OBJECTIVETo evaluate value of spectral karyotyping (SKY) in the detection of chromosomal abnormalities.

METHODSA total of 17 metaphase chromosome samples were investigated by SKY, including 10 normal and 5 balanced translocation samples of peripheral blood lymphocytes, one der(Y) sample of peripheral blood lymphocytes and one marker chromosome sample of amniotic fluid cells. The results were compared with those of G-banding diagnosis.

RESULTSTen normal and 5 balanced translocation samples were diagnosed successfully by SKY in accordance with the results of G-banding; furthermore, SKY analysis revealed that the der(Y) fragment originated from p-arm of chromosome 21 while the marker chromosome originated from chromosome 5.

CONCLUSIONSKY is a very sensitive and specific whole genome analysis tool for chromosomal abnormality diagnosis, and exceedingly valuable in the diagnosis on complex chromosomal abnormalities that can not be determined by G-banding.

Chromosome Aberrations ; Chromosome Banding ; methods ; Chromosome Disorders ; diagnosis ; genetics ; Female ; Humans ; Pregnancy ; Prenatal Diagnosis ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Spectral Karyotyping ; methods

Objective To build a rat model of intrauterine adhesion (IUA) by whole layer removal of endometrium in order to provide a suitable animal model for the treatment and experimental study of endometrial lesions and IUA.Methods According to different scopes of endometrial excision,105 female SD rats were divided into group A (2.5 cm × 0.5 cm),group B (2.5 cm × 0.25 cm) and group C (sham operation group) with 35 rats in each group.Uterine tissues of rats were collected 3,7,15 and 30 days after surgery.The ratio of uterine cavity narrow and congestion was recorded,the number of glands in the endometrium and the ratio of endometrial fibrosis area was counted,microvascular density (MVD) was measured,and pregnancy rate of rats in each group were observed to compare the two excision methods.Results The ratio of fibrosis area in group A and group B (0.892 ± 0.068,0.562 ± 0.027) were higher than that in group C (0.374 ± 0.074).Uterus patency scores in group A and group B (1.87 ± 0.16,1.20 ± 0.17) were both significantly higher than that in group C (0.68 ±0.098).The differences were statistically significant (P ＜ 0.001).The numbers of glands in the endometrium 30 days after surgery in group A,B and C were 7.35 ± 2.19,14.5 ± 2.43 and 15.56 ± 2.63,respectively,significantly less in group A than that in group B and C (both P＜0.001),while there was no statistical difference between group B and C (P =0.054).The MVD in group A,B and C were 16.06 ± 2.11,24.6 ± 2.34 and 26.37 ± 5.09,respectively,significantly less in group A than that in group B and C (both P＜0.001),while there was no statistical differences between group B and C (P=0.303).The rates of pregnancy in group A,B and C were 20％,33.3％ and 100％,respectively,obviously lower in group A than that in group B and C (P＜0.05).Conclusions The method of whole layer removal of endometriun had a high success rate in establishing a stable and effective rat model of IUA.

Squalene synthase of Alisma orientale catalyzes farnesyl diphosphate (FPP) to form squalene, which is the key regulatory enzyme of the carbon source flow to protostane triterpenes biosynthesis. For further research on the function and expression of AoSS gene, the open reading frame (ORF) of squalene synthase gene (accession no. JX866770) from A. orientale was subcloned into a prokaryotic expression vector pCzn1 and induced the expression of AoSS gene in Escherichia coli BL21(Roseta). The fusion protein was mainly in the form of inclusion bodies and purified to obtain high purity protein. By verifying its functionality through vitro enzymatic reaction, the results showed that the catalytic protein had the catalytic activity of FPP into squalene. In order to research the expression of AoSS in A. orientale, the purified protein was used to immunized rabbits to prepare polyclonal antibody which was then purified, the titer of the antibody was greater than 1∶51 200 by ELISA detection, and displayed good specificity by Western blotting. The prepared antibody was used for immunoassay of AoSS in different organs of A. orientale, and the results showed that the AoSS expression level was the highest in tubers, followed by leaves, and lowest in root. Successful construction of prokaryotic expression vector, validation of gene functions and establishment of rapid immunoassay lay the foundation for further researches on the function and regulation of AoSS gene, and also provide scientific basis on the application of the protostane triterpenes of A. orientale in the field of synthetic biology.

Objective Throush identify biochemical characteristics and virulence factors of 2 strains suspected Yersinia pestis(Y.pestis)isolated from the dead Marmota himalayana(M.himalayana)to confirm the nature epidemic focus in Dege County,Sichuan Province.Methods Y.pestis was analyzed by specific staining and shape,culturing characteristics,splitting-test by bacteriophage,test of biochemical characteristics and glycolysis ability,virulence factors,virulence,nutritional requirement,plasmid,genetic test and genetic type. Results The tested strains were Gram staining bacilus.The main biochemical characteristics were Arabinose(+)、 Rhamnose(-),Maltose(+),Melibiose(-),Glycerol(+),Denitrification(+).The virulence factors with FI+.VW+, Pgm+,Pst I+;and with the common 6.0×106,45.0×106,65.0×106 plasmids,also with the virulence-relative plasmid gene.Both their absolutely lethal dose(LD100)in mice were 50 bacteria.The nutritional requirement appeared which were depended on Phenylalanine and Methionine.With the Genomovar 5 genotype characteristics of M.himalayana plague foci of Qinghai-Tibet plateau.The difference between tested strains and Yersinia pseudotubercuosis on the 3 different culture medium was obvious.The tested strains had a Y.pestis' specific 3a fragment,Pst I and FI-Ag,at 22 ℃,the strains could be split by bacteriophage completely.Conclusions According to the diagnostic criteria of plague in China,the 2 suspected strains isolated from Dege County,Sichuan Province ale confirmed as Y.pestis.both with powerful virulenceand with the characteristics of the Y.pestis of M.himahtyana in Qinghai-Tibet plateau plague natural focus.