鲤和草鱼杂交四倍体及其回交三倍体草鱼杂种的研究

进行了兴国红鲤♀×草鱼♂的杂交,杂种一代染色体数为142—156,众数值为146—152,系染色体自动加倍的异源四倍体。其外形和习性接近母本,两性均能成熟,雌、雄鱼分别与草鱼进行了回交。以草鱼为母本,杂种一代为父本的回交子代全部为草鱼形,草食性。以杂种一代为母本,草鱼为父本的回交子代一部分象草鱼,一部分象鲤,但草鱼形子代成活率较低,两个回交组合的子代均为三倍体,染色体数98—100,众数值98。1983年获得的杂种一代♀×草鱼♂的鲤鱼形回交杂种,已于1986年性成熟,雌、雄鱼分别与草鱼再次回交,获得了二次回交杂种。测定了红鲤、草鱼、杂种一代及其与草鱼回交子代的乳酸脱氢酶和过氧化物酶的同工酶,它们各有不同的电泳图谱。     

草鱼与团头鲂肠道菌群结构比较分析

选取湖泊养殖的草鱼（Ctenopharyngodon idellus）和团头鲂（Megalobrama amblycephala）为研究对象, 通过对其消化道细菌群落16S rDNA进行细菌群落聚合酶链式反应-变性梯度凝胶电泳（PCR-DGGE）分析, 比较了其消化道微生物群落结构。在两种草食性鱼类消化道中分别检测到不同的谱带, 其中草鱼的平均谱带数为（33.32.8）条, 团头鲂的平均谱带数为（38.02.5）条。基于所得PCR-DGGE指纹图谱谱带丰度值数据的UPGMA聚类分析和PCA排序均显示草鱼消化道微生物群落并没有与团头鲂消化道微生物群落分开; 相比于草鱼和团头鲂消化道微生物群落之间的差异, 草鱼和团头鲂个体间差异更加明显, 而且草鱼消化道微生物群落的个体分化更为明显。对特定条带的切胶回收测序结果显示, 草鱼和团头鲂消化道内检测到的菌群都主要来自-变形菌门、梭杆菌门和厚壁菌门, 另外草鱼消化道中还检出少量拟杆菌门细菌。以上结果表明同一生境中食性相同的野生草鱼与团头鲂消化道微生物群落组成不存在显著差异, 条带回收测序检测到相似的微生物种类。研究结果补充了人们在食性对消化道微生物群落结构影响方面的认识, 同时也暗示草鱼和团头鲂消化道内微生物群落对营养物质的代谢方式类似, 这为深入研究草食性鱼类消化道微生物菌群参与营养物质代谢积累了资料。       

草鱼外周血细胞数量及其在胁迫或感染下的变化

为了提供可反映草鱼(Ctenopharyngodonidellus)健康状况的血细胞指标的基础数据,研究了4种质量规格组的正常草鱼的外周单位体积血细胞数量,以及分别在硫酸铜或敌百虫处理下和多子小瓜虫感染后的变化。结果表明,1000g组草鱼的单位体积红细胞数显著高于其他3种质量规格组;60g组草鱼的单位体积白细胞数显著低于其他3种质量规格组。硫酸铜或敌百虫处理组的单位体积红、白细胞数显著高于对照组;多子小瓜虫感染组的单位体积白细胞数极显著高于对照组。这里提出一个初步参考指标,60～1000g健康草鱼的红细胞值为1.15×10⁶～3.33×10⁶cells/μL,白细胞值为2.20×10⁵～4.40×10⁵cells/μL。     

二龄鲢和鳙血液的比较研究

以１１５尾二龄链和９１尾二龄鳙为材料,测定了血液中红细胞数,白细胞数、血红蛋白值、红细胞脆性植、比积及比重,红细胞沉降率,白细胞分数计数值及各型血细胞大小等正常值。结果显示：红细胞和白细胞值,比积值及淋巴细胞所占百分比值均是鲢高于鳙,而红细胞沉降率和中性粒细胞却是鳙高于链,在鲢上,其血液指标植也存在雌雄差异,上述血液指标值也存在周年变化。     

革胡子鲶血液常数值的周年变化

在淡水养殖鱼类血液学研究中,有关革胡子鲶血液常数值及其周年变化的资料尚属罕见,而这方面的研究对了解革胡子鲶生物学性状和促进其繁殖均有一定意义。为此,作者以62尾(雌鱼32尾,雄鱼30尾)健康无病、体长均值为 31.55±4.92 厘米、体重均值为 281.33±145.64克、年龄为1—2 周龄的革胡子鲶(Clarias lazera)血液为材料,对其红细胞数(RBC)、白细胞数(WBC)、血红蛋白值(Hb)、比积(Ht)、比重(BSG)、红细胞脆性(Eof)、沉降率(ESR)、白细胞百分式(DLC)和各型血细胞的大小(长径×短径)等常数值分双月进行了周年测定,并分析了性别、繁殖等因素对上述常数值的影响和周年变化的规律。     

GCRV感染存活草鱼的血清蛋白表达特性

为挖掘草鱼关键抗性分子,研究利用4D label-free定量蛋白质组学技术系统分析了团队前期获得的GCRV感染存活草鱼(候选抗性草鱼)与对照草鱼的血清蛋白表达差异。共鉴定到858个草鱼血清蛋白, 329个蛋白在两类草鱼中差异表达,其中163个蛋白在候选抗性草鱼血清中显著高表达, 166个蛋白显著低表达。差异表达蛋白的功能主要注释为体液免疫反应调节相关蛋白,显著富集到补体凝血级联、细胞黏附和铁死亡等信号通路。对差异表达的免疫相关蛋白进行分析发现,候选抗性草鱼血清中体液免疫分子MASP2、C4a、C4b、C7b、C8b、C8g、C9、CFI、C3a.1、C3a.2、C3a.3和C3a.6等补体和抗原提呈相关免疫分子MHC1UBA和IGL4V8等蛋白表达水平显著高于对照草鱼,而T细胞免疫相关分子TRAV、IGHV1-1、IGHV2-1、IGHV6-1、IGHV3-2和IGHV11-2等蛋白表达水平显著低于对照草鱼。免疫印迹检测进一步证实,以C3和IgM为代表的体液免疫分子在候选抗性草鱼血清中显著高表达,可能与草鱼抗病能力关联。研究将为高抗性草鱼的选育提供可参考的分子资源。     

雌核发育抗病草鱼与普通草鱼的免疫学指标比较

雌核发育抗病草鱼在养殖过程中已表现出较好的抗病能力,但对其抗病力的确定还缺乏有力的证据。通过对雌核发育抗病草鱼和相同饲养条件下普通草鱼的主要免疫球蛋白基因(Ig D、Ig Z和Ig M)相对表达量、免疫生化指标(白蛋白、碱性磷酸酶、谷丙转氨酶、谷草转氨酶和肌酐)、血清溶菌酶活性等相关参数的比较,分析了雌核发育抗病草鱼的免疫能力。数据显示雌核发育抗病草鱼Ig Z、Ig M m RNA在血液和脾脏中的相对表达量、Ig D m RNA在头肾和脾脏中的相对表达量都与普通草鱼存在显著差异(P0.05);溶菌酶活性与普通草鱼存在显著差异(P0.05)。在免疫生化指标方面,雌核发育抗病草鱼也明显优于普通草鱼。研究结果肯定了雌核发育抗病草鱼在免疫机能方面的优越性。     

三种淡水鱼类在中国南北两个地区的肠道菌群差异比较

山东省德州市禹城市和湖南省长沙市望城区分别属于我国北方和南方地区,环境和气候差异明显。本试验以禹城市和望城区两地的草鱼(Ctenopharyngodon idella)、鲫鱼(Carassius auratus)和鲤鱼(Cyprinus carpio)为研究对象,对肠道中的菌群进行16s rRNA V4高变区扩增,基于IonS5 TM XL测序平台进行测序,并对相关数据进行分析。结果表明,在门水平上鲫鱼、鲤鱼、草鱼的肠道优势菌主要为变形菌门(Proteobacteria)、梭杆菌门(Fusobacteria)、硬壁菌门(Firmicutes),拟杆菌门(Bacteroidetes),各组中这四种菌门所占比之和均在80%以上;基于Bray-Curtis距离值的秩次进行组间差异显著性检验,山东草鱼组和湖南草鱼组比较中组间差异大于组内差异(R=0.307,P<0.05),且线性判别分析显示湖南草鱼组的特征性菌群为弧菌属,而山东草鱼组的为梭菌纲,推测特征性菌群差异与环境的影响有关;有害菌种的统计分析表明,湖南鲫鱼组和鲤鱼组的嗜水气单胞菌属相对丰度明显高于对应的山东鲫鱼组和鲤鱼组,山东草鱼组和鲤鱼组的弧菌属相对丰度高于对应的湖南草鱼组和鲤鱼组,可能受外界环境的影响,不同地区抵御外来有害细菌的能力也不一样。本研究对两地的淡水鱼类肠道微生物群落结构进行了比较和分析,为进一步的鱼类发育研究奠定基础。     

草鱼中国土著群体与欧美日移居群体遗传差异的线粒体序列分析

草鱼是中国的土著鱼类,自20世纪60年代以来已被移居到100多个国家和地区,主要用来控制水草或水产养殖。本研究通过线粒体D-Loop区（764bp)和COII＋tRNA基因(719bp)序列分析,了解草鱼的中国土著群体（长江、珠江、黑龙江）和国外移居群体（匈牙利多瑙河、美国密西西比河、日本利根川河）之间以及各地方群体间的遗传差异。结果表明,中国土著草鱼群体的遗传变异高于国外移居群体;分子方差分析（AMOVA）表明,不同草鱼群体的遗传变异主要来自群体内,而不同地域间的差异极少;群体两两间Fst值比较表明,大多数群体之间遗传差异极显著;由TCS构建的单倍型网络结构图显示,长江草鱼群体是最原始的群体,其他水系草鱼均由长江群体演化而来;通过基因流分析发现,匈牙利多瑙河群体和日本利根川河群体来自长江和黑龙江群体,美国密西西比河群体除引自长江、黑龙江水系外,还有部分引自于珠江水系。     

草鱼野生和养殖群体间遗传变异的微卫星分析

运用微卫星标记对江苏境内草鱼(Ctenopharyngodonidella)一个野生群体(邗江群体)和两个养殖群体(淡水中心群体和无锡前洲群体)遗传多样性进行了分析。在10个座位中,每个座位检测到的等位基因数2~8个。有效等位基因数、多态信息含量、期望杂合度、平均表观杂合度均以邗江草鱼野生群体最高,分别为3·9、0·5068、0·6939、0·7;无锡前洲草鱼养殖群体最低,分别为2·2、0·1796、0·5235、0·5286;淡水中心草鱼养殖群体各参数均介于两者之间,分别为3·5、0·2902、0·5418、0·5429。以上结果表明:草鱼野生群体遗传多样性更为丰富,而草鱼养殖群体存在杂合度降低,遗传多样性下降的现象。邗江草鱼野生群体与淡水中心草鱼养殖群体和无锡前洲草鱼养殖群体间遗传分化系数分别为0·219和0·246,而两个草鱼养殖群体间遗传分化系数为0·034。这表明草鱼野生群体与草鱼养殖群体间分化严重,而草鱼养殖群体间分化微弱。各座位分化程度的χ2检验结果表明,10个座位中有GM18、MFW1-1、MFW1-2三个座位群体间分化达到极显著水平,GM03-2、MFW5两个座位群体间分化差异显著,其他座位分化不显著。针对每个座位对各群体进行Hardy-Weinberg平衡检验发现:由于草鱼养殖群体在GM03-1、GM03-2、GM18三个位点杂合子缺失,草鱼野生群体在位点GM19杂合子过剩而严重偏离平衡。实验表明:近交容易引起草鱼遗传多样性下降,纯合速度加快。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.