1.Identification of heat shock protein hsp70 family genes from Rana amurensis and its expression profiles upon infection.
Tingting LIU ; Jingjing GUO ; Zhaodong CHEN ; Yufen LIU ; Legang JING ; Peng LIU ; Wenge ZHAO
Chinese Journal of Biotechnology 2023;39(4):1710-1730
Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.
Heat-Shock Proteins/genetics*
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Phylogeny
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Amino Acid Sequence
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HSP70 Heat-Shock Proteins/metabolism*
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Stress, Physiological
2.Advances in research of heat shock factor 1 and its relation with expression of heat shock protein.
Zhen-yu ZHANG ; Jun-jie CHEN ; Rong YU ; Ying CEN
Chinese Journal of Burns 2012;28(1):78-80
This article reviews the advances in the research of the structural characteristics and the activating process of heat shock factor 1 (HSF-1), the factors that influence the expression of HSF-1, and the relationship between HSF-1 and the expression levels of NF-κB and heat shock protein (HSP). The expression of HSF-1 could be regulated in several stages in the course of interconversion between its active and inactive status. Unusual expression of HSF-1 mediated by NF-κB can lead to the quantitative and functional change of HSP. The quantitative and functional variation of HSP caused by regulation of HSF-1 could influence the normal synthesis and the pathological changes induced by excessive synthesis of protective proteins in cells under stress. We expect that further research would help elucidate novel pathways about the genesis and evolution mechanism of keloid, and it may finally help to find a novel strategy in the treatment of keloid.
Animals
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DNA-Binding Proteins
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metabolism
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Heat Shock Transcription Factors
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Heat-Shock Proteins
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metabolism
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Humans
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Transcription Factors
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metabolism
3.Expression of HSP70 in peripheral lymphocytes of the patients with allergic rhinitis.
Lisi, LIU ; Chengfeng, XIAO ; Ming, ZHANG ; Lei, CHENG ; Efen, WANG ; Tangchun, WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(3):310-2
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
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HSP70 Heat-Shock Proteins/*biosynthesis
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HSP70 Heat-Shock Proteins/blood
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HSP70 Heat-Shock Proteins/genetics
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Lymphocytes/*metabolism
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Rhinitis, Allergic, Seasonal/*blood
4.Heat shock response in guinea pigs cochlea with gentamicin ototoxicity.
Yue-Qiu NI ; Hao TANG ; Wen-Shuang FU
Chinese Journal of Applied Physiology 2002;18(2):179-182
AIMTo explore the effects of gentamicin ototoxicity on the expression of heat shock protein 70 in guinea pigs cochlea.
METHODSWe used immunohistochemistry staining and image quantitative analysis system, combined with auditory brainstem response (ABR) measurement to investigate the change on the expression of HSP70 in guinea pigs cochlea of gentamicin ototoxicity.
RESULTSThe levels of HSP70 immunoreactivity in guinea pigs cochlea of experimental animals were high including Corti's organ, stria vascularis, medial spiral limbus, spiral ganglion cells and the threshold of ABR was in high correlation with the expression of HSP70 ([ r] > 0.8, P < 0.01).
CONCLUSIONGentamicin can induce expression of HSP 70 in guinea pigs cochlea and protect hearing function.
Animals ; Cochlea ; drug effects ; physiopathology ; Gentamicins ; toxicity ; Guinea Pigs ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; drug effects
5.Expression of HSP70/HSP27 protein in residual lesion after NPC radiotherapy.
Runwen WANG ; Gaofeng ZHOU ; Shengpeng HUANG ; Xueping FENG
Journal of Central South University(Medical Sciences) 2009;34(11):1091-1095
OBJECTIVE:
To analyze HSP70/HSP27 protein expression in the nasopharyngeal carcinoma (NPC) primary tissues and the residual lesion, and to explore its predictive value.
METHODS:
Immunohistochemical experiment was performed to detect the expression of HSP70 and HSP27 in 58 NPC primary tissues: 28 were in the experimental group with the local residual lesion and 30 in the control group without recurring and metastasis within 5 years by conventional radiotherapy.
RESULTS:
The positive expression of HSP70 and HSP27 in the experimental group was 92.9%(26/28) and 53.6%(15/28), while that in the control group was 53.3%(16/30) and 53.3%(16/30),respectively. There was significant difference in the 2 groups. The common positive expression of HSP70 and HSP27 between the 2 groups had significant difference, 50.0% (14/28) in the experimental group and 16.7% (5/30) in the control group, respectively. There was no significant difference in the negative ratio of HSP70 and HSP27 common expression between the 2 groups, 3.6% (1/28) in the experimental group and 10.0% (3/30) in the control group, respectively.
CONCLUSION
HSP70 may have an important role in the radioresistance of NPC, and may predict the residual lesion after radiotherapy.
Adult
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Aged
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Carcinoma, Squamous Cell
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metabolism
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radiotherapy
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Female
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HSP27 Heat-Shock Proteins
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metabolism
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HSP70 Heat-Shock Proteins
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metabolism
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Humans
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Male
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Middle Aged
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Nasopharyngeal Neoplasms
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metabolism
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radiotherapy
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Young Adult
6.Preliminary study on glucose regulated protein 78 kD and heat shock protein 20 differential expression between left-sided colon carcinoma and right-sided colon carcinoma.
Hai-ping PEI ; Xiao-long LI ; Hong ZHU ; Liang ZENG ; Lin-sheng HUANG
Chinese Journal of Gastrointestinal Surgery 2013;16(1):75-79
OBJECTIVETo analyze the differential protein expression of left-sided colon cancer and right-sided colon cancer.
METHODSTissue samples of left-sided colon cancer (n=7) and right-sided colon cancer (n=7) were collected. Tissue protein was abstracted and two-dimensional gel electrophoresis (2-DE) was used to examine the gel images. Peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching with bioinformatics. Immunohistochemical SP method was used for the detection of glucose regulated protein 78 kD (GRP78) and heat shock protein 20 (HSP20) in left-sided colon cancer (n=50) and right-sided colon cancer (n=50) tissues.
RESULTSSixteen differentiating protein spots were identified. Compared with right-sided colon cancer, 10 proteins including GRP78 up-regulated and 6 proteins including HSP20 down-regulated in left-sided colon cancer. Immunohistochemical detection showed that in left and right sided colon cancer, the positive expression rate of GRP78 was 78% (39/50) and 56% (28/50) and the positive expression rate of HSP20 was 34% (17/50) and 72% (36/50), respectively, and the differences were statistically significant (both P<0.05). The positive rate of GRP78 was associated with tumor differentiation, infiltration layer, TNM staging, lymph node metastasis, and liver metastasis, while the positive rate of HSP20 was associated with tumor gross morphology, TNM staging, and lymph node metastasis (P<0.05).
CONCLUSIONSThere are differentially expressed proteins between left-sided colon cancer and right-sided colon cancer, especially for GRP78 and HSP20, which may be the cause leading to the biological differences between left-sided and right-sided colon cancer.
Adult ; Aged ; Colonic Neoplasms ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; HSP20 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Middle Aged
7.Differential expression of ODF1 in human ejaculated spermatozoa and its clinical significance.
Jing CHEN ; Yong WANG ; Xiang XU ; Zhou YU ; Yao-ting GUI ; Zhi-ming CAI
National Journal of Andrology 2009;15(10):891-894
OBJECTIVETo compare the expressions of ODF1 (outer dense fiber of the sperm tail 1) in ejaculated spermatozoa from normozoospermic and asthenozoospermic men with low sperm motility.
METHODSSemen analyses were performed on the semen samples obtained from normozoospermic (n=20) and asthenozoospermic (n=20) volunteers according to the WHO criteria. To rule out the contamination of germ cells and leucocytes, the human ejaculated spermatozoa were purified by a discontinuous Percoll density gradient centrifugation. RT-PCR and Western blot were used to detect the expressions of ODF1 in the spermatozoa from the two groups.
RESULTSRT-PCR showed that the expression of ODF1 mRNA was significantly lower in the spermatozoa from the asthenozoospermic patients than in those from the normozoospermic men (1.35 +/- 0.25 vs. 2.79 +/- 0.28, P < 0.05). Western blot confirmed the results from RT-PCR and revealed an obviously decreased expression of ODF1 in the spermatozoa of the asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.44 +/- 0.26 vs. 3.64 +/- 0.34, P < 0.05).
CONCLUSIONThe expression of ODF1 was significantly decreased in the ejaculated spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.
Asthenozoospermia ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism
8.Expression of Hsp70 and Caspase-3 in rabbits after severe traumatic brain injury.
Jing ZHANG ; Dai-qin TAO ; Hui ZHAO ; Zhi-yong YIN
Chinese Journal of Traumatology 2012;15(6):338-341
OBJECTIVETo investigate the expression of Caspase-3 and Hsp70 in rabbits after severe traumatic brain injury (TBI) and to explore the feasibility of its application in estimation of injury time in forensic medicine.
METHODSA rabbit model of heavy TBI was developed by high velocity impact on the parietal bone with an iron stick. Totally 8 healthy adult New Zealand white rabbits were randomly divided into control group (n equal to 2) and injury group (n equal to 6). Four hours after injury, tissue specimens from the parietal lobe, temporal lobe, occipital lobe, cerebellum and brainstem were harvested to detect the expression of Hsp70 and Caspase-3 by immunohistochemistry. Besides, the gray values of cells positive for Hsp70 and Caspase-3 were analyzed with an image analyzer.
RESULTSImmunohistochemistry staining demonstrated a low level of Caspase-3 and Hsp70 expression in normal control group. While in injury group, both the Caspase-3 and Hsp70 expression was significantly elevated (P less than 0.05). Positive cells gathered around the lesion focus. Occipital lobe and cerebellum had fewer positive cells while temporal and brainstem had the fewest.
CONCLUSIONThe expression of Caspase-3 and Hsp70 at an early stage following severe TBI is characteristic and can be applied to estimate the time of injury.
Animals ; Brain Injuries ; metabolism ; Caspase 3 ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Immunohistochemistry ; Rabbits ; Random Allocation
9.Heat shock protein in male infertility: advances in studies.
National Journal of Andrology 2013;19(5):464-467
Heat shock protein (HSP) is a group of evolutionarily highly conserved cell chaperone proteins involved in the processes of molecular chaperone, cytoprotection, anti-apoptosis and immunoregulation. Recent studies found that HSP is also involved in spermatogenesis, sperm capacitation and fertilization, which play a significant role in male reproduction. Therefore, further studies on the action mechanisms of HSP in male infertility may offer a new insight into the management of the problem.
Heat-Shock Proteins
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metabolism
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Humans
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Infertility, Male
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metabolism
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Male
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Spermatozoa
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metabolism