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

Cadmium ; pharmacology ; Cells, Cultured ; Heat-Shock Proteins ; biosynthesis ; Humans ; Metallothionein ; biosynthesis ; Ubiquitin ; biosynthesis

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

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 examine the inhibition of nitric oxide (NO) synthesis during ischemic preconditioning (IP) upon the induction of heat-shock protein 72 (HSP72) and the size-limiting effect of the second window of protection on infarction.

METHODSRabbits were subjected to either 4 cycles of 5-min long coronary artery occlusion separated by 10 min of reperfusion, or a sham operation. During this procedure, we administered 10 mg/kg of N(G)-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase) intravenously 5 min before IP followed by its continuous infusion (1.5 mg/kg/min). Twenty-four hours after IP or the sham operation, the hearts were rapidly excised for assay of HSP72 expression or were subjected to 30 min of coronary artery occlusion followed by 120 min of reperfusion, at which point infarct size (IS) was measured.

RESULTSTwenty-four hours after IP or the sham operation, there was no difference in heart rate or mean arterial pressure between the groups. Immunoblotting revealed an increase in HSP72 protein levels in the IP group, which was blocked by L-NAME. IS in the IP rabbits was reduced compared with controls (29.8 +/- 3.7% vs. 50.8 +/- 4.3%, P < 0.01). IS in the IP rabbits was elevated as a result of L-NAME treatment (46.0 +/- 5.1%). Administration of L-arginine reversed the effects of L-NAME on the induction of HSP72 and IS (33.5 +/- 4.0%). The intravenous administration of S-nitroso-N-acetylpenicillamine (an NO donor, 15 microg/kg/min) over 20 min increased the induction of HSP72 and reduced IS (31.3 +/- 5.7%, P < 0.01 vs. control) 24 h later.

CONCLUSIONThese findings suggest that NO may be involved in the induction of HSP72 and the opening of the second window of protection of IP.

Animals ; HSP72 Heat-Shock Proteins ; Heat-Shock Proteins ; biosynthesis ; Hemodynamics ; drug effects ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Infarction ; pathology ; Nitric Oxide ; physiology ; Penicillamine ; analogs & derivatives ; pharmacology ; Rabbits

OBJECTIVETo examine the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia (BPH) and prostatic cancer (PCa) tissues and to analyse the relationship between their expression and BPH and PCa, especially the relationship with the occurrence, development, prognosis and treatment of PCa.

METHODSAn immunohistochemical staining (SP method) for HSP27 and c-kit was undertaken on 40 BPH and 40 PCa tissues samples.

RESULTSConsistent patterns of cytoplasmic staining for HSP27 were seen in all sections of tissue from BPH. The glandular epithelium stained very strongly positively and the stroma stained positively. The staining for HSP27 in PCa tissues was located in the cytoplasm of glandular epithelia, but the expression of HSP27 in PCa was higher than BPH (P < 0.05). The staining for c-kit in BPH tissues was located in the cytoplasm of smooth muscle cells, and in PCa tissues was located in epithelial cells. The expression of c-kit in PCa tissues was lower than BPH (P < 0.05). The expression level of both HSP27 and c-kit were decreased with the development of grade of PCa (P < 0.05); HSP27 was increased with the development of clinical stage of PCa (P < 0.05 ); c-kit was decreased with the development of clinical stage of PCa (P < 0.05).

CONCLUSIONThe expression level of HSP27 and c-kit was highly correlated with the process of the development from BPH to PCa, and also correlated with tumor grades and stages. The expression of HSP27 and c-kit may be used as an important pathological index and may be helpful for the treatment of PCa.

Aged ; Aged, 80 and over ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins ; biosynthesis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; biosynthesis

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

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

The purposes of this study were to assess the expression patterns of heat shock proteins (Hsps), after eyeball heating or cooling, and to elucidate their relationships with corneal wound healing and intraocular complications after excimer laser treatment. Experimental mice were grouped into three according to local pretreatment type: heating, cooling, and control groups. The preconditioning was to apply saline eyedrops onto the cornea prior to photoablation. Following photoablation, we evaluated corneal wound healing, corneal opacity and lens opacity. Hsp expression patterns were elucidated with Western blot and immunohistochemical staining. The heating and cooling groups recovered more rapidly, and showed less corneal and lens opacity than the control group. In the heating and cooling groups, there were more expressions of Hsps in the cornea and lens than in the control group. These results were confirmed in the Hsp 70.1 knockout mouse model. Our study showed that Hsps were induced by the heating or cooling preconditioning, and appeared to be a major factor in protecting the cornea against serious thermal damage. Induced Hsps also seemed to play an important role in rapid wound healing, and decreased corneal and lens opacity after excimer laser ablation.
Animals ; Blotting, Western ; Cornea/pathology ; Heat ; Heat-Shock Proteins/biosynthesis ; Heat-Shock Proteins 70/genetics ; Immunohistochemistry ; Keratectomy, Photorefractive, Excimer Laser/*methods ; *Lasers ; Lens, Crystalline/pathology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Support, Non-U.S. Gov't ; Temperature ; Time Factors ; Wound Healing