Peripheral neuropathies, hepatocellular carcinomas and islet cell adenomas in transgenic mice

The ability to introduce foreign DNA into the genome of mice offers unique opportunities to produce new models of disease process. Recent experiments have shown that integration and expression of simian virus 40 (SV40) T antigen genes and the murine mammary tumour virus (MMTV)-myc genes in transgenic mice can lead to the development of neoplasia in a remarkably tissue-specific manner. In the case of SV40-bearing mice, tumours consistently develop in the choroid plexus. In the accompanying paper, we show that the 72-base pair (bp) enhancer in the SV40 genome is instrumental in directing tumorigenesis to the choroid plexus. However, when the enhancer is deleted from a construction also containing the metallothionein-human growth hormone fusion gene (SV delta e-MGH), an entirely new pattern of pathology results. The present report focuses on transgenic mice carrying this construct; they develop demyelinating peripheral neuropathies, hepatocellular carcinomas and islet cell adenomas.     

Tumour necrosis factor alpha restores granulomas and induces parasite egg-laying in schistosome-infected SCID mice.

Schistosomiasis (bilharzia) is a parasitic disease caused by several species of schistosome worms (blood flukes). The key pathogenic event in this disease is the formation of granulomas around schistosome eggs trapped in portal venules of the liver. Granulomas are a distinctive form of chronic inflammation characterized by localized aggregation of activated macrophages around an inciting stimulus. Each granuloma evolves to form a fibrous scar; in schistosomiasis, the result is widespread hepatic fibrosis and portal hypertension. To identify the specific immune signal molecules necessary for granuloma formation, we studied schistosome infections in severe combined immunodeficient (SCID) mice, which have normal macrophages but lack functional B or T lymphocytes. Here we report that the immunoregulatory cytokine tumour necrosis factor alpha is necessary and sufficient to reconstitute granuloma formation in schistosome-infected SCID mice. Moreover, we find that the parasitic worms require tumour necrosis factor alpha for egg-laying and for excretion of eggs from the host. The implication of this latter result is that the parasite has adapted so successfully to its host that it uses a host-derived immunoregulatory protein as a signal for replication and transmission.     

Efficient transfer and expression of human clotting factor IX cDNA in neonatal hemophilia B mice mediated by VSV-G pseudotyped retrovirus

The feasibility of in vivo gene therapy for hemophilia B by VSV-G pseudotyped retroviral vector was introduced. The novel packaging cell line 293GPG was used to produce VSV-G/G1NaBAIX pseudotyped virus with the highest titers up to 8.5×10⁸ cfu·mL^-1. In contrast to the conventional retrovirus, VSV-G pseudotyped virus was more resistant to inactivation by serum complements (P＜0.001). Our results also demonstrated that VSV-G pseudotyped virus was more stable in neonatal mice serum than in adult mice serum (P＜0.01). After intraperitoneal injection of different doses of virus, hFIX antigen was detected and lasted for more than 120 d, the highest level reached (72.5±6.1) ng·mL^-1. Moreover, the functional activity was improved to some extent in all hFIX-treated mice, the most remarkable improvement was observed in the mice treated with higher dose of virus whose clotting activity increased to (3.4±1.5)% and APTT (activated partial thromboplastin time) reduced to (43.2±7.2) s. The anti-hFIX antibody was not detected by the method of Bethesda, no germ line transmission and any side effects associated with gene transfer were found. Our results indicated that neonatal gene therapy for hemophilia B mice by VSV-G pseudotyped retrovirus is promising.     

Expression of endogenous murine leukaemia viruses in AKR/J streaker mice

MICE of the AKR/J strain show high rates of spontaneous thymic lymphoma and are chronically infected with murine leukaemia viruses. Total thymectomy in young mice of this strain results in a marked decrease in the incidence of leukaemia. The finding of athymic mutant mice on the AKR/J background (referred to as streaker mice) is of importance in the study of genetic factors which influence virus expression and leukaemogenesis. Here we report that mice of both normal and mutant genotypes vary in the expression of a particular class of murine leukaemia virus. This lower virus expression in conjunction with the absence of a thymus leads to a lower tumour incidence in streaker mice.     

猪生长激素基因与白介素-6基因在小鼠体内的表达效应分析

比较研究了阳离子脂质体包裹VpGH、VpGH联合VpIL-6以及脂质体(对照)腹腔注射昆明小鼠的增长效应与免疫应答.结果表明, VpGH注射小鼠、VpGH联合VpIL-6注射小鼠在体重增长幅度和血清GH水平上均较对照组显著增高,在注射后第4周效果最明显;VpGH联合VpIL-6注射小鼠较仅注射VpGH小鼠在增重幅度和血清GH水平显著增高的趋势明显.但在白细胞数量、红细胞数量上较对照组除第1,2两周外,其余时间段无显著差异.在IgG表达上各组均无显著差异.     

枸杞多糖对Ⅱ型糖尿病小鼠胰岛细胞形态与功能的影响

采用高糖高热量饮食并结合小剂量腹腔注射STZ诱导Ⅱ型糖尿病（NIDDM）小鼠模型,枸杞多糖灌胃4周,用免疫组化技术对胰岛A、B细胞进行形态学分析及观察血糖、血清胰岛素的变化。结果显示,枸杞多糖大,小剂量组和模型组比较空腹血糖下降,胰岛素水平升高。图像分析大剂量组和模型组比较,B细胞数密度增加、A细胞数密度减小、B细胞切面积、核面积、核质比升高。结论：枸杞多糖可以改善Ⅱ型糖尿病小鼠胰岛细胞形态和功能并促进胰岛素的分泌,有良好的降血糖作用。     

内源激活剂驱油藏内微生物菌群结构变化分析

油藏内源功能微生物的激活能有效提高采油效率。其中激活后功能微生物的群落变化分析及相关代谢基因丰度的变化对内源激活剂的筛选及激活机理研究具有重要的指导意义。本实验利用高通量测序技术准确分析了内源激活剂注入井（WJW199.1）和对照井（WJW202）中内源微生物（包括古菌和细菌）基于门、纲、属水平的种群多样性和群落结构。结果表明采出水中Thaumarchaeota门的古菌占主导,而丰度最高的细菌属于Proteobacteria门和 Firmicutes门。不管对于古菌还是细菌,注入井中的微生物多样性都远高于对照井,说明激活剂的添加有效激活了其中的内源微生物,提高了群落结构多样性。尤其是Pseudomonas,Arcobacter等具有产表活剂能力的功能微生物。同时,硫酸盐还原菌脱硫弧菌属等有害菌被抑制,丰度降低。表明从微生物种群变化的程度来看内源激活剂的注入降低了有害菌的丰度,激活了功能微生物,增强了整体微生物的驱油能力。     

紫菜腐霉侵染条斑紫菜叶状体过程研究

对我国北方沿海条斑紫菜栽培海区发生的条斑紫菜赤腐病进行了研究.揭示了病原菌紫菜腐霉(Pythium porphyrae)侵袭条斑紫菜叶状体的过程.紫菜腐霉的丝状菌丝侵袭并穿透紫菜细胞,导致细胞色素溶出,细胞萎缩,最后死亡.紫菜腐霉发育到一定阶段,在菌丝末端形成孢子囊,孢子囊成熟后,形成排放管,放出游孢子,游孢子可以再次侵袭紫菜细胞.在一定条件下,紫菜腐霉可以进行有性生殖,通过藏精器与藏卵器结合,形成卵孢子,卵孢子沉入海底,在条件适宜时可再次萌发.冷藏网上细胞同样可以被病原菌所感染,然而使用冷藏网可以阻止病害的蔓延,并能提高紫菜品质.     

白藜芦醇诱导VEGF表达促进缺血再灌注小鼠新生血管形成

研究白藜芦醇能否诱导小鼠缺血,再灌注后缺血区域新生血管形成及其与血管内皮细胞生长因子(VEGF)表达的相关性,用90只雄性BALB/c小鼠随机分为单纯缺血组、白藜芦醇组及假手术组,采用大脑中动脉线栓法制作局灶性脑缺血模型,免疫组化法测定微血管密度,Western blot检测VEGF蛋白表达.结果白藜芦醇处理组与单纯缺血对照组微血管计数存在差异,并且有统计学意义(P<0. 05),白藜芦醇处理组在各时间点VEGF蛋白表达水平均较高(P<0.05).说明白藜芦醇可以诱导脑缺血后VEGF早期表达,促进缺血区域新生血管形成,发挥脑保护作用.     

油茶种子性状及浸种后内源激素含量分析

【目的】研究油茶种子性状,分析油茶种子浸种期间吸水率及吸水速率,探讨种子浸水机制下内源激素动态变化规律,为标准化砧木培育提供技术支撑。【方法】以‘长林3号’(3号)、‘长林4号’(4号)、‘长林40号’(40号)、‘长林18号’(18号)、‘长林53号’(53号)种子为试验材料,测定种子形状参数、体积,并在人工气候控制条件下计算不同浸种时间的吸水率和吸水速率;利用液质联用(LC-MS)方法对‘长林18号’(18号)种子浸种不同时间和温度条件下赤霉素(GA₃)、脱落酸(ABA)、水杨酸(SA)和茉莉酸(JA)、生长素(IAA)和玉米素核苷(ZR)含量进行检验,采用单因素方差分析比较不同条件下各激素含量差异。【结果】品种间种子体积由大到小依次为长林53号、40号、18号、3号、4号,且53号与其他品种存在显著差异;种子三维系数显示长林3号、4号和40号多呈扁平状,18号和53号多近球形,且种子三维系数与种子横径长度显著相关;3号和4号种子体积与纵径存在显著或极显著正相关,40号和53号横径与侧径存在显著负相关,18号种子体积与形状参数无相关性。各品种浸种48 h后吸水率和吸水速率逐渐稳定;种子浸种0~21 h,53号吸水率最高,40号最低;浸种21~48 h吸水率由大到小依次为4号、3号、53号、40号和18号。各品种吸水速率在浸种10 h内,由大到小依次为40号、4号、3号、53号、18号,浸种10 h后53号最高。种子浸种后的发芽率无显著差异,由大到小依次为4号、40号、53号、3号和18号。18号浸种后GA₃含量逐渐上升,在浸种2 d时达到最高,为0.39 mg/kg;SA和JA含量在浸种4 d时达到最高,分别为0.058 mg/kg和1.77 mg/kg;ABA含量在浸种2 d和5 d时达到最高,约为0.050 mg/kg,4 d时最低为0.014 mg/kg。m(GA₃)/m(ABA)在浸种1 d和4 d时维持较高水平。m(GA₃)/m(JA)在浸种1 d时达高值,4 d时最低;m(GA₃+SA)/m(ABA+JA)随着浸种时间增加逐渐下降。18号浸种温度为25~30 ℃时对提高种子内源激素含量有着显著作用。【结论】横径显著影响种子形状,18号种子均匀度高且吸水率和吸水速率较为稳定;种子浸种后48 h吸水率和吸水速率达到稳定状态,促进萌发类激素含量升高。因此,浸种温度应在30 ℃下有利于种子萌发。研究结果可为油茶砧木标准化培育提供理论依据。     

Transduction of endogenous envelope genes by feline leukaemia virus in vitro

Feline leukaemia viruses (FeLV) are exogenous retroviruses that can be detected in most cats with leukaemia, aplastic anaemia, myeloproliferative diseases and fatal immunosuppression. FeLV isolates have been divided into three subgroups, based on the viral envelope-determined properties of interference and host range in vitro. FeLV-A is present in all natural isolates and is generally minimally pathogenic. FeLV-B is found with FeLV-A in isolates from approximately 40% of natural infections and in a higher percentage of cats with lymphoma. Following the fundamental observations of genetic reassortment of avian retroviruses with endogenous viral genes and the origination of lymphomagenic viruses during the ontogeny of AKR mice, we show here that transfection of feline cells with FeLV-A DNA results in its recombination with endogenous FeLV-related sequences to produce viruses with the structural and host range properties of FeLV-B. Thus in vitro propagation of a retrovirus may result in the generation of variants with very different properties.     

HLA class II induction in human islet cells by interferon-gamma plus tumour necrosis factor or lymphotoxin

HLA class II molecules are surface glycoproteins which are essential in the initiation of immune responses. It has been postulated that induction of class II in epithelial cells such as endocrine cells, which are normally class II negative, may result in autoimmunity. In type I diabetes, islet beta cells, the target of the autoimmune process, selectively express class II antigens. But in contrast to most other cell types, islet beta cells are not stimulated to express class II by interferon-gamma (IFN-gamma) and thus the conditions under which this induction occurs have been particularly elusive. The cytotoxins tumour necrosis factor (TNF) and lymphotoxin (LT) synergize with IFN-gamma in a number of activities. We report here that IFN-gamma in combination with either TNF or LT induces islet cell class II expression. This finding has important implications for the pathogenesis of type I diabetes and the understanding of the differential control of class II expression.