Peking University Third Hospital

Growing evidence suggests that abnormal lipid metabolism occurs in amyotrophic lateral sclerosis, even in the presymptomatic stage, implying an etiologic link. However, the genetic mechanism underlying altered lipid levels in amyotrophic lateral sclerosis remains elusive. Therefore, in this study, we performed genetic correlation analysis, a cross-trait meta-analysis, tissue-specific enrichment analysis, and bidirectional two-sample Mendelian randomization analysis of European population to explore whether there is a genetic and causal relationship between lipids and amyotrophic lateral sclerosis. The effect of lipid-lowering drugs on amyotrophic lateral sclerosis was also evaluated using a drug target Mendelian randomization approach. The results showed a positive genetic correlation between amyotrophic lateral sclerosis and both high-density lipoprotein cholesterol and apolipoprotein A1 and identified 71 independent shared loci between amyotrophic lateral sclerosis and high-density lipoprotein cholesterol, as well as 55 independent shared loci between amyotrophic lateral sclerosis and apolipoprotein A1. These shared loci were enriched in the lipid metabolic pathway and the alcohol metabolic pathway. Further Mendelian randomization analysis targeting lipid-lowering drugs showed that single nucleotide polymorphisms within the ACLY and PCSK9 genes had a protective effect against amyotrophic lateral sclerosis risk by decreasing low-density lipoprotein cholesterol. The combination of ACLY and PCSK9 inhibitors has a greater protective effect on amyotrophic lateral sclerosis risk than that of PCSK9 inhibitors alone. In summary, there is a common genetic structure between lipids and amyotrophic lateral sclerosis. Mendelian randomization analysis supports an association between elevated blood lipids and the risk of developing amyotrophic lateral sclerosis, and the use of ACLY or PCSK9 inhibitors may improve disease prognosis.

To investigate the triglyceride-glucose index (TyG) as a predictor of glycaemic variability phenotypes in patients with type 2 diabetes mellitus (T2DM) using continuous glucose monitoring (CGM)-derived metrics, guiding personalized management. This cross-sectional study analysed 279 T2DM patients who underwent 14-day CGM monitering from community hospitals in Ningbo, China. Participants were stratified by TyG tertiles. Spectral clustering with complexity-invariant dynamic time warping identified distinct glycaemic variability glucotypes. Comprehensive CGM metrics spanning time-domain, frequency-domain, event-based, and circadian rhythm domains were evaluated. Multivariable logistic regression assessed associations between TyG and glycaemic phenotypes. Across TyG tertiles, we observed progressive increases in HbA1c (6.8 ± 1.5%, 7.2 ± 1.2%, 7.3 ± 1.3%, p = 0.011), fasting glucose (6.1 ± 1.1, 7.2 ± 1.6, 8.5 ± 2.8 mmol/L, p < 0.001), and triglycerides (0.9 ± 0.2, 1.4 ± 0.4, 2.7 ± 1.0 mmol/L, p < 0.001). Most notably, patients in the highest TyG tertile exhibited sustained hyperglycaemia with lower TIR (81.8% to 72.8 to 70.3%, p = 0.001), elevated TAR (15.9% vs. 29.1%, p < 0.001), and paradoxically reduced TBR (2.3% to 1.4% to 0.6%, p = 0.014) and CV (26.2% to 25.8% to 23.8%, p = 0.021). Frequency domain analysis demonstrated that elevated TyG was associated with high-level, low-frequency glucose oscillations. Higher TyG tertiles demonstrated increased prevalence of severe variability glucotype (32.3% to 53.8% to 63.4%), with TyG emerging as the strongest independent predictor (OR = 1.87, 95% CI: 1.42-2.50, p < 0.001). Elevated TyG identifies a distinct glycaemic phenotype characterized by high-level, low-frequency oscillations, sustained hyperglycaemia, and minimal hypoglycaemic risk. T2DM patients with high TyG levels may be appropriate candidates for more intensified glucose-lowering strategies.

To develop a tumor burden score (TBS)-based risk stratification system (RSS) for preoperative discrimination of lateral lymph node metastasis (LLNM) and compare it with conventional US and TBS-only models. This retrospective study included consecutive 404 adult patients diagnosed with PTC who underwent thyroid US and lateral lymph node dissection between January 2021 and October 2023. A prospective validation cohort was established with 174 patients enrolled from November 2023 to August 2024. Two novel tumor burden parameters, including D1/D2 and TBS score, were proposed. Logistic regression analyses were performed to assess associations between variables and LLNM in the training cohort. Features with p < 0.05 were incorporated into the LLNM RSS based on the Framingham risk score equation. Area under the receiver operating characteristic curve (AUC) was compared among the RSS, conventional US model, and TBS model in both cohorts. In both training and validation cohorts, the RSS showed improved performance for identifying LLNM compared with the TBS model (AUC, 0.809 vs. 0.703, p < 0.01 and AUC, 0.752 vs. 0.634, p = 0.03), and in the training cohort, the AUC of the RSS was superior compared with the conventional US model (AUC, 0.809 vs. 0.732, p < 0.01). Furthermore, we identified clustered punctate echogenic foci (PEF) distributed along the periphery of primary tumors as a robust independent predictor of LLNM (OR = 3.157, 95% CI: 1.520-6.557). The user-friendly RSS constructed based on the tumor burden concept outperformed conventional US and TBS models for distinguishing LLNM in PTC preoperatively. Question To develop a risk stratification system (RSS) for the prediction of lateral lymph node metastasis (LLNM) in papillary thyroid carcinoma (PTC), addressing the limitations of first-line ultrasound. Findings The tumor burden-based RSS achieved superior accuracy (AUC = 0.81) versus the conventional ultrasound-based model (AUC = 0.70), effectively evaluating the risk of LLNM preoperatively. Clinical relevance The RSS incorporating tumor burden parameters and peripheral punctate echogenic foci sign demonstrates strong predictive value for LLNM and enables risk-stratified surgical planning, avoiding unnecessary lymph node dissection in low-risk PTC while identifying occult metastases in high-risk cases.

CRISPR-based epigenome editing represents a programmable strategy to precisely modulate gene expression, holding immense promise for therapeutic applications. However, the large size of the dCas proteins substantially impedes the delivery via adeno-associated virus (AAV) vectors. Here, through iterative bioinformatics analysis, structure-guided predictions, and functional assays, we identified and characterized a novel miniature subtype V-M CRISPR-Cas12m from Pelomicrobium methylotrophicum. PmCas12m exhibited flexible 5'-YTN-3' PAM-dependent recognition and robust double-stranded DNA binding properties, while lacking DNA cleavage activity, thus positioning it as an ideal tool for epigenome editing. Cryogenic electron microscopy (cryo-EM) structures of PmCas12m unveiled its unique molecular mechanism of DNA binding facilitating interference. Guided by these structural insights, we employed deep mutational scanning (DMS) and protein engineering to develop xCas12m, a hypercompact variant with highly potent and specific epigenome editing capabilities in human cells. We further constructed the xCas12m-CRISPRoff platform in a single AAV vector, which achieved durable epigenetic silencing and effective inhibition of hepatitis B virus (HBV) infection in a mouse model. Collectively, these findings establish xCas12m as a versatile epigenome editing platform with transformative potential for treating diseases, paving the way for clinical translation of epigenetic therapies.

Osteosarcoma, the most common primary malignant bone tumour, presents significant treatment challenges due to its complex tumour microenvironment and the development of chemoresistance. This study employs single-cell transcriptomics to investigate chemotherapy-induced changes in osteosarcoma at both the cellular and molecular levels. Single-cell RNA sequencing data were analysed to identify cell subpopulations and their responses to chemotherapy. Differential gene expression and pathway enrichment analyses were performed to elucidate chemotherapy-induced changes. Additionally, we developed and validated a predictive model based on pyroptosis-related genes, named Pyroscore, using 101 different machine-learning algorithms. Chemotherapy led to an increased proportion of osteoclasts, endothelial cells, mesenchymal stem cells and pericytes, while decreasing T and NK cells, B cells, chondroblasts, monocytes and macrophages. Chemotherapy markedly upregulates the pyroptosis pathway in tumour cells, suggesting that chemotherapy induces programmed cell death in cancer cells through the activation of pyroptosis. Metabolic pathway analysis revealed significant inhibition of sulphur metabolism, starch and sucrose metabolism, pentose phosphate pathway, inositol phosphate metabolism, nitrogen metabolism and fatty acid metabolism. The Pyroscore model, which incorporates BAK1, CASP1, CASP5 and CASP6, demonstrated robust prognostic value across multiple data sets, with high scores correlating with improved survival outcomes. This study highlights the impact of chemotherapy on osteosarcoma cell subpopulations and the tumour microenvironment. The activation of the pyroptosis pathway and the development of the pyroscore prognostic model provide new insights into the mechanisms of chemotherapy response and potential therapeutic targets. These findings underscore the importance of personalized treatment strategies in improving outcomes for osteosarcoma patients.

Patients with peripheral T cell lymphoma (PTCL) who achieved tumor response with first-line standard therapy were at high risk of disease relapse. We explored golidocitinib (150 mg once daily) as maintenance therapy for this group of patients (JACKPOT26, NCT06511869). This study included two cohorts: patients achieving a complete response (Cohort 1 (CR), N = 30) and a partial response (Cohort 2 (PR), N = 18) during induction stage. All enrolled patients were transplant ineligible or did not have a transplant plan. All dosed patients were included in the efficacy and safety analysis. In Cohort 1, the 24-month disease free survival (DFS) rate was 74.2% with golidocitinib treatment. In nodal subtypes (AITL, NOS, ALK- ALCL), the 24-month DFS rate was 62.7%. In Cohort 2, median progression free survival (PFS) was 17.4 months, and 24-month PFS rate was 48.6%. Nine out of 18 patients with initial PR achieved complete response, leading to a complete response rate of 50.0%, and median duration of response of 23.9 months. The most common ≥grade 3 treatment-related treatment-emergent adverse events (TRAEs) were hematological adverse events in nature, including neutrophil count decreased (47.9%), white blood cell count decreased (31.3%), lymphocyte count decreased (14.6%) and leukopenia (12.5%). The majority of these TRAEs were reversible and clinically manageable. TRAEs leading to treatment interruption and discontinuation occurred in 60.4% and 10% of patients, respectively. No TRAEs leading to fatal outcomes were reported. This study suggests the potential of golidocitinib as maintenance therapy for patients with PTCL.

Lateral patellar compression syndrome (LPCS) is characterized by abnormal pressure elevation in the lateral patellofemoral joint. The histopathological basis of LPCS, particularly regarding microvascular and inflammatory changes in the lateral joint capsule and lateral retinaculum, remains unclear. This study aimed to systematically compare the histological characteristics of these tissues between patients with LPCS and controls to characterize tissue-level features associated with LPCS and to explore the potential implications of these findings for understanding pathophysiology and informing future research on surgical strategy selection. This retrospective case–control study included 26 patients with LPCS and 23 control patients (with meniscal injuries, patellar dislocation, patellofemoral osteoarthritis, or synovial cyst). Intraoperative tissue specimens of the lateral joint capsule and lateral retinaculum were obtained. Histological evaluation was performed using immunohistochemical staining for Factor Ⅷ (F8, microvessels), CD3 (T cells), CD20 (B cells), and CD68 (macrophages). Microvessel density and inflammatory cell density were quantified and compared between groups. Microvessel density was significantly higher in both the lateral joint capsule (48.84 ± 14.12/mm2 vs. 34.72 ± 15.40/mm2, P = 0.002) and the lateral retinaculum (27.50 ± 11.33/mm2 vs. 17.13 ± 9.26/mm2, P = 0.007) of patients with LPCS compared to controls, with large effect sizes (Cohen's d > 1.0). Within both groups, microvessel density was significantly higher in the joint capsule than in the retinaculum (both P < 0.001). In contrast, no statistically significant between-group differences were detected in the densities of CD3 + T cells, CD20 + B cells, or CD68 + macrophages in either tissue. However, in multivariate regression models adjusting for age and sex, the between-group differences in microvessel density were attenuated and no longer statistically significant; interaction terms were also not significant. Unadjusted analyses showed increased microvessel density in the lateral joint capsule and lateral retinaculum of patients with LPCS while no statistically significant between-group differences were detected in classical inflammatory cell densities (CD3 + , CD20 + , CD68 +) under the current immunohistochemical panel. Because the association with microvessel density was not statistically significant after adjustment for age and sex, these findings should be interpreted as hypothesis-generating and potentially influenced by demographic factors. Larger age- and sex-matched studies, ideally with clinical correlation, are needed to determine whether angiogenesis is an independent pathological feature of LPCS and whether it relates to outcomes.