吸烟大鼠淋巴细胞蛋白质组的双向电泳分析

目的应用双向凝胶电泳(Two-dimensional gel electrophoresis,2-DE)方法分析烟染毒大鼠外周血淋巴细胞蛋白质组表达的改变。方法雄性Wistar大鼠随机分正常对照组和烟染毒组,烟染毒组分小、中、大剂量,分离各组大鼠的外周血淋巴细胞,提取总蛋白,2-DE分离总蛋白,电泳凝胶银染显色,用ImageMaster 2D Platinum软件对获得的蛋白图谱加以分析,寻找差异蛋白点。结果与正常对照组大鼠图谱比较,烟染毒组图谱中与剂量有相关性变化的差异表达蛋白点有10个,其中6个随烟染毒剂量增加表达下降,4个随烟染毒剂量增加表达增强。结论2-DE可以有效分离大鼠外周血淋巴细胞蛋白质,大鼠烟染毒后淋巴细胞蛋白质的表达发生改变。

红芪黄酮气溶胶给药与灌胃给药对肺纤维化大鼠的影响

目的:比较红芪黄酮灌胃给药及红芪黄酮气溶胶给药2种给药方法对大鼠肺纤维化模型(idiopathic pulmonary fibrosis,IPF)的治疗作用。方法:选取40只SPF级雄性SD大鼠,随机分为空白组、模型组、红芪黄酮灌胃给药组及红芪黄酮气溶胶给药组,每组10只。空白组正常饮食喂养,不施加任何处理措施。其余3组大鼠使用博莱霉素气溶胶量化造模。造模后第2天开始给药,红芪黄酮灌胃给药组大鼠灌胃给药(37.41 mg/kg),红芪黄酮气溶胶给药组动物气溶胶给药(气溶胶浓度3.5~4.0 mg/m3,给药时间40 min),其余2组给予蒸馏水,每日1次,共计28天。实验结束,分别检测清醒及麻醉状态下动物肺功能;处死各组大鼠,取部分肺组织,制备匀浆,检测肺脏组织中透明质酸(hyaluronic acid,HA)和层黏连蛋白含量(laminin,LN);另取部分肺组织,分别使用HE染色及Masson染色方法,观察肺组织病理形态。结果:清醒状态及麻醉状态下造模组动物肺功能指标与空白组比较,差异具有统计学意义(P<0.05);与模型组比较,各给药组肺功能相关指标均得到显著改善,且气溶胶给药组改善更为明显。病理切片结果显示,与空白组比较,造模组出现不同程度的异常改变;与造模组比较,各给药组病理形态有显著的改善,且气溶胶给药组作用更为明显。HA、LN含量测定结果显示,与空白组比较,模型组HA、LN含量显著升高,差异均有统计学意义(P<0.05);气溶胶给药组与灌胃组比较,气溶胶给药组HA、LN含量均明显下降,差异有统计学意义(P<0.05)。结论:气溶胶给药与灌胃给药相比对肺纤维化模型大鼠治疗作用更为显著,气溶胶给药技术对于肺纤维化的治疗具有重要意义。

Transient Expression of BYDV-MP in Nicotiana benthamiana

[Objective]The aim of this study was to identify transient expression of movement protein （MP） gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus （BYDV） was cloned into potato virus X （PVX） viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.

红芪黄酮气溶胶给药与灌胃给药对COPD大鼠的影响

目的:比较红芪黄酮灌胃给药及气溶胶给药两种不同的给药方法对大鼠慢性阻塞性肺病模型(COPD)的治疗作用。方法:选取40只SPF级雄性SD大鼠,随机分为空白对照组、模型对照组、红芪黄酮灌胃给药组及红芪黄酮气溶胶给药组,每组10只。空白对照组正常饮食喂养,不加任何处理措施。其余3组每天于小动物烟雾气溶胶染毒暴露系统吸入新鲜的香烟烟雾染毒造模,每天染毒30分钟,造模28天。造模后第7天开始给药,共21天。红芪黄酮灌胃给药组每日灌胃1 mL/100 g,37.41 mg/kg,给药,红芪黄酮气溶胶给药组气溶胶给药,气溶胶浓度保持在3.5~4.0 mg/m^3,给药时间约40分钟。实验结束后,检测清醒及麻醉状态下各组动物肺功能,观察肺组织切片病理形态。结果:清醒状态及麻醉状态下造模组动物肺功能指标与空白对照组比较,均有显著性变化,差异有统计学意义(P<0.05);与模型对照组比较,各给药组显著改善肺功能相关指标,且气溶胶给药组作用更为显著。病理切片结果显示,与空白对照组比较,造模组出现不同程度的异常改变;与模型对照组比较,各给药组病理形态有显著改善,且气溶胶给药组作用更为显著。结论:气溶胶给药与灌胃给药相比,对COPD模型大鼠的治疗作用更为显著,气溶胶给药技术对于COPD的治疗具有重要意义。

Effect of TYLCV Infection on Leaf Anatomical Structure and Protective Enzyme System of Tomato

[Objective] The aim was to study the effect of tomato yellow leaf curl virus （TYLCV） infection on leaf anatomical structure and protective enzyme system of tomato. [Method] The anatomical structure of infected and healthy leaves of tomato were observed and compared by using paraffin section method. The activity changes of SOD, POD and CAT in the infected leaves of tomato were determined. [ Result] The results revealed that there were some differences in anatomical structure between healthy and infected leaves. Some cells of infected leaves were damaged so that the leaves curled and became yellow, which affected the normal function of organs. Compared with control, enzyme activities in the tomato plants infected by TYLCV were enhanced at the early periods and higher than that in control, then started to decline at the middle and late periods but lower than that in control.[ Conclusion] After infection by TYLCV, the leaf anatomical structure of tomato was changed greatly and the protective enzyme system was damaged severely, and affected the normal physJological metabolic functions of tissues and organs in tomato in further.

Impact of hepatitis C virus infection on lifestyle

AIM: To evaluate the impact of the diagnosis of hepatitis C infection on lifestyle habits such as smoking, drinking, sports activity and diet.METHODS: A self-administered, anonymous question-naire was offered to out-patients with HCV infection consecutively attending three clinical centres in Italy.RESULTS: Of the 275 respondents, 62.2% (171) were male. Mean age was 51 (range 20-80) years. Overall, after the diagnosis of hepatitis C, 74.5% of drinkers had modified (giving up or reducing) alcohol consumption, 21.3% of smokers had modifi ed their habits and 32.3% of subjects who reported sports activity had either stopped or reduced frequency of activity or chose a less fatiguing sport. Sixty-four percent of the respondents reported having modified their diet, most of them on physician's advice. CONCLUSION: After the diagnosis of hepatitis C many patients correctly modif ied their alcohol consumption and a minority their smoking habits. The reason for reported changes in diet and physical activity needs further investigations.

Viral infection of tobacco plants improves performance of Bemisia tabaci but more so for an invasive than for an indigenous biotype of the whitefly

The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus （TYLCCNV） by the invasive Q biotype and the indigenous ZHJ2 biotype of the whitefly Bemisia tabaci, a plant viral vector, as well as the influence of TYLCCNV-infection of plants on the performance of the two whitefly biotypes. Both whitefly biotypes were able to acquire viruses from infected plants and retained them in their bodies, but were unable to transmit them to either tobacco or tomato plants. However, when the Q biotype fed on tobacco plants infected with TYLCCNV, its fecundity and longevity were increased by 7- and 1-fold, respectively, compared to those of the Q biotype fed on uninfected tobacco plants. When the ZHJ2 biotype fed on virus-infected plants, its fecundity and longevity were increased by only 2- and 0.5-fold, respectively. These data show that the Q biotype acquired higher beneficial effects from TYLCCNV-infection of tobacco plants than the ZHJ2 biotype. Thus, the Q biotype whitefly may have advantages in its invasion and displacement of the indigenous ZHJ2 biotype.

Immunogenicity and virus-like particle formation of rotavirus capsid proteins produced in transgenic plants

The human pathogen, group A rotavirus, is the most prevalent cause of acute infantile and pediatric gastroenteritis worldwide, especially in developing countries. There is an urgent demand for safer, more effective and cheaper vaccines against rotavirus infection. Plant-derived antigens may provide an exclusive way to produce economical subunit vaccines. Virus-like particles, constituting viral capsid proteins without viral nucleic acids, are considered a far safer candidate compared with live attenuated viral vaccines. In this study, the rotavirus capsid proteins VP2, VP6 and VP7 were co-expressed in transgenic tobacco plants, and their expression levels, formation of rotavirus-like particles (RV VLPs) and immunogenicity were extensively studied. Quantitative real-time RT-PCR and Western blot analysis revealed that the expression level of vp6 was the highest while vp7 was expressed at the lowest levels. The RV VLPs were purified from transgenic tobacco plants and analyzed by electron microscopy and Western blot. Results indicated that the plant-derived VP2, VP6 and VP7 proteins self-assembled into 2/6 or 2/6/7 RV VLPs with a diameter of 60-80 nm. When orally delivered into mice with cholera toxin as an adjuvant, the total soluble protein extracted from transgenic tobacco plants induced rotavirus-specific antibodies comparable with those of attenuated rotavirus vaccines, while VP 2/6/7 induced higher serum IgG and fecal IgA titers compared with VP 2/6.