七氟烷吸入麻醉对单肺通气患者血清IL-6、IL-10、MIP-2、SP-D及HMGB1水平的影响

目的:探讨七氟烷吸入麻醉对单肺通气患者血白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、巨噬细胞炎性蛋白-2(MIP-2)、人肺表面活性特异蛋白(SP-D)及高迁移率族蛋白1(HMGB1)水平的影响。方法:选择2014年9月~2016年9月于我院行单肺通气的患者108例,参照抽签法分为两组,每组各54例。对照组行常规麻醉,实验组采用七氟烷麻醉,比较两组通气前后血清IL-6、IL-10、MIP-2、SP-D、HMGB1、丙二醇(MDA)、超氧化物歧化醇(SOD)、平均动脉压(MAP)、心率(HR)水平的变化及并发症的发生情况。结果:通气前,两组血清IL-6、IL-10、MIP-2、SP-D、HMGB1、MDA、SOD、MAP、HR水平比较差异均无统计学意义(P0.05)。通气后,两组血清IL-6、IL-10、MIP-2、SP-D、HMGB1、MDA水平均较治疗前显著上升,而实验组以上指标均明显低于对照组;两组血清SOD、MAP、HR均较治疗前显著降低,且研究组低于对照组,差异均有统计学意义(P0.05)。结论:七氟烷吸入麻醉能够抑制单肺通气患者血清IL-6、IL-10、MIP-2、SP-D及HMGB1水平上升,改善机体的氧化应激状态,利于血流动力学的稳定,发挥肺部保护作用。     

鼻炎患者血清中IL-27、IL-17、IL-10在变应性的表达及临床意义

目的:探讨鼻炎患者血清中白细胞介素-27(IL-27)、白细胞介素-17(IL-17)、白细胞介素-10(IL-10)在变应性中的表达以及临床意义。方法:选取2013年5月到2014年5月我院收治的变应性鼻炎患者60例为研究组,另外选取健康志愿者60例为对照组,应用酶联免疫吸附试验(ELISA)检测入选者血清中的IL-27、IL-17以及IL-10的表达情况,并分析其相关关系。结果:研究组血清中IL-27、IL-10水平显著低于对照组,差异具有统计学意义(P0.05);研究组IL-17水平显著高于对照组,差异具有统计学意义(P0.05);研究组患者血清中的IL-27和IL-17呈负相关关系(r=-0.372,P=0.035),IL-27和IL-10呈正相关关系(r=0.524,P=0.026),而IL-17和IL-10无相关关系(r=0.519,P=0.318)。结论:变应性鼻炎患者中IL-27和IL-10呈低表达,而IL-17呈高表达,IL-27可能对IL-17和IL-10具有免疫调节的作用。     

甲状腺癌患者血清IL-17、IL-35、SIL-2R表达水平及其临床意义

目的:探讨甲状腺癌患者血清白细胞介素-17(IL-17)、白细胞介素-35(IL-35)及可溶性白介素-2受体(SIL-2R)水平及其对甲状腺癌诊断与病情评估的临床价值。方法:选取我院2015年6月~2016年12月收治的甲状腺腺瘤患者38例、甲状腺癌患者49例为研究对象,另选取同期于我院体检中心接受体检的52例健康体检者为对照组。采用酶联免疫吸附法(ELISA)检测和比较其血清IL-17、IL-35、SIL-2R水平,并分析甲状腺癌患者血清IL-17、IL-35、SIL-2R水平与其年龄、病程、病理分期的相关性。结果:甲状腺腺瘤组血清IL-17、IL-35、SIL-2R水平与对照组比较差异均无统计学意义(P0.05)。甲状腺癌组血清IL-35水平显著低于甲状腺腺瘤组和对照组(P0.01),血清IL-17、SIL-2R水平均显著高于甲状腺瘤组和对照组(P0.01)。血清IL-17、SIL-2R水平随甲状腺癌分化程度的降低而升高,血清IL-35水平随甲状腺癌分化程度的降低而降低(P0.01)。血清IL-17、SIL-2R水平随甲状腺癌病理分期的增加而升高,血清IL-35水平随甲状腺癌病理分期的增加而降低(P0.01)。血清IL-17、SIL-2R水平与甲状腺癌病理分期均呈显著正相关(r=0.432、0.439,P均0.05)。血清IL-35水平与甲状腺癌病理分期呈显著负相关(r=-0.602,P0.05)。血清IL-17与IL-35呈显著负相关(r=-0.323,P0.05),IL-17与SIL-2R呈显著正相关(r=0.429,P0.05),IL-35与SIL-2R呈显著负相关(r=-0.415,P0.05)。结论:甲状腺癌患者的血清IL-17、SIL-2R水平均显著上调,IL-35水平显著下调,其对甲状腺癌的早期诊断、病情评估均具有重要参考价值。     

普适泰与地奥司明对老年慢性前列腺炎患者血清和前列腺液中MIP-2和MIP-1α的影响

目的:探讨普适泰联合地奥司明治疗老年慢性前列腺炎(CP)的临床效果及对患者血清和前列腺液巨噬细胞炎性蛋白-2(MIP-2)、巨噬细胞炎性蛋白-1α(MIP-1α)水平的影响。方法:选择我院2015年1月~2016年9月收治的126例老年CP患者,采取随机数字表将其分成两组。观察组(63例)给予普适泰联合地奥司明治疗,对照组(63例)予以普适泰治疗。记录比较两组临床疗效,治疗前后血清与前列腺液的MIP-2、MIP-1α水平;评价两组用药安全性。结果:治疗12周后,观察组总有效率为93.7%,较对照组明显升高(81.0%,P0.05)。与治疗前对比,两组治疗12周后血清和前列腺液中MIP-2、MIP-1α水平均显著下降(P0.01);且观察组治疗12周后血清与前列腺液中MIP-2、MIP-1α水平均显著低于对照组同期(P0.01)。两组不良反应发生率对比差异无统计学意义(P0.05)。结论:普适泰联合地奥司明治疗老年CP/CPPS能更安全有效地改善患者的临床症状,可能与其显著降低血清和前列腺液中MIP-2、MIP-1α水平有关。     

IL-6 和IL-23在溃疡性结肠炎患者肠粘膜中的表达及意义

目的:研究溃疡性结肠炎患者炎症粘膜中IL-6及IL-23的表达及其临床意义。方法:选取2013年4月到2014年4月我院收治的溃疡性结肠炎患者60例,根据Sutherland疾病活动指数分为轻度组(11例)、中度组(19例)、重度组(14例)、缓解期组(16例),另选健康志愿者20名为对照组。分别检测各组粘膜细胞因子IL-6及IL-23的表达水平。结果:轻度组、中度组及重度组患者的IL-6和IL-23表达水平逐渐增高,显著高于缓解期组和对照组,差异有统计学意义(P<0.05);缓解期组患者IL-23水平显著高于对照组,差异有统计学意义(P<0.05);而IL-6表达与对照组比较无显著差异(P>0.05)。结论:IL-6和IL-23在溃疡性结肠炎的发生与发展中起重要作用,其表达能够反应溃疡性结肠炎的炎性程度。     

细菌性脓毒症患者中性粒细胞表面CD64及巨噬细胞炎性蛋白1α表达水平及诊断价值分析

目的 探讨细菌性脓毒症患者中性粒细胞表面CD64（nCD64）及巨噬细胞炎性蛋白1α（MIP-1α）的表达水平和诊断价值。 方法 选取2015年6月至2019年3月我院诊治的305例细菌性脓毒症患者为研究对象,根据急诊科脓毒症相关病死率评分（MEDS）结果将入选患者分为高危组和中低危组,另选取50例健康体检者作为健康对照组,检测外周血nCD64、MIP-1α表达水平,分析nCD64、MIP-1α对细菌性脓毒症的诊断价值。 结果 细菌性脓毒症患者血清nCD64、MIP-1α水平均明显高于健康对照组,高危组患者血清nCD64、MIP-1α水平明显高于中低危组,差异有统计学意义（均P结论 外周血nCD64、MIP-1α表达水平可反应细菌性脓毒症的严重程度,对细菌性脓毒症具有较高诊断价值     

趋化因子MIP-2和MCP-1在结直肠癌中表达及临床病理意义

目的:观察结直肠癌组织中趋化因子巨噬细胞炎性蛋白-2(MIP-2)和单核细胞趋化蛋白-1(MCP-1)蛋白的表达,分析其表达的临床病理意义.方法:收集结直肠癌病例的病理资料和临床资料,应用免疫组织化学SP法检测待测蛋白的表达.结果:MIP-2在癌旁肠组织组和腺癌组的阳性表达率分别为76%,68%,差异无统计学意义.MCP-1在癌旁肠组织组和腺癌组的阳性表达率分别为56%,72%,差异无统计学意义.MIP-2表达与淋巴结转移有关,而与肿块大小、部位、分化程度、浸润肠壁深度无关.MCP-1表达与肿块直径、浸润肠壁深度、淋巴结转移有关,而与部位、分化程度无关.结论:MIP-2和MCP-1在结直肠癌中的表达主要与侵袭和转移相关.     

骨髓基质细胞IL-1beta、IL-6、SCF及TPO在老年多发性骨髓瘤组织中 的表达及意义

目的:探讨老年多发性骨髓瘤患者骨髓基质细胞白细胞介素-1(IL-1β)、白细胞介素-6(IL-6)、肝细胞因子(SCF)、血小板生成素(TPO)水平及临床意义。方法:选择本院2011年1月-2014年6月收治的老年多发性骨髓瘤患者作为观察组,另选择同期参加体检的志愿者作为对照组。采集两组人员骨髓标本,制备总RNA、反转录反应、聚合酶链反应及PCR产物电泳检测等过程的测定两组患者IL-1β、IL-6、SCF、TPO水平。结果:观察组患者IL-1β、IL-6、SCF、TPO水平均明显高于对照组(P<0.05);初发患者、复发患者及移植患者的IL-1β、IL-6比较无明显差异(P>0.05)。移植组SCF水平显著低于初发组和复发组(t初移=4.967,t难移=5.169,P<0.05);初发组TPO显著高于复发组和移植组(t初难=4.736,t初移=3.568,P<0.05)。结论:老年多发性骨髓瘤患者骨髓基质细胞白细胞介素-1、白细胞介素-6、肝细胞因子、血小板生成素水平表达升高,在疾病的发生及发展过程中发挥重要作用。     

人胎脾细胞IL-2和IL-6的产生及其与NK细胞功能发育的关系

人类胚胎发育时期,脾细胞IL-2和IL-6的产生及其与NK细胞功能发育关系的研究结果表明,胚胎20周龄前IL-2的活性和NK活性细胞基本缺乏,但可分泌低水平的IL-6;随个体发育,IL-2、IL-6的产生和NK细胞活性均逐渐增强,三者间呈直线正相关关系(r>0.86);出生前,IL-6的产生和NK细胞活性显著低于成人组(p<0.01),而IL-2的产生巳达成人水平(p>0.05)。最后,对在胚胎发育过程中IL-2和IL-6的产生,及其与NK细胞功能发育间的关系进行了讨论。     

人IL-6和TNF突变体重组融合蛋白表达质粒的构建及在大肠杆菌中的表达

本文利用PCR技术,对人肿瘤坏死因子α(hTNFα)基因进行了改造,并将其与人白细胞介素-6(hIL-6)成熟肽编码区cDNA进行融合,构建了5′IL-6-TNF△融合蛋白的表达质粒pBVIL6-TNFA△。DNA序列分析证明,PCR扩增片段核苷酸序列与引物设计序列及相应的cDNA序列完全一致;重组子用限制性内切酶酶切鉴定,含有正确的IL6-TNF△融合cDNA片段;表达产物经SDS-聚丙烯酰胺凝胶电泳,分子量约为37kD,与预计的相符合;生物学活性分析初步表明,该融合蛋白具有抗肿瘤活性。     

Th2 Cytokines Augment IL-31/IL-31RA Interactions via STAT6-dependent IL-31RA Expression

Interleukin 31 receptor α (IL-31RA) is a novel Type I cytokine receptor that pairs with oncostatin M receptor to mediate IL-31 signaling. Binding of IL-31 to its receptor results in the phosphorylation and activation of STATs, MAPK, and JNK signaling pathways. IL-31 plays a pathogenic role in tissue inflammation, particularly in allergic diseases. Recent studies demonstrate IL-31RA expression and signaling in non-hematopoietic cells, but this receptor is poorly studied in immune cells. Macrophages are key immune-effector cells that play a critical role in Th2-cytokine-mediated allergic diseases. Here, we demonstrate that Th2 cytokines IL-4 and IL-13 are capable of up-regulating IL-31RA expression on both peritoneal and bone marrow-derived macrophages from mice. Our data also demonstrate that IL-4Rα-driven IL-31RA expression is STAT6 dependent in macrophages. Notably, the inflammation-associated genes Fizz1 and serum amyloid A (SAA) are significantly up-regulated in M2 macrophages stimulated with IL-31, but not in IL-4 receptor-deficient macrophages. Furthermore, the absence of Type II IL-4 receptor signaling is sufficient to attenuate the expression of IL-31RA in vivo during allergic asthma induced by soluble egg antigen, which may suggest a role for IL-31 signaling in Th2 cytokine-driven inflammation and allergic responses. Our study reveals an important counter-regulatory role between Th2 cytokine and IL-31 signaling involved in allergic diseases.     

Immunolocalization of IL-17A, IL-17B, and their receptors in chondrocytes during fracture healing.

Fracture healing in long bones is a sequential multistep cascade of hemostasis, transient inflammation, chemotaxis of progenitor cells, mitosis, differentiation of cartilage, and replacement with bone. This multistep cascade is orchestrated by cytokines and morphogens. Members of the interleukin (IL)-17 family, including IL-17B, have been identified in cartilage, but their expression during fracture healing is unknown. In this study, we determined the immunolocalization of cytokines IL-17A and IL-17B, along with the IL-17 receptor (IL-17R) and IL-17 receptor-like protein (IL-17RL), during the sequence of fracture repair in a standard model. The results were extended to developmental changes in the epiphyseal growth plate of long bones. Members of the IL-17 family were localized in chondrocytes in the fracture callus. Moreover, we found significant parallels to the localization of these cytokines and their receptors in chondrocytes during an endochondral differentiation program in the epiphyseal growth plate.     

IL-17 inhibits human Th1 differentiation through IL-12Rβ2 downregulation

Th17 cells are critical in adaptive immunity and autoimmune disease. The polarized development of Th17, Th1 and Th2 cells is dependent on counterregulatory effects on each other. Whereas IFN-γ inhibits Th17 development, the effect of IL-17 in human Th1 development is not known. We report a novel negative regulatory role of IL-17 on IL-12Rβ2 expression associated with reduced IL-12 responsiveness. IL-17 decreased IL-12-induced IFN-γ expression in PBMC and developing Th1 cells, associated with a selective reduction in IL-12Rβ2, and not IL-23R, IL-12Rβ1 or T-bet. Counterregulatory effects of human Th17 on Th1 lineage cytokines may contribute to lineage divergence. In autoimmune disease, IL-17 may reinforce its own developmental programme by reducing IL-12 responsiveness, thus limiting inhibitory effects of IFN-γ on Th17 development.     

慢病毒载体介导shRNA干扰IL-6Rα基因表达削弱IL-6对猪脂肪细胞分化的影响

为研究白细胞介素-6(IL-6)对猪脂肪细胞分化的影响及其分子机制,构建猪IL 6Rα基因RNA干扰（RNA interference, RNAi）慢病毒载体;用IL-6Rα-RNAi重组慢病毒预处理原代培养的猪前体脂肪细胞或不处理, 然后用100 ng/mL IL-6处理分化第6 d的脂肪细胞48 h．通过测定甘油释放量检测脂肪细胞的脂解率;油红O染色提取法测定脂肪细胞的脂质含量;采用RT-PCR 和Western印迹检测脂肪细胞分化相关基因的mRNA 和蛋白表达．结果显示,IL-6显著抑制猪脂肪细胞分化,并下调PPARγ2、Perilipin A和IRS-1的mRNA及蛋白表达,同时增强ERK1/2磷酸化;IL-6Rα-RNAi预处理前体脂肪细胞则显著逆转IL 6的上述作用．总之,IL-6通过多重机制抑制猪脂肪细胞分化;而且本研究构建的IL-6Rα-RNAi重组慢病毒载体可有效阻断IL-6信号,为进一步研究IL-6的功能奠定了基础．     

Context-dependent IL-6 potentiation of interferon- gamma-induced IL-12 secretion and CD40 expression in murine microglia

Interleukin-6 (IL-6) is produced by neurons, astrocytes, and microglia, and elevated levels of IL-6 within the CNS have been documented in multiple neurological disorders including Alzheimer's disease, stroke, epilepsy, attention deficit disorder, cerebral palsy, and multiple sclerosis. Here, we sought to understand how IL-6 regulates microglial signal transduction and their immune properties. Using highly enriched cultures of neonatal murine microglia we show that IL-6 alone has direct effects on microglia as it activates STAT3 and extracellular signal-regulated kinase pathways in a time- and dose-dependent fashion and it enhances interferon-gamma (IFNγ)-stimulated IL-12 secretion. However, other immune properties were only weakly modulated by IL-6 when administered without the soluble IL-6 receptor (sIL-6R). For instance, IFNγ-induced expression of the co-stimulatory molecule, CD40 was dependent on sIL-6R administration. IL-6 with or without sIL-6R did not affect major histocompatability complex class II expression. In granulocyte–macrophage colony-stimulating factor (GMCSF)-induced dendritic cell-like microglia, IL-6/sIL-6R and IFNγ stimulated an even greater increase in CD40 expression compared with primary microglia. Altogether, our results demonstrate that microglial responses to IL-6 are not simple in that the effects of IL-6 are context-dependent. In particular, the presence or absence of sIL-6R, IFNγ or GMCSF will alter the type and amplitude of their response.     

TGF-β is necessary for induction of IL-23R and Th17 differentiation by IL-6 and IL-23

TGF-β and IL-6 induce Th17 differentiation, and IL-23 is required for expansion and maintenance of Th17 cells. Recently, it was shown that IL-6 up-regulates IL-23R mRNA in naive CD4⁺ T cells and therefore IL-6 and IL-23 synergistically promote Th17 differentiation. However, the molecular mechanism whereby IL-6 and IL-23 induce Th17 differentiation and the relevance to TGF-β remain unknown. Here, we found that IL-6 up-regulated IL-23R mRNA expression, and IL-6 and IL-23 synergistically augmented its protein expression. The combination induced Th17 differentiation, and TGF-β1 further enhanced it. IL-6 augmented endogenous TGF-β1 mRNA expression, whereas the amount of TGF-β produced was not enough to induce Th17 differentiation by IL-6 alone. However, unexpectedly, the up-regulation of IL-23R and induction of Th17 differentiation by IL-6 and IL-23 were almost completely inhibited by anti-TGF-β. These results suggest that the induction of IL-23R and Th17 differentiation by IL-6 and IL-23 is mediated through endogenously produced TGF-β.     

H2O2 induces oxidative stress damage through the BMP-6/SMAD/hepcidin axis

Age-related macular degeneration (AMD) is one of the leading causes of blindness in elderly individuals worldwide. Oxidative stress injury to retinal pigment epithelial (RPE) cells plays a major role in the pathogenesis of AMD. The purpose of this study was to observe the correlation between Hepcidin and neovascular age-related macular degeneration (nAMD) and to further observe whether oxidative stress can inhibit Hepcidin expression through relevant signaling pathways to produce oxidative damage. We compared the concentrations of Hepcidin in the aqueous humor of nAMD patients and a control group and found that the concentration of Hepcidin was lower in nAMD patients. Through PCR and western blotting, we observed that H₂O₂ can significantly inhibit the expression of Bone morphogenetic protein-6 (BMP-6) and Hepcidin and increase the intracellular iron concentration in RPE cells, while BMP-6 can reverse the inhibition of Hepcidin and the increase in iron concentration caused by H₂O₂. In addition, alterations in smad1 and smad5 expression were examined, and pretreatment with BMP-6 was demonstrated to reduce H₂O₂-induced activation of smad1 and smad5. The effects of BMP-6 were attenuated by smad1 and smad5 siRNA, further verifying that oxidative stress inhibits the expression of Hepcidin by inhibiting activation of the BMP/SMAD signaling pathway. To some extent, this study verified that oxidative stress injury plays a role in nAMD by affecting the level of hepcidin, which lays a foundation for exploring new methods to treat nAMD.     

融合蛋白pIL6-IL2在E.coli中的表达及活性鉴定

白细胞介素 2 (interleukin 2 ,IL-2 )与白细胞介素 6(interleukin 6,IL-6)能分别刺激T淋巴细胞增殖与B淋巴细胞分泌免疫球蛋白 ,从而促进动物机体的细胞免疫与体液免疫 ;另外IL2与IL6在发挥生物学活性时还有相互协同作用。因此 ,将去除信号肽的猪白细胞介素 6(pIL-6)与猪白细胞介素 2 (pIL-2 )cDNAs序列通过一段Linker相连 ,克隆到E .coli表达载体pPET-2 8a中。该融合蛋白IL6-IL2在E .coli表达菌BL2 1 (DE3)中获得成功表达 ,SDS-PAGE分析分子量约为40kDa ,表达量达到总菌体的 66.26%。用IL6依赖的B9细胞与IL2依赖的CTLL细胞增殖试验进行融合蛋白IL6 IL2的生物活性检测 ,其活性可分别达到 0.8× 10³U /mg和 6.4× 10³ U/mg。