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Expression of endoplasmic reticulum molecular chaperon GRP94 in human lung cancer tissues and its clinical significance

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Chinese Medical Journal 2002年第11期115卷 1615-1619,145页

作者：王琪安利佳陈誉华岳世昌大连理工大学生物工程系大连116012

OBJECTIVE: To investigate the relationship between the expression of glucose regulated protein 94 (GRP94) at the level of mRNA and protein in vivo and in human lung cancer. METHODS: RT-PCR, immunohistochemistry and/or Western blot were used in 54 cases of lung cancer tissues and corresponding normal lung tissues. RESULTS: There was a significant overexpression of GRP94 mRNA and protein in lung cancer tissues as compared with lung normal tissues. In lung cancer tissue, the relative level of GRP94 mRNA as evaluated by RT-PCR was 3.48 +/- 2.06, the level of GRP94 protein as evaluated by immunohistochemistry was + + to + + +, and by Western blot was 256.7 +/- 80.6. In lung normal tissue, the relative level of GRP94 mRNA was 2.01 +/- 1.83, the level of GRP94 protein was + to + + and 108.1 +/- 42.3. The differences in expression of GRP94 between the two tissues were significant (P

关键词： Adult Aged Blotting, Western Endoplasmic Reticulum Female hsp70 heat-shock proteins Humans Immunohistochemistry Lung Lung Neoplasms Male Membrane proteins Middle Aged Neoplasm Staging RNA, Messenger

Effects of Batroxobin on Spatial Learning and Memory Disorder of Rats with Temporal Ischemia and the Expression of hsp32 and hsp70

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Journal of Traditional Chinese Medicine 2000年第4期20卷 297-301页

作者：吴卫平匡培根姜树军张小澍杨炯炯隋南 Albert Chen 匡培梓 Neurotransmitter Research Laboratory Department of NeurologyChinese PLA General HospitalPostgraduate Military Medical SchoolBeijing 100853 Neurotransmitter Research Laboratory Department of NeurologyChinese PLA General HospitalPostgraduate Military Medical School Beijing 100853 Neurotransmitter Research Laboratory Department of NeurologyChinese PLA General Hospital Postgraduate Military Medical SchoolBeijing 100853 PLA 261 Hospital Beijing 100094 Institute of Psychology Chinese Academy of SciencesBeijing 100101

　　The effect of Batroxobin on spatial memory disorder of left temporal ischemic rats and the expression of hsp32 and hsp70 were investigated with Morri`s water maze and immunohistochemistry methods. The results showed that the mean reaction time and distance of temporal ischemic rats in searching a goal were significantly longer than those of the sham-operated rats and at the same time hsp32 and hsp70 expression of left temporal ischemic region in rats was significantly increased as compared with the sham-operated rats. However, the mean reaction time and distance of the Batroxobin-treated rats were shorter and they used normal strategies more often and earlier than those of ischemic rats. The number of hsp32 and hsp70 immune reactive cells of Batroxobin-treated rats was also less than that of the ischemic group. In conclusion, Batroxobin can improve spatial memory disorder of temporal ischemic rats; and the down-regulation of the expression of hsp32 and hsp70 is probably related to the attenuation of ischemic injury.

关键词： Oxygenases Animals Batroxobin Brain Ischemia Down-Regulation hsp70 heat-shock proteins heat-shock proteins Heme Oxygenase (Decyclizing) Learning Disorders Male Maze Learning Memory Disorders Random Allocation Rats Rats, Wistar Snake Venoms Spatial Behavior Temporal Lobe

Purification of heat shock protein 70-associated tumor peptides and their antitumor immunity to hepatoma in mice

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World Journal of Gastroenterology 2004年第3期10卷 361-365页

作者： Dai-XiongChen Yan-RongSu Gen-ZeShao Zhen-ChaoQian KeyLaboratoryofCellEngineeringofGuizhouProvince AffiliatedHospitalofZunyiMedicalCollegeZunyi563003GuizhouProvinceChina DepartmentofPathophysiologyandDepartmentofBiochemistry ShantouUniversityMedicalCollegeShantou51503lGuandonProvinceChina ResearchCenterofCancerBiotherapy InstituteofCancerResearchDalianMedicalUniversityDalian116027LianingProvinceChina

AIM:To purify the heat shock protein (hsp) 70-associated tumor peptides and to observe its non-MHC-I molecule restrictive antitumor ***:By ConA-sepharose affinity chromatography,ADP-agarose affinity chromatography, and DEAE anion exchange chromatography, we were able to purify hsp70-associated peptides from mouse hepatoma (HCaF) cells treated in heat shock at 42℃. Specific active immunization and adoptive cellular immunization assay were adopted to observe the immunoprotective effect elicited by hsp70-associated peptide complexes isolated from ***: The finally purified hsp-associated peptides had a very high purity and specificity found by SDS-PAGE and Western blot. Mice immunized with hsp70-associated peptide complexes purified from HCaF cells were protected from HCaF living cell challenge. This effect was dose *** immunization of immune spleen cells of mice immunized with hsp70-associated peptide complexes could elicit immunity against HCaF challenge, and the tumor-free mice could resist repeated challenges. This effect could be continuously enhanced by repeated challenge with HCaF living cells. The tumor-free mice could tolerate the challenge for as high as 1×10^7 HCaF cells. The mice immunized once with spleen cells pulsed with hsp70-associated peptide complexes in vitro could also result in a certain adoptive immunity against ***:High purity and specificity of hsp70-associated peptides could be achieved from tumor cells by the low-pressure affinity chromatography method used in this study. hsp70-associated peptide complexes derived from the HCaF can elicit non-MHC-I molecule restrictive immunoprotective effect against *** effect can be transferred by adoptive immunization to mice and enhanced by repeated challenge with HCaF live cells.

关键词： Animals Carcinoma, Hepatocellular hsp70 heat-shock proteins purification Immunity Liver Neoplasms, Experimental Mice Mice, Inbred BALB C Peptide Fragments Research Support, Non-U.S. Gov't

Sleep deprivation increase the expression of inducible heat shock protein 70 in rat gastric mucosa

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World Journal of Gastroenterology 2001年第4期7卷 496-499页

作者： Xi-Zhong Shen Marcel W.L. Koo Chi-Hin Cho Department of Gastroenterology,Zhongshan Hospital,Fudan University,136 Yixueyuan Road,Shanghai 200032,ChinaDepartment of Pharmacology.Faculty of Medicine,University of Hong Kong,5 Sassoon Road,Pokfulam,Hong Kong,ChinaSupported by .Dr.Marcel W.L.Koo,Department of Pharmacology,FacuLty of Medicine,the University of Hong Kong,5 Sassoon Road,Hong Kong,China.Wlkoo@hkusua.hku.hk Department of Pharmacology Faculty of Medicine The University of Hong Kong 5 Sassoon Road Hong Kong China.

AIM: To investigate if sleep deprivation is able to increase the expression of inducible heat shock protein 70 in gastric mucosa and its possible role in mucosal defense. METHODS: Rats for sleep disruption were placed inside a computerized rotating drum, gastric mucosa was taken from rats with 1, 3 and 7d sleep deprivation. RT-PCR, immunohistochemistry and Western blotting were used to determine the expression of heat shock protein 70. Ethanol (500mL.L(-1), i.g.) was used to induce gastric mucosa damage. RESULTS: RT-PCR, Western blotting and immunostaining confirmed that the sleep deprivation as a stress resulted in significantly greater expression of inducible heat shock protein 70 in gastric mucosa of rats. After the 500mL.L(-1) ethanol challenge, the ulcer area found in the rats with 7d sleep deprivation (19.15 +/- 4.2)mm(2) was significantly lower (Pheat shock protein 70 in gastric mucosa of rats, the heat shock protein 70 may play an important role in gastric mucosal protection.

关键词： Animals Central Nervous System Depressants Ethanol Gastric Mucosa Gene Expression hsp70 heat-shock proteins Immunohistochemistry Male RNA, Messenger Rats Rats, Sprague-Dawley Research Support, Non-U.S. Gov't Sleep Deprivation Stomach Ulcer control

Zinc is a potent heat shock protein inducer during liver cold preservation in rats

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Chinese Medical Journal 2002年第12期115卷 1777-1779,144-145页

作者：程颖刘永锋梁健中国医科大学第一临床医院器官移植科沈阳110001

OBJECTIVE: A simple liver cold preservation model was established to study the synthesis of heat shock protein 70 (hsp70) induced by zinc (ZnSO(4), i.p.) and its protection during liver cold preservation in rat. METHODS: Male Wistar rats were divided into 5 groups (n = 6). In control group rat received no pretreatment; in Zn-1 group, Zn-2 group, and Zn-3 group rats were pretreated with zinc sulfate at a dose of 5 mg/kg, 10 mg/kg, 15 mg/kg respectively; and in H group rat received heat shock preconditioning (42.5 degrees C x 15 min). Livers were preserved in UW solution for 6, 12 and 24 h, respectively. hsp70 was analyzed by Western blot. Aspartate transaminase (AST) and lactate dehydrogenase (LDH) values of the perfusion solution and the histology of the liver were evaluated. RESULTS: hsp70 expression was markedly elevated after pretreatment with zinc and heat shock. AST and LDH values in the Zn-1, Zn-2 and H groups were significantly lower than those in the control group, respectively (P 0.05), whereas the AST and LDH values in the Zn-3 group were much higher than those in the control group. Histology results showed that liver injury in the Zn-1, Zn-2 and H groups were minimal, while it was severe in the Zn-3 group. CONCLUSIONS: Zn(2+) is a potent and feasible inducer of hsp expression and is able to protect liver from cold preservation injury. The proper inducing dosage of Zn(2+) ranged from 5 mg/kg to 10 mg/kg. The dosage of 15 mg/kg for Zn(2+) as a hsp inducer is not indicated for its severe toxicity to the liver.

关键词： Organ Preservation Animals Aspartate Aminotransferases Dose-Response Relationship, Drug hsp70 heat-shock proteins L-Lactate Dehydrogenase Liver Male Rats Rats, Wistar Zinc

RADIX SALVIAE MILTIORRHIZAE PROTECTS RAT HIPPOCAMPAL NEURON IN CULTURE FROM ANOXIC DAMAGE

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Journal of Traditional Chinese Medicine 1998年第1期18卷 49-54页

作者：刘军匡培根吴卫平张凤英刘洁晓王福庄黄燕华丁爱石 Neurotransmitter Research Laboratory Chinese PLA General Hospital Postgraduate Military Medical School Beijing 100853 China Research Laboratory of Neurobiologe Chinese Military Medical Academy of Sciences Beijing 100850 China

Radix Salviae Miltiorrhizae (RSM), a well-known traditional Chinese medicinal herb, has been used to improve blood circulation and resolve blood stasis. We have previously found that RSM has neuroprotective effect on ischemia and/ or ischemia-reperfusion rats. The purpose of this study was to obtain further information on the mechanism of the RSM-in-duced neuroprotection and to examine the neuroprotective effect on neurons exposed to anoxia. The effect of RSM on anoxic damage in cultured hippocampal neurons of neonatal rat was investigated by using morphological changes and heat shock protein 70kD (hsp70) expression as indicators. RSM given 0.5h before 2h-anoxia followed by 48 hours reoxygenation could significantly increase survival rate of hippocampal neurons and number of hsp70 positive cells. The results suggest that RSM has a direct neuroprotective effects on anoxic damage in hippocampal neurons.

关键词： Animals Animals, Newborn Cell Hypoxia Cells, Cultured Drugs, Chinese Herbal hsp70 heat-shock proteins Hippocampus Neurons Neuroprotective Agents Rats Rats, Wistar Reperfusion Injury

Batroxobin Against Anoxic Damage of Rat Hippocampal Neurons in Culture:Morphological Changes and hsp70 Expression

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Journal of Traditional Chinese Medicine 2001年第3期21卷 215-219页

作者：刘军匡培根吴卫平王福庄丁爱石 Neurotransmitter Research Laboratory Chinese PLA General HospitalPostgraduate Military Medical SchoolBeijing 100853 Research Laboratory of Neurobiology Military Medical AcademyBeijing 100850

Batroxobin,the thrombin-like enzyme,is used for therapeutic defibrination. We have found that batroxobin has good therapeutic effect in ischemic reperfusion rats and clinical practices in vivo. But we have not studied the neuroprotective effect of batroxobin on anoxic hippocampal neurons in vitro. The purpose of this study was to obtain further information on the mechanism of the batroxobin-induced neuroprotection and examine the neuroprotective effect on neurons exposed to anoxia. The effect of batroxobin on anoxic damages in cultured hippocampal neurons of neonatal rats was investigated by using morphological changes and heat shock protein 70Kd (hsp70) immunoreactive expression as indicators. The results indicate that batroxobin, besides its defibrination, may have a direct neuroprotective effect on anoxic damage of hippocampal neurons.

关键词： Animals Animals, Newborn Batroxobin Cell Hypoxia Cell Survival Cells, Cultured hsp70 heat-shock proteins Hippocampus Neurons Neuroprotective Agents Rats Rats, Wistar

Prokaryotic expression and purification of fusion protein hsp70-EGFP and its application in the study of dendritic cells internalization antigen

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Frontiers of Medicine 2008年第1期2卷 82-86页

作者： Ping QU Yanfang SUI Libing LIU Jiahai MA Guangsheng CHEN Jiankang CHEN Fang’e LIU Department of Pathology School of Basic MedicineFourth Military Medical UniversityXi’an 710033China Center of Basic Medicine and Teaching Experiment School of Basic MedicineFourth Military Medical UniversityXi’an 710033China

To study the endocytic activity of dendritic cells(DCs)by obtaining fusion protein hsp70-EGFP as exogenous antigen and loading it with DCs derived from human peripheral *** protein hsp70-EGFP was prokaryotically expressed,isolated and *** were isolated and cultured from human peripheral *** DCs were divided into 3 groups in the endocytic *** were 106 DCs in each *** 1 and 2 were respectively incubated for 30 *** hsp70-EGFP and *** 3 was incubated with hsp70 for 30 min,and then incubated for 30 *** ***,3 groups were placed in an incubator at 37℃for 0.5,1,2 and 24 *** cytometry(FCM)was adopted to detect the amount of DCs with EGFP ***-12 Eli-spot was adopted to detect the amount of DCs which secreted *** were 5 types in the experiment:LPS,inactive LPS,hsp70-EGFP,EGFP and no *** protein hsp70-EGFP was successfully obtained and its molecular weight was *** accounted for 35.32%of the total *** irradiation of an ultraviolet lamp,the protein solution sent out viridescent *** result detected by FCM indicated that after incubation for 0.5 h at 37℃,the positive rate in group 1 was 63%,while the other 2 groups were *** incubation for 1,2 and 24 h at 37℃,the positive rates in the 3 groups were above 80%.The IL-12 Eli-spot examination shows that with hsp70-EGFP being loaded,the amount of DCs secreting IL-12 was 134.09±31.78/10^(5)cells,a little lower than that of DCs with LPS loaded(with the average point of 156.36±15.73).There was no significant difference between the 2 groups(P<0.01).By contrast,both of them were significantly higher than inactive LPS-(33.78±1.40)/10^(5)cells and EGFP-loaded(23.13±4.57)/105 cells DC groups in the amount of DCs secreting IL-12(P<0.01).The results suggest that receptor-mediated phagocytosis plays a main role in the preliminary stage of DCs internalizing *** increasing incubation time,pin

关键词： receptor mediated phagocytosis dendritic cells fusion protein hsp70 heat-shock proteins enhanced green fluorescent protein

Effect of moxibustion on expressions of hsp70 mRNA and protein in gastric cancer-bearing rats

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Journal of Acupuncture and Tuina Science 2019年第6期17卷 395-401页

作者： Tan Jing Lin Ya-ping Yi Shou-xiang Zhao Huan Peng Zhuo-jun Ouyang Li-zhi Peng Yan Hunan University of Chinese Medicine Changsha 410208China

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70(hsp70)in gastric cancer-bearing ***:A total of 40 healthy Sprague-Dawley(SD)rats were adaptively fed for one *** gastric cancer model was prepared by Walker-256 cancer tissue *** 7 d,10 rats were randomly selected to verify the successful modeling,and the remaining 30 rats were divided into a model group,a moxibustion group and an infrared group by the random number table method,with 10 rats in each *** enrollment,the moxibustion group received suspended moxibustion at Zhongwan(CV 12),Guan yuan(CV 4)and bilateral Zusanli(ST 36),(the first group of acupoints)on the 1st day,and suspended moxibustion at bilateral Pishu(BL 20)and Weishu(BL 21),(the second group of acupoints)on the 2nd day,20 min each time,once a *** was alternately performed every other day at the two groups of acupoints for 21 *** the day of en roll me nt,rats in the infrared group were irradiated with the infrared radiatio n at the stomach area on the 1st day,and at the T12-Ti3 interspinous region on the 2nd day,20 min each time,once a day,and the two locations were alternately irradiated every other day for 21 *** the treatment,rats in the model group were intervened by grasping and fixation without *** the end of the treatment,blood was collected from the inner eye orbit,and the hsp70 expression in peripheral blood was determined by enzyme linked immunosorbent assay(ELISA).Rats were sacrificed,the tumor volume and growth inhibition rate were *** position and changes of hsp70 in gastric cancer were observed by streptavidin-perosidase(SP);hsp70 protein expression was determined by ELISA;hsp70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction(R「PCR)***:In comparison of the model group,the volume growth of the gastric cancer in the moxibustion group was signific

关键词： Acupuncture-moxibustion Therapy Moxibustion Therapy Moxa Stick Moxibustion Neoplasms Stomach hsp70 heat-shock proteins Rats