Objective: To explore the effectiveness and safety of the ketogenic diet (KD) in children with drug resistant epilepsy (DRE) caused by structural etiology.Methods: The children were categorized into acquired brain injury group and malformations of cortical development (MCD) group based on the etiology. Follow-up assessments were performed at 1, 3, and 6 months after KD treatment to observe seizure reduction, behavioral and cognitive improvements, adverse reactions events, and reasons for discontinuation withdrawal. Statistical analysis was conducted on the results.Results: We found the seizure-free rates at 1, 3, and 6 months were 4.8% (2/42), 19% (8/42), and 21.4% (9/42), respectively. The seizure control effective rates were 42.9% (18/42), 52.4% (22/42), and 54.8% (23/42) at the corresponding time points. Compared to the acquired brain injury group, the MCD group showed a higher seizure control effective rate. Further analysis within the MCD group revealed the highest efficacy in focal cortical dysplasia (FCD). At the 3-month follow-up, cognitive and behavioral improvements were observed in 69% (29/42) of children. The main reasons for discontinuation were lack of efficacy and poor compliance.Significance: Finally, we get that KD is a safe and effective treatment for drug resistant epilepsy caused by structural etiology, with the added benefit of improving behavioral and cognitive abilities in children. The efficacy is higher in children with MCD, particularly in cases of FCD. Early intervention with KD is recommended for this population.

Inherited retinal diseases (IRDs) constitute a complex and heterogeneous group of rare disorders characterized by significant genetic diversity. These conditions often lead to the degeneration of photoreceptor cells, resulting in severe visual impairment. A major challenge in the study and treatment of IRDs is the lack of appropriate preclinical models for investigating their pathogenesis and evaluating potential therapeutic interventions. In recent years, advances in retinal organoids (ROs) culture technology have provided promising new avenues for IRDs. This review systematically elaborates on the applications of induced pluripotent stem cells (iPSC)-derived ROs as disease models in IRDs such as retinoblastoma (RB), retinitis pigmentosa (RP), Leber congenital amaurosis (LCA), and X-linked juvenile retinoschisis (XLRS). This review also briefly explores the culturing methods of iPSC-derived ROs in recent years. Specifically, it emphasizes the comparison of the similarities between the formation process of ROs and the in-vivo retinal development process. This encompasses aspects such as the formation of different structures, the expression of key markers, and the manifestation of physiological functions. By utilizing the RO disease model to explore the pathogenesis of IRDs and conduct drug screening, it is expected to advance the precision treatment of IRDs and improve the therapeutic outcomes for patients.

FLNC gene variants have predominantly been reported in adult populations with cardiomyopathies, and early-onset cases are less common. The genotype–phenotype relationship indicates that dilated cardiomyopathy (DCM) is often associated with FLNC truncating variants. We conducted a comprehensive genetic analysis using next generation sequencing (NGS) to identify FLNC variants in patients with cardiovascular conditions. Detailed phenotypic and variant analyses were performed to characterize the clinical features and genetic alterations. Minigene assays and structural modeling were used to investigate the pathogenicity caused by the identified variants. In a cohort of 58 patients, novel heterozygous FLNC variants, c.3962A > T (p.Glu1321Val) and c.7543C > T (p.Leu2515Phe), were identified in patients presenting with dilated and mixed restrictive/hypertrophic cardiomyopathies, respectively. The c.3962A > T variant disrupted normal splicing, as demonstrated through the splicing prediction tool and minigene studies, further emphasizing its pathogenic potential. For missense variants of FLNC in patients with DCM, the splicing effect of the variant should be carefully checked. Early detection and intervention are crucial given the high risk of sudden cardiac death and severe cardiac complications.

Gestational diabetes mellitus (GDM) is a common metabolic disorder during pregnancy, involving multiple immune and inflammatory factors. Macrophages play a crucial role in its development. This study integrated scRNA-seq and RNA-seq data to explore macrophage-related diagnostic genes and GDM subtypes. For scRNA-seq data, cell clusters were annotated using the SingleR package and validated with marker gene expression profiles, while hdWGCNA analysis identified three gene modules related to macrophages. A diagnostic model for GDM derived from endothelial cell transcriptomes was constructed by employing a variety of machine learning ensemble algorithms, achieving an AUC of 0.887. The model identified five differentially expressed genes (ZEB2, MALAT1, HEBP1, AHSA1, and TTC3) as potential diagnostic biomarkers. The CB-DSNMF algorithm was proposed to identify two distinct GDM subtypes from RNA-seq data, revealing significant differences in biological behaviours. This algorithm outperformed other baselines in multiple clustering metrics. Mendelian randomisation analysis identified ZEB2 as a gene causally related to GDM risk. A transcription factor (TF)-gene regulatory network was constructed for these genes using the ENCODE database. The study highlights the importance of macrophages in GDM, provides a high-precision diagnostic model, and offers new insights into personalised treatment strategies, contributing to a better understanding of GDM pathophysiology.

Sitosterolemia is classically described as an autosomal recessive disorder caused by biallelic pathogenic variants in ABCG5 or ABCG8. Emerging evidence suggests that heterozygous carriers may exhibit subtle biochemical or clinical features, prompting a re-evaluation of its inheritance paradigm. This study aimed to characterize the clinical and biochemical phenotypes—particularly childhood linear growth and plasma phytosterol levels—in individuals harboring ABCG5 variants, and to explore the association between specific genotypes and phytosterol accumulation. Clinical and genetic data from three families with ABCG5 variants were retrospectively analyzed, and childhood height was evaluated relative to mid-parental target height. In one family with unexplained markedly elevated sitosterol, whole-genome sequencing (WGS) was performed to investigate potential noncoding variants. A candidate variant, c.-162 T > C (p.( =)), was annotated using the Ensembl Variant Effect Predictor (VEP) and functionally evaluated by dual-luciferase assays. Participants were stratified by genotype into wild-type (WT), heterozygous (Het), and compound heterozygous (cHet) groups to compare plasma sitosterol levels. Four children with ABCG5 variants had elevated plasma sitosterol and heights below mid-parental targets, with their heights all falling between the 3rd and 20th percentile; individuals with biallelic ABCG5 variant combinations showed more pronounced growth impairment. Their parents all had normal adult heights. Sitosterol levels varied by genotype: heterozygous c.1336C > T (p.Arg446Ter) (41.7 μmol/L); a compound heterozygote with c.1336C > T (p.Arg446Ter) and ABCG5 exons 1–4 deletion (668.4 μmol/L); a trans-heterozygote carrying c.1336C > T (p.Arg446Ter) and c.-162 T > C (p.( =)) (193.9 μmol/L); and a heterozygous carrier of c.-162 T > C (p.( =)) showing mildly elevated sitosterol (19.2 μmol/L). The c.-162 T > C (p.( =)) variant is a 5′ UTR variant in the second most abundant ABCG5 liver isoform; dual-luciferase assays showed a modest, non-significant reduction in promoter activity, but combined clinical, genetic, and biochemical data support its potential pathogenicity. Sitosterol levels exhibited a trend consistent with gene dosage, with the highest concentrations in the cHet group, intermediate levels in the Het group, and the lowest in the WT group. Biallelic ABCG5 variants are associated with childhood growth impairment. The c.-162 T > C (p.( =)) variant is likely pathogenic, though its role requires further validation. Integrated sterol profiling and comprehensive genetic analysis in familial contexts are essential for clarifying clinical status and inheritance.

Maple syrup urine disease (MSUD) is an inherited metabolic disorder caused by a deficiency in the activity of the hepatic branched-chain α-ketoacid dehydrogenase (BCKDH) complex, which leads to the toxic accumulation of three branched-chain amino acids (BCAAs) and their respective α-ketoacid, resulting in severe neurotoxicity, coma and even death without effective therapeutic measures. In this study, we established the patient induced pluripotent stem cells (iPSC)-derived hepatic organoids (HOs), analyzed the characteristics, and applied adenine base editor (ABE8e) to correct a mutation (T322I) of the BCKDHB (branched chain keto acid dehydrogenase E1, beta polypeptide) gene in patient induced pluripotent stem cells (iPSC)-derived hepatic organoids (HOs). qRT-PCR and western blot analysis were performed to assess the expression level of BCKDHA (branched chain keto acid dehydrogenase E1, alpha polypeptide) and BCKDHB. The effects of base editing were comprehensively analyzed using both bulk RNA sequencing and single-cell RNA sequencing (scRNA-Seq). Immunofluorescence and RT-PCR arrayed the high expression of hepatoblast specific proteins in HOs, such as α-1-anti-trypsin (A1AT), hepatocyte nuclear factor-4-alpha (HNF4A), cytokeratin18 (CK18), albumin (ALB), cytochrome P450 family 3 subfamily A member 4 (CYP3A4) and cytochrome P450 family 3 subfamily A member 7(CYP3A7). Functional experiments indicated that these HOs recapitulated characteristics of hepatocytes like glycogen accumulation, low-density lipoprotein (LDL) uptake, indocyanine green (ICG) uptake and release as well as quantitation of ALB and urea from HOs. The levels of BCKDHA and BCKDHB were dramatically decreased in MSUD-HOs compared with control-HOs (P < 0.01) detected by qRT-PCR, western blot and immunofluorescence. Deep sequencing and whole genome sequencing (WGS) demonstrated that the correction of BCKDHB mutation in patient iPSC-derived HOs caused high on-target gene editing without any detectable off-target effects. Moreover, the corrected MSUD-HOs exhibited restored BCKDH enzymatic function and reduced BCAAs level. The transcriptome analysis indicates that the MSUD-HOs with BCKDHB mutation reduced mRNA level of regulating metabolism associated with liver mitochondrial function, while the corrected MSUD-HOs rescued those processes after ABE8e correction. The scRNA-Seq analysis further validated the rescue effects of BCKDH function after gene editing. Our study provides reliable evidence that ABE8e is highly efficient and safe in correcting patient-derived HOs from MSUD, indicating the feasibility to be a transformative treatment for genetic hepatic diseases like MSUD.