The mRNA Expression Profiles of Five Heat Shock Protein Genes from Frankliniella occidentalis at Different Stages and Their Responses to Temperatures and Insecticides

The westem flower thrips, Frankliniella occidental＆ （Pergande） is a highly invasive pest that is able to exploit many crops across a wide range of environmental conditions. Five full-length cDNAs of heat shock protein （HSP） genes （Fo-HSP90, Fo-HSP70, Fo-HSP60, Fo-HSP40 and Fo-HSP28.9） were cloned from F. occidentalis, and their expression profiles were investigated under conditions of thermal stress and insecticide exposure, and at different stages during development, using real-time quantitative PCR. All five gene sequences showed high similarity to homologs in other species, indicating the conserved fimction of this gene family. HSP60 represents an informative phylogenetic marker at the ordinal taxonomic level within Insecta, but HSP90, which has two homologous copies in Hymenoptera, was not informative. The expression of Fo-HSPs under thermal stress suggests that Fo-HSP90, Fo-HSP70, and Fo-HSP28.9 are inducible by both cold and heat stress, Fo-HSP40 is only heat-inducible, and Fo-HSP60 is thermally insensitive. There were two patterns of cold induction of Fo-HSPs： one is from 0 to 4℃ and the other is around -8℃. All five Fo-HSPs genes were induced by exposure to sublethal concentrations of the insecticide avermectin. The expression of the five Fo-HSPs during different developmental stages suggests that they all play a role in development of F. occidentalis.

Expression of Foreign Gene in Mycobacterium Regulated by Human Mycobacterium Tuberculosis Heat Shock Protein 70 Promoter

The DNA fragments of 150bp length promoter 0f human Mycobacterium(M.) tuberculosis heat shock protein (hsp)7O and 650bp length foreign gene, the Schistosoma japonicum glutathione S-transferase(Sj26GST)gene,were obtained by amplification with polymerase chain reaction. And the 150p DNA sequence upstream initiation codon ATG of the human M. tuberculosis hsp7O promoter that contains the sequence TTGAG and ATCATA which consensus with E. coli promoter's -35 and-10 region respectively, as well as ribosome binding site GGAGG at position-12-8 upstream the ATG were determined by SangerDideoxyribonucleotide-mediated chain-termination method-Then, the human M. tuberculosis hsp70 promoter and Sj26GST cDNA were cloned into E. coli-mycobacteria shuttle plasmid pBCG-2000 to construct E. coli-Mycobacterium expression shuttle plasmid pBCG- Sj26 that can express Sj26GST gene.The M. smegmatis were electroporated and the positivecolonies were selected by kanamycin-The M.smegmatis containing the vector pBCG-Sj26 can be induced by heating and hydrogen peroxide (H2O2) to express GST. The molecular weight of the recombinant GST(rGST) was 26000. The rGST contents that were about 10 percent of the total bacterial protein were analyzed by density scanning after running SDS-PAGE. This study would provide scientific evidences for application of hsp70 promoter in expressing foreign gene in mycobacterium and development of mycobacterium as multiple-valent vectoral vaccine.

Effect of the C.–1388 A>G polymorphism in chicken heat shock transcription factor 3 gene on heat tolerance

Heat stress is one of the main factors that inlfuence poultry production. Heat shock proteins (HSPs) are known to affect heat tolerance. The formation of HSPs is regulated by heat shock transcription factor 3 (HSF3) in chicken. A DNA pool was established for identifying single nucleotide polymorphisms (SNPs) of the chicken HSF3, and 13 SNPs were detected. The bioinformatic analysis showed that 8 SNPs had the capacity to alter the transcription activity of HSF3. The dual luciferase report gene assay showed that there was a signiifcant difference (P<0.01) in the Firelfy luciferase/Renil a luciferase ratio (F/R) of C.–1 703 A>G (S1) and C.–1 388 A>G (S4) sites at the 5′-untranslated region (UTR) of chicken HSF3. The elec-trophoretic mobility shift assay showed that the S4 site was a transcription binding factor. The analysis of the association of the S1 and S4 sites with heat tolerance index revealed that the S4 site was signiifcantly correlated with the CD3+T cel , corticosterone, and T3 levels in Lingshan chickens and with the heterophil/lymphocyte value in White Recessive Rock. These results showed that the S4 site at the 5′ UTR of chicken HSF3 might have an impact on heat tolerance in summer and could be used as a potential marker for the selection of chicken with heat tolerance in the future.

Effects of temperature on the respiratory metabolism,feeding and expression of three heat shock protein genes in Anadara broughtonii

Anadara broughtonii is one of the main commercially important species of shellfish in northern China.A.broughtonii lives in relatively stable subtidal zone where the clam could respond to environmental changes with minimum energy.Therefore,slight fluctuations in water environment may have a great impact on physiological processes such as feeding and metabolism.Large-scale mortality often occurs during the intermediate rearing processes,and high temperatures in summer are considered the leading cause of mortality.To understand the physiological and molecular response patterns of A.broughtonii at high temperatures and to estimate the appropriate metabolism temperature for A.broughtonii,the effects of temperature on the respiratory metabolism and food intake at different growth stages were studied.The response patterns of heat shock protein genes to heat stress and post-stress recovery were also explored.Results show that the temperature and growth stage of A.broughtonii were two important factors affecting metabolism and feeding.The optimum temperature for feeding and physiological activities in each shell-length group was 24℃.The temperature at which the peak metabolic rate occurred in each shell-length group increased with the increase in shell length.With the increase in heat stress,the expression of three heat shock protein genes(Abhsp60,Abhsp70,and Abhsp90)in the tissues of two size groups of A.broughtonii increased significantly when temperature was above 24℃and reached a peak at 30℃.After heat shock at 30℃for 2 h,the expression of the three heat shock protein genes rapidly increased,peaked at 2 h after the heat shock,and then gradually decreased to the level of the control group at 48 h after the heat shock.The three heat shock protein genes respond rapidly to heat stress and can be used as indicators to the cellular stress response in A.broughtonii under an environmental stress.

The polymorphism of heat shock protein 70 genes in Chinese Han population in Fujian province

To understand the polymorphism of the heat shock protein 70 （HSPTO） genes in Chinese Han population and to explore the co-relations between HSP70 polymorphism and disease, three polymorphic loci of HSP70 genes in 127 healthy Chinese Han population in Fujian province were analyzed by PCR and restriction enzyme analysis, and the genotypes and allele frequencies of HSPTO in different populations from various area were compared. It was found that the proportions of HSPTO-1 genotypes GG, GC and CC among Chinese Han population in Fujian province were 55.1%, 40.2% and 4.7% respectively, while those of HSP70-2 genotypes AA, AG and GG were 44.1%, 48.8 % and 6.9% respectively, and those of HSP70-hom genotypes TF, TC and CC were 59.8%, 37.0% and 3.2% respectively. The allele frequencies of G and C in HSP70-1 were 75.2% and 24.8% ; those of A and G in HSP70-2 were 68.5% and 31.5% and those ofT and C in HSP70-hom were 78.3% and 21.7% respectively. The distribution of the HSPTO-1 polymorphisms in Chinese Han population was almost the same as those in Japanese and Mexican populations, but it was rather different from those of American and Spanish populations with a significant differences. Meanwhile, the frequency of GG homozygote in HSPTO- 1 was signifi- cantly higher than those in American and Spanish populations. No significant difference was found in the distribution of HSPTO-2 polymorphism between Chinese and Japanese populations, in which the differences among American, Mexican and Spanish populations were quite obvious. The frequency of AA homozygote in HSPTO-2 was significantly higher than those in Mexican, American and Spanish populations, while, the distribution of HSPTO-hom genotype and allele frequency in Chinese Han population was almost just the same as those in Japanese and Mexican populations. Furthermore, it was also found that the genotype distribution and allele frequencies of the HSPTO genes in Han population of Fujian province were almost the same as those in Han population in Taiwan, but they were different in certain loci from those of Han population in Wuhan area. It is evident that the distribution of HSPTO gene polymorphisms among Chinese Han population are different from other regions in the world.

Establishment of a tetracycline-off and heat shock-on gene expression system in tobacco

The tetracycline （Tet）-off gene expression regulation system based on the TetR-VP16/Top10 construct has not been widely utilized in plants, for its highly expressed TetR-VP16 activator is toxic to some plants and repeatedly replenishing tetracycline to turn off the constitutively active system is a tedious process. To solve these problems, a Tet-off and heat shock （HS）-on gene expression regulation system was constructed in this study. This system is composed of a chimeric transactivator gene TetR-HSF that is derived from a Tet repressor （TetR） and a HS transcription factor （HSF） controlled by a HS promoter HSP70m, and a Tet operator containing hybrid promoter, Om35S, that drives expression of the β-glucuronidase （GUS） gene. The resultant system yields a GUS expression pattern similar to that of the HSP70m promoter under inducing temperatures and at 35 and 40℃ drives GUS expression to a similar level as the Cauliflower mosaic virus （CaMV） 35S promoter. Further examination revealed that the TetR-HSF and GUS genes were induced by HS, reaching peak expression after 1 and 6 h treatment, respectively, and the HS induction of the expression system could be inhibited by Tet. This system will provide a useful tool for transgenic studies of plants in the laboratory and in the field, including transgene function analysis, agronomic trait improvement, biopharmaceutical protein production and others.

Microarray-based Screening of Target Genes Regulated by Heat Shock Factor AtHsfA1a in Arabidopsis thaliana

[ Objective] This study aimed to screen target genes regulated by heat shock factor AtHsfAla in Arabidopsis thaliana. [ Method] Using AtHsfAla-in- serted mutant athsfala （SALK-068042） and wild-type A. thaliana seedlings as experimental materials, target genes regulated by heat shock factor AtHsfAla were screened by microarray assay. Differentially expressed genes were screened by multiple method. Specific functions of differentially expressed genes were analyzed by gene ontology （GO） analysis. Signal transduction pathways, in which differentia｜ly expressed genes were involved, were analyzed by pathway analysis. Gene-gene interaction network was constructed by Signal-Net. [ Result] A total of 3 672 differentially expressed genes were screened out. Up-regulated differentially expressed genes were involved in 198 functions and 7 signal transduction pathways; down-regulated differentially expressed genes were involved in 94 functions and 10 signal transduction pathways. In the signal transduction network, it was found that cwlNV4 and HXK3 had relatively high ability of mediation; AT1 G14240 and cwlNV4 ex- hibited the most interactions with other genes, which were located in key positions throughout the gene-gene interaction network. [ Conclusion] Heat shock factor AtHsfAla regulates a large number of target genes in A. thaliana.

THE ROLE OF INTRONIC HSE IN THE HEAT SHOCK INDUCED EXPRESSION OF HUMAN hsp 90β GENE

The first intron of human hsp90β gene is not only essential in maintaining high constitutive expression but also critical for heat shock inducibility of the hsp90β gene. Typical HSEs in the first intron play a vital role in the heat induced expression of human hsp90β gene. Slot blot analysis shows that hsp90β gene mRNA transcripts initiated from the 3’ of the first intron dominates over that of the first exon. The intronic HSEs of the hsp90β gene show much higher binding affinity toward the recombinant heat shock factor HSF1 than that of the recombinant heat shock factor HSF2.

Construction, Expression and Identification of a Recombinant BCG Vaccine Encoding Human Mycobacterium Tuberculosis Heat Shock Protein 65

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.

Influence of High Temperature on the Expression of Arabidopsis Heat Shock Transcription Factor AtHsfA1a

[ Objective] This study ~med to investigate the influence of high temperature on the expression of heat shock transcription factor AtHsfAla in different genotypes of Arabidopsis. [ Method ] Arabidopsis plants overexpressing heat shock transcription factor AtHsfA1 a were used as experimental materials and treated un- der high temperature at 39℃ for 1 rain and 5 min; total RNA of AtI-IsfAla was extracted, and the reverse transcription and amplification were conducted using RT- PCR technology, the amplification products were detected by electrophoresis. [ Result ] The expression levels of AtHsfA1 a in Arabidopsis plants overexpressing heat shock transcription factor AtHsfAla at high temperature and room temperature were higher than wild-type Arabidopsis; the expression levels of AtHsfAla in both wild-type Arab/dops/s and transgenic Arabidopsls plants overexpressing heat shock transcription factor AtHsfAla at high temperature of 39 ~C were higher than that at room temperature of 25 ~C, but the expression levels of AtHsfAla in wild-type Arab/dops/s and transgenic Arab/dops/s plants overexpressing heat shock transcription factor AtHsfAla varied little after high temperature treatment at 39 ~C for 1 rain or 5 rain. [ Conclusion] The expression of AtHsfAla is induced rapidly by high tem- perature, thus regulating the expression of early adversity-resistant genes. This study will lay the foundation demonstrating the mechanism of Arabidopsis heat shock transcription factor AtHsfAla.

Characterization of a TaJ Gene from Wheat

A novel J-domain protein gene was cloned from wheat （Triticum aestivum L.） using RT-PCR technology and named as TaJ. The J-domain protein is defined by the presence of a J-domain. The cDNA of T. aestivum gene, TaJ （GenBank accession number： DQ789026）, was 1263 bp and contained a complete open reading frame （ORF） encoding a J-domain protein of 420 amino acid residues. The predicted amino acid sequence of TaJ possesses three functionally essential domains： the Nterminal J-domain which includes the highly conserved HPD tripeptide, an adjacent domain that is rich in glycine and phenylalanine residues （G/F） and a Cysteine-rich zinc-finger domain with four repeats of CxxCxGxG that is important for protein interactions. The C-terminal of TaJ was -CAQQ, a farnesylation motif. The full-length deduced amino acid sequence of TaJ is highly homologous to J-domain proteins from various plant species. Southern blot analysis indicated that a single copy of TaJ existed in wheat genome. The expression pattern of TaJ performed by real-time PCR demonstrated that heat shock （HS） at 37℃ induced the expression of TaJ rapidly and strongly, but the response of the TaJ gene to cold stress was much slower than that to HS. Tissue-specific expression analysis showed that the expression level of TaJ gene was much higher in leaves than that in roots.

Bioinformatics analysis of microarray data to explore the key genes involved in HSF4 mutation-induced cataract

AIM： To reveal the mechanisms of heat-shock transcription factor 4 （HSF4） mutation-induced cataract.METHODS： GSE22362, including 3 HSF4-null lens and 3 wild-type lens, was obtained from Gene Expression Omnibus database. After data preprocessing, the differentially expressed genes （DEGs） were identified using the limma package. Based on Database for Annotation, Visualization and Integrated Discovery （DAVID） tool, functional and pathway enrichment analyses were performed for the DEGs. Followed by protein-protein interaction （PPI） network was constructed using STRING database and Cytoscape software. Furthermore, the validated microRNA （miRNA）-DEG pairs were obtained from miRWalk2.0 database, and then miRNA-DEG regulatory network was visualized by Cytoscape software. RESULTS： A total of 176 DEGs were identified in HSF4-null lens compared with wild-type lens. In the PPI network, FBJ osteosarcoma oncogene （FOS）, early growth response 1 （EGR1） and heme oxygenase （decycling） 1 （HMOX1） had higher degrees and could interact with each other. Besides, mmu-miR-15a-5p and mmu-miR-26a-5p were among the top 10 miRNAs in the miRNA-DEG regulatory network. Additionally, mmu-miR-26a-5p could target EGR1 in the regulatory network. CONCLUSION： FOS, EGR1, HMOX1, mmu-miR-26a-5p and mmu-miR-15a-5p might function in the pathogenesis of HSF4 mutation-induced cataract.

Constitutively Expressed αB—Crystallin in Heat Schock Transcription Factor 1 Knockout Mice Myocardium

Objective-To investigate the effects of heat shock transcription factor 1) gene on the constitutivety expressed αB-CrystaUin (aBC) in mice myocardium. Methods-The expression levels of constitutive aBC in HSF1 knockout (hsf1 - /- ) and HSFl wild type (As/1 + /+) mice myocardium were evaluated by western blot and immunohistochemistry. Results : The αBC levels in hsfl -/- and hsfl +/+ were 68. 42±4. 16, 100. 00±7. 58, respectively (P<0. 05, cytoso-lic fraction) , and 20. 53±1. 01, 37. 55±1. 91, respectively (P<0. 05, pellet fraction). The aBC signals decreased significantly in hsfl -/- myocardium when compared with those in hsfl +/+ myocardium stained with fluorescence immunohistochemistry. Conclusion-HSF1 is an important, but not the only factor, which mediates the constitutively expressed aBC.

半滑舌鳎响应急性高温胁迫的生理变化和相关基因表达

夏季高温会引起半滑舌鳎(Cynoglossus semilaevis)的应激,甚至造成死亡,是工厂化养殖的重要影响因素之一。为探究高温胁迫对半滑舌鳎肝脏氧化损伤及热应激相关基因的影响,本研究选取半滑舌鳎一个全同胞家系为实验对象,通过连续升温达到高温胁迫条件(35℃)后,分别在0、3、6、12和24 h采集肝脏组织,进行苏木精–伊红染色法(HE)和TUNEL染色并观察细胞损伤情况,测定抗氧化酶活性及丙二醛(MDA)含量并检测应激相关基因heatshockprotein familyAmember1A(hspa1a)、heatshockprotein90betafamilymember1(hsp90b1)和dual-specificity phosphatase 1(dusp1)的表达变化。结果显示,急性高温胁迫会造成半滑舌鳎肝脏组织发生明显病理变化并出现细胞凋亡;超氧化物歧化酶(SOD)活性在高温胁迫6 h时显著高于对照组(P<0.05),谷胱甘肽过氧化物酶(GPx)活性在高温胁迫12h时显著高于对照组(P<0.05),过氧化氢酶(CAT)活性在高温胁迫0h时显著高于对照组(P<0.05),MDA含量在高温胁迫24h时显著高于对照组(P<0.05);热休克蛋白基因hspa1a和hsp90b1分别在高温胁迫0 h和3 h时显著上调表达,热应激相关基因dusp1在高温胁迫3h时显著上调表达。综上所述,急性高温胁迫下,半滑舌鳎肝脏发生氧化应激,短期内机体可调动抗氧化系统加速清除活性氧,并激活热应激相关基因表达。该研究可为解析半滑舌鳎对高温胁迫的响应机制、预防夏季高温大规模死亡的发生以及开展耐高温良种的选育等提供参考。

CD8 T cell response in a phase I study of therapeutic vaccination of advanced NSCLC with allogeneic tumor cells secreting endoplasmic reticulum-chaperone gp96-Ig-peptide complexes

Antigen containing, allogeneic cells secreting the genetically modified protein and peptide-chaperone gp96-Ig cross, prime and expand antigen specific CD8 T cells with therapeutic antitumor activity in mice. In a first in man phase I study, we now report the results of therapeutic vaccination of non-small cell lung cancer (NSCLC) patients with an established, allogeneic non-small cell lung adenocarcinoma cell line secreting gp96-Ig. Advanced NSCLC-patients stage IIIB or IV of any histological subtype were enrolled and treated with up to 36 vaccinations over the course of 18 weeks. Primary endpoint was safety, secondary endpoints tumor response and overall survival. Measurement of tumor antigen specific CD8 CTL responses is precluded by the lack of known NSCLC associated antigens. Therefore, we measured patient CD8 T cell-IFN-γ responses to allo-antigens of the vaccine cells as surrogate for tumor antigen specific CD8 CTL. In 7 of 18 treated patients tumor growth was stabilized, however none of the 18 patients had an objective tumor response by RECIST criteria. Of 15 patients evaluable for immune response, 11 responded to vaccination with more than twofold increase in CD8-IFN-γ frequency above baseline. These patients had a median survival time of 16.5 months. Four patients who had no CD8 response above base line had survival times from 2.1 to 6.7 months. Our data are consistent with the concept that generation of CD8 CTL by therapeutic vaccination may delay tumor growth and progression and mediate prolonged survival even in the absence of objective tumor responses. Further studies will be required to test this concept and promising result.

转录组测序分析外源水杨酸诱导茶树热激蛋白基因的响应

水杨酸是诱导植物抗性机制中重要的信号分子,外源喷施水杨酸能够调控多种防御相关蛋白质,提升农作物的抗病能力。开展外源水杨酸诱导茶树抗性机制的研究能够挖掘抗病基因,为茶树抗病育种提供分子基础。本研究采集外源喷施水杨酸0 h、6 h、12 h、24 h、48 h的茶树叶片进行转录组测序与分析,结果表明,外源喷施水杨酸6 h、12 h、24 h、48 h时茶树叶片内差异表达基因数量分别为9 360个、3 399个、596个、115个,外源水杨酸处理后各时间点均发生差异表达的基因共604个。KEGG功能富集结果显示,处理后6 h时富集于植物激素信号转导、植物-病原菌互作、核糖体、剪接体和碳代谢通路上的差异表达基因数量分别为95个、73个、121个、94个、154个。差异表达基因中有12个热激因子基因、40个热激蛋白基因和12个WRKY家族转录因子基因上调表达。处理48 h后,无上调表达的热激因子基因,但仍有28个热激蛋白基因上调表达。病程相关蛋白基因在检测阶段均上调表达。外源水杨酸的诱导作用在处理6 h时最为明显,并且引起了大量热激蛋白的响应。本研究结果为开展外源水杨酸诱导茶树抗病机制和茶树抗病分子育种研究提供了参考。

急性温度和盐度胁迫对罗氏沼虾热休克蛋白基因表达及抗氧化酶活性的影响

【目的】探究急性温度和盐度胁迫对罗氏沼虾热休克蛋白基因及抗氧化酶活性的影响,为实际生产中罗氏沼虾的健康养殖提供理论依据。【方法】采用双因素试验,设不同温度(20、24、28和32℃)和盐度(0.5‰、4.0‰、8.0‰和12.0‰),共16个处理组,将罗氏沼虾从初始温度(24℃)和盐度(0.5‰)环境转移至各处理组环境中进行急性胁迫,分别在胁迫24和48 h时采集罗氏沼虾鳃组织及肝胰腺组织,测定热休克蛋白70基因(HSP70)相对表达量、超氧化物歧化酶(SOD)活性和谷胱甘肽过氧化物酶(GPX)活性。【结果】胁迫24 h时,在相同温度下,随着盐度的升高,罗氏沼虾鳃组织和肝胰腺组织HSP70基因相对表达量均整体呈先上升后下降的变化趋势;在0.5‰和4.0‰盐度下,温度为20、28和32℃时罗氏沼虾鳃组织和肝胰腺组织HSP70基因相对表达量均高于24℃,且肝胰腺组织HSP70基因相对表达量在温度为28和32℃时显著高于20℃(P<0.05,下同)。胁迫48 h后,温度为24℃时,8.0‰和12.0‰盐度下罗氏沼虾鳃组织及肝胰腺组织HSP70基因相对表达量均显著高于0.5‰与4.0‰盐度。胁迫24 h时,在相同温度下,随着盐度的升高,罗氏沼虾腮组织SOD和GPX活性均整体呈上升趋势;温度为24℃时,随着盐度升高,肝胰腺组织SOD和GPX活性均无显著变化(P>0.05),而在其他温度下出现明显变化;在相同盐度下,随着温度的升高,罗氏沼虾鳃组织SOD和肝胰腺组织GPX活性均整体呈先下降后上升的变化趋势,腮组织GPX活性和肝胰腺组织SOD活性均整体呈上升趋势。双因素方差分析结果表明,温度、盐度及其交互作用对罗氏沼虾鳃组织和肝胰腺组织HSP70基因表达与抗氧化酶(SOD和GPX)活性均有极显著影响(P<0.01)。【结论】急性温度和盐度胁迫会引起罗氏沼虾HSP70基因相对表达量升高,且温度骤升较温度骤降更能诱导HSP70基因表达。温度和盐度骤变能引起罗氏沼虾抗氧化酶活性变化,且温度对抗氧化酶活性的影响大于盐度。罗氏沼虾通过调控HSP70基因相对表达量、SOD活性及GPX活性协同调节生理机能以适应外部盐度和温度变化,增强对恶劣环境的耐受性。

The regulatory effect of memantine on expression and synthesis of heat shock protein 70 gene in neonatal rat models with cerebral hypoxic ischemia

To evaluate the neuroprotective effect of memantine, a non-competitive antagonist at the N-methyl-D-aspartate receptor, against hypoxic ischemia (HI) by exploring its regulation on the expression and synthesis of heat shock protein 70 (HSP70) gene in neonatal rat models with cerebral HI Methods Memantine was intraperitoneally injected at a dose of 20 mg/kg in neonatal rat models either before (PRE group) or after (POST group) induction of HI The expression and synthesis of the HSP70 gene and its corresponding product were determined by rapid competitive PCR and immunohistochemistry, respectively Results There was an increase in the expression of HSP70 mRNA two hours after induction of HI, which reached its peak at 48 hours, then decreased gradually The same expression occurred at relatively low levels in the control group Also, HSP70 synthesis was detected as early as 2h after HI, reached its peak between 48 and 72 hours, then declined over time After memantine administration, the expression of the gene and its synthesis of the corresponding product decreased significantly during the time intervals 24-72 h for the gene and 48-72 h for the product compared to the HI group Conclusion It was shown that HI is very sensitive to the expression of the HSP70 gene and synthesis of its corresponding product, which could be regulated by memantine The latter may have the ability to reduce brain damage; thus decreased HSP70 mRNA expression could be a marker for HI It is suggested that memantine can be a promising agent for neuroprotection against HI, although an overall and abstract assessment of memantine is required to see if it can be used on neonates clinically later on

Cloning of heat shock protein genes from the brown planthopper, Nilaparvata lugens, and the small brown planthopper, Laodelphax striatellus, and their expression in relation to thermal stress

Three heat shock protein （HSP） genes （hsp70, hsc70, hsp90） were partially cloned from the brown planthopper Nilaparvata lugens and the small brown planthopper Laodelphax striatellus （Homoptera： Delphacidae）, which are serious pests of the rice plant. Sequence comparisons at the deduced amino acid level showed that the three HSPs of planthoppers were most homologous to corresponding HSPs of dipteran and lepidopteran species. Identities of both heat shock cognate 70 and HSP90 were higher than HSP70 in both species. Identity of the HSP70 between the two planthopper species was only 81%, a value much lower than seen among fly and moth groups. Effects of heat and cold shocks were demonstrated on expression of the three hsp genes in the two planthopper species. Heat shock （40 ℃） upregulated the hsp90 level but did not change the hsc70 level in either the nymph and adult stages of either species. On the other hand, the hsp70 level was only upregulated in L. striatellus. This heat shock response was prompt and lasted only for 1 h after treatment. In contrast, cold shock at 4℃ did not change the expression levels of any hsp in either species.

Rapid Identification of Mycobacterium Leprae by Polymerase Chain Reaction-restriction Fragment Length Polymorphism Analysis of the Heat Shock Protein 65 Gene from Skin Specimens

INTRODUCTIONLeprosy caused by Mvcobacterium leprae （M. leprae）, is a chronic granulomatous disease affecting the skin and peripheral nervous system, which is transmitted through direct contact with nontreated or inadequate treatment patients. Diagnosis of leprosy depends on the clinical signs and symptoms and slit skin smear positivity. However, it＇s sometimes similar with other granulomatous disease caused by mycobacterial infection. Early stage leprosy is difficult to diagnose by clinical criterion alone because the sensitivity of acid-fast bacilli staining is quite low. The polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） shows the great advantage in rapid identification and diagnosis for early cases and has a differentiation between leprosy and nonleprosy cases.