A survey of seroepidemiology of toxoplasmosis

The article reports the result of serological investigation on toxoplasmosis among human,animaland fowl populations in Huimin District,Shandong Province.2269 samples from 1471 people,133pigs,343 sheeps,127 goats,75 chickens and 120 rabbits were tested by IHA method.There were 7％positives found in human,2.5～11.3％ in domestic animals and fowls.with the highest incicencein pigs.There are marked difference in incidences between the group of farmers,cadres and the group     

Acknowledgements to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and timeto the peer review,a critical process to ensure the qualityof World Journal of Gastroenterology.The editors andauthors of the articles submitted to the joumal are gratefulto the following reviewers for evaluating the articles(including those were published and those were rejectedin this issue) during the last editing period of time.     

Acknowledgments to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and time to the peer review, a critical process to ensure the quality of World Journal of Gastroenterology. The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles (including those were published and those were rejected in this issue) during the last editing period of time.     

Acknowledgements to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and timeto the peer review,a critical process to ensure the qualityof World Journal of Gastroenterology.The editors andauthors of the articles submitted to the journal are gratefulto the following reviewers for evaluating the articles(including those were published and those were rejectedin this issue) during the last editing period of time.     

Acknowledgments to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and time to the peer review,a critical process to ensure the quality of World Journal of Gastroenterology. The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles (including those published in this issue and those rejected for this issue) during the last editing time period.     

Acknowledgments to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and time to the peer review, a critical process to ensure the quality of World Journal of Gastroenterology. The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles (including those were published and those were rejected in this issue) during the last editing period of time.     

Acknowledgements to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and time to the peer review, a critical process to ensure the quality of World Journal of Gastroenterology. The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles (including those were published and those were rejected in this issue) during the last editing period of time.     

Acknowledgments to Reviewers of World Journal of Gastroenterology

Many reviewers have contributed their expertise and time to the peer review, a critical process to ensure the quality of World Journal of Gastroenterology. The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles (including those were published and those were rejected in this issue) during the last editing period of time.     

Establishment of transgenic mice carrying the gene of human nuclear receptor NRSA2 (hB1F)

AIM: Human hepatitis B virus enhancer Ⅱ B1 binding factor (hB1F) was cloned and characterized as a novel member of the Ftz-F1 (NRSA) nuclear receptor subfamily. Although progresses have recently been made, its biological function remains largely unidentified. The aim of this study was to establish an hB1F transgenic mouse model to promote the functional study of hB1F. METHODS: Transgene fragments were microinjected into fertilized eggs of mice. The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offsprings were identified by PCR and Southern blot analysis. Transgene expression was analyzed with RT-PCR and Western blot analysis. Transgenic founder mice were used to establish transgenic mouse lineages. The F1 and F2mice were identified by PCR analysis. RESULTS: Seven mice were identified as carrying copies of transgene. RT-PCR and Western blotting results showed that the transgene was expressed in heart, liver, lung, kidney and stomach in one of the transgenic mouse lineages.Genetic analysis of the transgenic mice demonstrated that the transgene was integrated into the chromosome at a single site, and was transmitted stably. CONCLUSION: In this study we established an hB1F transgenic mouse model, which will facilitate the investigation of the biological function of hB1F in vivo.     

Gene expression profile in liver of hB1F transgenic mice

AIM: To analyze the tissue morphologic phenotype and liver gene expression profile of hB1F transgenic mice. METHODS: Transgene expression was analyzed with RT-PCR and Western blotting. For one of the transgenic mouse lines, tissue expression pattern of the transgene was also examined with immunochemical methods. Pathological analysis was used to examine the tissue morphologic phenotype of established transgenic mice. The liver gene expression profile of transgenic mice was analyzed with microchip, and some of the differentially expressed genes were verified with RT-PCR. RESULTS: The expressions of hB1F were shown in livers from 6 of 7 transgenic mouse lines. The overexpression of hB1F transgene did not cause pathological changes. Expressions of three genes were up-regulated, while down-regulation was observed for 25 genes. CONCLUSION: The overexpression of hB1F transgene may cause changes of gene expression profiles in the liver of transgenic mice.     

Generation and characterization of a transgenic mouse model for pancreatic cancer

AIM: To generate a SV40Tag transgenic tumor animal model and to study the mechanism underlying tumorigenesis. METHODS: A mammary gland expression vector containing SV40Tag DNA was generated. Transgene fragments were microinjeted into fertilized eggs of FVB mice. The genetically manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice. PCR and Northern blot analysis were used for genotype analysis of F1 and F2 mice. Transgene expression was detected by RT-PCR and immunohistochemistry. RESULTS: SV40Tag gene was detected in two lines of transgenic mice. One of them delivered the transgene to Fl and a tumor was found in the pancreas of these mice. RT-PCR and immunohistochemistry showed that SV40Tag gene was expressed in the tumor. Pathological characterization of the transgenic mice demonstrated that the tumor belonged to pancreatic cystic neoplasm. CONCLUSION: SV40Tag transgenic mouse model can be successfully established. The transgenic mice develop a pancreatic tumor, which can be used for investigation of the molecular mechanism of tumorigenesis in vivo.     

早老性痴呆转基因小鼠的研究

目的 建立早老性痴呆的转基因小鼠,为进一步的发病机理研究及药物筛选提供动物模型。方法 显微注射方法制备转基因小鼠,通过ＰＣＲ、Ｓｏｕｔｈｅｒｎ杂交鉴定。结果 质粒ｐＰｄＡＰ７５１的Ｔｔｈ１１１Ⅰ＋ＸｂａⅠ双酶切片段长度为４．３ｋｂ,经回收、纯化后,注射到小鼠受精卵的雄性原核,经ＰＣＲ,Ｓｏｕｔｈｅｒｎ杂交鉴定,确定首建鼠一只。首建鼠现已传代建系。结论 通过显微注射的方法,建立了早老性痴呆的转基因动     

肝癌相关基因DLK1转基因小鼠的构建与鉴定

目的 构建并鉴定肝脏特异性表达的DLKl转基因小鼠.方法 通过基因重组方法,将小鼠DLK1 cDNA片段置于小鼠白蛋白基因增强子和启动子序列下游,构建肝脏特异性表达的DLKl重组质粒,酶切重组质粒得到转基因片段,转基因在体外进行表达鉴定后,显微注射获得DLK1转基因首建小鼠,对首建小鼠进行传代,利用F1代小鼠进行DLK1转基因表达的鉴定.结果 逆转录(RT)-PCR和细胞免疫荧光显示,转基因DLK1片段可在小鼠肝癌细胞系Hep1-6中表达.RT-PCR和免疫组化结果显示,DLK1在F1代成年转基因小鼠肝脏中特异性表达.结论 成功构建了肝脏特异性表达的DLK1转基因小鼠.     

肝脏特异性表达人遗传印记基因PEG10转基因载体的构建和鉴定

目的构建小鼠肝脏特异性表达人遗传印记基因PEG10的转基因载体pALB—PEG10-EGFP，为制备转基因小鼠做准备。方法RT—PCR扩增PEG10基因cDNA序列，克隆入T载体进行酶切、测序鉴定，后将其定向克隆至真核表达载体pALB．EGFP中ALB的下游，构建转基因载体pALB—PEG10-EGFP；在Lipofectamine介导下转染L02细胞，经G418抗性筛选，挑选阳性克隆并扩大培养；采用RT—PCR、Westernb1ot、免疫细胞化学等方法分析PEG10在L02细胞中的表达和细胞内定位。结果酶切和测序结果表明pALB—PEG10-EGFP构建成功；稳定转染后的L02表达有PEG10的mRNA及蛋白，且主要定位于胞浆。结论重组体pALB—PEG10-EGFP的构建初步奠定了转基因小鼠制备的基础。     

Replication and gene expression of mutant hepatitis B virus in a transgenic mouse containing the complete viral genome with mutant s gene

AIM: To establish the transgenic mouse line harbouring complete hepatitis B virus (HBV) genome with mutant s gene (adr subtype). METHODS: Transgenic mice were generated by microinjecting HBV genome into fertilized eggs. Integration, expression, replication of HBV gene and histological changes in transgenic mice were estimated by genomic DNA PCR, serum DNA PCR, Southern blot, ELISA, HE staining, immunohistochemistry and transmission electron microscopy. Transgenic mice with HBsAg positive in serum were bred and analyzed. RESULTS: A total of 288 eggs survived from microinjections were transplanted into the oviducts of 13 pseudopregnant mice and 49 pups were produced. Twenty-six mice were identified to have the integrated HBV gene. Serum HBsAg and HBeAg were detected in 2 of 43 mice. HBsAg and HBcAg in cytoplasm or nuclei of hepatocytes were detected in 10 mice. Founders with HBsAg in serum were named lineages G145R-15 and G145R-18. Of the 16 F1 offsprings generated by G145R-15 founder, 12 were positive for HBV genome with PCR, 10 were positive for HBsAg and HBcAg with immunohistochemistry and 7 were positive for HBsAg and HBeAg with ELISA. Only 1 of 8 F1 offsprings generated by G145R-18 founder was survived and it was detected positive for HBV genome, HBsAg, HBcAg and HBeAg. Both of the two lineages had some pathological characteristics of mild chronic hepatitis B in the liver, such as swelling of hepatocytes and focal hepatocellular necrosis and parenchymal lymphomononuclear cell infiltrate. CONCLUSION: Transgenic mice harbouring HBV with mutant s gene can be generated. The HBV genes are integrated in the transgenic mice genome and can be expressed, replicated, packaged and excreted. HBV DNA can be stably transmitted in the transgenic mice.