高糖高脂对系膜细胞产生细胞外基质及PAF的影响

目的： 探讨高糖高脂对系膜细胞产生细胞外基质（ECM）和血小板活化因子（PAF）的影响。方法：人系膜细胞单独培养,分为对照组、甘露醇组、高糖高脂组、PAF受体拮抗剂BN52021+高糖高脂组, ELISA法检测各组细胞上清液中纤维连接蛋白（Fn）、IV型胶原（Col IV）、血小板活化因子（PAF）含量,实时荧光定量检测系膜细胞血小板活化因子受体（PAF-R） mRNA表达。结果：高糖高脂可引起系膜细胞培养上清液Fn和Col IV含量升高（均P<0.05）,BN52021可抑制高糖高脂引起的Fn和Col IV含量升高（P<0.05）;高糖高脂可引起系膜细胞培养上清液PAF含量升高（P<0.05）;高糖高脂可上调系膜细胞PAF-R mRNA表达（P<0.05）。结论: 高糖高脂刺激系膜细胞Fn、Col IV、PAF产生,高糖高脂刺激的ECM分泌增加部分与PAF有关; 高糖高脂可促进系膜细胞PAF-R基因表达,使PAF的生物效应进一步放大。     

川芎嗪对实验性肾炎肾皮质内血小板活化因子、血栓素B_2的影响

目的 :探讨川芎嗪对实验性肾炎肾皮质内血小板活化因子 (PAF)、血栓素B2 (TXB2 )水平的影响。方法 :复制大鼠加速型抗肾小球基膜 (GBM)肾炎模型。分为肾炎组、肾炎 川芎嗪组 (治疗组 )。结果 :治疗组各期尿蛋白量较肾炎组减少 (P <0 .0 5 ,P <0 .0 1) ,第 2 1d时血清肌酐含量低于肾炎组 (P <0 .0 5 ) ,光镜和电镜下肾脏病理改变较肾炎组为轻 ,同时肾皮质内PAF、TXB2 含量少于肾炎组 (P <0 .0 1)。结论 :川芎嗪治疗实验性肾炎有效 ,其机理可能与减少肾皮质内PAF、TXB2 等有关。     

血小板活化因子受体拮抗剂对实验性肾病综合征的疗 …

目的：探讨血小板活化因子受体拮抗剂对实验性肾病综合征的疗效机制。方法：实验大鼠均复制成阿霉素（ＡＤＲ）肾病模型,分为２组：ＡＤＲ肾病组和ＡＤＲ肾病＋血小板活化因子（ＰＡＦ）受体拮抗剂（ＢＮ５２０２１）组。结果：ＡＤＲ肾病＋ＢＮ４２０２１组大鼠各期尿蛋白量、血清总蛋白下了幅度、血清胆固醇上升幅度均显著低于ＡＤＲ肾病且（Ｐ〈０．０５）,第２１天时血清肌酐含量显著低于ＡＤＲ肾病组（Ｐ〈０．０５）,电镜下     

川芎嗪对实验性肾炎肾皮质内血小板活化因子、血栓素B2的影响

目的：探讨川芎嗪对实验性肾炎肾皮质内血小板活化因子（PAF）、血栓素B₂（TXB₂）水平的影响。方法：复制大鼠加速型抗肾小球基膜（GBM）肾炎模型。分为肾炎组、肾炎+川芎嗪组（治疗组）。结果：治疗组各期尿蛋白量较肾炎组减少（P<0.05, P<0.01）,第21 d时血清肌酐含量低于肾炎组（P<0.05）,光镜和电镜下肾脏病理改变较肾炎组为轻,同时肾皮质内PAF、TXB₂含量少于肾炎组（P<0.01）。结论：川芎嗪治疗实验性肾炎有效,其机理可能与减少肾皮质内PAF、TXB₂等有关。     

银杏内脂B抑制脂多糖诱导的神经炎性细胞毒作用

目的:探讨血小板活化因子受体拮抗剂银杏内脂B(BN52021)抑制细菌脂多糖诱导的脑内炎症反应的神经保护作用。方法:SD大鼠随机分为对照组,模型组和治疗组。Morris水迷宫检测学习和记忆功能,免疫组织化学法检测脑内GFAP阳性星形胶质细胞、OX-42阳性小胶质细胞在颢叶皮质、海马及基底核中的表达。结果:脂多糖(LPS)第4脑室注射使模型动物学习和记忆功能减退,模型组GFAP阳性星形胶质细胞和OX-42阳性小胶质细胞在颞叶皮质、海马及基底核中细胞数量明显增加。BN52021治疗后,对照组和治疗组水迷宫逃避潜伏期、平台象限游泳距离百分比与模型组均有显著性差异;GFAP阳性星形胶质细胞和OX-42阳性小胶质细胞在颞叶皮质、海马及基底核中细胞数量明显减少和阳性染色灰度上升。结论:血小板活化因子受体拮抗剂可抑制LPS诱导的脑内神经炎症,对阿尔茨海默病和艾滋病相关痴呆等以中枢炎症为病理特征的神经退行性变均有治疗作用。     

鞘内注射BN50730抑制SNI大鼠痛敏和脊髓TNF-α的表达

目的： 观察鞘内注射血小板活化因子受体拮抗剂BN50730对坐骨神经分支选择损伤（SNI）神经病理痛大鼠痛阈及脊髓肿瘤坏死因子α（TNF-α）表达的影响,探讨脊髓血小板活化因子(PAF)及其受体参与痛觉信号调节的可能机制。方法: 鞘内置管的Sprague-Dawley大鼠24只随机等分为4组：假手术组,SNI组,SNI+DMSO对照组和SNI+BN50730组,建立SNI疼痛模型,手术后1、3、5、7、10和14 d鞘内给药并测痛阈,第14 d取腰段脊髓免疫组化法和ELISA法检测脊髓TNF-α的表达。结果: SNI神经损伤大鼠机械缩爪阈值明显降低(P<0.05),同侧脊髓背角TNF-α表达增强,ELISA检测脊髓TNF-α含量升高(P<0.05);鞘内应用BN50730降低脊髓TNF-α的表达,同时伴有大鼠痛行为的改善。各组大鼠辐射热缩爪潜伏期无明显差异。结论: BN50730对SNI神经病理性疼痛有治疗作用,TNF-α的表达下调可能与其镇痛机制有关。     

血小板活化因子受体在SAP胰腺组织的变化及银杏苦内酯B的影响

目的探讨血小板活化因子受体(PAF-R)在重症急性胰腺炎(SAP)胰腺组织的动态变化及银杏苦内酯B(BN52021)干预前后的影响。方法Wistar大鼠随机分为阴性对照组(NC组)、SAP模型组(SAP组)、BN52021治疗组(BN组),每组按手术后不同时相点(1、2、3、6、12和24h)分为6小组,分别测定血淀粉酶的变化,并对胰腺组织进行病理学观察和评分,用Westernblot检测胰腺组织PAF-R蛋白质表达的变化。结果BN52021能降低SAP的血淀粉酶和改善其病理变化;SAP组和BN组胰腺组织PAF-R在3h达到高峰(P<0.05),除1h外其余各时相点与NC组有显著性差异(P<0.05);BN组与SAP组之间无显著性差异。结论PAF-R在SAP过程中显著升高,对SAP的病情发展有重要意义,BN52021对PAF-R蛋白表达无显著影响,可能通过竞争性抑制上升的PAF与表达量同时上升的PAF-R相结合,实现它的治疗作用。     

阿托伐他汀对高糖高脂环境下人脐静脉内皮细胞与人肾小球系膜细胞相互作用的影响

 目的 探讨高糖高脂环境下人脐静脉内皮细胞与人肾小球系膜细胞间的相互作用及阿托伐他汀对内皮细胞与系膜细胞相互作用的影响。方法人脐静脉内皮细胞与人肾小球系膜细胞共培养和人肾小球系膜细胞单独培养,分为对照组、甘露醇组、高糖高脂组、BN52021组、阿托伐他汀组。ELISA法检测细胞上清液纤维联接蛋白（Fn）、血小板活化因子（PAF）含量,实时荧光定量检测系膜细胞血小板活化因子受体（PAF-R）mRNA表达。结果 (1)高糖高脂条件下,共培养和单培养Fn、PAF升高（P <0.05）;高糖高脂条件下,共培养Fn、PAF较单培养升高（P <0.05）,BN52021和阿托伐他汀可抑制高糖高脂引起的Fn升高（P <0.05）,阿托伐他汀可抑制高糖高脂引起的PAF升高（P <0.05）;（2）高糖高脂可上调系膜细胞PAF-R mRNA表达（P <0.05）,阿托伐他汀可显著抑制PAF-R mRNA表达上调（P<0.05）。结论 (1) 高糖高脂环境下,系膜细胞和内皮细胞存在异常的相互作用。(2)阿托伐他汀可影响高糖高脂环境下内皮细胞与系膜细胞的相互作用。     

实验性肾炎中肾皮质内血小板活化因子的水平和可能来源

用大鼠复制加速型抗肾小球基底膜肾炎模型，分为４组：肾毒血清对照组，肾毒血清＋眼镜蛇毒因子组，肾毒血清＋羊抗大鼠血小板血清组，肾毒血清＋全身γ线辐射组。各组大鼠在注射兔抗大鼠ＧＢＭ血清后２４ｈ末处死，结果肾毒血清对照组大鼠肾皮质内ＰＡＦ含量显著升高，而耗尽被 体的肾毒血清＋ＣＶＦ组、耗尽血小板的肾毒血清＋ＡＰＳ组和耗尽血白细胞的肾毒血清＋ＴγＩ组大鼠肾皮质内ＰＡＦ含量低于肾毒血清对照组，正常大鼠肾皮     

树鼩血栓性脑缺血时单胺氧化酶活性的变化及银杏内酯B作用机制探讨

目的:揭示血栓性脑缺血时缺血区和血清单胺氧化酶(MAO)活性变化及对血小板活化因子(PAF)受体拮抗剂银杏内酯B(GB)作用机理的探讨。方法:建立光化学诱导树鼠句血栓性脑缺血模型,用酶比色法测量缺血后4、24及72h中心区、半暗区、对侧区及血清的MAO活性,并用双缩脲法测定上述各区的蛋白含量。结果:脑缺血后不同时间缺血中心区MAO活性显著低于假手术组或对侧区,以72h最为明显;此时缺血半暗区和血清MAO活性明显高于假手术组(P＜0.01)。光化学反应后6h舌下静脉一次注射PAF受体拮抗剂GB(5mg·kg^-1)后24h时,半暗区MAO活性明显低于缺血组,而中心区则高于缺血组(P＜0.01)。MAO活性变化与相应区域蛋白含量改变一致(r=0.81,P＜0.05)。结论:脑缺血后中心区、半暗区MAO活性改变是相应区域单胺类递质消长变化的主要原因,MAO活性变化与神经元蛋白质合成能力的改变有关;GB的脑保护作用与其拮抗PAF受体和调节MAO活性而促进递质平衡有关。     

TRAF6在抗β2GPI/β2GPI复合物诱导THP-1细胞表达组织因子时的活化

目的:探讨肿瘤坏死因子受体相关因子6(TRAF6)在抗β2 GPI/β2 GPI复合物诱导单核细胞株THP-1表达组织因子(TF)中的作用.方法:采用一定剂量抗β2 GPI/β2 GPI复合物刺激THP-1细胞一定时间,收集细胞总RNA及总蛋白,实时定量PCR检测细胞TF mRNA水平,发色底物法检测细胞TF活性;RT-qPCR及Western blot分别检测细胞TRAF6mRNA和蛋白表达情况;进一步采用蛋白酶体抑制剂MG-132,观察是否能够干预抗β2 GPI/β2 GPI复合物对细胞的刺激效应.结果:抗β2 GPI/β2 GPI复合物(100 mg/L)能够刺激THP-1细胞表达TF mRNA及活性,与对照相比差异显著(P<0.05);使细胞TRAF6 mRNA和蛋白表达均增加,并显示时间相关性,分别在刺激15 min和30 min时表达至高峰;MG-132(5μmol/L)明显抑制抗β2 GPI/β2 GPI复合物(100 mg/L)对THP-1细胞TRAF6 mRNA和蛋白的刺激效应及TF的诱导表达.结论:抗β2 GPI/β2 GPI复合物诱导THP-1细胞表达TF过程中,TRAF6被激活并发挥重要作用.     

Intraglomerular expressions of IL-1 alpha and platelet-derived growth factor (PDGF-B) mRNA in experimental immune complex-mediated glomerulonephritis.

Acute lung injury frequently develops following haemorrhage, and is characterized by increased proinflammatory cytokine levels and massive neutrophil accumulation in the lung. Blood loss produces rapid increases in tumour necrosis factor-alpha (TNF-alpha) mRNA expression among pulmonary cell populations which precede the development of lung injury. In order to examine the role of TNF-alpha in producing acute inflammatory lung injury, we treated mice following haemorrhage and resuscitation with a TNF antagonist, composed of soluble dimeric human p80 TNF receptor linked to the Fc region of human IgG1 (sTNFR:Fc). Therapy with sTNFR:Fc prevented the post-haemorrhage increases in circulating and pulmonary TNF-alpha levels normally found following blood loss. Administration of sTNFR:Fc also diminished the increase in IL-1 beta, IL-6, TNF-alpha and interferon-gamma (IFN-gamma) mRNA normally found in the lungs following haemorrhage. However, therapy with sTNFR:Fc was not associated with improvement in the histologic parameters of post-haemorrhage lung injury, such as neutrophil infiltration and interstitial oedema. In contrast to the effects of sTNFR:Fc on cytokine mRNA levels among intraparenchymal pulmonary mononuclear cells, such therapy following haemorrhage was associated with increased amounts of mRNA for TNF-alpha among peripheral blood mononuclear cells, as well as increased IFN-gamma titres in serum and bronchoalveolar lavage (BAL) specimens. These results indicate that therapy with sTNFR:Fc in the post-haemorrhage period, although capable of decreasing proinflammatory cytokine expression in the lungs, does not prevent the development of acute lung injury in this setting.     

肿瘤患者血清IL-6、TNF-α、CRP水平与凝血功能的相关性分析

目的通过探讨血清IL-6、TNF-α、CRP水平与凝血因子PT、APTT、D-D、FIB、PLT水平的变化,分析血清炎症因子与凝血指标的关系。方法选取我院2016年4月至2018年4月的肿瘤患者70例,检测血清炎症因子IL-6、TNF-α、CRP水平及凝血因子PT、APTT、D-D、FIB、PLT水平,分析肿瘤患者血清炎症因子与凝血指标的相关性。结果0期肿瘤患者血清症因子IL-6、TNF-α、CRP水平均低于Ⅰ、Ⅱ、Ⅲ期的肿瘤患者,Ⅰ期肿瘤患者血清症因子IL-6、TNF-α、CRP水平均低于Ⅱ、Ⅲ期的肿瘤患者,Ⅱ期瘤患者血清症因子IL-6、TNF-α、CRP水平均低于Ⅲ期的肿瘤患者,差异均有统计学意义(P<0.05)。0期肿瘤患者凝血因子PT、D-D、FIB水平均低于Ⅱ、Ⅲ期的肿瘤患者,0期肿瘤患者凝血因子APTT、PLT均高于Ⅰ、Ⅱ、Ⅲ期的肿瘤患者,Ⅰ期肿瘤患者凝血因子PT、D-D、FIB水平均低于Ⅲ期的肿瘤患者,Ⅰ期肿瘤患者凝血因子PT、D-D、FIB水平均高于Ⅲ期的肿瘤患者,差异均有统计学意义(P<0.05)。血清IL-6水平与凝血因子D-D、FIB水平呈正相关(P<0.05),与凝血因子APTT、PLT水平呈负相关(P<0.05);血清IL-6水平与凝血因子PT无相关(P>0.05)。血清TNF-α、CRP水平与凝血因子PT、D-D、FIB水平均呈正相关(P<0.05),与凝血因子APTT、PLT水平均呈负相关(P<0.05)。结论肿瘤患者成分输血后,凝血功能随着临床分期增加,凝血功能紊反应加剧,且血清IL-6、TNF-α、CRP与凝血功能紊乱有关。     

巨噬细胞细胞毒因子分泌调节的研究

小鼠腹腔巨噬细胞(Mφ)产生细胞毒因子(Mφ—CF),需激活和刺激两个信号。激活信号(A23187)单独不能诱导Mφ—CF,但能促进刺激信号(LPS)的诱导作用。活化的Mφ经LPS作用2小时内开始分泌Mφ—CF,8小时达(?),后逐降。对酵母多糖的吞噬伴有一定程度的Mφ—CF分泌,但吞噬乳胶颗粒则否。数株肿瘤细胞有明显的刺激Mφ—CF分泌的作用。PKc激活剂不能刺激Mφ—CF的产生。     

Purification and characterization of lipopolysaccharide induced TNF-like factor from rabbit serum

A TNF-like factor was purified from lipopolysaccharide (LPS) induced New Zealand white rabbit serum. The TNF-like factor was purified by DEAE-Sephacel, Sephacryl S-200, Mono-Q, CM-affi gel Blue, Superose 12 H/R preparative columns to the specific activity of 4×10⁶ U/mg protein. The purified protein was 45 kDa in its oligomeric form and 22 kDa in its monomeric form. Rabbit TNF-like factor had a pI value of 5.0 and was resistant to trypsin digestion. The TNF-like factor reacted with polyclonal-Ab against human TNF on immunoblot and immunoprecipitation analysis and interacted with human TNF receptors. Taken together, rabbit TNF-like factor might be a high molecular weight form of rabbit TNF.     

Factor V Leiden (R506Q) and risk of venous thromboembolism: a case-control study based on the Spanish population

Resistance to activated protein C (APC) is a frequent cause of thrombophilia. Most patients showing APC-resistance have a G to A mutation at codon 506 of the factor V that converts arginine to glutamine. This mutation is present in populations worldwide with frequencies ranging from 0.01 to 0.05. Genotyping of 150 control individuals from the Spanish population showed that 3.33% of them carried the mutation. Several studies have measured resistance to APC (following a classical functional assay) and have determined the factor V genotype in a number of thrombophilic patients, in an attempt to compare the predictive vaiue of both laboratory methods. To assess the incidence of the factor V mutation among Spanish thrombophilic patients, we genotyped 51 of these. The frequency of mutation carriers rose from 3.33% in the controls to 53% in the patients. We found significant differences for the thrombosis-free survival curves and for the age at the first thrombotic event between patients who carried or did not carry the mutation. Analysis of relatives of 16 patients who carried the factor V mutation suggests the existence of additional genes that modulate the effect of the factor V gene in the development of venous thrombosis among carriers of the G to A mutation.     

血小板第4因子(PF4)抗血管新生的机理初探

研究血小板第４因子抑制血管新生的机制。方法通过研究ＰＦ４与纤维母细胞生长因子－２及其受体间的相互作用来探讨ＰＦ４抑制血管新生的机制。结果：ＦＧＦ２与低亲和力和高亲和力位点的结合受ＰＦ４抑制，这种抑制呈浓度依赖性。５－１０μｇ／ｍｌ的ＰＦ４能量大限度地抑制ＦＧＦ２与低亲和力或高亲和力位点结合。     

中国人动脉血栓性疾病与活化蛋白C裂解FV和FVIII基因位点突变的相关性研究

目的 :检测脑血栓 (CT)和急性心肌梗死 (AMI)患者活化蛋白C(APC)抵抗 (APCR)及APC裂解凝血因子V (FV )和凝血因子VIII(FVIII)肽链的基因位点突变。方法 :用单链构象多态性分析 (SSCP)方法 ,检测 10 2例CT、46例AMI和 10 5例健康人的APC裂解FV肽链 (Arg3 0 6、Arg 5 0 6、Arg 679和结合部位 1865～ 1875 )、裂解FVIII肽链 (Arg3 3 6、Arg 5 62和结合部位 2 0 0 5～ 2 0 18)的基因位点。结果 :对照组和患者组均未检出FV的突变基因。结论 :中国人动脉血栓性疾病患者无APC裂解FV和FVIII肽链的基因突变 ,患者的APC敏感性的降低可能由其他因素所致     

Functional platelet-derived growth factor-beta (PDGF-beta) receptor expressed on early B-lineage precursor cells.

Growth and maturation of B lymphocytes from stem cells require a series of complex processes that are dependent at least in part on growth factors. Uncontrolled expression of receptors from these early growth factors may contribute to a leukaemogenesis of such early B cell progenitors. We show here that early pre-pre-B cells, but not mature B cells, express the PDGF receptor-beta (PDGFR-beta). These receptors contain a protein tyrosine kinase domain which is activated upon ligation with PDGF in pre-pre-B cells. Further, pre-pre-B leukaemia cells seem to express more PDGFR-beta compared with their normal counterparts, suggesting a role for these receptors in growth promotion of leukaemia cells.     

Efficacy,Safety, and Pharmacokinetics of Beroctocog Alfa in Patients Previously Treated for Hemophilia A

Purpose

Beroctocog alfa is a second generation recombinant factor VIII manufactured by removing the B-domain from factor VIII. This prospective clinical trial was conducted to evaluate the efficacy, safety, and pharmacokinetics of beroctocog alfa in patients of ages ≥12 years previously treated for severe hemophilia A.

Materials and Methods

Seventy subjects received beroctocog alfa as an on-demand treatment for acute hemorrhage.

Results

The final hemostatic effect was excellent in 35 subjects (50%) and good in 26 subjects (37.1%). The drug showed an overall efficacy rate of 87.1%. The majority of acute hemorrhages was treated by administering the study drug once (86.2%) or twice (10.0%), and the mean dose administered per single infusion was 28.55±6.53 IU/kg. Ten subjects underwent 12 surgical procedures, and hemostatic efficacy was excellent in seven cases (58.3%) and good in five cases (41.7%), showing a 100% efficacy rate. A total of 52 of 88 subjects (59.0%) experienced 168 adverse events. There were 18 serious adverse events (10.7%) in 11 subjects, and two (mild dyspnea and facial edema) in one subject were related to the study drug. Only one subject formed a de novo factor VIII inhibitor, for an occurrence rate of 1.4% (one-sided 95% upper confidence limit: 3.85%). The final elimination half-life was 13.3 h and 12.6 h at baseline and 6 months after administration, respectively.

Conclusion

Our results suggest that beroctocog alfa is safe and efficacious as either an on-demand treatment for acute hemorrhage or a surgical prophylaxis in patients with hemophilia A.