Study of tumor necrosis factor receptor in the inflammatory bowel disease

Ulcerative colitis(UC)and Crohn’s disease(CD)are part of Inflammatory Bowel Diseases(IBD)and have pathophysiological processes such as bowel necrosis and enteric neurons and enteric glial cells.In addition,the main inflammatory mediator is related to the tumor necrosis factor-alpha(TNF-α).TNF-αis a mediator of the intestinal inflammatory processes,thus being one of the main cytokines involved in the pathogenesis of IBD,however,its levels,when measured,are present in the serum of patients with IBD.In addition,TNF-αplays an important role in promoting inflammation,such as the production of interleukins(IL),for instance IL-1βand IL-6.There are two receptors for TNF as following:The tumor necrosis factor 1 receptor(TNFR1);and the tumor necrosis factor 2 receptor(TNFR2).They are involved in the pathogenesis of IBD and their receptors have been detected in IBD and their expression is correlated with disease activity.The soluble TNF form binds to the TNFR1 receptor with,and its activation results in a signaling cascade effects such as apoptosis,cell proliferation and cytokine secretion.In contrast,the transmembrane TNF form can bind both to TNFR1 and TNFR2.Recent studies have suggested that TNF-αis one of the main pro-inflammatory cytokines involved in the pathogenesis of IBD,since TNF levels are present in the serum of both patients with UC and CD.Intravenous and subcutaneous biologics targeting TNF-αhave revolutionized the treatment of IBD,thus becoming the best available agents to induce and maintain IBD remission.The application of antibodies aimed at neutralizing TNF-αin patients with IBD that induce a satisfactory clinical response in up to 60%of patients,and also induced long-term maintenance of disease remission in most patients.It has been suggested that anti-TNF-αagents inactivate the pro-inflammatory cytokine TNF-αby direct neutralization,i.e.,resulting in suppression of inflammation.However,anti-TNF-αantibodies perform more complex functions than a simple blockade.

Effects of Irbesartan and Metformin on tumor necrosis factor receptor and monocyte chemotactic protein 1 in patients with early diabetic nephropathy

Objective: To explore the effect of Irbesartan and Metformin on tumor necrosis factor receptor 1 and monocyte chemoattractant protein-1 in patients with early diabetic nephropathy. Methods: A total of 162 patients with early diabetic nephropathy who had been admitted to the Hospital between February 2017 and February 2018 were randomly assigned into a Metformin group, an Irbesartan group, and a combination therapy group. The Metformin group were treated with oral Metformin, those in the Irbesartan group were given oral Irbesartan for treatment, and the combination therapy group was treated with Metformin combined with Irbesartan. After 3 months of continuous treatment, the levels of sTNFR1, high-sensitivity C-reactive protein, monocyte chemoattractant protein-1, glucose metabolism index, proteinuria, and serum creatinine levels in the two groups were compared. Results:After treatment, the levels of sTNFR1, sICAM-1, hs-CRP, and MCP-1 in the three groups decreased compared with those before treatment, and the levels in the combination therapy group were all shown to be lower than those of the Metformin group and the Irbesartan group, with statistically significant differences (P<0.05). The levels of glycosylated hemoglobin and fasting blood glucose in the three groups were significantly lower than before treatment, and those in the combination therapy group were lower than the Metformin group and Irbesartan group, where the difference was statistically significant (P<0.05). The 24-hour urinary protein quantification, urinary albumin excretion rate, and serum creatinine in the combination therapy group were lower than those in the Metformin group and in the Irbesartan group, where the differences were statistically significant (P<0.05). Conclusion: The effects of metformin combined with irbesartan on early diabetic nephropathy patients were significant, which can effectively reduce the levels of serum sTNFR1 and MCP-1, relieve inflammation and improve glucose metabolism and proteinuria level.

Effect of Llinagliptin on tumor necrosis factor receptor and monocyte chemoattractant protein-1 in patients with diabetic nephropathy

Objective:To explore the effect of Linagliptin on tumor necrosis factor receptor and monocyte chemoattractant protein-1 in patients with diabetic nephropathy.Methods: A total of 98 patients with diabetic nephropathy admitted to the Hospital from January 2017 to September 2018 were enrolled. The patients were divided into two groups according to the random double-blind method, with 49 cases in each group. The control group was treated with Metformin, whereas the experimental group was treated with Linagliptin plus Metformin. After 3 months of continuous treatment, the renal function [urinary albumin excretion rate, 24 h urine protein quantitation and serum creatinine], glycolipids metabolic levels [glycated hemoglobin, fasting blood glucose, total cholesterol and triglycerides], monocyte chemoattractant protein-1, tumor necrosis factor receptor, high-sensitivity C-reactive protein, and adverse reactions were compared between the two groups.Results:After 3 months of treatment, the levels of UAER, 24 h Upor and Scr in the experimental group were shown to be lower than those in the control group, and the difference was statistically significant. After 3 months of treatment, the levels of HbA1c, FPG, TC and TG in the experimental group were shown to be lower than the control group, and the difference was statistically significant. After 3 months of treatment, the levels of MCP-1, sTNFR1 and hs-CRP in the experimental group were lower than those in the control group, and the difference was statistically significant. There was no significant difference in incidence of adverse reactions between the two groups.Conclusion: For patients with diabetic nephropathy, Linagliptin is with higher safety, which can help improve their glycolipids metabolic levels and renal function, reduce the inflammatory response and the levels of MCP-1 and sTNFR1, and yet incur fewer adverse reactions.

Interleukin-1β,Tumor Necrosis Factor-α and Lipopolysaccharide Induce Expression of Monocyte Chemoattractant Protein-1 in Calf Aortic Smooth Muscle Cells

Summary: To investigate whether interleukin-1β(IL-1β), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) induce expression of monocyte chemoattractant protein-1 (MCP-1 ) mRNA and protein in calf aortic smooth muscle cells(SMCs), calf aortic SMCs were cultured by a substrate-attached explant method. The cultured SMCs were used between the third to the fifth passage. After the cells became confluent, the SMCs were exposed to 2 ng/ml IL-1β, 20ng/ml TNF-1α and 100 ng/ml LPS respectively, and the total RNA of SMCs which were incubated for 4 h at 37℃ were extracted from the cells by using guanidinium isothiocyanate method. The expres- ion of MCP-1 mRNA in SMCs was detected by using dot blotting analysis using a probe of γ-32 P- end-labelled 35-mer oligonucleotide. After a 24-h incubation, the media conditioned by the cul- tured SMCs were collected. The MCP-1 protein content in the conditioned media was determined by using sandwich ELISA. The results were as follows: Dot blotting analysis showed that the cul- tured SMCs could express MCP-1 mRNA. After a 4-h exposure to IL-1β, TNF-α and LPS, the MCP-1 mRNA expression in SMCs was increased (3.6-fold, 2. 3-fold and 1. 6-fold, respectively). ELISA showed that the levels of MCP-1 protein in the conditioned media were also increased (2.9- fold, 1.7-fold and 1.1-fold, respectively). The results suggest that calf aortic SMCs could ex- press MCP-1 mRNA and protein. IL-1β and TNF-α can induce strong expression of MCP-1 mRNA and protein, and the former is more effective than the latter.

Increased tumor necrosis factor receptor 1 expression in human colorectal adenomas

AIM: To determine the expression statuses of tumor necrosis factor (TNF)-α, its receptors (TNF-R) and downstream effector molecules in human colorectal adenomas. METHODS: We measured the serum concentrations of TNF-α and its receptors in 62 colorectal adenoma patients and 34 healthy controls. The protein expression of TNF-α, TNF-R1, TNF-R2 and downstream signals of the TNF receptors, such as c-Jun N-terminal kinase (JNK), nuclear factor-κ B and caspase-3, were also investigated in human colorectal adenomas and in normal colorectal mucosal tissues by immunohistochemistry. Immunofluorescence confocal microscopy was used to investigate the consistency of expression of TNF-R1 and phospho-JNK (p-JNK). RESULTS: The serum levels of soluble TNF-R1 (sTNF-R1) in adenoma patients were significantly higher than in the control group (3.67 ± 0.86 ng/mL vs 1.57 ± 0.72 ng/mL, P < 0.001). Receiver operating characteristic analysis revealed the high diagnostic sensitivity of TNF-R1 measurements (AUC was 0.928) for the diagnosis of adenoma, and the best cut-off level of TNF-R1 was 2.08 ng/mL, with a sensitivity of 93.4% and a specificity of 82.4%. There were no significant differences in the serum levels of TNF-α or sTNF-R2 between the two groups. Immunohistochemistry showed high levels of TNF-R1 and p-JNK expression in the epithelial cells of adenomas. Furthermore, a high incidence of co-localization of TNF-R1 and p-JNK was identified in adenoma tissue. CONCLUSION: TNF-R1 may be a promising biomarker of colorectal adenoma, and it may also play an important role in the very early stages of colorectal carcinogenesis.

Tumor necrosis family receptor superfamily member 9/tumor necrosis factor receptor-associated f

Hepatitis C virus(HCV)infection is an excellent immunological model for understanding the mechanisms developed by non-cytopathic viruses and tumors to evade the adaptative immune response.The antigen-specific cytotoxic T cell response is essential for keeping HCV under control,but during persistent infection,these cells become exhausted or even deleted.The exhaustion process is progressive and depends on the infection duration and level of antigenemia.During high antigenic load and long duration of infection,T cells become extremely exhausted and ultimately disappear due to apoptosis.The development of exhaustion involves the impairment of positive co-stimulation induced by regulatory cytokines,such as transforming growth factor beta 1.This cytokine downregulates tumor necrosis factor receptor(TNFR)-associated factor 1(TRAF1),the signal transducer of the T cell co-stimulatory molecule TNFR superfamily member 9(known as 4-1BB).This impairment correlates with the low reactivity of T cells and an exhaustion phenotype.Treatment with interleukin-7 in vitro restores TRAF1 expression and rescues T cell effector function.The process of TRAF1 loss and its in vitro recovery is hierarchical,and more affected by severe disease progression.In conclusion,TRAF1 dynamics on T cells define a new pathogenic model that describes some aspects of the natural history of HCV,and sheds light on novel immunotherapy strategies for chronic viral infections and cancer.

Tumor necrosis factor alpha receptor 1 deficiency in hepatocytes does not protect from non-alcoholic steatohepatitis, but attenuates insulin resistance in mice

BACKGROUND End-stage liver disease caused by non-alcoholic steatohepatitis(NASH)is the second leading indication for liver transplantation.To date,only moderately effective pharmacotherapies exist to treat NASH.Understanding the pathogenesis of NASH is therefore crucial for the development of new therapies.The inflammatory cytokine tumor necrosis factor alpha(TNF-α)is important for the progression of liver disease.TNF signaling via TNF receptor 1(TNFR1)has been hypothesized to be important for the development of NASH and hepatocellular carcinoma in whole-body knockout animal models.AIM To investigate the role of TNFR1 signaling in hepatocytes for steatohepatitis development in a mouse model of diet-induced NASH.METHODS NASH was induced by a western-style fast-food diet in mice deficient for TNFR1 in hepatocytes(TNFR1ΔHEP)and their wild-type littermates(TNFR1fl/fl).Glucose tolerance was assessed after 18 wk and insulin resistance after 19 wk of feeding.After 20 wk mice were assessed for features of NASH and the metabolic syndrome such as liver weight,liver steatosis,liver fibrosis and markers of liver inflammation.RESULTS Obesity,liver injury,inflammation,steatosis and fibrosis was not different between TNFR1ΔHEP and TNFR1fl/fl mice.However,Tnfr1 deficiency in hepatocytes protected against glucose intolerance and insulin resistance.CONCLUSION Our results indicate that deficiency of TNFR1 signaling in hepatocytes does not protect from diet-induced NASH.However,improved insulin resistance in this model strengthens the role of the liver in glucose homeostasis.

The interleukin-1 receptor antagonist (IL-1-Ra) and soluble tumor necrosis factor receptor I (sTNF RI) in periodontal disease

The course and severity of periodontitis can be significantly affected by bacterial virulence as well as host immunity dysfunction. Periodontal tissue destruction has been proved to result from cascade of cytokines synthesized by reactive cells upon stimulation by pathogenic bacteria and lipopolysaccharides within their cell membranes. The clinical use of genetically programmed cells, producing substances blocking IL-1, based on recombinant IL-1 antagonist, as well as cytokines activating fibroblasts and osteoblasts to regenerate the destroyed periodontal tissue could prove alternative to the conventional treatment. Another cytokine of interest in respect to periodontitis ethiopathogenesis is soluble tumor necrosis factor receptor I (sTNF RI). Observation of soluble TNF receptors as physiologic inhibitors of TNF led to its administration in therapeutic process as well as in therapy selected cases of aggressive periodontitis.

Increase of tumor necrosis factor receptor 1 expression in women with unexplained early spontaneous abortion

To investigate membrane tumor necrosis factor receptor 1 protein expression level in decidua andconcentration of soluble tumor necrosis factor receptor 1 in serum in women with unexplained early spontaneous abortion,threatened abortion, and compare the levels with healthy pregnant women. Methods: Thirty-seven women with unexplainedearly spontaneous abortion, 27 women with threatened abortion, and 34 healthy pregnant women undergoing artificial abortionof pregnancy at 6 - 10 weeks of gestation were selected. Decidual samples were collected when women were undergoing arti-ficial abortion, and blood samples were collected at the same time. The level of membrane tumor necrosis factor receptor 1 indecidua was detected by flow cytometer, and the concentration of soluble tumor necrosis factor receptor 1 in sera was mea-sured with an enzyme-linked immunosorbent assay. Results: The ercentages of membrane tumor necrosis factor receptor 1positive decidual cells were 16.42 ± 7.10 Mean ± SD for women with unexplained early spontaneous abortion and 13.14 ±6.30 for healthy pregnant women ( P ＜ 0.05). Serum oncentration of soluble tumor necrosis factor receptor 1 was signifi-cantly higher in women with unexplained early spontaneous abortion than in healthy pregnant women and in women withthreatened abortion, and no difference was found between healthy pregnant women and women with threatened abortion.Conclusion: Women with unexplained early spontaneous abortion present significantly higher expression of tumor necrosisfactor receptor 1 than healthy pregnant women, suggesting that over-expression of tumor necrosis factor receptor 1 may cont-ribute to the development of early spontaneous abortion.

血清Ephrin-A1、CTRP9对糖尿病性视网膜病变的诊断价值及与机体氧化应激的相关性分析

目的分析血清肝配蛋白A1(Ephrin-A1)、C1q肿瘤坏死因子相关蛋白9(CTRP9)对糖尿病性视网膜病变的诊断价值及与机体氧化应激的相关性。方法回顾性选择自2021年1月至2023年1月新疆维吾尔自治区人民医院接诊的80例糖尿病性视网膜病变患者作为观察组,另选同期的80例单纯2型糖尿病患者作为对照组。检测两组患者血清Ephrin-A1、CTRP9及氧化应激指标[超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、丙二醛],分析不同分期的糖尿病性视网膜病变患者血清Ephrin-A1、CTRP9水平的差异性,使用受试者工作特征(ROC)曲线分析血清Ephrin-A1、CTRP9对糖尿病性视网膜病变的诊断效能,Pearson相关性分析糖尿病性视网膜病变患者血清Ephrin-A1、CTRP9水平与机体氧化应激指标的相关性。结果观察组血清Ephrin-A1水平为(7.81±2.34)ng/mL,高于对照组[(2.25±0.76)ng/mL],CTRP9水平为(98.17±10.13)pg/mL,低于对照组[(156.42±15.89)pg/mL],差异均有统计学意义(P<0.05)。观察组血清SOD、GSH水平分别为(50.14±5.63)U/L、(142.34±13.98)mg/L,均低于对照组[(73.52±8.52)U/L、(189.71±23.56)mg/L],丙二醛水平为(6.89±3.07)μmol/L,高于对照组[(3.56±1.02)μmol/L],差异均有统计学意义(P<0.05)。增生型糖尿病性视网膜病变患者血清Ephrin-A1为(15.42±4.80)ng/mL,高于非增生型糖尿病性视网膜病变患者[(6.09±2.11)ng/mL],CTRP9水平为(75.25±6.73)pg/mL,低于非增生型糖尿病性视网膜病变患者[(119.46±13.08)pg/mL],差异均有统计学意义(P<0.05)。经ROC曲线分析,血清Ephrin-A1联合CTRP9诊断糖尿病性视网膜病变的敏感度为92.68%、特异度为53.69%、AUC为0.931。经Pearson相关性分析,糖尿病性视网膜病变患者血清Ephrin-A1与SOD、GSH呈负相关,与丙二醛呈正相关(P<0.05);CTRP9水平与SOD、GSH呈正相关,与丙二醛呈负相关(P<0.05)。结论血清Ephrin-A1联合CTRP9可提高对糖尿病性视网膜病变的诊断效能,其中机体氧化应激与Ephrin-A1呈正性关联,与CTRP9呈负性关联,值得进一步研究应用。

miR-126-5p通过靶向TRAF3抑制糖氧剥夺再灌注介导的HT22细胞凋亡和炎症

目的探讨miR-126-5p通过靶向肿瘤坏死因子受体相关因子3(tumor necrosis factor receptor-associated factor 3,TRAF3)对糖氧剥夺再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)介导的小鼠海马神经元细胞HT22细胞凋亡和炎症的影响。方法模拟缺血/再灌注损伤(ischemia/reperfusion,I/R)损伤在体外建立氧糖剥夺/复氧(oxygen-glucose deprivation/reperfusion,OGD/R)细胞模型,分析miR-126-5p与TRAF3靶向关系及对HT22细胞凋亡和炎症反应的影响。结果与对照组比较,OGD/R组中miR-126-5p下调而TRAF3 mRNA及蛋白水平上调,细胞存活率及Bcl-2蛋白水平降低,乳酸脱氢酶(lactate dehydrogenase,LDH)释放量、细胞凋亡率、Bax及Cleaved caspase-3蛋白水平升高(P均<0.05)。与OGD/R+mimic-NC组比较,OGD/R+miR-mimic组、OGD+miR-mimic+pcDNA组TRAF3蛋白水平、LDH释放量、细胞凋亡率、Bax及Cleaved caspase-3蛋白水平明显降低,细胞存活率及Bcl-2蛋白水平升高,而OGD+miR-mimic+pcDNA-TRAF3组各指标升高,细胞存活率明显下降(P均<0.05)。结论miR-126-5p通过靶向TRAF3,抑制OGD/R介导的HT22细胞凋亡和炎症反应,从而对神经元细胞发挥保护作用。

补体因子H相关蛋白1促进巨噬细胞分泌肿瘤坏死因子-α调控足细胞增殖和迁移实验研究

目的:探讨补体因子H相关蛋白1(CFHR1)通过巨噬细胞分泌肿瘤坏死因子-α(TNF-α)调控足细胞增殖和迁移的作用。方法:体外培养人单核巨噬细胞和人肾足细胞。巨噬细胞分为对照组和CFHR1干预组,分别进行牛血清白蛋白或CFHR1重组蛋白干预24 h,ELISA法测定上清液TNF-α水平。足细胞分为空白组、TNF-α干预组、对照上清液干预组、CFHR1上清液干预组、CFHR1上清液+TNF-α中和抗体干预组。CCK8法检测各组细胞增殖。Transwell法检测各组细胞迁移。Wb法检测各组细胞中相关蛋白变化。结果:巨噬细胞的CFHR1干预组上清液中TNF-α含量显著增加(P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的细胞增殖比率和迁移数量均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的细胞增殖比率和迁移数量均显著提高(均P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的足细胞裂孔膜蛋白(Nephrin)、足突蛋白(Podocin)、纤维状肌动蛋白(F-Actin)、整合素α3β1蛋白(α3β1)表达均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的Nephrin、Podocin、F-actin、α3β1蛋白表达均显著增多(均P<0.05)。结论:CFHR1促进巨噬细胞分泌的TNF-α可显著抑制足细胞增殖水平和迁移能力,这可能是高浓度CFHR1促进肾病综合征发展的途径。

PD1/PD-L1、中性粒细胞弹性蛋白酶、赖氨酰氧化酶在膀胱癌中的表达及其与膀胱癌预后的关系

目的探讨膀胱癌组织中程序性细胞死亡受体1(PD1)、程序性细胞死亡配体1(PD-L1)、中性粒细胞弹性蛋白酶(NE)、赖氨酰氧化酶(LOX)的表达水平及其表达差异与膀胱癌患者临床病理特征和预后的关系。方法选取2018年1月至2020年12月江西省人民医院收治的56例膀胱癌患者的癌组织作为研究对象,并记录患者的复发和死亡情况进行分析研究。采用免疫组织化学染色法检测PD1、PD-L1、NE、LOX的表达情况,分析PD1、PD-L1、NE、LOX与患者的病理特征和预后的关系。结果免疫组化结果显示PD1在肿瘤相关免疫细胞中存在表达(46/56);TNM分期T2-T3期、高级别肿瘤和出现淋巴结转移的患者PD1和PDL-1的表达阳性率均明显升高,差异有统计学意义(P<0.05)。NE与PD1、PDL-1的表达呈正相关(P<0.05);LOX与PD-L1、PDL-1的表达呈正相关(P<0.05)。PD1阳性表达患者3年无进展生存率为58.70%(27/46),PD-L1阳性表达患者3年无进展生存率为43.48%(10/23),不同PD1、PD-L1表达患者的3年无进展生存率比较差异有统计学意义(P<0.05)。结论膀胱癌组织中PD1、PD-L1表达增加可作为膀胱癌患者潜在的预后预测指标,有助于筛选术后复发的高危患者,并为膀胱癌的免疫学治疗提供新靶点。

自发性急性脑出血患者血浆sCD163/sTWEAK比值与预后的关系

目的探究自发性急性脑出血(ACH)患者血浆可溶性CD163(sCD163)/可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)比值与预后的关系。方法纳入ACH患者90例作为病例组,根据格拉斯哥预后评分将病例组分为预后不良组(38例)和预后良好组(52例);另选取同期体检健康者45例为对照组。酶联免疫吸附试验检测血浆sCD163、sTWEAK水平并计算sCD163/sTWEAK比值。分析血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值与临床资料的相关性;Logistic回归分析ACH患者预后不良的影响因素;受试者工作特征(ROC)曲线分析sCD163/sTWEAK比值对ACH患者预后不良的预测价值。结果病例组血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值均显著高于对照组;预后良好组上述指标均低于预后不良组(P<0.05)。预后良好组血肿体积、美国国立卫生研究院卒中量表(NIHSS)评分、高血压及幕下出血比例均低于预后不良组,低密度脂蛋白胆固醇(LDL-C)高于预后不良组(P<0.05)。相关性分析表明,血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值与出血部位、血肿体积、NIHSS评分、白细胞计数、血小板计数、中性粒细胞/淋巴细胞比值(NLR)呈正相关(P<0.05)。Logistic回归分析显示,sCD163/sTWEAK比值、出血部位、血肿体积、NIHSS评分为ACH患者预后不良的影响因素(P<0.05)。ROC曲线结果表明,sCD163/sTWEAK比值评估ACH患者预后不良的AUC为0.850,敏感度和特异度分别为86.84%和69.23%。结论sCD163/sTWEAK比值在ACH患者血浆中水平较高,并与预后不良有关,该值对此类患者的预后有一定预测价值。

AR、SKP2、SOX10、PD-L1及TIL表达在三阴性乳腺癌中的意义

目的:探索雄激素受体(androgen receptor,AR)、S期激酶相关蛋白2(S-phase kinase-associated protein 2,SKP2)、性别决定区Y相关的HMG盒含因子10(sry-related HMG box-containing factor 10,SOX10)、程序性死亡配体1(programmed death-ligand 1,PD-L1)及肿瘤浸润性淋巴细胞(tumor infiltrating lymphocyte,TIL)在三阴性乳腺癌(triple negative breast cancer,TNBC)表达与临床病理特征和预后的关系。方法:根据苏木精-伊红染色(hematoxylineosin, HE)染色切片评判109例TNBC瘤巢内TIL的比例,采用Leica Bond-Max全自动免疫组化仪检测TNBC组织中AR、SKP2、SOX10、PD-L1的表达。分析以上各生物指标与临床病理特征间的关系,并采用kaplan-Meier、Log-rank进行生存分析。结果:95例患者获得随访,中位随访时间为48个月,中位无病生存时间(disease-free survival, DFS)为42个月,中位总生存时间(overall survival, OS)48个月。在TNBC中,AR阳性表达与淋巴结转移阴性(P=0.009)、肿瘤最大径<2 cm(P=0.008)相关,TIL高表达与低级别TNBC相关(P=0.007),SKP2阳性表达与神经/脉管侵犯阳性(P=0.011)、高级别TNBC相关(P=0.002),SOX10阳性表达与淋巴结转移阳性(P=0.022)、高级别TNBC(P=0.005)相关,PD-L1阳性表达与淋巴结转移阳性(P=0.020)、神经/脉管侵犯阳性(P=0.006)、高级别TNBC(P=0.042)相关。生存分析显示,SKP2、SOX10阳性表达与更差的DFS(P=0.007、P<0.001)和OS(P=0.013、P<0.001)相关,TIL高表达与更好的DFS(P=0.016)及OS(P=0.004)相关。在生物表志物的联合表达中,AR+/SKP2-、AR+/SOX10-与更好的DFS(P=0.004、P<0.001)及OS(P=0.007、P=0.001)相关,SOX10+/低TIL、PD-L1+/低TIL与更差的DFS(P<0.001、P=0.008)及OS(P=0.001、P=0.002)相关,AR-/低TIL者具有更差的OS(P=0.014)。SKP2(HR=4.143,95%CI为1.578~10.875)、SOX10(HR=7.578,95%CI为2.067~27.782)的阳性表达是影响TNBC患者DFS的独立预后因子,SKP2(HR=3.758,95%CI为1.400~10.084)、SOX10(HR=5.131,95%CI为1.316~20.000)及TIL(HR=0.375,95%CI为0.154~0.917)的阳性表达是TNBC患者OS的独立预后因子(P均<0.05)。结论:在TNBC中,AR阳性、TIL高表达与具有更好预后的临床病理特征相关,SKP2、SOX10和PD-L1与具侵袭性的临床病理特征相关。SKP2、SOX10及TIL表达与TNBC预后相关,提示这些生物指标可能成为TNBC新的预后因子,同时它们也有可能成为潜在的治疗靶点。

TNIP1基因单核苷酸多态性及其mRNA表达水平与老年慢性心力衰竭患者肺部感染的相关性分析

目的探讨肿瘤坏死因子诱导蛋白3相互作用蛋白1(TNFAIP3-interacting protein 1,TNIP1)基因单核苷酸多态性及其mRNA表达水平与老年慢性心力衰竭患者肺部感染的相关性。方法选择2019年10月至2022年10月于上海建工医院重症医学科就诊的130例老年慢性心力衰竭患者作为研究对象,根据是否于院内发生肺部感染分为感染组(32例)和未感染组(98例)。对TNIP1基因的两个SNP位点rs6889239(T>C)、rs17728338(A>G)进行基因分型,并检测外周血TNIP1基因的mRNA表达水平。结果TNIP1基因rs6889239位点在感染组和非感染组之间的基因型分布以及等位基因频率的差异均无统计学意义(P>0.05);两组的rs17728338位点AA、AG、GG基因型分布比较差异有统计学意义(P<0.05),且感染组等位基因G的频率显著高于未感染组(P<0.05)。相较于未感染组,感染组患者的外周血TNIP1基因mRNA表达水平显著增加,差异有统计学意义(P<0.001)。受试者工作特征(Receiver operating characteristic,ROC)曲线分析结果显示外周血TNIP1基因表达水平预测慢性心力衰竭患者发生肺部感染的灵敏度和特异度分别为71.9%和95.9%。感染组和非感染组TNIP1基因rs6889239位点不同基因型患者的外周血TNIP1基因的表达水平比较差异均无统计学意义(P>0.05),而rs17728338位点不同基因型患者的外周血TNIP1基因表达水平比较差异有统计学意义(P<0.05)。结论TNIP1基因rs17728338表达水平与老年慢性心力衰竭患者发生肺部感染有关。

抵当汤及其拆方对深静脉血栓形成大鼠高速泳动族蛋白B1影响的研究

目的:观察抵当汤及其拆方对DVT大鼠高速泳动族蛋白B1(HMGB1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的影响。方法:将150只Wistar雄性大鼠随机分为5组,抵当汤组、大黄桃仁组、水蛭地龙组、模型组、假手术组,每组30只。采用Reyers法建立下腔静脉结扎模型,术后第1、3、7天灌胃2 h后,腹腔麻醉开腹,采用酶联免疫吸附试验(ELISA)、蛋白质免疫印迹法检测高速泳动族蛋白B1(HMGB1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平,苏木精-伊红(HE)染色观察大鼠下腔静脉的病理变化,免疫荧光(IF)、实时逆转录PCR(Realtime RT-PCR)检测大鼠下腔静脉中的HMGB1表达。结果:模型组下腔静脉内皮细胞损伤严重并大量脱落,细胞肿胀明显,形成血栓,内膜形态不规则、管壁组织肿胀,胶原纤维增生,伴有炎症细胞浸润,抵当汤组、大黄桃仁组、水蛭地龙组等用药组内皮细胞不同程度损伤和炎症细胞浸润;模型组HMGB1荧光表达较假手术组明显增多,其余用药组HMGB1荧光表达不同程度减少;与假手术组比较,模型组大鼠的高速泳动族蛋白B1(HMGB1)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)血清水平、高速泳动族蛋白B1(HMGB1)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、蛋白表达、mRNA表达明显升高(P<0.01)。与模型组比较,抵当汤组、大黄桃仁组、水蛭地龙组等用药组HMGB1、IL-6、TNF-α血清水平、蛋白表达、mRNA表达明显降低(P<0.05,P<0.01)。抵当汤组、大黄桃仁组、水蛭地龙组等用药组比较,HMGB1、IL-6、TNF-α血清水平、高速泳动族蛋白B1(HMGB1)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、蛋白表达、mRNA表达比较差异有统计学意义(P<0.05,P<0.01),术后第1、3、7天比较,HMGB1、IL-6、TNF-α血清水平、蛋白表达、mRNA表达比较差异有统计学意义(P<0.05,P<0.01)。结论:抵当汤及其拆方通过降低DVT大鼠的HMGB1、IL-6、TNF-α表达,发挥对DVT的治疗作用。

肺癌组织中ERO1L、TNFRSF4的表达与患者免疫功能、炎症反应因子及预后的关系

目的探究肺癌组织中内质网氧化物蛋白(ERO1L)、肿瘤坏死因子受体4(TNFRSF4)的表达与肺癌患者免疫功能、炎症反应因子及其预后的关系。方法选取2018年7月~2020年7月于本院进行手术治疗的108例肺癌患者,收集术中留取的癌组织和癌旁组织标本。采用qRT-PCR检测ERO1L和TNFRSF4的mRNA相对表达量;使用免疫组织化学法检测ERO1L和TNFRSF4蛋白表达情况,分析二者表达水平与患者临床病理特征的关系,采用Kaplan-Meier法分析ERO1L、TNFRSF4蛋白表达水平与患者预后的关系。肺癌患者预后生存率的影响因素采用Cox多因素分析。结果肺癌患者癌组织中ERO1L mRNA表达水平显著高于癌旁组织,TNFRSF4 mRNA表达水平显著低于癌旁组织(P<0.05);肺癌组织中ERO1L蛋白高表达率显著高于癌旁组织,TNFRSF4蛋白高表达率显著低于癌旁组织(P<0.05)。ERO1L蛋白高表达组患者CD3^(+)、CD4^(+)显著低于低表达组(P<0.05),IL-1β、IL-6、TNF-α显著高于低表达组(P<0.05);TNFRSF4蛋白高表达组患者CD3^(+)、CD4^(+)显著高于低表达组,IL-1β、IL-6、TNF-α显著低于低表达组(P<0.05)。ERO1L高表达组患者3年累积生存率显著低于低表达组(Log rankχ^(2)=6.100,P=0.014),TNFRSF4高表达组患者3年累积生存率显著高于低表达组(Log rankχ^(2)=11.296,P=0.001)。肺癌组织的低分化、淋巴结转移、TNM分期为Ⅲ-Ⅳ期、ERO1L高表达、TNFRSF4低表达是影响患者生存率的危险因素。结论肺癌组织中ERO1L、TNFRSF4表达与患者免疫功能、炎症因子以及预后具有一定关系。

Effects of erythropoietin on the expression of tumor necrosis factor-alpha and Bax after facial nerve axotomy in rats

This study sought to evaluate the effect of high-dose erythropoietin （EPO; 5 000 IU/kg） on the expression of tumor necrosis factor-alpha （TNF-α） and Bax in the facial nucleus after facial nerve transection in rats. A total of 42 Wistar rats of both genders were used in this study, and 40 rats were randomly divided into 2 groups： EPO group and model group. The EPO group was treated with EPO once a day for 5 days at a dose of 5 000 IU/kg body weight. The model group was treated with saline of the same amount. At day 3 after EPO （or saline） treatment, the right facial nerves of the 40 rats were transected at the level of the stylomastoid foramen, with the left sides untreated. The remaining 2 rats that did not undergo axotomy served as the control group. The surviving motor neurons in operated rats were counted in coronal paraffin sections of the facial nucleus. The expression of TNF-a and Bax in the facial nucleus was detected by immunohistochemical staining at days 3, 7, 14, 21, and 28 after axotomy. At days 14, 21, and 28 after facial nerve axotomy, a significantly greater proportion of facial motor neurons survived in the EPO group than in the model group. After axotomy, the expression of TNF-a and Bax increased in motor neurons in both the EPO and the model groups. TNF-o expression reached its peak level at day 14 after axotomy, while Bax expression reached its peak level at day 21. TNF-α expression was much lower in the EPO group than in the model group at all time points. No significant difference in Bax expression was found between the EPO and the model groups. These results indicate that high-dose EPO treatment attenuates the increase in TNF-α expression in the facial nucleus and reduces the loss of motor neurons after facial nerve transection in rats. However, high-dose EPO treatment has little effect on Bax expression.

感染性休克患者IRAK1和TRAF6的表达变化及临床意义研究

目的探讨感染性休克患者白细胞介素-1受体相关激酶1(IRAK1)、肿瘤坏死因子相关受体6(TRAF6)的表达变化及临床意义。方法以2020年11月至2022年11月该院收治的142例感染性休克患者(感染性休克组)为研究对象,并以同期来该院进行体检的体检者为对照组。根据感染性休克组患者住院观察治疗28 d后的生存状况分为生存组100例和死亡组42例,监测感染性休克患者入院时及治疗2、4、6 d后的IRAK1、TRAF6表达变化,并记录患者急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分和序贯器官功能衰竭评估(SOFA)评分动态变化;Spearman相关性分析评价感染性休克患者IRAK1、TRAF6与APACHEⅡ评分、SOFA评分的相关性;Pearson相关性分析IRAK1与TRAF6的相关性;Logistic回归分析感染性休克患者生存状况的影响因素。通过受试者工作特征曲线分析IRAK1、TRAF6对感染性休克患者生存状况的诊断价值。结果入院时感染性休克组IRAK1、TRAF6相对表达水平显著低于对照组,APACHEⅡ评分、SOFA评分显著高于对照组,差异有统计学意义(P<0.05)。与入院时比较,治疗2、4、6 d后两组IRAK1、TRAF6相对表达水平均显著升高,APACHEⅡ评分、SOFA评分均显著降低,差异有统计学意义(P<0.05);与死亡组比较,生存组在各个相应时间点IRAK1、TRAF6相对表达水平均较高,APACHEⅡ评分、SOFA评分均较低,差异有统计学意义(P<0.05)。相关性分析显示,感染性休克患者IRAK1、TRAF6与APACHEⅡ评分、SOFA评分均呈负相关,IRAK1与TRAF6呈正相关(r=0.688,P<0.05)。IRAK1、TRAF6及APACHEⅡ评分是影响感染性休克患者生存状况的独立危险因素(P<0.05)。IRAK1、TRAF6联合诊断的曲线下面积(AUC)显著大于IRAK1单独诊断的AUC(Z=2.044,P=0.041),以及TRAF6单独诊断的AUC(Z=2.442,P=0.015)。结论感染性休克患者IRAK1、TRAF6的表达可评估患者生存及预后状况。