2.Infectious Cloning Approach for Herpesvirus Based on Bacterial Artificial Chromosomes
Jian-Hong LU ; Yun-Lian TANG ; Gui-Yuan LI ;
China Biotechnology 2006;0(06):-
The genetic analysis of herpesviruses has been a constant challenge, due to the large, complex genomes of herpesviruses and mutagenesis of viral genes by conventional recombination methods in cell culture. Recently, a completely new approach for full-length infectious clones of herpesviruses based on bacterial artificial chromosomes (BACs) has been developed. This technique allows the maintenance, propagation and genetic modification of the viral genome as a BAC plasmid in E.coli, thus making the procedures fast, safe and effective in prokaryotic cells. This technique also makes it possible for the reconstitution of viral progeny or mutants by transfection of the BAC plasmid into eukaryotic cells, thereby facilitating the analysis of viral gene functions in the context of genome. In this presentation, Epstein-Barr virus was used as an example to describe the principle, establishment of the technique and mutation introduction into the BAC plasmid, and to discuss the perspective in the use of BAC-cloned herpesviruses.
3.Effects of MMP-7 mRNA expression in esophageal cancer
Jian-Hong LIAN ; Chun-Li WANG ; Wei-Lan LIU ; Rui-Hong YANG ; Su-Tang GUO ;
Cancer Research and Clinic 2006;0(11):-
Objective To investigate the relationship between the expression of MMP-7 in peripher- al blood and esophageal cancer.Methods By using Real time RT-PCR, blood samples from 82 cases with postoperative esophageal cancer were detected.Results 37.8 %(32/82)patients were positive for MMP-7 mRNA expression.MMP-7 mRNA positivity significantly correlated with lymph node metastasis,stage,and hematogeneous metastasis.Recurrent disease was found in 28 of 82 cases. Of 28 patients experiencing re- lapse,15 patients were serially follow-up in every three months.The number positive of MMP-7 mRNA be- fore detection by imaging,at the same time of detection by imaging,and after detection by imaging was 5 cas- es,8 cases and 2 cases respectively.Conclusion Examination of MMP-7 mRNA in peripheral blood during follow-up is useful for early detection of occult recurrence.
4.The entry of Epstein-Barr virus into B lymphocytes and epithelial cells during infection.
Lie-Lian ZUO ; Mei-Juan ZHU ; Shu-Juan DU ; Jian-Hong LU ; Gui-Yuan LI
Chinese Journal of Virology 2014;30(4):476-482
Epstein-Barr virus (EBV) is a human herpesvirus associated with important human diseases, including infectious mononucleosis syndrome, malignant lymphoma, and nasopharyngeal carcinoma. The mechanism of EBV entry into host cells remains a subject of intensive research. After decades of study, researchers have identified several key proteins and different patterns of EBV intrusion into host cells. The viral surface glycoproteins, gp350/220, gp42, gB, gH, and gL, are involved in interactions with the CR2 receptor on the surface of B lymphocytes during viral entry. However, the majority of epithelial cells lack CR2 receptor expression, which makes viral invasion much more complex than in B lymphocytes. Three different models have been proposed to explain how EBV enters epithelial cells: (1) "transfer of infection", mediated by B lymphocytes or Langerhans cells; (2) EBV utilizes its own proteins during the process of fusion with the cell membrane; and (3) progeny virions arising from EBV-infected epithelial cells cross lateral membranes into adjacent epithelial cells. This review will discuss the relevant mechanism of viral entry into B lymphocytes and epithelial cells during EBV infection.
Animals
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B-Lymphocytes
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virology
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Epithelial Cells
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virology
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Epstein-Barr Virus Infections
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virology
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Herpesvirus 4, Human
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genetics
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physiology
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Humans
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Viral Proteins
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genetics
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metabolism
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Virus Internalization
5.Application of Color Doppler Ultrasonography by Bed Side in the Early Diagnosis of Hypoxic-Ischemic Encephalopathy in Full Term Neonates
yi-jin, SU ; lian-fang, DU ; jin, XIA ; min, FANG ; xian-ming, XU ; jian-guo, HONG
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.
6.Direct spectrophotometric method to determing serum copper with a new water soluble reagent
Tie-Li ZHOU ; Chang-Rong ZHAO ; Hong-Wu XIAO ; Guo-Jun LIAN ; Jian-Ming CAO ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To establish a simple and sensitive method for the determination of serum copper by spectrophotometry.Methods Nitro-PAPS was used as a coloring agent for serum copper in the presence of surfactants Tween-80 and Triton X-100 and the formed complex was measured by spectrophotometry.Results The maximum absorption wavelength of the complex was 570 nm and the molar absorption coefficient was 7.95?10~4 L/(mol?cm).The lineafity of the method was up to 63.0 ?mol/L and the recoveries ranged from 98.6% to 103.1%.The within-run and between-run CVs were 2.1%-3.3% and 2.7%-3.8%.The method(Y)was compared with an AAS method(X)and a correlation of Y=1.01X -0.27(r=0.998 2)was obtained.A reference interval(x~-?2s)determined with this method on 68 individuals was 9.7-24.1 ?moL/L.Conclusions A simple and sensitive method for serum copper has been established.It may used for the analysis of serum copper in clinical laboratories.
7.Molecular Cloning and Sequence Analysis of CAPN1 Gene in Yak
Chun-Hong FEI ; Jian-Ping WU ; Lian YANG ; Li-Ping ZHANG ; Jia WANG ;
China Biotechnology 2006;0(12):-
96%).There are 14 nucleotide mutations between yak and cattle and 3 of that were predicted to alter the protein sequence,both of which occurred in the domain III.Construction of molecular phylogenetic tree shows:The cluster results coincided with the traditional taxonomy.
8.Identificition of Endophytic Bacteria EBS05 from Cinamonum camphra and Property of Its Antimicrobial Compound
Cai-Yi WEN ; Zhi-Gang YIN ; Jian-Guang CHEN ; Hong-Lian LI ;
Microbiology 2008;0(07):-
An endophytic bacterium strain EBS05 from Cinamonum camphra was identified as Bacillus subtilis by morphological taxonomy and sequence analysis of 16S~23S rRNA intergenic spacer regions. Properties of antimicrobial compound produced by EBS05 were assayed. The active compound had the maximum absorbance peak at ?213.5 nm. The antimicrobial activity was stable in solution with pH value from 5 to 8, and decreased significantly in solution with pH value less than 4.0 or more than 9.0. The antimicrobial compound had thermodynamics stability. Its activity changed a little after treated at 60?C~80?C for two hours, and compared with 65% original activity after treated at 1?105 Pa for 30 minutes. The active substance had high resistance to ultraviolet radiation and protease K. Antimicrobial compound was soluble in alcohol solu- tion, which was easily dissolved in methanol and ethanol, but not dissolved in ethyl acetate, acetonitrile and petroleum et al.
9.Relationships between Serum Lipid,Apolipoproteins and Angiocardiopathy in Obese Children
shi-lian, LI ; mei-he, ZHANG ; wen-qi, SONG ; jian-yun, JIN ; qiu-hong, ZHAO ; huan-ling, YU
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To analyze obese children serum lipid level in order to understand the relationship between serum lipid and cardiovascular disease in obese children.Methods One hundred and fifty-three children(109 male and 44 female)aged 4-16 years old with obesity who attended the outpatient clinic of Beijing Children′s Hospital were collected.Percentage body fat (%BF),body fat (BF),fat-free mass (FFM) was estimated by using bioelectrical impedance analysis (BIA) and calculate.Waist and hip circumference,waist-to-hip ratio (WHR) was estimated by soft tape measure and calculate.Skinfold thickness of scapular bone below (S) and triceps muscle (T),S/T rate was estimated by skin fold meter and calculate.Serum total cholesterol (TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),apolipoprotein(Apo) AI and Apo B levels were also measured.SPSS 11.0 software was used to analyzed the data.Results The cardiovascular disease related was the prevalence of high TC levels(3.3%)or high LDL-C level(6.0%) and high TG level(24.7%) was rather low.HDL-C level was reduced in 31.3% of obese children.In children over 10 years old,%BW and %BF showed a weak correlation with HDL-C(r=-0.202,-0.211).Conclusions In obese children,serum lipid as well as Apo level should be exa-mined in order to evaluate angiocardiopathy.
10.Influence of mesenchymal stem cells implantation on ventricular remodeling and heart function after acute myocardial infarcion
ri-tai, HUANG ; hong-sheng, ZHU ; song, XUE ; feng, LIAN ; gang, HUANG ; jian-jun, LIU ; ru-qi, TAO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To study the influence of mesenchymal stem cells(MSCs) implantation on ventricular remodeling and heart function after myocardial infarcion. Methods Bone marrow was aspirated from Gui-zhou Xiang swines.After being isolated,cultured and co-cultured with 5-azacytidine,either autologous MSCs(experiment group) or a comparable volume of physiologic saline(control group) were injected into the infarcted myocardium.Three and six weeks later,echocardiographic measurement was performed to assess the myocardial structure and heart function,and single photon emission computed tomography(SPECT) was employed for myocardial imaging.Implanted stem cells were detected by the anti-Brdv antibody DAB with HE staining. Results Left ventricular ejection fraction(EF),fractional shortening and wall thickening were higher in experiment group than control group.The thickness of the ventricular wall and septum was found increased while the left ventricular chamber size was smaller in experiment group.It was indicated by SPECT that three and six weeks after implantation,there was obvious image defect in control group while in experiment group there were some imaging areas in the infarcted area.Brdv-labelled cells were observed in the central part of and around the infarcted area.Conclusion Implantation of MSCs into the infarcted myocardium is believed to attenuate the remodeling process,inhibit the extent of wall thinning and dilatation of the ventricular chamber.MSCs implantation may also improve the contractile ability of the myocardium and heart function.