HSP27、70、90α和105在Barrett's 食管中的表达及其相互关系

目的 研究热休克蛋白27、70、90α、105(HSP27、70、90α、105)在Barrett's 食管(Barrett's esophagus, BE)和食管炎伴贲门腺或胃底腺化生组织中的表达及相互关系,探讨其在BE发病中的作用.方法 应用免疫组织化学法(LDP)检测22例BE和38例食管炎伴贲门腺或胃底腺化生组织中HSP27、70、90α和105的表达,并比较其表达的相关性.结果 HSP70和HSP90α在BE中的阳性表达率分别为31.82%和9.09%,明显低于食管炎伴贲门腺或胃底腺化生组织中的表达(47.37%、42.11%)(P=0.001和P=0.000);HSP105在BE中的阳性表达率为100%,明显高于食管炎伴贲门腺或胃底腺化生组织中的表达(63.16%)(P=0.000);HSP27在BE中的阳性表达率为77.27%,与食管炎伴贲门腺或胃底腺化生组织中的表达率相比无明显差异(50.00%)(P=0.422).BE中HSP70和HSP90α的表达存在强正相关(r=1, P=0.000),HSP70、HSP90α的表达均与HSP105的表达存在强负相关(r=-1, P=0.000).结论 BE上皮中的肠化细胞凋亡和增殖失调,HSP70、HSP90α和HSP105在BE发生过程中起重要作用,可能成为预测BE发生的重要指标.     

三七总皂甙与参麦注射液对大鼠脑出血后脑水肿、行为学变化的影响

目的 比较观察三七总皂苷及参麦注射液对脑出血大鼠脑组织损伤和脑水肿的作用.方法 采用胶原酶与肝素复合法造成大鼠脑出血模型,造模后3 h,各组按试验设计给药,在相应时间点先对大鼠进行神经行为学评分,测定脑组织含水量、细胞形态学的变化,TNF-α及HSP70表达.结果 三七、参麦均能够改善神经功能缺失症状,在第4天与模型组比较差异显著(P＜0.05).三七及参麦组能够减轻脑组织水肿,于第4天、第7天改善明显,与模型组比较差异显著(P＜0.05),与甘油作用相似,但起效较慢.三七和参麦均能够使大鼠脑出血后TNF-α表达降低、HSP70表达升高,以4 d、7 d与模型组比较有显著性差异(P＜0.05).结论 三七和参麦可以改善脑出血后神经功能缺陷症状;能减轻脑出血后脑水肿;可以抑制脑组织TNF-α的表达,增加HSP70表达,疗效优于甘油;综合各指标分析和比较,三七的作用较参麦好.     

艾灸预处理对脑缺血大鼠HSP70蛋白及HSP70mRNA表达的影响

目的探讨艾灸预处理的预防性脑保护作用机制。方法健康Wistar雄性大鼠随机分为正常对照组、假手术组、脑缺血组、脑缺血预处理组、艾灸预处理组。正常组正常饲养;假手术组仅暴露四条血管,不发生脑缺血;脑缺血组全脑缺血10min;脑缺血预处理组先缺血预处理3 min,再灌注24 h后再次全脑缺血10 min。艾灸预处理组先给予艾灸预处理7 d,再行全脑缺血10 min。各组分别于手术后24、48、72 h取脑,采用免疫组化法、原位杂交法检测海马CA1区HSP70蛋白及HSP70mRNA的表达。结果脑缺血组24 h HSP70蛋白表达达到高峰,72 h蛋白含量显著降低(P<0.01)。脑缺血预处理组、艾灸预处理组可见较多HSP70蛋白表达,较缺血组明显增高(P<0.01)。艾灸预处理组术后24、48 h HSP70蛋白的表达较缺血预处理组明显增高(P<0.01、P<0.05);72 h HSP70蛋白的表达明显降低,与缺血预处理组比较具有显著性差异(P<0.01)。结论艾灸预处理能诱导脑缺血大鼠海马CA1区HSP70 mRNA、HSP70蛋白的表达。其预防性脑保护作用可通过促进HSP70蛋白及HSP70mRNA表达高峰...     

亚低温对大鼠脑缺血再灌注损伤后HSP70 、TNF-α、NF-κB表达及损伤神经细胞凋亡的影响

目的：观察亚低温对大鼠脑缺血再灌注损伤后HSP70（热休克蛋白70），TNF－α（肿瘤坏死因子－α），NF－κB（核蛋白因子－κB）表达及损伤神经细胞凋亡的影响，方法：采用大鼠局灶性脑缺血再灌注损伤模型，大脑中动脉阻塞2h，再灌注损伤10h，用免疫组织化学法和原位缺口末端标记（TUNEL）法分别检测假手术组，对照组和亚低温组HSP70，TNF－α，NF－κB表达水平和凋亡细胞百分率。结果：亚低温组HSP70表达水平对照组显著升高（P＜0．05），而TNF－α，NF－κB表达水平和凋亡细胞百分率明显低于对照组（P＜0．05），结论：亚低温下调大鼠脑缺血再灌注损伤后TNF－α，NF－κB表达水平和上调HSP70表达水平可能与其抗损伤神经细胞凋亡作用有关。     

白花丹参叶制剂对局灶性脑梗死HSP70及细胞凋亡的影响

目的观察热休克蛋白70(HSP70)、神经细胞凋亡在大鼠局灶性脑梗死后不同时程的表达,探讨二者的关系及白花丹参叶制剂对其的影响.方法采用光化学法诱导大鼠局灶性脑梗死模型,HSP70免疫组织化学染色,并用TdT-介导duTP-生物缺口末端标记(TUNEL)方法检测凋亡细胞.结果免疫组化显示白花丹参叶制剂预处理组与单纯局灶性脑梗死组各时间点相比阳性反应均有明显的增多(P＜0.05).TUNEL结果显示,白花丹参叶制剂预处理组各时间点凋亡细胞均有统计学意义.结论白花丹参对脑梗死具有保护作用.     

鹦鹉热衣原体HSP60基因克隆及其表达产物抗原性的实验研究

目的验证新获得的鹦鹉热衣原体HSP60基因在E.coli中高效表达的产物是否具有足够的抗原活性,为进一步研究其在鹦鹉热衣原体感染检测上的应用奠定基础。方法根据GenBank鹦鹉热衣原体Hsp60基因序列X51404,设计一对特异性引物,扩增了鹦鹉热衣原体HSP60全长基因,克隆入pET32a(+)载体,转化大肠杆菌BL21(DE3)后,利用IPTG诱导获得高效表达。结果扩增了鹦鹉热衣原体HSP60全长基因,表达产物分子量约为68kD,经Western blotting和胶体金免疫层析技术检测,表明表达产物具有较好的抗原性。结论扩增了鹦鹉热衣原体HSP60全长基因,并对其表达产物的抗原性进行免疫印记分析和免疫层析初步检测,表明具有较好的抗原性,为进一步研究其在鹦鹉热衣原体感染检测上的意义打下了基础。     

HSP70反义寡脱氧核苷酸促进MRC-5细胞衰老的研究

目的 采用反义寡核苷酸技术了解HSP70与细胞衰老之间的关系.方法 设正义组、反义组,在转染后12,24 h,3,5 d,分别用流式细胞技术和免疫荧光技术检测HSP70蛋白水平、凋亡细胞比例和细胞周期.在转染后第2、4、6天,分别收集正义组、反义组和空白对照组细胞爬片进行SA-β-Gal染色.结果 各时间点反义组蛋白表达均低于正义组（P＜0.05）,凋亡细胞比例均高于正义组（P＜0.05）.随观察期延长,处于G1期的细胞数逐渐增多,但反义组均高于正义组（P＜0.05）.相同时间点反义组SA-β-Gal染色阳性细胞（蓝染细胞）数均较正义组和空白对照组多,随观察期延长,各组蓝染细胞均增多.结论 HSP70反义寡脱氧核苷酸下调MRC-5细胞HSP70的表达,使细胞出现G1期阻滞,促进细胞衰老.     

左归丸对老年性痴呆模型鼠脑神经元HSP70及超微结构的影响

目的 探讨左归丸对老年性痴呆(Alzheimer病,AD)动物模型HSPT0及超微结构的影响.方法 模型组以D-半乳糖腹腔注射合并A13注射海马制备AD动物模型,各治疗组分别在造模的同时灌胃左归丸和抗脑衰胶囊、哈伯因.采用生化法检测各组大鼠脑匀浆AchE,运用RT-PCR测定各组海马HSP70的表达并用透射电镜观察各组海马神经元超微结构的变化.结果 左归丸能明显抑制模型鼠脑组织中AchE活性,上调HSP70的表达,抑制细胞凋亡.超微结构观察发现模型组海马神经元显现凋亡早期的形态特点,各治疗组和对照组对凋亡皆有抑制作用.结论 左归丸对痴呆鼠海马神经元细胞凋亡有抑制作用,其机制可能与其抑制模型鼠脑组织中AchE活性、上调HSP70的表达和改善神经元细胞损害有关.     

胰岛素对全脑缺血再灌注大鼠脑HSP70表达的作用

目的探讨胰岛素对脑缺血再灌注损伤的中枢保护机制。方法通过胸部挤压建立心脏骤停后完全性全脑缺血再灌注模型,观察HSP70在脑组织的表达情况。结果与对照组相比,胰岛素并不能显著上调缺血大脑的HSP70表达。结论胰岛素对全脑缺血大鼠大脑的HSP70表达没有显著影响。     

Tanespimycin with bortezomib: activity in relapsed/refractory patients with multiple myeloma

Tanespimycin (17‐allylamino‐17‐demethoxygeldanamycin, 17‐AAG) disrupts heat shock protein 90 (HSP90), a key molecular chaperone for signal transduction proteins critical to myeloma growth, survival and drug resistance. In previous studies, tanespimycin monotherapy was well tolerated and active in heavily pretreated patients with relapsed/refractory multiple myeloma (MM). Preclinical data have shown antitumour synergy between tanespimycin and bortezomib, with more pronounced intracellular accumulation of ubiquitinated proteins than either drug alone, an effect attributed to the synergistic suppression of chymotryptic activity in the 20S proteasome. HSP70 induction has been observed in all Phase 1 tanespimycin studies in which it has been measured, with several separate reports of HSP70 overexpression protecting against peripheral nerve injury. In this Phase 2, open‐label multicentre study, we compared 1·3 mg/m² bortezomib + three doses of tanespimycin: 50, 175 and 340 mg/m² in heavily pretreated patients with relapsed and refractory MM and measured HSP70 expression and proteasome activity levels in plasma of treated patients. The study was closed prematurely for resource‐based reasons, precluding dose comparison. Nonetheless, antitumour activity was observed, with promising response rates and promising severity of peripheral neuropathy.     

沙鼠前脑缺血再灌注后脑组织HSP20和EPO的表达变化及相关性

目的 观察沙鼠前脑短暂性缺血再灌注后,HSP20与EPO在脑组织中的表达,探讨二者的相关性和保护机制,并评估它们的临床应用前景.方法 45只健康雄性蒙古沙鼠随机分为三组:正常对照组、假手术组和脑缺血再灌注组,其中假手术组和脑缺血再灌注组又分为4个亚组:6 h组,1 d组,3 d组和7 d组.予以夹闭双侧颈总动脉并再通造成脑缺血再灌注模型.HE染色观察沙鼠前脑组织的病理变化.免疫组织化学染色观察HSP20和EPO的表达情况.结果 HE染色:正常对照组和各假手术组的沙鼠前脑组织形态结构没有明显的变化;脑缺血再灌注组根据时间点的不同可见胶质细胞肥大、增生,细胞和细胞间质水肿,神经元坏死.免疫组化染色:与对照组及假手术组相比,缺血再灌注损伤后HSP20及EPO的表达均上调,于第三天达到高峰,后开始下降至7 d时仍高于正常组和假手术组(P<0.05),且两者的表达趋势相同呈正相关(r=0.777,P<0.05).结论 缺血再灌注的沙鼠脑组织中HSP20及EPO表达均升高并可能减轻缺血再灌注所导致的脑损伤.     

热休克蛋白70对内毒素诱导的肝脏NF-κB信号转导通路的影响

目的探讨热休克蛋白70对内毒素（LPS）诱导的肝脏NF-κB信号转导通路的影响。方法雄性昆明种小鼠70只随机分为3组,对照组（n=10）、LPS诱导组（n=30）、亚砷酸钠（SA）预处理组（n=30）,分别于处理后0.5 h、1.5 h和6 h在麻醉下开腹取肝脏,Western blot方法检测肝脏NF-κB P65、NF-κB P50和IκB的表达。结果 SA预处理可抑制NF-κB P65和NF-κB P50蛋白的表达,增强肝脏IκB蛋白的表达。结论亚砷酸钠（SA）刺激机体产生HSP70,可通过影响肝脏信号转导,从而抑制相关炎症通路,减轻LPS所致的急性肝损伤。     

鸡源性肠炎沙门氏菌的流行病学调查

目的与方法通过对一株分离自患病蛋鸡的沙门氏菌进行了生化鉴定、血清学鉴定、分子生物学检测和攻毒试验,确定了我国鸡群中存在肠炎沙门氏菌的感染;通过血清学试验,对我国部分鸡群进行了血清流行病学调查。结果结果表明,从临床分离的沙门氏菌与肠炎沙门氏菌的微生物学和生物化学特征相符合,通过血清型鉴定、PCR检测、DNA测序和分析证明分离的肠炎沙门氏菌株与标准株肠炎沙门氏菌相符合;首次全国范围血清流行病学调查,从全国采集603份蛋鸡血清中发现9份阳性样本,利用肠炎沙门氏菌阳性血清检测从当地禽肉、蛋样本中分离的8株可疑沙门氏菌,未检测到肠炎沙门氏菌。结论本研究结果显示,我国部分地区蛋鸡群中存在肠炎沙门氏菌的感染,但是鸡群感染率不高。     

Regional distribution of HSP 70 proteins after myocardial infarction

Hypoxia and altered hemodynamic status, both components of myocardial infarction, have been shown to be potent inducers of the 70 kD family of heat shock proteins (HSP70). We hypothesized that after infarction, the surviving myocardium would synthesize HSP70 proteins in a temporally and regionally distinct pattern. We believed that there would be a lack of an HSP70 response in the infarcted area (I), reflecting the loss of viable cells. We further postulated that tissues bordering infarctions (M) would have a compromised HSP70 response. Conversely, we proposed that HSP70 would be induced in septal tissues (S) of the infarcted heart, as a hypertrophic adaptation. A rat model of myocardial infarction was used to examine the changes in relative concentration and distribution of three major HSP70 family proteins; cytoplasmic HSP72, mitochondrial HSP75, and endoplasmic reticular GRP78 (glucose regulated protein) during 21 days of recovery. While all three HSP70 family proteins investigated were detected in all hearts from all groups at all time periods, experimental treatment (infarction) induced changes in relative protein concentrations that varied with time and sample site location. Relative concentrations of HSP72 and GRP78 were unchanged in the 24 h following infarction while relative HSP75 concentrations were halved in M tissues during the same time period. Between days 5 and 7, several changes were noted. M samples displayed nearly twice the relative concentrations of HSP75 and GRP78 after infarction, but showed no change in HSP72. S tissues showed two-fold or larger increases in all three HSP70 family proteins. I samples showed unanticipated increases in HSP75 and GRP78 during this time period. After 14 to 21 days of recovery, HSP70 family protein concentration levels in M, S, and I tissues from infarcted hearts had returned to levels similar to those seen in control animals. We conclude that the myocardium is unable to, or does not, mount an immediate HSP70 response after infarction but does recover such activity by 5–7 days after infarction.This study was supported in part by a grant from the American Heart Association-Kansas Affiliate (KS-94-GS-30)     

益脑颗粒对亚急性衰老大鼠慢性脑缺血模型脑组织中HSP70表达的影响

目的 观察益脑颗粒对慢性脑缺血大鼠脑组织中HSP70表达的影响,探讨益脑颗粒治疗慢性脑缺血的作用机制.方法 采用D-gal注射诱导亚急性衰老大鼠模型,永久性双侧大鼠颈总动脉结扎术(2VO)制备亚急性衰老大鼠慢性脑缺血模型.给予高、低剂量益脑颗粒灌胃,并以尼莫地平为对照,免疫组化法观察大鼠脑组织中HSP70表达.结果 模型组脑组织中HSP70表达无明显改变(P>0.05),益脑颗粒高、低剂量组表达升高(P<0.01),尼莫地平组无明显改变(P>0.05).结论 益脑颗粒能通过增加缺血脑组织中HSP70的表达,改善慢性脑缺血.     

抑制HSP70—2基因表达诱导肝癌细胞G2／M期阻滞的机制研究

目的探讨HSP70—2在肝癌组织中的表达，以及抑制HSP70—2表达对肝癌细胞生长和周期影响的详细作用机制。方法免疫组织化学检测HSP70—2在45例肝癌组织和癌旁组织中的表达，Westernblot检测HSP70—2在5种肝癌细胞株中的表达。设计合成针对HSP70—2基因的特异性shRNA，构建真核表达载体HSP70-2shRNA1和HSP70—2shRNA2。转染肝癌细胞后，MTT法检测细胞增殖，流式细胞仪检测细胞周期，Westernblot检测HSP70—2表达及细胞周期相关蛋白p-Cdc2（Tyr15）、Cdc25C、Cdc2和cyclinB1的表达变化。结果免疫组织化学检测显示，45例肝癌组织中有33例（73．3％）HSP70—2蛋白表达阳性，45例癌旁组织中仅4例（8．9％）表达阳性，表达差异有显著性（P〈0．05）。HSP70—2蛋白在HepG2、Bel-402、Huh-7、Hep3B、SMMC-7721肝癌细胞中的表达明显高于其在L02细胞中的表达。HSP70—2shRNA1和shRNA2均能有效抑制肝癌细胞HepG2和Bel-7402中HSP70-2的表达。与controlshRNA组相比，转染HSP70-2shRNA1和shRNA2组HepG2和Bel-7402细胞生长增殖速度均明显减慢，细胞周期表现为处于G1／M期的细胞比例显著增加。Westernblot检测发现，转染HSP70-2shRNA1和shRNA2后肝癌细胞中磷酸化的p-Cdc2（Tyr15）表达增加，Cdc25C、Cdc2和cyclinB1表达显著减少。结论HSPT0-2在肝癌中过表达，抑制HSP70．2表达可以显著抑制肝癌细胞的增殖，诱导细胞周期阻滞，其诱导G，／M期阻滞的机制是通过影响Cdc25C-Cdc2／cyclinB1通路来实现的。     

肺癌患者血浆热休克蛋白70及其抗体滴度分析

目的探讨热休克蛋白70(HSP70)、热休克蛋白70抗体(HSP70-Ab)与肺癌的关系。方法用WesternBlot和Western Blot-ELISA法分别检测肺癌组(n=31)和对照组(n=30)血浆中HSP70和HSP70-Ab的水平。结果肺癌组HSP70平均光密度值(5710.87±639.65)显著高于对照组(4544.76±1178.92),P<0.01;肺癌组HSP70-Ab(1∶80)滴度组阳性检出率(54.84%)高于对照组(26.47%),差异有显著性意义,P<0.05;同对照组相比,肺癌组HSP70-Ab(1∶80)阳性者HSP70光密度值有增高趋势(t=3.1152,P<0.01)。结论HSP70、HSP70-Ab可能参与了肺癌的发病过程;HSP70,HSP70-Ab(1∶80)有望作为肺癌诊断指标之一。     

Decreased serum heat shock protein 60 levels in newly diagnosed immune thrombocytopenia patients

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by peripheral thrombocyte destruction. In some autoimmune disorders, heat-shock proteins (HSP) are suggested to be an important antigenic factor. In this study, we demonstrated the serum free levels of HSP60, HSP70, anti-HSP60, and anti-HSP70 in ITP patients and healthy controls. Twenty-eight newly diagnosed ITP patients, 35 ITP patients in chronic phase, and 25 healthy controls were enrolled to this study. Serum levels of HSP60, HSP70, anti-HSP60, and anti-HSP70 were determined by the ELISA method. Serum HSP60 levels of newly diagnosed ITP patients were significantly decreased when compared with both chronic phase ITP patients and healthy controls. HSP60 levels of ITP patients (both newly diagnosed and chronic phase) with thrombocyte counts more than 30?×?10⁹/L were significantly increased compared with ITP patients with thrombocyte counts less than 30?×?10⁹/L and there was a positive correlation between thrombocyte counts and serum free HSP60 levels in ITP patients. This is the first study demonstrating the extracellular HSP levels in adult ITP patients. HSPs are shown to have a place in the pathogenesis of many autoimmune disorders. Low level of HSP60 may lead to lack of anti-inflammatory response due to less Treg activation, hence, could be a counterpart in the pathogenesis of ITP. Further studies are needed to understand the role of HSPs in the pathogenesis of ITP and whether they can be used for diagnosis, prognosis, and treatment of ITP.     

Tanespimycin monotherapy in relapsed multiple myeloma: results of a phase 1 dose‐escalation study

Tanespimycin, a heat shock protein 90 (HSP90) inhibitor, induces apoptosis in drug‐sensitive and ‐resistant MM cell lines and in tumour cells from patients with relapsed MM. In this phase 1 dose‐escalation study, the safety, plasma pharmacokinetics, and biological/antitumour activity of tanespimycin were evaluated in heavily pretreated patients with relapsed/refractory MM. Tanespimycin (150–525 mg/m²) was given on days 1, 4, 8, and 11 of each 3‐week cycle for up to 8 cycles. Non‐haematological AEs included diarrhoea (59%), back pain (35%), fatigue (38%), and nausea (35%); haematological AEs included anaemia (24%) and thrombocytopenia (21%). One patient (3%) achieved minimal response (MR), with a progression‐free survival (PFS) of 3 months, a 41% decrease from baseline in urine M protein, and a 33% decrease from baseline in serum M protein. Fifteen patients (52%) achieved SD with a median PFS of 2·1 months; 5/15 had reductions in serum M protein ranging from 7% to 38% and in urine M protein ranging from 6% to 91%. Mean HSP70 levels increased from day 1 h 0 to day 1 h 4 with further increases on day 11 h 0 and day 11 h 4, consistent with a therapeutic treatment effect. Tanespimycin monotherapy was well tolerated and demonstrated activity across all doses tested.