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

Objective To analyze the gene expression of patients with head and neck squamous cell carcinoma(HNSCC)using bioinformatics methods,identifying immune-related genes that impact prognosis and exploring new therapeutic targets for HNSCC.Methods This study was based on The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus database to download RNA sequencing data from HNSCC and non-tumor tissue samples.Differential expression gene analysis and weighted gene co-expression network analysis were performed respectively to screen immune-related genes.Univariate Cox regression,Lasso regression and multifactor Cox regression analyses were used to identify the genes significantly associated with HNSCC prognosis,and on this basis,the risk score of each TCGA sample was calculated and the correlation prognosis model was constructed.The sample was divided into high and low risk groups according to the median risk score.Gene differential expression analysis was performed in high and low risk groups to reveal gene expression changes under different immune states,and pathway enrichment analysis was performed for differential genes.Results Screening identified MS4A1,IL12RB2,DMBT1 and LTF as immune-related genes affecting the prognosis of HNSCC.Among them,MS4A1 and IL12RB2 were highly expressed in HNSCC,and DMBT1 and LTF were lowly expressed.Risk score of HNSCC death group was significantly higher than that of the survival group.Risk score could be used as an independent prognostic indicator for HNSCC.Differential genes in high and low risk groups were significantly enriched in immune response-regulating cell surface receptor signaling pathways.Conclusion MS4A1,IL12RB2,DMBT1 and LTF can be used for risk assessment,disease surveillance and efficacy evaluation of HNSCC.The immune response is crucial in the prognosis of HNSCC patients,and further studies are expected to drive the development of new therapeutic strategies.

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 (Ethic 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.

Objective To explore the biomechanical safety of applying traditional Chinese orthopedic manipulation therapy after anterior cervical discectomy and fusion(ACDF)surgery,so as to provide a theoretical basis for clinical treatment in biomechanics.Methods Based on CT data,a three-dimensional finite element model of the normal C0-T1 cervical spine was established,and an ACDF postoperative finite element model of the C5-6 segment was constructed on this basis.Cervical spine rotation manipulation was simulated at the C4 and C7 segments of both models,and the von Mises stresses of the vertebral body,bilateral facet joints,intervertebral discs,and internal fixation system under manipulation loading of the C4 and C7 segments in both models were compared and analyzed.Results When the C4 segment was manipulated,the stress on the C5,C6,and C7 vertebral bodies in the ACDF postoperative model decreased by 12.3％,11.5％,and 26.4％,compared to the normal model.The stress on the left facet joints of the C4-5,C5-6,and C6-7 segments decreased by 12.3％,58.8％,and 15.4％,and the stress on the right facet joints decreased by 16.6％,92.1％,and 17.2％.The stress on the C4-5 and C6-7 segments decreased by 13.2％and 4.0％,while the maximum stress of the fusion cage,titanium plate,and screws in the C5-6 segment were 9.349,111.9,and 300.8 MPa.When the C7 segment was manipulated,the stress on the C4,C5,and C6 vertebral bodies in the ACDF postoperative model increased significantly compared to the normal model,especially the C5 vertebral body,with an increase of nearly 18 times.Except for the stress on the left facet joint of the C4-5 segment increased by 57.7％,the stress on the bilateral facet joints of other segments generally decreased,but the stress on the C4-5 and C6-7 segments increased by 43.2％and 21.7％and the stresses on the fusion cage,titanium plate,and screws in the C5-6 segment were 2.926,205.4,and 256.2 MPa.Conclusions The safety of performing manipulation on the upper vertebral body of the fusion segment after ACDF surgery is relatively high,but performing manipulation on the lower vertebral body of the fusion segment may lead to stress concentration and increase the risk of injury.When postoperative conservative treatment is implemented,the manipulation safety and indications should be considered to avoid operations in high-risk areas,and more precise and safe manipulation intervention treatment should be implemented based on the specific postoperative biomechanical state of the patient.

Diabetic cataract, a prevalent ocular complication of diabetes mellitus, arises from a complex interplay of pathological mechanisms, with oxidative stress and glycation stress playing central roles. Autophagy, a critical cellular self-protection mechanism, sustains intracellular homeostasis by selectively degrading damaged organelles and misfolded proteins, thereby counteracting the detrimental effects of oxidative and glycation stress under hyperglycemic conditions. Emerging evidence indicates a synergistic interaction between glycation stress and oxidative stress, which may exacerbate autophagic dysfunction and accelerate the onset and progression of diabetic cataract. However, the precise molecular mechanisms underlying this relationship remain incompletely understood. This review systematically examines the regulatory role of autophagy inthe pathogenesis of diabetic cataract, with a particular focus on how autophagic impairment influences disease progression under the combined effects of glycation and oxidative stress. By elucidating these mechanisms, the paper aims to provide novel insights into molecular diagnostic approaches and targeted therapeutic strategies for diabetic cataract.

Adult-onset Still's disease(AOSD)is a rare autoinflammatory disease characterized by fever,rash,arthritis,liver,spleen and lymph node enlargement,increased total number of peripheral white blood cells and neutrophil ratio.This paper reported a case of AOSD with dermal lymphadenitis(DL)complicated with hemophagocytic syndrome(HPS),in order to improve the clinicians'understanding for this complicated complication.The patient was a 48-year-old female who was admitted to the hospital with the complant of"intermittent fever with rash for 15 d".After 10 d of active anti-infection treatment,the symptoms were not improved,and there were new large congestive edematous erythema on the face and trunk,muscle pain in limbs and joints,and spleen enlargement.Laboratory tests showed increased white blood cell count,significantly decreased platelet count,hypofibrinogenemia,elevated serum ferritin,and elevated soluble interleukin-2 receptor sCD25;DL was pathologically diagnosed by axillary lymph node biopsy.After excluding other diseases,the diagnosis was confirmed as AOSD with DL complicated with HPS.After diagnosis of HPS,the patient was treated with hemophagocytic lymphohistiocytosis(HLH)-1994 regimen combined with rucotinib for 6 weeks,and the symptoms were improved;the patrent was discharged.The diagnosis of AOSD is particularly complex when complicated with the complications such as HPS,which requires carefully differential diagnosis,especially to exclude lymphoma.The cases of AOSD with DL are rare,and its etiology and pathogenesis need further study;early diagnosis and multidisciplinary collaboration are essential to improve the patient's prognosis.

Objective To explore the biomechanical characteristics of different types of lumbar disc herniation(LDH),and provide a theoretical basis for the clinical classification,diagnosis,and treatment of LDH.Methods A normal lumbar spine model with spinal nerves and four types of LDH models(central type,paracentral type/lateral recess type,intervertebral foramen type,extreme type)were constructed,and the biomechanical characteristics of the intervertebral discs and nerve roots in neutral posture and under six degrees of freedom conditions were analyzed.Results In neutral position,the stress of normal intervertebral disc was carger than that of the herniated intervertebral disc,and the stress ratio of annulus fibrosus and nucleus pulposus in normal model was greater than that of annulus fibrosus and nucleus pulposus in herniated models.In neutral position,the disc stress was mainly concentrated at the incisor between the protrusion and normal part,and recruitment occurred at outer edge of the disc.In flexion and extension position,the stress of the herniated intervertebral disc shifted to the right side.The maximum stress of herniated intervertebral disc appeared during right flexion,and the nerve root stress on the right side was carger than that on the left side.The stress of the herniated intervertebral disc during right rotation was slightly lower than that during right flexion,and the stress of nerve root on the right side was smaller than that on the left side.Conclusions The intervertebral disc is important in maintaining lumbar dynamic and static stability and flexibility.Different positions have an obvious effect the intervertebral disc stress.For patients with LDH,forward bending,flexion of the affected side and large extension should be avoided.Appropriate rotation of the affected side can help relieve the symptoms of nerve root compression.

Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.