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关键词： 牛结节性皮肤病病毒   羊痘病毒   p32基因   原核表达   多克隆抗体  

摘要： 本研究利用原核表达系统对牛结节性皮肤病病毒（LSDV）p32基因N端蛋白进行表达，经免疫特性鉴定后进行多克隆抗体的制备，为研发LSDV血清学检测方法提供基础。根据GenBank中p32基因N端基因序列，构建了p32的体外重组表达质粒，在大肠杆菌中表达并经亲和层析纯化获得重组LSDV-p32N-His蛋白，利用SDS-PAGE对重组蛋白进行鉴定。鉴定后的蛋白一方面以该蛋白作为间接ELISA原料，建立ELISA方法进行特异性和灵敏度测试，进一步对92份田间样品盘进行检测，并与IDvet试剂盒进行符合率比较。另一方面，以该抗原免疫BALB/c小鼠制备多克隆抗体，并分别包被重组抗原和灭活病毒用间接ELISA方法测定多抗效价，利用IFA鉴定多克隆抗体。结果显示，表达的p32N蛋白相对分子质量约为13 kDa，该抗原能够与LSDV和GTPV灭活阳性血清反应，与牛、羊阴性血清和其他血清对照不反应；包被重组抗原用间接ELISA方法对92份田间样品盘进行检测，结果显示该方法与IDvet试剂盒检测的符合率为93.48%；所制备的多抗以重组p32 N蛋白能够检测到1︰5 120 000的血清效价，以灭活病毒为包被抗原检测到血清效价为1︰400；IFA结果显示，该多抗能特异地识别LSDV病毒。本研究结果为深入探索p32蛋白的生物学作用提供了坚实的基础，也为建立LSDV血清学诊断方法、后续筛选单抗及研制亚单位疫苗奠定了基础。

关键词： TNF-Alpha   HERV-K Env   Inflammatory disease   Tissue microarray   Immunofluorescence  

摘要： BackgroundHuman endogenous retroviruses (HERVs) were integrated into the human genome millions of years ago and have since proliferated to comprise about 8% of the human genome. For a long time, HERVs were thought to be remnants of ancient viruses, rendered inactive over the ages. However, recent studies have revealed that HERVs are involved in various diseases, including cancer. Notably, HERVs have been found to play a crucial role in immune responses and inflammatory processes, indicating their significant influence on the regulation of immune-related *** reported in previous reports that HERV-K119 env Knockout (KO) and inflammatory response were associated. In this study, we identified the correlation between inflammatory disease and HERV-K Env and TNF-Alpha protein expression in multiple types of colon disease tissue and lung disease spectrum *** performed Immunofluorescence (IF) using multiple types of colon disease and lung disease spectrum tissue microarray (TMAs) and compared and analyzed the patient clinical data *** a result, we identified that the expression of HERV-K Env and TNF-Alpha proteins in certain colorectal inflammatory diseases and certain lung inflammatory diseases showed specific expression. And through the analysis of the clinical data provided, environmental factors could be *** study demonstrates that the relationship between TNF-Alpha and HERV-K Env expression in inflammation disease and clinical significance of disease tissues.

关键词： Gout   Urate   SNP   IL1RN   Locus  

摘要： ObjectivesGout is caused by the response of the innate immune system to monosodium urate (MSU) crystals. A recent gout GWAS identified a signal of genetic association at a locus encompassing IL1RN-IL1F10. Colocalisation analysis using Genotype Tissue Expression Database (GTEx) eQTL data showed that the signal overlaps with genetic control of IL1RN/IL1F10 gene expression. We assess the functional implications of IL1RN rs9973741, the lead gout-associated *** conducted functional validation of IL1RN rs9973741 in patients with gout and controls. The transcription level of IL1RN/IL1F10 was investigated in unstimulated or MSU-crystal co-stimulated PBMCs. Ex vivo functional assays were performed in PBMCs stimulated with C16 + MSU crystals or LPS for 24 h. Cytokine levels were assessed by *** unstimulated PBMCs, no association of IL1RN rs9973741 (gout risk allele G) to IL1RN expression was observed while IL-1F10 was hindered by low expression at both transcriptional and protein levels. However, G allele carriers showed lower IL1RN expression in PBMCs stimulated with C16/MSU crystal and lower concentrations of circulating IL-1Ra in both controls and gout patients. PBMCs depicted less spontaneous IL-1Ra release in GG homozygous controls and lower IL-1Ra production in response to C16 + MSU crystal costimulation in patients with gout. The G allele was associated with elevated IL-1 beta cytokine production in response to C16 + MSU crystal stimulation in *** gout risk allele G associates with lower circulating IL-1Ra, lower IL-1Ra production in PBMC assays and elevated IL-1 beta production in PBMCs challenged with C16 + MSU crystals but not in unchallenged cells. Our data indicate that the genetic signal that associates with gout at IL1RN-IL1F10 region functions to alter expression of IL-1Ra when stimulated by MSU crystals.

关键词： AKI   IL-6   NAMPT   renal stromal cell   sepsis  

摘要： BACKGROUND AND HYPOTHESIS:Activated macrophages, pivotal for driving the immune response in sepsis, express high levels of CD38. Although the circulating levels of its ligand, CD31, increase in sepsis, the functions of CD38 and its ligation remain elusive. This study aimed to elucidate the impact of CD38 ligation on sepsis using single-cell and single-nucleus RNA sequencing (scRNA-seq and snRNA-seq, respectively) to identify a novel therapeutic target for severe sepsis. METHODS:We performed scRNA-seq analysis of mouse peritoneal immune cells to precisely identify cell types exhibiting increased CD38 expression upon exposure to lipopolysaccharide (LPS). Subsequently, we induced CD38 ligation using a well-established agonistic anti-CD38 antibody in a mouse model of LPS-induced sepsis. We analyzed its pathophysiological effects using kidney snRNA-seq. Finally, we performed histological analysis of septic tissues collected from patients to ensure consistency of our findings between mice and humans. RESULTS:LPS stimulation upregulated CD38 expression in peritoneal macrophages. CD38 ligation significantly exacerbated LPS-induced inflammation in vivo, particularly in the kidneys. Kidney snRNA-seq analysis revealed that CD38 ligation induced interleukin (IL)-6 production in renal stromal cells via nicotinamide phosphoribosyltransferase (NAMPT) signaling originating from CD38-positive macrophages. NAMPT inhibition significantly ameliorated LPS-induced IL-6 production and kidney injury. Histological analysis of human septic tissues demonstrated upregulation of IL6 mRNA and NAMPT in renal stromal cells and CD38-positive macrophages, respectively. CONCLUSION:Our findings elucidate the implications of CD38 ligation in an LPS-induced sepsis model and uncover shared signaling pathways between mice and human sepsis. NAMPT signaling identified in this study may be a novel therapeutic target for mitigating systemic inflammation and kidney injury associated with severe sepsis.

关键词： 肺癌   免疫治疗   肿瘤浸润淋巴细胞   过继性细胞疗法  

摘要： 肺癌是全球癌症相关死亡的主要原因，尽管过去在肺癌治疗方面取得了许多进展，但对于那些不适合靶向治疗及免疫治疗或在一线治疗后出现进展的患者，目前尚缺乏有效的二线治疗方案。基于肿瘤浸润淋巴细胞(tumor-infiltrating lymphocytes,TILs)的过继性细胞疗法(adoptive cell therapy,ACT)，是一种高度个性化的癌症免疫治疗方法，利用患者自身实体瘤微环境的免疫细胞杀死肿瘤细胞。该文综述TILs疗法的基本过程，讨论了当前在肺癌治疗过程中TILs方案的应用以及TILs疗法的局限性和未来发展方向，以期在肺癌治疗过程中获得更广泛的临床应用。

关键词： COVID-19   IL-6   Polymorphism   rs1800795   rs1800796   Severity of disease   Turkish population  

摘要： Coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is exacerbated by cytokine storms, leading to severe inflammation. Interleukin-6 (IL-6) plays a critical role in this process, and variations in its promoter may influence disease severity. This study aims to investigate the relationship between IL6 promoter polymorphisms rs1800795 (G > C) and rs1800796 (G > C) and the severity of COVID-19 in the Turkish population. A total of 332 participants were included: 84 control, 80 with mild COVID-19, and 168 with severe COVID-19. IL6 polymorphisms were genotyped using the restriction fragment length polymorphism (RFLP) method. The genotypes rs1800795 GC (OR = 3.00, 95% CI: 1.669-5.398, p < 0.000), CC (OR = 7.44, 95% CI: 2.899-19.131, p < 0.000), and rs1800796 GC (OR = 2.76, 95% CI: 1.603-4.761, p < 0.000), as well as the alleles rs1800795 C (OR = 3.01, p < 0.000) and rs1800796 C (OR = 1.97, p = 0.002), may be associated with the severity of COVID-19. According to the Jonckheere-Terpstra (J-T) test, the most significant trends that vary linearly with disease severity were observed for D-dimer [J-T = 15.896, Effect size = 0.68 (0.61 to 0.76), p < 0.000] and CRP [J-T = 15.389, Effect size = 0.66 (0.59 to 0.73), p < 0.000]. The distribution of clinical parameters across genotype combinations (rs1800796/rs1800795*) showed that GC/GC* and GC/CC* were linked to a higher risk of severe inflammation, clotting, and organ damage. Additionally, it has been determined that the G-C and C-C haplotypes may be associated with increased severity of COVID-19. The rs1800795 and rs1800796 polymorphisms are linked to COVID-19 severity and could help guide future treatment strategies.

关键词： cisplatin   Roflumilast   Inflammation   apoptosis  

摘要： Purpose: The study aimed to evaluate the effect of roflumilast on modulating TNF-alpha/Caspase mediated cellular signals in cisplatin-induced nephrotoxicity in rats. Methods: The rats (Male Wistar) were divided into five groups: normal control, disease control (cisplatin: 7 mg/kg i.p.), and cisplatin + roflumilast (0.25, 0.5, and 1 mg/kg b.w., p.o.). Cisplatin was administrated to rats on 0 day, and roflumilast treatment was started from the 6th-15th days. Blood and tissue were collected. Tissue was used to measure oxidative stress, such as malondialdehyde, superoxide dismutase, and catalase. Gene expression study involved real-time PCR of key genes linked with inflammation and apoptosis, i.e. Tnf-alpha, Tnfr1, Tnfr2, Fas, Nfkb, Casp3, Casp8, and Nrf2. Findings: Cisplatin showed decreased serum creatinine and urea, high albumin, and total protein. Cisplatin elevated the malondialdehyde and reduced superoxide dismutase and catalase activity. Cisplatin also attributed an overexpression of Tnf-alpha, Tnfr1, Tnfr2, Nfkb, Fas, Casp3, and Casp8, and a decrease in the Nrf2 gene. Roflumilast decreased creatinine and urea and increased albumin and total protein levels. Roflumilast also downregulated the expression of Tnf-alpha, Tnfr1, Tnfr2, Nfkb, Fas, Casp3, and Casp8 and upregulated the Nrf2 gene expression. Conclusion: Roflumilast manifested as a potential reno-protective agent against cisplatin-induced nephrotoxicity.