Background To validate the clinical efficacy of non-invasive prenatal diagnosis (NIPD) for spinal muscular atrophy (SMA) in the first trimester and extend its applicability to families without probands.Method From December 2020 to October 2024, 288 high-risk pregnancies were recruited prospectively, with 81 qualifying for NIPD after genetic counseling. Among the eligible cases, parent-based haplotypes were successfully constructed in 75 families (92.6%), while grandparent-based haplotype reconstruction was performed for the remaining 6 cases (7.4%) where proband samples were unavailable. Through targeted sequencing of the SMN1/SMN2 gene and flanking informative SNPs in maternal plasma, fetal haplotypes were inferred by analyzing dosage changes in cell-free DNA (cfDNA) using Bayes factor. All NIPD results were subsequently validated through invasive diagnostic procedures (chorionic villus sampling or amniocentesis).Results The haplotypes were successfully constructed in 81 families through parents or grandparents of the identified variant carriers. 76 families (93.8%) successfully obtained NIPD results, among which the earliest gestational week for successful NIPD was 7+3 weeks, with a minimum fetal fraction of 1.9%. 5 cases were classified ‘no call’ results due to pathogenic variant-adjacent recombination events (2/5), insufficient or unevenly distributed informative SNPs (2/5), and subthreshold fetal fraction (1/5). The average gestational age of NIPD blood drawing is 9 weeks. Validation test showed the NIPD results accuracy was 100%.Conclusion This study demonstrates the clinical feasibility of grandparent-assisted haplotype construction for SMA families without probands and enables accurate early prenatal diagnosis of SMA in first-trimester pregnancies.

Noninvasive prenatal testing for single-gene disorders faces significant challenges due to low fetal DNA abundance in maternal blood, particularly in early gestation. Although next-generation sequencing (NGS) is widely used, it frequently encounters false-negative outcomes and high costs when detecting low-abundance fetal variants. To address these limitations, we developed a highly sensitive and cost-effective analytical method utilizing high-resolution tandem mass spectrometry, capable of accurately detecting single nucleotide variants (SNVs) as early as 5-8 weeks gestation. It employs gas-phase enrichment combined with fragment mass spectral analysis for precise isolation and identification of fetal DNA amidst predominant maternal DNA. Analytical validation with synthetic DNA and cultured cells confirmed exceptional sensitivity, achieving single copy level detection despite significant interference. Clinically, our method accurately detected paternally inherited SNVs in 134 cell-free fetal DNA samples collected at 5-8 weeks gestation, achieving 100% accuracy. Additionally, it reliably identified pathogenic SNVs linked to phenylketonuria from 12 samples collected at 11-13 weeks gestation. This approach significantly reduces operational costs, enhances sensitivity, and eliminates false-negative results compared to traditional NGS-based methods.

INTRODUCTION:To evaluate the prognostic significance of double-expressor phenotype (DEP), International Prognostic Index (IPI) score, and treatment-related factors in patients with primary testicular diffuse large B-cell lymphoma (PT-DLBCL), and to explore a risk stratification model integrating DEP and IPI. PATIENTS AND METHODS:In this multicenter retrospective cohort study, 67 patients with PT-DLBCL treated at 3 tertiary hospitals in China between February 2013 and December 2024 were included. Clinical, pathological, and treatment-related data were collected from electronic medical records. Overall survival (OS) and progression-free survival (PFS) were analyzed using the Kaplan-Meier method. Univariable and multivariable Cox regression analyses were performed to identify prognostic factors. Based on the independent predictors for OS, an exploratory DEP-IPI risk model was constructed. RESULTS:Among the 67 patients, DEP positivity was associated with significantly inferior OS and PFS, while patients with an IPI score ≥ 3 had significantly worse OS. In multivariable analysis, IPI score ≥ 3 (adjusted hazard ratio = 13.61, P = .042) and DEP positivity (adjusted hazard ratio =13.52,P = .005) remained independent adverse prognostic factors for OS. By contrast, no variable retained independent significance for PFS in the multivariable model. Contralateral testicular radiotherapy was associated with improved OS in Kaplan-Meier analysis but was not significant in Cox regression. Based on IPI and DEP status, patients were stratified into low-, intermediate-, and high-risk groups, which showed significantly different OS (P < .001). Treatment patterns did not differ significantly among the 3 risk groups. CONCLUSION:IPI score ≥ 3 and DEP positivity are independent adverse prognostic factors for OS in PT-DLBCL, whereas no variable retained independent significance for PFS in multivariable analysis. An exploratory DEP-IPI model may help identify clinically distinct risk groups and provide additional prognostic stratification. Further large-scale prospective studies are needed to validate its clinical utility.

Zinc is an essential trace element crucial for human health. The prostate contains the highest concentration of zinc in the male body, making it the organ most affected by zinc disturbances. Zinc plays a critical role in maintaining normal prostate structure and function by modulating energy metabolism and secretory processes in prostate cells, regulating androgen balance, and exerting antibacterial effects. Disturbances in zinc homeostasis are closely associated with the pathogenesis of common prostatic diseases, including prostatitis, benign prostatic hyperplasia (BPH), and prostate cancer (PCa). Consequently, zinc supplementation has emerged as a potential strategy for managing certain prostatic diseases. However, its efficacy and safety remain controversial. This review presents a comprehensive overview of zinc metabolism and mechanisms of action, with a focus on its role in the pathogenesis of common prostatic diseases, and analyzes recent advancements and current challenges in the clinical application of zinc supplementations.

BackgroundNext-generation sequencing (NGS) facilitates simultaneous carrier screening for multiple single-gene disorders. However, conventional NGS methods struggle to detect complex variants, such as F8 inversions, CYP21A2 variations, and single-exon copy number variations (CNVs), resulting in residual risk.MethodsWe developed and validated MLDP-AS (Multiplex Long-Distance PCR followed by Amplicon Sequencing), a novel NGS assay, to screen for pathogenic variants in ten prevalent single-gene disorders in the Chinese population, including alpha- and beta-thalassemia, non-syndromic hearing loss, spinal muscular atrophy, Duchenne muscular dystrophy, phenylketonuria, 21-hydroxylase deficiency, Wilson disease, methylmalonic acidemia, and Hemophilia A. MLDP-AS detects routine variants and technically challenging types, such as F8 intron 22 inversions, CYP21A2 variations, and single-exon CNVs, in a single test. The assay was optimized using positive clinical samples, with sensitivity validated against gold-standard methods. Clinical applicability was evaluated through a prospective study of couples planning or undergoing pregnancy, with positive results confirmed by gold-standard techniques.ResultsMLDP-AS achieved 100% sensitivity, identifying all 255 pathogenic variants in known positive samples, including 36 technically challenging variants. In a prospective trial involving 5,209 individuals, carriers for all targeted disorders were detected, with an overall carrier rate of 22.15%. Thirty-four couples (1.09%) were identified as at-risk, spanning seven of the ten diseases. A total of 289 pathogenic variants were detected 1,290 times, including 169 technically challenging variants. The assay demonstrated a positive predictive value of 99.7% compared to gold-standard methods. MLDP-AS is rapid and cost-effective, completing testing within three days at a cost under $25.ConclusionsMLDP-AS integrates detection of multiple complex variants into a single, comprehensive assay, overcoming limitations of conventional NGS. It significantly enhances variant detection and provides an efficient, cost-effective tool for carrier screening in the Chinese population.

The role of nonsteroidal anti-inflammatory drugs (NSAIDs) in urological carcinogenesis remains controversial, with epidemiological studies reporting inconsistent findings across cancer sites and drug classes. We performed an updated systematic review and meta-analysis to evaluate the associations of overall NSAID use and specific NSAID classes with the risk of kidney, prostate, and bladder cancers. We systematically searched PubMed, Web of Science, EMBASE, and the Cochrane Library for epidemiological studies published up to May 1, 2025. Pooled relative effect estimates and 95

BackgroundCongenital adrenal hyperplasia (CAH) is a common metabolic genetic disease. Early diagnosis and intervention are crucial to improve the prognosis. Noninvasive prenatal diagnosis (NIPD) is an early, safe, and accurate method. This study aimed to evaluate the NIPD of CAH while guiding individualized intrauterine treatment.MethodsTwenty families with a 25% risk of having a baby with 21-hydroxylase deficiency (21-OHD) were included. Haplotypes were constructed based on targeted sequencing and family linkage analysis. Relative haplotype dosage (RHDO) combined with Bayes factor was used to infer fetal genotypes. Invasive prenatal diagnosis was performed to verify the reliability of NIPD. For affected-female fetuses, intrauterine treatment was applied until delivery.ResultsIn 20 families, NIPD successfully identified one female-affected fetus, four male-affected fetuses, nine heterozygotes, and five normal fetuses. The first-pass success rate of NIPD was 90% (18/20), the reporting rate was 95% (19/20), and the accuracy was 100% (19/19). Individualized intrauterine treatment avoided 88.9% (8/9) of unnecessary treatment of unaffected female fetuses. Moreover, no significant virilization was observed in the newborn of CAH16, which underwent intrauterine treatment.ConclusionNIPD has far-reaching implications for the early treatment and clinical management of pregnancy in families with 21-OHD.

Abstract Objective Spinal muscular atrophy (SMA) is a motor neuron disorder encompassing 5q and non-5q forms, causing muscle weakness and atrophy due to spinal cord cell degeneration. Understanding its genetic basis is crucial for genetic counseling and personalized treatment options. Methods This study retrospectively analyzed families of patients suspected of SMA at our institution from February 2006 to March 2024. Various molecular techniques, including multiplex ligation-dependent probe amplification analysis, long-range polymerase chain reaction (PCR) combined with nested PCR, Sanger sequencing, and whole-exome sequencing were employed to establish a thorough genetic variant profile in 680 Chinese pedigrees with clinically suspected SMA. Results Out of 680 families suspected of having SMA, 675 exhibited mutations in the SMN1 gene, while three families were linked to mutations in the IGHMBP2 gene. One family exhibited a genetic variation in the NEB gene, and another family exhibited a variation in the SCO2 gene. Among the families with mutations in the SMN1 gene, 645 families exhibited either E7‒E8 or E7 homozygous deletion. Some families displayed E7‒8 heterozygous deletions along with other mutations, such as E1 or E1‒6 heterozygote deletion and point mutations. Furthermore, one family demonstrated a compound-heterozygous double mutation, while another carried a type “2 + 0” mutation alongside a point mutation. Conclusions This study comprehensively analyzed the genetics of suspected familial SMA cases in the Chinese population, providing insights into the molecular genetic mechanisms of SMA and the utility of various detection techniques. The findings revealed important implications for genetic counseling, prenatal diagnosis, and targeted therapies in clinical practice.

Abstract Background Noninvasive prenatal diagnosis (NIPD) has been proven feasible for non-syndromic hearing loss (NSHL) in singleton pregnancies. However, previous research is limited to the second trimester and the application in twin pregnancies is blank. Here we provide a novel algorithmic approach to assess singleton and twin pregnancies in the first trimester. Methods A 324.614 kb capture panel was designed to selectively enrich target regions. Parental haplotypes were constructed by target sequencing of blood samples from the parents and the proband. Then single nucleotide polymorphisms (SNP) within target regions were classified into four and six categories in singleton and twin pregnancy, respectively. Combining relative haplotype dosage change (RHDO) and the Bayes factor (BF), fetal fraction (FF) and fetal genotype were deduced in singleton and twin pregnancies. The pregnant women’s NIPD results were validated by invasive prenatal diagnosis and Sanger sequencing. Results Sixteen women with singleton pregnancies and one woman with a twin pregnancy were recruited. Among the 16 singleton pregnancies, NIPD was successfully applied in 15 families and the coincidence rate with invasive prenatal diagnosis was 100% (15/15). Only one family NIPD result is “no call” because the imbalance distribution of SNP sites makes it difficult to estimate recombination events. Most (13/15) of pregnant women were diagnosed in the first trimester and the earliest gestation week was the 7th week. The twin pregnancy was a dichorionic diamniotic twin (DCDA). NIPD confirmed one fetus is affected, and another is a carrier with c.299_300delAT of GJB2 gene. Conclusion This study represents the pioneering evidence in the field, demonstrating the feasibility of NIPD for NSHL in twin pregnancies. Moreover, it provides a novel and advanced diagnostic approach for families at high risk of NSHL during pregnancy, offering earlier detection, enhanced safety, and improved accuracy.

Prostate cancer (PCa) remains the most prevalent malignancy among men, with the inevitable emergence of castration-resistant prostate cancer (CRPC) presenting the greatest challenge. Accumulating evidence has confirmed that the overexpression of cAMP response element-binding protein (CREB)-binding protein (CBP) and E1A-binding protein (p300), two highly homologous transcriptional coactivators, plays a crucial role in the development of PCa and its progression to CRPC, thereby making them prominent therapeutic targets for all types of PCa. In this review, we systematically discuss the structure and function of CBP/p300 and elucidate the detailed mechanisms by which CBP/p300 promote prostate carcinogenesis and development. Specifically, CBP/p300 facilitate prostate carcinogenesis by acetylating specific lysine residues on essential transcription factors involved in androgen receptor (AR) signaling, canonical Wnt signaling, p53 signaling, as well as other pathways such as PI3K/AKT and MAPK signaling. Additionally, they contribute to tumor immunosuppression and adaptive resistance to programmed death ligand 1 (PD-L1) blockade treatment by inducing the expression and secretion of the PD-L1 protein. Furthermore, we explore the latest advances in the use of various inhibitors targeting different domains of CBP/p300 and proteolysis-targeting chimeras (PROTAC) degraders in PCa. We propose that combing CBP/p300 inhibitors or degraders with current anti-PCa therapies, including androgen deprivation therapy (ADT), chemotherapy, and immunotherapy, holds potential to overcome the challenges in treating advanced PCa and improve clinical outcomes for all PCa patients.

Aflatoxins, particularly aflatoxin B1 (AFB1), are widely present in the environment and pose a significant risk to food safety. However, no specific drugs are available for AFB1-induced toxic liver injury. Ganfule (GFL) is a traditional Chinese medicine (TCM) formula approved in China for the adjunctive treatment of advanced liver cancer. Nonetheless, its effects on AFB1-induced toxic liver injury remain unclear. In this study, cell and animal experiments have shown that GFL activates the Kelch like ECH associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant pathway in liver or hepatocytes. This leads to an increase in superoxide dismutase (SOD) activity and glutathione (GSH) level, a decrease in reactive oxygen species (ROS) and malondialdehyde (MDA) levels, and inhibition of AFB1-induced oxidative stress. GFL also reduces the levels of inflammatory factors and fibrosis-related genes by inhibiting the interleukin 6 (IL6)/signal transducer and activator of transcription 3 (STAT3) pathway. Furthermore, GFL suppresses the expression of pro-apoptotic genes while enhancing the expression of anti-apoptotic genes, effectively preventing AFB1-induced hepatocyte apoptosis and improving liver function in mice. Network pharmacology predictions, and transcriptome sequencing confirms that GFL exerts its therapeutic effect through the regulation of the aforementioned pathways. Molecular docking analysis, molecular dynamics simulation, and in vitro experiments suggest that quercetin (QCT) mediates its effects through specifically binding to IL6 and STAT3, resulting in the subsequent inhibition of the IL6/STAT3 signaling pathway. This study elucidates the pharmacological mechanism of GFL in treating AFB1-induced toxic liver injury, providing theoretical evidence for its clinical application.

BACKGROUND:Ectopic prostate tissue is a rare clinical lesion that typically originates within the male urogenital tract. It is even more uncommon to find outside the urogenital tract or in female patients. The clinical symptoms of ectopic prostate tissue vary depending on the location of the disease. CASE PRESENTATION:We present the case of a 56-year-old Chinese male of Asian descent who presented with a space-occupying left ureter during a physical examination but did not exhibit any specific clinical symptoms. Preoperative urinary tract color ultrasound and pelvic magnetic resonance imaging examinations revealed nodular, abnormal signal shadows in the lower segment of the left ureter, necessitating further investigation into their nature. The initial preoperative diagnosis leaned towards a ureteral tumor owing to the uncertain nature of the lesion. During laparoscopic left ureteral exploration, rapid pathological sections indicated the presence of mature prostate gland tissue within the affected area. Subsequent laparoscopic left ureteral bladder replantation was performed, and postoperative pathology, along with immunohistochemistry results, confirmed the presence of ureteral ectopic prostate tissue. A comprehensive review of relevant literature, both domestic and foreign, was conducted to analyze the location of the ectopic prostate tissue, clinical characteristics, and various diagnosis and treatment options for this condition. CONCLUSION:Ureteral ectopic prostate tissue is a rare disease that can be challenging to diagnose and treat accurately. Surgical removal of the affected tissue and subsequent histopathological examination are crucial steps to confirm the diagnosis. While the short-term prognosis for this condition is generally positive, long-term follow-up is necessary for optimal management.

Cancer is one of the most significant public health challenges in the new millennium, and complex mechanisms are at work to contribute to its pathogenesis and progression. The Wnt signaling pathways, which are crucial conserved cascades involved in embryological development and tissue homeostasis, and mitochondria, the intracellular powerhouses responsible for energy production, calcium and iron homeostasis, as well as mitochondrial apoptosis in eukaryotic cells, have their own mechanisms regulating these pathological processes. In the past decade, accumulating evidence has indicated that Wnt signaling pathways directly regulate mitochondrial biogenesis and function under physiological and pathological conditions. In this review, we systemically summarize the current understanding of how Wnt signaling pathways, particularly the canonical Wnt cascade, regulate mitochondrial fission, respiration, metabolism, and mitochondrial-dependent apoptosis in cancer. In addition, we discuss recent advancements in the research of anticancer agents and related pharmacological mechanisms targeting the signaling transduction of canonical Wnt pathway and/or mitochondrial function. We believe that the combined use of pharmaceuticals targeting Wnt signaling and/or mitochondria with conventional therapies, immunotherapy and targeted therapy based on accurate molecular pathological diagnosis will undoubtedly be the future mainstream direction of personalized cancer treatment, which could benefit more cancer patients.

The study is to explore the feasibility and value of SNP-based noninvasive prenatal diagnosis (NIPD) for facioscapulohumeral muscular dystrophy type 1 (FSHD1) in early pregnancy weeks. We prospectively collected seven FSHD1 families, with an average gestational age of 8+6. Among these seven couples, there were three affected FSHD1 mothers and four affected fathers. A multiplex-PCR panel comprising 402 amplicons was designed to selective enrich for highly heterozygous SNPs upstream of the DUX4 gene. Risk haplotype was constructed based on familial linkage analysis. Fetal genotypes were accurately inferred through relative haplotype dosage analysis using Bayes Factor. All tests were successfully completed in a single attempt, and no recombination events were detected. NIPD results were provided within a week, which is 4 weeks earlier than karyomapping and 7 weeks earlier than Bionano single-molecule optical mapping (BOM). Ultimately, five FSHD1 fetuses and two normal fetuses were successfully identified, with a 100% concordance rate with karyomapping and BOM. Therefore, SNP-based NIPD for FSHD1 was demonstrated to be feasible and accurate in early weeks of gestation, although the risk of recombination events cannot be completely eliminated. In the future, testing of more cases is still necessary to fully determine the clinical utility.

Objective: To investigate the clinicopathological, immunophenotypic and molecular genetic characteristics, and differential diagnosis of NTRK-rearranged spindle cell neoplasms (NTRK-RSCNs) in the gastrointestinal tract. Methods: Two NTRK-RSCNs diagnosed at the Department of Pathology of the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China and one case diagnosed at Zhengzhou Central Hospital, Zhengzhou, China from 2019 to 2022 were collected. The clinical data, histopathology, immunophenotypes and prognosis were analyzed. Fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS) were used to detect NTRK gene rearrangements, while relevant literature was also reviewed and discussed. Results: Two patients were male and one was female, with the age of 17, 47 and 62 years, respectively. The tumors were located in the duodenum, ascending colon and descending colon, respectively. The tumors were protuberant masses with gray and rubbery sections. Their maximum diameter was 2.5, 5.0 and 10.0 cm, respectively. Histologically, the tumors invaded mucosa, intrinsic muscle and serosal adipose tissue. Tumor cells consisted of spindle or oval shaped cells with monotonous morphology and arranged in bundles or stripes pattern. Spindle cells were mildly to moderately atypical, with slightly eosinophilic cytoplasm and inconspicuous nucleoli. Necrosis and mitotic figures were observed in one high-grade tumor. All tumors expressed CD34, S-100 and pan-TRK in varying degrees. FISH analysis showed that NTRK1 gene was break-apart in 1 case and NTRK2 gene break-apart in 2 cases. NGS technologies showed LMNA::NTRK1 fusion in one case, STRN::NTRK2 fusion in another case. All patients recovered well after the surgery without recurrence at the end of the follow-up. Conclusions: NTRK-RSCN is rarely diagnosed in the gastrointestinal tract and has significant variations in morphology. It overlaps with various other mesenchymal tumors which should be considered as differential diagnoses. Be familiar with the features of histological morphology in combination with immunophenotype and molecular genetic characteristics can not only help diagnose NTRK-RSCNs, but provide therapeutic targets for clinical treatment.

The feasibility of traditional noninvasive prenatal diagnosis (NIPD) relying on proband-based relative haplotype dose analysis has been demonstrated. However, the prognosis of type I spinal muscular atrophy (SMA) is poor, and the proband sample is hard to collect during the second pregnancy. We investigate the feasibility of NIPD for SMA via haplotype construction without the need for a proband. Six samples were collected from both the paternal and maternal families in 36 families at risk of SMA. By enriching the SMN1/2 gene and its upstream and downstream informative SNPs, the family haplotype was constructed, and the Bayes factor was used to infer the fetal genotype by the dose changes of informational SNPs in cell-free DNA. All samples underwent MLPA testing after chorion villus sampling or amniocentesis. The MLPA results showed 100% consistency with NIPD. The earliest gestational week for successful NIPD was 7+ 3 weeks, with a minimum fetal fraction of 1.9%. Haplotype construction based on both paternal and maternal families demonstrated significant reliability and feasibility for families without a proband. Additionally, this approach provides a safer, and earlier prenatal diagnosis option for couples identified as at-risk through SMA carrier screening.

BackgroundFacioscapulohumeral muscular dystrophy 1 (FSHD1) is an autosomal dominant muscular disorder mainly caused by the contraction and hypomethylation of the D4Z4 repeat array in chromosome 4q35. Prenatal diagnosis of FSHD1 is challenging due to the highly repetitive and long genomic structure. In this study, a pregnant woman diagnosed with FSHD1 using optical genome mapping sought assistance for a healthy offspring.MethodsAt the 17th week of gestation, she underwent amniocentesis, and genomic DNA (gDNA) was extracted from amniocytes. Whole-genome sequencing of the gDNA was performed using the nanopore MinION platform.ResultsDespite a sequencing depth of only 7.3×, bioinformatic analyses revealed that the fetus inherited four D4Z4 repeat units with the permissive 4qA from the mother and the eight D4Z4 repeat units with the non-permissive 4qB from the father. To validate the results, SNP-based linkage analyses were conducted with gDNA from the proband, the proband’s father and proband’s amniocytes. Results indicated that the fetus inherited the maternal pathogenic haplotype based on 144 informative SNPs. Linkage analysis was consistent with the nanopore sequencing.ConclusionNanopore sequencing proves to be an accurate and direct method for genetic testing of monogenic diseases at the single-nucleotide level. This study represents the first application of nanopore sequencing in the prenatal diagnosis of FSHD1, providing a significant advantage for patients with de novo mutations.

ABSTRACT Background The clinical performance of RHDO‐based NIPD for PKU during early gestation remains under‐evaluated. Furthermore, studies focused on SNP loci obtained by next‐generation sequencing to analyze the genetic evolution of pathogenic variations in PKU is limited. Methods Maternal peripheral blood, along with proband and paternal samples, was collected between 7 and 12 weeks of gestation. The PAH gene and surrounding high heterozygosity SNPs were targeted for enrichment and sequencing. Fetal genotypes were inferred using RHDO‐based NIPD. High‐resolution PAH haplotypes were used for the analysis of two common pathogenic variants in the Chinese population: c.728G>A and c.1238G>C. Results Sixty one PKU families participated with an average fetal fraction of 6.08%. The median gestational age was 8 +6 weeks. RHDO‐based NIPD successfully identified fetal genotypes in 59 cases (96.72%, 59/62). Two cases failed because of insufficient informative SNPs. In addition, a recombination event was assessed in one fetus of 59 cases. Six, and three haplotypes were identified for c.728G>A(p.Arg243Gln) and c.1238G>C(p.Arg413Pro), respectively. Hap_3 and hap_8 were identified as the ancestral haplotypes for these pathogenic variants, with other haplotypes arising from mutations or recombination based on these ancestral haplotypes. Conclusions This study validates the feasibility of an RHDO‐based assay for NIPD of PKU in early pregnancy and introduces its application in the demonstration of founder effects in recurrent pathogenic variations, offering new insights into the evolutionary analysis of PAH variations.

ObjectiveTo establish a haplotype-based noninvasive prenatal testing (NIPT) workflow for single-gene recessive disorders that adapt to dizygotic (DZ) twin pregnancies.MethodTwin pregnancies at risk of Duchenne muscular dystrophy, Becker muscular dystrophy, hemophilia B, spinal muscular atrophy, phenylketonuria, and nonsyndromic hearing loss were recruited. For subsequent analysis, capture sequencing targeting highly heterozygotic single nucleotide polymorphism sites was conducted. Paternal-specific alleles were used to calculate the total and individual fetal fractions and determine zygosity. A two-step Bayes Factor model was applied to clarify the complex genomic landscape in the maternal plasma: the first step involved determining whether the twins inherited the same haplotype, and the second step involved estimating their individual genotypes. NIPT results were subsequently confirmed by invasive diagnosis.ResultsNine twin pregnancies were recruited, including five DZ and four monozygotic (MZ) twins. The earliest gestational age was 8+0 weeks, and the minimum fetal fraction was 4.6%. Three twin pregnancies were reported with one affected fetus, while the remaining six were reported without affected fetuses. Two dichorionic diamniotic twin pregnancies were confirmed to be MZ twins. The NIPT results were 100% consistent with those of invasive procedures or diagnostic genetic testing after birth.ConclusionThis study is the first to perform NIPT for single-gene disorders in twin pregnancies and preliminarily confirm its clinical feasibility. Acknowledging the twins' genotypes in the first trimester is valuable as it empowers obstetric care providers and parents to have adequate time for pregnancy management and decision-making. What's already known about this topic?The haplotype-based approach has been proven accurate for the noninvasive prenatal diagnosis of various autosomal or X-linked recessive single-gene disorders in singleton pregnancies. However, its applicability in dizygotic (DZ) twin pregnancies remains unclear.What does this study add?Targeted sequencing of maternal cell-free DNA followed by a two-step Bayes Factor model was used to evaluate fetal genotypes from nine twin pregnancies with 100% accuracy. Our study proved the applicability of haplotype-based noninvasive prenatal testing (NIPT) in DZ twin pregnancies. It further highlighted the importance of early pregnancy NIPT for the management and decision-making of twin pregnancies.

The intractability and high mortality rate of castration-resistant prostate cancer (CRPC) remain the most challenging problems in the field of prostate cancer (PCa). Emerging evidence has shown that the dysregulation of Wnt signaling pathways, which are highly conserved cascades that regulate embryonic development and maintain tissue homeostasis, is involved in various stages of PCa occurrence and progression. In this review, we systemically discuss the mechanisms by which the androgen receptor (AR) signaling pathway and Wnt signaling pathways participate in the occurrence of PCa and its progression to CRPC. Specifically, we elaborate on how Wnt signaling pathways induce the malignant transformation of prostate cells, promote the malignant progression of PCa and establish an immunosuppressive prostate tumor microenvironment through interaction with the AR pathway or in an AR-independent manner. We also discuss how Wnt signaling pathways enhances the stemness characteristics of prostate cancer stem cells (PCSCs) to induce the occurrence and metastasis of CPPC. Additionally, we discuss the latest progress in the use of different types of drugs that inhibit the Wnt signaling pathways in the treatment of PCa. We believe that the combination of Wnt signaling-based drugs with endocrine and other therapies is necessary and may enhance the clinical efficacy in the treatment of all types of PCa.