1.Suckling mouse model establishment of myocarditis induced by foot-and-mouth disease vi rus
Keshan ZHANG ; Jijun HE ; Jiajun WU ; Yongjie LIU ; Weiwei CHENG ; Youjun SHANG ; Xiangtao LIU ; Xuepeng CAI
Chinese Journal of Zoonoses 2014;(10):987-989,996
Interactions between FMDV and cardiac cells are multifaceted and complex ,these interactions leads to pro-teins alterations in cardiac cells inevitably .To understand the pathogenesis of myocarditis after FMDV infection in mice ,the suckling mouse model for myocarditis induced by foot-and-mouth disease virus (FMDV) was established in this study .Suckling mice within 3 days old was selected to infect by FMDV .Myocarditis caused by FMDV in suckling mice was confirmed with clinical symptom monitor .The observation of Hematoxylin and eosin stain (H&E stain) and transmission electron microscopy (TEM) were performed after samples processing .According to conventional polymerase chain reaction (PCR) methods ,prim-ers of VP1 gene was designed ,synthesised and specific FMDV VP1 gene was amplified from the heart muscle of suckling mice . The results indicated that suckling mice appeared low spirit condition ,dyspnea ,and dull reaction within 36 hours after chal-lenge with FMDV .Infiltration of inflammatory cells and dissolution of myocardial fibers were observed with H&E stain and TEM .Special target gene of FMDV was amplified from the heart of infected group .Obvious inflammation in the heart of suck-ling mice caused by FMDV was observed .It's suggested that suckling mouse model for myocarditis induced by FMDV was es-tablished successfully ,which would lay the foundation for researches of myocarditis mechanism in young cloven-hoofed ani-mals .
2.Expression of CD3+ CD8+ human leukocyte antigen-A2+ T lymphocytes with specificity to the different hepatitis B virus peptides in patients with hepatitis B associated hepatocellular carcinoma
Jilin CHENG ; Liwei WANG ; Chenli QIU ; Yingchun AI ; Jihua LU ; Keshan YIN ; Shaoping HUANG ; Rong TANG ; Lie XU ; Yi ZHANG
Chinese Journal of Infectious Diseases 2012;30(5):264-267
ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44%±0.04%,which was higher than those to other four HBV antigen peptides (0.68%±0.08% of FLLTRILTI,1.06%±0.09% of FLPSDFFPSV,0.56% ±0.04% of FLLSLGIHL,and 0.46% ±0.08% of WLSLLVPFV) (t=0.001,P<0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.
3.CD4 and CD8 cells in gastric mucosa from AIDS patients with gastritis infected by Helicobacter pylori
Duoduo LI ; Liwei WANG ; Ye ZHENG ; Dong ZENG ; Shaoping HUANG ; Zhenyu FAN ; Yahong XU ; Jihua LU ; Keshan YIN ; Rong TANG ; Jie CAO ; Renfang ZHANG ; Yanling FENG ; Jilin CHENG
Chinese Journal of Microbiology and Immunology 2012;(11):949-952
Objective To investigate the relationship among Helicobacter pylori(H.pylori),CD4 positive cells and CD8 positive cells in gastric mucosa of the AIDS patients with gastritis.Methods Fiftyeight AIDS patients with upper abdominal pain were diagnosed with chronic gastritis through gastroscopy.The gastric biopsies from them were used for H.pylori detection with rapid urease test and Giemsa staining,pathology examination with HE staining,and immunohistochemistry analysis for CD4,CD8 positive cells in Gastric mucosa.And the application of flow cytometry was for the detection of peripheral blood CD4 and CD8 lymphocytes from the patients.Results H.pylori was positive in 26 cases,and negative was in 32 cases.CD8 cell expression in gastric mucosa of the AIDS patients with H.pylori positive was significantly higher than H.pylori negative patients(P<0.05).There is no difference CD4 cell expression in gastric mucosa between the AIDS patients with H.pylori positive and H.pylori negative patients.Moreover,CD8 positive lymphocytes in gastric mucosa of those patients with H.pyloriinfection were significantly stronger than the CD4 positive lymphocytes.However,the peripheral blood CD4 lymphocytes from the patients with H.pylori infection were more than those from H.pylorinegative patients significantly(P<0.05).Conclusion The expression level of CD8 cells in gastric mucosal tissues of AIDS patients with H.pylori infection were higher than those without H.pylori infection.The CD4 lymphocytes from the peripheral blood of the patients with H.pylori infection were more than those without H.pylori negative patients.
4. Mitochondrial quality control system as a therapeutic target for treating heart failure
Junfan LI ; Shuqiu SUN ; Yingxue WANG ; Kunyu ZHANG
Chinese Journal of Endemiology 2019;38(12):1028-1032
The mitochondrial quality control system maintains mitochondrial homeostasis mainly through protein degradation, vesicle transport, and mitophagy. Mitochondrial biosynthesis, dynamics, and calcium ion play key regulative roles in mitochondrial quality control. Under normal conditions, the mitochondrial quality control system can work well. In recent years, studies have found that mitochondrial dysfunction is closely associated with the occurrence of heart failure. In order to understand mitochondrial function, this paper reviews mitochondrial quality control methods, regulatory factors and their potential therapeutic applications in heart failure.
5.Preparation of monoclonal antibodies against VP2 protein of Asia I type foot-and-mouth disease virus and establishment of a competitive ELISA for the detection of antibodies.
Min XIANG ; Keshan ZHANG ; Shun LU ; Lijun CAI ; Yong LUO ; Jianmin ZHANG ; Hua HE ; Qingang WANG ; Bin WU
Chinese Journal of Biotechnology 2008;24(9):1664-1669
Monoclonal antibodies against FMDV vp2 protein were prepared and a competitive ELISA based on the monoclonal antibodies and vp2 protein was established. Balb/c mice were immunized with Escherichia coli expressed fusion protein. The splenocytes from immunized mice were fused with myeloma cells SP2/0. The hybridism cells were screened by indirect ELISA and limited dilution method. Two hybndoma cell Iines secreting mAbs against Asia I type foot-and-mouth disease were obtained. The titer and relative affinity of mAbs were determined by ELISA. Specificity of mAbs was analyzed by Western blotting. The ELISA titers of the ascites induced by the two hybridism cells were above 100 x 2(9).A competitive ELISA for the use of FMDV antibody detection was established using E. coli expressed fusion protein as coating antigen and HRP-labled mAb as detecting antibody. Clinical tests showed the method had 89.0 percent agreement with UBI Kit to detection of FMDV antibodies and 86.5 percent agreement with LPB- ELISA kit (Ceditest kit) for detection of antibodies against Foot-and-Mouth Disease Virus respectively.
Animals
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Antibodies, Monoclonal
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biosynthesis
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immunology
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Antibodies, Viral
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blood
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Capsid Proteins
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immunology
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Enzyme-Linked Immunosorbent Assay
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methods
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Female
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Foot-and-Mouth Disease
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immunology
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virology
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Foot-and-Mouth Disease Virus
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immunology
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Mice
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Mice, Inbred BALB C
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Swine
6. Efficacy and safety of low dose sublingual nifedipine dripping pills (5 mg) in the acute treatment of moderate and severe hypertension: a randomized, double-blind, positive-drug parallel-controlled, multi-center clinical study
Jihai LIU ; Yaling HAN ; Shuyang ZHANG ; Yan WEI ; Zhanquan LI ; Yukai WANG ; Yao QING ; Ying HUANG ; Xiaoping CHEN ; Ximing CHEN ; Hong WANG ; Yingjie LI ; Yunqiu MO ; Danming WU ; Keshan LIANG
Chinese Journal of Cardiology 2019;47(5):374-380
Objective:
To evaluate the efficacy and safety of low dose sublingual nifedipine dripping pills (5 mg) in treating moderate and severe hypertension in comparison with normal dose (10 mg) of sublingual nifedipine dripping pills.
Methods:
This study was designed as a randomized, double-blind, positive drug parallel controlled, multi-center, non-inferiority clinical trial. Patients with moderate and severe hypertension were enrolled by 14 clinical trial centers, randomly divided into the trial group (sublingual 5 mg nifedipine dripping pills) and the control group (sublingual 10 mg nifedipine dripping pills). The changes in blood pressure were monitored continuously within 2 hours after the initial administration, repeated the dose in 20 minutes interval after the initial administration for up to additional 3 doses (maximum 4 doses) if the antihypertensive efficacy was not satisfactory. The efficacy of antihypertensive therapy between the two groups was evaluated by repeated administration rates and blood pressure changes at 60 minutes post the initial administration, and the safety of treatment was evaluated by recording adverse event rate of the two groups.
Results:
The anti-hypertensive effective rates at 60 minutes after sublingual administration were 83.5% (202/242) and 86.7% (208/240) respectively between the trial group and control group (χ2=1.307,