重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白靶向治疗类风湿关节炎的临床疗效观察

目的 观察重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白（益赛普）联合甲氨蝶呤（MTX）治疗类风湿关节炎的临床疗效观察及安全性.方法 类风湿关节炎患者60例,随机分为两组.实验组30例,给予每周两次益赛普25mg注射,联合甲氨蝶呤（MTX）治疗12周;对照组30例,单用MTX治疗12周.结果 实验组有效率96.7%,对照组有效率66.7%,实验组与对照组比较差异有统计学意义（P＜0.05）.实验组在改善临床症状指标方面明显优于对照组（P＜0.05）,两组不良反应发生率比较无显著统计学意义.结论 益赛普联合MTX治疗与单用MTX相比可明显改善类风湿关节炎临床症状和实验室指标,起效迅速,疗效显著,且不良反应无明显增加.     

重组人Ⅱ型肿瘤坏死因子受体抗体融合蛋白对强直性脊柱炎患者疗效及炎症因子、脊柱活动度的影响

目的探讨重组人Ⅱ型肿瘤坏死因子受体抗体融合蛋白(rhTNFR:Fc)对强直性脊柱炎(AS)患者疗效及炎症因子、脊柱活动度的影响。方法纳入2015-02-2017-02,于我院治疗的76例AS患者,随机均分为观察组与对照组各38例。对照组患者给予甲氨蝶呤、柳氮磺吡啶治疗,观察组在此基础上联合rhTNFR:Fc皮下注射,连续治疗3个月。比较两组治疗前后血清肿瘤坏死因子α(TNF-α)、C反应蛋白(CRP)等炎性因子水平、chober试验、颈部旋转等脊柱活动度指标以及脊柱疼痛、关节肿胀等症状指标,评价治疗3各月后综合疗效并记录不良反应发生情况。结果两组治疗后,血清TNF-α、CRP、白细胞介素6(IL-6)、IL-8水平均显著降低,且治疗后观察组显著低于对照组,差异有统计学意义(P0.05);两组治疗后chober试验、颈部旋转、腰椎侧弯、扩胸度均显著增加,且治疗后观察组显著高于对照组,差异有统计学意义(P0.05);两组治疗后AS病情活动指数(BASDAI)、晨僵时间、脊柱疼痛评分均显著降低,且治疗后观察组显著低于对照组,差异有统计学意义(P0.05);观察组治疗总有效率为89.47%,显著高于对照组的76.32%,差异有统计学意义(P0.05)。两组不良反应发生率差异无统计学意义(P0.05)。结论 rhTNFR:Fc皮下注射联合甲氨蝶呤、柳氮磺吡啶治疗AS,能显著改善患者临床症状及脊柱活动度,降低炎症反应,治疗安全有效。     

肿瘤坏死因子受体-抗体融合蛋白治疗类风湿关节炎患者的护理

对30例类风湿关节炎(RA)患者采用肿瘤坏死因子受体-抗体融合蛋白(TNFR:FC)进行治疗,并加强心理护理、用药护理及不良反应的护理.结果 显效24例,有效4例,总有效率93.3%.提示护理干预可减轻RA患者药物不良反应,提高治疗舒适度,从而提高疗效.     

重组人II型肿瘤坏死因子受体-抗体融合 蛋白对胶原诱导性关节炎大鼠骨质及 骨保护素的影响

目的研究重组人H型肿瘤坏死因子受体-抗体融合蛋白（rhTNFR:Fc，益赛普）及其他慢作用 药对胶原诱导性关节炎大鼠骨密度及骨保护素的影响，探讨各种方案对骨代谢生化指标、骨质的改 变。方法建立H型胶原诱导的雄性Wislar大鼠CIA模型，随机分为正常对照组、CIA模型对照组、MTX联合强的松龙组、MTX联合羟氯喹和柳氮磺吡啶组、MTX联合益赛普组。分别于第10、21周予 行大鼠四肢关节钼靶拍片及测骨保护素（韵孕郧)。结果 CIA组大鼠第10周出现骨质疏松、关节软 骨、骨破坏、关节间隙狭窄，MTX + pred组大鼠出现骨质疏松，关节腔结构尚完整，MTX + HCQ + 杂杂在 组大鼠出现骨质疏松、关节软骨、骨轻度破坏、关节间隙轻度狭窄，MTX + rhTNFR：Fc组大鼠则仅出 现骨质轻度破坏；第21周时CIA组、MTX + pred组和MTX + HCQ + SSZ组大鼠骨质破坏加重，而MTX + rhTNFR:Fc组大鼠则未见骨质破坏加重。源组血清韵孕郧水平在治疗前均出现降低，第10周时4 组韵孕郧水平均出现降低，但MTX + rhTNFR:Fc组降低较其他3组有统计学意义（孕<0.05 )。21周 时CIA组、MTX + pred组和MTX + HCQ + SSZ组韵孕郧水平均出现进一步降低（孕<0.05 )，但MTX + rhTNFR:Fc组未见明显降低（P >0.05 )。结论rhTNFR:Fc能有效抑制胶原诱导性关节炎大鼠骨质 破坏，其作用机制可能与韵孕郧相关。     

运动疗法联合重组人Ⅱ型肿瘤坏死因子受体抗体融合蛋白治疗强直性脊柱炎的疗效观察

目的探索运动疗法与重组人Ⅱ型肿瘤坏死因子受体抗体融合蛋白(rh TNFR:Fc)联合应用于强直性脊柱炎的治疗效果。方法抽取2016-04—2017-07间焦作市第二人民医院骨科收治的118例强直性脊柱炎患者,根据患者意愿分组。对照组接受rh TNFR:Fc皮下注射。观察组在对照组基础上给予运动疗法。比较2组的DAI指数(身体疲倦程度、关节疼痛程度以及关节僵硬程度)、C反应蛋白(CRP)水平、肿瘤坏死因子-α(TNF-α)和红细胞沉降率(ESR)等指标。结果治疗前2组的各项指标差异无统计学意义(P0.05)。治疗12周后,观察组患者的身体疲倦程度、关节疼痛程度及关节僵硬程度评分低于对照组,且关节晨僵时间短于对照组,差异有统计学意义(P0.05)。在CRP、ESR、TNF-α水平检测中,观察组均低于对照组,差异有统计学意义(P0.05)。结论强直性脊柱炎患者在应用rh TNFR:Fc治疗的基础上给予运动疗法,有助于提高治疗效果,缓解临床症状,提高生活质量。     

注射用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白联合沙利度胺对强直性脊柱炎患者血清炎性因子的影响

目的探讨注射用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(recombinant human type II tumor necrosis factor receptor-antibody fusion protein,rh TNFR:Fc)联合沙利度胺对强直性脊柱炎(ankylosing spondylitis,AS)患者血清炎性因子的影响。方法选取2016年3月~2018年10月经本院诊治的AS患者82例,随机分为研究组和对照组各41例。对照组患者给予沙利度胺口服治疗,研究组患者在对照组的基础上加用rh TNFR:Fc皮下注射治疗,两组均持续治疗12周。比较两组患者治疗前及治疗后6周、12周的血清红细胞沉降率(erythrocyte sedimentation rate,ESR)、C反应蛋白(C-reactive protein,CRP)及肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)水平及临床症状。记录两组患者临床疗效及不良反应发生情况。结果研究组患者临床总有效率为95. 12%,明显高于对照组的80. 49%(P!0. 05);治疗后6周、12周,两组患者ESR、CRP、TNF-α水平均明显低于治疗前,但研究组三项指标水平均明显低于对照组(P!0. 05);治疗后6周、12周,两组患者临床各指标均较治疗前明显改善,但研究组患者临床各指标改善效果明显优于对照组(P!0. 05);两组患者并发症发生率的比较,无统计学差异(P"0. 05)。结论rh TNFR:Fc联合沙利度胺可明显改善机体炎症反应,有效缓解临床症状,具有显著疗效。     

Use of porcine tumor necrosis factor receptor 1-Ig fusion protein to prolong xenograft survival

BACKGROUND: Delayed rejection of xenografts is a major hurdle that needs to be addressed to achieve long-term engraftment in the pig-to-primate transplant setting. Both vascular and avascular xenografts are susceptible to a delayed rejection process that comprises humoral and cellular responses. Tumor necrosis factor (TNF) is believed to play a role in this process by promoting cell activation, apoptosis and the recruitment of inflammatory cells. To address this problem, we engineered the donor cell in such a way that it could block both human and porcine TNF. METHODS: We produced a recombinant fusion protein containing the extracellular domain of the porcine TNF-Receptor 1 and an IgG Fc moiety (pTNFR1Ig). We first evaluated by flow cytometry the pTNFR1Ig capacity to prevent TNF alpha-induced expression of SLAI, SLAII, VCAM-1, ICAM-1 and E-selectin on the cell surface of porcine aortic endothelial cells (PAEC). The effect on TNF alpha-mediated cell death was also assessed by propidium iodide staining after incubating PAEC with TNF alpha plus cycloheximide for 24 h. PAEC and porcine fibroblasts were subsequently engineered by retroviral infection to express and secrete pTNFR1Ig and their resistance to the TNF alpha effects was tested in vitro. Finally, we transplanted mock-control and pTNFR1Ig-expressing PAEC under the kidney capsule of BALB/c mice in the absence of immunosuppression and examined the degree of rejection at 2 and 3 weeks post-transplantation. RESULTS: Treatment with pTNFR1Ig resulted in a very potent blockade of human, porcine and murine TNF alpha activity on porcine cells. It inhibited the upregulation of all cell surface markers of activation tested as well as the TNF alpha-mediated cell death. Moreover, pTNFR1Ig-expressing PAEC showed prolonged engraftment in a pig-to-mouse xenotransplant model. CONCLUSIONS: Incorporation of strategies that block TNF may prove useful in the development of xenografts resistant to delayed rejection.     

Antitumor effects of recombinant human tumor necrosis factor against human tumor xenografts transplanted into nude mice

Antitumor activities of recombinant human tumor necrosis factor (rH-TNF) against human tumor xenografts in nude mice were studied. Thirteen human tumor xenografts serially transplanted into nude mice were used for experiments; five gastric, two breast, two gallbladder, one colon and one esophageal carcinoma, one liposarcoma and one squamous carcinoma of the neck. They were inoculated into the subcutaneous tissue of BALB/c nu/nu nude mice and the treatment was started when the estimated tumor weight reached 100–300 mg. rH-TNF was administered intratumorally at schedule of qd×5 or q3d×5. rH-TNF showed a marked antitumor activity against various human tumors. The hemorrhagic necrosis was observed in all types of the human tumor xenografts (100 per cent), and the complete regression of the tumor was noted in 4 of 11 tumors (36.4 per cent). On the contray, intraperitoneal rH-TNF exhibited little antitumor effect. The additive effect in the combination of TNF and Mitomycin C was observed against two Mitomycin C resistant gastric tumors.     

重组人肿瘤坏死因子抗结肠癌的体外研究

研究重组人肿瘤坏死因子（ｒｈＴＮＦ）体外对人结肠癌细胞株ＳＷ－４８０的抗瘤作用，改良ＭＴＴ地测试表明，ｒｈＴＮＦ在较高剂量时才能瘤细胞表现细胞毒性，而联合应用５－氟脲嘧或丝裂霉素，ｒｈＴＮＦ在很小剂量即可获得增强细胞毒性，＾３Ｈ－ＴｄＲ掺入率测定表明ｒｈＴＮＦ细胞毒性与瘤细胞ＤＮＡ合成受抑具有一致性关系，电镜观察发现，ｒｈＴＮＦ作用瘤细胞先发生线粒体及细胞核的破坏，然后出现细胞膜及细胞裂解。     

Enhanced in vivo cytotoxicity of recombinant human tumor necrosis factor with etoposide in human renal cell carcinoma

Summary The combination of tumor necrosis factor (TNF) and etoposide (ETP) was evaluated for potential cytotoxic efficacy against a human renal cell carcinoma xenograft using an in vivo assay employing an athymic mouse host with tumor implanted a the subrenal capsule site. Both antitumor efficacy (relative survival or RTS) and toxicity (weight loss) of TNF and ETP alone and in combination were evaluated. While TNF and ETP alone were mildly inhibitory (RTS 90% and 71%, respectively), the combination caused marked tumor inhibition (45% of controls). Host toxicity encountered with the combination did not exceed the toxicity associated with ETP alone, suggesting that the therapeutic index may have been augmented. It is concluded that enhanced antitumor activity without substantial augmentation of toxicity is observed with this combination, providing a rationale for further evaluation of tumor necrosis factor-based regimens for the treatment of advanced renal carcinoma.Supported by a Merit Review grant, VA Medical Research Service, Durham, NC 27710, USA     

Differential sensitivity of hormone-responsive and unresponsive human prostate cancer cells (LNCaP) to tumor necrosis factor

Summary Two sublines, the hormone-sensitive LNCaP-FGC and the insensitive LNCaP-r (resistant) carcinoma cell lines, originating from the parental human prostatic carcinoma cell line LNCaP were tested for sensitivity to human tumor necrosis factor- (TNF) using the MTT assay. Irrespective of the culture conditions, i.e., whether FGC cell growth was hormone stimulated or hormone deprived, a clear dose-related response was observed between the concentration of TNF (range: 5–5000 U/ml) in the culture medium and the percentage of growth inhibition. In medium containing androgen-depleted serum, in which FGC cells showed reduced proliferative activity, the percentage of inhibition by a concentration of 100 U/ml TNF was substantially higher than that found in hormone-stimulated cells (90% and 60%, respectively). In contrast to the FGC cells, the hormone-insensitive LNCaP-r cells were almost completely resistant to the action of TNF. Growth of the FGC cells was almost completcly inhibited, whereas growth of the LNCaP-r cells was retarded with only 20% at dosages up to 5000 U/ml. This substantial difference in TNF responsiveness could not be ascribed to differences in TNF-binding capacity, as both the FGC and LNCaP-r cells were found to contain identical numbers of TNF-receptors (approximately 1000 sites/cell). A possible association between hormone responsiveness and TNF sensitivity is suggested for these LNCaP sublines.This study was supported in part by the Stichting Urologisch Wetenschappelijk Onderzoek (SUWO)This paper was selected for publication in Urological Research from the program of the 1991 meeting of the European Society of Urological Oncology and Endocrinology (ESUOE)     

肿瘤坏死因子基因转染对人胆管癌细胞生长的影响

目的　研究肿瘤坏死因子 α(TNF α)基因过表达对人胆管癌细胞生长的影响。方法构建逆转录病毒 TNF α(pLNCX TNF α)复合体 ,转染人胆管癌细胞得到稳定表达 ,通过聚合酶链式反应 (PCR)、流式细胞仪 (FCM )、免疫组织化学等方法检测基因表达、细胞生长增殖和细胞凋亡等情况。结果　pLNCX TNF α基因转染人胆管癌细胞后 ,pLNCX空质粒转染组、pLNCX TNF α基因转染组的细胞均能扩增出 70 8bp的基因片断 ,而未转染组则无 ;pLNCX TNF α基因转染组的细胞克隆抑制率为 64 % ,与对照组和 pLNCX空质粒转染组相比 ,差异性非常显著 (P <0 .0 1) ;转染的人胆管癌细胞G1期比例从 2 9.0 7%增加到 5 4.2 4% ,G2 M期和S期比例分别从 5 2 .0 3 %下降到2 .0 2 %、18.3 2 %下降到 10 .3 4% ;凋亡细胞数从 5 .83 %增加到 3 4.76%。结论　TNF α基因通过诱导肿瘤细胞发生凋亡并导致其发生G1期阻滞在肿瘤基因治疗方面发挥作用。     

TNF-α拮抗剂(Etanercept)治疗脊髓损伤作用机制的实验研究

目的探讨TNF-α拮抗剂Etanercept(依那西普)对继发性脊髓损伤的治疗作用和可能机制。方法建立大鼠脊髓挫伤模型,损伤后1 h对大鼠行腹腔注射5 mg/kg Etanercept或生理盐水(损伤对照组);假手术组(20只大鼠)仅行椎板切除术而无脊髓损伤,1 h后给予腹腔注射1 ml生理盐水。结果在脊髓损伤急性期(12 h、1 d、3 d),Etanercept治疗组TNF-α的蛋白表达强度明显弱于损伤对照组。与对照组相比,Etanercept治疗组TNFR1的表达在损伤后6、12 h均下降,TNFR2仅在损伤后6 h下降。损伤对照组大鼠在脊髓损伤后后肢运动功能明显受限,而Etanercept治疗组大鼠在脊髓损伤后2、4、8周,后肢BBB评分显著升高。结论 Etanercept可能通过抑制TNF-α/TNFR通路,减轻了脱髓鞘变性,促进了运动功能的恢复。     

人三叶因子1融合蛋白对烧伤小鼠胃黏膜的保护作用

目的 了解人三叶因子1(hTFF1)融合蛋白在减轻烧伤小鼠胃黏膜损害中的作用.方法 利用原核重组表达质粒pET32α-hTFF1,诱导表达基因重组hTFF1(rhTFF1)融合蛋白.采用小鼠30%TBSAⅢ度烧伤模型,按照随机数字表法分为烧伤治疗组:给予一定时间和一定剂量的rhTFF1融合蛋白灌胃治疗;烧伤对照组:方式同前,改用等渗盐水灌胃.另设正常对照组.通过胃黏膜外观、损伤指数和病理切片结果,比较前2组小鼠胃黏膜损伤前后的情况,并以正常对照组相应指标为参照. 结果 能够获得具有良好特异性的rhTFF1融合蛋白.伤后1 d小鼠胃黏膜损伤达峰值,烧伤治疗组和烧伤对照组小鼠胃黏膜糜烂发生率分别为22.2%和77.8%,损伤指数分别为2.0±1.2和6.2±2.0.正常对照组小鼠上述指标为0. 结论 rhTFF1融合蛋白能够在大肠杆菌系统表达,对小鼠烧伤后损害的胃黏膜有明显的治疗作用.     

The antitumor effects of locally injecting human peripheral blood mononuclear cells treated with OK-432 into the tumor site: The possible role of a tumor growth inhibitory factor (TGIF)

The production of a tumor growth inhibitory factor (TGIF) was induced in human peripheral blood mononuclear cells (PBMC) by a streptococcal preparation, OK-432,in vitro. The antitumor effect of locally injecting PBMC treated with OK-432 into the tumor site was studied. PBMC were collected from patients with gastric cancer 5 to 12 days before their operation, and cultured with OK-432 for 24 hrin vitro. After the culture, the PBMC were washed thoroughly to eliminate the OK-432. The washed PBMC went on producing TGIF for more than 72 hrin vitro in the absence of OK-432. A small number of TGIF-producing PBMC, approximately 10⁷ cells, were injected around the lesion under endoscopic observation. A remarkable antitumor effect was observed in 2 out of 10 cases of resectable gastric cancer. Histological examinations indicated that the antitumor effect is due to antitumor cytokines such as TGIF produced by PBMC rather than to the OK-432-activated PBMC themselves.     

Sensitivity of human complement factor H related protein (BTA stat) test and voided urine cytology in the diagnosis of bladder cancer

PURPOSE: We compared the sensitivity of the BTA statdagger test, a rapid, noninvasive, qualitative urine test that detects bladder tumor associated antigen (human complement factor H related protein) in urine, to that of voided urine cytology in patients with primary bladder cancer. We also assessed the effect of tumor size, number, histological grade and stage on test sensitivity. MATERIALS AND METHODS: We evaluated 151 patients with newly diagnosed bladder cancer in a prospective multicenter study. A voided urine sample obtained before transurethral bladder tumor resection was divided for culture, cytology and BTA stat testing. RESULTS: Overall sensitivity of the BTA stat test and urine cytology for detecting primary bladder cancer was 81.5% and 30.3%, respectively (p <0.0001). The sensitivity of each test increased as tumor size, number, histological grade and stage increased. CONCLUSIONS: Sensitivity of the BTA stat test was superior to that of voided urine cytology in all tumor categories. This noninvasive, easy to perform, point of care test may have the potential to replace cytology for diagnosing bladder cancer.