人消化道癌组织HSP70的检测及其多肽复合物的分离研究

目的 分析人消化道癌组织蛋白表达谱的差异并分离纯化食管癌组织中HSP70-多肽复合物.方法 提取人食管、胃及大肠的癌组织、癌旁组织和正常组织的蛋白质,采用聚丙烯酰胺凝胶电泳方法进行电泳分析;同时以食管癌组织为材料,通过层析柱分离纯化HSP70-多肽复合物.结果 相同器官的不同组织中不同分子量蛋白的种类基本一致,但丰度则表现出差异;同时分离到食管癌细胞的HSP70-多肽复合物.结论 不同分子量的蛋白质在不同组织中表达的丰度不同,可能与组织细胞生命活动状态相关.HSP70-多肽复合物的分离为提取肿瘤HSPT0-多肽复合物提供了具体方法,为深入研究和利用HSP70蛋白奠定了基础.     

血清高敏C反应蛋白、同型半胱氨酸与大脑中动脉斑块内出血的关系北大核心CSCD

目的探讨血清高敏C反应蛋白（hs—CRP）、同型半胱氨酸（Hcy）与大脑中动脉斑块内出血的关系。方法检测63例大脑中动脉主于M1段高度狭窄（狭窄率≥70％）患者（分为症状组与无症状组,斑块内出血组与斑块内无出血组）血清hs—CRP、Hey。所有患者均行常规MRI及高分辨MRI（HR—MRI）检查,分析hs—CRP、Hcy浓度水平与大脑中动脉斑块内出血的相关性。结果症状组37例,无症状组26例。症状组斑块内出血发生率15例（40．5％）高于无症状组3例（11．5％）,两者差异有统计学意义（x^2=6．29,P〈0．05）。症状组[hs—CRP（8．97±3．36）mg／L,Hcy（20．00±3．16）μmol／L3高于无症状组[hs—CRP（5．26±3．12）mg／L,Hcy（12．22±1．88）μmol／L3,两者差异均有统计学意义（t=4．43、11．23,P〈0．001）。斑块内出血组（18／63）hs—CRP（10．53±3．59）mg／L,Hcy（21．70±2．40）μmol／L高于斑块内无出血组（45／63）hs—CRP（6．20±3．02）mg／L,Hcy（11．77±1．69）μmol／L,两者差异均有统计学意义（t=4．87、18．58,P〈0．001）。hs—CRP、Hcy浓度水平与大脑中动脉斑块内出血风险呈正相关（r=0．461、0．519,P〈0．001）。结论血清hs—CRP、Hey水平与大脑中动脉斑块内出血关系较密切,可较好地评估斑块的稳定性;大脑中动脉斑块内出血可能增加同侧大脑中动脉供血区急性脑梗死风险。     

阿托伐他汀对中老年2型糖尿病并高脂血症患者脂联素和瘦素水平的影响北大核心CSCD

目的评价中老年2型糖尿病并高脂血症患者给予阿托伐他汀治疗后血清脂联素、瘦素及其比值的变化。方法选取80例中老年2型糖尿病并高脂血症患者,给予阿托伐他汀10mg／d,连服12周,于用药前及用药12周后分别抽取空腹静脉血,测定空腹血糖（FPG）、糖化血红蛋白（HbA1c）、总胆固醇（TC）、三酰甘油（TG）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）、脂联素（APN）、瘦素（LEP）,并计算脂联素与瘦素比值（A／L）。结果治疗12周前后患者TG、TC、LDL-C、HDL-C分别为[（4．92±1．23）mmol／L和（4．26±1．07）mmol／L,t=11．124,P〈0．001]、[（2．69±1．17）mmol／L和（1．66±1．01）mmol／L,t=24．816,P〈0．001]、[（2．79±1．02）mmol／L和（1．91±0．92）mmol／L,t=21．508,P〈0．001）]、[（1．03±0．30）mmol／L,和（1．33±0．26）mmol／L,t=-12．011,P〈0．001],TG、TC、LDL-C降低,HDL-C升高;APN略有升高,但差异无统计学意义（P=0．064）,LEP水平略有降低（P=0．068）,A／L升高（P〈0．05）。结论中老年2型糖尿病并高脂血症患者经阿托伐他汀治疗后可明显降低TC、TG、LDL-C水平,升高HDL-C水平,并能够升高A／L水平,但对APN、LEP的影响不明届。     

姜黄素抑制棕榈酸诱导的巨噬细胞炎症反应的机制研究

目的探讨姜黄素抑制棕榈酸诱导的巨噬细胞炎症反应的作用机制。方法以0、250、500μmol／L棕榈酸干预小鼠巨噬细胞RAW264．724h,观察细胞形态,逆转录一聚合酶链反应（RT-PCR）检测细胞中受体相互作用蛋白140（RIPl40）、肿瘤坏死因子a（TNF-a）、白细胞介素-6（IL-6）mRNA水平,酶联免疫吸附实验（ELISA）检测上清中TNF-a、IL-6水平;噻唑蓝法（MTr）确定姜黄素作用RAW264．7细胞的最佳时间和浓度;细胞分为姜黄素组和对照组,分别给予20μmol／L姜黄素和二甲基亚砜（DMSO）预处理1h后,均给予500μmol／L棕榈酸作用24h,观察细胞形态并检测细胞中RIPl40、TNF-a、IL-6mRNA水平和上清中TNF-a、IL-6浓度。采用单因素方差分析和t检验对研究数据进行统计检验。结果500μmol／L棕榈酸组RIPl40mRNA表达较0μmol／L组显著升高（3．40±0．51比1．01±0．21,t=7．436,P〈0．01）;0、250、500μmol／L棕榈酸组TNF-amRNA（1．00±0．03、1．79±0．12、2．16±0．13）和蛋白[（197±25）、（371±10）、（485±17）ng／L]、IL-6mRNA（1．00±0．51、2．55±0．15、2．59±0．17）和蛋白[（953±66）、（1081±36）、（1182±18）ng／L]与棕榈酸剂量明显相关（F=99．308、187．049、152．958、26．594,均P〈0．01）;棕榈酸处理后,细胞变圆、皱缩甚至崩解;姜黄素作用RAW264．7细胞的最佳时间为1h,最佳浓度为20ixmol／L;姜黄素组与对照组相比,RIPl40mRNA（1．00±0．05比0．63±0．01,t=一13．79,P〈0．01）、TNF-amRNA（0．64±0．11比1．00±0．07,t=一4．532,P〈0．05）和蛋白[（322±12）比（485±17）ng／L,t=一13．577,P〈0．01]、IL-6mRNA（0．57±0．05比1．00±0．02,t=一14．167,P〈0．01）和蛋白[（241±47）比（1182±18）ng／L,t=一44．810,P〈0．01]均显著降低,而细胞形态正常,细胞问仍有连接融合。结论RIPl40可能参与姜黄素抑制棕榈酸诱导的炎症反应过程。     

热休克蛋白70基因多态性与慢性心力衰竭患者预后的相关性研究

目的:探讨热休克蛋白(HSP)70基因多态性与慢性心力衰竭(CHF)患者预后的相关性。方法:将408例CHF患者纳入研究,根据随访2年内是否发生心血管事件将患者分为事件组与非事件组,采用聚合酶链反应-限制性内切酶分析法检测HSP70-1基因+190G/C、HSP70-2基因+1267A/G和HSP70-hom基因+2437T/C多态性,并收集患者临床资料进行分组及统计学分析。结果:事件组与非事件组患者年龄(t=2.392,P=0.017),脑钠肽(BNP)水平(t=3.922,P=0.000),NYHA分级(χ~2=5.100,P=0.024),左室舒张末期内径(LVEDD)(t=2.872,P=0.004)、右室舒张末期内径(RVEDD)(t=2.232,P=0.026)、左室射血分数(LVEF)超声测值(t=4.338,P=0.000)等参数比较均差异有统计学意义。2组HSP70-2基因+1267A/G位点基因型和等位基因分布频率比较差异均有统计学意义(χ~2=8.480,P=0.014;χ~2=7.643,P=0.006),在隐性基因模型下HSP70-2基因+1267A/G位点基因多态性与CHF患者发生心血管事件相关(χ~2=9.877,P=0.002);HSP70-1基因+190G/C和HSP70-hom基因+2437T/C位点基因型和等位基因分布频率均差异无统计学意义(均P0.05),且在显性基因模型和隐性基因模型下两位点基因多态性均与CHF患者发生心血管事件无关(均P0.05)。Logistics多因素回归分析结果显示HSP70-2基因+1267A/G位点GG基因型是CHF患者发生心血管事件的危险因素(OR=1.362;95%CI 1.046~2.701;P=0.003)。结论:HSP70-2基因+1267A/G多态性与CHF患者预后相关,GG基因型可增加CHF患者发生心血管事件的风险。HSP70-1基因+190G/C和HSP70-hom基因+2437T/C多态性可能与CHF患者预后关联不大。     

呼吸道合胞病毒M2:81-95与热休克蛋白70L1融合蛋白的制备及其免疫原性

目的 构建呼吸道合胞病毒(RSV)CTL表位M2:81-95与热休克蛋白(HSP)70L1融合蛋白的重组质粒,原核表达后初步研究其免疫原性.方法 从SMMC7721细胞中克隆HSP70L1基因.PCR扩增M2:81-95基因片段,鉴定后构建质粒pET-HSP70L1-M2:81-95(pET-HSP70L1-M2).经PCR和酶切鉴定,转化E.coli BL21(DE3).用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达HSP70L1-M2:81-95(HSP70L1-M2),Ni-螫合亲合层析纯化,梯度透析复性.用该蛋白免疫10只BAlB/c小鼠,ELISA榆测IgG抗体及其亚型.噻唑蓝(MTT)法榆测CTL杀伤活性.结果 成功构建重组质粒pET-HSP70L1-M2,并在E.coli中表达重组蛋白HSP70L1-M2,该蛋白诱导RSV及表位特异性的CTL活性,还诱导蛋白抗原特异性的IgG,为4.87±0.35、IgGl为5.53±0.28、IgG2a为4.40±0.21.且IgG1/IgG2a(1.26)的比例平衡,与PBS对照组的IgG,为0.33±0.17、IgG1为0.51±0.21、IgG2a为0比较,差异有统计学意义(t=3.512,3.681,5.856;P<0.05).结论 成功构建原核表达质粒pET-HSP70L1-M2,斤表达纯化获得重组蛋白,免疫小鼠后诱导RSV及表位特异性的CTL活性和辅助性T淋巴细胞(Th)1/Th2混合型应答.     

白果内酯在鱼藤酮处理的PC12细胞中抑制α-共核蛋白蛋白寡聚体形成的实验研究

目的研究白果内酯对鱼藤酮诱导的帕金森病细胞模型是否具有保护作用,对α-共核蛋白（α-synuclein）的表达及聚集是否有影响。方法使用鱼藤酮对嗜铬细胞瘤株PCl2细胞进行处理,建立叶synuclein蛋白高表达的帕金森病细胞模型,白果内酯进行干预,采用四甲基偶氮唑盐法（MTT法）检测细胞活性,流式细胞术检测细胞凋亡,Western blotting法检测α-synuclein蛋白的表达。数据以均数±标准差（面±5）表示,应用SPSS16．0统计软件,多组间比较采用单因素方差分析,两组间独立样本的比较采用LSD—t检验,P〈0．05认为差异有统计学意义。结果1．6μmoL／L鱼藤酮组细胞活性显著低于对照组（t=17．422,P〈0．01）,细胞凋亡率和α--synuclein蛋白量高于对照组（t=9．141,t=8．392;P均〈0．01）;10μmol／L及50／μmol／L白果内酯组细胞活性均高于鱼藤酮组（t=4．257,t=6．501;P均〈0．01）,且高剂量者细胞活性更优（t=2．933,P=0．043）;10μmol／L及50μmoL／L白果内酯组细胞凋亡率较鱼藤酮组低（t=4．482,t=4．488,P均〈0．01）,α-synuclein蛋白的表达量低于鱼藤酮组（t=8．349,t：9．028,P均〈0．01）,但两剂量间二者均无统计学差异（￡=0．831,P=0．45;t=2．178,P=0．095）;实验中所显影的α-synuclein蛋白分子量在57kD左右。结论白果内酯对鱼藤酮诱导的PCI2细胞损伤有保护作用,该保护作用可能通过抑制仅,synuclein蛋白寡聚物的形成来实现。     

椎动脉扭曲与后循环缺血的相关性

目的：探讨椎动脉扭曲与后循环缺血(posterior circulation ischemia, PCI)的相关性。方法纳入年龄≥50岁的 PCI 患者和同期无脑缺血表现的对照者。所有患者均行 CT 血管造影,观察颈部椎动脉扭曲情况并进行评级,分析影响 PCI 的相关危险因素。结果共纳入112例PCI 患者和90例对照者。单变量分析显示,PCI 组高血压(80．36%对54．44%;χ2=15．613,P <0．001)、吸烟(35．71%对18．89%;χ2=6．974,P =0．008)、饮酒(25．89%对10．00%;χ2=8．253,P =0．004)、后循环血管狭窄(54．46%对24．44%;χ2=18．578,P <0．001)和椎动脉扭曲(71．43%对48．89%;χ2=10．695, P =0．001)的患者比例以及总胆固醇[(4．96±1．26)mmol/L 对(4．61±1．04)mmol/L;t =-2．110, P =0．036]、低密度脂蛋白胆固醇[(3．02±0．90)mmol/L 对(2．69±0．78)mmol/L;t =-2．671,P =0．008]和纤维蛋白原[(3．67±1．69)mg/L 对(3．25±0．97)mg/L;t =-2．002,P =0．047]水平均显著高于对照组。 PCI 组双侧椎动脉均扭曲的比例显著高于对照组(30．36%对12．22%;χ2=9．478,P =0．002)。 PCI 组3级椎动脉扭曲的比例显著高于对照组(43．75%对26．67%;χ2=6．310,P =0．012)。多变量 logistic 回归分析显示,吸烟[优势比(odds ratio, OR)2．339,95%可信区间(confidence interval, CI)1．037~5．278;P =0．041]、低密度脂蛋白胆固醇(OR 1．580,95% CI 1．050~2．377;P =0．028)、高血压(OR 2．631,95% CI 1．237~5．596;P =0．012)、后循环血管狭窄(OR 3．419,95% CI 1．638~7．134;P =0．001)和椎动脉扭曲(OR 2．413,95% CI 1．212~4．803;P =0．012)是 PCI 的独立危险因素。结论椎动脉扭曲是中老年 PCI 的独立危险因素。     

血清尿酸、胆红素水平与急性缺血性卒中患者近期转归的相关性

目的：探讨基线尿酸水平及胆红素水平与急性缺血性卒中患者短期转归的关系。方法收集缺血性卒中患者的临床资料,包括入院时血清尿酸和胆红素水平、美国国立卫生研究院卒中量表(National Institutes of Health Stroke Scale, NIHSS)评分、出院时或第14天时改良 Rankin 量表(modified Rankin Scale, mRS)评分(0~2分定义为转归良好,>2分定义为转归不良)。结果共纳入急性缺血性卒中患者162例,转归良好组114例,转归不良组48例。2组之间糖尿病(51．75%对75．00%;χ2=7．526,P =0．006)、既往卒中或短暂性脑缺血发作( transient ischemic attack, TIA)史(18．42%对50．00%;χ2=17．790, P <0．001)的患者构成比以及基线舒张压[(87．061±12．245)mmHg 对(82．375±10．949)mmHg,1 mmHg =0．133 kPa;t =2．293,P =0．023]、高密度脂蛋白胆固醇[(1．604±0．299)mmol/L 对(1．265±0．206)mmol/L; t =3．227, P =0．002]、空腹血糖[(2．875±0．438)mmol/L 对(8．160±0．592)mmol/L; t =-4．761, P <0．001)]、尿酸[(289．365±77．168)μmol/L 对(248．206±66．206)μmol/L; t =3．111, P =0．002]、总胆红素[(14．673±2．213)μmol/L 对(10．395±2．714)μmol/L; t =3．779, P <0．001]、直接胆红素[(6．036±1．392)μmol/L 对(4．956±1．379)μmol/L; t =2．088, P =0．038]、间接胆红素[(8．634±2．307)μmol/L 对(5．439±1．223)μmol/L;t =4．219,P <0．001]水平存在显著差异。多变量 logistic回归分析显示,既往卒中或 TIA 史[优势比(odds ratio, OR)3．751,95%可信区间(confidence interval, CI)1．395~10．091;P =0．009]和基线 NIHSS 评分(OR 2．723,95% CI 1．093~6．783;P =0．031)是缺血性卒中转归不良的独立危险因素,而尿酸(OR 0．357,95% CI 0．141~0．900;P =0．029)、高密度脂蛋白胆固醇(OR 0．262,95% CI 0．079~0．870;P =0．029)和间接胆红素(OR 0．117,95% CI 0．025~0．539;P =0．006)与转归良好独立相关。结论基线尿酸和间接胆红素水平增高是急性缺血性卒中患者转归良好的有利因素。     

叉头状转录因子O1对游离脂肪酸诱导的人肝癌细胞株HepG-2胰岛素抵抗和脂质堆积作用的研究

目的研究叉头状转录因子O1（FoxO1）对游离脂肪酸介导的人肝癌细胞株（HepG-2）胰岛素抵抗和脂质堆积的作用以及对固醇调节元件结合蛋白-1c（SREBP-1c）mRNA和蛋白表达的影响。方法将HepG-2细胞培养后用普通培养基培养为对照组,用含5．0×10μmol／L软脂酸的培养基诱导为软脂酸组,诱导后转染空白质粒为空白质粒组,诱导后转染FoxO1siRNA质粒载体为FoxO1siRNA载体组。运用实时定量聚合酶链反应（RT—PCR）方法检测FoxO1mRNA表达,噻唑蓝（M33＂）比色法检测细胞增殖,葡萄糖氧化酶法检测培养基中葡萄糖消耗量,油红O染色观察细胞脂质堆积;RT—PCR和Westernblot技术分别检测SREBP-1c mRNA表达量以及蛋白的表达量。各组间均值比较采用单因素方差分析,样本间比较采用t检验。结果软脂酸组较对照组细胞葡萄糖消耗量减少（1．174-0．56vsVS4．31±0．21,t=10．587,P〈0．01）、细胞中的脂质堆积增多、FoxO1mRNA升高（0．784-0．10vs0．51±0．12,t=3．629,P〈0．05）、SREBP-1cmRNA升高（0．71±0．17vs0．25±0．08,t=6．290,P〈0．05）、SREBP-1c蛋白升高（0．694-0．10vs0．41±0．07,t=4．797,P〈0．01）。转染FoxO1siRNA质粒载体后葡萄糖消耗量较软脂酸组增加（2．26±0．41vs1．17±0．56,t=3．144,P〈0．05）,FoxO1mRNA、SREBP-1cmRNA、SREBP-1c蛋白的表达较软脂酸组均减少且接近于对照组（分别为0．38±0．06vs0．784-0．10,t=7．164,P〈0．01;0．45±0．13vs0．71±0．17,t=2．479,P〈0．05;0．41±0．06vs0．694-0．10,t=4．797,P〈0．01）,细胞中的脂质堆积也较软脂酸组减少。结论抑制FoxO1的表达,可改善游离脂肪酸诱导的细胞胰岛素抵抗、减少肝脏细胞内脂肪变性,其机制可能是通过下调SREBP-1c的表达。     

冠心病患者HSP70-2+1267A/G多态性与高敏C反应蛋白水平的相关性研究

冠心病（coronary artery disease,CAD）的病理基础是动脉粥样硬化,炎症是动脉粥样硬化的特征之一;研究发现炎症在冠状动脉粥样硬化斑块的发生、发展和粥样斑块的破裂过程中起重要作用,动脉粥样硬化被认为是一种慢性炎症的过程。热休克蛋白70-2（Hot Shock Protein,HSP）是HSP家族中重要的一员,     

Purification of heat shock protein 70-associated tumor peptides and their antitumor immunity to hepatoma in mice

AIM:To purify the heat shock protein (HSP) 70-associated tumor peptides and to observe its non-MHC-I molecule restrictive antitumor effect.METHODS:By ConA-sepharose affinity chromatography,ADP-agarose affinity chromatography, and DEAE anion exchange chromatography, we were able to purify HSP70-associated peptides from mouse hepatoma (HCaF) cells treated in heat shock at 42℃. Specific active immunization and adoptive cellular immunization assay were adopted to observe the immunoprotective effect elicited by HSP70-associated peptide complexes isolated from HcaF.RESULTS: The finally purified HSP-associated peptides had a very high purity and specificity found by SDS-PAGE and Western blot. Mice immunized with HSP70-associated peptide complexes purified from HCaF cells were protected from HCaF living cell challenge. This effect was dose dependent.Adoptive immunization of immune spleen cells of mice immunized with HSP70-associated peptide complexes could elicit immunity against HCaF challenge, and the tumor-free mice could resist repeated challenges. This effect could be continuously enhanced by repeated challenge with HCaF living cells. The tumor-free mice could tolerate the challenge for as high as 1&#215;10^7 HCaF cells. The mice immunized once with spleen cells pulsed with HSP70-associated peptide complexes in vitro could also result in a certain adoptive immunity against HCaF.CONCLUSION:High purity and specificity of HSP70-associated peptides could be achieved from tumor cells by the low-pressure affinity chromatography method used in this study. HSP70-associated peptide complexes derived from the HCaF can elicit non-MHC-I molecule restrictive immunoprotective effect against HCaF.This effect can be transferred by adoptive immunization to mice and enhanced by repeated challenge with HCaF live cells.     

The major histocompatibility complex region marked by HSP70-1 and HSP70- 2 variants is associated with clozapine-induced agranulocytosis in two different ethnic groups

Genes of the major histocompatibility complex (MHC) have been associated with susceptibility to drug-induced adverse reactions. We previously found that clozapine-induced agranulocytosis (CA) is associated with the HLA-DRB1*0402, DRB4*0101, DQB1*0302, DQA1*0301 haplotype in Ashkenazi Jewish patients and with the HLA-DRB1*1601, DRB5*02, DQB1*0502, DQA1*0102 haplotype in non-Jewish patients. In the present study, we tested the hypothesis that the variants of the heat- shock protein 70 (HSP-70) encoded by the HSP-70 loci located within the MHC region and known to be involved in apoptosis and regulation of cell proliferation could play an important role in molecular mechanisms of CA. First, we analyzed HSP70-2 polymorphism in risk-associated haplotypes from HLA homozygous cells and normal individuals and confirmed that the HSP70-2 9-kb variant was associated invariably with DR4 (HLA-DRB1*0402, DQB1*0302) and DR2 (HLA-DRB1*01601, DQB1*0502, DQA1*0102 and HLA-DRB1*1501, DQB1*0602) haplotypes, which were the haplotypes found increased in Jewish and non-Jewish patients with CA, respectively. The 9.0-kb variant was also found to be associated with HLA-B44, DRB1*0401 and HLA-B44, DRB1*07 haplotypes. Second, in patients with CA (12 Ashkenazi Jewish and 20 non-Jewish patients), HSP70-1 A and HSP70-2 9.0-kb variants were associated with the MHC haplotypes found by us to be markers of susceptibility to CA. The clozapine-treated control group had an excess number of HSP70-1 C and HSP70-2 8.5-kb variants, consistent with genetic resistance to CA associated with those variants. This finding supports our hypothesis that a dominant gene within the MHC region (marked by HSP70-1 and HSP70-2), but not necessarily HLA, is associated with CA in two different ethnic groups.     

Peripheral blood mononuclear cell proliferation to heat shock protein-70 derived from autologous lung carcinoma

In animals, tumor-derived heat shock proteins (HSP) induce immune-mediated protection against autologous cancer. We investigated whether HSP-70 derived from human lung carcinoma are also complexed to tumor-specific antigens. Peripheral blood mononuclear cells collected 10 days after surgery from patients with lung cancer were stimulated with HSP-70 purified from autologous and heterologous tumors. The stimulation index (SI) obtained when stimulating cells with autologous tumor-derived HSP-70 averaged 3.07 +/- 0.75 in patients with lung cancer and 1.57 +/- 0.33 in control subjects (p < 0.001 by analysis of variance). No significant stimulation was observed with HSP-70 derived either from the majority of heterologous tumors or from autologous tumor-free lung tissue. SI decreased from 3.59 +/- 0.65 to 1.65 +/- 0.38 in six patients tested again 3 months after surgery (p = 0.02 by Wilcoxon test for paired data). HSP-70 derived from lung carcinoma are shown to be associated with T cell antigens. The T cell reactivity appears transient and restricted to antigens complexed to HSP-70 derived from autologous tumors only. This suggests that the antigenicity of human lung tumors is unique, which may be crucial for the design of new vaccines.     

The magnitude of the immune response to heat shock protein-65 following BCG immunisation is associated with the extent of experimental atherosclerosis

Several studies have reported associations between coronary heart disease (CHD) and infection. Recent studies have implicated immune responses to heat shock protein(s) (HSP) as a contributary factor. Using an immunisation model, we have assessed the relationship between the immune responses to HSP and subsequent atherosclerosis. Rabbits were immunised with bacillus Calmette-Guerin (BCG) vaccine (n=10) or saline (n=10) and subsequently fed a 0.25-1.0% cholesterol diet for 10 weeks. Plasma levels of IgG specific for mycobacterial antigen A60 and human HSP-60, but not for human HSP-70, rose following BCG immunisation, reaching a peak after 8 weeks. The percentage aortic area covered by atherosclerotic plaque was greater in animals immunised with BCG (30.5+/-3.8) compared to saline treated animals (16.4+/-2.6) (P<0.05). Furthermore, the individual titres of anti-HSP-60 in the BCG-immunised animal antibodies at week 8 (prior to starting the cholesterol diet) correlated with the percentage aortic area covered by plaque after 18 weeks (R2=0.72; P<0.05). No correlation was found between anti-A60 antibody titres and plaque area. Antiserum from BCG-immunised, but not control, animals stained heat-shocked endothelial cells. These data suggest that immune responses to HSP may be implicated in the relationship between specific infections and CHD.     

Effect of HSP70 induced by warm ischemia to the liver on liver function after partial hepatectomy

BACKGROUND/AIMS: The purpose of this study was to determine if induction of HSP70 (heat shock protein 70), a stress protein which plays a cytoprotective role in response to various stimuli, protects hepatocytes from damage caused by partial hepatectomy and, if so, to elucidate the mechanism of such protection. METHODOLOGY: One hundred and eight male F344 rats weighing 190-220 g were randomly assigned to two groups with or without the presence of preconditioning. Fifteen-minute warm ischemia was applied to the liver of rats to induce HSP70, and 70% hepatectomy was performed 48 hours after the induction of HSP70 (ischemia group; n = 72). The rats in the nonischemia group did not undergo 15-min warm ischemia prior to 70% hepatectomy (nonischemia group; n = 36). Six rats, selected randomly from each group, were sacrificed at each measurement point to obtain blood and liver tissue samples. The levels of HSP70 in the liver, serum nitric oxide, levels of catalase and superoxide dismutase activity in the liver as antioxidative enzymes, and levels of Bcl-xL and Bax proteins and caspase-3-like activity in the liver as indices of apoptosis, were measured. RESULTS: The mean +/- SD level of HSP70 in the ischemia group (100 +/- 42 arbitrary unit (au)) was significantly higher than that of the nonischemia group (2 +/- 0.7 au) immediately before hepatectomy (P < 0.05). The ischemic preconditioning attenuated the liver damage caused by the subsequent partial hepatectomy. The levels of superoxide dismutase and catalase activity, serum nitric oxide level, and Bax protein level of the ischemia and nonischemia groups showed no significant differences after the partial hepatectomy. In contrast, the mean +/- SD level of Bcl-xL in the liver of the ischemia group (261 +/- 52 au) was significantly higher than that in the nonischemia group (114 +/- 33 au) 12 hours after the hepatectomy (P < 0.01). Furthermore, the mean +/- SD level of caspase-3-like activity in the liver of the ischemia group (18.1 +/- 4.6 au) was significantly lower than that of the nonischemia group (26.0 +/- 4.8 au) at 12 hours after the hepatectomy (P < 0.05). CONCLUSIONS: HSP70 induced by ischemic preconditioning prior to the partial hepatectomy was considered to protect the liver itself. In addition, the induced HSP70 may affect the Bcl-xL level after partial hepatectomy. Therefore, Bcl-xL seems to be involved in the reduction of liver damage after partial hepatectomy along with HSP.     

The role of heat shock protein 27 expression in hepatocellular carcinoma in Japan: special reference to the difference between hepatitis B and C.

BACKGROUND: A recent report showed that heat shock protein (HSP)-27 expression was related to histological grade and survival of patients with hepatocellular carcinoma (HCC). AIMS: The aim of this study was to examine the effect of expression of HSP-27 on clinicopathological variables in Japanese patients with HCC. METHODS: An immunohistochemical study for HSP-27 was performed on 60 HCC cases using a monoclonal anti-HSP-27 antibody. We divided 60 patients into two groups, patients with a low expression of HSP-27 (n = 34) and those with a high expression of HSP-27 (n = 26). Forty patients tested positive for the hepatitis C virus (HCV) antibody and 20 tested positive for the hepatitis B surface antigen. RESULTS: There appeared to be no relationship between HSP expression and clinicopathologic factors and no differences were observed between the high expression group and the low expression group. In the hepatitis B virus (HBV) group (n = 20), HSP-27 expression correlated significantly with prognosis, disease-free survival (DFS) and overall survival. High expression was significantly associated with poor prognosis in the HBV group. In contrast, patients with a high expression tended to have a good prognosis in the HCV group (n = 40): DFS and overall survival. CONCLUSIONS: This study showed the possibility that HSP-27 plays different roles in HBV- and HCV-associated HCCs.