3.Study on the amplifying and typing of low copy number DNA
Chinese Journal of Forensic Medicine 2009;24(6):383-385,403
Objective To study the influence on STR typing for low copy number(LCN)DNA using different methods of amplifcation and detection.Methods Control DNA 9947A was diluted and then amplifled with Identifiler~(TM) or DNATyper15~(TM).The heat cycles were set to 28 or 28 add 6.Each template wag amplified three times in parallel,and then the amplified products or the product mixture of three amplifictions were deteced with 310 or 3130 Genetic Analyzer.Results The success rate of STR typing with the method 28+6 cycles was higher than that of 28 cycles.There are no correlation between the allele imbalance or allele dropout and STR locus.With the reduction in the amount of template DNA,allele imbalance and dropout gradually increased,and the allele dropout was more common than allele imbalance when the amount of template DNA Wag very small.When the product mixture of three amplifictions were deteced.the occurrence of allele imbalance and dropout reduce obviously.Conclusion The success rate of STR typing of LCN DNA can be obviously increased by detecting the product mixture of three amplifictions in parallel combined with the 28+6 heat cycle condition.
4.Preparation of internal quality control substance of real-time PCR to analyze UU-DNA and the design quality control programs
Yanmei GE ; Di ZHANG ; Suyi FAN ; Hang YUAN ; Yuan MAO
International Journal of Laboratory Medicine 2016;37(8):1070-1071,1074
Objective Developing an internal quality control substance of Ureaplasma urealyticum(UU)‐DNA for real‐time PCR to establish an internal quality control system and preliminary evaluation its clinical value .Methods Internal quality control sub‐stance was prepared by mixing samples which Ct value were 24-25(positive sample) and 32 -33(weak positive sample) ,respec‐tively .At the same time ,selecting samples that test results were negative as negative control .The target value ,standard deviation (s) and coefficient of variation(CV) of internal quality control substance were defined by“instant method”for the first 20 runs and Levey‐Jennings quality control(QC) chart after the first 20 runs .Using the“Westdard” multi‐rule quality control methods to ana‐lyze the detection results .Exporting OPSPecs chart by quality control rules in Unity Real Time (URT ) system and setting up new quality control rules according with OPSPecs chart .Results 131 times of the detection of quality control substance were performed totally .The first 20 runs were defined by“instant method”and later 111 runs were defined by Levey‐Jennings QC chart ,the results were stable of quality control substance and reasonable quality control rules .Conclusion Preparing of internal quality control sub‐stance of UU‐DNA used in real‐time PCR might be easy and stable .So ,the internal quality control substance of UU‐DNA could be worthy for practical application in this PCR laboratory .Design internal quality control rules based OPSPecs chart in molecular de‐tection is very simple and practical .
5.Efficacy Observation for Treating Ankylosing Spondylitis by Chinese Herbs and Recombinant Hu- man Tumor Necrosis Factor Receptor II-Antibody Fusion Protein.
Wei LIU ; Di ZHANG ; Yuan-hao WU ; Hui-jun YANG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(6):663-667
OBJECTIVETo observe the clinical effect of Chinese medical (CM) syndrome differentiation based Chinese herbs and recombinant human tumor necrosis factor receptor II-antibody fusion protein (etanercept) for treating ankylosing spondylitis (AS) patients.
METHODSTotally 35 AS patients were treated with syndrome differentiation based Chinese herbs and etanercept. Reinforcing Shen and strengthening Du channel, activating meridians to stop pain was principle used in syndrome differentiation based treatment. Etanercept was subcutaneously injected, 25 mg each time; twice per week for the first three months and once a week for the latter three months. The clinical efficacy was evaluated after 3 and 6 months of treatment. Meanwhile, ASAS20 and ASAS50 standards arriving rates were also observed. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI), visual analog score (VAS) for spine pain, VAS for night pain, patient global assessment (PGA), VAS for physician global assessment, CM syndrome score, finger-ground distance, thoracic activity, tragus-wall distance, lumbar scoliosis, cervical rotation, Schober improved test, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were observed before treatment, 3 and 6 months after treatment.
RESULTSCompared with before treatment, BASDAI, BASFI, VAS for spine pain, night pain, physician global assessment, PGA, CM syndrome score, finger-ground distance, thoracic activity, tragus-wall distance, lumbar scoliosis, Schober improved test, ESR, and CRP all decreased after 3 and 6 months of treatment, with statistical difference (P < 0.05). Cervical rotation also decreased after 6 months of treatment, with statistical difference (P < 0.05). Compared with 3 months of treatment, total effective rate of CM syndrome, ASAS20 and ASAS50 standards arriving rates increased after 6 months of treatment, with statistical difference (P < 0.05). There were statistical differences in all indices mentioned above between after 3 months of treatment and after 6 months of treatment (P < 0.05).
CONCLUSIONSyndrome differentiation based Chinese herbs combined etanercept could alleviate inflammatory reaction favorably, control the progression of active AS, and improve joint functions.
Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Etanercept ; therapeutic use ; Humans ; Pain ; prevention & control ; Pain Management ; Spondylitis, Ankylosing ; drug therapy ; Treatment Outcome
7.Analysis of clinical characteristics and JAK2V617F mutation of Tibetan people living at high altitudes with polycythemia.
Di-jiao TANG ; Yuan-xin YE ; Jun LI
Chinese Journal of Hematology 2012;33(11):960-962
Adult
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Altitude
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Asian Continental Ancestry Group
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genetics
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Female
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Humans
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Janus Kinase 2
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genetics
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Male
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Middle Aged
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Mutation
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Polycythemia
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diagnosis
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ethnology
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genetics
8.Inhibitory effects of andrographis paniculata and silybum marianum on the efflux pump of MRSA 41577
Linhui YUAN ; Di WANG ; Kunpeng XIE ; Mingjie XIE
Chinese Journal of Biochemical Pharmaceutics 2016;36(11):27-29
Objective To study the inhibitory effects of andrographis paniculata and silybum marianum on the efflux system of MRSA 41577. Methods Inhibitory effects of andrographis paniculata and silybum marianum on efflux system of MRSA 41577 was evaluated using fluorescence spectrophotometry.PCR was applied to detect the norA efflux gene.By RT-PCR method for detection of andrographis paniculata and silybum marianum influence of the expression of norA efflux gene.Results Andrographis paniculata and silybum marianum significantly increased the accumulation of ciprofloxacin in MRSA 41577 in a time-dependent manner.At 12 minute, andrographis paniculata and silybum marianum respectively increased ciprofloxacin in MRSA41577 by 49% and 76%( P <0.05 ) , which is superior to that of reserpine. Further mechanism studies indicated that andrographis paniculata and silybum marianumcould reduce the expression of norA in MRSA 41577.After incubated with andrographis paniculata and silybum marianum for 16 h, the relative expression of norA of MRSA41577 was respectively reduced by 35% and 42% ( P <0.05 ). Conclusion Andrographis paniculata and silybum marianumcould inhibit MRSA efflux system through reducing pathogen ’s expression of norA and NorA protein.
9.Influence of subculture cycle on tube plantlet growth of Dendrobium huoshanense and medium composition
Shuhua JIA ; Di WANG ; Yuan GAO ; Yongping CAI ; Yi LIN
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective To optimize the best subculture cycle through studying the influence of subculture time on tube plantlet growth of Dendrobium huoshanense and the change of medium composition.Methods Height,tiller,fresh weight,and chlorophyll content of the tube plantlet and pH value,water content,and sugar content of the medium were measured after different cycles of the subculture,the cost of culture medium for subculture was calculated as well.Results The height of the tube plantlet increase 282.86%,the tiller increase by 3.5 times,fresh weight reaches its maximum,chlorophyll content of the tube plantlet almost reaches its maximum after 40 d subculture;while water content and sugar content of the medium are decreased to the lowest point,pH value of medium(
10.Comparison of three different flow cytometers in the clinical assessment of lymphocyte subsets
Weiwei WANG ; Di XI ; Xiangliang YUAN ; Lisong SHEN
Chinese Journal of Laboratory Medicine 2016;39(5):361-365
Objective To evaluate the consistency and accuracy among 3 brands of flow cytometers (BriCyte E6,BD FACSCanto Ⅱ and Beckman Coulter FC 500) in the detection of lymphocyte subsets.Methods According to the methodology,the BriCyte E6 was compared with 2 flow cytometers commonly used in clinical detection.Seventy-three cases (40 male and 33 female) of anticoagulation peripheral blood specimens were collected in the clinical laborartory department of Xinhua Hospital in July 2015 and the percentage (%) and absolute number (#) of the lymphocyte subsets were detected by 3 different flow cytometers within samples collected 4 h.Results There were good consistency among the 3 flow cytometers (R2 >0.95,R2 from 0.969 5 to 0.992 4) in the detection of lymphocyte subsets percentage,so did in the detection of absolute number (R2 > 0.95,R2 from 0.969 1 to 0.993 3).As to the precision evaluation,in the detectionof CD8%,T#,CD4+ T# and CD8+ T#,BriCyte E6 achieved a low CV% compared with FACSCanto Ⅱ and FC 500 (Friedman statistics are 16.720,11.840,15.760 and 15.430,P =0.000 2,0.027,0.000 4,0.000 4,respectively).In the detection of T%,CD4%,NK%,B%,NK#,B#,there was no significant difference among the 3 flow cytometers (Friedman statistics are 4.242,3.916,0.852,2.595,1.835 and 0.578,P =0.119 9,0.141 2,0.653 2,0.273 3,0.399 6,0.749 0,respectively).Conclusions The 3 flow cytometers have a good consistency in the detection of lymphocyte subsets.BriCyte E6 may be an alternative or complement of existing flow cytometers.