3.Establishment of a rapid method for synthesis and detection of gRNA
Jianyong DU ; Ran DENG ; Hong GAO ; Weidong YONG
Acta Laboratorium Animalis Scientia Sinica 2015;(3):267-271,290
Objective The purpose of this study was to establish a rapid method for synthesis and detection of guild RNA (gRNA) which is an essential component in CRISPR/Cas9 knockout technology .Methods First, the Nkp46 gRNA core fragment was synthesized as amplification template .Second , the forward and reversed primers of the matched gRNA were designed using the Nkp46 gene as reference sequence .Third, the DNA fragment of Nkp46 gRNA was amplified by PCR technology using the synthesized gRNA core fragment as template .The gRNA was reversely transcribed in vitro u-sing amplified DNA fragment as template .The efficiency and specificity of gRNA and its interaction with Cas 9 were detec-ted in vitro.Results The specificity and activity of Nkp46 gRNA were high .The obtained gRNA interacted with Cas 9 en-zyme and successfully cut the target double-stranded DNA at the designed site .Conclusions The method for synthesis and detection of gRNA established in this study is faster than the original method , and the created gRNA is fully functional .
4.Evaluation of HCV-IgG,HCV-cAg and HCV-RNA in the diagnosis of Hepatitis C
Hong WU ; Yong XU ; Jinhong ZHU ; Xiaoman WU ; Xiaoyan DENG
International Journal of Laboratory Medicine 2017;38(14):1890-1891,1894
Objective To evaluate the diagnostic value of hepatitis C virus core antigen(HCV-cAg),hepatitis C virus(HCV-IgG) and hepatitis C virus(HCV-RNA) in the laboratory diagnosis of Hepatitis C.Methods HCV-cAg and HCV-IgG were detected by enzyme-linked immunosorbent assay(ELISA),HCV-RNA was detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR) in 84 suspected HCV patients and 87 healthy control subjects.Results In 84 suspected HCV patients,the HCV-IgG positive rate was 84.5%,HCV-cAg positive rate was 13.1%,HCV-RNA positive rate was 52.4%.Among 71 cases of HCV-IgG positive patients,there were 35 cases with negative HCV-RNA,the false positive rate was 49.3%.In 11 cases of HCV-cAg positive patients,there were 5 cases with negative HCV-RNA,the false positive rate was 45.5%.In 44 cases of HCV-RNA positive diagnosis of hepatitis C patients,HCV-IgG false negative rate was 18.2%,HCV-cAg false negative rate was 86.4%.The false negative rate of combined detection of HCV-cAg and HCV-IgG was 13.6%,and the true positive rate was 100.0%.Conclusion HCV-cAg and HCV-IgG have certain false negative and false positive in laboratory diagnosis of HCV,combine these two methods,or joint with HCV-RNA detection,could reduce the rate of missed diagnosis.
5.Antisense phosphorothioate oligonucleotide targeting bFGF enhances chemosensitivity of human laryngeal squamous carcinoma cells
Hong-Liang HUANG ; Hong WANG ; Jun-Jian XIANG ; Yong TANG ; Ning DENG ; Hong-Yu YANG ;
Chinese Journal of Cancer Biotherapy 2006;0(05):-
Objective:To study the enhancing effect of bFGF-targeted antisense phosphorothioate oligonucleotide (APO)on the chemosensitivity of human laryngeal squamous carcinoma cell line Hep2 to Doxorubicin,5-Fluorouracil, and Cisplatin.Methods:bFGF-specific APO was designed,constructed and transfected into Hep2 cells with jetPEI (polyethyleneimine).Expression of bFGF mRNA was evaluated by semi-quantitative RT-PCR after transfection;immuno- cytochemical method was used to examine the expression of bEGF expression before and after transfection of Hep2;the in- duction of cell apoptosis was analyzed by flow cytometry;cell proliferation was then analYzed by MTT assay after treatment with bFGF-specific APO or chemotherapeutic drugs,or a combination of both.Results:bFGF-specific APO inhibited the growth of Hep2 cells in a dose-and time-dependent manner,with the peak inhibitory rate being 25.5%.The expression of bFGF mRNA and protein decreased by 52.0% and 41.1%,respectively.The apoptosis rate of Hep2 cells was 20.5% after transfection,bFGF-specifie APO reduced the 50% inhibitory concentration of Doxorubicin,5-Fluorouracil,and Cis- platin in Hep2 cells by 75.5%,83.5% and 65.4%,respectively.Conclusion:bFGF-specific APO can enhance the chemosensitivity of Hep2 cells,which paves a new way for potential biologic chemotherapy of laryngeal squamous carcino- ma.
6.Research progress of protein tyrosine phosphatase SHP-2.
Journal of Zhejiang University. Medical sciences 2012;41(5):581-585
The Src homology-2 domain-containing phosphatase SHP-2 encoded by PTPN11 is an essential component in several signaling pathways.Different types of mutation in SHP-2 have been confirmed in several types of leukemia and solid tumors. Elucidation of the events underlying Shp2-evoked transformation may provide new insights into the novel targets for anti-cancer therapy.
Humans
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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chemistry
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metabolism
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physiology
7.Mesenchymal chondrosarcoma of thoracic spine: report of a case.
Xiang-lei HE ; Deng PAN ; Yu ZHOU ; Yong-li GAN ; Hong-bin ZHANG
Chinese Journal of Pathology 2006;35(12):766-767
12E7 Antigen
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Antigens, CD
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metabolism
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Cell Adhesion Molecules
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metabolism
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Child, Preschool
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Chondrosarcoma, Mesenchymal
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metabolism
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pathology
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surgery
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Diagnosis, Differential
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Humans
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Immunohistochemistry
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Male
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Neuroblastoma
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metabolism
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pathology
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Sarcoma, Ewing
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metabolism
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pathology
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Spinal Cord Neoplasms
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metabolism
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pathology
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surgery
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Spinal Neoplasms
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metabolism
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pathology
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Thoracic Vertebrae
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Vimentin
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metabolism
8.Adult extrarenal Wilms' tumor occurring in ovary: report of a case.
Li LIANG ; Xin-hua ZHOU ; Yong-jian DENG ; Hong-hai ZHANG ; Yan-qing DING
Chinese Journal of Pathology 2008;37(4):284-285
Female
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Humans
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Kidney Neoplasms
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complications
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pathology
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Ovarian Neoplasms
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pathology
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Wilms Tumor
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complications
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pathology
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Young Adult
9.Distribution Characteristics and Drug Resistance Analysis of Pathogens from 2005 to 2007
Guoping YANG ; Chao DENG ; Hong YUAN ; Zhijun HUANG ; Min SONG ; Yong QI
Chinese Journal of Nosocomiology 2009;0(23):-
OBJECTIVE To investigate the uses of antibacterials,the characteristics of bacterial distribution and the drug resistance and provide a foundation for reasonable application of antibacterials.METHODS Germs isolation and cultivation were carried out according to National Clinical Testing Operation Procedures(2nd edition).Germs definition and resistance test were carried out by using VITEK 2 Compact microbiological assay system.Statistical analysis was processed by using WHONET 5.4.RESULTS Totally 7815 strains of bacteria were isolated from clinical samples,most of which were Gram-negatives(G-)(58.0%,56.3% and 59.5%,respectively in the 3 years).The tendency of Gram-positives(G+) didnot charee obviously(about 20%).Theresistance were increasing.The resistance rate of Staphylococcus aureus was more than 70.0 % to the most antibacterials.Up to now,there was no vancomycin-resistant isolate.CONCLUSIONS We should use antibacterials reasonably and efficiently to delay and control drug resistance,according to the susceptibility test.
10.Effect of different dispensing methods on content of omeprazole sodium for injection
Guochun LI ; Xinwu HUANG ; Gulin DENG ; Hong XU ; Longmei LIU ; Yong TANG
Chinese Journal of Biochemical Pharmaceutics 2015;35(10):155-157
Objective To select omeprazole content changes smaller with dispensing method and to seek for rationality of off-label uses.Methods To measure content change of omeprazole sodium for injection mixed by different subscriptions at different time through HPLC, and compared effect of different dispensing methods on content of omeprazole sodium for injection.Results 10 mL 0.9%sodium chloride injection was chosed as dissolvent,the change of omeprazole sodium for injection content would be minor, and stability of drug solution was superior.Conclusion Dispensing methods of drug impact on its'security and validity, which is part of discuss category about medicine rational use as well.Off-label uses could not vest in unreasonable use, which should contingent on specific document,data and actual environment of medical treatment.