Dynamics and regulation of glycolysis‐tricarboxylic acid metabolism in the midgut of Spodoptera litura during metamorphosis

Significant changes usually take place in the internal metabolism of insects during metamorphosis. The glycolysis‐tricarboxylic acid (glycolysis‐TCA) pathway is important for energy metabolism. To elucidate its dynamics, the mRNA levels of genes involved in this pathway were examined in the midgut of Spodoptera litura during metamorphosis, and the pyruvate content was quantified. The expression patterns of these genes in response to starvation were examined, and the interaction between protein phosphatase 1 (PP1) and phosphofructokinase (PFK) was studied. The results revealed that the expression or activities of most glycolytic enzymes was down‐regulated in prepupae and then recovered in some degree in pupae, and all TCA‐related genes were remarkably suppressed in both the prepupae and pupae. Pyruvate was enriched in the pupal midgut. Taken together, these results suggest that insects decrease both glycolysis and TCA in prepupae to save energy and then up‐regulate glycolysis but down‐regulate TCA in pupae to increase the supply of intermediates for construction of new organs. The expression of all these genes were down‐regulated by starvation, indicating that non‐feeding during metamorphosis may be a regulator of glycolysis‐TCA pathway in the midgut. Importantly, interaction between PP1 and PFK was identified and is suggested to be involved in the regulation of glycolysis.     

The diagnostic value of urine heat shock protein 70 and prostatic exosomal protein in chronic prostatitis

ObjectiveTo explore the diagnostic value of the levels of prostatic exosomal protein (PSEP) and heat shock protein 70 (HSP70) in the urine of patients with chronic prostatitis (CP).MethodUrine samples from 210 CP patients (70 cases of the USA National Institutes of Health Category II [NIH‐II], 70 NIH‐IIIa, and 70 NIH‐IIIb patients) and 70 control subjects were collected between May 2018 and February 2020. The levels of PSEP and HSP70 in urine were detected by enzyme‐linked immunosorbent assay. The differences in urine PSEP and HSP70 levels between the groups were analyzed, and receiver operating characteristic (ROC) curves were used to analyze the clinical value of PSEP and HSP70 in the diagnosis of CP.ResultsThe PSEP levels of CP patients were significantly higher than those of the control group (p < 0.001), but there was no difference in PSEP levels among CP subgroups. The level of HSP70 in the urine of the NIH‐II patients was significantly lower than the levels in the NIH‐IIIa and NIH‐IIIb subgroups and the control group, but there was no difference in HSP70 levels between the NIH‐IIIa and NIH‐IIIb subgroups and the control group. ROC curve analysis results showed that the area under the curve (AUC) of PSEP for the NIH‐II, NIH‐IIIa, and NIH‐IIIb patients was 0.751, 0.776, and 0.731, respectively. The AUC of HSP70 in NIH‐II patients was 0.784, and the AUC of combined detection of PSEP and HSP70 in NIH‐II patients was 0.858.ConclusionUrine PSEP can be used as a marker for the diagnosis of CP, but it cannot distinguish between the various types of CP, and HSP70 can be used as a diagnostic index for NIH‐II classification.     

糖皮质激素受体和热休克蛋白70在大鼠创伤失血性休克肝组织中的变化及其意义

目的从组织细胞受体水平探讨糖皮质激素受体（GR）和热休克蛋白70（HSP70）在创伤失血性休克后肝脏中的变化及其作用。方法采用双侧股骨骨折伴失血性休克致严重创伤模型，动态观察伤后8h大鼠肝组织GR、HSP70、血清肝功能生化指标、肝脏病理等变化。采用蛋白质免疫印迹法（Western blot）测定肝组织GR、HSP70含量，并进行计算机图像分析。结果单纯股骨骨折伤后，肝组织GR含量于伤后1h即开始下降，6h降至最低，8h仍显著低于正常对照组；创伤合并休克后，GR下降更加明显。HSP70在单纯股骨骨折伤后迅速增加，6h达到峰值，8h仍持续在较高水平；创伤合并休克后，HSP70升高更加显著。单纯股骨骨折伤后，血清丙氨酸转氨酶（ALT）、总胆红素（TB）和白蛋白与正常对照组比较，差异均无显著性。但创伤合并休克后4h和2h血清ALT和TB分别开始明显增高（P均d〈0．01），白蛋白则均下降（P均〈0．01）。创伤合并休克后6h肝脏镜下肝窦内即出现较多炎性细胞浸润。结论GR不足在严重创伤失血性休克后肝脏继发性损害过程中起重要作用，HSP70可能参与了肝组织细胞抗损伤机制的启动。     

缺血后处理对大鼠缺血/再灌注心肌热休克蛋白的影响

目的:研究缺血后处理对大鼠缺血/再灌注心肌热休克蛋白(HSP70)的影响。方法:选择健康SD大鼠48只,随机分为3组：假手术组、缺血再灌注组（对照组）和缺血后处理组,每组16只。制备大鼠心肌缺血/再灌注模型。缺血再灌注组,收紧结扎线缺血40 min, 放松结扎线再灌注240min;缺血后处理组,缺血40 min后, 再灌注10s,缺血10s,连续3个循环,然后再灌注240min;假手术组,开胸后穿线做套环,但不收紧结扎线。免疫组织化学染色检测HSP70的表达,TUNEL 法检测心肌细胞凋亡指数,同时测定血清肌酸激酶活性。结果:①血清肌酸激酶活性测定：再灌注结束后缺血后处理组和缺血再灌注组肌酸激酶活性明显高于假手术组,分别为（712.13±42.77）,（935.17±57.99）,（282.74±29.54）U/L,P＜0.05,缺血后处理组明显低于对照组(P＜0.05)。②心肌凋亡细胞计数：再灌注结束后假手术组未见明显细胞凋亡（＜5%）,缺血后处理组心肌细胞凋亡率明显低于缺血再灌注组,分别为（14.3±2.7）%,（22.3±3.6）%,（P＜0.05）。③心肌热休克蛋白表达：缺血后处理组较对照组心肌热休克蛋白表达增强（P＜0.05）。结论:缺血后处理可减轻缺血再灌注损伤,其机制可能与增强热休克蛋白表达,减少心肌细胞凋亡有关。     

严重烧伤早期肠黏膜组织热休克蛋白70的表达规律

目的探讨大鼠烧伤后早期肠黏膜组织热休克蛋白70(HSP70)的表达变化规律及其意义。方法采用大鼠烫伤模型,通过逆转录-聚合酶链反应(RT-PCR)、蛋白质免疫印迹(Westernblot)及免疫组化等方法,检测伤后3、6、12、24和48h不同时间点肠黏膜组织内HSP70及热休克因子1(HSF1)的表达分布情况。结果烫伤后3h肠黏膜组织内HSP70mRNA及蛋白表达均显著增加,分别在伤后6h和12h达高峰,伤后48h仍高于正常对照组(P均<0.01);伤后3h大鼠肠黏膜组织HSF1出现一过性降低,伤后6h其表达显著高于正常对照组,并呈逐渐增加的趋势直至持续到伤后48h(P均<0.01)。结论严重烧伤早期肠黏膜组织HSP70及HSF1表达均显著增加,提示严重烧伤早期即可引起肠黏膜组织细胞的应激反应,可能与细胞的自我保护机制启动有关。     

热休克蛋白70与慢性阻塞性肺疾病的关系

目的探讨热休克蛋白70(HSP70)与慢性阻塞性肺疾病(COPD)的关系。方法选取COPD患者130例,按照暴露情况分为吸烟COPD组100例(按肺功能分为轻、中、重度)、不吸烟COPD组30例;同时选取对照组60例,按暴露情况分为吸烟对照组30例、不吸烟对照组30例。采用Westernblot法检测各组HSP70表达。分析四组及不同程度的吸烟COPD患者与HSP70的关系。结果吸烟COPD组、不吸烟COPD组、吸烟对照组和不吸烟对照组的HSP70吸光度分别为:19.22±5.33、21.96±7.85、22.65±10.01和22.21±8.94,吸烟COPD组中外周血HSP70的表达明显低于不吸烟COPD组、吸烟对照组及不吸烟对照组,差异均有统计学意义(t分别=2.41、3.25、2.98,P均<0.05);重、中、轻度COPD组的HSP吸光度分别为:17.92±5.81、24.13±13.11和26.82±14.93,重度的COPD患者外周血HSP70的表达明显低于中度及轻度的COPD患者,中度COPD患者低于轻度的COPD患者,差异均有统计学意义(t分别=2.50、3.93、3.18,P均<0.05)。线性回归分析显示:HSP70与COPD的严重程度呈负相关(r=-0.57,P<0.05)。结论 HSP70与COPD的发生相关,且HSP70表达的高低与COPD疾病的严重程度呈负相关。     

高血压脑梗死与血浆热休克蛋白70抗体水平关系的研究

目的通过测定高血压脑梗死患者血浆热休克蛋白70（HSP70）抗体水平，探讨HSP70抗体与脑梗死损伤程度的关系及其在脑梗死发病机制中的作用。方法100例脑梗死患者经头颅CT或MRI确诊，其中男64例，女36例；高血压者67例，血压正常者33例。选100名社区居民作为对照组，其中男58名，女42名；高血压者35名，血压正常者65名。采用蛋白质免疫印迹-酶联免疫吸附法（Western blot—ELISA）测定血浆HSP70抗体，并进行Logistic回归分析。结果脑梗死组HSP70抗体阳性率明显高于对照组（69％比13％）；脑梗死组高血压者HSP70抗体阳性率明显高于血压正常者（57％比12％），对照组高血压者HSP70抗体阳性率亦高于血压正常者（11％比2％）。上述结果差异均有显著性（P均〈0．05）。结论HSP70抗体增多与高血压脑梗死发病相关，可作为高血压脑缺血时一种分子生物学诊断指标；HSP70抗体表达越强，脑梗死病情越严重。因此，HSP70抗体增高是反映脑组织损伤程度的标志。     

Induction of heat shock protein 70 genes in human lymphocytes during fever therapy

The induction of heat shock protein 70 ( HSP70 ) genes has been studied in peripheral blood mononuclear cells of individuals undergoing fever therapy because of metastatic malignant melanoma. Induction of HSP70 was assessed at the protein level by metabolic labelling or, for the HLA-linked HSP70-1 and HSP70-2 genes, at the RNA level by in situ hybridization. However, de novo expression of HSP70 could be observed during fever (usually above 39°C) in only about half of the cases. No simple threshold model for inducibility of HSP70 in vivo could be applied. The HSP70-1 gene was induced more easily than HSP70-2 . Thus, heat-inducible HSP70 genes, including HLA-linked HSP70 genes, become expressed in human lymphocytes during fever, but not regularly.     

早期谷氨酰胺强化肠外营养对危重病患者体内热休克蛋白70的影响

目的探讨早期给予谷氨酰胺(Gln)强化肠外营养对危重病患者脏器功能的影响及其与预后的关系。方法选择本院急诊及神经外科重症加强治疗病房(NICU)收治的44例危重病患者,按随机原则分为常规治疗组和Gln治疗组,每组22例。两组患者均行肠内、外营养,同时Gln治疗组静脉注射Gln 0.4g·kg~(-1)·d~(-1),连用7d;观察两组患者治疗前后体内热休克蛋白70(HSP70)、Gln含量,机械通气时间、入住ICU时间及肝、肾功能不全的发生率。结果常规治疗组和Gln治疗组治疗前Gln、HSP70水平差异无显著性(P均>0.05);常规治疗组治疗后Gln、HSP70水平较治疗前稍有增加,但差异无显著性;而Gln治疗组治疗后Gln、HSP70水平均较治疗前显著升高,差异有显著性(P均<0.01),且两组治疗后Gln、HSP70水平比较差异均有显著性(P均<0.01)。Gln治疗组体内Gln浓度和HSP70含量的变化存在显著正相关(r=0.650 5,P=0.001)。两组机械通气时间和肝功能不全的发生率差异有显著性(P均<0.05),入住ICU的时间和肾功能不全的发生率则差异无显著性(P均>0.05)。结论给危重病患者早期肠外补充Gln能有效改善患者的预后,降低脏器功能不全的发生率,其机制可能与提高患者体内HSP70水平有关。     

热休克蛋白70与严重多发伤后早期继发性肝脏损害

目的 :探讨热休克蛋白 70 (HSP70 )在严重多发伤后肝脏继发性损害中的作用。方法 :成年雄性 Wistar大鼠 ,采用严重胸部撞击伤伴单侧股骨骨折多发伤模型。动态观察致伤后 2 4 h及用糖皮质激素受体阻断剂米非司酮后大鼠肝组织 HSP70、肝功能指标及致伤 2 4 h死亡率等变化。HSP70表达测定采用免疫印迹法 ,并进行计算机图像分析。结果 :伤后 HSP70在肝组织中表达伤后迅速增加 ,8h达到峰值 ,2 4 h仍维持在较高水平 ;但血清丙氨酸转氨酶 (AL T)、总胆红素 (TB)及白蛋白的改变无显著差异。使用糖皮质激素受体阻断剂米非司酮能使肝组织 HSP70表达明显增多 ;血清 AL T及 TB在伤后早期即有明显升高 (P均 <0 .0 1) ,白蛋白明显下降(P<0 .0 1) ;伤后 2 4 h动物死亡率明显增加 (P<0 .0 1)。结论 :HSP70可能参与了肝组织细胞抗损伤机制的启动 ,但 HSP70过高表达则可能对肝脏造成损害。 HSP70可作为创伤后肝损伤与抗损伤机制的一个重要指标。     

顺铂内耳中毒豚鼠耳蜗热休克蛋白70表达的研究

目的观察顺铂内耳中毒豚鼠耳蜗热休克蛋白70(HSP70)的表达特点。方法制备顺铂内耳中毒听力损伤模型,分别于第3天,第6天处死豚鼠,取耳蜗做石蜡包埋切片,进行HSP70免疫组化(SP法)染色,检测耳蜗HSP70的表达和分布特点。图像分析采用计算机图像分析系统。结果对照组中Corti器、血管纹、螺旋缘、螺旋神经节呈弱阳性。Ⅱ组中Corti器、血管纹、螺旋缘、螺旋神经节的HSP70表达增强。Ⅲ组中Corti器、血管纹、螺旋缘、螺旋神经节呈强阳性。Ⅲ组平均灰度值较对照组有显著性差异(P<0.01)。结论顺铂能够诱导HSP70在豚鼠耳蜗中的表达,使HSP70在豚鼠耳蜗中的表达增强。     

戊四氮致癫痫大鼠脑组织形态结构及热休克蛋白70表达的初步研究

目的：探讨癫痫的不同发作级别所造成的神经元损伤及热休克蛋白70（HSP70）的表达情况及其意义。方法：采用戊四氮致癫痫模型,随机分为生理盐水对照组和模型组,模型组按癫痫发作级别又分为Ⅰ、Ⅲ、Ⅴ级组。采用常规病理学染色观察各组海马区神经元的受损情况;免疫组织化学方法观察不同级别癫痫发作海马内HSP70表达的变化。结果：Ⅲ、Ⅴ级组与对照组和Ⅰ级组相比,海马结构内存在受损异常神经元,Ⅴ级组更明显（P〈0．05）。同时Ⅲ级组、Ⅴ级组海马各区的HSP70阳性细胞数亦逐渐增多（P〈0．05）。结论：戊四氮致癫痫模型中,随着发作级别的增加,海马结构内神经元损伤逐渐加重,HSP70的表达亦逐渐增加,提示海马结构HSP70的表达与癫痫发作严重程度有关。     

腺病毒介导的热休克蛋白70在神经元和胶质细胞中的抗缺氧研究

目的 探讨重组腺病毒介导的热休克蛋白70(HSP70)表达对神经元和胶质细胞缺氧/再复氧损伤的保护作用.方法 制备携带全长HSP70基因的重组腺病毒vAd-HSP70.感染体外培养的神经元和胶质细胞,检测靶细胞中外源性HSP70的表达.感染vAd-HSP70 24、48、72 h组和感染vAd-GFP对照组的细胞经缺氧/再复氧处理后,分别测定细胞活性、细胞培养上清液中乳酸脱氢酶(LDH)活性及线粒体和胞质内细胞色素C含量.结果 感染vAd-HSP70的神经细胞可检测到外源性HSP70基因表达.经缺氧/再复氧处理,感染vAd-HSP70组细胞活性较感染vAd-GFP对照组明显增强(P均＜0.05);感染vAd-HSP70 24、48、72 h组细胞培养上清液中LDH活性[(1 480±121)、(1 023士106)、(1 132±197)U/L]均明显低于感染vAd-GFP对照组[(1 976±190)U/L],线粒体中细胞色素C含量(0.986±0.012、1.028±0.007、1.014±0.008)均明显高于感染vAd-GFP对照组(0.970±0.003),而胞质中的细胞色素C含量(0.987±0.008、0.960±0.005、0.964±0.003)则低于感染vAd-GFP对照组(1.011±0.005,P＜0.05或P＜0.01),其中以感染48 h最为理想(P均＜0.01).结论 腺病毒介导的外源性HSP70表达可保护神经元和胶质细胞抵抗缺氧/再复氧损伤,具有明确的细胞保护作用.     

Constitutively expressed small heat shock protein LsHsp21.5 not only enhances heat tolerance but also helps to maintain reproduction in female Laodelphax striatellus

Coping with stressful conditions and maintaining reproduction are two key biological processes that affect insect population dynamics. Small heat shock proteins (sHSPs) are involved in the stress response and the development of insects. The sHsp gene Laodelphax striatellus (Hemiptera: Delphacidae) sHsp 21.5 (LsHsp21.5) showed constitutive, stage- and organ-specific expression in L. striatellus, a pest that damages cultivated rice in east Asia. The expression of LsHsp21.5 was highest in the ovary, with 43.60, 12.99 and 1.45 time higher expression here than in the head, gut and female fat bodies, respectively. The expression of this gene was weakly affected by heat or cold shock. The gene provided in vitro protection against heat damage to malate dehydrogenase and in vivo protection against heat stress in Escherichia coli (Enterobacteriales: Enterobacteriaceae) BL21(DE3) and L. striatellus. Moreover, L. striatellus reproduction decreased by 1.85 times when the expression of LsHsp21.5 was inhibited by RNA interference. The expression of some genes related to reproduction, such as the homologous gene of chorion protein, also declined. These results suggest that LsHsp21.5 expression not only protects other proteins against stress but also helps maintain the stable expression of some reproduction-related genes under non-stressful conditions, with impacts on L. striatellus fecundity.     

脑脊液热休克蛋白70水平对小儿中枢神经系统感染的鉴别诊断价值

目的 探讨脑脊液(CSF)中热休克蛋白70(HSP70)的变化及其在小儿中枢神经系统感染中的诊断价值.方法 采用蛋白质免疫印迹技术检测13例化脓性脑膜炎(化脑组)、38例病毒性脑膜炎(病脑组)、7例结核性脑膜炎(结脑组)及46例非中枢神经系统感染患儿(对照组)CSF中HSP70水平.常规生化检测CSF的细胞总数、白细胞数、乳酸脱氢酶(LDH)、蛋白定量、腺苷脱氨酶、葡萄糖、压力及氯(Cl-) 水平.结果 化脑组(76.61±27.69)、病脑组(33.65±16.93)及结脑组(65.85±33.16)的HSP70水平均高于对照组(23.28±19.77),差异有显著性(P＜0.05或P＜0.01);化脑组及结脑组HSP70水平均高于病脑组(P均＜0.01);化脑组与结脑组之间HSP70水平差异无显著性(P＞0.05).相关性分析显示:HSP70水平增高程度与CSF的细胞总数(r=0.298,P=0.002)、白细胞数(r=0.274,P=0.005)、LDH(r=0.322,P=0.001)、蛋白定量(r=0.629,P＜0.001)、腺苷脱氨酶水平(r=0.363,P=0.001)均呈显著正相关,与CSF中葡萄糖水平呈显著负相关(r=-0.443,P＜0.001),与CSF压力(r=0.001,P=0.993)及Cl-水平(r=0.148,P=0.133)无相关性.结论 小儿中枢神经系统感染时,CSF中HSP70增高;检测CSF中HSP70水平有助于化脑、结脑与病脑的鉴别诊断.     

热休克蛋白70与激素受体在男性乳腺发育和男性乳癌中的表达及其意义

目的 探讨热休克蛋白７０（ＨＳＰ７０）与激素受体（ＥＲ、ＰＲ）在男性乳腺发育和男性乳癌右的表达情况及分布状态。方法 应用免疫组织化学Ｓ－Ｐ法对２５例男性乳腺发育和１例男性乳癌进行检测。结果 在２５例男性乳腺发育病变中,ＨＳＰ７０阳性率为７２．０％（１８例）;ＥＲ、ＰＲ阳性率分别为６８．０％和６０．０％（１７例和１５例）;１例男性乳癌三种染色均阳性,ＨＳＰ７０阳性细胞在男性乳腺发育和男性乳癌组织中,细胞核、细胞浆均有表达,ＥＲ、ＰＲ阳性细胞主要位于细胞核、尤其以男性乳癌细胞表达更显著,结论 ＨＳＰ７０可作为男性乳腺发育及乳癌细胞增殖的指标之一。雌、孕激素受体推移与男性乳腺发育及男性乳癌的发生可能有关,尚需进一步研究。