Objective:To assess whether personalized embryo transfer guided by endometrial receptivity array(ERA)improves implantation and pregnancy rates in women with implantation failure.Methods:This retrospective case-control...Objective:To assess whether personalized embryo transfer guided by endometrial receptivity array(ERA)improves implantation and pregnancy rates in women with implantation failure.Methods:This retrospective case-control study was conducted on women with previous implantation failure.The women were divided into two groups,i.e,women who underwent ERA and those who underwent embryo transfer without ERA testing.ERA was performed using Igenomix.ERA results were interpreted as receptive or non-receptive.Women underwent frozen embryo transfer on the 6th day of progesterone(P+5).The primary outcomes were implantation rate,clinical pregnancy rate,abortion rate,and negative pregnancy rate.Results:This study included 229 women with previous implantation failure,with 154 in the ERA group and 75 in the no ERA group.The mean age of the women of the ERA group was(32.2±4.1)years,and that of the no ERA group was(31.5±4.8)years.Women in the ERA group had a higher implantation rate(60.4%)and clinical pregnancy rate(57.1%)compared to those in the no ERA group(48.0%and 46.7%,respectively).In addition,implantation rate of the nonreceptive ERA group was higher than the no ERA group(65%vs.48%),and clinical pregnancy rate was also higher in the non-receptive ERA group than the no ERA group(65%vs.47%).The abortion rate of the no ERA group was 9%and that of the non-receptive ERA group was 10%.52%no ERA group women and 35%non-receptive ERA group women had negative pregnancy results.Conclusions:Women who have undergone personalised embryo transfer guided by ERA have a higher clinical pregnancy rate than women who have not after previous implantation failure.展开更多
Background:Four-dimensional(4D)ultrasound is increasingly being used for prenatal diagnosis of congenital heart disease(CHD).We aimed to perform a systematic review and meta-analysis to evaluate its diagnostic accurac...Background:Four-dimensional(4D)ultrasound is increasingly being used for prenatal diagnosis of congenital heart disease(CHD).We aimed to perform a systematic review and meta-analysis to evaluate its diagnostic accuracy for fetal CHD.Methods:This systematic review was conducted in accordance with the PRISMA-DTA guidelines.We systematically searched eight databases for studies published up to July 22,2025.Data were extracted to calculate diagnostic accuracy metrics,study quality was assessed using QUADAS-2,and a bivariate random-effects model was used for the meta-analysis.Results:A total of 49 studies were included,comprising 45 retrospective and 4 prospective studies,which were mainly(91.8%)conducted in China.These studies involved 23,397 fetuses,among which 2115 were diagnosed with congenital heart disease.The pooled sensitivity of 4D ultrasound for diagnosing fetal congenital heart disease was 0.91(95%CI:0.89-0.93),the pooled specificity was 0.98(95%CI:0.97-0.99),and the area under the summary receiver operating characteristic(SROC)curve(AUC)was 0.98(95%CI:0.96-0.99).Although 4D ultrasound technology can be implemented as early as 11 weeks of gestation,its diagnostic sensitivity and specificity reached a superior level and stabilized at an average of 20 weeks of gestation(range 14-28 weeks).Meta-regression indicated that sample size and prior suspicion of fetal CHD were significant contributors to heterogeneity(p<0.05).Conclusion:Four-dimensional ultrasound has high diagnostic efficacy for fetal CHD and is suitable for prenatal screening of fetal CHD,and the diagnostic effect is optimal and stable at an average gestational age of 20 weeks(range 14-28 weeks).展开更多
Preterm birth(PTB),defined as delivery before 37 weeks of gestation,is the most common adverse pregnancy outcome[1].PTB is a global health concern,with an estimated 13.4 million cases in 2020[1],accounting for more th...Preterm birth(PTB),defined as delivery before 37 weeks of gestation,is the most common adverse pregnancy outcome[1].PTB is a global health concern,with an estimated 13.4 million cases in 2020[1],accounting for more than one in 10 births worldwide.Compared to full-term births,PTBs are associated with a higher risk of short-and long-term complications,including bronchopulmonary dysplasia,necrotizing enterocolitis,visual impairment,and cerebral injuries[2].Despite substantial research efforts to prevent PTB,the global PTB rate has shown little improvement over the past decade[1].Therefore,identifying additional risk factors remains a critical goal in preventing PTB.展开更多
Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(R...Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(Reiter and Leroux,2017;Wallmeier et al.,2020).To date,over 180 ciliopathy-associated genes have been identified(Reiter and Leroux,2017),yet the underlying mechanisms remain poorly understood.展开更多
Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the...Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the human endometrial epithelial cells(HEECs)was explored.Methods HEECs were cultured in copper-containing culture medium to simulate changes in the endometrium after copper intrauterine device(Cu-IUD)implantation.Reverse transcription quantitative PCR(RT-qPCR)was used to detect the differential expression of miR-144-3p in HEECs after Cu^(2+)treatment.MiRNAs,siRNAs and related inhibitors were used to treat HEECs.The expression levels of related downstream genes were then analyzed by RT-qPCR,Western blotting and immunofluorescence to explore the specific mechanism involved.Results MiR-144-3p was significantly upregulated in the Cu^(2+)-treated HEECs.The expression of P-NF-κB,MMP9,TGF-β3 and P-SMAD3 was significantly decreased in HEECs treated with 10μg/mL Cu^(2+).MiR-144-3p regulated the expression of metallothionein 1A(MT1A)and thrombospondin-1(THBS-1)in Cu^(2+)-treated HEECs.The expression of P-NF-κB can be regulated by MT1A,and an inhibitor of P-NF-κB can significantly reduce the expression of MMP9 in Cu^(2+)-treated HEECs.The expression of TGF-β3 can be regulated by THBS-1,and a TGF-β3 inhibitor can significantly reduce the expression of SMAD3 in Cu^(2+)-treated HEECs.The proliferative capacity of HEECs treated with MMP9 or SMAD3 inhibitors was significantly reduced.Conclusions The increased Cu^(2+)concentration led to the upregulation of miR-144-3p,further reducing the expression levels of its target genes(MT1A and THBS-1),which in turn downregulated the expression of NF-κB,MMP9,TGF-β3 and SMAD3,ultimately leading to increased endometrial cell damage and decreased cell proliferation.展开更多
DNA methylation is an important promising biomarker for cancer diagnosis and monitoring.Therefore,the assessment of DNA methylation levels is helpful for the prognosis and diagnosis of cancer.However,it is still a hug...DNA methylation is an important promising biomarker for cancer diagnosis and monitoring.Therefore,the assessment of DNA methylation levels is helpful for the prognosis and diagnosis of cancer.However,it is still a huge challenge to sensitively and accurately quantify the levels of DNA methylation in clinical sample.In this work,we proposed a protospacer adjacent motif(PAM)-free mediated CRISPR-Cas12a ultra-sensitive and quantitative DNA methylation detection method.Through recognizing the ds DNA with toehold region,CRISPR-Cas12a not only got rid of the limitation of PAM,but also improved its distinction ability for single Cp G site methylation,nearly 5-fold that of conventional PAM-containing ds DNA.We further introduced assist-strand and design an artificial mismatch to greatly improve the ability to distinguish single Cp G methylation site.Our results showed that the discrimination factor was >200.Then,we constructed toe-ds DNA by using “heating and freezing”,which made our method universally applicable and feasible.In addition,we greatly simplified the difficulty of primer design.Our method detected four highly methylated genes acyl carrier protein(ACP),CLV3/ESR-related(CLE),Disabled(DAB) and Homeobox(HOX) with a detection limit of 0.01 % and excellent linearity in DNA methylation standards.Then,we verified the clinical utility of this method in 29 hepatocellular carcinomas,11 ovarian cancers and4 health people.In conclusion,we have successfully constructed a PAM-free CRISPR-Cas12a DNA methylation quantification method,which achieves high congruence in sensitivity,specificity and universality,fully demonstrating its significant clinical application value.展开更多
Male reproductive infections are known to shape the immunological homeostasis of the testes,leading to male infertility.However,the specific pathogenesis of these changes remains poorly understood.Exosomes released in...Male reproductive infections are known to shape the immunological homeostasis of the testes,leading to male infertility.However,the specific pathogenesis of these changes remains poorly understood.Exosomes released in the inflammatory microenvironment are important in communication between the local microenvironment and recipient cells.Here,we aim to identify the immunomodulatory properties of inflammatory testes-derived exosomes(IT-exos)and explore their underlying mechanisms in orchitis.IT-exos were isolated using a uropathogenic Escherichia coli(UPEC)-induced orchitis model and confirmed that IT-exos promoted proinflammatory M1 activation with increasing expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in vitro.We further used small RNA sequencing to identify the differential miRNA profiles in exosomes and primary testicular macrophages(TMs)from normal and UPEC-infected testes,respectively,and identified that miR-155-5p was highly enriched in IT-exos and TMs from inflammatory testes.Further study of bone marrow derived macrophages(BMDMs)transfected with miR-155-5p mimic showed that macrophages polarized to proinflammatory phenotype.In addition,the mice that were administrated IT-exos showed remarkable activation of TM1-like macrophages;however,IT-exos with silencing miR-155-5p showed a decrease in proinflammatory responses.Overall,we demonstrate that miR-155-5p delivered by IT-exos plays an important role in the activation of TM1 in UPEC-induced orchitis.Our study provides a new perspective on the immunological mechanisms underlying inflammation-related male infertility.展开更多
Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ...Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ cell development.Heterogeneous nuclear ribonucleoproteins(hnRNPs)are a large family of RBPs implicated in many steps of RNA processing;however,their roles in spermatogenesis are largely unknown.Here,we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis.Furthermore,we found that most of the detected hnRNP proteins(hnRNPD,hnRNPK,hnRNPQ,hnRNPU,and hnRNPUL1)display the highest signals in the nuclei of pachytene spermatocytes,round spermatids,and Sertoli cells,whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids.The expression of these hnRNP proteins showed both similarities and specificity,suggesting their diverse roles in spermatogenesis.展开更多
Objective:To investigate the association between age and semen parameters among male partners of subfertile couples.Methods:This retrospective study analyzed the semen of 1523 infertile men aged 26 to 50 years.Data we...Objective:To investigate the association between age and semen parameters among male partners of subfertile couples.Methods:This retrospective study analyzed the semen of 1523 infertile men aged 26 to 50 years.Data were extracted from GarbhaGudi IVF Centre database from January 2019 to September 2020.The basic semen parameters were interpreted according to the WHO manual 2021,6th edition.Semen parameters in different age groups were compared.Results:Total and progressive motile sperms were significantly higher in the age group of 26-30 years compared to other age groups(P<0.05).Normal sperm count was significantly higher in the age group of 26-30 years compared to the age groups of 41-45 years and>46 years(P=0.001).However,sperm head defects,neck and midpiece defects,tail defects,and cytoplasmic droplets showed statistically insignificant difference in all the age groups(P>0.05).Semen viscosity showed no statistical difference in all the age groups compared to the reference age group of 26 to 30 years.Conclusions:Higher age can lead to a significant decrease in normal sperms and motility in subfertile men.Hence,male partner age should be considered as one of the major determining factors for reproductive outcomes.展开更多
DNA-functionalized gold nanoparticles are one of the most versatile bionanomaterials for biomedical and clinical diagnosis. Herein, we discovered that the performance of DNAzyme cleaving the substrate is highly relate...DNA-functionalized gold nanoparticles are one of the most versatile bionanomaterials for biomedical and clinical diagnosis. Herein, we discovered that the performance of DNAzyme cleaving the substrate is highly related to its length. This intriguing phenomenon only appears at the interfaces of DNAfunctionalized gold nanoparticles. We systematically investigated the causes of this phenomenon. We conjectured that the DNAzyme with extended nucleotides that do not match its substrate strand is vulnerable to non-specific adsorption, electrostatic repulsion, and steric hindrance. Based on our improved understanding of this phenomenon, we have successfully developed a highly sensitive and specific amplifiable biosensor to detect human apurinic/apyrimidinic endonuclease 1.展开更多
We presented a low-abundance mutation detection method with lambda exonuclease and DNA threeway junction structure.The assistant strand in the DNA three-way junction structure could regulate the reaction system from t...We presented a low-abundance mutation detection method with lambda exonuclease and DNA threeway junction structure.The assistant strand in the DNA three-way junction structure could regulate the reaction system from the kinetics and thermodyna mics aspects.The optimization of the assista nt strand helps to improve the selectivity of the mutant-type DNA to the wild-type DNA about 35 times.Moreover,the cost of the optimization process could be saved by about 90%.The method was applied to the detection of a human ovarian cancer-related gene mutation BRCA1(rs1799949,c.2082 C>T).The limit of detection to the mutation abundance in the DNA three-way junction structure system(0.2%) was one order lower compared with that in the double-stranded DNA structure system(2%).The mutation abundance in different standard samples was quantitively measured,and the results were consistent with the initial abundance in the standard samples.展开更多
Infections and inflammatory reactions in the male genital tract are the leading causes of male infertility with a prevalence of 6%-10%,primarily affecting testicular and epididymal function and ultimately compromising...Infections and inflammatory reactions in the male genital tract are the leading causes of male infertility with a prevalence of 6%-10%,primarily affecting testicular and epididymal function and ultimately compromising sperm quality.However,most infertile patients with genital infection/inflammation are asymptomatic and easily overlooked.Traditional indicators,including white blood cells,elastase,and other components in semen,can reflect inflammation of the genital tract,but there is still a lack of a uniform standard method of detection.Therefore,it is necessary to explore reliable markers in semen that reflect the inflammatory status of the genital tract.Using the experimental autoimmune orchitis(EAO)model to simulate noninfectious chronic orchitis,we successfully collected ejaculated seminal fluid from EAO rats using optimized electrical stimulation devices.Proteomic analysis was performed using isobaric tags for relative and absolute quantification(iTRAQ).Compared to the control group,55 upregulated and 105 downregulated proteins were identified in seminal plasma samples from the EAO group.In a preliminary screening,the inflammation-related protein S100A8/A9 was upregulated.We further verified that S100A8/A9 was increased in seminal plasma and highly expressed in testicular macrophages of the EAO model.In patients with oligoasthenospermia and genital tract infections,we also found that S100A8/A9 levels were remarkably increased in seminal plasma and testicular macrophages.S100A8/A9 in semen may be a potential biomarker for chronic genital inflammation.Our study provides a new potential biomarker for early diagnosis and further understanding of male infertility caused by genital inflammation.展开更多
Sperm damage caused by reactive oxygen species(ROS) is one of the main causes of male infertility.Therefore, the level of ROS in sperm is an important indicator for the diagnosis and prognosis of male infertility. Her...Sperm damage caused by reactive oxygen species(ROS) is one of the main causes of male infertility.Therefore, the level of ROS in sperm is an important indicator for the diagnosis and prognosis of male infertility. Herein, we constructed a single sperm ROS detection method(SSRDM) with an optical microprobe fabricated via focused ion beam process. The micro-probe is used to separately excite fluorescence in the sperm and the area around the sperm after ROS staining, and the difference in fluorescence values can reflect the level of ROS in the sperm. We collected 102 semen samples and 72 of them were divided into asthenozoospermia and non-asthenozoospermia groups. SSRDM and flow cytometry were used to detect the ROS levels of the two groups. The results of SSRDM showed that the ROS levels of asthenozoospermia group were higher than that of non-asthenozoospermia group(P = 0.002), while the results of flow cytometry indicated no difference(P = 0.152). In terms of ROS levels, compared with flow cytometry, SSRDM has a stronger ability to distinguish between those two groups, providing a reliable basis for assessment of sperm quality. Another 30 semen samples were used to investigate temperature and temporal variability of SSRDM to ensure the stability and accuracy of this method. Overall, we have developed a method that can quantitatively detect fluorescent substances in sperm at the single-cell level supplying evidence for diagnosis and prognosis of male infertility.展开更多
Objective Endometrial carcinoma(EC)is a prevalent gynecological malignancy characterized by increasing incidence and mortality rates.This underscores the critical need for novel therapeutic targets.One such potential ...Objective Endometrial carcinoma(EC)is a prevalent gynecological malignancy characterized by increasing incidence and mortality rates.This underscores the critical need for novel therapeutic targets.One such potential target is cell division cycle 20(CDC20),which has been implicated in oncogenesis.This study investigated the effect of the CDC20 inhibitor Apcin on EC and elucidated the underlying mechanism involved.Methods The effects of Apcin on EC cell proliferation,apoptosis,and the cell cycle were evaluated using CCK8 assays and flow cytometry.RNA sequencing(RNA-seq)was subsequently conducted to explore the underlying molecular mechanism,and Western blotting and coimmunoprecipitation were subsequently performed to validate the results.Animal studies were performed to evaluate the antitumor effects in vivo.Bioinformatics analysis was also conducted to identify CDC20 as a potential therapeutic target in EC.Results Treatment with Apcin inhibited proliferation and induced apoptosis in EC cells,resulting in cell cycle arrest.Pathways associated with apoptosis and the cell cycle were activated following treatment with Apcin.Notably,Apcin treatment led to the upregulation of the cell cycle regulator p21,which was verified to interact with CDC20 and consequently decrease the expression of downstream cyclins in EC cells.In vivo experiments confirmed that Apcin treatment significantly impeded tumor growth.Higher CDC20 expression was observed in EC tissue than in nonmalignant tissue,and increased CDC20 expression in EC patients was associated with shorter overall survival and progress free interval.Conclusion CDC20 is a novel molecular target in EC,and Apcin could be developed as a candidate antitumor drug for EC treatment.展开更多
BACKGROUND The prevalence of female infertility between the ages of 25 and 44 is 3.5%to 16.7%in developed countries and 6.9%to 9.3%in developing countries.This means that infertility affects one in six couples and is ...BACKGROUND The prevalence of female infertility between the ages of 25 and 44 is 3.5%to 16.7%in developed countries and 6.9%to 9.3%in developing countries.This means that infertility affects one in six couples and is recognized by the World Health Organization as the fifth most serious global disability.The International Committee for Monitoring Assisted Reproductive Technology reported that the global total of babies born as a result of assisted reproductive technology procedures and other advanced fertility treatments is more than 8 million.Advancements in controlled ovarian hyperstimulation procedures led to crucial accomplishments in human fertility treatments.The European Society for Human Reproduction and Embryology guideline on ovarian stimulation gave us valuable evidence-based recommendations to optimize ovarian stimulation in assisted reproductive technology.Conventional ovarian stimulation protocols for in vitro fertilization(IVF)–embryo transfer are based upon the administration of gonadotropins combined with gonadotropin-releasing hormone(GnRH)analogues,either GnRH agonists(GnRHa)or antagonists.The development of ovarian cysts requires the combination of GnRHa and gonadotropins for controlled ovarian hyperstimulation.However,in rare cases patients may develop an ovarian hyper response after administration of GnRHa alone.CASE SUMMARY Here,two case studies were conducted.In the first case,a 33-year-old female diagnosed with polycystic ovary syndrome presented for her first IVF cycle at our reproductive center.Fourteen days after triptorelin acetate was administrated(day 18 of her menstrual cycle),bilateral ovaries presented polycystic manifestations.The patient was given 5000 IU of human chorionic gonadotropin.Twenty-two oocytes were obtained,and eight embryos formed.Two blastospheres were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.In the second case,a 37-year-old woman presented to the reproductive center for her first donor IVF cycle.Fourteen days after GnRHa administration,the transvaginal ultrasound revealed six follicles measuring 17-26 mm in the bilateral ovaries.The patient was given 10000 IU of human chorionic gonadotropin.Three oocytes were obtained,and three embryos formed.Two high-grade embryos were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.CONCLUSION These two special cases provide valuable knowledge through our experience.We hypothesize that oocyte retrieval can be an alternative to cycle cancellation in these conditions.Considering the high progesterone level in most cases of this situation,we advocate freezing embryos after oocyte retrieval rather than fresh embryo transfer.展开更多
Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease dia...Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods.Terminal deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 30-OH end of single-stranded DNA in the absence of a template.In this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP activity.After ALP hydrolyzes the 30-PO_(4) end of the substrate and generates 30-OH,TdT can effectively elongate the 30-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T probes.Endo Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 30-OH end,leading to the generation of exponential fluorescence signal amplification.The substrate for TdT elongation was optimized,and a limit of detection of 4.3×10^(-3) U/L was achieved for ALP by the optimized substrate structure.This method can also detect ALP in the cell lysate of a single cell.This work has potential applications in disease diagnosis and biomedical detection.展开更多
Strand displacement reaction enables the construction of enzyme-free DNA reaction networks,thus has been widely applied to DNA circuit and nanotechnology.It has the characteristics of high efficiency,universality and ...Strand displacement reaction enables the construction of enzyme-free DNA reaction networks,thus has been widely applied to DNA circuit and nanotechnology.It has the characteristics of high efficiency,universality and regulatability.However,the existing regulation tools cannot enable effective control of the reaction sequence,which undoubtedly limits the construction of complex nucleic acid circuits.Herein,we developed a regulation tool,toehold lock,and achieved strict control of reaction sequence without loss of the main reaction signal output.Furthermore,we applied the tool to scenarios such as seesaw circuits,AND/OR logic gates,and entropy-driven circuits,and respectively demonstrated its significant superiority compared to the original method.We believe that the proposed toehold lock has greatly optimized the efficiency of DNA strand displacement-based networks,and we anticipate that the tool will be widely used in multiple fields.展开更多
MicroRNAs(miRNAs)are mediators of the aging process.The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility.Twenty-seven donors were divide...MicroRNAs(miRNAs)are mediators of the aging process.The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility.Twenty-seven donors were divided into three groups by age(Group A,n=8,age:20–30 years;Group B,n=10,age:31–40 years;and Group C,n=9,age:41–55 years)for high-throughput sequencing analysis.Samples from 65 individuals(22,22,and 21 in Groups A,B,and C,respectively)were used for validation by quantitative real-time polymerase chain reaction(qRT-PCR).A total of 2160 miRNAs were detected:1223 were known,937 were newly discovered and unnamed,of which 191 were expressed in all donors.A total of 7,5,and 17 differentially expressed microRNAs(DEMs)were found in Group A vs B,Group B vs C,and Group A vs C comparisons,respectively.Twenty-two miRNAs were statistically correlated with age.Twelve miRNAs were identified as age-associated miRNAs,including hsa-miR-127-3p,mmu-miR-5100_L+2R-1,efu-miR-9226_L-2_1ss22GA,cgr-miR-1260_L+1,hsa-miR-652-3p_R+1,pal-miR-9993a-3p_L+2R-1,hsa-miR-7977_1ss6AG,hsa-miR-106b-3p_R-1,hsa-miR-186-5p,PC-3p-59611_111,hsa-miR-93-3p_R+1,and aeca-mir-8986ap5_1ss1GA.There were 9165 target genes of age-associated miRNAs.Gene Ontology(GO)analysis of the target genes identified revealed enrichment of protein binding,membrane,cell cycle,and so on.The Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of age-related miRNAs for target genes revealed 139 enriched pathways,such as signaling pathways regulating stem cell pluripotency,metabolic pathways,and the Hippo signaling pathway.This suggests that miRNAs play a key role in male fertility changes with increasing age and provides new evidence for the study of the mechanism of age-related male fertility decline.展开更多
Premature Rupture of Membranes (PROM) with the resulting prematurity remains a major public health issue in the Democratic Republic of Congo (DRC). This study aimed to assess expectant management of PPROM before 34 we...Premature Rupture of Membranes (PROM) with the resulting prematurity remains a major public health issue in the Democratic Republic of Congo (DRC). This study aimed to assess expectant management of PPROM before 34 weeks at the university hospital of Kinshasa. We conducted a retrospective analysis of expectantly managed PROM before 34 weeks between January 2008 and December 2018. Maternal and fetal outcomes were collected, and all data were analyzed using the SPSS 23.0 software. Of the 113 patients included in the study, 2.6% were diagnosed with PROM before 34 weeks. We observed prolongation of the pregnancy duration;the median latency period was eight days, and the average gestational age at delivery of 32.85 ± 2.5 weeks. Chorioamnionitis (23%), severe oligoamnios (7%), and acute fetal distress (4%) were complications observed during the latency period. In the postpartum period, endometritis (6.2%), neonatal jaundice (39.8%), anemia (25.7%), ulcerative necrotizing enterocolitis (6.2%), cerebromeningeal hemorrhage (5.3%), and acute respiratory distress syndrome (4.4%) were complications observed. The risk of infection during the latency period was significantly associated with irregular (P = 0.045) or lack (P = 0.006) antenatal care (ANC) attendances and C-Reactive Protein (CRP) results 6 (P = 0.013). The risk of neonatal death was significantly associated to infection during the latency period (P = 0.011), irregular (P = 0.009) or lack of ANC (P = 0.000) attendances, Birth weight g (P = 0.039) as well as Gestational age at birth between 28 to 30 Weeks (S) (P = 0.021). These findings report first-time pregnancy outcomes related to the management of PPROM before 34 weeks in our setting. We found that the conservative attitude adopted allowed the prolongation of pregnancies, reducing the risks associated with prematurity. Nevertheless, attendance in good quality ANC could reduce the frequency of PROM and related adverse outcomes.展开更多
Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivi...Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivity to external conditions.However,achieving enhanced amplification or decision-making necessitates the incorporation of multiple strands,thereby increasing the risk of contamination.Recent advancements have led to the development of CRISPR-Cas-based DNA circuits.These systems aim to simplify the complexity associated with conventional circuits,mitigate contamination risks,and enable more substantial amplification or decision-making capabilities.Here,the review article centers on current strategies of CRISPR-Cas(Cas9,Cas12a,Cas13a)system-assisted circuits in amplification and decisionmaking,and assesses their tendencies and limitations in amplification circuits and decision-making circuits.Furthermore,we discuss the challenges of CRISPR-Cas in circuits and propose prospects that will contribute to constructing more efficient and diverse CRISPR-Cas-based DNA functional circuits.展开更多
文摘Objective:To assess whether personalized embryo transfer guided by endometrial receptivity array(ERA)improves implantation and pregnancy rates in women with implantation failure.Methods:This retrospective case-control study was conducted on women with previous implantation failure.The women were divided into two groups,i.e,women who underwent ERA and those who underwent embryo transfer without ERA testing.ERA was performed using Igenomix.ERA results were interpreted as receptive or non-receptive.Women underwent frozen embryo transfer on the 6th day of progesterone(P+5).The primary outcomes were implantation rate,clinical pregnancy rate,abortion rate,and negative pregnancy rate.Results:This study included 229 women with previous implantation failure,with 154 in the ERA group and 75 in the no ERA group.The mean age of the women of the ERA group was(32.2±4.1)years,and that of the no ERA group was(31.5±4.8)years.Women in the ERA group had a higher implantation rate(60.4%)and clinical pregnancy rate(57.1%)compared to those in the no ERA group(48.0%and 46.7%,respectively).In addition,implantation rate of the nonreceptive ERA group was higher than the no ERA group(65%vs.48%),and clinical pregnancy rate was also higher in the non-receptive ERA group than the no ERA group(65%vs.47%).The abortion rate of the no ERA group was 9%and that of the non-receptive ERA group was 10%.52%no ERA group women and 35%non-receptive ERA group women had negative pregnancy results.Conclusions:Women who have undergone personalised embryo transfer guided by ERA have a higher clinical pregnancy rate than women who have not after previous implantation failure.
基金supported by the Open Research Fund of National Health Commission Key Laboratory of Birth Defects Prevention&Henan Key Laboratory of Population Defects Prevention(ZD202308).
文摘Background:Four-dimensional(4D)ultrasound is increasingly being used for prenatal diagnosis of congenital heart disease(CHD).We aimed to perform a systematic review and meta-analysis to evaluate its diagnostic accuracy for fetal CHD.Methods:This systematic review was conducted in accordance with the PRISMA-DTA guidelines.We systematically searched eight databases for studies published up to July 22,2025.Data were extracted to calculate diagnostic accuracy metrics,study quality was assessed using QUADAS-2,and a bivariate random-effects model was used for the meta-analysis.Results:A total of 49 studies were included,comprising 45 retrospective and 4 prospective studies,which were mainly(91.8%)conducted in China.These studies involved 23,397 fetuses,among which 2115 were diagnosed with congenital heart disease.The pooled sensitivity of 4D ultrasound for diagnosing fetal congenital heart disease was 0.91(95%CI:0.89-0.93),the pooled specificity was 0.98(95%CI:0.97-0.99),and the area under the summary receiver operating characteristic(SROC)curve(AUC)was 0.98(95%CI:0.96-0.99).Although 4D ultrasound technology can be implemented as early as 11 weeks of gestation,its diagnostic sensitivity and specificity reached a superior level and stabilized at an average of 20 weeks of gestation(range 14-28 weeks).Meta-regression indicated that sample size and prior suspicion of fetal CHD were significant contributors to heterogeneity(p<0.05).Conclusion:Four-dimensional ultrasound has high diagnostic efficacy for fetal CHD and is suitable for prenatal screening of fetal CHD,and the diagnostic effect is optimal and stable at an average gestational age of 20 weeks(range 14-28 weeks).
基金supported by the National Natural Science Foundation of China(8200340181972981)+1 种基金the Scientific and Technological Project of Henan Province(222102310150,China)the Open Research Fund of the National Health Commission Key Laboratory of Birth Defects Prevention(NHCKLBDP202504,China).
文摘Preterm birth(PTB),defined as delivery before 37 weeks of gestation,is the most common adverse pregnancy outcome[1].PTB is a global health concern,with an estimated 13.4 million cases in 2020[1],accounting for more than one in 10 births worldwide.Compared to full-term births,PTBs are associated with a higher risk of short-and long-term complications,including bronchopulmonary dysplasia,necrotizing enterocolitis,visual impairment,and cerebral injuries[2].Despite substantial research efforts to prevent PTB,the global PTB rate has shown little improvement over the past decade[1].Therefore,identifying additional risk factors remains a critical goal in preventing PTB.
基金supported by grants from the National Key Research and Development Program of China(2019YFA0802704)the National Natural Science Foundation of China(31771620)+2 种基金the Natural Science Foundation of Chongqing,China(CSTB2022NSCQMSX1424)Research Startup Fund of Southwest University(SWU117064)Open Research Fund of National Health Commission Key Laboratory of Birth Defects Prevention&Henan Key Laboratory of Population Defects Prevention(ZD202302)。
文摘Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(Reiter and Leroux,2017;Wallmeier et al.,2020).To date,over 180 ciliopathy-associated genes have been identified(Reiter and Leroux,2017),yet the underlying mechanisms remain poorly understood.
文摘Objective To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro.The specific mechanism by which microRNA-144-3p is involved in Cu^(2+)-induced damage to the human endometrial epithelial cells(HEECs)was explored.Methods HEECs were cultured in copper-containing culture medium to simulate changes in the endometrium after copper intrauterine device(Cu-IUD)implantation.Reverse transcription quantitative PCR(RT-qPCR)was used to detect the differential expression of miR-144-3p in HEECs after Cu^(2+)treatment.MiRNAs,siRNAs and related inhibitors were used to treat HEECs.The expression levels of related downstream genes were then analyzed by RT-qPCR,Western blotting and immunofluorescence to explore the specific mechanism involved.Results MiR-144-3p was significantly upregulated in the Cu^(2+)-treated HEECs.The expression of P-NF-κB,MMP9,TGF-β3 and P-SMAD3 was significantly decreased in HEECs treated with 10μg/mL Cu^(2+).MiR-144-3p regulated the expression of metallothionein 1A(MT1A)and thrombospondin-1(THBS-1)in Cu^(2+)-treated HEECs.The expression of P-NF-κB can be regulated by MT1A,and an inhibitor of P-NF-κB can significantly reduce the expression of MMP9 in Cu^(2+)-treated HEECs.The expression of TGF-β3 can be regulated by THBS-1,and a TGF-β3 inhibitor can significantly reduce the expression of SMAD3 in Cu^(2+)-treated HEECs.The proliferative capacity of HEECs treated with MMP9 or SMAD3 inhibitors was significantly reduced.Conclusions The increased Cu^(2+)concentration led to the upregulation of miR-144-3p,further reducing the expression levels of its target genes(MT1A and THBS-1),which in turn downregulated the expression of NF-κB,MMP9,TGF-β3 and SMAD3,ultimately leading to increased endometrial cell damage and decreased cell proliferation.
基金financially supported by the Natural Science Foundation of Wuhan City (Chenguang Project) (No.2024040801020331)the Natural Science Foundation of Hubei Province of China (No.2023AFB402)+1 种基金the National Key Research and Development Program of China (No.2023YFE0210200)Interdisciplinary Research Program of HUST。
文摘DNA methylation is an important promising biomarker for cancer diagnosis and monitoring.Therefore,the assessment of DNA methylation levels is helpful for the prognosis and diagnosis of cancer.However,it is still a huge challenge to sensitively and accurately quantify the levels of DNA methylation in clinical sample.In this work,we proposed a protospacer adjacent motif(PAM)-free mediated CRISPR-Cas12a ultra-sensitive and quantitative DNA methylation detection method.Through recognizing the ds DNA with toehold region,CRISPR-Cas12a not only got rid of the limitation of PAM,but also improved its distinction ability for single Cp G site methylation,nearly 5-fold that of conventional PAM-containing ds DNA.We further introduced assist-strand and design an artificial mismatch to greatly improve the ability to distinguish single Cp G methylation site.Our results showed that the discrimination factor was >200.Then,we constructed toe-ds DNA by using “heating and freezing”,which made our method universally applicable and feasible.In addition,we greatly simplified the difficulty of primer design.Our method detected four highly methylated genes acyl carrier protein(ACP),CLV3/ESR-related(CLE),Disabled(DAB) and Homeobox(HOX) with a detection limit of 0.01 % and excellent linearity in DNA methylation standards.Then,we verified the clinical utility of this method in 29 hepatocellular carcinomas,11 ovarian cancers and4 health people.In conclusion,we have successfully constructed a PAM-free CRISPR-Cas12a DNA methylation quantification method,which achieves high congruence in sensitivity,specificity and universality,fully demonstrating its significant clinical application value.
基金This work was funded by National Key R&D Program of China(2018YFC1004300 and 2018YFC1004304)National Natural Foundation of China(No.81871148 and No.818701539).
文摘Male reproductive infections are known to shape the immunological homeostasis of the testes,leading to male infertility.However,the specific pathogenesis of these changes remains poorly understood.Exosomes released in the inflammatory microenvironment are important in communication between the local microenvironment and recipient cells.Here,we aim to identify the immunomodulatory properties of inflammatory testes-derived exosomes(IT-exos)and explore their underlying mechanisms in orchitis.IT-exos were isolated using a uropathogenic Escherichia coli(UPEC)-induced orchitis model and confirmed that IT-exos promoted proinflammatory M1 activation with increasing expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in vitro.We further used small RNA sequencing to identify the differential miRNA profiles in exosomes and primary testicular macrophages(TMs)from normal and UPEC-infected testes,respectively,and identified that miR-155-5p was highly enriched in IT-exos and TMs from inflammatory testes.Further study of bone marrow derived macrophages(BMDMs)transfected with miR-155-5p mimic showed that macrophages polarized to proinflammatory phenotype.In addition,the mice that were administrated IT-exos showed remarkable activation of TM1-like macrophages;however,IT-exos with silencing miR-155-5p showed a decrease in proinflammatory responses.Overall,we demonstrate that miR-155-5p delivered by IT-exos plays an important role in the activation of TM1 in UPEC-induced orchitis.Our study provides a new perspective on the immunological mechanisms underlying inflammation-related male infertility.
基金This work,in part,was supported by grants from the National Natural Science Foundation of China(31801237 to XLW and 82171605 to SQY)the Science Technology and Innovation Commission of Shenzhen Municipality(JCYJ20170818160910316 to SQY).
文摘Mammalian testis exhibits remarkably high transcriptome complexity,and spermatogenesis undergoes two periods of transcriptional cessation.These make the RNA-binding proteins(RBPs)the utmost importance during male germ cell development.Heterogeneous nuclear ribonucleoproteins(hnRNPs)are a large family of RBPs implicated in many steps of RNA processing;however,their roles in spermatogenesis are largely unknown.Here,we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis.Furthermore,we found that most of the detected hnRNP proteins(hnRNPD,hnRNPK,hnRNPQ,hnRNPU,and hnRNPUL1)display the highest signals in the nuclei of pachytene spermatocytes,round spermatids,and Sertoli cells,whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids.The expression of these hnRNP proteins showed both similarities and specificity,suggesting their diverse roles in spermatogenesis.
文摘Objective:To investigate the association between age and semen parameters among male partners of subfertile couples.Methods:This retrospective study analyzed the semen of 1523 infertile men aged 26 to 50 years.Data were extracted from GarbhaGudi IVF Centre database from January 2019 to September 2020.The basic semen parameters were interpreted according to the WHO manual 2021,6th edition.Semen parameters in different age groups were compared.Results:Total and progressive motile sperms were significantly higher in the age group of 26-30 years compared to other age groups(P<0.05).Normal sperm count was significantly higher in the age group of 26-30 years compared to the age groups of 41-45 years and>46 years(P=0.001).However,sperm head defects,neck and midpiece defects,tail defects,and cytoplasmic droplets showed statistically insignificant difference in all the age groups(P>0.05).Semen viscosity showed no statistical difference in all the age groups compared to the reference age group of 26 to 30 years.Conclusions:Higher age can lead to a significant decrease in normal sperms and motility in subfertile men.Hence,male partner age should be considered as one of the major determining factors for reproductive outcomes.
基金the National Natural Science Foundation of China (Nos. 82172372 and 21904045)COVID-19 Pneumonia Emergency Scientific Research Special Fund of Wuhan (No. EX20D03)the Fundamental Research Funds for the Central Universities (Nos. 2019kfy XJJS169 and 2021yjs CXCY127)。
文摘DNA-functionalized gold nanoparticles are one of the most versatile bionanomaterials for biomedical and clinical diagnosis. Herein, we discovered that the performance of DNAzyme cleaving the substrate is highly related to its length. This intriguing phenomenon only appears at the interfaces of DNAfunctionalized gold nanoparticles. We systematically investigated the causes of this phenomenon. We conjectured that the DNAzyme with extended nucleotides that do not match its substrate strand is vulnerable to non-specific adsorption, electrostatic repulsion, and steric hindrance. Based on our improved understanding of this phenomenon, we have successfully developed a highly sensitive and specific amplifiable biosensor to detect human apurinic/apyrimidinic endonuclease 1.
基金financially supported by the National Natural Science Foundation of China (Nos.21904045,21705053 and 81871732)the Fundamental Research Funds for the Central Universities (No.2019kfyXJJS169)National Training Program of Innovation and Entrepreneurship for Undergraduates (No. 201910487087)。
文摘We presented a low-abundance mutation detection method with lambda exonuclease and DNA threeway junction structure.The assistant strand in the DNA three-way junction structure could regulate the reaction system from the kinetics and thermodyna mics aspects.The optimization of the assista nt strand helps to improve the selectivity of the mutant-type DNA to the wild-type DNA about 35 times.Moreover,the cost of the optimization process could be saved by about 90%.The method was applied to the detection of a human ovarian cancer-related gene mutation BRCA1(rs1799949,c.2082 C>T).The limit of detection to the mutation abundance in the DNA three-way junction structure system(0.2%) was one order lower compared with that in the double-stranded DNA structure system(2%).The mutation abundance in different standard samples was quantitively measured,and the results were consistent with the initial abundance in the standard samples.
基金supported by the National Natural Foundation of China(81871148)the Open Fund of NHC Key Laboratory of Birth Defects Prevention,Henan Key Laboratory of Population Defects Prevention(Henan Institute of Reproduction Health Science and Technology,ZD202201)the Fundamental Research Funds for the Central Universities,HUST(2023JYCXJJ062 and YCJJ202201050).
文摘Infections and inflammatory reactions in the male genital tract are the leading causes of male infertility with a prevalence of 6%-10%,primarily affecting testicular and epididymal function and ultimately compromising sperm quality.However,most infertile patients with genital infection/inflammation are asymptomatic and easily overlooked.Traditional indicators,including white blood cells,elastase,and other components in semen,can reflect inflammation of the genital tract,but there is still a lack of a uniform standard method of detection.Therefore,it is necessary to explore reliable markers in semen that reflect the inflammatory status of the genital tract.Using the experimental autoimmune orchitis(EAO)model to simulate noninfectious chronic orchitis,we successfully collected ejaculated seminal fluid from EAO rats using optimized electrical stimulation devices.Proteomic analysis was performed using isobaric tags for relative and absolute quantification(iTRAQ).Compared to the control group,55 upregulated and 105 downregulated proteins were identified in seminal plasma samples from the EAO group.In a preliminary screening,the inflammation-related protein S100A8/A9 was upregulated.We further verified that S100A8/A9 was increased in seminal plasma and highly expressed in testicular macrophages of the EAO model.In patients with oligoasthenospermia and genital tract infections,we also found that S100A8/A9 levels were remarkably increased in seminal plasma and testicular macrophages.S100A8/A9 in semen may be a potential biomarker for chronic genital inflammation.Our study provides a new potential biomarker for early diagnosis and further understanding of male infertility caused by genital inflammation.
基金supported by the National Natural Science Foundation of China (Nos. 21705053 and 81871732)the Open Foundation of NHC Key Laboratory of Birth Defects Prevention (No. ZD202104)the Open Foundation of Key Laboratory of Birth Regulation and Control Technology of National Health Commission of China, Maternal Child Health Care Hospital of Shandong Province。
文摘Sperm damage caused by reactive oxygen species(ROS) is one of the main causes of male infertility.Therefore, the level of ROS in sperm is an important indicator for the diagnosis and prognosis of male infertility. Herein, we constructed a single sperm ROS detection method(SSRDM) with an optical microprobe fabricated via focused ion beam process. The micro-probe is used to separately excite fluorescence in the sperm and the area around the sperm after ROS staining, and the difference in fluorescence values can reflect the level of ROS in the sperm. We collected 102 semen samples and 72 of them were divided into asthenozoospermia and non-asthenozoospermia groups. SSRDM and flow cytometry were used to detect the ROS levels of the two groups. The results of SSRDM showed that the ROS levels of asthenozoospermia group were higher than that of non-asthenozoospermia group(P = 0.002), while the results of flow cytometry indicated no difference(P = 0.152). In terms of ROS levels, compared with flow cytometry, SSRDM has a stronger ability to distinguish between those two groups, providing a reliable basis for assessment of sperm quality. Another 30 semen samples were used to investigate temperature and temporal variability of SSRDM to ensure the stability and accuracy of this method. Overall, we have developed a method that can quantitatively detect fluorescent substances in sperm at the single-cell level supplying evidence for diagnosis and prognosis of male infertility.
文摘Objective Endometrial carcinoma(EC)is a prevalent gynecological malignancy characterized by increasing incidence and mortality rates.This underscores the critical need for novel therapeutic targets.One such potential target is cell division cycle 20(CDC20),which has been implicated in oncogenesis.This study investigated the effect of the CDC20 inhibitor Apcin on EC and elucidated the underlying mechanism involved.Methods The effects of Apcin on EC cell proliferation,apoptosis,and the cell cycle were evaluated using CCK8 assays and flow cytometry.RNA sequencing(RNA-seq)was subsequently conducted to explore the underlying molecular mechanism,and Western blotting and coimmunoprecipitation were subsequently performed to validate the results.Animal studies were performed to evaluate the antitumor effects in vivo.Bioinformatics analysis was also conducted to identify CDC20 as a potential therapeutic target in EC.Results Treatment with Apcin inhibited proliferation and induced apoptosis in EC cells,resulting in cell cycle arrest.Pathways associated with apoptosis and the cell cycle were activated following treatment with Apcin.Notably,Apcin treatment led to the upregulation of the cell cycle regulator p21,which was verified to interact with CDC20 and consequently decrease the expression of downstream cyclins in EC cells.In vivo experiments confirmed that Apcin treatment significantly impeded tumor growth.Higher CDC20 expression was observed in EC tissue than in nonmalignant tissue,and increased CDC20 expression in EC patients was associated with shorter overall survival and progress free interval.Conclusion CDC20 is a novel molecular target in EC,and Apcin could be developed as a candidate antitumor drug for EC treatment.
文摘BACKGROUND The prevalence of female infertility between the ages of 25 and 44 is 3.5%to 16.7%in developed countries and 6.9%to 9.3%in developing countries.This means that infertility affects one in six couples and is recognized by the World Health Organization as the fifth most serious global disability.The International Committee for Monitoring Assisted Reproductive Technology reported that the global total of babies born as a result of assisted reproductive technology procedures and other advanced fertility treatments is more than 8 million.Advancements in controlled ovarian hyperstimulation procedures led to crucial accomplishments in human fertility treatments.The European Society for Human Reproduction and Embryology guideline on ovarian stimulation gave us valuable evidence-based recommendations to optimize ovarian stimulation in assisted reproductive technology.Conventional ovarian stimulation protocols for in vitro fertilization(IVF)–embryo transfer are based upon the administration of gonadotropins combined with gonadotropin-releasing hormone(GnRH)analogues,either GnRH agonists(GnRHa)or antagonists.The development of ovarian cysts requires the combination of GnRHa and gonadotropins for controlled ovarian hyperstimulation.However,in rare cases patients may develop an ovarian hyper response after administration of GnRHa alone.CASE SUMMARY Here,two case studies were conducted.In the first case,a 33-year-old female diagnosed with polycystic ovary syndrome presented for her first IVF cycle at our reproductive center.Fourteen days after triptorelin acetate was administrated(day 18 of her menstrual cycle),bilateral ovaries presented polycystic manifestations.The patient was given 5000 IU of human chorionic gonadotropin.Twenty-two oocytes were obtained,and eight embryos formed.Two blastospheres were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.In the second case,a 37-year-old woman presented to the reproductive center for her first donor IVF cycle.Fourteen days after GnRHa administration,the transvaginal ultrasound revealed six follicles measuring 17-26 mm in the bilateral ovaries.The patient was given 10000 IU of human chorionic gonadotropin.Three oocytes were obtained,and three embryos formed.Two high-grade embryos were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.CONCLUSION These two special cases provide valuable knowledge through our experience.We hypothesize that oocyte retrieval can be an alternative to cycle cancellation in these conditions.Considering the high progesterone level in most cases of this situation,we advocate freezing embryos after oocyte retrieval rather than fresh embryo transfer.
基金financially supported by the National Natural Science Foundation of China(Grant No.:21904045)the Fundamental Research Funds for the Central Universities(HUST:Grant No.:2019kfyXJJS169)Training Program of Innovation and Entrepreneurship for Undergraduates of Hubei Province(Grant No.:S202010487225).
文摘Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods.Terminal deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 30-OH end of single-stranded DNA in the absence of a template.In this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP activity.After ALP hydrolyzes the 30-PO_(4) end of the substrate and generates 30-OH,TdT can effectively elongate the 30-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T probes.Endo Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 30-OH end,leading to the generation of exponential fluorescence signal amplification.The substrate for TdT elongation was optimized,and a limit of detection of 4.3×10^(-3) U/L was achieved for ALP by the optimized substrate structure.This method can also detect ALP in the cell lysate of a single cell.This work has potential applications in disease diagnosis and biomedical detection.
基金the financial support from the National Key Research and Development Program of China(No.2021YFC2701402)the Open Research Fund of State Key Laboratory of Bioelectronics,Southeast University(No.Sklb2021-k06)+2 种基金the Open Foundation of NHC Key Laboratory of Birth Defect for Research and Prevention(Hunan Provincial Maternal and Child Health Care Hospital)(No.KF2020007)Hunan Province Assisted Reproduction and Regenerative Medicine Clinical Demonstration Center Funded Project(No.2020SK4019)the Open Foundation of Translational Medicine National Science and Technology Infrastructure(Shanghai)(No.TMSK-2021-141)。
文摘Strand displacement reaction enables the construction of enzyme-free DNA reaction networks,thus has been widely applied to DNA circuit and nanotechnology.It has the characteristics of high efficiency,universality and regulatability.However,the existing regulation tools cannot enable effective control of the reaction sequence,which undoubtedly limits the construction of complex nucleic acid circuits.Herein,we developed a regulation tool,toehold lock,and achieved strict control of reaction sequence without loss of the main reaction signal output.Furthermore,we applied the tool to scenarios such as seesaw circuits,AND/OR logic gates,and entropy-driven circuits,and respectively demonstrated its significant superiority compared to the original method.We believe that the proposed toehold lock has greatly optimized the efficiency of DNA strand displacement-based networks,and we anticipate that the tool will be widely used in multiple fields.
基金This work was supported by the Central Public Interest Scientific Institution Basal Research Fund of National Research Institute for Family Planning(No.2021GJZ09)the CAMS Innovation Fund for Medical Sciences(CIFMS,No.2018-I2M-1-004)。
文摘MicroRNAs(miRNAs)are mediators of the aging process.The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility.Twenty-seven donors were divided into three groups by age(Group A,n=8,age:20–30 years;Group B,n=10,age:31–40 years;and Group C,n=9,age:41–55 years)for high-throughput sequencing analysis.Samples from 65 individuals(22,22,and 21 in Groups A,B,and C,respectively)were used for validation by quantitative real-time polymerase chain reaction(qRT-PCR).A total of 2160 miRNAs were detected:1223 were known,937 were newly discovered and unnamed,of which 191 were expressed in all donors.A total of 7,5,and 17 differentially expressed microRNAs(DEMs)were found in Group A vs B,Group B vs C,and Group A vs C comparisons,respectively.Twenty-two miRNAs were statistically correlated with age.Twelve miRNAs were identified as age-associated miRNAs,including hsa-miR-127-3p,mmu-miR-5100_L+2R-1,efu-miR-9226_L-2_1ss22GA,cgr-miR-1260_L+1,hsa-miR-652-3p_R+1,pal-miR-9993a-3p_L+2R-1,hsa-miR-7977_1ss6AG,hsa-miR-106b-3p_R-1,hsa-miR-186-5p,PC-3p-59611_111,hsa-miR-93-3p_R+1,and aeca-mir-8986ap5_1ss1GA.There were 9165 target genes of age-associated miRNAs.Gene Ontology(GO)analysis of the target genes identified revealed enrichment of protein binding,membrane,cell cycle,and so on.The Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of age-related miRNAs for target genes revealed 139 enriched pathways,such as signaling pathways regulating stem cell pluripotency,metabolic pathways,and the Hippo signaling pathway.This suggests that miRNAs play a key role in male fertility changes with increasing age and provides new evidence for the study of the mechanism of age-related male fertility decline.
文摘Premature Rupture of Membranes (PROM) with the resulting prematurity remains a major public health issue in the Democratic Republic of Congo (DRC). This study aimed to assess expectant management of PPROM before 34 weeks at the university hospital of Kinshasa. We conducted a retrospective analysis of expectantly managed PROM before 34 weeks between January 2008 and December 2018. Maternal and fetal outcomes were collected, and all data were analyzed using the SPSS 23.0 software. Of the 113 patients included in the study, 2.6% were diagnosed with PROM before 34 weeks. We observed prolongation of the pregnancy duration;the median latency period was eight days, and the average gestational age at delivery of 32.85 ± 2.5 weeks. Chorioamnionitis (23%), severe oligoamnios (7%), and acute fetal distress (4%) were complications observed during the latency period. In the postpartum period, endometritis (6.2%), neonatal jaundice (39.8%), anemia (25.7%), ulcerative necrotizing enterocolitis (6.2%), cerebromeningeal hemorrhage (5.3%), and acute respiratory distress syndrome (4.4%) were complications observed. The risk of infection during the latency period was significantly associated with irregular (P = 0.045) or lack (P = 0.006) antenatal care (ANC) attendances and C-Reactive Protein (CRP) results 6 (P = 0.013). The risk of neonatal death was significantly associated to infection during the latency period (P = 0.011), irregular (P = 0.009) or lack of ANC (P = 0.000) attendances, Birth weight g (P = 0.039) as well as Gestational age at birth between 28 to 30 Weeks (S) (P = 0.021). These findings report first-time pregnancy outcomes related to the management of PPROM before 34 weeks in our setting. We found that the conservative attitude adopted allowed the prolongation of pregnancies, reducing the risks associated with prematurity. Nevertheless, attendance in good quality ANC could reduce the frequency of PROM and related adverse outcomes.
基金financially supported by the National Natural Science Foundation of China (Nos. 82172372 and 82260290)the Opening Research Fund of State Key Laboratory of Digital Medical Engineering (No. 2023-M04)
文摘Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivity to external conditions.However,achieving enhanced amplification or decision-making necessitates the incorporation of multiple strands,thereby increasing the risk of contamination.Recent advancements have led to the development of CRISPR-Cas-based DNA circuits.These systems aim to simplify the complexity associated with conventional circuits,mitigate contamination risks,and enable more substantial amplification or decision-making capabilities.Here,the review article centers on current strategies of CRISPR-Cas(Cas9,Cas12a,Cas13a)system-assisted circuits in amplification and decisionmaking,and assesses their tendencies and limitations in amplification circuits and decision-making circuits.Furthermore,we discuss the challenges of CRISPR-Cas in circuits and propose prospects that will contribute to constructing more efficient and diverse CRISPR-Cas-based DNA functional circuits.
