寻常型银屑病患者皮损组织中热休克蛋白的差异表达

目的比较寻常型银屑病患者皮损组织与健康人皮肤组织中热休克蛋白的表达情况。方法收集银屑病患者的皮损组织及健康人的皮肤组织,提取蛋白,进行二维凝胶电泳并分析比较。选择在银屑病皮损组织中明显差异表达的蛋白点,进行质谱和生物信息学分析。结果对选择的20个差异表达的蛋白质点进行质谱和生物信息学分析,鉴定了包括热休克蛋白(PHB,HSPβ1,CH60)在内的8个蛋白点的质谱。结论在银屑病患者皮损组织与正常组织中,差异表达的热休克蛋白可能是病理的标志物。     

Effects of RRR-alpha-tocopherol on leukocyte expression of HSP72 in response to exhaustive treadmill exercise

Previous research revealed an increased expression of HSP72 in leukocytes after vigorous endurance exercise. We questioned whether more intensive but shorter exercise also induces leukocyte HSP72 synthesis. To delineate the role of reactive oxygen species (ROS) in exercise-related HSP72 induction, we additionally examined the effect of RRR-alpha-tocopherol (alpha-toc) on HSP72 expression using a double-blind placebo (P) controlled cross-over design. After supplementation with alpha-toc (500 I.U. daily) or P for 8 days, 9 male subjects performed a combined exhaustive treadmill protocol (total duration 29.4 +/- 2.0 min). HSP72 was assessed on mRNA (RT-PCR) and protein levels (flow cytometry). HSP72 mRNA rose 3 h after exercise only in the P group, but individual differences (alpha-toc - P) did not reveal significant treatment effects. A moderate but significant rise of HSP72 protein occurred in granulocytes up to 48 h after exercise. Three hours post-exercise, granulocyte HSP72 protein was lower when subjects received alpha-toc, but this effect vanished 24 and 48 h post-exercise. Exhaustive treadmill exercise augments HSP72 mRNA in leukocytes and induced a moderate but prolonged response of granulocyte HSP72 protein. These exercise effects are lower when compared to earlier findings obtained after vigorous endurance exercise. ROS seem to be involved, but do not play the major role in the induction of granulocyte HSP72 synthesis after exhaustive exercise.     

Predicting response to radiotherapy: Evolutions and revolutions

Purpose: To review the many changes which have occurred in the past decades in the field of predicting outcome after radiotherapy from biological characteristics of the tumour or normal tissue. This review will also describe the present state of the art and emerging trends for the future.

Conclusions: From using explanted cells, glass electrodes, exogenous proliferation and hypoxia tracers, and others, there has been a move towards monitoring expression and mutation of genes. Initially this was possible for just one or a few genes, but methods are now available which allow genome-wide monitoring at either the DNA or RNA level. The potential advantage of this evolution is not only to predict but also to understand potential causes of failure, allowing more rational and effective interventions. Comparative genomic hybridisation, mRNA expression profiling, microRNA profiling and promoter methylation profiling have all shown promise in finding signatures correlating with outcome, including after treatment involving radiotherapy. Expected trends for the future are: more signatures relevant to radiotherapy will be discovered; signatures will be refined and reduced to their essentials, such that genome-wide screening will give way to tailored signatures, quantifiable by routine non-array technology; more focus will be on assays predicting which pathway-specific radiosensitising drugs will be effective (exploiting tumour weaknesses); more signatures will be subjected to validation in randomised trials; and proteomics, DNA sequencing and imaging methods will play progressively increasing roles.     

Microwave hyperthermia treatment increases heat shock proteins in human skeletal muscle

Objective

To test the hypothesis that microwave hyperthermia treatment (MHT) increases heat shock proteins (HSPs) in the human vastus lateralis muscle.

Methods

Four untrained healthy male volunteers participated in this study. The lateral side of the thigh of one leg (heated leg) was heated with a microwave generator (2.5 GHz, 150 W) for 20 min. At 1 day after the MHT, a muscle sample was taken from the heated leg. A control sample was taken from the unheated leg on another day of the MHT. For both legs, HSP90, HSP72 and HSP27 levels were compared.

Results

The HSP90, HSP72 and HSP27 levels in heated legs were significantly higher than those in control legs (p<0.05).

Conclusions

Application of MHT can increase the levels of several HSPs in human skeletal muscle.Recent papers have proposed that heat shock proteins (HSPs) can prevent contraction‐mediated muscle damage,^1,2 which reduces muscular strength and engenders muscle soreness.³ Therefore, it is expected that a prior increase in HSPs in the skeletal muscle reduces muscle damage and produces better muscle performance.Although HSPs can be increased by many types of stresses, heat stress is often used in the case of skeletal muscle. Indeed, various HSPs are increased by elevating muscle temperature to 40–41°C in rat skeletal muscle.^4,5 However, it has not been investigated whether elevation in muscle temperature increases HSPs in human skeletal muscles. Moreover, the means to increase HSPs have not been established in human skeletal muscle tissue. For these reasons, this study was undertaken to test the hypothesis that microwave hyperthermia treatment (MHT), which is a popular modality of thermotherapy, can increase HSP levels in human skeletal muscle.     

Pre-exercise alkalosis attenuates the heat shock protein 72 response to a single-bout of anaerobic exercise

The heat shock protein 72 (HSP72) response following exercise is well documented, however, little is known on whether the expression may be mediated by the ingestion of ergogenic aids prior to performance. The purpose of this research was to investigate the effect of sodium bicarbonate (NaHCO₃) ingestion on monocyte and lymphocyte expressed HSP72 and oxidative stress for 4-h post exercise. Seven active males (22.3 ± 2.9 years, 181.6 ± 4.5 cm, 78.1 ± 8.1 kg) performed a 4-min ‘all-out’ cycle test following a dose of 0.3 g kg⁻¹ body mass of NaHCO₃, or an equimolar placebo dose of sodium chloride. HSP72 was measured by flow cytometry and oxidative stress was determined via plasma thiobarbituric acid substances (TBARS) analysis. The NaHCO₃ ingestion significantly increased blood pH (p < 0.001), bicarbonate (p < 0.001) and base excess (p < 0.001) pre-exercise. Despite this there was no evidence of a significantly improved exercise performance when compared with the placebo trials (p ≥ 0.26) (means ± SD; average power 292 ± 43 W vs. 291 ± 50 W; peak power 770 ± 218 W vs. 775 ± 211 W; work completed 71 ± 10 kJ vs. 68 ± 10 kJ). Monocyte expressed HSP72 was significantly lower under experimental conditions during the 4-h post-exercise (p = 0.013), as was plasma TBARS (p < 0.001). These findings suggest that pre-exercise alkalosis can attenuate the stress response to a single bout of anaerobic exercise.     

Quantification of tumour response to radiotherapy

In 1979, the World Health Organization (WHO) established criteria based on tumour volume change for classifying response to therapy as (i) progressive disease (PD), (ii) partial recovery (PR), and (iii) no change (NC). Typically, the tumour volume is reported from diameter measurements, using the calliper method. Alternatively, the Cavalieri method provides unbiased volume estimates of any structure without assumptions about its shape. In this study, we applied the Cavalieri method in combination with point counting to investigate the changes in tumour volume in four patients with high grade glioma, using 3D MRI. In particular, the volume of tumour within the enhancement boundary, the enhancing abnormality (EA), was estimated from T(1) weighted images, and the volume of the non-enhancing abnormality, (NEA) enhancing abnormality, was estimated from T(2) relaxation time and magnetic transfer ratio tissue characterization maps. We compared changes in tumour volume estimated by the Cavalieri method with those obtained using the calliper method. Absolute tumour volume differed significantly between the two methods. Analysis of relative change in tumour volume, based on the WHO criteria, provided a different classification using the calliper and Cavalieri methods. The benefit of the Cavalieri method over the calliper method in the estimation of tumour volume is justified by the following factors. First, Cavalieri volume estimates are mathematically unbiased. Second, the Cavalieri method is highly efficient under an appropriate sampling density (i.e. EA volume estimates can be obtained with a coefficient of error no higher than 5% in 2-3 min). Third, the source of variation of the volume estimates due to disagreements between observers, and within observer, is much greater in the positioning of the calliper diameters than in the identification of the tumour boundaries when applying the Cavalieri method. Additionally, the error prediction formula, available to estimate the coefficient of error of Cavalieri volume estimates from the data, allows us to establish more precise classification criteria against which to identify potentially clinical significant changes in tumour volume.     

影像学生物标志物在肿瘤疗效评价中的应用

肿瘤治疗的疗效评价对肿瘤的药物研发和治疗方案的制定意义重大,以往单纯从形态学变化评价肿瘤治疗效果存在局限性。新的影像学技术和方法提供的影像学生物标志物能够从功能、分子水平描述肿瘤治疗后变化,反映肿瘤治疗后在代谢、氧合状态、新生血管形成、增殖及凋亡等方面发生的改变,为肿瘤疗效评价提供新的方法和思路。     

Leukocyte heat shock protein expression before and after intensified training

The purpose of the current research was to test the hypothesis that exercise induced leukocyte heat shock protein (HSP) expression is increased during periods of intensified exercise training. Seven male endurance cyclists carried out tests of maximal oxygen consumption and endurance capacity. These standard exercise tests were carried out prior to and following 6 days of prescribed intensified training. Sampled leukocytes were examined for Hsp27 and Hsp70 expression using a Fluorescence Activated Cell Scanner (EPICS XL, Coulter). During a period of overreaching, as signified by a drop in time to fatigue following the intensified training period (p = 0.02), the number of extracellular Hsp27 positive granulocytes increased in response to the VO(2)max test. Acute, intracellular HSP responses were observed in both baseline and overreached conditions. The present study showed that a period of intensified training caused adaptations in the acute heat shock protein exercise response, reflected by a greater increase of cell surface HSP positive leukocytes following heavy training.     

Oxygen stress effects on proliferation rates and heat shock proteins in lymphocytes

INTRODUCTION: High-oxygen conditions produce oxidants and cause some pathognomonic signs and symptoms in human hosts. However, the biological effects at the cellular level are still unclear. To investigate the cellular response against oxygen stress, we measured the proliferative activity and heat shock protein (HSP) expression of human lymphocytes. METHODS: Hydrogen peroxide (H2O2) was used as a source of reactive oxygen species. The time course of the proliferative/apoptotic response was evaluated from DNA histogram analysis. HSP response was measured by Western blot analysis. RESULTS: Suppression of lymphocyte cluster formation by oxygen stress was observed to be dose-dependent. Induction of apoptotic cell death and retardation of entering the S-phase were also affected according to the extent of oxygen stress. Oxygen stress equivalent to 500 micromol x L(-1) H2O2 or more showed a severe impairment of the cell cycle and that equivalent to 50-100 micromol x L(-1) H2O2 revealed an intermediate effect. Although an increased expression of HSP72/73 was observed in each group 6 h after oxygen stress, only 100 micromol x L(-1) H2O2-treated cells remained at a high expression level after 24 h. CONCLUSION: It is likely that when cells encounter oxygen stress, an increased expression of HSP72/73 and subsequent repair of damaged proteins cause the retardation of cell cycle progression and prevent damaged cells from entering the S phase. But if the damage is too strong, the cells may go into apoptotic cell death. From our results of the HSP72/73 experiment, the threshold that determines the fate of the lymphocytes may be around 100 micromol x L(-1) H2O2.     

Exercise, heat shock proteins, and myocardial protection from I-R injury

Heat shock proteins (HSPs) play a critical role in maintaining cellular homeostasis and protecting cells during episodes of acute stress. Specifically, HSPs of the 70 kDa family (i.e., HSP72) are important in preventing ischemia-reperfusion induced apoptosis, necrosis, and oxidative injury in a variety of cell types including the cardiac myocyte. Evidence indicates that HSP72 may contribute to cellular protection against a variety of stresses by preventing protein aggregation, assisting in the refolding of damaged proteins, and chaperoning nascent polypeptides along ribosomes. Endurance exercise is a physiological stress that can be used to elevate myocardial levels of HSP72. It is now clear that endurance exercise training can elevate myocardial HSP72 by 400-500% in young adult animals. Importantly, an exercise-induced elevation in myocardial HSPs is associated with a reduction in ischemia-reperfusion (I-R) injury in the heart. Although it seems likely that exercise-induced elevations in myocardial levels of HSPs play an important role in this protection against an I-R insult, new evidence suggests that other factors may also be involved. This is an important area for future research.     

Breast cancer subtypes: response to radiotherapy and potential radiosensitisation

Radiotherapy (RT) is of critical importance in the locoregional management of early breast cancer. Over 50% of patients receive RT at some time during the treatment of their disease, equating to over 500 000 patients worldwide receiving RT each year. Unfortunately, not all patients derive therapeutic benefit and some breast cancers are resistant to treatment, as evidenced by distant metastatic spread and local recurrence. Prediction of individual responses to RT may allow a stratified approach to this treatment permitting those patients with radioresistant tumours to receive higher doses of RT (total and/or tumour cavity boost doses) and/or radiosensitising agents to optimise treatment. Also, for those patients unlikely to respond at all, it would prevent harmful side effects occurring for no therapeutic gain. More selective targeting would better direct National Health Service resources, ease the burden on heavily used treatment RT machines and reduce the economic cost of cancer treatment. Unfortunately, there are no robust and validated biomarkers for predicting RT outcome. We review the available literature to determine whether classification of breast cancers according to their molecular profile may be used to predict successful response to, or increased morbidity from, RT. Class-specific biomarkers for targeting by radiosensitising agents are also discussed.Breast cancer is the most common cancer in females with just over 1 million new diagnoses and some 400 000 deaths recorded worldwide each year (World Health Organization). As such, this disease represents one of the most serious and costly health issues. Locoregional treatment of breast cancer has evolved over the last two decades not only in the surgical techniques used but also in the use and delivery of radiotherapy (RT). Surgical management has moved towards breast conserving surgery (BCS) and axillary sentinel lymph node biopsy wherever possible. Surgery aims to remove any disease that has been detected in the breast and regional lymph nodes. Surgery does not, however, remove undetected occult deposits of cancer that may remain within the breast, scar, chest wall or remaining lymph nodes. It is these undetected deposits that may lead to locoregional recurrence (LR) and furthermore to distant metastases. RT has therefore become a well-established adjuvant treatment modality to optimise local control following most BCS [1] and following mastectomy in those patients at high risk of recurrence [2]. According to a recently published National Health Service Breast Screening Programme audit of 17 013 screen-detected breast cancers collected from April 2009 to March 2010, 10 425 (61%) females received adjuvant RT. This includes those diagnosed with both invasive (9223, 88.5%) and non-invasive (1202, 11.5%) breast cancers. Of the 8739 patients who underwent BCS for invasive breast cancer, 8201 (94%) received RT. These data did not include females who presented to symptomatic breast clinics and indicate the considerable volume of patients requiring and receiving adjuvant RT each year in the UK alone. It is no surprise that there have been capacity issues leading to wide variations in the timelines of availability of RT from country to country, and even within the same country. RT achieves high cure rates where metastatic spread has not occurred [3–5] and offers improvements in overall survival (OS) and disease-free survival (DFS) in patients presenting with distant metastases at diagnosis [6]. A reduction in LR has tumour-related gains and also quality of life benefits, such as a reduction in the physical morbidity and psychological consequences of LR. However, as with all adjuvant therapies, there are recognised short-term (those which happen within ∼3 months of completing treatment) and long-term side effects.     

Exercise training and experimental diabetes modulate heat shock protein response in brain

In diabetes, defense systems against cellular stress are impaired. Heat shock proteins (HSPs) function primarily as molecular chaperones. Factors that raise tissue HSP levels may slow progression of diabetes and improve diabetic complications that also affect brain tissue. This study tested the effect of an 8-week exercise training on brain HSP response in rats with or without streptozotocin-induced diabetes (SID). In untrained animals, the HSP levels were not different between SID and non-diabetic groups. Endurance training, however, increased HSP72 and HSP90 protein in non-diabetic rats, whereas SID significantly decreased the effect of training on these HSPs. At the mRNA level, HSP60, HSP90 and GRP75 were increased due to training, whereas HSP72 mRNA was only increased in exercise-trained diabetic animals. Training or diabetes had no effect on protein carbonyl content, a marker of oxidative damage. Altogether, our findings suggest that endurance training increases HSP expression in the brain, and that experimental diabetes is associated with an incomplete HSP response at the protein level.     

Peripheral vascular responses to heat stress after hindlimb suspension

PURPOSE: The purpose of this study was to determine whether hindlimb suspension (which simulates the effects of microgravity) results in impaired hemodynamic responses to heat stress or alterations in mesenteric small artery sympathetic nerve innervation. METHODS: Over 28 d, 16 male Sprague-Dawley rats were hindlimb-suspended, and 13 control rats were housed in the same type of cage. After the treatment, mean arterial pressure (MAP), colonic temperature (Tcol), and superior mesenteric and iliac artery resistances (using Doppler flowmetry) were measured during heat stress [exposure to 42 degrees C until the endpoint of 80 mm Hg blood pressure was reached (75 +/- 9 min); endpoint Tcore = 43.6 +/- 0.2] while rats were anesthetized (sodium pentobarbital, 50 mg x kg(-1) BW). RESULTS: Hindlimb-suspended and control rats exhibited similar increases in Tcol, MAP, and superior mesenteric artery resistance, and similar decreases in iliac resistance during heat stress (endpoint was a fall in MAP below 80 mm Hg). Tyrosine hydroxylase immunostaining indicated similar sympathetic nerve innervation in small mesenteric arteries from both groups. CONCLUSION: Hindlimb suspension does not alter the hemodynamic or thermoregulatory responses to heat stress in the anesthetized rat or mesenteric sympathetic nerve innervation, suggesting that this sympathetic pathway is intact.     

急性缺氧暴露对豚鼠耳蜗热休克蛋白90表达的影响

目的:观察热休克蛋白90(heat shock protein 90,HSP 90)在急性缺氧后早期豚鼠耳蜗中的表达变化特点。方法:利用自制缺氧舱使实验动物缺氧,应用免疫组化SABC法结合图象分析技术检测正常和急性缺氧后早期豚鼠耳蜗HSP 90 的表达。结果:HSP 90在正常和缺氧损伤后豚鼠耳蜗各回均有不同程度的染色,主要阳性部位是螺旋神经节、螺旋器、血管纹和螺旋韧带。但缺氧损伤后早期HSP 90在耳蜗各阳性部位的表达均有增强,其中在螺旋神经节和螺旋器的表达增强最明显(P<0．01)。结论:HSP 90在正常和缺氧豚鼠耳蜗组织均有表达,缺氧可明显诱导HSP 90在耳蜗组织的表达增强。HSP 90可能对缺氧损害后耳蜗功能的恢复起作用。     

Effects of acute and adaptive hypoxia on heat shock protein expression in hepatic tissue

This experimental study was designed to analyze the expression of heat shock protein (HSP) in hepatic tissue induced by acute and adaptive hypoxic hypoxia. Rabbits were exposed to 5000 m simulated altitude at 11% O(2) in a chamber. Total antioxidant status (TAS) plasma content showed a significant decrease in the acute and adaptive hypoxia groups compared with the control group. Regarding TAS, there was no statistically significant difference between the acute and adaptive hypoxia groups. Histopathological evidence of liver injury was observed in study groups. Immunohistochemical analysis showed diffuse HSP70 staining in the hepatocytes in acute hypoxia group. Staining was focal and prominent in pericentral hepatocytes in the adaptive hypoxia group. As HSP expression appeared increased, total injury score increased as well. There was an inverse correlation between HSP and TAS, but it did not reach statistical value. Our results confirmed the expression of HSP in hepatic tissue related to defense against cellular injury in a hypoxia model. It is an early response in acute hypoxia and may decrease in adaptive hypoxia. It seems that HSP is induced, rather than protectively, as an early marker of liver injury. HSP70 induction and overexpression seem to be, at least in part, explained by impaired antioxidant defense mechanisms.     

The effect of the hyperbaric environment on heat shock protein 72 expression in vivo

Heat shock protein 72 (HSP72) is expressed in response to stress and has been demonstrated to follow a diurnal expression pattern within monocytes and is sensitive to changes in core temperature. Numerous studies have shown changes in HSP72 expression within cell lines exposed to hyperbaric conditions. No studies have investigated changes in HSP72 expression in vivo. Six males participated in the study and were exposed to hyperbaric air and hyperbaric oxygen a week apart. Monocyte HSP72 was analyzed by flow cytometry at 09:00, 13:00, 17:00, 21:00 with hyperbaric oxygen or hyperbaric air breathing commencing at 15:00 for 78 min at a pressure of 2.8 ATA. HSP72 under normoxia followed the established trend; however, following the hyperbaric air or oxygen exposure a reduction in detectable HSP72 was observed at 17:00 and 21:00. No changes in core temperature were observed between 13:00 and 21:00 for any condition. The data show that HSP72 expression is impaired following hyperbaric air (HA) exposure, when compared with control or hyperbaric oxygen (HO) exposure.