The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49 +/- 15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89 +/- 10.13 IOD, P < 0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR.
Gene Expression Regulation ; HSP70 Heat-Shock Proteins/*biosynthesis ; HSP70 Heat-Shock Proteins/blood ; HSP70 Heat-Shock Proteins/genetics ; Lymphocytes/*metabolism ; Rhinitis, Allergic, Seasonal/*blood

The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24 +/- 0.11 and 0. 42 +/- 0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P < 0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9 +/- 9.9 vs 44.8 +/- 15.3, P < 0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P < 0.01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.
Bronchi/metabolism ; Bronchi/pathology ; Cells, Cultured ; HSP70 Heat-Shock Proteins/*biosynthesis ; HSP70 Heat-Shock Proteins/genetics ; Muscle, Smooth/cytology ; Pulmonary Disease, Chronic Obstructive/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Smoking

OBJECTIVETo study the expression and the possible role of JWA protein in oxidative stress-induced damage of MCF-7 cells, especially the relationship between JWA and heat shock proteins (HSPs).

METHODSMCF-7 cells were exposed to different concentration of H(2)O(2) (0.01,0.10, 1.00 mmol/L) for different time (10, 30, 60 and 180 min) respectively. DNA damage was detected by using DNA gel electrophoresis. The MTT assay was used to analyze the effect of H(2)O(2) on the cytotoxicity and relative cell proliferation ratio of the cells. The expressions of JWA, HSP70, HSP27 and HSF1 were determined by Western-blot.

RESULTSThe inhibitory effect on MCF-7 cells viability induced by H(2)O(2) was shown a dose-and time-dependent manner and MCF-7 cells proliferation, and was almost completely inhibited by the exposure of H(2)O(2) at 1.00 mmol/L for 180 min. Hydrogen peroxide treatment of MCF-7 cells caused oxidative stress which up-regulated the expressions of JWA, HSP70 and heat shock factor 1 (HSF1) in a dose-dependent manner, and the expression pattern of JWA was very similar to those of HSP70 and HSF1 but not to HSP27.

CONCLUSIONJWA might enhance intracellular defenses against H(2)O(2)-induced oxidative damage in human breast carcinoma cells. JWA is determined functioning as an effective environmental responsive protein and as a parallel molecule of HSP70 actively participates in the signal pathways of oxidative damage which might be regulated by HSF1.

Cell Line, Tumor ; DNA Damage ; drug effects ; DNA-Binding Proteins ; biosynthesis ; Dose-Response Relationship, Drug ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; Heat Shock Transcription Factors ; Heat-Shock Proteins ; biosynthesis ; Humans ; Hydrogen Peroxide ; adverse effects ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; biosynthesis ; Oxidative Stress ; drug effects ; Transcription Factors ; biosynthesis ; Up-Regulation

The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49 +/- 15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89 +/- 10.13 IOD, P < 0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR.
Adult ; Female ; Gene Expression Regulation ; HSP70 Heat-Shock Proteins ; biosynthesis ; blood ; genetics ; Humans ; Lymphocytes ; metabolism ; Male ; Rhinitis, Allergic, Seasonal ; blood

OBJECTIVETo establish a RT-PCR system for detecting mRNA expression level of 4 hsp genes in human cells.

METHODSRT-PCR system was established with gene cloning, gene recombination, in vitro transcription and RT-PCR techniques; The detection of expression level of 4 hsp genes in SW13 cell was carried out with this system.

RESULTSIn SW13 cell, hsp70 and hsp90 alpha were typical heat shock induced genes, while hsp60 and hsp90 beta were efficiently expressed and further induced by heat-shock to various extent.

CONCLUSIONSIn our hands novel RT-PCR system can be used to detect mRNA expression level of 4 human hsp genes.

Chaperonin 60 ; biosynthesis ; genetics ; HSP40 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Leukemia, T-Cell ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured

Adult ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; blood ; HSP90 Heat-Shock Proteins ; biosynthesis ; blood ; Heat-Shock Proteins ; biosynthesis ; blood ; Hepatitis B Core Antigens ; blood ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Liver ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; blood

OBJECTIVETo study the role of heat shock protein (HSP) 60 and HSP70 in the pathogenesis of oral lichen planus (OLP).

METHODSThe immunohistochemical SP methods were used to detected the expression of HSP60 and HSP70 in 62 cases of oral lichen planus, 10 cases of normal oral mucosa, 21 cases of chronic discoid lupus erythematosus (DLE), 10 cases of benign lymphoadenosis of mucosa, 46 cases of leukoplakia. RT-PCR was used to evaluate the expression of HSP60 mRNA and HSP70 mRNA in 12 cases of OLP and 5 cases of normal oral mucosa.

RESULTSHSP60 was expressed at much higher levels in OLP than other groups. The expression of HSP70 decreased in epithelial cells in erosive lichen planus group. HSP60 mRNA and HSP70 mRNA were expressed at high level in OLP.

CONCLUSIONHSP60 and HSP70 play an important role in pathogenesis of OLP.

Chaperonin 60 ; biosynthesis ; genetics ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Lichen Planus, Oral ; metabolism ; pathology ; Mouth Mucosa ; metabolism ; RNA, Messenger ; biosynthesis

To explore the anti-apoptotic role of electroacupuncture (EA) and its molecular mechanisms after cerebral ischemia/reperfusion (IR) of rats, by using animal model of middle cerebral artery occlusion (MCAO), the changes of the cleavage of PARP were observed by Western blot and the mRNA of heat shock protein (Hsp) 70 and Hsp90 beta detected by competitive RT-PCR after cerebral IR and EA treatment. The results were as follows: (1) The cleavage of PARP was increased in ischemic hippocampus, and EA treatment could attenuate the level of the cleavage remarkably; (2) The mRNA expression of Hsp70 was increased in the ischemic cortex and hippocampus and was further increased after EA treatment; (3) The mRNA expression of Hsp90 beta was decreased in ischemic cortex and hippocampus and the decrease was relatively slight after EA treatment. The above results demonstrated EA treatment could protect neurons from apoptosis after cerebral IR. One of the molecular mechanisms was the promotion of the inducible expression of Hsp70 and the improvement of the inhibition of the expression of Hsp90.
Animals ; Apoptosis ; Brain ; metabolism ; Brain Ischemia ; genetics ; metabolism ; Electroacupuncture ; Female ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; genetics ; metabolism

OBJECTIVETo show the changes of heat shock protein 70(HSP70) expression at cellular level in different heat exposure stages and the significance of HSP70 expression in heat exposure organism.

METHODThe heat exposure model was established in rats with artificial hot climatic chamber[34 +/- 1) degree C, RH 60%]. SD rats were randomly divided into control groups(C) and heat exposure groups(A), and into subgroups including 2, 7, 14, 28 d stages from each one of the groups. The immuno-histochemistry was used to detect HSP70 expression in rat liver and cardiac muscle, and photography analytic software was used to analyze HSP70 expression in liver and cardiac cells.

RESULTSThe expression intensity of HSP70 in heat exposure groups(gray values of liver were 137.0 +/- 5.1, 137.0 +/- 5.2, 137.8 +/- 7.1, 139.2 +/- 5.2 respectively; of cardiac muscle 156.1 +/- 4.4, 155.1 +/- 6.2, 155.4 +/- 4.5, 156.2 +/- 5.1 respectively) was stronger than that in control groups(P < 0.01 or P < 0.05) during all stages of the heat exposure; There was no significant difference in expression intensity of HSP70 among various stages of heat exposure; after 2 d of heat exposure, HSP70 expression in cell nuclei of the liver and cardiac muscle cells was stronger than that in cytoplasm in heat exposure group; HSP70 expression in Kupffer's cells of liver was also stronger than that of control(P < 0.05), but not on 7, 14 and 28 d; the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST), creatine kinase(CK) and hydroxybutyrate dehydrogenase(HBD) showed an increase on 2 d and 28 d of heat exposure.

CONCLUSIONThe vital organs would be damaged on 2 d of heat exposure. High expression of HSP70 at this stage may be a marker of cell damage; Increased HSP70 expression on 7-14 d of heat exposure may play an important role in adaptation to heat, while long term(28 d) heat exposure, the protection of HSP70 from tissue damage may not be enough.

Adaptation, Physiological ; Animals ; HSP70 Heat-Shock Proteins ; biosynthesis ; Heat Stress Disorders ; metabolism ; Liver ; metabolism ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

The study was aimed to explore the role of gene JWA, a novel retinoic acid responsible and cytoskeleton associate gene, in regulating committed differentiation of HL-60 cell and the molecular mechanism in the course of differentiation and apoptosis of leukemic cells. By using FCM, the changes of CD13, CD14, CD15, CD11b and cell cycles were detected in HL-60 cells treated with ATRA (10(-6) mol/L), Ara-C (10 ng/ml) and TPA (10(-8) mol/L) respectively. The samples were determined by semi-quantitative reverse transcript-polymerase chain reaction (RT-PCR) and Western blot for the expression of JWA, Bcl-2, HSP27 and HSP70 at day 0, 2, 4, 6, 8. The results showed that HL-60 cells committedly differentiated into granulocyte-, monocyte-, macrophage-like cells. As a result, JWA was up-regulated in a time-dependent manner, while Bcl-2 was down- regulated at the same time. In ATRA and TPA group, the change of HSP70 had positive correlation with JWA, and negative correlation with Bcl-2. The expression of HSP27 was not detected. Contrast to the cells from APL patient, the expression of JWA need not be activated by ATRA in advance. In this study, we also exposed HL-60 cells in higher dose of Ara-C (20 ng/ml), and JWA expression underwent opposite trend comparing with in lower dose of Ara-C (10 ng/ml). It is concluded that JWA may play double important roles in regulating ATRA and TPA-induced differentiation and apoptosis in leukemic cells. The JWA expression had a negative correlation between induction and cytotoxic response. The difference of JWA expressions between HL-60 cell and ANLL patient cells would be involved in different leukemia pathogenesis.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cytarabine ; pharmacology ; HL-60 Cells ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tetradecanoylphorbol Acetate ; pharmacology ; Time Factors ; Tretinoin ; pharmacology