In vivo Gonadoprotective Effects of Myricetin on Cisplatin- Induced Testicular Damage via Suppression of TLR4/NF-kB Inflammation Pathway and Heat-Shock Response / Efectos Gonadoprotectores in vivo de la Miricetina sobre el Daño Testicular Inducido por Cisplatino Mediante la Supresión de la Vía de Inflamación TLR4/NF-kB y la Respuesta al Choque Térmico

SUMMARY: The aim of this study is to reveal the gonadoprotective effects of myricetin (MYC), which has many biological properties, on cisplatin (CP)-induced testicular damage in rats. For this purpose, 40 male Wistar albino rats were divided into 4 groups as Control (group given no treatment), MYC (group given 5 mg/kg/i.p myricetin for 7 days), CP (group given 7 mg/kg/i.p cisplatin at 7th day) and MYC + CP (group given 5 mg/kg/i.p myricetin for 7 days before 7 mg/kg/i.p cisplatin injection). After administrations, testicular tissues of animals were extracted and processed according to tissue processing protocol. Hematoxylin & Eosin staining were performed to evaluate the histopathological changes and Johnsen'sTesticular Biopsy Score (JTBS) was applied and mean seminiferous tubule diameters (MSTD) were measured to compare experimental groups in terms of histopathological changes. Moreover, TLR4, NF-kB, HSP70 and HSP90 expression levels were detected by immunohistochemical staining and the density of immunoreactivity were measured to determine the difference in the expression levels of these factors among groups. Additionally, testicular apoptosis was detected via TUNEL assay. JTBS and MSTD data were significantly lower in CP group compared to other groups and MYC administrations significantly protects testicular tissue against CP-induced damage. Moreover, TLR4, NF-kB, HSP70 and HSP90 expressions and apoptotic cells significantly increased in the CP group (p<0.05). However, MYC administrations exerted a strong gonadoprotective effect on testicular tissue in terms of these parameters in MYC+CP group (p<0.05). According to our results, we suggested that MYC can be considered as a protective agent against cisplatin-induced testicular damage.

El objetivo de este estudio es revelar los efectos gonadoprotectores de la miricetina (MYC), que tiene muchas propiedades biológicas, sobre el daño testicular inducido por cisplatino (CP) en ratas. Para este propósito, se dividieron 40 ratas albinas Wistar macho en 4 grupos: Control (grupo que no recibió tratamiento), MYC (grupo que recibió 5 mg/kg/i.p de miricetina durante 7 días), CP (grupo que recibió 7 mg/kg/i.p de cisplatino al séptimo día) y MYC + CP (grupo que recibió 5 mg/ kg/i.p de miricetina durante 7 días antes de la inyección de 7 mg/ kg/i.p de cisplatino). Después de las administraciones, se extrajeron y procesaron tejidos testiculares de animales según el protocolo de procesamiento de tejidos. Se realizó tinción con hematoxilina y eosina para evaluar los cambios histopatológicos y se aplicó la puntuación de biopsia testicular de Johnsen (JTBS) y se midieron los diámetros medios de los túbulos seminíferos (MSTD) para comparar los grupos experimentales en términos de cambios histopatológicos. Además, los niveles de expresión de TLR4, NF-kB, HSP70 y HSP90 se detectaron mediante tinción inmunohistoquímica y se midió la densidad de inmunorreactividad para determinar la diferencia en los niveles de expresión de estos factores entre los grupos. Además, se detectó apoptosis testicular mediante el ensayo TUNEL. Los datos de JTBS y MSTD fueron significativamente más bajos en el grupo CP en comparación con otros grupos y las administraciones de MYC protegen significativamente el tejido testicular contra el daño inducido por CP. Además, las expresiones de TLR4, NF-kB, HSP70 y HSP90 y las células apoptóticas aumentaron significativamente en el grupo CP (p<0,05). Sin embargo, las administraciones de MYC ejercieron un fuerte efecto gonadoprotector sobre el tejido testicular en términos de estos parámetros en el grupo MYC+CP (p<0,05). Según nuestros resultados, sugerimos que MYC puede considerarse como un agente protector contra el daño testicular inducido por cisplatino.

Heat shock response in noise-induced hearing loss: effects of alanyl-glutamine dipeptide supplementation on heat shock proteins status / Resposta de choque térmico em perda auditiva induzida por ruído: efeitos da suplementação com dipeptídeos de alanil-glutamina sobre o estado das proteínas de choque térmico

Abstract Introduction: The 72 kDa heat shock protein, HSP72, located intracellularly provides cochlear cytoprotective and anti-inflammatory roles in the inner ear during stressful noise challenges. The expression of intracellular HSP72 (iHSP72) can be potentiated by alanyl-glutamine dipeptide supplementation. Conversely, these proteins act as pro-inflammatory signals in the extracellular milieu (eHSP72). Objective: We explore whether noise-induced hearing loss promotes both intracellular and extracellular HSP72 heat shock response alterations, and if alanyl-glutamine dipeptide supplementation could modify heat shock response and prevent hearing loss. Methods: Female 90 day-old Wistar rats (n = 32) were randomly divided into four groups: control, noise-induced hearing loss, treated with alanyl-glutamine dipeptide and noise-induced hearing loss plus alanyl-glutamine dipeptide. Auditory brainstem responses were evaluated before noise exposure (124 dB SPL for 2 h) and 14 days after. Cochlea, nuclear cochlear complex and plasma samples were collected for the measurement of intracellular HSP72 and extracellular HSP72 by a high-sensitivity ELISA kit. Results: We found an increase in both iHSP72 and eHSP72 levels in the noise-induced hearing loss group, which was alleviated by alanyl-glutamine dipeptide treatment. Furthermore, H-index of HSP72 (plasma/cochlea eHSP72/iHSP72 ratio) was increased in the noise-induced hearing loss group, but prevented by alanyl-glutamine dipeptide treatment, although alanyl-glutamine dipeptide had no effect on auditory threshold. Conclusions: Our data indicates that cochlear damage induced by noise exposure is accompanied by local and systemic heat shock response markers. Also, alanyl-glutamine reduced stress markers even though it had no effect on noise-induced hearing loss. Finally, plasma levels of 72 kDa heat shock proteins can be used as a biomarker of auditory stress after noise exposure.

Resumo Introdução: A proteína de choque térmico de 72 kDa, HSP72 localizada intracelularmente, tem papéis citoprotetores e anti-inflamatórios cocleares na orelha interna durante situações de ruído estressantes. A expressão dessa proteína pode ser potencializada pela suplementação com dipeptídeo de alanil-glutamina. Por outro lado, essas proteínas atuam como sinais pró-inflamatórios no meio extracelular. Objetivo: Investigar se a perda auditiva induzida por ruído promove alterações tanto das proteínas HSP72 intracelulares quanto extracelulares na resposta de choque térmico e se a suplementação com alanil-glutamina pode modificar a resposta de choque térmico e evitar a perda auditiva. Método: Ratos Wistar fêmeas, com 90 dias de idade (n = 32), foram divididos aleatoriamente em quatro grupos: controle, perda auditiva induzida por ruído, tratados com alanil-glutamina e perda auditiva induzida por ruído mais alanil-glutamina. Os potenciais evocados auditivos do tronco encefálico foram avaliados antes da exposição ao ruído (124 dB NPS por 2 h) e 14 dias após. A cóclea, o complexo nuclear coclear e amostras de plasma foram coletadas para mensuração de HSP72 intra e extracelular com um kit Elisa de alta sensibilidade. Resultados: Houve um aumento nos níveis de HSP72 intra e extracelular no grupo perda auditiva induzida por ruído, que foi minimizado pelo tratamento com alanil-glutamina. Além disso, o índice H das HSP72 (razão HSP72 extracelular/HSP72intracelular plasma/cóclea) aumentou no grupo perda auditiva induzida por ruído, mas foi limitado pelo tratamento com alanil-glutamina, embora o alanil-glutamina não tenha efeito no limiar auditivo. Conclusões: Nossos dados indicam que o dano coclear induzido pela exposição ao ruído é acompanhado por marcadores da resposta de choque térmico locais e sistêmicos. Além disso, alanil-glutamina reduziu os marcadores de estresse, mesmo não tendo efeito sobre a perda auditiva induzida por ruído. Finalmente, os níveis plasmáticos de proteínas de choque térmico de 72 kDa podem ser usados como biomarcador do estresse auditivo, após a exposição ao ruído.

Análise dos efeitos biológicos associados ao fator de choque térmico 1 (HSF1) no carcinoma de células escamosas oral / Analysis of the biological effects associated with heat shock factor 1 (HSF1) in oral squamous cell carcinoma

O carcinoma de células escamosas oral exibe altas taxas de morbimortalidade e evidências em vários tipos tumorais mostram que processos associados à iniciação, à progressão e à resistência terapêutica são regulados por HSF1. Portanto, esclarecer as vias de participação de HSF1 no câncer oral pode auxiliar no entendimento do seu comportamento biológico. Em uma pesquisa previamente desenvolvida por nosso grupo, foram realizados a análise clinicopatológica e o estudo da imunoexpressão de HSF1 em 70 casos de carcinoma de células escamosas de língua oral (CCELO) em comparação com 30 espécimes de mucosa oral normal (MON). Nesta atual investigação, avaliou-se a participação de HSF1 na tumorigênese do CCELO, através de experimentos in vitro com a linhagem celular SCC15, silenciada e não silenciada, com silenciamento confirmado por qRT-PCR e Western Blot. Foram analisadas a viabilidade e proliferação celular, (CellTiter e BRDU, respectivamente), influência no ciclo celular (iodeto de propideo e análise por citometria de fluxo), capacidade de invasão (sistema transwell/Matrigel)e transição epitélio-mesenquimal (TEM) (expressão de E-caderina e vimentina por qRT-PCR). Nossos resultados anteriores evidenciaram que quanto aos casos de CCELO, 57,1% exibiram estadiamento clínico III ou IV, 82,9% foram gradados como de alto grau segundo Bryne (1998), 47,1% como de alto risco segundo Brandwein-Gensler et al. (2005) e 58,8% como de alto risco de acordo com o modelo BD. Observou-se repercussão da gradação de Bryne (1998) (p= 0,05) na sobrevida livre de doença. Tamanho do tumor T3 ou T4 (p= 0,04), recidiva local (p= 0,02) e modelo BD (p=0,02) repercutiram na sobrevida global. Encontrou-se previamente resultado significativo (p<0,01) quando se comparou a imunoexpressão de HSF1 entre a MON e o CCELO, sem associações significativas da imunoexpressão com os parâmetros clinicopatológicos. A partir dos estudos funcionais, observou-se que HSF1 é superexpresso na linhagem SCC15 comparada aos queratinócitos imortalizados (p<0,005) e que o silenciamento deste gene inibiu a proliferação celular (p< 0,005), avanço nas fases do ciclo celular, com aumento do número de células nas fases G0/G1 (p<0,01) e redução das células na fase S (p<0,001), capacidade de invasão (p<0,05) e TEM, com diminuição da expressão de vimentina (p<0,001) e aumento de E-caderina (p<0,05), quando comparadas as linhagens silenciada e controle. Diante destes resultados, sugere-se que HSF1 pode desempenhar diversas funções que ajudam a manter a estabilidade celular em meio às condições estressoras do microambiente tumoral. Assim, futuramente, estratégias envolvendo sua regulação pode ser uma terapia útil no controle da progressão do câncer oral (AU).

Oral squamous cell carcinoma exhibits high rates of morbimortality and evidence in several tumor types shows that processes associated with initiation, progression and therapeutic resistance are regulated by HSF1. Therefore, to clarify the pathways of HSF1 participation in the oral cancer may help in the understanding of its biological behavior. In research previously developed by our group, a clinicopathological analysis and an immunoexpression study of HSF1 of 70 cases of oral tongue squamous cell carcinoma (OTSCC) were performed in comparison with 30 samples of the normal oral mucosa (NOM). In this current investigation, the role of HSF1 in OTSCC tumorigenesis was evaluated, through in vitro experiments with the SCC15 cell line, silenced and non-silenced, with silencing confirmed by qRT-PCR and Western Blot. Cell viability and proliferation (CellTiter and BrdU, respectively), influence on cell cycle (propidium iodide and flow cytometry analysis), invasion capacity (transwell / Matrigel system), and epithelial-mesenchymal (EMT) (expression of E-cadherin and vimentin by qRT-PCR) were evaluated. Our previous results showed that as for the cases of OTSCC, 57.1% exhibited clinical stage III or IV, 82.9% were graded as high grade according to Bryne (1998), 47.1% as high risk according to BrandweinGensler et al. (2005) and 58.8% as high risk according to the BD model. Bryne's gradation (1998) (p = 0.05) had an impact on disease-free survival. Tumor size T3 or T4 (p = 0.04), local recurrence (p = 0.02) and BD model (p = 0.02) impacted overall survival. A significant initial result (p <0.01) was found when comparing an HSF1 immunoexpression between NOM and OTSCC, with no significant association of immunoexpression with clinicopathological tests. From the functional studies, it was observed that HSF1 is overexpressed in the SCC15 cell line compared to immortalized keratinocytes (p <0.005) and that the silencing of this gene inhibited cell proliferation (p <0.005), advance in the cell cycle phases, with an increase in the number of cells in phases G0/G1 (p <0.01) and reduction of cells in phase S (p <0.001), invasion capacity (p <0.05) and EMT, with decreased vimentin expression (p <0.001) and increased E-cadherin (p <0.05), when compared to silenced and control lines. Given these results, it is suggested that HSF1 can exert a range of functions that maintain cell stability amid the stressful conditions of the tumor microenvironment. Thus, in the future, strategies involving its regulation may be a useful therapeutic tool in controlling the progress of the oral cancer (AU).

Heat shock proteins: protection and potential biomarkers for ischemic injury of cardiomyocytes after surgery

Abstract The heat shock proteins are endogenous proteins with the ability to act as molecular chaperones. Methods that provide cell protection by way of some damage can positively influence the results of surgery. The present review summarizes current knowledge concerning the cardioprotective role of the heat shock proteins as occurs in heart damage, including relevant information about the stresses that regulate the expression of these proteins and their potential role as biomarkers of heart disease.

Physiological regulation of the heat shock response by glutamine: implications for chronic low-grade inflammatory diseases in age-related conditions

Aging is an intricate process modulated by different molecular and cellular events, such as genome instability, epigenetic and transcriptional changes, molecular damage, cell death and senescence, inflammation, and metabolic dysfunction. Particularly, protein quality control (chaperone systems) tends to be negatively affected by aging, thus leading to cellular senescence in metabolic tissues and, as a consequence, to the increasing dissemination of inflammation throughout the body. The heat shock (HS) response and its associated expression of the 70 kDa family of heat shock proteins (HSP70),which are anti-inflammatory molecular chaperones, are found to be markedly decreased during muscle inactivity and aging, while evidence supports the loss of HSP70 as a key mechanism which may drive muscle atrophy, contractile dysfunction, and reduced regenerative capacity. In addition, abnormal stress response is linked with higher incidence of neurodegenerative diseases as well as low-grade inflammatory diseases that are associated with physical inactivity and obesity. Therefore, strategies to increase or, at least, to maintain the levels of HSP70, and its accompanying HS response to stress, are key to reduce biological cell dysfunctions that occur in aging. In this sense, physical exercise is of note as it is the most powerful inducer of the HS response, comparable only to heat stress and fever-like conditions. On the other hand, the amino acidL-glutamine, whose production within the skeletal muscle and liberation into the bloodstream is dependent on muscle activity, is a potentializer of HSP70 expression and HS response, particularly via its entering in hexosamine biosynthetic pathway (HBP). Herein, we discuss the collaborative role of glutamine (and its donors/precursors) and physical exercise (mostly responsible for glutamine release into the circulation) as potential tools to increase HSP70 expression and the HS response in the elderly.

Molecular structure and alternative splicing analysis of heat shock factors of Schistosoma japonicum / 中国血吸虫病防治杂志

Objective To clone and identify the heat shock factors(HSFs)of Schistosoma japonicum and analyze its molec?ular structure and alternative splicing pattern. Methods The New Zealand rabbits were infected with the cercariae of Schistoso?ma japonicum and were killed and dissected 42 days post?infection,and the adult worms of S. japonicum and the livers of the rabbits were harvested. Then,the total RNA was extracted by using Trizol reagent. The Sj?hsf open reading frame(ORF)and the alternative splicing fragments were amplified by RT?PCR from the female,male and egg samples,then cloned and verified by enzyme digestion and sequencing. DNAMAN 8.0,InterPro,Mega 6 combined with the Internet databases were utilized to clarify the gene structure,functional domains,alternative splicing pattern,and the homology and phylogenetic tree of HSFs. Re?sults Sj?hsf ORF and the alternative splicing fragments were amplified from the female,male and egg samples of S. japonicum by RT?PCR. After cloning,the positive recombinant plasmids pBSjHSFf?F,pBSjHSFf?M,pBSjHSFf?E containing Sj?hsf ORF, pBSjHSFs?F,pBSjHSFs?M,pBSjHSFs?E with Sj?hsf alternative splicing fragments were identified by enzyme digestion and se?quencing. Three alternative splicing Sj?hsf isoforms were observed through sequence analysis:Sj?hsf?isoform1(2 050 bp),Sj?hsf ?isoform2(2 086 bp)and Sj?hsf?isoform3(2 111 bp);the GenBank accession numbers were KU954546,KX119143 and KX119144,respectively. All the three isoforms located in the same Contig SJC_S000780 of S. japonicum genome and all ex?pressed at female,male and egg stages,but Sj?hsf?isoform1 with a high?level expression. Sj?HSF?isoform1(671 aa)and Sj?HSF?isoform2(683 aa)had DBD(DNA binding domain),HR?A/B and HR?C domains,while Sj?HSF?isoform3(282 aa)stopped in advance without HR?C domain. Phylogenetic tree analysis of HSFs illustrated that Sj?HSFs belonged to HSF1 family,with a close phylogenetic relationship to Sm?HSFs. Conclusions There are three alternative splicing isoforms of Sj?HSF existing in the female,male and egg stages of S. japonicum,but Sj?HSF?isoform1 expresses in a high?level. This study lays the foundation for further study on molecular mechanisms of Sj?HSFs in regulating the heat shock response system.

Heat shock improves Sca-1+ cell transplantation for treatment of myocardial infarction in mice / 中国组织工程研究

BACKGROUND:Recent studies have found that stem cels can directly differentiate into mature myocardial cels or promote their regeneration, providing a new therapeutic strategy for the treatment of myocardial infarction. However, the low cel transplantation rate reduces the myocardial differentiation ability and myocardial repair. OBJECTIVE: To study the role of heat shock treatment in Sca-1+ cel transplantation for treatment of myocardial infarction in mice. METHODS:Sca-1+ cels were isolated from the bone marrow of mice using magnetic bead sorting method, and were subjected to heat shock treatment. Animal models of myocardial infarction were made in mice, and then randomized into two groups: heat shock group and non-heat shock group, which were given 1 mL heat shock-treated Sca-1+ cels and 1 mL non-heat shock-treated Sca-1+ celsvia the tail vein, respectively. After transplantation, cel survival, heart function, myocardial cel apoptosis and myocardial fibrosis were detected. Meanwhile, the expressions of heat shock factor (HSP), HSP70 and miR-34a in the left ventricle were measured. RESULTS AND CONCLUSION:(1) The expression of sry gene in the heat shock group was significantly higher than that in the non-heat shock group. (2) The left ventricular ejection fraction and fractional shortening in the heat shock group were significantly higher than those in the non-heat shock group. The left ventricular end-diastolic diameter and systolic diameter in the heat shock group were significantly lower than those in the non-heat shock group. (3) The cardiac fibrosis and myocardial cel apoptosis in the heat shock group were significantly lower than those in the non-heat shock group. (4) The HSF and HSP70 expression in the left ventricle was significantly higher in the heat shock group than the non-heat shock group, and the miR-34a expression in the left ventricle was significantly lower in the heat shock group than the non-heat shock group. These findings indicate that heat shock-treated Sca-1+ cel transplantation can reduce myocardial apoptosis and infarct size, and improve heart function of mice with myocardial infarction.

The Role of Heat Shock Response in Insulin Resistance and Diabetes

The expansion of life-style related diseases, such as metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), appears to be unstoppable. It is also difficult to cease their complications in spite of many antidiabetic medications or intervention of public administration. We and our collaborators found that physical medicine using simultaneous stimulation of heat with mild electric current activates heat shock response, thereby reducing visceral adiposity, insulin resistance, chronic inflammation and improving glucose homeostasis in mice models of T2DM, as well as in humans with MS or T2DM. This combination therapy exerts novel action on insulin signaling, beta-cell protection and body compositions, and may provide a new therapeutic alternative in diabetic treatment strategy.

Effect of high temperature preconditioning on hydrogen peroxide-induced expression of mitochondrial metallothionein in rat cardiomyocytes / 中华麻醉学杂志

Objective To investigate the effects of high temperature preconditioning on hydrogen peroxide (H2 O2)-induced expression of mitochondrial metallothionein (MT) in rat cardiomyocytes.Methods The rat cardiomyocytes H9C2 cultured in vitro were randomly divided into 3 groups (n =6 each):control group (group C) ;H2O2 group (group H2O2); high temperature preconditioning group (group HTP).The cells were continuously cultured for 3 h in group C.The cells were cultured for 3 h in serum-free DMEM liquid culture medium containing H2O2 0.5 mmol/L in an incubator filled with 5％ CO2 at 37 ℃ in group H2O2.In group HTP,the cells were cultured in serum-containing DMEM liquid culture medium,then placed in a warm bath of 42 ℃ for 1 h,cultured for 12 h in an incubator filled with 5％ CO2 at 37 ℃,DMEM liquid culture medium was then removed,and the other procedures were similar to those previously described in group H2 O2.Myocardial cell apoptosis was observed by flow cytometry.The apoptotic rate was calculated.The ultrastructure of myocardial mitochondria was examined with electron microscope.The expression of mitochondrial MT in cardiomyocytes was determined using Western blot.Results Compared with group C,the apoptotic rate was significantly increased,and the expression of mitochondrial MT was up-regulated in groups H2O2 and HTP (P ＜ 0.01).The apoptotic rate was significantly lower,and the expression of mitochondrial MT was higher in group HTP than in group H2O2 (P ＜ 0.01).The mitochondrial injury was attenuated in group HTP as compared with group H2 O2.Conclusion The mechanism by which high temperature preconditioning reduces H2 O2-induced myocardial damage may be related to up-regulation of expression of mitochondrial MT in cardiomyocytes and endogenous myocardium-protective mechanism in rats.

The effects of heat and osmotic stress on vibrio parahaemolyticus survival rates, cellular morphology and TDH production.

Vibrio parahaemolyticus is an emerging enteric pathogen often associated with the consumption of improperly cooked seafood. The major virulence factor of this pathogen is represented by the termostable direct hemolysin (TDH). When entering the host organism, the vibrion has to face a series of environmental changes (such as temperature and osmolarity) which determine various cellular responses among which the heat shock response has an important role. The purpose of this study was to analyze the survival profiles, the morphological changes and the TDH production in V. parahaemolyticus strains grown under sub-lethal heat and osmotic stress. Our study has demonstrated the thermolotolerance in V. parahaemolyticus strains and the absence of cross-protection between the osmotic and heat stress. We have also observed that the osmotic stress induced important changes to the bacterial cell morphology. The production of the thermostable direct hemolysin, the major virulence factor in V. parahaemolyticus, was increased in cells remained viable after thermal inactivation at 47oC, these results demonstrating the risk of virulence conservation in insufficiently heat treated food.

Mechanism for decrease of glucocorticoid receptor mRNA in human hepatic cancer cell line SMMC-7721 during heat shock / 第二军医大学学报

Objective: To investigate the mechanism for decrease of glucocorticoid receptor mRNA (GR mRNA) during heat shock response. Methods: The changes of GR mRNA level in SMMC-7721 cells were examined by semi-quantitative RT-PCR during heat shock response. We designed 2 RNA fragments paired with GR gene intron E and cross exon 3, 4, and 5, which were used as probes for in situ hybridization with the sequences in SMMC-7721 cells at different periods after heat shock. The result was subjected to confocol microscope observation and computer image analysis. The transcription, degradation, and splicing of GR mRNA were investigated. Results: RT-PCR showed that the GR mRNA level was decreased during heat shock response. In situ hybridization revealed that both GR mRNA and GR pre-mRNA levels were lower in the heat shock group than in the non-heat shock group (P<0.05). GR pre-mRNA level was higher in cells treated with actinomycine D before heat shock than in cells only treated with actinomyeine D, while the GR mRNA level was lower in cells treated with actinomycine D before heat shock (P<0.05). Conclusion: During heat shock response, GR mRNA transcription is suppressed, the splicing of GR pre-mRNA is suppressed, and GR mRNA degradation is accelerated.

Cellular phenotype conversion and relative mechanism of human mesenchymal stem cells cocultured indirectly with human sweat gland cells: a preliminary study / 中华创伤杂志

Objective To observe the cellular phenotype conversion of human mesenchymal stem cells (MSCs) cocultured indirectly with heat-shocked human sweat gland cells (SGCs) in vitro and explore the relative mechanism. Methods MSCs and SGCs were isolated and amplified in vitro. First,primary confluent cultures of SGCs were heat-shocked at 47℃. Then, the supernatants were collected immediately and 24 hours before applied to the third generation of MSCs. After seven days, the MSCs expressing CK7, CK18 and CEA were examined by two-step immunocytochemistry and flow cytometry and compared with the control group. Results MSCs treated with the supernatants of SGCs proliferated slowly, with no obvious morphological changes during seven days. Two-step immunocytochemistry demonstrated positive staining of CK7 and CEA in some cells. Additionally, the positive rate of CK7 and CEA was 5.76% and 2.01% by flow cytometry, much higher than that of the control sample, which was only 1.12% and 0.51% respectively (P ＜ 0.01 ). Conclusions There are some signal moleculars in the supernatants of heat-shocked SGCs, which benefits the transdifferentiation of MSCs.

In vitro anti-tumor assay of dendritic cells sensitized by heat shock and OK-432 treated PANC1 lysate / 中华胰腺病杂志

treated with heat shock and OK-432 demonstrated enhanced biological activities,and could induce host lymphocytes to highly effective and specific eytotoxieity against PANC1 cells.

Repair potential of heat-treated rat bone marrow mesenchymal stem cells in infarcted myocardium / 中华器官移植杂志

Objective To study the effects of heat shock treatment of rat bone marrow mesenehymal stem cells(MSCs),the apoptosis ratio of treated-cells under low serum condition and the treated-cells transplantation on left ventricular function in rats with myocardiaIinfarction.Methods MSC8 were heat-treated under 42℃for 30 min,then the heat shock protein-70(HSP-70)was detected bv Western blot.The apoptosis ratio of heat-treated MSCs under low serum condition was tested by Annexin kit.The treated-MSCs labeled with Dil were transplanted into infarcted myocardium and 8 weeks later,the cardiac function of rats in each group was evaluated by echocardiography and cardiac catheterization.Results The immunophenotype of heat-treated MSCs did not vary,Western blot confirmed a higher level expression of HSP-70 in the treated-MSCs group as compared with that in the control group.The early apoptosis ratio was lower in treated-MSCs measured by flow cytometry with annexin staining than that of MSCs when cultured with low serum medium.After 8 weeks,LVEF,LVSP,+dp/dtmax,and-dp/dtmax were significantly higher,and the LVEDP was significantly lowar in heat-treated MSCs transplantation group than that in the control group.Conclusions Heat shock pretreatment of MSCs enhances the tolerance of MSCs to low serum medium,whereas does not lcad to the change of the cell immunophenotype.Transplantation of heattreated MSCs might improve the cardiac function in a rat myocardialinfarction model.

Hsp27 expression in osteoblast by thermal stress

Histopathologic evaluation of the peritoneum exposed to heat shock: experimental study in rats

PURPOSE: To evaluate histopathologic alterations of the peritoneum exposed to heat shock. METHODS: Sixty rats were randomly distributed into 6 groups: Heat Shock (HS), High Temperature (HT), Body Temperature (BT), Temperature 0oC (TZ), Sham (SH) and Control (CG) with 10 animals each. The peritoneal cavity of animals from groups HS, HT, BT and TZ was irrigated with NaCl solution 0.9 percent at temperatures 50°C, 0°C, 50°C, 37°C and 0°C, respectively. For animals from group SH, the procedures were simulated and those from group CG, laparotomy and biopsies were conducted. Twenty-four hours later, biopsies of the peritoneum for exams under light and electronic microscopy were performed. RESULTS: Edema was found in groups HS 80 percent, HT 60 percent, BT 30 percent TZ 70 percent, SH 40 percent and CG 30 percent. Vascular congestion was found in groups HS 20 percent, HT 30 percent, BT 10 percent and TZ 20 percent. Erythrocyte extravasation was found in groups HT 60 percent and SH 10 percent. Mesothelium destruction was found in 100 percent of specimens from groups HS, HT, BT, TZ, SH and CG 90 percent. Necrosis was found in groups HS 30 percent, HT 20 percent and BT 10 percent. The mean peritoneal thickness ranged from 42.26 μm (TZ) to 26.42 μm (CG). CONCLUSION: The heat shock caused no deaths, but promoted significant peritoneal edema without affecting the other histopathologic indicatives.

OBJETIVO: Avaliar alterações histopatológicas do peritônio exposto a choque térmico. MÉTODOS: Sessenta ratos foram distribuídos aleatoriamente em seis grupos: Choque Térmico (CT), Temperatura Elevada (TE), Temperatura 0°C (TZ) Sham (SH) e Controle (GC) com 10 animais. A cavidade peritoneal dos animais dos grupos CT, TE, TC e TZ foi irrigada com solução de NaCl 0,9 por cento nas temperaturas, 50°C e 0°C, 50°C, 37°C e 0°C, respectivamente. Nos animais do grupo SH foram simulados os procedimentos e nos do GC laparotomia e biópsias. Depois de 24 horas foram realizadas biópsias do peritônio para exames sob microscopia de luz e eletrônica. RESULTADOS: Edema foi encontrado nos grupos CT 80 por cento, TE 60 por cento, TC 30 por cento, TZ 70 por cento, SH 40 por cento e GC 30 por cento. Congestão vascular foi encontrada nos grupos CT 20 por cento, TE 30 por cento, TC 10 por cento e TZ 20 por cento. Extravasamento de hemácias foi encontrado nos grupos TE 60 por cento e SH 10 por cento. Destruição de mesotélio foi encontrada em 100 por cento dos espécimes dos grupos CT, TE, TC, TZ, SH e no grupo GC 90 por cento. Necrose foi encontrada nos grupos CT 30 por cento, TE 20 por cento e TC 10 por cento. A espessura média do peritônio variou de 42,26 μm (TZ) a 26,42 μm (GC). CONCLUSÃO: O choque térmico não causou óbitos, mas promoveu edema peritoneal significante sem alterar os demais indicadores histopatológicos.

Increase of NKG2D ligands and sensitivity to NK cell-mediated cytotoxicity of tumor cells by heat shock and ionizing radiation

In this study, we have investigated if current cancer therapeutic modalities including hyperthermia and ionizing radiation can increase the expression of NKG2D ligands in human cancer cell lines. The expressions of NKG2D ligands were induced by both heat shock and ionizing radiation in various cell lines including KM12, NCI-H23, HeLa and A375 cells with peaks at 2 h and 9 h after treatment, respectively, although inducibility of each NKG2D ligand was various depending on cell lines. During the induction of NKG2D ligands, heat shock protein 70 was induced by heat shock but not by ionizing radiation. These results were followed by increased susceptibilities to NK cell-mediated cytolysis after treatment with heat shock and ionizing radiation. These results suggest that heat shock and ionizing radiation induce NKG2D ligands and consequently might lead to increased NK cell-mediated cytotoxicity in various cancer cells.

Representational difference analysis of cDNA identifies novel genes expressed following preconditioning of the heart

Preconditioning of the myocardium rapidly induces a number of transcription factors, which are likely to be responsible for a cascade of transcriptional changes underlying the development of delayed adaptation. Identifying these changes provides insight into the molecular pathways elicited by sub-lethal ischaemia and the mechanism leading to delayed adaptation. Genes up-regulated in rabbit myocardium in vivo by ischaemic preconditioning following reperfusion for 2 h, 4 h and 6 h posttreatment were identified by representational difference analysis of cDNA (cDNA. RDA). The area of the left ventricle rendered ischaemic by preconditioning or the equivalent area of sham-treated animals was isolated and cDNA.RDA performed. Three novel genes and six genes with known function where identified, including the TGFbeta receptor interacting protein 1, the alpha isoform of the A subunit of PP2 and the cap binding protein NCBP1. To determine whether expression of these genes correlated with preconditioning per se, expression was measured in myocardium after both ischaemic as well as heat shock induced preconditioning following 2 h, 4 h, and 6 h reperfusion. These genes were induced in rabbit myocardium in vivo by both ischaemia and heat shock, consistent with a fundamental role in the development of delayed adaptation. The well described role of PP2 in modulating the mitogen-activated protein kinase pathway and promoting cell survival is consistent with our previous work, which identified the reperfusion injury salvage kinase pathway in mediating the protective effects of ischaemic preconditioning. Expression of Trip1 and Ncbp1 also implicates TGFbeta signalling pathways and RNA processing and transport in delayed adaptation to stress in the myocardium.

Quantitative analysis of the patterns of heat shock protein 70 expression induced by Hantaan virus infection in vitro / 中华传染病杂志

Objective To explore the pattern and quantify the heat shock protein HSP)70 and HSP70 mRNA in Vero-E6 cells after infection with Hantann virus(HTNV).Methods The expres- sion of HSP70 and change of its mRNA level were detected by immunocytochemical staining,nucleic acid hybridization in situ and RT-PCR.Results In situ hybridization and RT-PCR were used to eval- uate the level of HSP70 mRNA during Hantaan 76-118 infection.HSP70 mRNA increased 0.5 h after infection,reached its peak by 12 h and gradually declined to steady state level by 72 h(vs.sham infec- ted group,P＜0.05).The expression of HSP70 protein induced by Hantaan 76-118 infection was e- valuated by quantitative immunocytochemical staining.HSP70 increased 0.5 h after infection,reached its peak by 12 h and decreased at 72 h after infection(vs.sham infected group,P＜0.05).Conclu- sions HSP70 can be induced directly by HTNV infection at both mRNA and protein levels,It pro- vides a basis for the further study of the pathogenesis,prevention and treatment of hemorrhagic fever with renal syndrome(HFRS).

Protective effects and mechanism of heat shock response on cardiovascular system in rats after heat exposure / 中国病理生理杂志

AIM: To study protective effects and mechanism of heat shock response (HSR) on cardiovascular system in rats after heat exposure. METHODS: The study was divided into 2 experiments: ① Protective effects of HSR on cardiovascular system in rats after heat exposure. SD rats randomly allocated into 2 groups: heat shock group (HS group), sham control group (SC group). HS group were treated with heat shock, but SC group weren't. After recovering for 20 h at room temperature, two groups exposed to death in thermal environment, and blood pressure and electrocardiogram were measured continuously. Through Chart software mean arterial pressure(MAP), existent time etc were acquired. ② SD male rats randomly allocated into 3 groups: HS group, SC group and normal temperature control group (NC group). NC group weren't treated. The treatment in HS and SC group was identical with in the first experiment, but it would be terminated at 73 min after heat exposure, meanwhile content of MDA of myocardium were measured. RESULTS: ① Existent time in HS group was longer than that in SC group and shock arrived later; ② During earlier period after heat exposure MAP had no significant changes between HS and SC group, but after 60 mins MAP in HS group were higher than that in SC group; ③ Compared with NC group, content of MDA in myocardium in SC group was higher significantly at 73 min after heat exposure. Howerer, content of MDA in HS group was lower than in SC group, and had no significant changes with NC group. CONCLUSION:Through decreasing production of MDA in myocardium, HSR has a protective effect on cardiovascular system in rats after heat exposure.