简介：AbstractThere is increasing evidence that cell-free DNA (cfDNA) in spent culture media (SCM) can be amplified for genetic testing. Therefore, this paper reviews the characteristics of cfDNA, including its fragment size, amount, origin, as well as some factors affecting the success rate of its amplification, together to provide researchers with a more comprehensive perspective on embryonic cfDNA. The origin of cfDNA in SCM is complicated and poses challenges to the interpretation of genetic test results. Advanced molecular techniques should distinguish between embryonic and contaminated DNA to maximize the success rate of amplification and analysis. Recent data showed that the type of culture medium, assisted hatching or not, the type of amplification kit, and fresh or thawed embryos were not related to the success rate of amplification, but the length of culture time might affect the success rate. The longer culture time, the more cfDNA is available in the SCM. Then we focused on the concordance between trophectoderm (TE), inner cell mass, whole embryo, and embryonic cfDNA. Despite successful amplification, the concordance between TE and embryonic cfDNA was low. In summary, non-invasive genetic testing using SCM could represent a major advance in future single embryo selection, however, contamination and timing for media collection are key factors affecting the results, and current non-invasive cfDNA testing should not be directly applied to clinical practice. Further research is needed to improve the methods used for testing techniques and genetic analysis to achieve greater accuracy and trace its origins before it can be used in the clinics.

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简介：AbstractObjective:We aim to assess the clinical performance of cell-free fetal DNA (cffDNA) testing for detecting common fetal aneuploidies as well as subchromosomal deletions/duplications and explore the pregnancy decisions in screen-positive cases.Methods:A cohort of 19,531 pregnant women was offered cffDNA testing for detection of trisomies 21, 18, and 13 (T21, T18, and T13); sex chromosome aneuploidies (SCAs); and subchromosomal deletions/duplications. Screen-positive cases were confirmed by karyotyping and single-nucleotide polymorphism array analysis.Results:A total of 47 cases failed the test. The overall screen-positive rate of chromosomal abnormalities was 1.07% (208/19,484), including 57 cases with T21, 18 cases with T18, 7 cases with T13, 106 cases with SCAs, and 20 cases of subchromosomal deletions/duplications. Positive predictive values were 91.30% (42/46), 38.46% (5/13), 33.33% (2/6), 41.33% (31/75), and 27.78% (5/18), respectively. There was no significant difference in the screening of fetal chromosomal aneuploidies in the high-risk group compared with the low-risk group (P > 0.05). All of the pregnant women who had confirmed fetal T21, T18, or T13 terminated their pregnancies, except for a case of T13 mosaic, whereas 45.16% (14/31) of women with fetal SCAs continued their pregnancies. Furthermore, 17 pregnant women with positive screens for T21, T18, or T13 without a subsequent diagnosis chose to terminate their pregnancy, whereas 29 of 31 women with SCAs chose to continue their pregnancies.Conclusions:CffDNA testing exhibited good screening accuracy for T21, T18, and T13 and also contributed to detecting fetal SCAs and subchromosomal deletions/duplications. Pregnant women with fetal 47, XXX or 47, XYY were more willing to terminate their pregnancy than those with fetal 45, X or 47, XXY.

简介：AbstractPreimplantation genetic testing (PGT) is a widely adopted screening method that can be performed to identify and select embryos with normal ploidy; however, PGT relies on embryo biopsy, that is, polar body, embryo cells, or trophectoderm biopsy, to obtain embryonic DNA, increase its technical limitations. Studies have indicated that biopsy may have an influence on the quality and development of embryos, and increase the chance of abnormal epigenetic modifications. Therefore, non-invasive PGT (niPGT) detection of cell-free DNA (cfDNA) has gradually become a hot research topic in the field of assisted reproduction. Studies showed cfDNA could be detected in blastocyst fluid and spent culture medium (SCM) in vitro cultured embryos. The cfDNA collection requires less skill and makes lower risk to embryos. Some studies have been conducted to evaluate the feasibility of SCM-based niPGT approaches. When comparing the ploidy consistency of cfDNA in SCM, its consistency to the conventional PGT for aneuploidies results fluctuated widely, it is critical to recognize the factors influencing accuracy. These contradictory results may be related to factors such as the difference in SCM sampling methods and sampling time, and the definition of consistency. In this review, we aimed to comprehensively summarize how researchers use embryonic cfDNA to conduct niPGT detection. It also systematically reviews the factors affecting the accuracy of the test and its underlying issues, as well as prospective applications. We hope to provide a basis for future niPGT research and a useful reference for the standardized operation of niPGT that can be widely applied in clinical practice.

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简介：AbstractBackground:Deregulation of miRNA-21 expression has been reported to be associated with vascular smooth muscle behavior and cytoskeletal stability. This study is aimed to investigate the density of serum miRNA-21 in patients with different phases of intracranial aneurysms (IAs) and explore its warning function for IA rupture.Methods:A total of 16 in 200 IA patients were selected and categorized into 4 groups based on the phase of IA. Microarray study was carried out using serum miRNA and differentially expressed miRNAs were identified. Another 24 samples from a cohort of 360 patients were added and real-time polymerase chain reaction (RT-PCR) was performed on expanded sample size (n = 40) for miRNA-21 validation. Potential gene targets of miRNA-21 were screened out from Gene Ontology (GO) database and literatures.Results:Microarray study identified 77 miRNAs with significantly different expression levels between experimental groups and the control group. RT-PCR assays validated significant downregulation of miRNA-21 in experimental groups, among which miRNA-21 expression level of daughter aneurysm group decreased the most. Bioinformatic analyses revealed that several target genes related with miRNA-21 may be involved in IA formation and rupture.Conclusions:This study suggested that miRNA-21 had a protective effect for intracranial vascular wall against remodeling and warning function for intracranial aneurysm rupture. Significant suppression of serum miRNA-21 in IA patients may provide diagnostic clues for aneurysm rupture and guide clinical intervention.

简介：AbstractOver the past 50 years, the scope and extent of prenatal diagnosis and screening for genetic disorders have improved geometrically. There has been a pendulum like swing from testing to screening back and forth as new technologies emerge. The concurrent developments of cell free fetal DNA analysis of maternal blood has dramatically changed patient’s choices towards screening. However, with the use of array comparative genomic hybridization of fetal DNA that requires diagnostic procedures (Chorionic villus sampling and amniocentesis), much more extensive diagnosis can be obtained. Until noninvasive methods can replicate what can be done with diagnostic procedures there still will be a "price to be paid" for opting for the non-invasive methods.

简介：AbstractObjective:To evaluate the screening performance of noninvasive prenatal testing (NIPT) based on high-throughput massively parallel sequencing technology for the fetal XXY aneuploidies among pregnancies in Beijing of China.Methods:The study enrolled 26 913 consecutive pregnancies, 20-50 years old, who attended the Peking Union Medical College Hospital, Beijing, China, for prenatal screening from January 1, 2016 to December 31, 2019. Cell-free DNA was extracted from maternal peripheral blood to have a high-throughput massively parallel sequencing procedure. Cases with high-risk of fetal XXY were suggested to take invasive prenatal diagnosis (IPD) for confirmation. Maternal DNA sequencing was performed, if necessary, to find other potential factors that may lead to high-risk results of XXY by NIPT.Results:Among a cohort of 26 913 pregnant women, 34 were high-risk for fetal XXY, among which 30 accepted IPD while 4 declined. In those who accepted IPD, 19 cases were confirmed fetal XXY by chromosome karyotyping analysis while 11 were verified as false positive. Among the 19 confirmed fetal XXY cases, 14 elected pregnancy termination. For all the 34 high-risk cases, two were verified maternal sex chromosome aneuploidy. The calculated detection rate, positive predictive value, and false-positive rate of NIPT for fetal XXY in this cohort was 100.00% (19/19), 63.33% (19/30), and 0.04% (11/26 890), respectively. And the percentage of pregnancy termination was 73.68% (14/19).Conclusion:NIPT could be used as a potential method for fetal XXY screening, although the accuracy needs to be improved. As NIPT is not diagnostic, IPD is strongly recommended for those with high-risk results. For cases with discordance between NIPT and fetal karyotyping, maternal DNA sequencing would help to identify the cause of false-positive/false-negative results.

简介：【摘要】目的：探究无创DNA产前检测（non-invasive prenatal testing，NIPT)在高龄孕妇胎儿非整倍体染色体疾病产前诊断中的临床应用价值，为高龄孕妇进行无创DNA临床应用提供依据。方法：收集2018年1月至2022年10月在我院自愿进行无创DNA产前检测的单胎高龄孕妇1183例，按年龄将孕妇分为35-39岁组(n=1100)与年龄≥40岁组(n=83)。采用NIPT高通量测序检测孕妇外周血中胎儿游离DNA。检测结果提示染色体高风险者行羊膜腔穿刺抽取羊水进行胎儿染色体核型分析。对检测结果提示低风险者通过电话随访进行验证。观察其胎儿染色体非整倍体疾病诊断。结果：1183例受检者NIPT检测结果提示，胎儿非整倍体染色体异常者12例，其中，21-三体5例，18-三体2例，13-三体1例，性染色体异常4例（2例47，XXX/XXY；1例47，XYY；1例45，XO）。NIPT检测在高龄孕妇中阳性率为1.01%(12/1183)。12例高风险孕妇其中自然流产1例，其余11例均做羊水穿刺，5例21-三体高风险均经羊水穿刺确诊，21-三体在高龄孕妇中的阳性预测值达到100%(5/5)，18-三体在高龄孕妇中的阳性预测值为50%(1/2)，1例13-三体经羊水穿刺排除，性染色体阳性预测值为50%(2/4)。35～39岁年龄组染色体异常高风险发生率为0.73%(8/1100)，年龄≥40岁组染色体异常高风险发生率为4.82%（4/83），两组比较，差异有统计学意义(P＜0.05﹚。结论：NIPT检测对高龄孕妇胎儿 21-三体、18-三体及性染色体符合率较高，可明显减少高龄孕妇介入性产前诊断，但对 13- 三体和性染色体数目减少的符合率偏低，NIPT检测异常的孕妇仍需行羊水穿刺确诊，避免不必要的流产。

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简介：Objective:ToexploretherelationshipbetweenquantitativeTreponemapallidumDNA(TP-DNA)PCRtestingandtheToludineRedUnheatedSerumTest(TRUST)inpatientswithsyphilisbeforeandaftertreatment,andevaluatetheclinicalvalueofquantitativeTP-DNAtestinginthediagnosisandtreatmentevaluationofsyphilis.Methods:29patientswithprimary(12cases)orsecondary(17cases)syphilis,whometthecriteriasetforthisstudywererecruitedassubjects.Allpatientsweretreatedwith2.4millionunitsbenzathinepenicillinIMweeklyfor3weeks.QuantitativetestsofTP-DNAinthepatients'plasmawereperformedusingFQ-PCRbeforeandafterthetreatment.SerologictestsincludingTRUSTandTPPAwerealsoperformed.Results:Beforethetreatment,9outof12primarysyphilispatients(75%)andallsecondarysyphilispatients(17/17)testedpositiveforTreponemapallidum(TP)byTP-DNAtesting.TheaveragequantitativetestvaluesofTP-DNAinprimaryandsecondarysyphilispatientswere(3.38±2.34)×10^4and(5.73±1.33)×10^6copies/ml,respectively.Afterthreemonthsoftreatment,1ofthe9primaryand5outof17secondarysyphilispatientswerepositiveuponTP-DNAtesting,respectively.TheaveragequantitiesofTP-DNAwere2.01×10^2copies/mlinprimaryand5.87×10^2copies/mlinsecondarysyphilispatientswithpositiveTRUSTandTP-DNAtests,and3.09×10^2copies/mlforthosewithnegativeTRUST,respectively.Afterninemonthsoftreatment,alltheprimaryandsecondarysyphilispatientswerenegativeuponTP-DNAtesting,whileallprimaryand14of17(82.35%)secondarysyphilispatientsshowednegativeTRUSTresults.Conclusion:ThattheresultsofTP-DNAtestsarenotconsistentwiththoseofTRUSTbeforeandaftertreatmentindicatesthatquantitativeTP-DNAtestingmayhavevaluableclinicalsignificanceintheearlydiagnosisandevaluationoftreatmentregimensforsyphilis.

简介：TheDNAcontentandmorphometricfeaturesofhepatocellularcarcinoma(HCC)andlivercelldysplasia(LCD),includingnucleararea,nuclearperimeter,nuclearmaximumdiameterandnuclearcirclediameter,werequantitativelydeterminedbymeansofimageanalysistechnology.Theresultsshowedthatincomparisonwithnormalhepatocytes,LCDhadamarkedlyincreasedDNAcontentandnuclearmorphometricparameters,butthevalueswerelowerthanthoseforHCC.LCDshowedaslightincreaseinnuclearatypiarepresentedbythenuclearirregularindex,whichwasalsolessthanHCC.ThefindingsindicatethatLCDmaybeaprecaneerouslesionofHCC,tothecellsinanabnormalproliferativestate.

简介：IncubationofdinoflagellateCrythecodiniumcohniichromosomesincytoplasmicextractsofunfertilizedXenopuslaeviseggsresultedinchromosomesdecondensationandrecondensation,nuclearenvelopeassembly,andnuclearreconstitution.DinoflagellateCrythecodiniumcohniiisakindofprimitiveeukaryotewhichpossessesnumerouspermanentlycondensedchromosomesanddiscontinuousdouble-layerednuclearmembranethroughoutthecellcycle.Theassemblednuclei,beingsurroundedbyacontinuousdoublemembranecontainingnuclearporesandtheuniformlydispersedchromatinfibersaremorphologicallydistinguishablefromthatofDinoflagellateCrythecodiniumcohnii.However,incubationofdinoflagellateCyrthecodiniumcohniichromosomesintheextractsfromdinoflagellateCrythecodiniumcohniicellsdoesnotinducenuclearreconstitution.

简介：ObjectivesTodetectionofchlamydiapneumoniae(Cpn)DNAinthecirculatingmononuclearcellfractionsofcoronaryheartdiseaseandtoinvestigatetheassociationbetweeninfectionwithchlamydiapneumoniaeandcoronaryheartdisease(CHD)andprospectivelywhetherblood-basednestedpolymerasechainreaction(nPCR)isusefulinidentifyingCpninfection.MethodsTheperipheralbloodmononuclearcell(PBMC)CpnDNAwasexaminedusingnPCRtechniqueandconfirmedbyelectrophoresisin150patientswithCHD.Select55patientswithclinicalsuspectedCHDbutangiographyresultarenormalascontrolgroup(CG).Thenweconductedaprospective,randomized,double-blind,placebo-controlledstudyof6monthsofazithromycinandplacebotreatmentinCHDgroup.PatientswithCpnDNApositivewerethenrandomizedtoreceiveazithromycinorplacebo.Aftertreatmentbloodsamplewerecollectedforrepeatedmeasurement.ResultsChlamydiapneumoniaeDNAwasdetectedin49(32.7%)of150personswithCHDandin1(1.8%)of55personswithcontrolgroup,oddsratio26.2,95%confidenceinterva13.52-194.98.ThepositivityratesofnPCRinCHDgroupswerehigherthanthoseincontrolgroup.16cases(29.1%)inlatentcoronaryheartdiseases(LCHD)group,19cases(39.6%)inunstableangina(UAP)group,and14cases(29.9%)inacutemyocardialinfarction(AMI)groupwereCpnpositivebynPCR.TherewerenosignificantdifferenceamonginAMIUAPandLCHDgroup.ThereweresignificiantdifferenceinCpnDNAnegativeratesaftertheazithromycinandtheplacebotreatment.ConclusionsChlamydiapneumoniaeispresentinPBMCofasignificantproportionofpersonswithCHD.Thepotentialroleofchlamydiapneumoniaeincoronaryatherosclerosismaythereforebemorerelatedtoaccelerationofdiseaseorsystemiceffectsbypersistentinfectionthantosuddeninitiationofprogressivecoronaryarterydiseasebyacuteinfection.ThedetectionofCpnDNAinPBMCwithnPCRmaybeofgreatvalueforidentifyingCpncarriersandfo

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简介：<正>DearSir,IamDongHyunJi,fromtheDepartmentofOphthalmologyofSt.Vincent’sHospital,Suwon,Korea.Iwritetopresentaveryseverelyrecurrentbasalcellcarcinoma(BCC)inlowerlidinvadingleftorbitandwholehemiface,

简介：Inthispaper,wetooktheleadinstudyingonspecificityofthemicrosatelliteDNAlociandapplicabilityofmicrosatelliteDNAprimersinprotozoa.Inordertostudycharactersofmicrosatellitesinfree-livingprotozoa,eightmicrosatellitelociprimersdevelopedfromTrypanosomacruzi(MCLE01,SCLE10,MCLE08,SCLE11,MCLF10,MCLG10,MCL03,MCL05)wereemployedtoamplifymicrosatelliteinfourfree-livingprotozoa,includingBododesignis,EuglenagracilisFACHB848,ParameciumbruziseandTetrahymenathermophilaBF1.IntheamplificationsystemsofP.bruzise,fourloci(SCLE10,SCLE11,MCLF10,MCL03)wereamplifiedsuccessfully,andfouramplificationfragmentswereinpropersize.IngenomeofE.gracilisFACHB848,fiveofeightprimersbroughtfiveclearamplificationbands.InB.designis,three(No.4,5and7)ofeightlociproducedclearandsharpproductswithoutstutterbands,whereasnobandsappearedinT.thermophilaBF1.Further,eight300-500bpamplificationfragmentswereclonedandsequenced.Nevertheless,allsequencedproductsdidnotcontaincorrespondingmicrosatellitesequence,althoughBodoisinthesameorderandhasthenearestphylogeneticrelationwithTrypanosomaamongthesefourspecies.Thus,themicrosatelliteDNAprimerscannotbeappliedamongorderormorefartaxa,andthespecificityofmicrosatelliteDNAisveryhighinprotozoa.TheresultsofthisstudywillcontributetoourunderstandingofmicrosatelliteDNAinprotozoa.

简介：AbstractHarlequin ichthyosis is a severe autosomal recessive skin disorder. Most deaths occur within the first few days after birth, and the survivors still have severe chronic skin disease throughout their lives. Almost all cases were associated with a pathogenic variant of adenosine triphosphate binding cassette transporter, subfamily A, member 12 (ABCA12) gene. We described a case of HI diagnosed by ultrasound examination during the second-trimester and genetic diagnosis reveal two novel heterozygous ABCA12 mutations c.2563-2570delinsGGCAATT, p.(Leu855Glyfs*13), and c.6116delT, p.(Met2039Argfs*8) by the next-generation DNA sequencing, which further enriched our understanding of the pathogenic variation of ABCA12 gene.

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