1.Progress in Research of the Role of Hepatitis C Virus Protein in Cell Signal Transduction Pathway
Mei XUE ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2009;17(6):470-474
Abnormal cell signal transduction is associated with the occurrence and development of human diseases. Some virus pathogenicity and infection mechanism are due to virus antigen protein acting on the host cell signal transduction pathway, leading to host cell signal transduction disorder. Hepatitis C virus (HCV) is a major pathogen of chronic hepatitis C, which causes cirrhosis and hepatocellular carcinoma. But the pathogenesis of HCV and persistent infection mechanism remain far from clear. HCV pathogenesis may be related to the HCV protein expression interfering host cell signal transduction pathways. The studies of hepatitis C virus proteins acting on host cell signal transduction pathways, not only help to clarify the impact of its pathogenic mechanisms, but also benefit to new drug design and development for new treatment methods. This article summarizes the recent progress in research on the effect of hepatitis C virus protein in cell signal transduction pathways in the past few years.
2.Dialectical Thought of Interdisciplinary——Training of Cross-talent in Life Sciences
Gexin XIAO ; Lijuan FU ; Jiejie DAI
Chinese Journal of Medical Education Research 2002;0(01):-
The history of life sciences has displayed that Interdisciplinary contributes to the development of life sciences.It also causes people to think dialectically,what is regarded as the development of Interdisciplinary,what is its drive causes,what is the rule of its development.By exploring the developing trend of scientific and interdisciplinary science,we have fully discussed how to train the Cross-talents.
3.Establishment of a tree shrew model of Fusarium keratitis
Yan LI ; Jiejie DAI ; Xiaomei SUN ; Hai LIU ; Zhulin HU
Acta Laboratorium Animalis Scientia Sinica 2017;25(4):420-424
Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.
4.Establishment and application of a Taqman MGB real-time PCR for the detection of Streptobacillus moniliformis
Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE ; Fangwei DAI ; Xiaoying SA ; Jiejie DAI
Chinese Journal of Comparative Medicine 2015;(8):62-67
Objective To establish a real-time quantitative PCR ( qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals .Method According to the S. moniliformis sequences published in NCBI , we designed specific primers and MGB probe .The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains .Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews , were detected by this established Taqman MGB qPCR method .Results We had successfully established the S.moniliformis Taqman MGB qPCR method . S.moniliformis was not detected in the samples of mice , rats, guinea pigs, hamsters and rabbits.The positive rate of S. moniliformis was 1.5% ( 1/65 ) and 61.7% ( 37/60 ) in conventional Mongolian Gerbils and tree shrews , respectively . Conclusions Our developed qPCR method can be used to effectively detect S.moniliformis in laboratory animals .Moreover , its accuracy and sensitivity are better than the national standard method .This study laid the foundations for optimizing the quality inspection system of laboratory animals .
5.Isolation and chatracterization of microsatellite markers in Tupaia belangeri chinensis
Yuan ZHANG ; Xiaofei LI ; Zhenyu LI ; Pinfen TONG ; Lingxia CHEN ; Bowen YIN ; Jiejie DAI
Chinese Journal of Comparative Medicine 2015;(6):36-41
Objective To screen out specific microsatellite markers for use in Tupaia belangeri chinensis genetic testing. Methods Firstly to screen about 700 microsatellite loci from whole genome.Secondly to choose about 100 better loci without defect factors.Lastly 46 primers were designed by 33 tree shrew’ s microsatellite loci obtained from whole genome and other references.Agarose gel electrophoresis and polyacrylamide gel electrophoresis were used for PCR products, and better loci based on electrophoresis results were chosen.Then STR scan was used to select the microsatellite loci combination for genetic testing.Results Twenty-two microsatellite loci were selected with a significant Stutter peak on STR scanning.Comparing the alternative loci and ultimately selected loci, there were two loci available in the five alternative loci of T.glis.The coincidence rate between T.glis and T.b.chinensis was 40%.There were two loci available in the five alternative loci of T.minor, and the coincidence rate between T.minor and T.b.chinensis was 40%.There were two loci available in the three alternative loci of T.belangeri, and the coincidence rate between T.belangeri and T.b. chinensis was about 70%.Conclusions The 22 microsatellite loci screened in this study are well applied for genetic testing of Tupaia belangeri chinensis, therefore, provide a scientific basis for the genetic quality monitoring of tree shrews.
6.Establishment and application of a RT-nPCR assay for detection of orthoreovirus in tree shrews
Xiaofei LI ; Anguo YIN ; Yuan ZHANG ; Jun LUO ; Xiaomei SUN ; Jiejie DAI
Chinese Journal of Comparative Medicine 2014;(6):63-68
Objective To establish a reverse transcription nested polymerase chain reaction ( RT-nPCR ) assay for detection of tree shrews orthoreovirus (TRV).Methods Three strains of TRV were respectively isolated from fresh feces of three tree shrews that came from the same field at different times .We designed and synthesized two pairs of MRV L1 gene nested primers and established the system of RT-nPCR.The TRV RNA was extracted and reversely transcribed to cDNA as a template for nested-PCR amplification.The developed RT-nPCR was optimized.The specificity and sensitivity were tested.Finally, the RT-nPCR was used to detect TRV in 25 tree shrew samples.Results Taking the genomic RNA of TRV as template, the RT-nPCR was able to amplify a specific fragment band targeting the L 1 gene, while there were no target bands in the normal cell control , ( Wa strain rotavirus , hepatitis A virus , and herpes simplex virus ) .The RNA of TRV was diluted by 1:10 to 1:109 .Each dilution sample was analyzed by the RT-nPCR.The minimum detectable concentration of RNA was 0.01 pg/μL.The results of RT-nPCR detection showed that 4 of the 15 tree shrews were TRV-positive in the survival group , and 10 of 10 tree shrews were TRV-positive in the death group . Conclusions The RT-nRCR assay established in this study is accurate , specific and sensitive .Therefore, it can be used for routine detection of TRV in quality assurance testing .
7.Histological observation of the major endocrine organs in laboratory tree shrews
Dexuan KUANG ; Xiaomei SUN ; Caixia LU ; Wenguang WANG ; Pinfeng TONG ; Anguo YIN ; Xiaofei LI ; Jiejie DAI
Chinese Journal of Comparative Medicine 2014;(6):35-39
Objective To understand the histological characteristics of the major endocrine organs of tree shrew , and provide a normal histological atlas of endocrine organs of tree shrew .Methods Ten artificially fed healthy tree shrews were killed and dissected after anesthesia .The thyroid, parathyroid, adrenal and pituitary glands were observed by gross inspection and samples were taken for routine histological examination with HE staining .Results ( 1 ) The thyroid gland was pale yellow, located on both sides of the 2-4 tracheal rings.The thyroid gland was plate-shaped, its surface was covered with a thin fibrous capsule . The thyroid parenchyma was divided into several lobules by stretched capsule membrane .Follicular and parafollicular cells were distributed in the lobules , and red colloid was present in follicular cavity.(2) Each side had one parathyroid , located on the cranial or the outer surface of the middle part of the thyroid gland, and was slightly covered by thyroid .The gland was round or oval , and its parenchyma was made up of the principal cells and eosinophil cells , and acinar structure appeared in the parenchyma .( 3 ) The adrenal glands were oval , yellow color, located in the renal hili , and linked to the kidneys .They were surrounded by a thin capsule .The parenchyma was divided into cortex and medulla .The cortex was divided into zona glomerulosa , zona fasciculata and zona reticularis from outside to inside.The zona glomerulosa was the thickest layer and the zona fasciculata was the thinnest .The medulla cells formed clumps or mesh, with central vein in the central part .(4) The pituitary gland was located in the sella turcica , with no recessus hypophysis .The pituitary gland was composed of the adenohypophysis and neurohypophysis .Its surface was covered with a connective tissue capsule .The pituitary gland was divided into distal part , middle part and pars tuberalis . neurohypophysis was made up of neural and pars infundibularis .Conclusions The histological atlas of endocrine organs in the tree shrew is established , which is close to that of the primate animals in the morphology , and provide histological evidence for the study of tree shrew endocrine organs and disorders , as well as the animal model of human diseases .
8.Isolation and identification of two different genotypes of Tupaia orthoreoviruses
Jiansheng LIU ; Yufen TAO ; Xiaofei LI ; Chao LI ; Xiaofei LI ; Xiaomei SUN ; Jiejie DAI ; Hongqi LIU
Acta Laboratorium Animalis Scientia Sinica 2015;(4):365-370
Objective To isolate and identify viruses from fecal samples of tree shrew with diarrhea.Methods Fecal sample supernatant of tree shrew with diarrhea was inoculated to three cell lines ( Vero, LLC-MK2 and KMB17 ) , and the cytopathic effects on the cells were observed.The infectious particles in the culture supernatant were further ana-lyzed by transmission electron microscopy ( TEM) , genomic RNA-PAGE, rotavirus detection kit, amplification of S1 com-plete segment and bioinformatics analysis.Results Constant cytopathic effects were induced in Vero, LLC-MK2 and KMB17 cell lines after three passages of culture.The results from TEM, RNA-PAGE and rotavirus analysis indicated that they belong to reoviruses.Analysis of the S1 segments revealed that the S1 sequence from KMB17 cell culture had the high-est homology with that of prototype isolate T1L (85%nucleotide homology and 90%amino acid homology), therefore this isolate was named as type I reovirus.The other two S1 sequences from LLC-MK2 and Vero cell culture were identical to have 85%nucleotide homology and 92%amino acid homology with the prototype isolate T3D, named as type III reovirus. Phylogenetic analysis indicated that the isolates in this study are evolutionally adapted to tree shrews.Conclusions It is the first report here that 2 genotypes of Tupaia orthoreovirus are isolated and identified from one fecal sample via three cell lines and viral S1-specific primers, which provides useful guidelines for the isolation and identification of other reoviruses from tree shrew or other hosts.
9.Progress in application of tree shrew models in research on human viral diseases
Anguo YIN ; Dexuan KUANG ; Xiaofei LI ; Yuan ZHANG ; Xiaomei SUN ; Xueshan XIA ; Jiejie DAI
Acta Laboratorium Animalis Scientia Sinica 2014;(2):86-89
Tree shrews get more and more concerns due to many of its physiological , biochemical and anatomical characteristics similar to those of human beings .Therefore, tree shrews models of human diseases such as viral diseases , neurological diseases and tumors attract more and more attention of researchers .In this article we will review the recent ad-vances in application of tree shrew models in research on human viral diseases .
10.Research progress of tree shrew models of viral hepatitis and modeling strategy
Yiquan PANG ; Yue FENG ; Xiaomei SUN ; Li LIU ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2014;(2):95-110
Viral hepatitis is a major liver disease caused by virus infection .Viral hepatitis is popular in China , mainly caused by hepatitis B and hepatitis C viruses .Experimental animal model is a necessary platform for the research on mechanism of viral infection and pathogenicity , for treatment and vaccine development .Up to date, a great progress in the development of viral hepatitis animal models has been achieved in spite of the most of findings are limited to hepatitis B and C.Here, we summarize the recent findings of viral hepatitis animal models , focusing on the tree shrew animal model and its modeling strategy .