results Children with wheezing due to HRV infection exhibited significantly higher levels of IL-23 and IL-17 compared to the non-wheezing and control groups (p < 0.05). No significant differences were observed in the expression of other molecules in the IL-23/Th17 pathway across the groups (p > 0.05). During the recovery phase, the levels of IL-23 and IL-17 significantly decreased compared to the acute phase (p < 0.05). Conclusion IL-23 and IL-17 significantly contribute to the de

关键词： 猫星状病毒   衣壳蛋白   原核表达   多克隆抗体   间接酶联免疫吸附试验  

摘要： 为了建立猫星状病毒（FAstV）的血清抗体检测方法，本研究从猫粪便拭子中筛查并克隆FAstV ORF2基因。通过原核表达技术成功表达了衣壳蛋白截短体CapA的重组蛋白，以纯化后的CapA重组蛋白作为包被抗原，建立了FAstV血清抗体间接ELISA检测方法，并对临床血清样本进行检测。结果显示：CapA蛋白在*** BL21（DE3）中以可溶性蛋白形式表达其大小约65 kDa；免疫新西兰大白兔制备的多克隆抗体经间接免疫荧光试验验证，可特异性识别FAstV。建立的间接ELISA方法经系统优化，确定最佳条件为：10 μg/mL抗原包被（37℃孵育2 h），1﹕100血清稀释（37℃ 1 h），1﹕20 000 HRP-二抗（37℃ 45 min）。该方法具有高灵敏度（检测限1﹕3 200）和特异性（与猫杯状病毒、猫冠状病毒等无交叉反应），重复性良好（批内批间变异系数均小于10%）。用该方法对广西217份猫血清样本进行检测，结果显示，FAstV抗体的阳性率为11.06%，证实了该检测体系的实用价值。本研究首次报道的FAstV CapA蛋白多抗及ELISA检测方法为FAstV的分子机制研究和流行病学监测提供了重要技术支撑。