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Characterization of T-complex polypeptide 1 (TCP-1) from the Chilo suppressalis HSP60 family and its expression in response to temperature stress

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Journal of Integrative Agriculture 2018年第5期17卷 1032-1039页

作者： YU Tong-ying lu ming-xing CUI Ya-dong Biology and Food Engineering School Fuyang Normal University School of Horticulture and Plant Protection & Institute of Applied Entomology Yangzhou University

Many proteins require assistance from molecular chaperones at various stages to attain correctly folded states and functional conformations during protein synthesis. In this study, the gene encoding T-complex polypeptide 1（TCP-1）, which belongs to the heat shock protein 60（HSP60） family, was isolated and characterized from the rice stem borer, Chilo suppressalis, by RACE and q PCR, respectively. The full-length c DNA of Tcp-1 was 2 144 bp and encoded a 1 635-bp ORF; the deduced translational product contained 545 amino acids with 5′-and 3′-UTRs and an isoelectric point of 5.29. Cluster analysis confirmed that the deduced amino acid sequence shared high identity（60–99%） with TCP-1 from other insects. To investigate Tcp-1 expression in response to abiotic stress, q PCR was used to analyze expression levels of Tcp-1 m RNA in C. suppressalis larvae exposed to temperatures ranging from –11 to 43°C. With respect to heat shock, Tcp-1 expression was higher than the control after a 2-h exposure to 30 and 36°C and declined at 39 and 43°C. Difference in Tcp-1 expression was observed at temperatures ranging from –11 to 27°C. q PCR analyses revealed that Tcp-1 expression was the highest in hindgut tissue as compared to heads, epidermis, fat body, foregut, midgut, and malpighian tubules. Our results indicated that Tcp-1 expression was differentially expressed in C. suppressalis tissues, and was impacted by temperature stress.

关键词： TCP-1 Chilo suppressalis expression analysis temperature stress insect tissues

No evidence for an effect of Wolbachia on mtDNA variation and evolution in natural populations of Sesamia inferens(Lepidoptera: Noctuidae)

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Journal of Integrative Agriculture 2019年第5期18卷 1050-1063页

作者： TANG Xiao-tian XU Jing lu ming-xing DU Yu-zhou School of Horticulture and Plant Protection & Institute of Applied Entomology Yangzhou University Department of Entomology Texas A&M University Joint International Research Laboratory of Agriculture and Agri-Product Safety Ministry of Education/Yangzhou University

Wolbachia are widespread maternally-inherited endosymbiotic bacteria that infect numerous *** study represents a thorough survey of the Wolbachia infection patterns in the pink stem borer,Sesamia inferens(Walker),an important rice pest in China,based on nucleotide comparisons for the surface protein(wsp)and cell division protein(ftsZ)*** effects of Wolbachia on mtDNA variation and evolution of *** were also *** we identified six genetically diverse strains,we found infections to be infrequent,with only 7.8%of hosts infected,and identified geographical differences in infection rates between southern and northern *** indexes(haplotype diversity(Hd),nucleotide diversity(π)and number of variable sites(S)of mtDNA in infected populations were not significantly lower or higher than that in the uninfected ***,there was no association between Wolbachia infection status and phylogeny of mtDNA *** of molecular variance(AMOVA)showed that significant differentiation mainly existed within groups rather than among the ***,using Tajima’s D and Fu’s F values,the mtDNA genes did not deviate significantly from neutral *** together these results indicate that currently there were no effects of Wolbachia infection on host mtDNA variation and evolution in ***.

关键词： Wolbachia Sesamia inferens effect mtDNA prevalence

Characterization of two novel heat shock protein 70s and their transcriptional expression patterns in response to thermal stress in adult of Frankliniella occidentalis (Thysanoptera:Thripidae)

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Journal of Integrative Agriculture 2018年第5期17卷 1023-1031页

作者： QIN Jing GAO Peng ZHANG Xiao-xiang lu ming-xing DU Yu-zhou School of Horticulture and Plant Protection & Institute of Applied Entomology Yangzhou University

Heat shock protein 70（HSP70） is one of the most important members in the heat shock protein family, and plays important roles in the thermotolerance of insect. To explore the molecular mechanism of thermotolerance of Frankliniella occidentalis adults, the difference in the expression of HSP70s in F. occidentalis male or female adults under the thermal stress was studied under the laboratory conditions. Two full length c DNAs of HSP70s gene（Fohsc704 and Fohsc705） were cloned from F. occidentalis by using RT-PCR and RACE. The genomic sequence was demonstrated by genomic validation, and the position and size of the intron were analyzed by sequence analysis of c DNA. Real-time PCR was used to analyze the HSP70 expression patterns. The c DNA of Fohsc704 and Fohsc705 possessed 2 073 and 1 476 bp which encoded 690 and 491 amino acids（aa） with a calculated molecular weight of 75 and 54 k Da, respectively. Four introns in Fohsc704 and six introns in Fohsc705 protein were found. However, the HSP70 protein sequences in our study were ended with EKKN and GIFL, which were different from the reported Fo HSP70s. Various expression patterns of Fohsc704 and Fohsc705 were found in both genders of F. occidentalis under thermal stress. The expression of Fohsc704 and Fohsc705 reached to the highest level at –12 and –8°C in male adults, respectively, and Fohsc705 expressed the highest level at 33°C in female adults. In conclusion, HSP70s of F. occidentalis in our study are novel heat shock proteins. There were difference in expression patterns of the two hsc70s in genders of F. occidentalis, and the two HSP70s play important roles in the thermotolerance of F. occidentalis.

关键词： Frankliniella occidentalis HSP70 temperature gene cloning expression

Genes encoding heat shock proteins in the endoparasitoid wasp,Cotesia chilonis,and their expression in response to temperatures

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Journal of Integrative Agriculture 2018年第5期17卷 1012-1022页

作者： PAN Dan-dan CAO Shuang-shuang lu ming-xing HANG San-bao DU Yu-zhou School of Horticulture and Plant Protection & Institute of Applied Entomology Yangzhou University Joint International Research Laboratory of Agriculture and Agri-Product Safety Ministry of Education/Yangzhou University

Five genes encoding heat shock proteins（HSPs）, Cchsp40, Cchsp60, Cchsp70, Cchsc70 and Cchsp90, were cloned and sequenced from Cotesia chilonis using RT-PCR and RACE. The c DNA sequences of Cchsp40, Cchsp60, Cchsp70, Cchsc70 and Cchsp90 were 1 265, 2 551, 2 094, 2 297 and 2 635 bp in length, respectively, with a molecular weight（MW） of 39.1, 60.6, 71.45, 70.19 and 82.92 k Da, respectively. The predicted amino acid sequences of these proteins showed high similarities with published HSPs of other insects in Hymenoptera. Analysis of genomic DNAs indicated that Cchsp40, Cchsp60, Cchsp70 and Cchsp90 lacked introns, but Cchsc70 contained an intron. The results also suggested that CcH SP40 in C. chilonis was the Type II HSP40, Cc HSP60 was a member of the mitochondrial HSP60 family, and Cc HSP90 was a part of cytoplasmic HSP90 A family. Expression patterns varied in the five Cchsps in response to temperature. Expression of Cchsp40 and Cchsp60 was induced significantly by cold but not heat stress. Cchsp70 and Cchsc70 showed similar response to the thermal stress and could be induced by both cold and heat, but their expression levels were consistently lower than that of Cchsp40 and Cchsp60. Cchsp90 could be induced by heat stress and mild cold, but not cold stress. In addition, the results demonstrated Cchsc70 might be constitutive and inducible protein that was expressed during normal cell functioning and also up-regulated in response to stressful stimuli while Cchsp70 was solely inducible protein induced by temperature changes. Overall, results generated from this study could significantly advance the understanding of Cchsps in response to temperature and provide important biological information for C. chilonis insects that reared under different temperatures.

关键词： Cotesia chilonis HSPs genomic structure temperature expression

Effect of Rapid Cold Hardening on the Cold Tolerance of the Larvae of the Rice Stem Borer,Chilo suppressalis(Walker)

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Agricultural Sciences in China 2008年第3期7卷 321-328页

作者： QIANG Cheng-kui DU Yu-zhou YU Ling-ya CUI Ya-dong ZHENG Fu-shan lu ming-xing Institute of Applied Entomology Yangzhou University Yangzhou 225009 P.R.China College of Life Sciences Fuyang Teachers College Fuyang 236032 P.R.China College of Resources and Environment South China Agricultural University Guangzhou 510642 P.R.China 不详

The effects of rapid cold hardening （RCH） on the cold tolerance of the last instar larvae of Chilo suppressalis （Walker） were evaluated for the first time. The discriminating temperature, induction, detection, duration and extent of RCH of the larvae in the laboratory were tested, and the supercooling points （SCPs） and the contents of water and lipid of the larvae after RCH treatment were determined, respectively. The results showed that the discriminating temperature of the larvae was about -21℃. Mean survival rates of the larvae which exposed to either 0 or 5℃ for 2 and 4 h before exposure to the discriminating temperature for 2 h were significantly higher than those of the control groups （P 〈 0.05）. Moreover, the highest survival rate appeared in the larvae after 0℃ for 4 h treatment. The protection against low temperature gained by RCH at 0℃ for 4 h was rapidly lost on return to 28℃. Mean survival rates of RCH larvae were significantly higher than those of non-acclimated （NACC） larvae and acclimation （ACC） larvae when they were exposed to the discriminating temperature for 2 or 4 h （P〈 0.05）. Moreover, the rates of NACC, ACC, RCH and ACC ＋ RCH larvae from 2 to 6 h to the discriminating temperature resulted in a significant decline. The values of SCPs and the contents of lipid of the larvae which exposed to either 0 or 5℃ for 2 and 4 h showed no significant difference at 0.05 level compared to those of the control groups. But the contents of water in the larvae were obviously decreased. Therefore, it could be concluded that RCH could enhance cold tolerance and affect partly physiological and biochemical components of the larvae of C. suppressalis, but the underlying mechanisms needs to be further explored.

关键词： Chilo suppressalis cold tolerance rapid cold hardening supercooling point survival rate

Absorption characteristics of the total alkaloids from Mahonia bealei in an in situ single-pass intestinal perfusion assay

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Chinese Journal of Natural Medicines 2014年第7期12卷 554-560页

作者： SUN Yu-He HE Xin YANG Xiao-Lin DONG Cui-Lan ZHANG Chun-Feng SONG Zi-Jing lu ming-xing YANG Zhong-Lin LI Ping State Key Laboratory of Natural Medicines China Pharmaceutical University School of pharmacy Nanjing University of Chinese Medicine Guangdong Provincial Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research Jinan University The People's Hospital of Zhangqiu City

AIM: To investigate the absorption characteristics of the total alkaloids from Mahoniae Caulis(TAMC) through the administration of monterpene absorption enhancers or protein inhibitors. METHOD: The absorption behavior was investigated in an in situ single-pass intestinal perfusion(SPIP) assay in rats. RESULTS: The intestinal absorption of TAMC was much more than that of a single compound or a mixture of compounds(jatrorrhizine, palmatine, and berberine). Promotion of absorption by the bicyclic monoterpenoids(borneol or camphor) was higher than by the monocyclic monoterpenes(menthol or menthone), and promotion by compounds with a hydroxyl group(borneol or menthol) was higher than those with a carbonyl group(camphor or menthone). The apparent permeability coefficient(Papp) of TAMC was increased to 1.8-fold by verapamil, while it was reduced to one half by thiamine. The absorption rate constant(Ka) and Papp of TAMC were unchanged by probenecid and pantoprazole. CONCluSION: The intestinal absorption characteristics of TAMC might be passive transport, and the intestinum tenue was the best absorptive site. In addition, TAMC might be likely a substrate of P-glycoprotein(P-gp) and organic cation transporters(OCT), rather than multidrug resistance protein(MRP) and breast cancer resistance protein(BCRP). Compared with a single compound and a mixture of compounds, TAMC was able to be absorbed in the blood circulation effectively.

关键词： Mahoniae Caulis Total alkaloids In situ single-pass intestinal perfusion Absorption enhancers Protein inhibitors

下伏空洞地基极限承载力与破坏模式上限有限元法

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岩土力学 2017年第1期38卷 229-236,246页

作者：赵明华卢晓明张锐湖南大学岩土工程研究所湖南长沙410082

地基极限承载力的确定对于下伏空洞地区基础设计和施工具有重要意义。采用上限有限元法对地基破坏时的极限状态进行数值模拟。首先,简要介绍上限有限元法的基本原理与计算过程;进而提出合理的计算假定,建立可考虑多种影响因素的计算模型... 详细信息

地基极限承载力的确定对于下伏空洞地区基础设计和施工具有重要意义。采用上限有限元法对地基破坏时的极限状态进行数值模拟。首先,简要介绍上限有限元法的基本原理与计算过程;进而提出合理的计算假定,建立可考虑多种影响因素的计算模型;其次,利用上限有限元法计算出多种工况下的地基极限承载力上限解,按Tergaghi建议的地基承载力公式,得到承载力系数N_c、N_q、N_g,并对其影响因素进行分析;最后,根据大量的能量耗散图及速度场图提出3类典型的破坏模式,并对其影响因素进行分析。研究表明:随着内摩擦角j的增大,N_c和N_q均增大,而当j较大时,N_g才出现正数,且随着j增大而增大;随着空洞顶板厚度与基础宽度之比H/B的增大,N_c和N_q均增大。而当j较小时,N_g随着H/B的增大而减少;当j较大时,N_g随着H/B的增加而增加。N_c、N_q和N_g均随着D/B的增大而减少。下伏空洞地基存在3种典型的破坏模式:冲切破坏模式、冲切剪压破坏模式和Prandtl破坏模式。

关键词：空洞上限法有限元承载力系数破坏模式

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富水泥化软岩增稠试验研究

硅酸盐通报 2017年第8期36卷 2614-2618,2624页

作者：张彬鲁明星辽宁工程技术大学土木与交通学院阜新123000

富水软岩井筒掘进过程中,弱胶结软岩遇水泥化将形成具有流动特性的泥浆,导致工作面掘进和排矸困难,恶化工作面环境,增加工人劳动强度,严重影响施工速度。为有效解决此问题,提出在工作面泥浆中参入适量的高分子吸水性树脂、纤维素和多种... 详细信息

富水软岩井筒掘进过程中,弱胶结软岩遇水泥化将形成具有流动特性的泥浆,导致工作面掘进和排矸困难,恶化工作面环境,增加工人劳动强度,严重影响施工速度。为有效解决此问题,提出在工作面泥浆中参入适量的高分子吸水性树脂、纤维素和多种矿物质原料以增强泥浆的稠度,便于排矸。首先通过吸水率试验,研究高分子吸水性树脂、纤维素和多种矿物质原料等材料与水有关的性质;其次利用水泥跳桌流动度仪对掺入上述材料的流砂泥浆的稠度进行检测,研究高分子吸水性树脂、纤维素和多种矿物质原料对流砂泥浆稠度的影响规律,确定合适的吸水增稠材料以及配比。研究结果表明掺入高分子吸水树脂对增加流砂泥浆的稠度效果最好,纤维素次之,矿物质材料对流砂稠度的影响,远不如高分子吸水树脂和纤维素。

关键词：软岩矿井建设聚丙烯酸盐纤维素矿物质原料

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针刺耐受中枢机制的研究进展

针刺研究 2016年第6期41卷 550-555页

作者：崔璐莹丁明星华中农业大学动物医学院武汉430070

电针具有良好的镇痛效应。然而,反复多次电针会引起电针镇痛效应下降,导致电针耐受,严重影响电针的临床疗效。近年来关于电针耐受机制的研究取得了一定进展,主要包括以下方面:(1)电针耐受实验动物模型的建立,包括急性与慢性耐受;(2)电... 详细信息

电针具有良好的镇痛效应。然而,反复多次电针会引起电针镇痛效应下降,导致电针耐受,严重影响电针的临床疗效。近年来关于电针耐受机制的研究取得了一定进展,主要包括以下方面:(1)电针耐受实验动物模型的建立,包括急性与慢性耐受;(2)电针与吗啡的交叉耐受;(3)抗阿片类物质,如胆囊收缩素、孤啡肽和血管紧张素Ⅱ等,参与电针耐受;(4)其他参与电针耐受的物质,包括谷氨酸受体及转运体、5-羟色胺、去甲肾上腺素等;(5)参与电针耐受的细胞内第二信使,包括环磷酸腺苷、环磷酸鸟苷和Ca2+。目前,电针耐受的研究仍缺乏深入性,对其分子机制和信号通路的研究十分必要。

关键词：电针镇痛电针耐受中枢机制神经递质神经调质