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Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

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Journal of Oceanology and Limnology 2020年第2期38卷 480-489页

作者： CHEN Xiaojuan ZHANG Xiaoqi SUN Yun TU Zhigang CAO Zhenjie WANG Shifeng zhou yongcan State Key Laboratory of Marine Resource Utilization in South China Sea Hainan UniversityHaikou 570228China Hainan Academy of Ocean and Fisheries Sciences Haikou 570228China

Trachinotus blochii is one of the important commercial fish *** this study,we aim to confirm the reliability reference genes in *** during different bacterial challenge through quantitative real-time PCR(qRT-PCR).The expression of the seven selected genes in four immune organs(i.e.,spleen,kidney,intestine,and gill)stimulated with Vibrio harveyi,Edwardsiella tarda,and Streptococcus agalactiae were determined by *** PCR data was analyzed using the geNorm and NormFinder *** results showed the selection of the internal controls should be tissue specific when studying gene expression in response to bacterial *** 48 h of stimulation with ***,geNorm ranked EF1 A/Actin,18 S rRNA/B2M,UBCE/B2M,and 18 S rRNA/B2M,as the most stably expressed genes in spleen,kidney,intestine,and gill,*** 48 h of stimulation with ***,geNorm ranked 18 S rRNA/EF1 A,18 S rRNA/B2M,B2M/RPL13,and 18 S rRNA/EF1 A,as the most stably expressed genes in spleen,kidney,intestine,and gill,*** 48 h of stimulation with ***,18 S rRNA/EF1 A,18 S rRNA/B2 M,B2 M/Actin,and 18 S rRNA/B2M were ranked as the most stably expressed genes in spleen,kidney,intestine,and gill,*** to the results analyzed by geNorm,reference genes received similar rankings when using NormFinder *** results showed that the reference genes appeared to be not only tissue specific,but also specific to the infecting species of *** one gene is preferred when *** were infected by bacteria,18 S rRNA,B2M,B2M,18 S rRNA may be used in spleen,kidney,intestine,and gill,respectively.

关键词： Trachinotus blochii housekeeping gene expression stability reference gene

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Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis

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Journal of Oceanology and Limnology 2021年第5期39卷 1985-1999页

作者： Xiaojuan CHEN Yun SUN Panpan ZHANG Jianlong LI Haiping LI Caoying WEI Zhenjie CAO yongcan zhou State Key Laboratory of Marine Resource Utilization in South China Sea Hainan UniversityHaikou 570228China Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province Haikou 570228China Department of Aquaculture College of Marine SciencesHainan UniversityHaikou 570228China

Humpback grouper Cromileptes altivelis is one commercial fish with considerable economic *** determine the expression stabilities of six commonly used internal reference genes in *** challenged by Vibrio harveyi and viral nervous necrosis virus(VNNV)through quantitative real-time PCR(qRT-PCR),the expression levels of selected genes in five immune organs stimulated with pathogenic infection were carefully evaluated using algorithms of geNorm,NormFinder,and *** results show that the expre ssion stabilities of the six candidate inte rnal reference genes were diffe re *** no rmal physiological conditions,RPL13 were identified as the most stably expressed genes among five different immune organs(liver,spleen,kidney,intestine,and gill).After *** stimulation,RPL13,RPL13,EF1 A,RPL13,and EF1 A were identified by geNorm,NormFinder,and BestKeeper as the most stable genes in liver,spleen,kidney,intestine,and gill,*** these three algorithms suggested that under stimulation of VNNV,RPL13,EF1 A,Actin,RPL13,and Actin were as the most stable genes in liver,spleen,kidney,intestine,and gill,*** results suggest that specific experiment conditions and tissue types shall be considered when selecting the reference genes in qRT-PCR *** study provided a solid foundation for future studies on gene expression of *** under different conditions.

关键词： Cromileptes altivelis reference gene expression stability pathogenic infection

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Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers Plectropomus leopardus

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Journal of Oceanology and Limnology 2023年第3期41卷 1084-1099页

作者： Han ZHANG Haoran YANG Pengsuo LI Na ZHAO Xiaojuan CHEN yongcan zhou Zhenjie CAO Yun SUN State Key Laboratory of Marine Resource Utilization in South China Sea Hainan UniversityHaikou 570228China Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province Haikou 570228China Department of Aquaculture College of Marine SciencesHainan UniversityHaikou 570228China

Leopard coral groupers(Plectropomus leopardus),commercially bred in South China,are a significant economical fish *** this study,by means of quantitative real-time PCR(qRT-PCR)technology,we assessed the stability of six common reference genes expression and selected the appropriate reference genes in leopard coral groupers with or without Vibrio harveyi stimulation at different time *** data produced by qRT-PCR was handled via the geNorm,NormFinder,and BestKeeper *** results revealed all the examined reference genes had manifest tissue-specific expression in different *** to *** stimulation,RPL13 gene was the appropriate reference gene among eleven tissue types(blood,spleen,hepatopancreas,kidney,stomach,gill,heart,skin,muscle,intestine,brain)in leopard coral *** *** stimulation,the most reliable reference genes varied from tissue to tissue and were closely hinged upon different time *** 6-h post-bacterial injection,the appropriate reference genes in hepatopancreas,spleen,kidney,and gill were Actin,B2M,UBCE,and Actin,*** 9-and 12-h post-bacterial injection,the appropriate reference genes in hepatopancreas,spleen,kidney,and gill were RPL13,Actin,Actin,and Actin,*** one reference gene was preferable,RPL13,Actin,Actin,and Actin could be selected as the reference gene in hepatopancreas,spleen,kidney,and gill of leopard coral groupers after *** infection,*** profiles of two target genes(IL-6 and NK-lysin)were used to further validate reliability of these selected appropriate *** study will lay a solid foundation for the future research on qRTPCR analysis of gene expression in leopard coral groupers.

关键词： Plectropomus leopardus reference genes Vibrio harveyi stimulation

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Studies on structure and function of digestive system of Cymatium pileare

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Acta Oceanologica Sinica 2001年第2期20卷 261-270页

作者： zhou yongcan, Zeng Shuixiang, Su Yongquan (1. Fishery Department, Hainan University, Haikou 570228, China 2. Oceanology Department, Xiamen University, Xiamen 361005, China) Fishery Department Hainan University Oceanology Department Haikou 570228 China

The digestive system of Cymatium pileare consists of buccal cavity, oesophagus, stom- ach, intestine, anus, salivary glands and liver. There is a contractible 'Argobuccinum' form proboscis in the anterior extremity of its body, and the buccal cavity lies in the anterior part of the proboscis, with a pair of jaws and a piece of radular ribbon. The jaws have many neatly arraned sickle- shaped sharpen denticles. Its dentition formula of radula is 2- (7) 1' (5-6) I (5-6) .1 (7) .2. In the middle of oesophagus, there is an expanded crop, which has the function of temporary food stor- age and preliminarily digesting food. The stomach is U-shaped and there are 2 entrances for liver ducts in its ventral side. The intestine is short and can be divided into 2 parts based on the structure of epithelia of their inner walls. The inner wall of foreintestine is stratified columnar epithelium, but the inner wall of hindintestine is pesudostratified columnar epithelium. The salivary gland of C. pileare is toxic gland and the toxin extracted from it plays a great role on its predation. The liver of C. pileare is compound tubuloacinar gland with mitochondrion, rough endoplasmic reticulum, Golgi comples and lysosome in its glandular cells.

关键词： Cymatium pileare, digestive system, structure function

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Dietary administration of Bacillus subtilis HAINUP40 enhances growth,digestive enzyme activities,innate immune responses and disease resistance of tilapia,Oreochromis niloticus

Dietary administration of Bacillus subtilis HAINUP40 enhance...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Shifeng Wang Haitian Liu Yan Cai Wei Yuan yongcan zhou Key Laboratory of Tropical Biological Resources of Ministry of Education College of Marine SciencesHainan University

The probiotic properties of Bacillus subtilis HAINUP40 isolated from the aquatic environment,and the effects of dietary administration of *** HAINUP40 on the growth performance,intestinal probiotic recovery,digestive enzyme activities,innate immunity and disease resistance of tilapia(Oreochromis niloticus) were *** probiotic properties investigated include tolerance to simulated gastrointestinal stress,auto-aggregation,cell surface hydrophobicity and extracellular enzyme *** cell number of *** changed little after 4 h in simulated gastric fluid at pH = 2.0,3.0,4.0 and small intestinal fluid at pH = *** HAINUP40 revealed a strong auto-aggregation property(34.6-87.0%) after a 24 h incubation *** exhibited significant cell surface hydrophobicity in xylene(28.8%) and chloroform(41.3%) and produced extracellular proteases and *** tilapia(mean weight = 95.02 ± 7.65 g) were fed with a diet containing 10cfu/g *** HAINUP40,their final body weight,percent weight gain(PWG),specific growth rate(SGR),total antioxidant capacity(T-AOC) and serum superoxide dismutase(SOD) increased significantly(p<0.05) after 8weeks;feed conversion rate(FCR) is significantly lower(p < 0.05) after 8 weeks;the protease and amylase activity in the digestive tract increased significantly(p<0.05) after 4 and 8 weeks;and respiratory bursts and serum lysozyme activity increased significantly(p< 0.05) after 2 ***,being challenged with pathogenic Streptococcus agalactiae for 2 weeks,the relative percent survival(RPS%) is 52.94%.The results of this study strongly suggest that dietary supplement of *** HAINUP40 can effectively enhances the growth performance,immune response,and disease resistance of Nile tilapia.

关键词： Bacillus subtilis HAINUP40 Tilapia(Oreochromis niloticus) Growth Digestive enzyme activity Innate immunity Protection

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Molecular cloning and characterization of two types of IκBα homolog in orange-spotted grouper, Epinephelus coioides

Molecular cloning and characterization of two types of IκB...

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中国水产学会鱼病专业委员会2013年学术研讨会

作者： Ren Gao Youhua Huang Liya Guan Shina Wei Xiaohong Huang yongcan zhou Qiwei Qin State Key Laboratory Breeding Base for Sustainable Exploitation of Tropical Biotic Resources College of Marine Science Hainan University Key Laboratory of Tropical Marine Bio-resources and Ecology South China Sea Institute of OceanologyChinese Academy of Sciences

IκBa is a member of the primary regulators of NF-κB,which can inhibit NF-κB activityby blocking the NF-κB in an inactive state in the *** this study,two types of IκBαhomolog,EcIκBα1and EcIκBα2,were cloned and characterized from orange-spotted grouper,Epinephelus *** complete length of EcIκBα1cDNA is 1629 bp and encodes a putativeprotein of 318 amino acids sharing 85%identity to IκBαof Takifugu rubripes,while the fulllength of EcIκBα2cDNA is 1441 bp,which encodes a protein of 308 residues with 89%identity toIκBαof Siniperca *** with the total 4092 bp genomic DNA of EcIκBα1,EcIκBα2is only 2419 bp *** EcIκBα1and EcIκBα2consist of 6 exons and 5 introns,which is similarto IκBαof Takifugu rubripes and Siniperca *** acid sequence analysis showed thatboth EcIκBα1and EcIκBα2protein contained a same conserved degradation motif DSGLDS intheir N-terminal region and a PEST sequence in the C-terminal region ***κBα1hadsix ankyrin repeats,while EcIκBα2had five ankyrin repeats and another PEST domain whichcontains a NES sequence in ***-PCR revealed that the transcripts of EcIκBα1andEcIκBα2were abundant in spleen but less in skin and ***-luciferase reporter assaysuggested that both EcIκBα1and EcIκBα2were able to inhibit EcC-Rel NF-κB and Ecp65 NF-κBtranscriptional activity in EAGS *** localization analysis showed that EcIκBα1wasdistributed predominantly in the cytoplasm,while EcIκBα2was distributed throughout both thenucleus and the *** expression level of EcIκBα1and EcIκBα2in cells stimulated bylipopolysaccharide(LPS),Vibrio alginolyticus and SGIV revealed that EcIκBα2might be moreeasily to be synthesized and degradated than EcIκBα1during pathogen *** findingssuggested that both EcIκBα1and EcIκBα2involved in grouper immune defense against invasion ofviral and bacterial pathogens by regulating the EcC-Rel NF-κB and Ecp65 NF-κB,and EcIκBα2might play a more significant role d

关键词： Epinephelus coioides EcIκBα 1 and EcIκBα 2 inhibitory activity fish immunity

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节能发电调度模式下的AGC机组调配策略研究

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浙江电力 2019年第5期38卷 103-108页

作者：周永灿吴英俊中国南方电网电力调度控制中心广州510623 河海大学能源与电气学院南京211100

AGC(自动发电控制)机组的调配是辅助服务中的一项重要内容。由于AGC服务与有功电力之间具有较强的耦合性,节能发电调度下如何协调AGC和有功电力是一个值得研究的问题。利用层次分析法对AGC机组的各方面要素进行量化,并通过算例与发电序... 详细信息

AGC(自动发电控制)机组的调配是辅助服务中的一项重要内容。由于AGC服务与有功电力之间具有较强的耦合性,节能发电调度下如何协调AGC和有功电力是一个值得研究的问题。利用层次分析法对AGC机组的各方面要素进行量化,并通过算例与发电序位表对所提两种方案进行对比分析,为节能发电调度模式下AGC机组的调配提供参考。

关键词：节能发电调度 AGC 层次分析法负荷备用

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海南岛海水珍珠贝养殖区嵌线螺的种类、分布及危害

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热带海洋 1999年第1期18卷 83-89页

作者：周永灿潘金培中国科学院南海海洋研究所广州510301

对海南岛３个海水珍珠贝养殖区养殖贝笼内的嵌线螺及其危害的研究表明，养殖区珍珠贝养殖笼内嵌线螺的发生具有明显的季节性和地域性：低温的１２月至翌年２月份没有发生，高温的６－７月份为发生高峰期；水温较低的北部水域出现少，水... 详细信息

对海南岛３个海水珍珠贝养殖区养殖贝笼内的嵌线螺及其危害的研究表明，养殖区珍珠贝养殖笼内嵌线螺的发生具有明显的季节性和地域性：低温的１２月至翌年２月份没有发生，高温的６－７月份为发生高峰期；水温较低的北部水域出现少，水温较高的南部水域出现多。在各实验区的养殖贝笼内全年共发现嵌线螺１０种，其中毛嵌线螺Ｃｙｍａｔｉｕｍｐｉｌｅａｒｅ和环沟嵌线螺Ｃ．ｃｉｎｇｕｌａｔｕｍ为优势种。嵌线螺的捕食是造成海南岛南部海水养殖珍珠贝死亡的主要因素。

关键词：嵌线螺珍珠贝危害海南岛种类珍珠贝

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精神科首诊早期肺癌脑转移1例

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临床心身疾病杂志 2021年第2期27卷 171-172页

作者：周湧灿浙江省立同德医院浙江杭州310012

脑转移肿瘤是成人中最为常见的颅内肿瘤,也是患者预后不良的重要原因,其中以肺癌发生脑转移最为多见。肺癌脑转移患者可因起病时肺部症状隐蔽且无特征性表现而被忽视,并因癌细胞转移颅内侵犯及压迫脑组织出现神经及精神症状等脑先行表... 详细信息

脑转移肿瘤是成人中最为常见的颅内肿瘤,也是患者预后不良的重要原因,其中以肺癌发生脑转移最为多见。肺癌脑转移患者可因起病时肺部症状隐蔽且无特征性表现而被忽视,并因癌细胞转移颅内侵犯及压迫脑组织出现神经及精神症状等脑先行表现而就诊。本文报道了以精神障碍症状为首发症状的早期肺癌脑转移患者1例,以提高对本病的重视。

关键词：肺癌脑转移早期诊断精神障碍

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毛嵌线螺的研究Ⅰ.繁殖及幼虫发生特征

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海洋学报 2000年第2期22卷 97-104页

作者：周永灿陈国华潘金培海南大学水产系海口570228 中国科学院南海海洋研究所广州510301

毛嵌线螺（Ｃｙｍａｔｉｕｍｐｉｌｅａｒｅ）的繁殖过程包括交配、产卵、孵育及幼虫从卵囊中游出等４个阶段．交配一般在产卵前１０～１５ｄ进行，产卵后母体并不离开卵群，而是持续静伏在卵群的开口上进行孵育，直至其中的幼虫离开... 详细信息

毛嵌线螺（Ｃｙｍａｔｉｕｍｐｉｌｅａｒｅ）的繁殖过程包括交配、产卵、孵育及幼虫从卵囊中游出等４个阶段．交配一般在产卵前１０～１５ｄ进行，产卵后母体并不离开卵群，而是持续静伏在卵群的开口上进行孵育，直至其中的幼虫离开卵囊．温度和盐度对毛嵌线螺受精卵的发育有重要影响，在２２～３０℃的适宜温度区域内，将受精卵孵育成能自由生活的晚期面盘幼虫需４３８日度左右的总积温．毛嵌线螺受精卵的发育与其他前鳃类腹足动物相似，包括卵裂、囊胚期、原肠期及面盘幼虫期４个阶段．

关键词：毛嵌线螺繁殖幼虫发生特征

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