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.Establishment of a EV71 virus infection model of tree shrew primary renal cells
Ming YANG ; Xiaoxing HU ; Wenguang WANG ; Li ZHANG ; Shuwei DONG ; Yue FENG ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2017;25(2):117-122
Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.
6.Establishment and application of CODEHOP PCR assay for detection of Pasteurella spp.in laboratory animals
Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE ; Xiaomei SUN ; Jiejie DAI
Chinese Journal of Comparative Medicine 2017;27(1):85-90
Objective We established a rapid detection method of Pasteurella spp.and provided a reference for microbiological quality control of laboratory animal .Methods According to the β subunit of bacterial RNA polymerase ( rpoB) protein multiple alignments of 13 different Pasteurella spp.published in NCBI .The degenerate primers were designed by CODEHOP designer online .CODEHOP PCR method was applied to detecting Pasteurella spp.after the specificity and sensitivity of the method had been evaluated by 21 reference strains .Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5.The primers are able to distinguish between Pasteurella spp.and the other pathognic organisms of laboratory animal respiratory tracts .Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella.The Pasteurella positive rate was 19.1% in 609 animal ' s respiratory samples .The accuracy of positive results was 100%through verifying by sequenced and blast .Conclusions The established method has good specificity and sensitivity .It can be used to detect Pasteurella spp.in animal samples .
7.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.
8.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 .
9.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 .
10.Isolation,culture, adipocgenic and osteogenic induction of Tupaia bone marrow mesenchymal stem cells
Caixia LU ; Xiaofei LI ; Wenguang WANG ; Xiaomei SUN ; Pinfen TONG ; Dexuan KUANG ; Jiejie DAI
Chinese Journal of Comparative Medicine 2014;(3):10-13
Objective To study the isolation,culture, adipogenic and osteogenic induction Tupaia bone marrow mesenchymal stem cells(BM-MSCs).Method The BM-MSCs from tupaia were isolated and expended by combination of gradient centrifugation and adherence culture , then subcultured and observed for morphology under inverted phase contrast microscope.BM-MSCs were induced to adipocytes .and osteoblasts in vitro Result Cells were spindle or triangle-shaped, and clone proliferation .Cells were successfully induced into adipocytes .and osteoblasts Conclusions The method of isolation BM-MSCs from tupaia by combination of gradient centrifugation and adherence culture is simple and feasible , BM-MSCs have differentiation potential into adipocytes and osteoblasts .