1.Comparison of Three ELISA Kits for the Differentiation of Foot-and-mouth Disease Virus-infected from Vaccinated Animals
Yi-mei, CAO ; Zeng-jun, LU ; Zai-xin, LIU ; Qing-ge, XIE
Virologica Sinica 2007;22(1):74-79
A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals.Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits,Ceditest(R)FMDV-NS (Ceditest(R) kit),UBI(R) FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI(R) kit) and a FMDV 3ABC-I-ELISA kitdeveloped at the Lanzhou Veterinary Research Institute.The test parameters (sensitivity and specificity) of the three kits were determined,and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits.The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest(R) kits was 98.05%,and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI(R) kits was 94.4%; the sensitivity of both Ceditest(R) and FMDV 3ABC-I-ELISA kit was 100%.However,the sensitivity of the UBI(R) kit was only 81.8%.With sera from naive or vaccinated non-infected animals,the specificity of all tests exceeded 90%.
2.Combined Therapy with Mixed Heat Shock Protein/Peptide Vaccine and Interleukin-12 Results in Enhanced Antitumor Effects
Ge SONG ; Mei YUAN ; Xuemei CUI ; Xiang SUI ; Bin ZHAO ; Shibi LU
Chinese Journal of Rehabilitation Theory and Practice 2007;13(3):252-254
Objective To investigate the antitumor immunity induced by tumor derived mixed heat shock protein/peptide(mHSPs),interleukin-12(IL-12)and Cyclophosphamide(Cy).MethodsPurified mixed HSP was prepared from tumor by S180 protein extraction and purification,SDS-PAGE,Western blot and animal experiment were applied for mixed HSPs analysis.ResultsThe proliferation of cytotoxic T lymphocyte(CTL)cocultured in the mHSPs+Cy+IL-12 group was significantly remarkable and the content of CD8+ CTLs was significantly more in comparison with the other groups(P<0.01).To the tumor bearing mice,mHSPs+Cy+IL-12 group showed partial therapeutic efficacy,the averaged survival period was over 60 d,and 90% of the mice in this group got long period tumor free survival(>90d),obvious difference(P<0.05)from the other groups.ConclusionTumor derived mixed HSPs can induce powerful antitumor immune efficacy and show favorable therapeutic efficacy.
3.Germ cell apoptosis induced by Ureaplasma urealyticum infection
XU CHEN ; LU MEI-GE ; FENG JING-SHENG ; Guo QIANG-SU ; WANG YI-FEI
Asian Journal of Andrology 2001;3(3):199-204
Aim: To study the effect of Ureaplasma urealyticum (UU) infection on germ cell apoptosis of male rats. Methods: Male rats were infected artificially with UU serotype 8 (T960) . Morphological changes of germ cells in the seminiferous tubules and the lumen of the epididymides were observed under the light microscope. Fluorescence-conjugated polyclonal antibodies to Fas and Fas ligand (FasL) were used to localize Fas and FasL. TUNEL staining of germ cells and Sertoli cells was performed by the AKPase method. TUNEL-positive rate ( % positive cells) and TUNEL-positive area (area occupied by stained cells) were analysed by KS400 Image Analysis System. The DNA laddering analysis was performed by agarose gels electrophoresis. Results: In those rats infected with UU: (1) Exfoliated germ cells were dramatically increased. Many multinucleated giant cells were found in the seminiferous tubules and the lumen of the epididymides. (2) The number of TUNEL-positive cells and the TUNEL-positive area were significantly increased.(3) The expression of Fas and FasL in germ cells and Sertoli cells was up-regulated. (4) Discrete bands of fragmented DNA were found in the testicular cells. Conclusion: In male rats, germ cell apoptosis was increased in UU infection.
4.Evaluation of methods for detection of NPM1 gene mutations in acute myeloid leukemia.
Zhi-Peng LI ; Xuan ZHANG ; Xiao-Ming ZHAO ; Qing-Ge LI ; Ya-Mei CHEN ; Quan-Yi LU
Journal of Experimental Hematology 2011;19(4):999-1004
The purpose of this study was to establish real-time based methods for detection of NPM1 gene mutation in acute myeloid leukemia (AML). Primers/probes were designed according to the clustered region of NPM1 mutations on exon 12. Two real-time PCR assays, including high resolution melting curve (HRM) and allele-specific PCR (AS-PCR), were developed and clinically evaluated with 89 AML samples, which were parallelly detected by capillary electrophoresis (CE) and sequencing. The results showed that a total of 17 mutation-positive samples were detected, including type A (15 cases), type B (1 case) and type Nm (1 case). HRM assay could detect all mutant types, and the analytical sensitivity was around 5%. In contrast, AS-PCR assay detected only 95% mutant types, but its sensitivity was as high as 0.01%. It is concluded that considering the characteristics of each method as well as the clinical evaluation results, HRM may be used for screening of NPM1 mutations at diagnosis, while the AS-PCR can be used for the MRD quantification during follow-up.
Alleles
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DNA Mutational Analysis
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Electrophoresis, Capillary
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methods
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Genome
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Humans
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Leukemia, Myeloid, Acute
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diagnosis
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genetics
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Mutation
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Neoplasm, Residual
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diagnosis
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Nuclear Proteins
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genetics
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Plasmids
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Polymerase Chain Reaction
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methods
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Sensitivity and Specificity
5.Establishment and application of the approach to detecting two biovars of Ureaplasma urealyticum in human semen.
Mei-ge LU ; Jian-li SHI ; Chen XU
National Journal of Andrology 2005;11(3):175-184
OBJECTIVETo establish the approach to detecting two biovars of Ureaplasma urealyticum (Uu) in human semen and to investigate the relationship between the two biovars of Uu infection and the quality of human semen.
METHODSBased on the 16S-23S rRNA intergenic spacer region, three pairs of primers were designed, the species specific primer and two biovars primers (Parvo primer and T960 primer). The two biovars of Uu were detected in the semen from 949 men by semen culture and PCR assay. Meanwhile, semen routine analyses were performed.
RESULTSIn the 949 subjects, 199 were Uu positive both in Uu liquid culture and PCR assay (199/949, 21.1%), of which 136 (136/199, 68.3%) were Parvo biovar, 54 (54/199, 27.1%) T960 biovar, and 9 (9/199, 4.5%) both Parvo and T960 biovars. Compared with the Parvo and the negative groups, human sperm viability was significantly decreased (P < 0.05 ) in the Uu T960 infection group. The difference of sperm motility and density had no statistic significance.
CONCLUSIONA significant correlation has been found between Uu T960 biovar infection and human sperm viability
Adult ; Humans ; Male ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; Semen ; microbiology ; Sperm Motility ; Ureaplasma urealyticum ; classification ; genetics
6.The development and application of lie detection in forensic science.
Lu WANG ; Ling YANG ; Yan GE ; Ji-Feng CAI ; Yun-Feng CHANG ; Ling-Mei LAN
Journal of Forensic Medicine 2008;24(5):365-368
Lie detection technology has been applied increasingly to investigate and solve criminal cases. This article explores the evolvement of lie detection technology in the ancient times and the application of the psychological and physiological parameters which have become more accurate with the introduction of modern polygraph. The cognitive exploration and the application of Event Related Potentials (ERPs), functional Magnetic Resonance Imaging (fMRI), and Event-Related functional Magnetic Resonance Imaging (E-R fMRI) have made detection technology focus on the brain activities, which produce more objective results by tracing the original state of lying. In summary, this article describes different types of lie detections, simple and complex, their working principles, the latest development, and the prospect of their application in forensic science.
Evoked Potentials
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Forensic Medicine
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Humans
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Lie Detection
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Magnetic Resonance Imaging/methods*
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Psychophysiology/instrumentation*
7.Designation, solid-phase synthesis and antimicrobial activity of Mytilin derived peptides based on Mytilin-1 from Mytilus coruscus.
Mei LIU ; Mei WU ; Shiquan ZHOU ; Peng GAO ; Tao LU ; Rixin WANG ; Ge SHI ; Zhi LIAO
Chinese Journal of Biotechnology 2010;26(4):550-556
As a key role in mussel defense system, Mytilin is an important antibacterial peptide isolated from the mussel serum. The structural and functional researches on Mytilin showed that the fragment connecting two beta-sheets in a stable beta-hairpin structure was probably required for antimicrobial activity. To elucidate the structural features and the antimicrobial activity of this fragment, we re-designed and synthesized two peptides corresponding to the main mimic structures of Mytilin-1 from Mytilus coruscus, we named these two peptides Mytilin Derived Peptide-1 and Mytilin Derived Peptide-2, respectively. Using a liquid growth inhibition assay, we evaluated their activity towards Gram-positive, Gram-negative bacteria and fungus. The results showed that both peptides can inhibit the growth of Gram-positive, Gram-negative bacteria and fungus. Besides, these two peptides showed high stability in heat water and human serum. These works laid the foundation for further research on the molecular mechanism of Mytilin and for further exploitation of antibacterial peptides with lower molecular mass and more stable structure.
Amino Acid Sequence
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Animals
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Anti-Infective Agents
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chemical synthesis
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pharmacology
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Antimicrobial Cationic Peptides
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chemical synthesis
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chemistry
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pharmacology
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Molecular Sequence Data
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Mytilus
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chemistry
8.Construction of an evaluation index system for the core competence of chemotherapy specialist nurses
Yang YANG ; Mei XUE ; ge Yuan DONG ; qi Zhen LU ; yan Hai LU ; ju Xiao ZHANG ; li Ling GU ; Jing SHI ; Yang WANG
Chinese Journal of Nursing 2017;52(10):1226-1231
Objective To construct an evaluation index system for the core competence of tumor chemotherapy specialist nurses in China and to provide standards for training of tumor chemotherapy specialist nurses.Methods Through literature research and expert meeting,the first draft of index system was established,and consultation questionnaire was compiled.Two rounds of consultation were conducted with 23 experts in China using Delphi method.AHP method was used to determine the weights of indicators at all levels.Results The effective rates of 2 rounds of expert consultation were 100% and 73.91%,respectively.In the 2 rounds of consultation,the expert authority coefficients were 0.855 and 0.867,respectively.The coordination coefficient of first and second level indexes and expert opinions were 0.693 and 0.371 (P<0.001),respectively.The final evaluation index system for the core competence of tumor chemotherapy specialist nurses included 5 first-level indexes (positive professional attitude,sufficient clinical practice ability,strong educational counseling ability,basic clinical research and evidencebased practice ability,and basic management ability),16 second-level indexes,and 59 third-level indexes.Conclusion The evaluation index system for chemotherapy specialist nurses based on core competence determined by expert consultation is scientific and reasonable,and can provide a powerful basis for training qualified chemotherapy specialist nurses.
9.The effects of stem cell factor on proliferation, transmigration, capillary tube formation of endothelial cells and on the chemotaxis of CD133(+) cells.
Xin YAN ; Bin LIU ; Shi-hong LU ; Mei-li GE ; Xing-xin LI ; Yi-zhou ZHENG
Chinese Journal of Hematology 2011;32(5):326-330
OBJECTIVETo explore the effects of stem cell factor (SCF) on proliferation, transmigration, capillary tube formation of human umbilical vein endothelial cells (HUVEC) and on the chemotaxis of CD133(+) cells.
METHODSIn the presence of blank control, SCF, vascular endothelial growth factor (VEGF), anti-human SCF (anti-SCF) or human IgG, the difference in proliferation capacity of HUVEC was analyzed by MTT and CCK-8 methods, and wound scratch assay and three-dimensional in vitro Matrigel assay were used for transmigration and capillary tube formation of HUVEC, respectively. In addition, the chemotaxis of CD133(+) cells sorted from human umbilical cord blood by flow cytometry was investigated by Transwell migration assay.
RESULTSSCF didn't improve the proliferative capacity of HUVEC, but significantly enhanced the transmigration capacity, and increased capillary tube formation in a dose-dependent manner. The number of intact tubules [(30.0 ± 3.4)/10(5) HUVEC] formed by HUVECs in the presence of the optimal concentration of SCF (100 ng/ml) was remarkably higher than that in blank control group [(5.0 ± 2.6)/10(5) HUVEC, P < 0.01]. SCF also significantly induced a chemotactic response of CD133(+) cells, the transmembrane migration cell number into Transwell lower chamber was significantly higher in SCF group [(118.0 ± 6.5)/10(4) CD133(+) cells] than in blank control group [(47.0 ± 4.7)/10(4) CD133(+) cells, P < 0.01 ].
CONCLUSIONSSCF significantly promotes the transmigration and capillary tube formation of HUVEC, and induces a chemotactic response of CD133(+) cells. SCF/c-kit signaling possibly plays a critical role in regulating angiogenesis of vascular endothelial cells and vasculogenesis of endothelial progenitor cells.
Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Neovascularization, Physiologic ; drug effects ; Sincalide ; metabolism ; Stem Cell Factor ; pharmacology
10.Study on the mechanism of acupoint sticking therapy with Chuan fuling for treatment of asthma model rats.
Xi-xiong XIANG ; Jing-ying ZHANG ; Yin-mei SI ; Chang-yu LIU ; Yan-fang LU ; Xiao-ying LIU ; Wang-xi CAI ; Han-ge FENG
Chinese Acupuncture & Moxibustion 2009;29(9):739-743
OBJECTIVETo investigate the mechanism of the acupoint sticking therapy with Chuanfuling for preventing and treating asthma.
METHODSThirty male SD rats were randomly divided into a control group (normal saline, p.i. +no acupoint sticking+ normal saline, spray inhalation), model group (normal saline with ovalbumin, p.i. +no acupoint sticking+ normal saline with ovalbumin, spray inhalation), and acupoint sticking group (normal saline with ovalbumin, p.i. +acupoint sticking with Chuan fuling+normal saline with ovalbumin, spray inhalation), 10 rats in each group. The incubation period of nodding breath, symptom of asthmatic attack, expression level of interleukin-4 mRNA (IL-4 mRNA) and interferon-gamma mRNA (IF-gamma mRNA), as well as pathological changes on the middle leaf of right lung, were observed in each group.
RESULTS(1) Comparing with the control group, the model group was showed that the expression level of IL-4 mRNA in the peripheral blood cells (PBMC) was increased, while hyperemia, edema and eosinocyte (EOS) invasion of lung tissue was more serious (P < 0.01). (2) Comparing with the model group, the acupoint sticking group was showed that the expression level of IL-4 mRNA in PBMC was decreased, the incubation period of nodding breath was prolonged for induced asthma on the fifth and seventh time with lower frequency, while in the lung tissue EOS invasion was reduced (P < 0.05), but there were no significant changes on the hyperemia and edema (P > 0.05).
CONCLUSIONAcupoint sticking for treating asthma of model rats with Chuanfuling can inhibit the expression level of IL-4 mRNA in PBMC, and the release of the inflammatory mediator and cytokine from the EOS to the air passage, in order to reduce the injury of epithelial layer and high reaction on the air passage.
Acupuncture Points ; Animals ; Asthma ; drug therapy ; genetics ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Gene Expression ; drug effects ; Humans ; Interleukin-4 ; genetics ; immunology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley