The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation ...The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation of these genes remain largely unknown.Here,through comparative transcriptome and chromatin immunoprecipitation sequencing(ChIP-seq)analyses of the methyltransferase PoKmt6 in rice blast fungus Pyricularia oryzae(syn.Magnaporthe oryzae),we found that PoKmt6-mediated H3K27me3 deposition was enriched mainly at fast-evolving regions and contributed to the silencing of a subset of secreted proteins(SP)and transposable element(TE)families during the vegetative growth of P.oryzae.Intriguingly,we observed that a group of SP genes,which were depleted of H3K27me3 modification,could also be silenced via the H3K27me3-mediated repression of the nearby TEs.In conclusion,our results indicate that H3K27me3 modification mediated by PoKmt6 regulates the expression of some SP genes in fast-evolving regions through the suppression of nearby TEs.展开更多
Non-alcoholic fatty liver disease(NAFLD)is associated with mutations in lipopolysaccharide-binding protein(LBP),but the underlying epigenetic mechanisms remain understudied.Herein,LBP^(-/-)rats with NAFLD were establi...Non-alcoholic fatty liver disease(NAFLD)is associated with mutations in lipopolysaccharide-binding protein(LBP),but the underlying epigenetic mechanisms remain understudied.Herein,LBP^(-/-)rats with NAFLD were established and used to conduct integrative targetingactive enhancer histone H3 lysine 27 acetylation(H3K27ac)chromatin immunoprecipitation coupled with high-throughput and transcriptomic sequencing analysis to explore the potential epigenetic pathomechanisms of active enhancers of NAFLD exacerbation upon LBP deficiency.Notably,LBP^(-/-)reduced the inflammatory response but markedly aggravated high-fat diet(HFD)-induced NAFLD in rats,with pronounced alterations in the histone acetylome and regulatory transcriptome.In total,1128 differential enhancer-target genes significantly enriched in cholesterol and fatty acid metabolism were identified between wild-type(WT)and LBP^(-/-)NAFLD rats.Based on integrative analysis,CCAAT/enhancer-binding proteinβ(C/EBPβ)was identified as a pivotal transcription factor(TF)and contributor to dysregulated histone acetylome H3K27ac,and the lipid metabolism gene SCD was identified as a downstream effector exacerbating NAFLD.This study not only broadens our understanding of the essential role of LBP in the pathogenesis of NAFLD from an epigenetics perspective but also identifies key TF C/EBPβand functional gene SCD as potential regulators and therapeutic targets.展开更多
The effects of sodium salicylate on the expression of heat shock protein 27 （HSP27） during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells （HL...The effects of sodium salicylate on the expression of heat shock protein 27 （HSP27） during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells （HLB-3） were divided into 3 groups： control group （group A）, oxidation injury group （group B） and sodium salicylate group （group C）. Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium （MTT） chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system, Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B （0.2667±0.01414 vs 0.2150±0.01080, P=0.012〈0.05）. The average gray value of HSP27 in group B was less than that in group C （P=0.000〈0.05）. The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.展开更多
The heat shock proteins （HSPs） 70 and HSP 27 expression in patients with non-small cell lung cancer （NSCLC） was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCL...The heat shock proteins （HSPs） 70 and HSP 27 expression in patients with non-small cell lung cancer （NSCLC） was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients＇ clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 （78.3 %） and 43 （71.7 %） specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differentiation, lymph node metastasis, patients＇ clinical stages, smoking history, whereas HSP 27 expression is not.展开更多
The herpes simplex virus type 1 （HSV-1） infected-cell protein 27 （ICP27） is an essential, highly conserved protein involved in various steps of HSV-1 gene regulation as well as in the shut-off of host gene express...The herpes simplex virus type 1 （HSV-1） infected-cell protein 27 （ICP27） is an essential, highly conserved protein involved in various steps of HSV-1 gene regulation as well as in the shut-off of host gene expression during infection. It functions primarily at the post-transcriptional level in inhibiting precursor mRNA splicing and in promoting nuclear export of viral transcripts. Recently, many novel functions performed by the HSV- 1 ICP27 protein were shown, including leptomycin B resistance, inhibition of the type I interferon signaling, regulation of the viral mRNA translation and determining the composition of HSV-1 virions展开更多
Objective： To clone human cardiac heat shock protein 27 （HSP27） gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods： Full length of HSP27 cDNA which got ...Objective： To clone human cardiac heat shock protein 27 （HSP27） gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods： Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1^＋ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable trahsfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H2O2 in H9c2. Results： ①pCDNA3.1^＋/HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H2O2 in HSP27 over-expression group and wild type group were 0.396±0.017 vs. 0.390±0.01）9 （p 〉0.05）, 0.437±0. 014 vs. 0.416±0.015 （P〈0.05）, 0.471±0.018 vs. 0.417±0.009 （P 〈0.001）, 0.505±0.030 vs. 0.657± 0.022（P 〈0.001）, 0.547 ±0.027 and 0.661 ± 0.011（ P 〈 0. 001 ）, respectively. ③Apoptosis induced by 150 μmol/L H2O2 in HSP27 over-expression group and wild type group were （10.693± 1.122）% vs. （4.027 ± 1.628）%（ P 〈0.01）. Conclusion： We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.展开更多
To study the expression of heat stress protein 27 (HSP27) and P21 in nasopharyngeal car- cinoma (NPC) tissue, and to evaluate the significance of both HSP27 and P21 in the pathogenesis, development and prognosis of N...To study the expression of heat stress protein 27 (HSP27) and P21 in nasopharyngeal car- cinoma (NPC) tissue, and to evaluate the significance of both HSP27 and P21 in the pathogenesis, development and prognosis of NPC. Indirect immunofluorescence method combined with SABC was applied. Our results showed that (1) the positive rates of HSP27 and P21 expressed in NPC tissue in 36 cases were 88. 9 % and 94. 4%; (2) while in 10 hyperplastic nasopharyngitis tissues, the positive rate of HSP27 and P21 were both 5; (3) all the 5 normal tissues were negatively stained. It is obvi- ous that a co-expressing tendency of HSP27 and P21 could be identified, and it was associated with the degree of malignancy and the clinical stage of NPC. It is concluded that the positive expression of HSP27 and P21 may have clinical significance in the evaluation of the occurring, development and prognosis of NPC, and in NPC treatment.展开更多
OBJECTIVE: To investigate the effect of inhibiting factor of cell cycle regulation p27<sup>kipl</sup>,retinoblastinoma protein （Rb protein）, and proliferating cell nuclear antigen （PCNA） on the genesis...OBJECTIVE: To investigate the effect of inhibiting factor of cell cycle regulation p27<sup>kipl</sup>,retinoblastinoma protein （Rb protein）, and proliferating cell nuclear antigen （PCNA） on the genesis andprogression of human pancreatic cancer.METHODS: The expression of p27<sup>kipl</sup>, Rb protein and PCNA in the tumor tissue and adjacent tissue of32 patients with pancreatic cancer was detected by SP immunohistochemical technique.RESULTS: The p27<sup>kipl</sup> protein positive-expression rate in the tumor tissue of pancreatic cancer was56.25%, which was lower than that in the adjacent pancreatic tissue （P【0.05）. p27<sup>kipl</sup> proteinpositive-expression was correlated significantly with tumor cell differentiation and lymph node metastasis（P【0.05）. The Rb gene protein positive-expression rate in the tumor tissue was 50%, which was alsolower than that in the adjacent pancreatic tissue （P【0.05 ）. The PCNA positive-expression rate was71.87%, which was higher than that in the adjacent pancreatic tissue （P【0.05）. PCNA positive-expression was also correlated significantly with tumor cell differentiation and lymph node metastasis（P【0.05）.CONCLUSION: The decreased expression of p27<sup>kipl</sup>, Rb protein and over-expression of PCNA may playan important role in the genesis and progression of pancreatic cancer.展开更多
AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glauc...AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glaucoma model and disclose the underlying pathogenesis of glaucoma.METHODS: A total of 115 Wistar rats were randomly divided into 4 groups. Group 1 was the ocular hypertension group by condensing 3 episcleral & limbal veins or episcleral area of right eye(HP group, n=25) and sham operation group with conjunctiva incision without coagulation(n=25). Group 2: HSP27 or dose-matched PBS was injected into the vitreous(V-HSP27 group, n=15;V-PBS group, n=15). Group 3: HSP27 and complete Freund's adjuvant or dosematched PBS was injected subcutaneously into the hind limb accompanied intraperitoneal injection of pertussis toxin [sensitized group(I-HSP27 group), n=15;I-PBS group, n=15)]. Group 4 was normal group without any treatment(n=5). IOPs of the rats were measured before, day 3, weeks 1, 2, 4, 6, and 8 after treatment. Paraffin-embedded sections were prepared for HE staining and RGCs apoptosis were detected by TUNEL. Anti-HSP27 level in serum and CSF were examined by ELISA. RESULTS: IOPs were elevated significantly in HP and V-HSP27, V-PBS groups(P<0.01) and positively related to anti-HSP27 levels in serum and CSFs. Anti-HSP27 levels in serum and CSF were elevated significantly in I-HSP27group compared to other groups(P<0.05). However, the IOPs did not show any relationship with the high-level antiHSP27 in serum and CSFs. RGC apoptosis were all elevated significantly in the HP, V-HSP27, V-PBS and I-HSP27 groups and also positively relative with anti-HSP27 level in serum and CSFs except that high-level of anti-HSP27 in the serum of I-HSP group.CONCLUSION: The increases of anti-HSP27 levels in serum and CSFs both promote IOP escalation and the increase of RGC apoptosis in retina when anti-HSP27 is at low level. The case of high-level anti-HSP27 is opposite and shows protective function in preventing IOP increase and RGC apoptosis.展开更多
The p27Kip1 is a cell cycle repressor protein that regulates primarily the cell cycle transition from G1 to S phase and hence the DNA replication is in the S phase and cell division in the M phase. Expression of p27Ki...The p27Kip1 is a cell cycle repressor protein that regulates primarily the cell cycle transition from G1 to S phase and hence the DNA replication is in the S phase and cell division in the M phase. Expression of p27Kip1 protein has dual roles for both cancer prevention and promotion. For example, numerous nutritional and chemopreventive anti-cancer agents specifically increase the expression of p27Kip1 protein without directly affecting the expression of any other cell cycle regulatory proteins. On the other hand, pro-cancer agents (like glucose, insulin and other growth factors frequently seen in obesity and/or diabetes) specifically decrease the expression of p27Kip1 protein without directly affecting the expression of any other cell cycle regulatory proteins. Unlike expression of any other cell cycle regulatory proteins, expression of p27Kip1 protein is very unusual. The mRNA of p27Kip1 has a very long and unusual 5’-untranslated region (from -575 to -1 in human). It appears that the 5’-untranslated region of p27Kip1 mRNA forms two alternative secondary structures. One increases the expression of p27Kip1 protein when anti-cancer agents are added and another decrease the expression of p27K1p1 when pro-cancer agents are added. For this short concept proposal, Dr. Albert Einstein’s “visualized thought experiments (German: Gedanken experiment)” were used as a fundamental tool for understanding how either anti- or pro-cancer agents bring the primary structure of the 5’-untranslated region of p27Kip1 mRNA into two alternative secondary structures, thereby either increasing or decreasing, respectively, the translation initiation of p27Kip1 protein.展开更多
Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were f...Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h.Thereafter,the protein expression of HSP-27 and HSP-70 was evaluated by western blotting.Alternatively,HZ-fed cells were cultured up to 72 h and cell viability parameters(survival,apoptosis and necrosis rates) were measured by flow cytometric analysis. Results:HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells,and promoted long-term cell survival without inducing apoptosis.As expected,gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis.Conclusions: Present data show that HZ prevents cell apoptosis and enhances the expression of anli-apoptotic HSP-27 in THP-1 cells,confirming the previous evidences obtained from HZ-fed immunopurified monocytes.Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach,which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.展开更多
Quality protein maize(QPM)(Zea mays L.) varieties contain enhanced levels of tryptophan and lysine, exhibiting improved nutritive value for humans and livestock. However, breeding QPM varieties remains challenging due...Quality protein maize(QPM)(Zea mays L.) varieties contain enhanced levels of tryptophan and lysine, exhibiting improved nutritive value for humans and livestock. However, breeding QPM varieties remains challenging due to the complex process of rebalancing storage protein. This study conducted transcriptome and proteome analyses to investigate the process of storage proteins rebalancing in opaque2(o2) and QPM. We found a weak correlation between the transcriptome and proteome, suggesting a significant modulating effect of post-transcriptional events on non-zein protein abundances in Mo17o2 and QPM. These results highlight the advantages of proteomics. Compared with Mo17, 672 differentially expressed proteins(DEPs) were identified both in Mo17o2 and QPM, and several of them were associated with storage protein, starch, and amino acid synthesis. We identified 178 non-zeins as DEPs in Mo17o2 and QPM kernels. The up-regulated non-zein DEPs were enriched in lysine, tryptophan, and methionine, which affected the protein quality. Co-expression network analysis identified regulators of storage protein synthesis in QPM, including O2,PBF1, and several transcription factors. Our results revealed how storage protein rebalancing occurs and identified nonzein DEPs that may facilitate superior-quality QPM breeding.展开更多
Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene produ...Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni 2+ -NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.展开更多
基金the grants from the National Natural Science Foundation of China(U1805232,31770156,and 32172365)the China Postdoctoral Science Foundation(2021M690637)。
文摘The fine-tuned expression dynamics of the effector genes are pivotal for the transition from vegetative growth to host colonization of pathogenic filamentous fungi.However,mechanisms underlying the dynamic regulation of these genes remain largely unknown.Here,through comparative transcriptome and chromatin immunoprecipitation sequencing(ChIP-seq)analyses of the methyltransferase PoKmt6 in rice blast fungus Pyricularia oryzae(syn.Magnaporthe oryzae),we found that PoKmt6-mediated H3K27me3 deposition was enriched mainly at fast-evolving regions and contributed to the silencing of a subset of secreted proteins(SP)and transposable element(TE)families during the vegetative growth of P.oryzae.Intriguingly,we observed that a group of SP genes,which were depleted of H3K27me3 modification,could also be silenced via the H3K27me3-mediated repression of the nearby TEs.In conclusion,our results indicate that H3K27me3 modification mediated by PoKmt6 regulates the expression of some SP genes in fast-evolving regions through the suppression of nearby TEs.
基金supported by the National Natural Science Foundation of China(81971875,82300661)Natural Science Foundation of Anhui province(2308085QH246)+3 种基金Natural Science Foundation of the Anhui Higher Education Institutions(KJ2021A0205)Basic and Clinical Cooperative Research Program of Anhui Medical University(2019xkjT002,2019xkjT022,2022xkjT013)Talent Training Program,School of Basic Medical Sciences,Anhui Medical University(2022YPJH102)National College Students Innovation and Entrepreneurship Training Program of China(202210366024)。
文摘Non-alcoholic fatty liver disease(NAFLD)is associated with mutations in lipopolysaccharide-binding protein(LBP),but the underlying epigenetic mechanisms remain understudied.Herein,LBP^(-/-)rats with NAFLD were established and used to conduct integrative targetingactive enhancer histone H3 lysine 27 acetylation(H3K27ac)chromatin immunoprecipitation coupled with high-throughput and transcriptomic sequencing analysis to explore the potential epigenetic pathomechanisms of active enhancers of NAFLD exacerbation upon LBP deficiency.Notably,LBP^(-/-)reduced the inflammatory response but markedly aggravated high-fat diet(HFD)-induced NAFLD in rats,with pronounced alterations in the histone acetylome and regulatory transcriptome.In total,1128 differential enhancer-target genes significantly enriched in cholesterol and fatty acid metabolism were identified between wild-type(WT)and LBP^(-/-)NAFLD rats.Based on integrative analysis,CCAAT/enhancer-binding proteinβ(C/EBPβ)was identified as a pivotal transcription factor(TF)and contributor to dysregulated histone acetylome H3K27ac,and the lipid metabolism gene SCD was identified as a downstream effector exacerbating NAFLD.This study not only broadens our understanding of the essential role of LBP in the pathogenesis of NAFLD from an epigenetics perspective but also identifies key TF C/EBPβand functional gene SCD as potential regulators and therapeutic targets.
文摘目的分析老年急性脑梗死(acute cerebral infarction,ACI)患者血清氧化低密度脂蛋白(oxidized low density lipoprotein,OX-LDL)、热休克蛋白27(heat shock proteins 27,HSP27)水平与脑水肿严重程度及短期预后的关系。方法回顾性分析2019年1月—2021年9月郑州大学第五附属医院收治的初发老年ACI患者120例(研究组)住院后当天,第3、5、7天的病例资料,按照患者住院当天美国国立卫生院卒中量表(national institute health stroke scale,NIHSS)评分分为轻度组(n=36)、中度组(n=50)、重度组(n=34),于发病后90天,根据改良Rankin量表(modified Rankin Scale,mRS)评分分为预后良好组(n=82)和预后不良组(n=38);另取同期门诊健康体检者120例为对照组。采用酶联免疫吸附试验(ELISA)检测血清OX-LDL、HSP27水平,采用BORN-BE无创脑水肿动态监护仪检测患侧及对照组同侧大脑半球脑电阻抗扰动系数(coefficient of electrical impedance,CEI),采用Pearson法分析血清OX-LDL、HSP27水平与CEI扰动系数的相关性。结果与对照组比较,轻度组、中度组、重度组ACI患者NIHSS评分依次升高,差异有统计学意义(P<0.05)。与对照组比较,轻度组、中度组、重度组ACI患者血清OX-LDL水平、CEI依次增加,差异有统计学意义(P<0.05),均呈先增加后降低趋势,在第5天达到高峰,于第7天开始下降;血清HSP27水平依次降低,差异有统计学意义(P<0.05),随时间变化趋势与OX-LDL相反。Pearson相关性分析结果显示,ACI患者各时间点血清OX-LDL与CEI均呈正相关(P<0.05),血清HSP27水平与CEI均负相关(P<0.05)。与预后良好组比较,预后不良组患者各时间点血清OX-LDL水平升高,差异有统计学意义(P<0.05),随住院时间延长呈先升高后降低趋势,且在第5天达到高峰,第7天开始下降;血清HSP27水平降低,差异有统计学意义(P<0.05),随时间变化趋势与OXLDL相反。结论血清OX-LDL、HSP27水平变化与ACI患者病情、脑水肿严重程度及预后有关,可能对ACI患者脑水肿严重程度及预后评估有一定临床指导意义。
文摘The effects of sodium salicylate on the expression of heat shock protein 27 （HSP27） during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells （HLB-3） were divided into 3 groups： control group （group A）, oxidation injury group （group B） and sodium salicylate group （group C）. Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium （MTT） chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system, Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B （0.2667±0.01414 vs 0.2150±0.01080, P=0.012〈0.05）. The average gray value of HSP27 in group B was less than that in group C （P=0.000〈0.05）. The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.
文摘The heat shock proteins （HSPs） 70 and HSP 27 expression in patients with non-small cell lung cancer （NSCLC） was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients＇ clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 （78.3 %） and 43 （71.7 %） specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differentiation, lymph node metastasis, patients＇ clinical stages, smoking history, whereas HSP 27 expression is not.
基金Start Fund of the Hundred Talents Program of the Chinese Academy of Science (20071010-141)National Natural Science Foundation of China(30870120)Open Research Fund Program of the State Key Laboratory of Virology of China (2007003)
文摘The herpes simplex virus type 1 （HSV-1） infected-cell protein 27 （ICP27） is an essential, highly conserved protein involved in various steps of HSV-1 gene regulation as well as in the shut-off of host gene expression during infection. It functions primarily at the post-transcriptional level in inhibiting precursor mRNA splicing and in promoting nuclear export of viral transcripts. Recently, many novel functions performed by the HSV- 1 ICP27 protein were shown, including leptomycin B resistance, inhibition of the type I interferon signaling, regulation of the viral mRNA translation and determining the composition of HSV-1 virions
文摘Objective： To clone human cardiac heat shock protein 27 （HSP27） gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods： Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1^＋ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable trahsfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H2O2 in H9c2. Results： ①pCDNA3.1^＋/HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H2O2 in HSP27 over-expression group and wild type group were 0.396±0.017 vs. 0.390±0.01）9 （p 〉0.05）, 0.437±0. 014 vs. 0.416±0.015 （P〈0.05）, 0.471±0.018 vs. 0.417±0.009 （P 〈0.001）, 0.505±0.030 vs. 0.657± 0.022（P 〈0.001）, 0.547 ±0.027 and 0.661 ± 0.011（ P 〈 0. 001 ）, respectively. ③Apoptosis induced by 150 μmol/L H2O2 in HSP27 over-expression group and wild type group were （10.693± 1.122）% vs. （4.027 ± 1.628）%（ P 〈0.01）. Conclusion： We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.
文摘To study the expression of heat stress protein 27 (HSP27) and P21 in nasopharyngeal car- cinoma (NPC) tissue, and to evaluate the significance of both HSP27 and P21 in the pathogenesis, development and prognosis of NPC. Indirect immunofluorescence method combined with SABC was applied. Our results showed that (1) the positive rates of HSP27 and P21 expressed in NPC tissue in 36 cases were 88. 9 % and 94. 4%; (2) while in 10 hyperplastic nasopharyngitis tissues, the positive rate of HSP27 and P21 were both 5; (3) all the 5 normal tissues were negatively stained. It is obvi- ous that a co-expressing tendency of HSP27 and P21 could be identified, and it was associated with the degree of malignancy and the clinical stage of NPC. It is concluded that the positive expression of HSP27 and P21 may have clinical significance in the evaluation of the occurring, development and prognosis of NPC, and in NPC treatment.
文摘OBJECTIVE: To investigate the effect of inhibiting factor of cell cycle regulation p27<sup>kipl</sup>,retinoblastinoma protein （Rb protein）, and proliferating cell nuclear antigen （PCNA） on the genesis andprogression of human pancreatic cancer.METHODS: The expression of p27<sup>kipl</sup>, Rb protein and PCNA in the tumor tissue and adjacent tissue of32 patients with pancreatic cancer was detected by SP immunohistochemical technique.RESULTS: The p27<sup>kipl</sup> protein positive-expression rate in the tumor tissue of pancreatic cancer was56.25%, which was lower than that in the adjacent pancreatic tissue （P【0.05）. p27<sup>kipl</sup> proteinpositive-expression was correlated significantly with tumor cell differentiation and lymph node metastasis（P【0.05）. The Rb gene protein positive-expression rate in the tumor tissue was 50%, which was alsolower than that in the adjacent pancreatic tissue （P【0.05 ）. The PCNA positive-expression rate was71.87%, which was higher than that in the adjacent pancreatic tissue （P【0.05）. PCNA positive-expression was also correlated significantly with tumor cell differentiation and lymph node metastasis（P【0.05）.CONCLUSION: The decreased expression of p27<sup>kipl</sup>, Rb protein and over-expression of PCNA may playan important role in the genesis and progression of pancreatic cancer.
基金Supported by National Natural Science Foundation of China(No.81060077)Key Research and Development Program in Yunnan Province(International Science and Technology Cooperation,No.2017IB001)Science and Technology Planning Project of Guangzhou(No.201604020105)。
文摘AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glaucoma model and disclose the underlying pathogenesis of glaucoma.METHODS: A total of 115 Wistar rats were randomly divided into 4 groups. Group 1 was the ocular hypertension group by condensing 3 episcleral & limbal veins or episcleral area of right eye(HP group, n=25) and sham operation group with conjunctiva incision without coagulation(n=25). Group 2: HSP27 or dose-matched PBS was injected into the vitreous(V-HSP27 group, n=15;V-PBS group, n=15). Group 3: HSP27 and complete Freund's adjuvant or dosematched PBS was injected subcutaneously into the hind limb accompanied intraperitoneal injection of pertussis toxin [sensitized group(I-HSP27 group), n=15;I-PBS group, n=15)]. Group 4 was normal group without any treatment(n=5). IOPs of the rats were measured before, day 3, weeks 1, 2, 4, 6, and 8 after treatment. Paraffin-embedded sections were prepared for HE staining and RGCs apoptosis were detected by TUNEL. Anti-HSP27 level in serum and CSF were examined by ELISA. RESULTS: IOPs were elevated significantly in HP and V-HSP27, V-PBS groups(P<0.01) and positively related to anti-HSP27 levels in serum and CSFs. Anti-HSP27 levels in serum and CSF were elevated significantly in I-HSP27group compared to other groups(P<0.05). However, the IOPs did not show any relationship with the high-level antiHSP27 in serum and CSFs. RGC apoptosis were all elevated significantly in the HP, V-HSP27, V-PBS and I-HSP27 groups and also positively relative with anti-HSP27 level in serum and CSFs except that high-level of anti-HSP27 in the serum of I-HSP group.CONCLUSION: The increases of anti-HSP27 levels in serum and CSFs both promote IOP escalation and the increase of RGC apoptosis in retina when anti-HSP27 is at low level. The case of high-level anti-HSP27 is opposite and shows protective function in preventing IOP increase and RGC apoptosis.
文摘The p27Kip1 is a cell cycle repressor protein that regulates primarily the cell cycle transition from G1 to S phase and hence the DNA replication is in the S phase and cell division in the M phase. Expression of p27Kip1 protein has dual roles for both cancer prevention and promotion. For example, numerous nutritional and chemopreventive anti-cancer agents specifically increase the expression of p27Kip1 protein without directly affecting the expression of any other cell cycle regulatory proteins. On the other hand, pro-cancer agents (like glucose, insulin and other growth factors frequently seen in obesity and/or diabetes) specifically decrease the expression of p27Kip1 protein without directly affecting the expression of any other cell cycle regulatory proteins. Unlike expression of any other cell cycle regulatory proteins, expression of p27Kip1 protein is very unusual. The mRNA of p27Kip1 has a very long and unusual 5’-untranslated region (from -575 to -1 in human). It appears that the 5’-untranslated region of p27Kip1 mRNA forms two alternative secondary structures. One increases the expression of p27Kip1 protein when anti-cancer agents are added and another decrease the expression of p27K1p1 when pro-cancer agents are added. For this short concept proposal, Dr. Albert Einstein’s “visualized thought experiments (German: Gedanken experiment)” were used as a fundamental tool for understanding how either anti- or pro-cancer agents bring the primary structure of the 5’-untranslated region of p27Kip1 mRNA into two alternative secondary structures, thereby either increasing or decreasing, respectively, the translation initiation of p27Kip1 protein.
基金supported by University of Torino Intramural Funds to GG and by grants to MP from the Compagnia di San Paolo,Torino,in the context of the Italian Malaria Network
文摘Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h.Thereafter,the protein expression of HSP-27 and HSP-70 was evaluated by western blotting.Alternatively,HZ-fed cells were cultured up to 72 h and cell viability parameters(survival,apoptosis and necrosis rates) were measured by flow cytometric analysis. Results:HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells,and promoted long-term cell survival without inducing apoptosis.As expected,gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis.Conclusions: Present data show that HZ prevents cell apoptosis and enhances the expression of anli-apoptotic HSP-27 in THP-1 cells,confirming the previous evidences obtained from HZ-fed immunopurified monocytes.Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach,which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.
基金supported by the National Natural Science Foundation of China (31971951 and 31771796)。
文摘Quality protein maize(QPM)(Zea mays L.) varieties contain enhanced levels of tryptophan and lysine, exhibiting improved nutritive value for humans and livestock. However, breeding QPM varieties remains challenging due to the complex process of rebalancing storage protein. This study conducted transcriptome and proteome analyses to investigate the process of storage proteins rebalancing in opaque2(o2) and QPM. We found a weak correlation between the transcriptome and proteome, suggesting a significant modulating effect of post-transcriptional events on non-zein protein abundances in Mo17o2 and QPM. These results highlight the advantages of proteomics. Compared with Mo17, 672 differentially expressed proteins(DEPs) were identified both in Mo17o2 and QPM, and several of them were associated with storage protein, starch, and amino acid synthesis. We identified 178 non-zeins as DEPs in Mo17o2 and QPM kernels. The up-regulated non-zein DEPs were enriched in lysine, tryptophan, and methionine, which affected the protein quality. Co-expression network analysis identified regulators of storage protein synthesis in QPM, including O2,PBF1, and several transcription factors. Our results revealed how storage protein rebalancing occurs and identified nonzein DEPs that may facilitate superior-quality QPM breeding.
基金The startup fund of the hundred talents program of the Chinese academy of science(20071010- 141)National natural science foundation of China (30870120)+2 种基金Open research fund program of the state key laboratory of virology of China (2007003, 2009007)Hubei province natural science foundation of innovation groups project (2008CDA013)Major state basic research development program (973 Program) of China (2010CB 530105)
文摘Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni 2+ -NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.
