OBJECTIVE: To explore the clinical characteristics and genetic etiology of STISS syndrome (an autosomal dominant disorder characterized by ubiquitin-proteasome system dysfunction) in a child. METHODS: A child with STISS syndrome diagnosed at the Affiliated Women and Children's Hospital of Ningbo University in September 2024 due to "abnormal development of external genitalia" was selected as the study subject. Clinical data were retrospectively collected. Peripheral blood samples were obtained from the child and his family members. Following genomic DNA extraction, whole-exome sequencing (WES) and Sanger sequencing validation were carried out. Pathogenicity of the candidate variants was assessed based on the guidelines from American College of Medical Genetics and Genomics (ACMG). The study was approved by the Ethics Committee of the hospital (Ethics No.: EC2023-094). RESULTS: The proband, a 16-year-old boy, presented with micropenis, testicular hypoplasia, delayed sexual development, insulin resistance, diabetes mellitus, and obesity. WES revealed that he has harbored a c.934del; p.Met312TrpfsTer3 frameshifting variant of the PSMD12 gene, which was unreported previously. Sanger sequencing confirmed that the variant to be de novo in origin. Based on the guidelines from the ACMG, the variant was classified as pathogenic (PVS1+PM2_supporting+PM6_supporting). The variant was predicted to result in a premature termination codon. Bioinformatics analysis suggested that the amino acid at position 312 is highly conserved, and the variant may therefore affect the protein structure. CONCLUSION Patients with STISS syndrome exhibit clinical features including psychomotor retardation, intellectual disability, distinctive facial features, and urogenital abnormalities. The c.934del (p.Met312TrpfsTer3) frameshift variant of the PSMD12 gene probably underlay the pathogenesis in the proband. Above finding has enriched the mutational spectrum of the PSMD12 gene.
Adolescent ; Humans ; Male ; Exome Sequencing ; Mutation ; Proteasome Endopeptidase Complex/genetics* ; Syndrome

OBJECTIVE: To optimize DNA library construction in non-crosslinked chromatin immunoprecipitation coupled with next-generation sequencing (Native ChIP-seq) to obtain high-quality Native ChIP-seq data. METHODS: Human nasopharyngeal carcinoma HONE1 cell lysate was digested with MNase for release of the nucleosomes, and the histone-DNA complexes were immunoprecipitated with specific antibodies. The protein component in the precipitate was digested with proteinase K followed by DNA purification; the DNA library was constructed for sequence analysis. RESULTS: Compared with the conventional DNA library construction, Tn5 transposase method allowed direct enrichment of the target DNA after Tn5 fragmentation, which was simple, time-saving and more efficient. The IGV visualized map showed that the information obtained by the two library construction methods was consistent. The sequencing data obtained by the two methods revealed more signal enrichment with Tn5 transposase library construction than with the conventional approach. H3K4me3 ChIP results showed a good reproducibility after Tn5 transposase library construction with a signal-to-noise ratio above 50%. CONCLUSIONS Tn5 transposase method improves the efficiency of DNA library construction and the results of subsequent sequence analysis, and is especially suitable for detecting histone modification in the DNA to provide a better technical option for epigenetic studies.
Chromatin Immunoprecipitation ; DNA ; Gene Library ; High-Throughput Nucleotide Sequencing ; Humans ; Reproducibility of Results ; Sequence Analysis, DNA

BACKGROUND: Total hip arthroplasty is an effective measure to treat hip involvement in ankylosing spondylitis.Ankylosing spondylitis patients have different degrees of anemia after total hip arthroplasty. The hidden blood loss accounts for a large proportion of perioperative blood loss in total hip arthroplasty, and can affect the recovery of joint function.OBJECTIVE: To investigate risk factors of hidden blood loss after total hip arthroplasty in patients with hip involvement in ankylosing spondylitis.METHODS: We studied a consecutive series of 70 hips in 60 patients with ankylosing spondylitis hip involvement who were converted to cementless total hip arthroplasty. The average age of surgery was 35.12 years. The hidden blood loss was calculated according to Cross formula linear equation. The effects of operation time, erythrocyte sedimentation rate,C-reactive protein, body mass index, Bath ankylosing spondylitis radiology index, allogenic blood transfusion, and osteoporosis on hidden blood loss after total hip arthroplasty in patients with ankylosing spondylitis were analyzed. The patients were divided into the high blood loss group (≥ 480 mL) and the low blood loss group (< 480 mL) according to the high blood loss. Risk factors of high hidden blood loss after total hip arthroplasty in patients with ankylosing spondylitis were analyzed by single factor analysis and multivariate Logistic regression analysis (SPSS 17.0).RESULTS AND CONCLUSION: (1) The hidden blood loss after primary total hip arthroplasty in patients with ankylosing spondylitis was (737.76±419.18) mL, and the total blood loss was (1312.83±487.41) mL, and the percentage of hidden blood loss was 51.48%. The high blood loss group included 41 hips, and the low blood loss group included 29 hips; and the ratio was 41:29. (2) Single factor analysis showed that the operation time, Bath ankylosing spondylitis radiology index and osteoporosis, allogenic blood transfusion, decrease of hemoglobin were significantly associated with high hidden blood loss. (3) Multivariate Logistic regression analysis showed that Bath ankylosing spondylitis radiology index,allogeneic blood transfusion, and decrease of hemoglobin were significantly associated with high hidden blood loss. (4)Hidden blood loss is an important portion of total blood loss after primary total hip arthroplasty in patients with ankylosing spondylitis. Bath ankylosing spondylitis radiology index, allogeneic blood transfusion and decrease of hemoglobin are risk factors for high hidden blood loss.