A/J小鼠感染猪链球菌2型后脾脏消减cDNA文库的构建及相关差异基因表达分析

为了分离与猪链球菌2型感染密切相关的基因,利用抑制性消减杂交(SSH)技术构建了8周龄A/J小鼠感染猪链球菌2型后的脾脏消减cDNA文库,并对其中169个阳性克隆进行测序,去除冗余的cDNA序列载体并聚类拼接后共获得110条差异表达序列标签(EST)。利用GenBank的BLAST软件分析比较核酸和蛋白质同源性,发现共有36个基因与这些EST具有高度的同源性,它们分别参与细胞成分、免疫、信号转导、凋亡、转录等重要功能。这些差异表达基因主要有ATP/GTP结合蛋白1(Agtpbp1)、天冬氨酸-谷氨酸-丙氨酸-天冬氨酸盒多肽5、核蛋白p68、ATP-依赖RNA解旋酶、真核生物转录因子2亚基(Eif2s2)、Na+/K+-ATP酶转运β3多肽、溶菌酶1、溶菌酶2等基因。结论:这些差异表达基因在猪链球菌2型感染过程中可能发挥重要功能。     

ALV-J诱发鸡成红细胞白血病的临床病例分析

2010—2011年检测3批病例,分别来自泰安新泰的110日龄麻鸡、济宁泗水的90日龄麻鸡和济南20日龄蛋雏鸡。病鸡均表现消瘦、精神萎靡、嗜睡、鸡冠稍苍白或发绀等,死亡率分别为20%、12%和18%。剖检,3批病鸡均可见肝脏、脾脏及肾脏严重肿大,胸腺、肌肉、腺胃等组织器官出血。新泰病鸡肝脏和肺脏表面有明显的白色肿瘤结节。血液涂片与骨髓涂片观察发现大量红细胞转变为体积较大,胞质丰富蓝染,胞核大呈圆形、核内有很纤细的染色质、核周围有空泡的成红细胞,以晚幼成红细胞为主;组织学检查发现,在所有组织脏器血管及间质内均可观察到大量聚集的与血涂片中形态相一致的成红细胞,并且可观察到核分裂相,但未见淋巴细胞聚集。通过血液学和组织学观察不仅排除马立克氏病和网状内皮组织增生症,而且排除中毒及代谢性疾病的可能。以肝脏组织DNA为模板,在3批病鸡中利用PCR检测均扩增出J亚群禽白血病病毒(ALV-J)gp85基因924bp的特异性片段,而A亚群禽白血病病毒(ALV-A)、B亚群禽白血病病毒(ALV-B)阴性。免疫组织化学检测结果表明,发病鸡肺脏和脾脏等含血量丰富的组织内发现组织细胞及成红细胞胞质呈ALV-J抗原阳性。根据以上检测结果确定此3批患有成红细胞白血病病鸡均由ALV-J感染引起。ALV-J引起单纯鸡成红细胞白血病在国内为首次发现。ALV-J在我国鸡群中的多潜能致瘤机制需要进一步的研究。     

地方品种HR土鸡不同亚型禽白血病病毒的共感染

为研究HR土鸡中存在的不同亚型禽白血病病毒(ALV)共感染的情况,本实验分别采集46只HR土公鸡的泄殖腔棉拭子和455枚鸡蛋卵白样品,采用ELISA试剂盒检测p27抗原;并采用相应的ELISA试剂盒分别检测卵黄中J亚型ALV(ALV-J)和AB亚型ALV(ALV-AB)抗体。结果表明:HR土公鸡泄殖腔棉拭子样品中p27检出阳性率为87%(40/46),卵白检出率为74.7%(340/455);而卵黄中ALV-J和ALV-AB抗体阳性率分别为0(0/30)和80%(24/30)。无菌采集初步筛选p27抗原检测为阳性的5只HR土公鸡的抗凝血接种CEF,采用抗ALV-J和ALV-A的单克隆抗体进行IFA检测,结果显示5份样品中ALV-A和ALV-J的阳性率均为100%(5/5)。同时选取HR土鸡分离株HR332进行PCR扩增鉴定,结果表明分离株HR332存在ALV-J(HR332J)和ALV-A(HR332A)。其中,HR332J与11株ALV-J国内外参考株的同源性为92.4%～97.9%;HR332A与ALV-A参考株RSA-A、MQNCSU的同源性分别为90.1%和89.7%,与国内分离株SDAU09E2的同源性为99.0%。本研究显示,地方品种HR土鸡存在不同亚型ALV共感染,同时ALV-A和ALV-J共感染同一个鸡的现象已经存在。     

西藏林芝地区核桃表型特征研究

通过对西藏林芝地区核桃表型特征进行研究,以期为西藏核桃的选育提供参考。依照核桃的性状调查方法,研究了林芝地区4个居群8个核桃家系:朗县(LX1-4)、米林县(ML7-8)、林芝县(LZ5-6)和2个铁核桃家系:波密县(BM9-10)的叶型、不同居群核桃表型特征以及坚果特征。结果表明:顶叶长与宽之比LX2最大,ML7最小;复叶长与宽比例LZ5最大,BM10最小;复叶最长的为ML7,最短者为BM10。坚果质量:BM>LX>ML>LZ;仁坚比:LX>ML>LZ>BM;壳厚:BM>LZ>ML>LX。坚果纵横径比值最大的为BM9,最小的为LX1。林芝地区朗县核桃可作为优良家系进行繁殖,波密的铁核桃保持着较高的遗传多样性,是选种和育种的优先资源。     

髓细胞瘤型J亚群禽白血病病毒四川株SCGS-1的分离鉴定及前病毒cDNA重组质粒的构建

为分析髓细胞瘤型J亚群禽白血病病毒(ALV-J)地方分离毒株的分子特征及开展该病毒的反向遗传操作研究,本试验从四川某肉种鸡场髓细胞瘤病变病鸡的组织中分离到1株髓细胞瘤型ALV-J,命名为SCGS-1.序列测定表明其前病毒cDNA全序列长7 499 bp,与其他ALV-J代表毒株序列相似性均为95％～99％.在所有基因中,其env基因和LTR序列与其他毒株相应序列同源性相对较低,env基因同源性为93.0％～99.5％,LTR基因同源性为92％～95％,pol基因同源性为97％～99％,gag基因同源性为94％～97％.根据该毒株序列和质粒pUC19的序列,将SCGS-1株前病毒cDNA全序列分3段克隆入pUC19中,构建含SCGS-1前病毒cDNA的重组载体并命名为pUC-SCGS.试验结果为研究ALV-J的进化规律和开展反向遗传操作打下基础.     

酸枣组培快繁研究

以休眠芽为试材,建立了酸枣的组培快繁体系。最适启动培养基为MS+BA1 0+IBA0 1或MS+BA2 0+NAA0 1,增殖培养基为MS+BA2 0～3 0,萌芽率和壮芽率均超过了70%。增殖培养时,下部茎段的出芽率高于上部茎段,但萌发壮芽的比率则以上部茎段相对较高。1 2MS+IBA0 5～1 0是酸枣快繁生根培养的适宜培养基,生根率最高可达92%。     

传染性喉气管炎病毒WG株糖蛋白gJ基因的克隆与表达

根据GenBank中已发表的传染性喉气管炎病毒全基因组序列（NC006233）设计引物,以传染性喉气管炎病毒WG株基因组为模板,PCR扩增糖蛋白gJ基因,并对其进行序列分析。用DNAStar软件分析酣蛋白的抗原性,选择抗原性强的140aa-483aa片段,克隆并连接到原核表达载体pET-32a（＋）中,转化BL21（DE3）菌株。经IPTG诱导后,获得大小为65．2ku的重组蛋白,命名为卜出,表达量占菌体蛋白总量的60．8％;Westernblot分析表明r-gJ具有较强的抗原性。     

CLINICAL,DIAGNOSTIC, AND PATHOLOGICAL FEATURES OF 2 CASES OF METASTATIC IRIDOPHOROMA IN A VEILED CHAMELEON (CHAMAELEO CALYPTRATUS) AND A RED-BARRED DRAGON (CTENOPHORUS VADNAPPA)

Iridophores are iridescent cutaneous pigment cells found in reptiles, fish, and amphibians. Neoplasms of iridophores are rarely reported, and little is known about their behaviour, metastatic potential, and prognostic indicators. This paper reports details of the clinical course and pathological findings of metastatic iridophoroma in a veiled chameleon (Chamaeleo calyptratus) and red-barred dragon (Ctenophorus vadnappa). The veiled chameleon presented with a subcutaneous mass on the right lateral elbow and was diagnosed as an iridophoroma on fine needle aspiration. It was otherwise clinically normal. Within 35 days of excision, multiple secondary skin masses developed which were often non-pigmented and required microscopic examination to identify them as iridophoroma metastases. There were in total 54 days between the first detection of the primary mass and end stage, extensive cutaneous and visceral metastatic disease, and there had been no detectable primary mass during routine clinical examination 128 days prior to death. The red-barred dragon presented with a white sub-mandibular mass. While undergoing an excisional biopsy the animal expired, post-mortem examination revealed a cutaneous malignant iridophoroma with tumour cell clusters seen in pulmonary arterial vessels, suggesting that surgical handling of iridophoroma could precipitate the release of circulating tumour cell clusters. This is the first iridophoroma described in a red-barred dragon and the second report of a malignant iridophoroma in a veiled chameleon. Timely detection and careful excision of lizard iridophoroma may be an important factor in clinical outcome.     

苦丁茶的组织培养研究

诱导苦丁茶具节茎段芽体发生、生长与增殖的培养基,以改良ＡｎｄｅｒｓｏｎＭＳ培养基＋ＢＡ２．０ｍｇ／Ｌ＋ＮＡＡ０．１ｍｇ／Ｌ＋ＺＴ２．０ｍｇ／Ｌ＋ＧＡ３０．５ｍｇ／Ｌ较为适宜;以改良ＡｎｄｅｒｓｏｎＭＳ培养基＋ＢＡ３．０ｍｇ／Ｌ＋ＮＡＡ０．２５ｍｇ／Ｌ＋ＺＴ１．０ｍｇ／Ｌ继代培养。培养时温度为２８℃,光照度为１５００ｌｘ,光照时间为１４ｈ／ｄ;生根处理用１／４ＭＳ＋ＩＢＡ１．０ｍｇ／Ｌ培养基。     

Jatropha tanjorensis Ellis & Saroja, a natural interspecific hybrid occurring in Tamil Nadu, India

Jatropha tanjorensis Ellis & Saroja, a species found abundantly in Tanjore, Pudukottai, Trichirapalli and Ramnad districts of Tamil Nadu state, India and grown as a fence plant showed intermediacy in phenotypic characters of J. curcas L. and J. gossypifolia L. A detailed survey at its place of occurrence supplemented with data employed from cytological and peroxidase isozyme studies revealed that J. tanjorensis is a natural interspecific hybrid between these two species. Since there was no seed set, propagation through tissue culture using leaf discs was attempted.