天然抗氧化剂Isoverbascoside诱导人胃癌MGC 803细胞分化作用研究

人胃癌 MGC80 3细胞经天然抗氧化剂 Isoverbascoside(1sov)处理后 ,细胞生长和增殖呈现Isov浓度和处理时间依赖性抑制 ,群体倍增时间也相应延长 ;2 0 μmol/ L Isov处理可使细胞形态和超微结构发生正常化变化 ,集落形成率受抑 ;胃癌分化标志酶碱性磷酸酶和乳酸脱氢酶活性均呈时间依赖性下降 ,周期表现为 G1期阻滞 .表明 Isov具有诱导 MGC 80 3细胞分化的作用 ,并可能与细胞周期阻滞有

Cloning and Expression of the Leukotoxin BSBSE Gene from Fusobacterium necrophorum

以牛源坏死梭杆菌FNn株为试材,根据GenBank上已发表的坏死梭杆菌AF312861标准菌株的lktA序列设计1对引物,利用PCR技术扩增出1 131 bp的坏死梭杆菌白细胞毒素BSBSE基因。将PCR产物插入pGEM-T Easy vector中,经双酶切鉴定正确后进行序列测定。分析表明该BSBSE序列与GenBank上已发表的坏死梭杆菌AF312861标准菌株的lktA序列的核苷酸同源性为99%,推导出的氨基酸序列同源性为98%。为研究BSBSE的免疫原性,构建了原核表达载体pMAL-p2X-BSBSE,用IPTG诱导在大肠杆菌中表达。结果表明,BSBSE基因在大肠杆菌中进行了高效特异性融合表达,融合蛋白分子量约为84.5×103,其中41.5×103为BS-BSE基因表达的蛋白质,43.0×103为MBP融合标签,Western-blotting检测表明该表达产物有免疫原性。According to the sequence of announced lktA gene in Fusobacterium necrophorum,a pair of primers were designed.The BSBSE gene was amplified by PCR.The product was cloned into pGEM-T Easy vector.When nucleotide sequence and deduced amino acid sequence were compared with homologous sequence of the FN AF312861 lktA of GenBank,the homologue of the mucleotide sequence is 99% and the homologue of the amino acid sequence is 98%.The BSBSE fragment was inserted into expression vector pMAL-p2X and the plasmid pMAL-p2X-BSBSE were expressed in E.coli BL21 by IPTG induction.The SDS-PAGE analysis indicated the weight of the fusion protein was about 84.5.0×103,which included the 41.5×103 protein expressed from BSBSE gene and 43.0×103 fusion MBP tag.The recombinant BSBSE-pMAL-p2X production has Immunogenicity with western-blotting.The cloning and expression of the BSBSE gene established the foundation of further research on the function and application of the BSBSE gene.“十五”国家科技攻关子课题(2002BA518A04);; 中国农业科学院特产研究所科研基金项目(Tcs2005-03

Method of Detecting Gastric Cancer Micrometastases in Lymph Nodes by Nest RT-PCR Amplification of Keration 19 and its Application

[目的]建立RT PCR扩增Keratin19mRNA的方法 ,评价其应用前景。[方法]采用Keratin19cDNA的套式引物建立巢式RT PCR扩增体系 ,以酶切分析及DNA点杂交法鉴定扩增产物的特异性 ,逆转录产物系列稀释法分析检测敏感性 ,并对51例临床胃周淋巴结样品作初步检测。[结果]该扩增体系具有较好的扩增特异性 ,检测敏感性达1pgRNA ,相当于从105 个淋巴细胞中检出1个胃癌细胞 ;对临床样品检测结果显示该法较病理检查法敏感性高。[结论]该扩增体系具有较好的特异性、敏感性和较高的可靠性。福建省卫生厅基金!(编号 :96075);福建省自然科学基金!(F00024)资

A Study on the mechanisms of eutrophication of a shallow upstream lake in the Jiulong River Catchment

基于对九龙江上游龙潭湖富营养化水体和沉积物现状的监测结果,通过与国内富营养化深水湖库和流域下游大型富营养化浅水湖泊进行对比,深入探讨了流域上游浅水湖泊富营养化发生的原因及主导机制.流域上游浅水湖泊具有外源污染物输入较少的特点,较下游大型浅水湖泊更易受温度等气候条件和沉积物氧化还原状态的影响,以及外源输入总磷控制具有较强的滞后效应,因此对流域上游浅水湖泊富营养化的控制必须重视内源营养盐释放,特别是结合态磷的内源释放问题.Based on site monitoring chemical data of lake water and sediments during an algal bloom in the Longtan Lake,causes and mechanisms of eutrophication in shallow upstream lakes were discussed by comparing the Longtan Lake in upstream of the Jiulong River with deep eutrophic lakes and large shallow downstream eutrophic lakes in China.The shallow upstream lake is characterized as relatively simple nutrient inputs and high susceptivity to climatic variability(such as temperature),redox conditions,and strong lag effects of the external total phosphorus inputs.Therefore,attention must be paid to the sediments nutrient releases,especially,the bound phosphorus of sediments when remediating eutrophication in shallow upstream lakes.国家自然科学基金资助项目(41175130);教育部新世纪优秀人才支持计

Cloning and Expression of the Leukotoxin Gene SH from Fusobacterium necrophorum

坏死梭杆菌白细胞毒素是坏死杆菌病的主要致病因子,白细胞毒素基因(lkT)是其编码基因。以分离到的国内牛源坏死梭杆菌fn(A)菌株f4基因组dnA为模板,应用PCr方法扩增白细胞毒素基因SH片段,克隆至PMd18-T载体上,以bAMHⅠ和HIndⅢ酶切的目的片段SH与相应酶切的PET32A载体连接构建PET32A-SH重组表达质粒,经转化E COlI bl21(dE3)后用IPTg进行蛋白诱导,SdS-PAgE检测重组蛋白表达情况。结果表明:扩增基因序列大小为1800bP,SdS-PAgE检测重组蛋白有效表达,表达得到大小为80.2kdA的目的蛋白,采用镍柱亲和层析方法纯化SH重组蛋白,获得了纯度达95%的重组蛋白;经WEST-Ern-blOT证实,该蛋白对抗坏死杆菌阳性血清具有反应活性。The leukotoxin of Fusobacterium necrophorum(FN) is considered to be one of the main virulence factors.The lkt gene encodes for FN.In this study,the SH fragment of lkt gene was amplified by PCR using the F4 genome as the template,which was isolated from the Chinese Fusobacterium necrophorum strain.The fragment was then cloned to the pMD18-T vector for sequencing.Thereafter,the SH fragment was subcloned into the multiple cloning sites of the pET32 to construct pET32a-SH recombinant plasmid,which was then trans-formed into E.coli BL21(DE3) with IPTG induction for expression.SDS-PAGE was used to analyze the recombinant protein.The results showed that the SH fragment of about 1800 bp was amplified and was about 80.2 kDa.The fusion protein was purified by Ni-NTA affinity chromatography under denature conditions,and their purity was above 95%.Western-blot analysis indicated the SH fragment had anti-genicity against Fusobacterium necrophorum.“十五”国家科技攻关子课题(2002BA518A04);吉林省科技发展计划项目(20070570);吉林市科技发展计划项目(200805

Influence of Proportion of Treg Cells and NKG2D-positive Cells by Recombinant ProTαin Mice Bearing Liver Cancer

为观察不同时期应用重组胸腺素α原(ProTα)药物干预下肝癌荷瘤小鼠的抑瘤率,并研究其对Treg细胞和NKG2D阳性细胞的影响,将36只昆明小鼠随机分成空白组、荷瘤组、药物干预组,H22小鼠肝癌细胞皮下移植建立荷瘤小鼠模型,腹腔注射重组ProTα,观察7和14d后肝癌荷瘤小鼠的抑瘤率,并检测外周单个核细胞中调节性T细胞(Treg细胞)和NKG2D阳性细胞的比例.结果表明:移植瘤7d后,药物干预组和荷瘤组瘤质量对比无显著差异;而移植瘤14d后,药物干预组瘤质量较荷瘤组有显著减小.荷瘤14d后,荷瘤组较空白组Treg细胞比例升高,NKG2D阳性细胞比例下调,差异显著;而不论是早期药物干预组还是进展期(即移植瘤7d后)药物干预组,Treg细胞比例均显著降低,NKG2D阳性细胞显著升高.由此可见,重组ProTα能够抑制肝癌荷瘤小鼠肿瘤生长,且早期、长期用药效果更好,相关作用机制涉及下调Treg细胞数量从而抑制肝癌细胞免疫逃逸,并上调NKG2D阳性细胞数量从而提高其介导的抗肿瘤效应.To observe the tumor inhibition rate by recombinant ProTαin liver cancer-bearing mice model,and to study the influence of the proportion of Treg cells and NKG2D-positive cells,36 Kunming mice were randomly divided into control group,tumor-bearing group and drug intervention group.Then a tumor-bearing mice model was established by transplanting H22 cells subcutaneously.By medicating recombinant ProTαthrough intraperitoneal injection,we observed the tumor inhibition rate after 7days and 14 days,and detected the proportion of Treg cells and NKG2D-positive cells in PBMCs.Results showed that the difference in tumor mass between the tumor-bearing group and the drug intervention group was not significant after bearing tumor for 7days.However,after bearing tumor for 14 days,the difference in tumor mass was significant.Additionally,the proportion of Treg cells increased and the number of NKG2D-positive cells declined significantly in the tumor-bearing group after bearing tumor for 14 days.No matter 14 days intraperitoneal injection of recombinant ProTαor drug intervention from the advanced stage,the proportion of Treg cells declined and the number of NKG2D-positive cells increased significantly.The results suggest that recombinant ProTαinhibits tumor growing,and the early and long-term drug intervention benefits the most.It is believed that the putative mechanism is related to that recombinant ProTαdown-regulates the proportion of Treg cells,inhibiting the immune escape of hepatocellular carcinoma cells,and up-regulates the number of NKG2D-positive cells,improving the NKG2D-mediated anti-tumor effect.福建省自然科学基金(2013J01383);; 福建省医学创新课题(2009-CXB-51

福建龙岩市龙潭湖甲藻水华成因的研究

2009年12月初,在福建龙岩市龙硿洞风景名胜区龙潭湖首次发现拟多甲藻水华,最高细胞密度达到1.11×107cells.经过应急处置,甲藻和其他藻类被杀灭.监测发现,上游小溪和溶洞地下水营养物质均优于国家Ⅱ类标准,但入湖后,TN和TP浓度明显上升.结果表明,龙潭湖蓄水改变了小溪的自然流态,形成了营养盐的累积和甲藻的快速增殖;环境和水温度的明显升高,也起到了一定的促进作用;沉积物营养盐蓄积和内释放对甲藻水华具有重要的贡献,而TP可能是本次水华的主要限制因子

白暨豚种群数量及资源保护

为了了解长江中白鱀豚的分布状况及种群数量,从1978年至1983年1月止,先后9次在长江中、下游(宜昌至南通)的干流进行了白鱀豚的生态考察,并到汉江和鄱阳湖、洞庭湖等水域查访有无白鱀豚活动的情况,依据考察所得资料,对长江白鱀豚群体的数量作了初步剖析。 通过考察,到1983年初为止,中游最上是在湖北枝城,约距长江口1,613公里的江段和下游最下是江苏太仓浏河口距长江口24公里的江段都有白鱀豚的活动。白鱀豚种群约156头,20个群体,分布在长江中、下游的17个江段里,在安庆一黑砂洲南水道约170公里及嘉鱼—

福建龙岩市龙潭湖甲藻水华成因的研究

2009年12月初,在福建龙岩市龙硿洞风景名胜区龙潭湖首次发现拟多甲藻水华,最高细胞密度达到1.11×107cells.经过应急处置,甲藻和其他藻类被杀灭.监测发现,上游小溪和溶洞地下水营养物质均优于国家Ⅱ类标准,但入湖后,TN和TP浓度明显上升.结果表明,龙潭湖蓄水改变了小溪的自然流态,形成了营养盐的累积和甲藻的快速增殖;环境和水温度的明显升高,也起到了一定的促进作用;沉积物营养盐蓄积和内释放对甲藻水华具有重要的贡献,而TP可能是本次水华的主要限制因子

固氮植物篱防治坡耕地土壤侵蚀效果研究

土壤侵蚀是坡耕地土壤退化的主要影响因子之一。在金沙江干热河谷区坡耕地的实验表明， 沿坡耕地等高线种植高密度高固氮植物篱并合理经营，可有效防治土壤侵蚀。１９９７ 年植物篱处理的土壤侵蚀量减少到对照的１４ ．５ ％ ～２３ ．９ ％ ，１９９８ 年侵蚀量仅为对照的１ ％ ～３ ％ 。坡耕地土壤侵蚀集中发生在雨季中期。土壤侵蚀过程也是土壤养分加速流失的过程，分析表明流失土壤中全氮和有机质在侵蚀土壤中含量分别是表土平均含量的２ ．３ 倍和２ ．４７ 倍以上。在培植等高植物篱系统后，经过４ ～７ ａ 的正常耕作，