Distribution of Bacteria Injected in Body of Giant Black Shrimp,Penaeus Monodon

Distribution of injected Vibrio anguillarum in body of P,,naeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue, gill, heart and haemolymph of all vibrio injected shrimp, the bacteria could be observed only 5 min after injection. The bacteria density in haemolymph, haemolymph of the hepatopancreas and gills decreased with time. In the lymphoid organ and heart, the bacteria density was the highest 48 h after injection, then decreased. Nodules could be formed in the heart, lymphoid organ and injection site.     

异育银鲫(Carassius auratus gibelio)脊髓组织细胞系的建立及对CyHV-2的敏感性

鲫造血器官坏死病是近些年来危害养殖鲫鱼最为严重的传染性疾病,其病原为鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2, CyHV-2)。本研究以异育银鲫(Carassius auratus gibelio)脊髓组织为材料,进行组织细胞体外培养,构建对鲤疱疹病毒Ⅱ型敏感的异育银鲫脊髓细胞系(Spinalcordtissuecell lines of Carassius auratus gibelio, CSC)。采用组织块移植法,进行异育银鲫脊髓细胞的原代培养,并进行传代,已传代至第128代次,获得稳定传代的细胞系后,采用低渗法进行细胞染色体核型分析,并进行鲤疱疹病毒Ⅱ型的敏感性测试与传代。结果表明,以含10%胎牛血清的L-15培养基,在24℃恒温条件下,可稳定传代培养异育银鲫脊髓细胞,传代细胞呈成纤维样或纺锤状;核型分析显示,染色体数为156±2条,该细胞为三倍体细胞。通过对CyHV-2敏感性的测试,CSC细胞感染CyHV-2后产生典型的细胞病变,感染第7天的病变细胞液的TCID₅₀达到10^9.33/mL,CyHV-2病毒可在CS...     

钻杆接头材料激光相变硬化层的组织和硬度研究

采用功率为2kW、扫描速度为100mm/min的工艺参数对钻杆接头材料30CrMnSiA钢进行了表面相变硬化处理，研究了相变层的组织和硬度特征，实验结果表明,30CrMnSiA钢表面相变硬化层分为完全淬硬层，过渡层和受热影响的基体组织，硬化层的显微组织明显细化，其表面层的硬度比高频淬火的硬度提高了30%，淬硬层深度达1.7mm，比高频淬火深度提高近1倍。     

三角帆蚌不同部位外套膜的比较研究

本文比较了三角帆蚌不同部位外套膜的细胞和组织,并用不同部位外套膜进行育珠生产试验,其结果是,由于采用不同部位外套膜细胞小片育珠、珍珠质量和产量有差异。同一横切面中外套膜三个褶比较,外褶最大,中褶最小。同一个体蚌的外褶比较,以中部外褶最大,出水管的外褶最小,同一个体内褶比较,出水管内褶最大,体后部内褶最小。从外套膜的厚薄来看,蚌的外套膜以中部及中后部较厚,中褶内外侧上皮细胞形态,细胞中高尔基体的分布和结缔组织的结构有很大的不同,说明两者发生上可能来源不同。     

文蛤外套膜组织与细胞培养

以文蛤(Meretrix meretrix Linnaeus)外套膜为材料,采用改良的M199培养基,在不同温度和盐平衡条件下进行组织与细胞培养。培养结果显示,24 h左右外套膜组织边缘有细胞爬出,48 h后开始形成单层新生细胞,并发展为生长晕,7~10 d细胞基本长满培养瓶底。细胞培养可持续30~35 d。作者还描述了培养过程中出现的3种不同形态特征的细胞,并对37℃培养条件下细胞的生长及分裂特征作了描述与讨论。     

三丁基锡(TBT)对牡蛎细胞基因组DNA影响的RAPD分析

以僧帽牡蛎(Saccuostrea cucullata)为材料,取其鳃进行组织培养,同时对其进行7个浓度0,0．02,0．2,2,20,200,2000μg／L的三丁基锡(TBT)处理,提取鳃细胞基因组DAN。从38个随机引物中筛选出4个引物,对上述DNA样品进行利用RAPD-PCR扩增分析。结果表明。TBT处理的基因组DNA与对照组样品比较有一定差异,扩增条带总数均有所减少,随TBT处理浓度的不同,条带缺失或者条带深浅变化呈明显差异性,表明不同浓度TBT处理会对DNA造成不同程度的损伤。     

太平洋牡蛎中Cg-IL-17和Cg-TGF-β的表达及其免疫作用的研究

目的:探讨Cg-IL-17和Cg-TGF-β在太平洋牡蛎中表达的组织定位、免疫作用及其可能的机制。方法:采用Real-time PCR、原位杂交和图像分析的方法进行研究。结果:实时定量PCR研究表明,Cg-IL-17和Cg-TGF-β在弧菌感染后的太平洋牡蛎的鳃、外套膜、心包液、唇须、消化道和闭壳肌中表达明显增强。原位杂交实验结果表明,Cg-IL-17和Cg-TGF-β在未染菌组牡蛎的消化道、消化腺、鳃、唇须、外套膜中都有表达;染菌组以上各组织中的Cg-IL-17和Cg-TGF-β表达量均有显著增高,而消化道、消化腺中的表达量明显高于其他组织,闭壳肌中也均有微弱表达;感染弧菌前后两种基因在性腺中均未见表达。相关性分析发现这两种基因表达量在唇须和外套膜中具有明显的相关性。结论:Cg-IL-17和Cg-TGF-β具有多种生物学效应,且都参与了太平洋牡蛎的固有免疫反应;消化道可能是牡蛎主要的免疫反应器官之一;Cg-IL-17和Cg-TGF-β在唇须和外套膜中可能存在调控关系或协同作用。     

Occurrence of Two Distinct Types of Tissue Inhibitors of Metallo-proteinases-2 in Fugu rubripes

In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 （TIMP-2） from Japanese puffer fish Fugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAs are composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteine residues, which might form six disulfide bonds as in other animals＇ TIMP-2s. Reverse-transcribed polymerase chain reaction analysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expression patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.     

大菱鲆幼鱼鳃小叶软骨组织的超微结构研究

利用电镜技术研究了大菱鲆(Scopthalmus maximus)幼鱼鳃小叶软骨组织的细胞超微结构。研究表明。鳃小叶软骨细胞呈近似的椭圆状,电子密度较低,细胞间有大量的胶原蛋白结构。软骨细胞分为3种类型：新生软骨细胞、成熟软骨细胞和衰老软骨细胞。幼稚软骨细胞体积较小,细胞核较大,具有1个核仁;胞内有丰富的粗面内质网,线粒体数量不多且体积较小;成熟软骨细胞和核的体积均庞大,具有1个核仁;可见少量的粗面内质网和线粒体;衰老软骨细胞细胞核收缩或消失。胞质中出现许多不规则空泡,细胞器很少。细胞间丰富的胶原蛋白结构使鳃小叶软骨组织具有良好的弹性。     

Molecular Cloning and Expression ofPoIR2, a Novel Gene Involved in Immune Response in Japanese Flounder （Paralichthys olivaceus）

A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed as soluble type.