1.Diagnosis of fetal chromosomal aneuploidy by free fetal DNA in maternal plasma in the first trimester
Ying CHANG ; Xu CHEN ; Hongyan CUI ; Chenchun REN
Chinese Journal of Perinatal Medicine 2014;17(1):14-18
Objective To assess the clinical value of fetal chromosomal aneuploidy diagnosed by free fetal DNA in maternal plasma in 11-13+6 gestational weeks.Methods A total of 2 650 pregnant women who had prenatal care in Tianjin Center Hospital of Obstetrics and Gynecology from January 1,2010 to December 31,2010 were included.Each of them had an ultrasound scan to measure fetal nuchal translucency thickness.Maternal serum free β-human chorionic gonadotropin and pregnancy-associated plasma protein A test was performed as part of screening for chromosomal abnormalities.Results of ultrasound and maternal plasma biochemical analysis were entered into the database,and converted into multiple of median (MoM) by factors such as maternal age,weight,ethnicity,smoking history and mode of conception.The cutoff value was 1 ∶ 270.Meanwhile,20 cases had cell free fetal DNA (cffDNA) test and the ratio of the single nucleotide polymorphism on two alleles of plancenta-specific 4 (PLAC4) were measured in 16 cases.T-test,rank sum test,MannWhitney U test and Chi-square test were used as statistical methods.Results (1) A total of 74 cases were judged as high-risk,among which 35 cases underwent transabdominal chorionic villus sampling (18 cases had cffDNA test),37 cases underwent amniocentesis at the week of 20,and two cases of Rh negative did not receive the invasive examination.Totally 20 cases,including two Rh negative cases,had the cell-free fetal DNA test.(2) By cffDNA test of maternal plasma,two cases of 21 trisomy,one case of 18 trisomy,two cases of 45,XO and one case of balanced translocation were diagnosed.(3) In the two cases of 21 trisomy,maternal plasma G/A ratio ofPL4C4 RNA-single nucleotide polymorphism alleles was 1.00 (0.98,1.02) ; in 14 pregnancies with normal chromosome,the ratio was 1.055 (1.02,1.13,Z=3.5).There was no significant difference (P=0.066).Conclusion Diagnosing of fetal chromosomal aneuploidy by cffDNA in maternal plasma is feasible and noninvasive with high negative predictive value,and can be used in Rh-negative pregnant women for prenatal screening and diagnosis.
2.Study on several ultrasound markers combined maternal serum biochemical markers to screen fetal chromosomal aneuploidy at 11 to 13+6 weeks of gestation
Xu CHEN ; Ying CHANG ; Hongyan CUI ; Chenchun REN ; Bingying YU
Chinese Journal of Obstetrics and Gynecology 2013;48(11):815-818
Objective To evaluate the efficiency of combined screening for chromosomal abnormalities in the first trimester and the ultrasound characteristics of these fetuses.Methods Retrospective study for 5000 singleton pregnancies by combined screening of trisomies 21,18,13 and Turner syndrome.Risk algorithms were developed for calculation of patient-specific risks for each of the three trisomies based on maternal age,fetal nuchal translucency,free β human chorionic gonadotropin and serum pregnancy associated plasma protein A at 11 to 13 +6 weeks of pregnant.The value of nuchal translucency (NT) and β-hCG and pregnancy-associated plasma protein A (PAPP-A) level were inputted computer,and calculate the risk value (≥ 1 ∶ 270) by automatic analysis software.Two hundred and four cases with high risk were performed transabdominal chorionic villus biopsy to detect the fetal chromosomal karyotypes.Meanwhile,other ultrasonic characteristics of fetal were elevated.Results (1) Five thousand cases of pregnant women were detected,including 4983 normal cases,62 cases were induced labor for a variety of reasons in the second trimester,including 40 cases with normal karyotype but with congenital heart disease,17 cases of chromosome abnormalities (9 cases trisomy 21,2 cases trisomy 18,1 cases trisomy 13,4 cases 45X),2 cases spina bifida,2 cases digestive tract obstruction,1 cases giant bladder.One case with low risk of fetal chromosomal abnormalities in combined screening,but high risk of age (maternal age were over 40 years old),it was 21 trisomy syndrome after the prenatal diagnosis.(2) Five cases of nasal bone loss in 9 cases of trisomy 21 (5/9),5 cases with three tricuspid regurgitation (5/9),4 cases of venous ductus a wave flow reverse (4/9),3 cases of fetal nasal bone loss accompanied by tricuspid regurgitation and venous ductus a wave flow reverse (3/9).One case of nasal bone loss in 2 cases of trisomy 18,2 cases were tricuspid regurgitation and venous ductus a wave flow reverse.Two cases in 4 cases of 45X had venous ductus a wave flow reverse.There were 8 cases (0.16%) nasal bone absence in 4983 cases of normal karyotype fetus,48 cases (0.96%) of tricuspid regurgitation and 44 cases (0.88%) of venous ductus a wave flow reverse.Thirty-two cases in 40 cases (80%) of fetal congenital heart disease were tricuspid regurgitation,30 cases of venous ductus a wave flow reverse (75%).Eight cases of nasal bone absence normal karyotype fetus were found the nasal bone at 20 weeks gestation.Conclusion Combination screening of nuchal translucency with serum markers in the first trimester were high detection rate and low false positive rate; a wave reversion and fetal nasal bone absence accompanied by tricuspid regurgitation can improve the detection rate of abnormal karyotype; abnormalities ultrasound marker may be associated with fetal congenital heart disease at 11-13 +6 weeks of pregnancy.
3.Study on Compatible Stability of Voriconazole for Injection Combined with 2 Kinds of Injections
Xiaohong CUI ; Jinmin REN ; Meng LIU ; Zekun KANG ; Ying GONG
China Pharmacy 2017;28(17):2343-2346
OBJECTIVE:To investigate the compatible stability of Voriconazole for injection after mixed with Fructose injec-tion or Invert sugar injection. METHODS:Referring to package inserts,Voriconazole for injection 200 mg was dissolved with Wa-ter for injection to 20 mL,and then combined with Fructose injection 250 mL and Invert sugar injection 250 mL,respectively. At room temperature,the appearance of mixtures were observed 0,1,2,3,4,5 h after mixing,and pH value and the number of in-soluble particles were determined;the content of voriconazole was determined by HPLC. RESULTS:Under above condition,the appearance and pH value of mixtures had no significant change within 5 h;the number of particles ≥10 μm and ≥25 μm were all in line with the standard of Chinese Pharmacopoeia (2015 edition);the relative content of voriconazole was decreasing (95.28%-100%),but it changed within ±5%(RSD<2%,n=6). CONCLUSIONS:Voriconazole for injection could keep stable within 5 h after mixed with Fructose injection or Invert sugar injection.
4.Separation, purification and primary reverse cholesterol transport study of Cordyceps militaris polysaccharide.
Shou-Dong GUO ; Ying-Jie CUI ; Ren-Zhong WANG ; Ren-Yuan WANG ; Wen-Xue WU ; Teng MA
China Journal of Chinese Materia Medica 2014;39(17):3316-3320
The authors designed to separate, purify and determine the monosaccharide composition of the polysaccharide from Cordyceps militaris, and study its effect on reverse cholesterol transport in vivo by isotope tracing assay. Polysaccharides were separate and purify by ion exchange column Q-sepharose Fast Flow and size exclusion column Sephacryl S200HR; the molecular weight and monosaccharide composition of the polysaccharides were determined by high performance gel permeation chromatography and high performance liquid chromatography coming with pre-column derivation, respectively. Finally, three purified polysaccharides CMBW1, CMBW2 and CMYW1 were obtained, their total carbohydrate contents were 87%, 89%, 95%, respectively; their protein contents were 6.5%, 1.3%, 2.8%, respectively; their molecular weights were 772.1, 20.9, 13.2 kDa, respectively; CMBW1 was composed of mannose, glucosamine, rhamnose, glucuronic acid, glucose, galactose and arabinose with a molar ratio of 7.25: 0.17: 1.29: 0.23: 6.30: 11.08: 0.79; CMBW2 was composed of mannose, glucosamine, galactose and arabinose with a molar ratio of 2.40: 0.16: 2.92: 0.24; CMYW1 was composed of mannose, glucosamine, glucuronic acid and glucose with a molar ratio of 0.59: 0.57: 0.45: 25.61. Polysaccharide at 50 mg x kg(-1) could significantly improve the transport of 3H- cholesterol to blood and excretion from feces. All of the three purified polysaccharides CMBW1, CMBW2 and CMYW1 were heteropolysaccharide; and they could improve reverse cholesterol transport in vivo, the underlying mechanisms are being studied.
Animals
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Biological Transport
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drug effects
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Cholesterol
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metabolism
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Chromatography, High Pressure Liquid
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instrumentation
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methods
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Cordyceps
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chemistry
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Mice
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Monosaccharides
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analysis
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isolation & purification
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Polysaccharides
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chemistry
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isolation & purification
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pharmacology
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Tritium
5.Study on compatibility of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma based on pharmacokinetics of effective components salvianolic acid B and ferulic acid in rat plasma.
Cui-ying ZHANG ; Hong ZHANG ; Yu DONG ; Wei-guang REN ; Heng-wen CHEN
China Journal of Chinese Materia Medica 2015;40(8):1589-1593
A study was made on the pharmacokinetic regularity of effective components salvianolic acid B and ferulic acid in Salviae Miltiorrhizae Radix et Rhizoma (SMRR) and Chuanxiong Rhizoma(CR) in rats, so as to discuss the compatibility mechanism of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma. Rats were randomly divided into three groups and intravenously injected with 50 mg x kg(-1) salvianolic acid B for the single SMRR extracts group, 0.5 mg x kg(-1) ferulic acid for the single CR extracts group and 50 mg x kg(-1) salvianolic acid B + 0.5 mg x kg(-1) ferulic acid for the SMRR and CR combination group. The blood samples were collected at different time points and purified by liquid-liquid extraction with ethyl acetate. With chloramphenicol as internal standard (IS), UPLC was adopted to determine concentrations of salvianolic acid B and ferulic acid. The pharmacokinetic parameters of salvianolic acid B and ferulic acid were calculated with WinNonlin 6.2 software and analyzed by SPSS 19.0 statistical software. The UPLC analysis method was adopted to determine salvianolic acid B and ferulic acid in rat plasma, including linear equation, stability, repeatability, precision and recovery. The established sample processing and analysis methods were stable and reliable, with significant differences in major pharmacokinetic parameters, e.g., area under the curve (AUC), mean residence time (MRT) and terminal half-life (t(1/2)). According to the experimental results, the combined application of SMRR and CR can significantly impact the pharmacokinetic process of their effective components in rats and promote the wide distribution, shorten the action time and prolong the in vivo action time of salvianolic acid B and increase the blood drug concentration and accelerate the clearance of ferulic acid in vivo.
Animals
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Apiaceae
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chemistry
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Benzofurans
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blood
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pharmacokinetics
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Coumaric Acids
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blood
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pharmacology
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Drug Interactions
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Drugs, Chinese Herbal
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analysis
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pharmacokinetics
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Male
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Rats
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Rats, Sprague-Dawley
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Rhizome
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chemistry
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Salvia miltiorrhiza
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chemistry
6.Screening of Protein Interacting with HCMV UL130 Protein by Yeast Two Hybrid from Human Fetus Brain cDNA Library
Gaowei REN ; Xin CUI ; Ying QI ; Yanpin MA ; Qiang RUAN ; Zhengrong SUN
Journal of China Medical University 2010;(6):413-416
Objective To screen the human proteins interacting with human cytomegalovirus(HCMV)UL130 from human fetus brain cDNA library by GAL4 two-hybrid system 3 technique and analyze the corresponding coding sequences.Methods The "bait plasmid"(named as pGBKT7-UL130)was constructed.By using HCMV UL130 as the bait,a human fetus brain cDNA library was screened and the proteins interacting with UL130 protein were searched.The positive clones were sequenced and analyzed by bioinformatic methods.Results Nine clones interacting with HCMV UL130 were identified,two of them were synaptosome-associated protein(SNAP).Conclusion Some proteins interacting with HCMV UL130 in human fetus brain cDNA library were successfully screened.SNAP might play an important role in HCMV infection pathogenesis.
7.A Model Study: Simulation of Marker Gene Deletion and Target Gene Replacement Mediated by Cre/lox System in Cell Line
Wentao CUI ; Liming REN ; Jian HOU ; Ying ZHANG ; Yongfu CHEN ; Xiaorong AN
Progress in Biochemistry and Biophysics 2008;35(6):650-660
The incorporation of site-specific recombination systems can help to overcome bottlenecks in livestock transgenic technology. For evaluating the efficiency of Cre/lox mediated DNA recombination in embryos and somatic cells, a working model was established using rat mammary carcinoma cell line SHZ-88, aimed at creation of and use repeatedly of selected "friendly loci" in transgenic livestock. An integration vector pTE-lox2272-DsRed-loxP-GFP-loxP, which red fluorescence gene DsRed served as the first target gene and green fluorescence gene GFP as marker gene, was constructed for introduction of acceptor loci in genome. At the same time a replacement vector pT-lox2272-neo-loxP in which Neo coding sequence served as the second target gene was also constructed for replacing DsRed gene. Transgenic cell clones were produced by electroporating SHZ-88 cell with the integration vector. Cells from three transgenic clones selected randomly were further amplified and were then co-electroporated with the replacement vector as well as cre gene. Analysis of the expression patterns of DsRed and GFP indicated that among the 1 070 cell colonies the efficiency on marker GFP deletion was 91.1% and the efficiency on gene replacement was 29.3%. Molecular analysis by PCR and Southern blotting confirmed that the color patterns as expressed by cell colonies could represent the actual molecular events. This working model mediated by Cre/lox system should be useful for the improvement of the present animal transgenic technology.
8.Effect of low T_3 syndrome on outcome of acute myocardial infarction
qi, SHAO ; ying, REN ; jun-min, QUAN ; dong-mei, DU ; cui-chun, ZHAO ; meng, WEI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To investigate the prevalence of low T3 syndrome in patients with acute myocardial infarction(AMI) and explore the effect of low T3 syndrome on outcome of AMI.MethodsThree hundred and thirty-eight patients with AMI admitted to cardiac care unit(CCU) underwent examinations of thyroid function and cardial ultrasound,and were further categorized according to thyroid hormone profile.The records of noninvasive bi-level positive airway pressure(BiPAP)ventilation utilization,length of hospital stay,mortality during hospitalization were evaluated,and the related factors were analysed.ResultsOne hundred and thirty-nine of the 338 patients(41.12%) with AMI complicated with low T3 syndrome.Free triiodothyronine(FT3) was the independent influential factor for length of hospital stay.Low FT3 was significantly correlated with noninvasive BiPAP ventilation utilization and mortality during hospitalization.The average time of follow-up was(21.4?8.1) months.It was revealed by multivariate Cox regression analysis that FT3 was the chief predictor for cumulative death(risk ratio,4.25;95% confidential interval,2.30-7.87),followed by age and left ventricular ejection fraction.ConclusionThe recognition of AMI complicated with low T3 syndrome plays an important role in predicting the disease severity and outcome.
9.Screening of protein interacting with the transcript of UL128 gene showed two protein patterns by yeast two-hybrid from human fetus brain cDNA library
Gaowei REN ; Xin CUI ; Yanping MA ; Ying QI ; Qiang RUAN ; Zhengrong SUN
Chinese Journal of Microbiology and Immunology 2010;30(8):737-742
Objective Using yeast two-hybrid system to screen the proteins which can interact with the human cytomegalovirus (HCMV) UL128 which have two difference transcription structure from human fetus brain cDNA library, and compare the difference with structure and function of interacting proteins. Methods Two fragments of UL128 were amplified by 3'RACE and 5'RACE technology, the length are 519 bp and 642 bp, respectively. The "bait plasmid" (named as pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp) was constructed successfully. Using pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp as a bait, a human fetus brain cDNA was screened and the proteins interacting with UL128-519 bp and UL128-642 bp encoded protein were searched, and the positive clones were sequenced and analyzed by bioinformatic methods. Results EFEMP2 interacting with HCMV UL128-519 bp were identified, THY-1 interacting with HCMV UL128-642 bp were identified. Conclusion EFEMP2 and THY-1 proteins interacting with HCMV UL128-519 bp and UL128-642 bp in human fetus brain cDNA library were successfully screened, but same proteins weren't found from the proteins interacting with UL128-519 bp and UL128-642 bp protein, UL128-519 bp and UL128-642 bp protein may be play an different effect in the process of infect by HCMV.
10.Association between human leukocyte antigen DQB1 gene polymorphisms and bronchial asthma among Mongolia and Han nationalities
Liying CUI ; Ying NIAN ; Jie TAN ; Hong SUO ; Xiuyun Lü ; Tianji ZHU ; Hui REN
Chinese Journal of General Practitioners 2010;09(9):619-623
Objective To investigate association between human leukocyte antigen DQB1 (HLADQB1 ) gene polymorphisms and bronchial asthma among Mongolian and Han nationalities. Methods Sequence-specific primer polymerase chain reaction (PCR-SSP) was used to detect frequencies of HLA DQB1 genotypes and alleles in 50 cases of Han and 68 Mongolian asthmatic patients, and 50 Han and 54 Mongolian healthy controls, respectively. Difference in gene frequencies between the two nationalities was estimated by odds ratio (OR) and chi-square test. Results Frequency of the HLA-DQB1 0602 allele was significantly higher in Han patients with bronchial asthma than that in healthy Han nationality (OR = 6.163,P <0.01 ). Frequency of the HLA-DQB1 0603/0608 allele decreased in Mongolian asthmatic patients, as compared to that in healthy Mongolians ( OR = 0.199, P < 0.05 ). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian asthmatic patients as compared to that in healthy Mongolians ( OR =2.074,P <0.05). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian than that in Han asthmatic patients ( OR = 2.482 ,P =0.05). Frequency of the HLA- DQB1 0602 allele was significantly higher in healthy Mongolians than that in healthy Han nationality ( OR = 3.341, P < 0.05 ), in contrast, frequency of the HLA-DQB1 0402 allele was significantly lower in healthy Mongolians than that in healthy Han nationality ( OR = 0.209, P < 0.05 ). Conclusions The HLA-DQB1 0603/0608 allele is possibly a protective gene and the HLA-DQB1 0301/4 allele a susceptible gene for bronchial asthma in Mongolians, and the HLA-DQB1 0602 allele is possibly a susceptible gene for bronchial asthma in Han nationlity.