3.Clinical observation of biopsy by double paths for higher positive percentage.
Hong-Quan WEI ; Gang YU ; Xiao-Tian LI ; Zhong REN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005;40(1):67-68
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Biopsy
;
methods
;
Child
;
Child, Preschool
;
Endoscopy
;
Female
;
Humans
;
Larynx
;
pathology
;
Male
;
Middle Aged
;
Nasopharynx
;
pathology
;
Young Adult
5.Serodiagnosis of trichinellosis by ELISA using recombinant nudix hydrolase of Trichinella spiralis
Shao Rong Long, Xiang Yu Tian, Zhong Quan Wang
Tropical Biomedicine 2015;32(4):669-675
Trichinella spiralis nudix hydrolase (TsNd) gene encoding a 46 kDa protein was
expressed in Escherichia coli and the potential of recombinant TsNd protein (rTsNd) as an
antigen for the serodiagnosis of trichinellosis was investigated by ELISA and compared with
those of ELISA with T. spiralis muscle larval excretory–secretory (ES) antigens. The sensitivity
of both ELISA was 100% (30/30), for the detection of anti-Trichinella IgG antibodies in sera of
the experimentally infected mice, and the specificity of rTsNd-ELISA and ES-ELISA was 100%
(54/54) and 98% (53/54), respectively (P>0.05). Serum anti-Trichinella antibodies were firstly
detected by rTsNd-ELISA at 14 days post infection (dpi), then continued to increase with a
detection rate of 100% at 36 dpi. The anti-Trichinella antibody levels at different times after
infection were statistically different (P<0.05). The results showed that the rTsNd might be a
potential candidate antigen for specific serodiagnosis of trichinellosis. But, it needs to be
further evaluated with sera of the patients with trichinellosis and other helminthiasis.
6.Overexpression of Alcohol DehydrogenaseⅠ in Saccharomyces cerevisiae
Li-Na QIN ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.
7.Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium
Zhi-Ping LI ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.
8.Secretion and expression of vascular endothelial growth factor and interleukin-8 by SH-SY5Y human neuroblastoma cells.
Zhigang FAN ; Yu LIN ; Qiping HUANG ; Meirong LUO ; Qinghua TIAN ; Donghuo ZHONG ; Quanyi FENG ; Zezhi WU
Chinese Journal of Biotechnology 2013;29(11):1629-1643
To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.
Biomarkers
;
Cell Line
;
Cell Proliferation
;
Cellular Microenvironment
;
Humans
;
Interleukin-8
;
genetics
;
secretion
;
Neuroblastoma
;
secretion
;
Polyesters
;
chemistry
;
RNA, Messenger
;
genetics
;
Vascular Endothelial Growth Factor A
;
genetics
;
secretion
9.Detection of soluble interleukin-2 receptor and T lymphocyte subsets in silicosis.
Yu-hua ZHU ; Yue-qiu TIAN ; Guo-ying ZHU ; Hong-zhen ZHANG ; Zhong-xing CAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(1):66-66
Aged
;
Humans
;
Male
;
Middle Aged
;
Receptors, Interleukin-2
;
blood
;
Silicosis
;
blood
;
classification
;
T-Lymphocyte Subsets
;
metabolism
10.Expression changes of cardiac sinus node Cx45,Cx31.9 on sinus bradycardia model caused by dexmedetomidine in rabbit
Yu ZHU ; Lei TIAN ; Yanruo HUANG ; Jing SHI ; Yi ZHONG ; Yongqiang YIN
The Journal of Clinical Anesthesiology 2017;33(8):801-804
Objective To investigate the changes of cardiac sinus node connexin 45 (Cx45), connexin 31.9(Cx31.9) of gap junction in rabbits'' sinus bradycardia model caused by dexmedetomidine, and to discuss whether the negative frequency and negative conduction caused by Dexmedetomidine are related to the expression changes of connexin.Methods Forty-eight healthy adult New Zealand rabbits were divided into three groups (n=16).Sinus bradycardia rabbits were prepared by intravenous injecting Dexmedetomidine through ear vein.After rabbits were anesthetized by sodium pentobarbital, basic procedures were quickly completed in order to monitor MAP and ECG.Rabbits in group C were injected with normal saline.Rabbits in group D1 were injected a loading dose of dexmedetomidine 10 μg/kg for 10 min, and then continuously pumped 5 μg·kg-1·h-1 for 50 min.Rabbits in group D2 were pumped a loading dose of dexmedetomidine 60 μg/kg for 10 min, and then continuously pumped 30 μg·kg-1·h-1 for 50 min.After observation hearts were quickly removed and sinus node tissue was dissected.The average optical density of sinus node Cx45, Cx31.9 were detected by immunohistochemistry, and the genes expression were detected by real-time quantitative.Results Cx45 gene expression of group D2 showed remarkable increase than groups C and D1(P<0.05), there were no significant differences between groups D1 and C, Cx45 average optical density change were consistent with the gene expression.Cx31.9 gene expression of group D1 showed a more remarkable increase than group C(P<0.05), there were no significant differences between groups D2 and C,D1 and D2, Cx31.9 average optical density changes were consistent with the gene expression.Conclusion Dexmedetomidine increases the expression of low electrical conductivity Cx45, Cx31.9 of gap junction on rabbits'' sinus node, which is one of the possibly reasons that slow down cardiac conduction velocity in sinus node.