OBJECTIVE: To apply optical genome mapping (OGM) technique for the analysis of genetic etiology in two rare families with complex chromosomal rearrangements (CCRs) and to provide precise reproductive guidance to them. METHODS: Two Chinese families diagnosed with chromosomal rearrangements by chromosomal microarray analysis (CMA) or whole-exome sequencing (WES) between June and December 2023 at the Affiliated Women and Children's Hospital of Ningbo University were selected as the study subjects. In both cases, unbalanced chromosomal translocations were suspected. Clinical data were collected, and peripheral blood from the couple, amniotic fluid sample and aborted fetal tissue was subjected to combined G-banding karyotyping and OGM for comprehensive genetic analysis. This study has been approved by the Medical Ethics Committee of the Hospital (Ethics No.: EC2023-094). RESULTS: In family 1, the fetus was signaled to have abnormal chromosome 7 by non-invasive prenatal testing (NIPT), prompting amniocentesis and CMA detection. In family 2, a pregnancy loss had occurred at 10 weeks' gestation, and trio-WES was carried out. Both fetuses were found to harbor copy number variations (CNVs) suggestive of unbalanced CCRs. Further analysis with OGM has revealed that, in family 1, an unbalanced rearrangement involving chromosomes 7, 8, and 10 was carried by the fetus and the pregnant woman, which has formed der(8) and der(10) derivative chromosomes. In family 2, a maternal CCR was found, which involved chromosomes 2 and 13 with seven breakpoints, resulting in unbalanced fetal CNVs. After genetic counseling, family 1 opted to continue with the pregnancy, considering the woman's normal appearance and inheritance of the rearrangement. For both families remained to have a risk for unbalanced rearrangements in subsequent pregnancies, preimplantation genetic testing (PGT) was recommended. CONCLUSION In both families, the OGM has precisely delineated the genetic basis of fetal CNVs and mapped the maternal CCR breakpoints, providing critical insights for genetic counseling and reproductive decision-making.
Adult ; Female ; Humans ; Male ; Pregnancy ; Chromosome Aberrations ; Chromosome Disorders/genetics* ; Chromosome Mapping/methods* ; Genetic Testing/methods* ; Pedigree ; Prenatal Diagnosis/methods* ; Translocation, Genetic

OBJECTIVE: To assess the value of Optical genome mapping (OGM) for the verification of chromosomal structural variations among patients undergoing assisting reproduction. METHODS: A retrospective analysis was carried out on the clinical data of 12 patients presented at the Reproductive Center of Ningbo University Women and Children's Hospital from October 2022 to October 2024. All patients had undergone OGM testing due to suspection of structural variants by chromosomal karyotyping or a suggestive medical history. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: EC2024-148). RESULTS: Among the 12 patients verified by OGM, one (8.3%) was in keeping with the result of chromosomal karyotyping. Revised karyotypes were confirmed in seven cases (58.3%), including four with complex chromosomal rearrangements. Structural variation was excluded in three cases (25.0%). Of note, OGM has identified a previously undetected cryptic balanced translocation, i.e., ogm[GRCh38] t(7;12)(q36.3;q24.23)(157511190_157523142;119205703_119198409). CONCLUSION OGM can serve as an auxiliary diagnostic technique to conventional karyotyping and enable validation of suspected structural variations in those with ambiguous karyotype results or a history of adverse pregnancies. This can provide more precise genetic diagnosis for patients undergoing assisted reproduction and selection of clinical intervention strategies.
Humans ; Female ; Adult ; Retrospective Studies ; Karyotyping ; Reproductive Techniques, Assisted ; Chromosome Mapping/methods* ; Chromosome Aberrations

OBJECTIVE: To investigate the genetic characteristics and clinical utility of Optical genome mapping (OGM) in resolving complex genomic rearrangements in families with recurrent pregnancy loss. METHODS: A recurrent miscarriage family which presented at both the People's Hospital of Qianxinan Buyi and Miao Autonomous Prefecture and the Affiliated Women and Children's Hospital of Ningbo University in September 2024 was selected as the study subject. Relevant clinical information was collected. Peripheral blood samples of the couple were collected for G banding karyotyping analysis, and copy number variation sequencing (CNV-seq) and OGM were used for verification. This study was approved by the Medical Ethics Committee of the Affiliated Women and Children's Hospital of Ningbo University (Ethics No.: EC2024-148). RESULTS: CNV-seq in an external hospital detected a 10.67 Mb deletion in the 16q12.1q21 region, a 142.4 kb deletion in the 5p15.2 region, and a 359.55 kb duplication in the 7p22.2 region. No abnormality was found in the chromosomal karyotype of the male partner, and the initial karyotyping of the female partner suggested 46,XX,?del(16)(q12.1q22). The CNV-seq verification of her indicated only variations in the 5p15.2 and 7p22.2 fragments, and no deletion of 16q was detected. As indicated by precise OGM analysis, multiple intrachromosomal and interchromosomal translocation variations had occurred between chromosomes 10 and 16 in the female partner, with complex balanced rearrangements (including 5 transchromosomal breakpoints). CONCLUSION The complex balanced rearrangements of the female partner's chromosomes had occurred during meiosis, the resultant unbalanced gametes may be the cause of repeated miscarriage in this family. OGM can delineate complex rearrangement breakpoints and directions that are difficult to reveal by conventional karyotyping analysis and provide a basis for accurate reproductive genetic counseling.
Humans ; Abortion, Habitual/etiology* ; Female ; Pregnancy ; Male ; DNA Copy Number Variations/genetics* ; Adult ; Karyotyping ; Pedigree ; Gene Rearrangement ; Chromosome Mapping

Resting-state functional MRI(rs-fMRI)is an important tool for non-invasive research of brain function having been widely used in gray matter network function but not in changes of white matter brain function in neurological and psychiatric disorders.Recent studies on white matter function in neuropsychiatric disorders had provided new insights into the mechanisms underlying brain network organization and pathogenesis of brain disorders.The progresses of rs-fMRI research on brain white matter networks in neuropsychiatric diseases were reviewed in this article.

Objective To analyze the modeling and evaluation method for vascular restenosis animal models in the last 10 years,to provide a reference for improving animal models of vascular restenosis.Methods Literature related to vascular restenosis was retrieved from mainstream Chinese and English databases from 2013 to 2023.Data on experimental animal strains,modeling method,modeling cycles,and detection method were extracted from the included literature,and a database was established using Excel for summary analysis.Results Among the 122 identified articles,the main experimental animals were rats,rabbits,and pigs,and most animals were male.The most common modeling method was balloon injury,and the modeling cycle was mainly within 4～8 weeks.The main detection indexes were histopathology,accounting for 37.18％,including routine hematoxylin-eosin,Masson,and Elastica van Gieson(EVG)staining.Conclusions The translatability of porcine vascular restenosis models is currently more in line with expectations,but their cost is high and they are unpopular,and rats and rabbits thus remain the main animal models.Balloon injury is the main mode of modeling.Different animal models and modeling method for vascular restenosis have advantages and disadvantages,and the model should be selected according to the experimental purpose.Animal models of vascular restenosis still have some limitations,however,and better animal models are required in the future.

Peritoneal dialysis (PD) is a crucial renal replacement therapy for end-stage renal disease (ESRD), offering significant advantages as high flexibility, hemodynamic stability, and high cost-effectiveness. However, prolonged exposure to intra-abdominal dialysate may predispose to the mechanical complication of abdominal external hernia. Abdominal external hernia may lead to various adverse clinical outcomes. In severe cases, it can progress to incarceration or even rupture, ultimately necessitating discontinuation of the therapy. The authors systematically review PD-associated abdominal external hernias, including their clinical landscape, risk factors, surgical treatment strategies and prognostic determinants. They also assess the effects of hernia repair on residual renal function, aiming to provide references for clinical decision-making.

Objective To explore the application value of metagenomic next-generation sequen-cing(mNGS)technology in the diagnosis of spinal tuberculosis.Methods A total of 129 patients with suspected spinal tuberculosis admitted from January 2021 to January 2023 were selected as study subjects.Lesion tissue samples were collected intraoperatively and subjected to conventional microbio-logical testing(CMT),Mycobacterium tuberculosis DNA(MTB-DNA)amplification testing,and mNGS testing.The diagnostic efficacy of different testing methods was compared using results of com-prehensive clinical diagnosis as the gold standard.Results Among 129 patients,101(78.29％)were confirmed to have spinal tuberculosis,and 28(21.71％)had other spinal infections.Using clinical results as the diagnostic gold standard,the sensitivity of mNGS was 94.06％(95/101),and specificity was 89.29％(25/28);the sensitivity of MTB-DNA amplification was 90.10％(91/101),and specificity was 89.29％(25/28);the sensitivity of CMT was 86.14％(87/101),and specifici-ty was 85.71％(24/28).Compared with MTB-DNA amplification and CMT,mNGS showed the highest consistency with clinical results,and its consistency in detecting different lesion sites was also optimal,with statistically significant differences(P＜0.05).Conclusion mNGS testing has high diagnostic value for spinal tuberculosis and can provide a reference for clinical diagnosis,thereby guiding clinical decision-making.

Objective: To study the effect of metformin sensitizing pancreatic cancer cells with radiotherapy, with a focus on elucidating the underlying mechanisms of radiotherapy resistance. In particular, the role of the PERK/P-eIF2/ATF4 signaling pathway in mediating these effects was preliminarily explored. Methods : Pancreatic cancer cell lines(PANC-1 and PANC-2) were categorized into control, radiotherapy, and drug treatment groups. Following the respective treatments, cell proliferation inhibition was assessed using the CCK-8 assay, colony formation assays, and cell death staining. Senescence was quantified by β-galactosidase(SA-β-Gal) staining. The expression of cell cycle regulators(P21, P16, γ-H2AX), apoptosis markers(Bax, Bcl-2, Cleaved caspase-3), and pathway-related proteins(PERK, P-eIF2, ATF4) was evaluated by Western blot and immunofluorescence. To further investigate the role of the PERK/P-eIF2/ATF4 axis in metformin-mediated modulation of pancreatic cancer cell senescence and radiosensitization, selective inhibitors(GSK2606414) and agonists(MK-28) of PERK were employed. Results : Radiotherapy markedly upregulated senescence-associated markers(P21, P16, γ-H2AX, and β-galactosidase activity) in pancreatic cancer cells. Senescent cells exhibited enhanced proliferative activity and increased tumor volume both in vitro and in vivo. Metformin mitigated radiotherapy-induced senescence by reducing the expression of senescence markers and significantly suppressing the clonogenic and proliferative capacity of treated cells. Mechanistically, radiotherapy activated the PERK signaling pathway, leading to increased expression of PERK, P-eIF2, and ATF4, thereby driving cellular senescence. Pharmacological inhibition of PERK reduced β-galactosidase activity, while PERK activation further promoted the expression of senescence-associated proteins—an effect that was reversed by metformin. Conclusion Metformin inhibits the activation of the PERK/P-eIF2/ATF4 signaling pathway in pancreatic cancer cells following radiotherapy, thereby delaying cellular senescence and reducing the associated radiotherapy resistance of senescent cells. This modulation contributes to the sensitization of pancreatic cancer cells to radiotherapy.

Objective:To develop a method for determining 14C in seafood using a tube combustion furnace (or oxygen bomb combustion devices) coupled with liquid scintillation counting (LSC), in order to accurately determine 14C in seafood. Methods:Four categories of seafood samples (i.e., fish, crustaceans, mollusks, and algae) were collected. They were then subjected to high-temperature oxidation using a tube combustion furnace or oxygen bomb combustion devices to isolate CO 2, followed by an analysis of the radioactivity concentration of 14C using LSC. The combustion conditions were optimized by investigating the heating nodes and rates in the oxidation combustion furnace, and the CO 2 collection conditions were optimized by placing a NaOH absorption solution in the oxygen bomb combustion devices. Additionally, the optimal measurement conditions were determined by comparing the effects of varying scintillation cocktails and the dark adaptation time of two preparation methods, i. e., the adsorption and suspension method. Results:When a tubular combustion furnace was used for oxidation and combustion, the optimal heating rate of the sample pyrolysis temperature zone was determined at 1.5℃/min. In this case, the combustion efficiency of various seafood could reach over 95%. When the oxygen bomb combustion devices were employed, placing NaOH solutions both inside and at the exhaust end increased absorption efficiency by 10% compared to the traditional practice of placing a NaOH solution only at the exhaust end. Samples prepared using the absorption method should be kept in the dark for at least 10 h before measurement using LSC, while samples prepared with the suspension methods should be kept for at least 20 h. The results obtained using two preprocessing devices and two sample preparation method were consistent, with a detection limit of 9.10 Bq/kg (dry samples). Compared to the suspension method (The relative standard deviation of the results obtained by two preprocessing devices were 9.00% and 8.27%), the absorption method (the corresponding relative standard deviation were 3.61% and 3.29%) exhibited higher precision in repeated measurements.Conclusions:The aforementioned pretreatment devices and sample preparation method are suitable for determining 14C in seafood.

Energy metabolism disorders are a critical factor contributing to cognitive dysfunction.Astrocytes,as key suppliers of energy substrates to neurons,help to sustain normal cognitive function via their extensive energy metabolism activities in the brain,coordinated by various energy supply mechanisms.Imbalances in astrocyte-neuron energy metabolism coupling lead to abnormal neuronal activity,thereby driving the pathogenesis and progression of Alzheimer's disease(AD).It is therefore essential to investigate the pathological mechanisms and therapeutic targets associated with AD from the perspective of energy metabolism to support the future development of AD treatments and interventions.Traditional Chinese medicine(TCM),characterized by multi-component and multi-target effects,has shown potential efficacy for regulating cerebral energy metabolism,suggesting that modulating brain energy metabolism may represent a significant pathway by which TCM might ameliorate AD.This review systematically elucidates the mechanistic role of the dysregulation of astrocyte-neuron bioenergetic coupling in the pathogenesis and progression of AD,and critically evaluates recent advancements in therapeutic interventions mediated by TCM.