Objective To study the influence of EDTA Dependent Leukoagglutination on the white blood cell counting result by automated blood analyzer,the possible reason of WBC aggregation caused by EDTA anticoagulant and the way to correct its influence. Methods Statistics and analysis,from October 2014 to October 2016,345600 cases of outpatient and inpatient blood routine specimens,EDTA induced leukocyte accumulation,leaded to the number of cases of false white blood cells. Results It revealed that the incidence was 0.002%(7 cases among 345,600) of which had EDTA induced Leukoagglutination,27 cases(0.008%)had WBC clumping caused by other reasons. These 34 samples had spuriously low WBC counts due to WBC clumping compared to manual differential results(P<0.01). The Leukoagglutination caused by EDTA could not be corrected by incubating samples in 37℃ water bath for 30 mints or by replacing the EDTA by sodium citrate. The leukoagglutination caused by EDTA could be partially be corrected by diluting the sample with instruments diluent at a dilution of 1:1 and by adding kanamycin 20 mg/ml to samples,but still influence the white blood cell counting resulted by automated blood analyzer(P<0.001). Conclusion EDTA can induce WBC aggregation,and thus spuriously low WBC counts,our study shows that the incidence is 0.002%. Examination of a well prepared peripheral blood smear is mandatory for every case of leukoagglutination.

Dendrobium candidum is a traditional medicinal plant critically endangered in China. Three molecular markers, namely, inter-simple sequence repeat (ISSR), sequence-related amplified polymorphism (SRAP), and start codon-targeted (SCoT), were used to analyze the genetic diversity of 40 varieties of D. candidum. Ten primers were screened for ISSR, SRAP, and SCoT PCR reactions individually, and the polymorphic ratios were 84.6%, 76.5%, and 90.3%, respectively. Analysis using the three markers showed that genetic variations were abundant among different sources of D. candidum but few among different Dendrobium species. Some cultivars also showed similarities to the wild species. Results indicated the abundant genetic diversity of D. candidum cultivars among different areas. ISSR, SRAP, and SCoT markers exhibited similar efficiencies for fingerprinting genotypes; however, SCoT was the most effective in fingerprinting D. candidum varieties.

目的 研究干扰素调节因子8(interferon regulatory factor 8,IRF-8)在骨髓增生异常综合征伴原始细胞增多(Myelodysplastic Syndromes with Excess Blasts,MDS-EB)亚型中是否存在表达异常,并分析其与MDS-EB的相关性.方法 收集三组共30例实验标本.其中20例为MDS-EB病例标本,分为MDS-EB1组和MDS-EB2组.10例为健康体检的健康对照组.30例标本均采集自受试者外周静脉血,采集后立即进行外周血总RNA提取,并经逆转录PCR后,以实时荧光定量PCR检测各标本的IRF-8 mRNA表达水平.各组平均表达量以RQ值(x±s)表示.结果 三组样本实时荧光定量PCR的平均RQ值:MDS-EB1组(0.59±0.08),MDS-EB2组(0.26±0.10),健康对照组(1.20±0.26),三组表达水平存在显著差异(P<0.01).MDS-EB组的IRF-8表达水平均显著低于健康对照组(P<0.01),MDS-EB2组内IRF-8表达水平低于MDS-EB1组(P<0.05).结论 IRF-8在MDS-EB中表达水平降低,表明IRF-8在MDS-EB中表达受抑,组间统计结果 提示IRF-8的表达可能与疾病的恶性程度及进展相关.

Objective] To investigate the effects of inhibiting pancreatic cancer of Cinobufotalin by bioluminescent imaging(BLI) in the BxPC3-luc2 cell xenograft nude mice.[Method] Twenty-four nude mice were subcutaneously injected BxPC3-luc2 pancreatic cancer cells. After seven days, tumor-bearing mice were divided into three groups:blank group, Cinobufotalin group(6g·kg-1, Cinobufotalin) and positive drug group(2mg·kg-1 DDP), eight mice for each group and continuous intraperitoneal injection of 14d. The tumor growth of pancreatic cancer in nude mice was detected by using the card scale and living creature luminescence imaging instrument at 7th and 14th days, and the animals were sacrificed at 14th day, and the solid tumor was weighed. [Result] Tumor bearing nude mice with injection of Cinobufotalin had no significant changes in body weight compared with the control group, while tumor weight and volume were significantly reduced; Tumor photon was also significantly reduced in nude mice treated with Cinobufotalin at after administration of 7th and 14th days, the inhibition rates of tumor photon were 30.11%and 63.81%respectively. But 6g·kg-1 Cinobufotalin was weaker than 2mg·kg-1 DDP. The effect of Cinobufotalin did not change significantly for body weight in mice, had certain advantages. [Conclusion] Cinobufotalin has a good inhibition of BxPC3 pancreatic cancer in nude mice.

Objective: To analyze the genetic diversity of Dendrobium candidums by start condon targeted polymorphism(SCoT) markers.Methods: the molecular marker technique of SCoT and unweighted pair-group method with arithmetic means(UPGMA) were used to study the genetic diversity of 20 artificial cultivar samples.Results: 20 primers were screened from 56 SCoT primers and then used to amplify the genome DNAs of the accessions.Totally 226 SCoT loci were produced,including 204 polymorphic loci,with a polymorphism rate of 90.3%.Conclusion: It was revealed that there were abundant genetic variations among different sources D.candidum,the genetic diversity of artificial cultivation D.candidum was rich which was sold on the market at present,and the efficiency of SCOT markers was relatively the same in fingerprinting of genotypes of D.candidum varieties.

The concentration gradient was used to optimize reaction system for sequence-related amplified polymorphism(SRAP) of Bletilla striata.The suitable concentration of template DNA,primer,Mg2+,dNTPs,rTaq DNA polymerase and annealing temperature were investigated through single factor test.The SRAP-PCR amplifying system was optimized which came out total 20 μL reaction system containing: 30ng DNA template,2.5 mmol/L Mg2+,0.25 μmol/L dNTPs,20 μmol/L primers,2.0U Taq DNA polymerase,the most suitable PCR amplification procedure was pre-denaturing at 94 ℃ for 5 min;5 cycles of denaturing at 94 ℃ for 1 min,annealing at 36 ℃ for 1 min and extending at 72 ℃ for 1 min,then 35 cycles of denaturing at 94 ℃ for 1 min,annealing at 48 ℃ for 1 min and extending at 72 ℃ for 1 min,extending at 72 ℃ for 10 min finally.The optimized system laid the groundwork for SRAP molecular analysis and further research on Bletilla striata.

Objective In order to optimize the reaction system of start condon targeted(SCoT) polymorphism of Dendrobium officinale,and to lay the foundation for analysis on genetic diversity of D.officinale.Methods The fresh leaves of D.officinale were used and the effects of template DNA,primer,Taq DNA polymerase,dNTPs,and annealing temperature on SCoT-PCR amplification were determined through single factor test.Results The SCoT-marked 20 μL optimized reaction system of D.officinale included 20 ng template DNA,30 μmol/L primer,1.0 U Taq DNA polymerase,and 100 μmol/L dNTP.The suitable PCR amplification procedure was one cycle of pre-denaturing at 94 ℃ for 4 min,36 cycles of denaturing at 95 ℃ for 50 s,annealing at 56.1 ℃ for 40 s,and extending at 72 ℃ for 2 min,extending at 72 ℃ for 5 min,and finally,being kept at 4 ℃.PCR amplification results by 1.5% agarose gel electrophoresis showed that amplification products were mainly in the range of 600-2 000 bp.Conclusion The established SCoT-PCR reaction system could provide the reference for germplasm genetic diversity analysis and molecular marker-assisted breeding of D.officinale.