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Objective@#To investigate the dynamic change of paraquant-induced kidney injury in rats and the protective effect of edaravone.@*Methods@#Eighty SD rats were randomly divided into 4 groups: the normal control group, paraquat poisoning group, edaravone treatment group and edaravone control group. The normal control group of 8 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and the same amount of fluid into the abdominal cavity after 30 minutes. The paraquat poisoning group of 24 rats were given 1 ml of paraquat solution (20 mg/kg) through the abdominal cavity to build poisoning models, and the same amount of 0.9% sodium chloride was injected into the abdominal cavity after 30 minutes. The edaravone treatment group of 24 rats were given edaravone (5 mg/kg) through the abdominal cavity after 30 minutes when the poisoning models were set up. The edaravone control group of 24 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and edaravone (5 mg/kg) was injected into the abdominal cavity after 30 minutes. In addition to the normal control group, the other groups processed 1 times a day to mantain 7 d. On 1, 3, 7, 21 d several rats in each group were excuted and the kidney tissue and serum samples were collected, then each pathological changes of the kidney were observed with light microscopy. Serum creatinine, KIM-1, NGAL were measured by ELISA, the expression of HSP70 protein in kidney were observed with immunohistochemical staining.@*Results@#The pathological examination reveald that the damage of kidney tissue in the paraquat group was the most serious on 3 d, and the damage was consistently alleviated in edaravone treatment group at the same time, renal fibrosisn was unseen in each group until 21 d. Compared with normal control group, there was no statistically significant in edaravone control group (P>0.05) . The KIM-1 in blood and kidney in paraquat poisoning group were markedly increased in 1 d (P<0.05) . The NGAL in blood and creatinine were markedly increased in d7 (P<0.05) . The NGAL in kidney increased over time, but had no statistically difference with the control group (P>0.05) .Compared with paraquat poisoning group, the serum creatinine, KIM-1 in blood and kidney, the KIM-1 in kidney had decreased significantly in edaravone treatment group (P<0.05) . The NGAL in kidney has no statistically significant compared with the poisoning group (P>0.05) . HSP70 expression of kidney tissue in edaravone treatment group had significantly increased in d3 compared with the paraquat poisoning group (P<0.05) .@*Conclusion@#Edaravone can prompt a significant rise of HSP70 in kidney tissue, reduce KIM-1 and NGAL levels, and play a protective role in kidney injury of acute paraquat poisoning.
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Objective The present study investigated the expression of heat shock cognate protein 70 (Hsc70) in the synovial tissues and blood samples of patients with rheumatoid arthritis (RA) to determine the pathological role of this protein in the pathogenesis of the disease.Methods The expression of Hsc70 in synovial membranes was quantitatively analyzed by immunohistochemistry,real-time quantitative PCR and western blotting.The samples from osteoarthritis (OA) and ankylosing spondylitis (AS) were used as controls.The levels of Hsc70 in blood of patients with RA were determined using enzyme linked immunosorbent assay (ELISA) with the samples of the healthy subjects as controls.Statistical analysis was conducted with one-way ANOVA,LSD test and Spearmen's correlation.Results Immunohistochemistry showed that Hsc70 had significantly increased expression in synovial tissues of RA than in the samples of OA and AS.Real-time PCR and western blotting confirmed the above findings.ELISA detected significantly elevated level of Hsc70 in blood of patients with RA as compared with samples from the controls (P＜0.01).Conclusion The study suggests that the up-regulation of Hsc70 may be involved in the pathogenic process of RA.
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Melanoma antigen-encoding gene 3 (MAGE-3) is an ideal candidate for a tumor vaccine although its potency need to be increased. Heat shock proteins (HSPs) represents a potential approach for increasing the potency of DNA vaccines. In the present study, a fusion DNA vaccine composed of Mycobacterium tuberculosis HSP70 and MAGE-3 was constructed and used to immunize C57BL/6 mice against B16 or B16-MAGE-3 tumor cells. The results show that the HSP70-MAGE-3 fusion DNA vaccine enhanced the frequency of MAGE-3-specific cytotoxic T-cells as compared to the MAGE-3 DNA vaccine or the HSP70/MAGE-3 cocktail DNA vaccine (P<0.05). In conclusion, the results indicate that the HSP70-MAGE-3 fusion DNA vaccine can strongly activate MAGE-3 specific cellular immunological reactions and thus significantly inhibit the growth of B16-MAGE-3 tumors, improving the survival of tumor-bearing mice, and the HSP70-MAGE-3 fusion DNA vaccine has a significant therapeutic effect on the tumors that express MAGE-3 antigens.
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Objective: To study the effect of thermo-chemotherapy on lung cancer and its possible mechanism. Methods: H446 cells were subjected to different thermo-chemotherapy strategies: 43°C + Paclitaxel (120 μg/L, thermo-chemotherapy group), 43°C+Paclitaxel (120 μg/L)+SP600125 (20 μmol/L, JNK inhibitor) (thermo-chemotherapy+SP600125 group), thermotherapy (43°C) group, and Paclitaxel (120 μg/L) group; untreated cells served as control. MTT assay was used to measure cell proliferation and Western blotting was used to examine the expression of JNK, p-JNK and HSP70 protein. Results: The proliferation rate of cells in the thermo-chemotherapy group was significantly lower than those in the other 3 groups (all P<0.05). The expression of p-JNK was significantly increased in the thermo-chemotherapy group (P<0.05); SF600125 inhibited the expression of p-JNK and the proliferation of cells in the thermo-chemotherapy+ SP600125 group was elevated (P< 0.05). The expression of HSP70 in the thermo-chemotherapy group was lower than that of the thermotherapy group (P<0.05). Conclusion: Thermotherapy can obviously promote the inhibitory effect of Paclitaxel chemotherapy against the growth of lung cancer cell line H446, probably through activating JNK pathway or inhibiting expression of HSP70 protein.
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Objective To investigate the relation between the expression of HSP70 and p57kip2 in C, CT and Enneking surgical classifi-cation by detecting expression of HSP70 and p57kip2. Methods 30 cases of GCT were collected, including surgical operation specimens and 13 cases of osteochondroma were taken as control group. The expression of HSP70 and p57kip2 were detected, and the relation between the ex-pression of HSP70 and p57kip2 and Enneking surgical classification and prognosis of GCT were analyzed. Results With the increase of tumor Enneking surgical grades, the positive rate of HSP70 was increased and the negative rate of p57kip2 was decreased. Expression rates of HSP70 were 22. 2%, 88.9% ,100% in grades of 1～3 and expression rates of p57kip2 were 88. 9%, 88.9%, 33.3% in grades of 1～3. There was a negative correlation between HSPTo and p.57kip2 expression. Conclusions HSP70 and p57kip2 may play a role in the genesis, and develop-ment of giant cell tumor of bone, and it can be helpful for surgical classification and prognesis of GCT.
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Objective To study the feasibility and safety of limb remote ischemic preconditioning (RIPC) in infants and explore the protective effect on myecardium ischemia reperfusion injury for infants undergoing cardiac operation under cardiopulmonary bypass. Methods 60 infants weight less than 7 kilograms with ventricular septal defect were enrolled into the study. 30 of them (RIPC group) were ischemic preconditioned two times (24 hours and 1 hour preoperatively) by three cycles of iscbemia (5 minutes for each) and reperfusion on the left upper arm using a blood pressure cuff. Serum lactate dehydrogenase (LDH), creatine kinase (CK) and its isoenzyme (CK-MB), and tro-ponin I (TnI) ; malondialdehyde (MDA) and superoxide dismutase (SOD) was preoperatively detected. The expression of heat shock pro-tein 70 (HSP 70) in cardiomyocytes was determined by western blot analysis. The surgical outcome including limb movement and sensory function was also recorded. Results No limb disability or sensory disturbance or no other surgical complications was found in all infants. LDH, CK, TnI at the beginning of operation in RIPC group was higher than those in control group. After operation, leakage of heart enzymes were attenuated in RIPC group, and the serum concentration of enzymes were lower than those in the control group. The RIPC group had low coronary sinus venous concentration of MDA but high SOD. The expression of HSP70 was upregulated in cardiomyocytes of RIPC group. Conclusion The limb RIPC can be done easily and safety in infants, and BIPC can reduce the leakage of myocardial enzymes and upregu-late the expression of HSP, which possess protective effect on myocardial IRI.
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Objective To investigate the expression of COX-2 in multiple myeloma(MM)and the relationship between myeloma cells proliferation and apoptosis.To provide a new prognosis factor and therapeutic target.Methods COX-2 from the 22 newly diagnosed MM,14 relapsed MM and PCNA,HSP70 of the newly diagnosed patients were detected by immunohistochemistry method.Results All the newly diagnosed MM exhibited positive COX-2 immunoreactivity.50% had strong COX-2 and 50% showed weak COX-2.Relapsed MM exhibited strong COX-2.COX-2 was related with serum β2 microglobulin,marrow plasma cells,hemoglobin,PCNA,HSP70(P=0.019,0.003,0.048,0.006,0.034).Conclusion COX-2 was overexpressed in MM.Prognosis of patients with strong COX-2 is poorer than those with weak COX-2.COX-2 may promote the proliferation and inhibit the apoptosis of myeloma cells.
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Objective To investigate the effect of ketamine on the expression of HSP 70 in myocardium in severely burned rats for its possible mechanism of myocardial protection. Methods Seventy-two male Wistar rats were randomly divided into 3 groups: normal control group (group C, n=8), burn injury group (group BI, n= 32) and ketamine group (group K,n=32). 30% Wtal body surface area of Ⅲ degree burn model was developed in group BI and group K. Ketamine 20 mg/kg was injected IM in group K 15 min after the burn model was made. Equal volume of normal saline was given instead of ketamine in group BI. Group C received no treatment. The rats were sacrificed at 3, 6, 12 and 24 h after administration in group BI and group K respectively(8 rats at each time point). Myocardial samples were obtained for determination of the expression of HSP 70 by Western blot analysis. The myocardial ultrastructure was observed at 3 h after administration with electron microscope. Results The myocardial damage was milder in group K than in group BI. The expression of HSP 70 was significantly higher at 3, 6, 12 and 24 h after administration in group K and group BI than in group C(P<0.05).The HSP 70 expression was significantly higher at 3 and 6 h after administration in group K than in group BI ( P<0.05). Conclusion Ketamine can reduce the myocardial injury induced by severe burn through up-regulating the expression of HSP 70 in cardiocytes.
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Objective To investigate the expression and the early diagnostic significance of heat shock protein 70 in acute allograft rejection of liver-transplamed rats. Methods The model of rat orthotopic liver transplantation was made by using a modified "two-cuff technique". The rats were randomly divided into 3 groups. For each group, donors and receptors all included 15 rats respectively. The control group: Wistar to Wistar liver transplantation; The untreated group: SD to Wistar liver transplantation, not receiving any immunosuppressant after liver transplantation; The treatment group: SD to Wistar liver transplantation, receiving intramuscular injection of tacrolimus (FKS06, 2mg kg-1. day-1) after operation. Five rats were executed randomly in every group on the post-transplantation day 3, 5 and 7 and the graft samples were obtained for optical microscopic observation. The expression of HSPT0 in grafts was detected by using immunohistochemical method and RT-PCR. The correlation between acute rejection following liver transplantation and the expression of HSP70 in grafted liver was studied. Results There was no acute rejection examined in the control group. The untreated group showed typical allograft rejection and the rejection activity index (RIA) went up gradually after the operation (P<0.01). The treatment group showed no rejection or borderline allograft rejection. The level of HSP70 was increased transiently after operation, then reduced in the control group (P<0.05). The level of HSP70 in the untreated group was higher than in the control groupand gradually increased with the prolongation of time after transplantation (P<0.01). A significant correlation was found between HSP70 and pathological score in the untreated group (P<0.01). The treatment group showed low levels of HSP70 of all the time. Conclusions The expression of HSP70 in grafts is closely related to the occurrence and development of the acute rejection and can be useful for early diagnosis of acute allograft rejection following liver transplantation.
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Objective To study that Geranylgeranylacetone(GA) induce the expression of heat shock protein 70 expression in hippocampus of Alzheimer's model rats and its effect on learning and memory ability in the model rats induced by Aβ1-42 injection into hippocampus.Method 72 health SD rats were randomly divided into GGA group,model group was injected with Aβ1-42 and control group was injected with normal saline into hippocampus.Y maze test was used to detect the learning and memory ability of the rats at the 7th,14th and 21st day after injection into hippocampus separately.HSP70 expression in hippocampus tissues were detected by RT-PCR and western-blot as soon as Y maze test has been finished.Result The learning and memory ability of the rats in model group decreased significantly at the 14th and 21st day after injection than those in control group(P<0.05),but not too much changed in GGA group(P<0.05).HSP70 expres- sions in hippocampus tissues in model group decreased gradually after injection Aβ1-42,but increased in GGA group at the 7th,14th and 21st day after injection.Conclusion GGA can induce the expression of HSP70 in hippocarnpus of Alzheimer's model rats and meliorate the neuron impairment as well as the learning and memory ability of the rats.
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Objective To investigate the dynamic changes of HSP70 mRNA expression in the liver tissue of rats with traumatic shock and the treatment effects of glycine.Methods The expression of HSP70 mRNA in the liver tissue of treatment group,shock group and control group was detected by ELISA.Pathological changes were observed,and serum ALT and AST were measured.Results The expression of HSP70 mRNA in the liver tissue of rats in the shock group and the treatment group reached peak at the 6th and 12th hour after resuscitation respectively.Serum ALT and AST increased and pathological damage aggravated with time prolonging.Compared with control group,the expression of HSPT0 mRNA in treatment group increased significantly,serum ALT and AST decreased significantly and pathologi- cal damage was significantly relieved(all P<0.05).Conclusion Glycine can increase the expression of HSPT0 mRNA and relieve the secondary damage of liver after traumatic shock.
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OBJECTIVE To study the expressions of heat shock protein 70 and human papillomavirus16E7 protein in human laryngeal squamous cell carcinoma, and their relationship in the genesis of human laryngeal squamous cell carcinoma. METHODS The expressions of HSP70 and HPV16E7 protein were detected by the immunohistochemical method in 78 specimens with laryngeal squamous cell carcinoma, 24 specimens with vocal cord polyps and 10 specimens of normal laryngeal tissues. RESULTS In human laryngeal squamous cell carcinoma, vocal cord polyps and normal laryngeal tissues, the positive expression rates of HSP70 were 69.2 % , 8.3 % and 0 % respectively, with those of HPV16E7 protein being 43.6 % 4.2% and 0 % respectively. There was a significant difference of the expression rate of HSP70 or HPV16E7 protein between the laryngeal squamous cell carcinoma and the vocal cord polyps(P
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Objective To explore the mechanisms on the damage in fetus liver,kidney and brain with administration of mifepristone and mesoproseol.Methods 17 specimens were obtained from the women who volunteered to terminate their pregnancy during 10～28 weeks.According to the fetus weight,they were divided into 3 groups.Immunohistochemistery was induced to investigate the expressions of pur-?,HSP70.Results The expressions of pur-? and HSP70 could be observed in both the treatment group and the control group.pur-? and HSP70 are the targets in detecting the damage of DNA.But pur-? was more sensitive than HSP70.The damage on group Ⅱ is the strongest.Conclusion Mifepristone can lead to the damage to lover,kidney and brain in mid-pregnancy.Its damage will be reducing as the growing of fetus.
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The heat shock proteins (HSPs) are ubiquitous molecules induced in cells exposed to various stress conditions, including carcinogenesis. The HSP70 and HSP27 among HSPs are of special relevance in human cancer inhibiting apoptosis. The aim of this study is to investigate the expressions of HSP70 and HSP27 in hepatocellular carcinoma (HCC) in association to tumor cell proliferation and apoptosis. We examined the expressions of HSP70 and HSP27 by immunohistochemical staining in 71 cases of HCC, and then related their expressions to clinicopathologic parameters and expressions of p53, Ki-67 and Apotag. HSP70 and HSP27 were frequently stained in the cytoplasm and nuclei of tumor cells, but not in the non-neoplastic hepatocytes. Immunoreactivities of HSP70 and HSP27 were observed in 56.3% and 61.9% of HCCs, respectively. HSP70 immunoreactivity correlated with high Ki-67 labeling indices (LIs) (p=0.0159), large tumor size (p=0.0129), presence of portal vein invasion (p=0.0231), and high tumor stage (p=0.0392). HSP27 immunoreactivity significantly related with the subgroup of HBV-associated HCCs (p=0.0003), but not with the others. Both HSP70 and HSP27 immunoreactivities showed no relation to Apotag LIs or p53 immunoreactivity. In conclusion, expressions of HSP70 and HSP27 may play an important role in hepatocarcinogenesis, and especially HSP70 showed a close relationship to the pathological parameters associated with tumor progression and high Ki-67 LIs. Our results could be additional evidence that HSP70 expressions can contribute to not only hepatocarcinogenesis but also tumor progression by promoting tumor cell proliferation.
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Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Hepatocelular/metabolismo , Regulação Neoplásica da Expressão Gênica , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/metabolismo , Neoplasias Hepáticas/metabolismo , Proteínas de Neoplasias/metabolismo , Células Tumorais Cultivadas , Biomarcadores Tumorais/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: Nitric Oxide (NO) is an endogenous mediator first characterized as an endothelium-derived relaxing factor. It is now recognized as a key mediator in many physiological process such as vasodilatation, neurotransmission, host defense, and iron metabolism. However, much remains to be determined about the pathophysiological role of NO in the airway. Peroxynitrite, which is synthesized by NO, is the diret cause of cellular toxicity in inflammatory reaction. In this study, we investigated the cellular toxcity of peroxynitrite by the expression of Heat-shock proteins 70 (HSP 70) in normal human nasal epithelium (NHNE) at the inflammatory conditions MATERIALS AND METHODS: 3-Morpholinosydronimone clorhydrate which is a peroxynitrite donor was mixed in the media of cultured NHNE cell. RESULTS: HSP 70 was expressed at the peroxynitrite environment of cultured NHNE cells and HSP 70 mRNA was detected with a time-dependent increasing pattern. CONCLUSION: Peroxynitrite may have a cytotoxic effect, and inhibition of peroxynitrite synthesis may have an important role for controlling the cytotoxic and inflammatory conditions of rhinitis and sinusitis.
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Humanos , Fatores Relaxantes Dependentes do Endotélio , Células Epiteliais , Proteínas de Choque Térmico HSP70 , Ferro , Metabolismo , Mucosa Nasal , Óxido Nítrico , Ácido Peroxinitroso , Fenômenos Fisiológicos , Rinite , RNA Mensageiro , Sinusite , Transmissão Sináptica , Doadores de Tecidos , VasodilataçãoRESUMO
PURPOSE: The events of cell stress and cell death are linked, with the heat shock proteins (Hsps) induced in response to stress appearing to function at key regulatory points in the control of apoptosis. The purpose of this study was to investigate the effect of arisostatins A on the Hsp70 expression and signal mechanism of its transcription. MATERIALS AND METHODS: We used natural arisostatins A produced by Actinomycete, in Caki cells. We measured the growth rate of cell using trypan blue staining, and the induction of the transcriptional levels of Hsp70 with arisostatins, which was quantified by reverse transcript-polymerase chain reaction (RT-PCR) and transiently transfecting cells with a Hsp70. The induction of the transcriptional levels of Hsp70 with arisostatins A was quantified by RT-PCR and transiently transfecting cells with a Hsp70 promoter-luciferase reporter plasmid. RESULTS: Arisostatins A-induced Hsp70 up-regulation was not prevented by the overexpression of peroxiredoxinI (PrxI), PrxII or treatment of superoxide dismutase and catalase. However, the arisostatins A-mediated expression of Hsp70 was reduced significantly in Caki cells treated by the antioxidant, N-acetylcystein. Inhibition of the Janus tyrosine kinase (JAK) activity with AG490 did not inhibit the arisostatins A-induced Hsp70 up-regulation, suggesting that JAK is not associated with the arisostatins A-mediated Hsp70 expression. The mechanism of Hsp70 induction depends on the activation of heat shock factor-1. However, arisostatins A did not effect the change in the expression levels of heat shock factor-1. CONCLUSIONS: These findings suggested that Hsp directly regulates specific stress-responsive signaling pathways, which may antagonize the signaling cascades that result in apoptosis.
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Apoptose , Carcinoma de Células Renais , Catalase , Morte Celular , Linhagem Celular , Proteínas de Choque Térmico , Temperatura Alta , Proteínas de Choque Térmico HSP70 , Plasmídeos , Proteínas Tirosina Quinases , Choque , Superóxido Dismutase , Azul Tripano , Regulação para CimaRESUMO
We evaluated DNA protection effect of heat shock protein (HSP) against cytotoxic effects of exogenous nitric oxide (NO) and reactive oxygen intermediate (ROI). Cultured human corneal fibroblasts were divided into 4 groups. Control (Group I) was not exposed to a sub-lethal heat treatment. Other 3 groups were exposed to 43 degrees C for 1 hr, then incubated at 37 degrees C during different duration (1, 6, 24 hr, Group II, III, IV, respectively). Expression pattern of HSP 70 was analyzed by Western blot. Cell viability was measured by MTT assay and the relationship between HSP 70 expression and DNA damage was examined by terminal deoxyribonucleotidyl transferase mediated dUTP-digoxigenin nick and labeling (TUNEL) stain and single cell gel electrophoresis. Expression pattern of HSP 70 was dependent on recovery times. Cell viability following heat treatment was significantly increased and the TUNEL positive cell number was decreased at 6 hr. In single cell gel electrophoresis, tail moments were increased in a dose-dependent manner by SNAP and X/XO. Following heat treatment, tail moments showed decreased significantly at 6 hr. These results suggest that induction of HSP 70 by sub-lethal heat treatment is closely related with cytoprotective effects against oxidative stresses in human corneal fibroblasts.
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Humanos , Sobrevivência Celular , Células Cultivadas , Córnea/citologia , Dano ao DNA , Relação Dose-Resposta a Droga , Fibroblastos/citologia , Temperatura Alta , Proteínas de Choque Térmico HSP70/genética , Marcação In Situ das Extremidades Cortadas , Óxido Nítrico/metabolismo , Doadores de Óxido Nítrico/farmacologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , S-Nitroso-N-Acetilpenicilamina/farmacologia , Xantina/farmacologia , Xantina Oxidase/farmacologiaRESUMO
Objective: To investigate hyperthermia enhanced expression of heat shock protein70 (HSP70) in breast cancer drug sensitive cell line MCF7 and multi-drug resistant (MDR) cell line MCF7/ADR, so as to increase cytotoxic activity of immunologic effector cells against the target cells, and to provide an experimental basis for cell immunotherapy based on hyperthermia. Methods: The immunological effector cells were induced and expanded by cytokines. Breast cancer cell lines MCF7 and MCF7/ADR were treated at 42 ℃ for an hour. Then after being incubated for additional 4 hours, 24 hours and 48 hours, the cells were digested. Flow cytometry (FCM) was used to detect the expression of HSP70 on the target cells. MTT assay was employed to evaluate cell proliferation and the cytotoxic activity of immunologic effector cells against target cells. Results: The expressions of HSP70 in both the target cells had significant difference. The expression of HSP70 in MCF7/ADR cells was higher than in MCF7 cells before hyperthermia. After hyperthermia the expression of HSP70 increased by 6 folds in MCF7/ADR cells, and 22 folds in MCF7 cells and was higher than in MCF7/ADR cells at hour 4. The proliferation inhibition fraction of hyperthermia against MCF7 was significantly lower than that of MCF7/ADR. The cytotoxic activity of immunologic effector cells against MCF7 cells was lower than against MCF7/ADR cells before hyperthermia. After hyperthermia the cytotoxic activity of immunologic effector cells against MCF7 cells rose by 86.23%, and was higher than against MCF7/ADR cells (rose by 30.32%). Conclusion: The expression of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR were enhanced significantly by hyperthermia. Cytotoxic activity of immunologic effector cells against both the target cells were increased by hyperthermia. The differential expressions of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR affect the cytotoxic activity of immunologic effector cells.
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Objective To investigate the kinetics processes of TLR2 and TLR4 in CD14~+ monocyte of patients during and after cardiac surgery with cardiopulmonary bypass(CPB) and the effects of dexamethasone(DXM) on the regulation of TLR2 and 4 in CD14~+ monocyte. Methods Twenty patients undergoing elective atrial/ventricular septal defect correction were randomized to received 1 mg/kg dexamethasone or placebo before induction of anesthesia. The CD14~+ monocyte surface TLR2 and TLR4 and the intracellular HSP70 were stained and analyzed by flow cytometry, and plasma level of TNF-?, IL-6, IL-10, NO and MDA were measured at following times: before the dexamethasone or placebo were administer(T1), before starting CPB(T2), immediately after aortic declamping(T3), 30min after aortic declamping(T4), 5h after skin closure(T5) and 24h after skin closure(T6). Results Both the HSP70~+ TLR2~+ monocytes and HSP70~+-TLR4~+ monocytes,the plasma concentration of TNF-?, IL-6, NO and IL-10 were upregulated after introduction (P
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To explore the molecular pathological mechanism of severe brain injury, the brain diffuse axon injury (DAI) model and Mamarou free drop model were produced in rats. Sagittal sections of the brain were processed by immunohistochemical ABC method using the mouse serum against NF68 subunit and HSP70. The medulla oblongata was observed under the microscope and electron microscope. Left parietal lobe of the free drop model was examined with HE and HSP70 immunohistochemistry. At 30 min post injury, the axons in medulla oblongata were seen to be crooked, swollen,and deranged. The myelin sheath became slightly separated, and the NFs in axoplasma were abnormal . At 2~24 h post injury,obvious axonal swelling, disconnection and formation of axonal retraction balls were seen. Obvious separation of myelin sheaths, local disconnection, vacuolization,peripheral aggregation of mitochondria and partial dissolution of axoplasma were seen. The NF68 positive axons increased gradually in staining intensity. HSP 70 positive cells of the two groups were detected at 3h after brain injury, reached the peak at 24h, and decreased at 72h. The HSP expression of the two groups were in accord. The research indicated that DAI could lead to a derangement in structure of NFs. Ischemia and anoxia may aggravate the brain injury.