Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.

Objective To assess the effects of thalidomide on VEGF mRNA level in human breast cancer cell lines. Methods Breast cancer cells were treated with thalidomide for 24～72h, RT-PCR was uced to detect the level of VEGF mRNA expression. Results When the concentration of drug is 50μg/L,thalidomide strongly inhibited the level of VEGF mRNA expression. Conclusion Within certain concentration range of thalidomide can inhibit the level of VEGF mRNA expression in breast cancer cell.

Objective To examine the clinical effect of simultaneous intervention for heart and lung on chronic obstructive pulmonary disease (COPD) of stationary phase combined with stable angina pectoris with Qi deficiencyblood stasis-phlegm blockade syndrome.Methods Ninety-six COPD stationary phase combined with stable angina pectoris patients with Qi deficiency-blood stasis-phlegm blockade syndrome were randomized into control group,Juhong [Exocarpium Citri Rubrum] tablet group,the Tongxinluo (通心络) group and the Ju&Tong group,24 cases in each group.The control group was given western medicine routine therapy.In addition to the treatment of the control group,Juhong tablet 3.6 g was given to the Juhong tablet group orally,twice each day;Tongxinluo capsule 1.04 g was given to the Tongxinluo group orally,three times each day;Juhong tablet and Tongxinluo capsule were given to the Ju&Tong group.Each group was treated for 8 weeks.The following intems were compared before and after treatment including the scores of cough,cough up phlegm,dyspnea and St.George's Respiratory Questionnaire (SGRQ),anginal attacks,durante dolors,nitroglycerin consumption,pulmonary function [including forced expiratory volume in one second (FEV1) and forced vital capacity (FVC)],as well as the levels of serum C reactive protein (CRP),interleukin 1β (IL-1β) and interleukin 10 (IL-10).Results After treatment,the scores of cough,cough up phlegm,dyspnea and SGRQ decreased in the Juhong tablet group,the Tongxinluo group and the Ju&Tong group.FEV1 and FVC increased.Anginal attacks,durante dolors,nitroglycerin consumption,as well as the levels of serum CRP,IL-1 βand IL-10 decreased.Moreover,the effect of certain indexes in the Ju&Tong group was superior to those in the Juhong tablet group and the Tongxinluo group (P ＜ 0.05 or P ＜ 0.01).Conclusion Simultaneous intervention for heart and lung might improve clinical symptoms and pulmonary function of COPD stationary phase combined with stable angina pectoris with Qi deficiency-blood stasis-phlegm blockade syndrome patients.Inhibiting chronic persistent inflammation might be one of the important mechanisms.

Objective:To analyze the effects of DHT on the proliferation and migration of endothelial progenitor cells (EPCs) and the role of RhoA/ROCK pathway in this process.Methods:Early EPCs were isolated from peripheral blood of healthy adults, and cultured in serum-free EBM-2 medium for 24 h before incubation with various concentrations of DHT (1, 10, and 100 nmol/L). EPCs proliferative and migrative capacities were measured. The adherent cells were collected and randomLy divided into: control group, DHT group, C3 exoenzyme+DHT, Y-27632+DHT group. EPCs proliferation and migration were assayed by MTT assay and modified Boyden chamber assay respectively.Results:DHT significantly increased the proliferation and migration ability of EPCs in a dose- and time-dependent manner, maximum at 10 nmol/L, 24 h ( P<0.05). C3 exoenzyme [(0.22±0.02) vs (0.26±0.05), P>0.05] and Y-27632 [(0.21±0.04) vs (0.26±0.05), P>0.05] can attenuate the proliferative capacities of EPCs induced by DHT compared with the DHT group, but there was no statistical significance. The influence of DHT on EPCs migrative capacities can be abolished by C3 exoenzyme [(35.26±4.27) vs (46.92±5.46), P<0.05] and Y-27632 [(33.61±5.33) vs (46.92±5.46), P<0.01]. C3 exoenzyme [(116.75±7.42) vs (156.80± 21.74), P<0.05] and Y-27632 [(121.73±5.33) vs (156.80 ±21.74), P<0.01] could noticeably attenuate DHT-induced EPCs secretion of VEGF respectively. Conclusions:DHT can modulate EPCs proliferation, migration and the RhoA/ROCK pathway plays an important role in this process.

OBJECTIVETo study the clinical and genetic characteristics of Pallister-Killian syndrome and improve the diagnosis for this rare chromosomal disease.

METHODSStandard G-banding was carried out for the patient and his parents. Single nucleotide polymorphism array (SNP array) for copy number detection was applied to identify chromosome microdeletion or microduplication. Interphase fluorescence in situ hybridization (FISH) and cytogenetic analyses of fibroblast cells were performed based on the Results of array.

RESULTSThe patient's G-banded karyotype has turned out to be 46,XY, whilst his parents were both normal. A duplication of the whole short arm of chromosome 12 was detected by SNP array in the child. The result of interphase FISH performed on interphase chromosomes derived from peripheral blood cells was nucish (RP11-104 b5, a19 RP11-956) × 4 [19/100］, whilst the karyotype of fibroblast cells was 47,XY,+i(12) (p10 [44]/46, XY[56].

CONCLUSIONBy combining with clinical characteristics, SNP array, skin fibroblasts karyotype analysis and FISH can diagnose Pallister-Killian syndrome effectively.

Chromosome Banding ; Chromosome Disorders ; diagnosis ; genetics ; Chromosomes, Human, Pair 12 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo apply single nucleotide polymorphism microarray (SNP array) for the detection of genome-wide copy number variations(CNVs) in fetuses with malformations and women with an adverse reproductive history, and to explore the correlation of rare CNVs with the clinical manifestations.

METHODSAmniotic fluid and umbilical cord blood samples were collected from 314 women with singleton pregnancy. SNP array was performed on samples where chromosomal abnormalities were excluded after G-banding analysis.

RESULTSPathological CNVs were detected in 8.91% (28/314) of all samples, which included 11 duplications, 9 deletions, 4 loss of heterozygosity (LOH), and 4 conjoined deletions and duplications. The sizes of duplications and deletions were between 0.47 Mb and 16.7 Mb, and between 0.16 Mb and 13.3 Mb, respectively. Fifteen CNVs were mapped to the regions of microdeletion or microduplication syndromes or regions associated with clinical manifestations, while the remainder 13 were considered benign or variant of uncertain significance.

CONCLUSIONA proportion of fetuses with malformations and women with an adverse reproductive history may be attributed to CNVs, half of which are mapped with to the regions of well known syndromes. SNP array may facilitate discovery of new syndromes and provide a basis for genetic counseling and prenatal diagnosis.

Adult ; Chromosome Aberrations ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; DNA Copy Number Variations ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Polymorphism, Single Nucleotide ; Pregnancy ; Pregnancy Complications ; diagnosis ; genetics ; Prenatal Diagnosis ; Reproductive History ; Young Adult