Brain edema and tumor necrosis factor-like weak inducer of apoptosis in rats with cerebral ischemia

BACKGROUND： Recent studies have demonstrated that tumor necrosis factor-like weak inducer of apoptosis （TWEAK） participates in brain edema. However, it is unclear whether blood-brain barrier （BBB） disruption is associated with TWEAK during the process of brain edema OBJECTIVE： To investigate the effects of TWEAK on BBB permeability in brain edema. DESIGN, TIME AND SETTING： An immunohistochemical observation, randomized, controlled animal experiment was performed at the Laboratory of Neurosurgical Anatomy, Xiangya Medical College, Central South University ＆ Central Laboratory, Third Xiangya Hospital, Central South University between January 2006 and December 2007. MATERIALS： A total of 48 adult Wistar rats were randomly divided into three groups： normal control （n = 8）, sham-operated （n = 8）, and ischemia/reperfusion （n = 32）. Rats from the ischemia/reperfusion group were randomly assigned to four subgroups according to different time points, i.e., 2 hours of ischemia followed by 6 hours （n = 8）, 12 hours （n = 8）, 1 day （n = 8）, or 12 days （n = 8） of reperfusion. METHODS： Focal cerebral ischemia/reperfusion injury was induced by middle cerebral artery occlusion （MCAO） using the suture method in rats from the ischemia/reperfusion group. Thread was introduced at a depth of 17-19 mm. Rats in the sham-operated group were subjected to experimental procedures similar to the ischemia/reperfusion group; however, the introducing depth of thread was 10 mm. The normal control group was not given any intervention. MAIN OUTCOME MEASURES： TWEAK expression was examined by immunohistochemistry; brain water content on the ischemic side was calculated as the ratio of dry to wet tissue weight; BBB permeability was measured by Evans blue extravasation. RESULTS： A total of eight rats died prior to and after surgery and an additional eight rats were randomly entered into the study. Thus 48 rats were included in the final analysis. In the ischemia/reperfusion group, TWEAK-positive cells were present in the ischemic penumbra surrounding the lamellar necrotic region in the fight cerebral hemisphere at 6 hours reperfusion and increased thereafter; by 2 days reperfusion they had reached a peak level, which was significantly higher than the sham-operated and normal control groups （P 〈 0.05）. At 6 hours reperfusion, both brain water content and Evans blue extravasation showed the same tendency for change as TWEAK expression. Pearson correlation analysis results revealed that the degree of TWEAK expression was positively correlated with brain water content （r = 0.892, P 〈 0.05）. CONCLUSION： The present results confirmed that TWEAK was involved in BBB disruption and participated in brain edema following cerebral ischemia.

Autophagy plays a protective role in advanced glycation end products-induced apoptosis of chondrocytes via regulation of tumor necrosis factor-α,nuclear factor-κ B and reactive oxygen species

Objective: To study the adverse effects of advanced glycation end products(AGEs) on chondrocytes and the role of autophagy in this process. Methods: Chondrocytes were harvested from the human articular cartilage tissues in surgery. AGEs were administered during chondrocytes culture. The rapamycin was used to induce autophagy. The cell viability was determined by 3-[4,5-dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay.The expression of tumor necrosis factor-α(TNF-α) and nuclear factor-κ B(NF-κ B) was detected by quantitative real-time polymerase chain reaction. The reactive oxygen species(ROS) production and apoptosis of the chondrocytes were determined by fluorescent probe and flow cytometer, respectively. Results: The chondrocytes viability was significantly reduced after 12 h incubation with AGEs(P<0.01)). In contrast, rapamycin pretreatment increased the chondrocytes viability through autophagy. AGEs increased TNF-α and NF-κ B mRNA expression of chondrocytes and autophagy receded or proceeded the change. AGEs increased intracellular ROS accumulation and autophagy reversed the change. AGEs accelerated chondrocytes apoptosis and autophagy suspended apoptosis. Conclusions: Accumulation of AGEs may have an adverse role for chondrocytes by increasing TNF-α and NF-κB expression, ROS accumulation and apoptosis; meanwhile, autophagy ameliorates the AGEsinduced adverse effects.

Antitumor effect of tumor necrosis factor-related apoptosis inducing ligand combined with mevastatin on a human glioma cell line SWO-38

BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor necrosis factor-related apoptosis inducing ligand （TRAIL）. OBJECTIVE： To explore the effect of TRAIL combined with mevastatin on the proliferation and apoptotic cell death of a human glioma cell line SWO-38, and to study its mechanism of action. DESIGN, TIME AND SETTING： An in vitro control experiment was performed at the Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University, between January and April 2009. MATERIALS： The human SWO-38 cell line was provided by Cell Research, Department of Animal Experimental Center of Sun Yat-sen University; human recombinant soluble TRAIL by R＆D, USA; and mevastatin by Sigma, USA. METHODS： SWO-38 cells were separately incubated in TRAIL （100, 200, 300, 400, and 500 tJg/L） and mevastatin （5, 10, 20, 30, and 40 pmol/L） for 72 hours. In addition, SWO-38 cells were incubated in TRAIL （300 μg/L）, mevastatin （30 μmol/L）, and a solution containing both TRAIL and mevastatin for 12, 24, 48 and 72 hours. MAIN OUTCOME MEASURES： Cell proliferation was detected using methyl thiazolyl tetrazolium assay; cell apoptosis was observed using Hoechst 33258 staining and fluorescence microscopy and was measured using Annexin V/propidium iodide flow cytometry; TRAIL R1/DR4 and TRAIL R2/DR5 protein expressions levels were measured using indirect immunofluorescence staining combined with flow cytometry in the recombinant soluble TRAIL （rsTRAIL, 300 tJg/L）, mevastatin （30 IJmol/L） and combination groups; TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression was detected using real-time polymerase chain reaction. RESULTS： rsTRAIL, mevastatin and their combination inhibited tumor proliferation in a time- and dose-dependent manner. The proliferation inhibitory rate and apoptosis rate of human SWO-38 cells in the combined group were significantly greater than the rsTRAIL or mevastatin alone group （P 〈 0.01）. TRAIL R1/DR4 and TRAIL R2/DR5 protein and mRNA expressions were increased in the combination group compared with mevastatin or rsTRAIL alone after 72 hours （P 〈 0.01）. CONCLUSION： Both rsTRAIL and mevastatin inhibit the proliferation and apoptosis of the human glioma cell line SWO-38, while their combination enhances the anti-tumor effect. The mechanism of action possibly correlates to the upregulation of TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression by mevastatin, thereby enhancing the cell sensitivity to rsTRAIL.

Expression of tumor necrosis factor related apoptosis inducing ligand receptor in glioblastoma

BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.

Molecular mechanism of action of anti-tumor necrosis factor antibodies in inflammatory bowel diseases

Anti-tumor necrosis factor(TNF) antibodies are successfully used in the therapy of inflammatory bowel diseases(IBD). However, the molecular mechanism of action of these agents is still a matter of debate. Apart from neutralization of TNF, influence on the intestinal barrier function, induction of apoptosis in mucosal immune cells, formation of regulatory macrophages as well as other immune modulating properties have been discussed as central features. Nevertheless, clinically effective anti-TNF antibodies were shown to differ in their mode-of-action in vivo and in vitro. Furthermore, the anti-TNF agent etanercept is effective in the treatment of rheumatoid arthritis but failed to induce clinical response in Crohn's disease patients, suggesting different contributions of TNF in the pathogenesis of these inflammatory diseases. In the following, we will review different aspects regarding the mechanism of action of anti-TNF agents in general and analyze comparatively different effects of each antiTNF agent such as TNF neutralization, modulation of the immune system, reverse signaling and induction of apoptosis. We discuss the relevance of the membranebound form of TNF compared to the soluble form for the immunopathogenesis of IBD. Furthermore, we review reports that could lead to personalized medicine approaches regarding treatment with antiTNF antibodies in chronic intestinal inflammation, by predicting response to therapy.

Autoimmune hepatitis and anti-tumor necrosis factor alpha therapy:A single center report of 8 cases

This article describes cases of anti-tumor necrosis factor(TNF)-α-induced autoimmune hepatitis and evaluates the outcome of these patients in relation to their immunosuppressive strategy. A retrospective analysis of medical records was performed in our center, in order to detect cases of autoimmune hepatitis(AIH) associated with anti-TNF biologic agents. We describe and analyze eight cases of AIH following anti-TNF therapy, 7 with infliximab and 1 with adalimumab. A distinction should be made between induction of autoimmunity and clinically evident autoimmune disease. Liver biopsy is useful in detecting the role of the TNF-α antagonist in the development of AIH. The lack of relapse after discontinuing immunosuppressive therapy favors, as in this case series, an immune-mediated drug reaction as most patients with AIH have a relapse after treatment is suspended. Although AIH related to anti-TNF therapy is rare, a baseline immunological panel along with liver function tests should be performed in all patients with autoimmune disease before starting biologics.

Total saponins of Aralia elata(Miq) Seem protect tumor necrosis factor-α-induced human umbilical vein endothelial cells via PI3K/Akt and NF-κB signal pathways

It is now thought that atherosclerosis,although due to enhanced lipid deposition,is mainly the result of a series inflammatory process.Total saponins of Aralia elata(Miq) Seem(TASAES) from the Chinese traditional herb Longya Araliachinensis L.,a folk medicine used for treating various diseases,increasing energy and improving the body′s ability to prevent hypoxia in Asian countries has attracted widespread attention.However,the ability of TASAES on inflammation-triggered vascular endothelial cell injury,a key early event in the pathogenesis of atherosclerosis,and its potential mechanisms of this protection have never been demonstrated.The present study determined the anti-inflammatory and anti-apoptoticactivities and protective mechanisms of the total aralosides of Araliaelata(Miq) Seem(TASAES) ameliorate tumor necrosis factor-α(TNF-α)-induced human umbilical vein endothelial cells(HUVECs) injury.Our results indicate that TASAES pretreatment provided cytoprotective effects by suppressing TNF-α-induced HUVECs apoptosis,mitochondrial membrane depolarization,caspase-3 activation,and modulation of inflammatory factors(IL-6,MCP-1 and VCAM-1),meanwhile inhibiting NF-κB transcription.Furthermore,the effect was correlated with the activation of the PI3K/Akt signal pathway.Blocking Akt activation with the PI3K inhibitor LY294002 effectively reversed the protective effect of TASAES against TNF-α-induced cell apoptosis.Moreover,the PI3K inhibitor partially blocked the effects of TASAES on the increasing of Bcl-2 and Bcl-xl protein expression,and inactivation of Bax protein expression.In conclusion,the results showed that TASAES decreased the inflammation and apoptosis of HUVECs caused by TNF-α treatment,and PI3K played a crucial role in enhancing cell sur.vival during this process.

Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in glioma U87 cells

Studies have shown that tumor necrosis factor-related apoptosis-inducing ligand（TRAIL）exhibits strong induction of apoptosis in human glioma cells.It remains unclear whether the mitochondrion pathway,an important apoptosis signaling pathway,is involved in TRAIL-induced glioma cell apoptosis.In the present study,in vitro cultured human glioma U87 cells were treated with human recombinant soluble TRAIL.Apoptosis of glioma U87 cells,mitochondrial transmembrane potential（Δψm）,cytoplasmic cytochrome c concentration and changes in caspase-3,-8 and-9 activity following human recombinant soluble TRAIL treatment were investigated to determine the mechanism of glioma U87 cell apoptosis induced by TRAIL.Additionally,blocking caspase-8resulted in TRAIL-induced mitochondrion pathway activation,suggesting that TRAIL,through activating caspase-8,initiated a series of mitochondrial events and resulted in apoptosis of glioma U87 cells.

血清sTWEAK、Netrin-1联合APACHEⅡ评分对重型颅脑损伤患者术后预后不良的预测价值

目的探讨血清可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)、神经轴突导向因子-1(Netrin-1)联合急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分对重型颅脑损伤患者术后预后不良的预测价值。方法选取2020年6月至2022年6月该院收治的120例重型颅脑损伤患者,根据术后30 d预后情况分为预后良好组和预后不良组。对比两组血清sTWEAK、Netrin-1水平及APACHEⅡ评分。采用单因素和多因素Logistic回归分析重型颅脑损伤患者术后预后不良的影响因素,并据以构建血清sTWEAK、Netrin-1及APACHEⅡ评分联合应用的预测模型,受试者工作特征(ROC)曲线分析血清sTWEAK、Netrin-1水平及APACHEⅡ评分对重型颅脑损伤患者术后预后不良的预测价值。结果预后不良组的重症监护室居住时间长于预后良好组,白蛋白水平、入院时格拉斯哥昏迷评分法评分和血清Netrin-1水平低于预后良好组,多发脑挫裂伤占比、机械通气占比、入院时APACHEⅡ评分和血清sTWEAK、血清肌酐、血尿素氮水平均高于预后良好组,差异有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,多发脑挫裂伤、Netrin-1水平降低、入院时APACHEⅡ评分升高、sTWEAK水平升高为重型颅脑损伤患者术后预后不良的危险因素(P<0.05)。ROC曲线分析结果显示,血清sTWEAK、Netrin-1及APACHEⅡ评分3个指标单独及联合应用时曲线下面积及其95%CI分别为0.742(0.552~0.925)、0.731(0.488~0.963)、0.714(0.502~0.911)、0.882(0.795~0.947)。结论血清sTWEAK、Netrin-1联合APACHEⅡ评分对重型颅脑损伤患者术后预后不良具有较好的预测价值,可为临床治疗方案的制订提供参考。

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。

Helicobacter pylori tumor necrosis factor-α inducing protein promotes cytokine expression via nuclear factor-κB

AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.

Tumor necrosis factor-α mediates JNK activation response to intestinal ischemia-reperfusion injury

AIM:To investigate whether tumor necrosis factor-α(TNF-α)mediates ischemia-reperfusion(I/R)-induced intestinal mucosal injury through c-Jun N-terminal kinase(JNK)activation.METHODS:In this study,intestinal I/R was induced by 60-min occlusion of the superior mesenteric artery in rats followed by 60-min reperfusion,and the rats were pretreated with a TNF-α inhibitor,pentoxifylline,or the TNF-α antibody infliximab.After surgery,part of the intestine was collected for histological analysis.The mucosal layer was harvested for RNA and protein extraction,which were used for further real-time polymerase chain reaction,enzyme-linked immunosorbent assay and Western blotting analyses.The TNF-α expression,intestinal mucosal injury,cell apoptosis,activation of apoptotic protein and JNK signaling pathway were analyzed.RESULTS:I/R significantly enhanced expression of mucosal TNF-α at both the mRNA and protein levels,induced severe mucosal injury and cell apoptosis,activated caspase-9/caspase-3,and activated the JNK signaling pathway.Pretreatment with pentoxifylline markedly downregulated TNF-α at both the mRNA and protein levels,whereas infliximab pretreatment did not affect the expression of TNF-α induced by I/R.However,pretreatment with pentoxifylline or infliximab dramatically suppressed I/R-induced mucosal injury and cell apoptosis and significantly inhibited the activation of caspase-9/3 and JNK signaling.CONCLUSION:The results indicate there was a TNFα-mediated JNK activation response to intestinal I/R injury.

血清Gal-3、TWEAK对精神分裂症的诊断价值及与精神症状严重程度的关系

目的研究血清半乳糖凝集素3(Gal-3)、肿瘤坏死因子样弱凋亡诱导因子(TWEAK)对精神分裂症的诊断价值及与精神症状严重程度的关系。方法选取2019年3月至2021年3月在该院诊治的96例精神分裂症患者作为病例组,以同期60例健康体检者为对照组,进行回顾性分析。采用酶联免疫吸附试验检测血清Gal-3、TWEAK水平。采用Pearson相关分析血清Gal-3、TWEAK水平与临床指标的相关性;采用多因素Logistic回归分析影响精神分裂症发生的因素;采用受试者工作特征(ROC)曲线分析血清Gal-3、TWEAK单项及联合检测对精神分裂症的诊断价值。结果观察组PANSS评分总分、阳性症状评分、阴性症状评分、一般精神病理学症状评分及血清Gal-3、TWEAK水平均高于对照组,差异均有统计学意义(P<0.001)。血清Gal-3、TWEAK水平与PANSS评分总分、阳性症状评分、阴性症状评分、一般精神病理学症状评分呈正相关(P<0.001)。PANSS评分总分、阳性症状评分、一般精神病理学症状评分升高及血清Gal-3、TWEAK水平升高均是精神分裂症发生的独立危险因素(P<0.001)。血清Gal-3、TWEAK联合检测诊断精神分裂症的曲线下面积为0.870(95%CI:0.831~0.919),明显高于血清Gal-3、TWEAK单项检测诊断精神分裂症的AUC[0.812(95%CI:0.769~0.847)、0.820(95%CI:0.771~0.852)],差异均有统计学意义(Z=4.345,P<0.001;Z=4.010,P=0.002)。结论精神分裂症患者血清Gal-3、TWEAK水平升高,二者与精神症状严重程度有关,血清Gal-3、TWEAK联合检测对精神分裂症具有较高的诊断价值。

T2DM患者血清sTWEAK、sCD40L水平与糖尿病微血管并发症的相关性分析

目的 探讨2型糖尿病(T2DM)患者血清可溶性肿瘤坏死因子(TNF)样凋亡弱诱导因子(sTWEAK)、可溶性CD40配体(sCD40L)水平与糖尿病微血管并发症(DMA)的相关性。方法 选取2021年3月-2023年3月航天中心医院收治的150例T2DM患者为研究对象,根据患者是否合并DMA分为DMA组(n=69)及T2DM组(n=81)。检测并比较两组患者血清sTWEAK、sCD40L水平及临床资料,采用Logistic回归分析影响T2DM并发微血管病变的危险因素,绘制受试者工作特征曲线(ROC)分析sTWEAK、sCD40L对T2DM并发微血管病变的预测价值。结果 DMA组患者血清sTWEAK、sCD40L水平均高于T2DM组(P<0.05);DMA组的病程、FPG及HbA1c、sTWEAK、sCD40L水平均高于T2DM,差异均有统计学意义(P<0.05);Logistic分析结果显示病程长、HbA1c高水平、sTWEAK高水平及sCD40L高水平均是导致T2DM并发微血管病变的独立危险因素(P<0.05)。ROC结果显示,血清sTWEAK、sCD40L单独及联合预测T2DM并发微血管病变的AUC(95%CI)分别为0.714(0.635~0.785)、0.744(0.666~0.812)、0.859(0.792~0.910),二者联合的预测效能优于单独检测(Z=2.626、2.395,P<0.05)。结论 血清sTWEAK、 sCD40L在DMA患者中均异常升高,与DMA关系密切,sTWEAK、sCD40L联合对于T2DM并发微血管病变的预测价值较高。

Role of tumor necrosis factor-like weak inducer of apoptosis （TWEAK）/fibroblast growth factor-inducible 14 （Fnl4） axis in rheumatic diseases

Tumor necrosis factor （TNF）-Iike weak inducer of apoptosis （TWEAK） is a member of the TNF superfamily of structurally related cytokines and is known to induce proliferation, migration, differentiation, apoptotic celt death, inflammation, and angiogenesis. These physiological processes are induced by the binding of TWEAK to fibroblast growth factor-inducible 14 （Fn14）, a highly inducible cell-surface receptor that is linked to several intracellular signaling pathways, including the nuclear factor-KB （NF-KB） pathway. This review discusses the role of the TWEAK-Fn14 axis in several rheumatic diseases and the potential therapeutic benefits of modulation of the TWEAK-Fn14 pathway.

Targeted anti-cancer therapy: Co-delivery of VEGF siRNA and Phenethyl isothiocyanate (PEITC) via cRGD-modified lipid nanoparticles for enhanced anti-angiogenic efficacy

Anti-tumor angiogenesis therapy, targeting the suppression of blood vessel growth in tumors, presents a potent approach in the battle against cancer. Traditional therapies have primarily concentrated on single-target techniques, with a specific emphasis on targeting the vascular endothelial growth factor, but have not reached ideal therapeutic efficacy. In response to this issue, our study introduced a novel nanoparticle system known as CS-siRNA/PEITC&L-cRGD NPs. These chitosan-based nanoparticles have been recognized for their excellent biocompatibility and ability to deliver genes. To enhance their targeted delivery capability, they were combined with a cyclic RGD peptide (cRGD). Targeted co-delivery of gene and chemotherapeutic agents was achieved through the use of a negatively charged lipid shell and cRGD, which possesses high affinity for integrin αvβ3 overexpressed in tumor cells and neovasculature. In this multifaceted approach, co-delivery of VEGF siRNA and phenethyl isothiocyanate (PEITC) was employed to target both tumor vascular endothelial cells and tumor cells simultaneously. The co-delivery of VEGF siRNA and PEITC could achieve precise silencing of VEGF, inhibit the accumulation of HIF-1α under hypoxic conditions, and induce apoptosis in tumor cells. In summary, we have successfully developed a nanoparticle delivery platform that utilizes a dual mechanism of action of anti-tumor angiogenesis and pro-tumor apoptosis, which provides a robust and potent strategy for the delivery of anti-cancer therapeutics.

前列腺癌患者血清TWEAK和SREBP-1水平表达与临床病理特征及无进展生存预后的关系研究

目的 研究前列腺癌(prostate cancer,PC)患者血清肿瘤坏死因子样弱凋亡诱导因子(tumor necrosis factor like weak inducer of apoptosis,TWEAK)、固醇调节元件结合蛋白1(sterol regulatory element-binding protein 1,SREBP-1)表达与临床病理特征及无进展生存预后的关系。方法 选取2018年1月~2020年1月武汉市东西湖区人民医院行PC根治术的94例PC患者为PC组,以同期50例前列腺增生(benign prostatic hyperplasia,BPH)患者为BPH组,以同期体检的50例健康人为对照组。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测血清TWEAK和SREBP-1表达水平。Kaplan-Meier生存分析比较血清TWEAK和SREBP-1对PC患者无进展生存预后的影响。多因素COX回归分析影响PC患者无进展生存预后的因素。结果PC组患者血清TWEAK(77.14±15.46 ng/L),SREBP-1(334.14±33.81 ng/L)高于BPH组(38.69±10.58 ng/L,201.69±28.74 ng/L)和对照组(36.26±10.27 ng/L,189.51±27.65 ng/L),差异具有统计学意义(t=23.752,25.249;34.636,37.821,均P<0.05)。PC患者血清TWEAK与SREBP-1表达呈显著正相关(r=0.668,P=0.001)。Gleason评分> 7分、TNM分期Ⅲ期及术前前列腺特异抗原(prostate specific antigen,PSA)水平≥20 ng/ml的PC患者血清TWEAK,SREBP-1水平高于Gleason评分≤7分,TNM分期Ⅰ~Ⅱ期及术前PSA水平<20 ng/ml,差异具有统计学意义(t=8.465~16.597,均P<0.05)。TWEAK高表达组和低表达组三年总体无进展生存率分别为60.42%(29/48)和86.96%(40/46),SREBP-1高表达组和低表达组三年总体无进展生存率分别为57.78%(26/45)和87.76%(43/49);TWEAK高表达组、SREBP-1高表达组三年累积无进展生存率低于TWEAK低表达组、SREBP-1低表达组,差异具有统计学意义(Log-rankχ2=8.125,9.547,P=0.004,0.002)。TNM分期Ⅲ期(OR=1.448,P <0.001)、Gleason评分> 7分(OR=1.401,P <0.001)、术前PSA≥20 ng/ml (OR=1.353,P <0.001)及血清TWEAK (OR=1.338,P <0.001)和SREBP-1 (OR=1.293,P <0.001)是影响PC患者无进展生存预后的独立危险因素。结论 PC患者血清TWEAK和SREBP-1升高,两者与PC临床病理特征相关,是评估无进展生存预后的血清标志物。

2型糖尿病患者血清PTX3、sTWEAK水平与非酒精性脂肪性肝病的关系研究

目的探讨2型糖尿病(T2DM)患者血清正五聚蛋白3(PTX3)、可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)水平与非酒精性脂肪性肝病(NAFLD)的关系。方法选取北部战区总医院152例新诊断为T2DM的患者为研究对象,根据是否合并NAFLD将患者分为NAFLD组(92例)和非NAFLD组(60例);根据肝脏超声检查结果,将NAFLD患者分为轻度组、中度组和重度组。另选取35例健康人作为对照(对照组)。采用酶联免疫吸附试验检测血清PTX3、sTWEAK水平。采用Pearson相关分析T2DM患者PTX3和sTWEAK水平与总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、空腹血糖(FPG)、糖化血红蛋白(HbA1c)、胰岛素抵抗指数(HOMA-IR)的关系。采用多因素Logistic回归分析T2DM合并NAFLD的影响因素;采用受试者工作特征(ROC)曲线分析PTX3、sTWEAK对NAFLD的预测价值,计算曲线下面积(AUC)。比较轻度组、中度组、重度组血清PTX3、sTWEAK水平。结果与对照组比较,NAFLD组和非NAFLD组血清PTX3、sTWEAK水平较高(P<0.05)。与非NAFLD组比较,NAFLD组血清PTX3、sTWEAK水平较高(P<0.05)。体质量指数(BMI,OR=3.387)、TG(OR=1.958)、HOMA-IR(OR=3.040)、PTX3(OR=4.836)、sTWEAK(OR=4.133)是T2DM合并NAFLD的影响因素(P<0.05)。T2DM患者血清PTX3水平分别与BMI、LDL-C、FPG、HbA1c、HOMA-IR呈正相关(P<0.05),而与HDL-C呈负相关(P<0.05)。血清sTWEAK水平分别与LDL-C、FPG、HOMA-IR呈正相关(P<0.05)。血清PTX3、sTWEAK预测T2DM患者发生NAFLD的AUC分别为0.873和0.821,二者联合可将AUC提高至0.915。轻度组、中度组、重度组血清PTX3和sTWEAK水平比较,差异有统计学意义(P<0.05),病情越重,患者血清PTX3和sTWEAK水平越高。结论PTX3、sTWEAK是T2DM患者发生NAFLD的影响因素,并且血清PTX3、sTWEAK水平越高,NAFLD病情越重。

Effect of hepatocyte apoptosis induced by TNF-α on acute severe hepatitis in mouse models

AIM To study the effect of hepatocyteapoptosis and necrosis induced by TNF-α on thepathogenesis of acute severe hepatitis(ASH).METHODS The model of ASH was prepared inD-galactosamine(GAIN)sensitized BALB/c miceby injection of either endotoxin(ET)or tumornecrosis factor-α(TNF-α).Morphologicalchanges of apoptotic hepatocytes were studiedby both light and electron microscope and in siteend labeling method(ISEL).Molecular biologicalchanges of DNA ladder were observed byelectrophoresis of extract from liver tissues.Biochemical changes were measured by alanineaminotransferase(ALT),asparticaminotransferase(AST)and TNF-α.The relationbetween apoptosis and necrosis was evaluatedsimultaneously.RESULTS The sequence of hepatocyteapoptosis,necrosis,and final death from ASHwas observed both in GAIN/ET and GAIN/TNF-agroup.Apoptosis was prominent at 3.5 h and 5 hafter injection of inducer,while necrosis becamedominant at 9 h after challenge.The appearanceof apoptosis was earlier in GAIN/TNF-α groupthan that in GAIN/ ET group.Pretreatment ofmice with antiTNF IgG1 may completely preventthe liver injury induced by GalN/ET.CONCLUSION TNF-α can cause liver damageby inducing hepatic apoptosis and necrosis inmice with endotoxemia.

Increase of TNFα-stimulated Osteoarthritic Chondrocytes Apoptosis and Decrease of Matrix Metalloproteinases 9 by NF-κB Inhibition

Objective To investigate the in vitro effect of caffeic acid phenethyl ester （CAPE）, a NF-KB inhibitor, on the apoptosis of osteoarthritic （OA） chondrocytes and on the regulation of the gelatinases matrix metalloproteinase 2 （MMP-2） and matrix metalloproteinase 9 （MMP-9）. Methods Annexin V-FITC/propidium iodide （PI） labeling and western blotting were used to observe and determine the apoptosis in TNFa-stimulated primary cultured osteoarthritic chondrocytes. Also, gelatin zymography was applied to examine MMP-2 and MMP-9 activities in supernatants. Results it was confirmed by both flow cytometry and western blotting that chondrocytes from OA patients have an apoptotic background. Use of CAPE in combination with 10 ng/mL of TNFa for 24 h facilitated the apoptosis. MMP-9 in the supernatant could be autoactivated （from proMMP-9 to active MMP-9）, and the physiologic calcium concentration （2.5 mmol/L） could delay the autoactivation of MMP-9. The activities of MMP-2 and MMP-9 in the fresh supernatant increased significantly in response to stimulation by 10 ng/mL of TNFa for 24 h. The stimulatory effect of TNFa just on proMMP-9 was counteracted significantly by CAPE. Conclusion NF-KB could prevent chondrocytes apoptosis though its activation was attributed to the increase of proMMP-9 activity induced by TNFa （a pro-apoptotic factor）. Therefore, therapeutic NF-KB inhibitor was a ＇double-edged swords＇ to the apoptosis of chondrocytes and the secretion of MMP-9.