Dysregulated RNA splicing is a well-recognized characteristic of colorectal cancer (CRC); however, its intricacies remain obscure, partly due to challenges in profiling full-length transcript variants at the single-cell level. Here, we employ high-depth long-read scRNA-seq to define the full-length transcriptome of colorectal epithelial cells in 12 CRC patients, revealing extensive isoform diversities and splicing alterations. Cancer cells exhibited increased transcript complexity, with widespread 3'-UTR shortening and reduced intron retention. Distinct splicing regulation patterns were observed between intrinsic-consensus molecular subtypes (iCMS), with iCMS3 displaying even higher splicing factor activities and more pronounced 3'-UTR shortening. Furthermore, we revealed substantial shifts in isoform usage that result in alterations of protein sequences from the same gene with distinct carcinogenic effects during tumorigenesis of CRC. Allele-specific expression analysis revealed dominant mutant allele expression in key oncogenes and tumor suppressors. Moreover, mutated PPIG was linked to widespread splicing dysregulation, and functional validation experiments confirmed its critical role in modulating RNA splicing and tumor-associated processes. Our findings highlight the transcriptomic plasticity in CRC and suggest novel candidate targets for splicing-based therapeutic strategies.
Humans ; Colorectal Neoplasms/metabolism* ; RNA Isoforms/metabolism* ; Single-Cell Analysis ; RNA Splicing ; Gene Expression Regulation, Neoplastic ; RNA, Neoplasm/metabolism* ; Transcriptome

Objective:To investigate the risk factors for new adjacent vertebral fracture after percutaneous kyphoplasty (PKP) in patients with osteoporotic vertebral compression fracture (OVCF) and their predictive efficacy.Methods:A retrospective cohort study was conducted to analyze the clinical data of 476 OVCF patients admitted to The Affiliated Wuxi People′s Hospital of Nanjing Medical University from January 2018 to December 2024, including 74 males and 402 females, aged 49-91 years [71(65, 79)years]. Among them, 397 patients underwent single-level PKP, while 79 received multi-level PKP. Surgical segments involved T 6 in 9 patients, T 7 in 9, T 8 in 14, T 9 in 12, T 10 in 9, T 11 in 50, T 12 in 110, L 1 in 173, L 2 in 77, L 3 in 46, L 4 in 31, and L 5 in 13. The patients were divided into adjacent vertebral fracture group ( n=55) and non-adjacent vertebral fracture group ( n=421) according to whether adjacent vertebral fracture was observed during the follow-up. The following data were collected in both groups: gender, age, body mass index (BMI), bone mineral density T-value, underlying diseases (hypertension, diabetes, coronary heart disease), prior cerebral infarction, history of OVCF, long-term glucocorticoid use, thoracolumbar fracture, number of operated vertebrae, cement injection approach (unilateral or bilateral), mean cement dose, postoperative vertebral height restoration rate, postoperative Cobb angle correction, postoperative thoracolumbar kyphosis angle correction, and cement distribution score. Univariate and multivariate Logistic stepwise regression analysis were performed to assess and identify independent risk factors for adjacent vertebral fracture in OVCF patients after PKP. Receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the risk factors′ predictive performance for adjacent vertebral fracture in OVCF patients after PKP. Results:Univariate analysis revealed significant differences in age, bone mineral density T-value, history of OVCF, long-term glucocorticoid use, number of operated vertebrae, and cement distribution score between the two groups ( P<0.05). The multivariate Logistic stepwise regression analysis showed that the bone mineral density T-value ( OR=0.68, 95% CI 0.48, 0.95, P<0.05) and cement distribution score ( OR=0.61, 95% CI 0.49, 0.76, P<0.01) were significantly correlated with new adjacent vertebral fractures after PKP. The ROC curve analysis showed that bone cement distribution score showed better predictive performance (AUC=0.72, 95% CI 0.64, 0.79), compared with bone mineral density T-value (AUC=0.62, 95% CI 0.54, 0.70), while the combined predictive performance of the two factors was the best (AUC=0.75, 95% CI 0.68, 0.81). Conclusions:Bone mineral density T-value and cement distribution score are independent risk factors for new adjacent vertebral fracture in OVCF patients after PKP. The predictive performance of cement distribution score is proved to be good and can be better in combination with bone mineral density T-value.

Objective:To investigate the clinical manifestations, treatment, and outcomes of Barth syndrome (BTHS).Methods:A retrospective analysis was conducted on 21 pediatric patients diagnosed with BTHS between January 2010 and December 2023 at Beijing Children′s Hospital, Beijing Anzhen Hospital, and Beijing JingDu Children′s Hospital. Clinical data including gender, age at onset, initial symptoms, clinical manifestations, personal history, family genetic history, and laboratory tests (neutrophil count, echocardiography, electrocardiogram and genetic testing) were reviewed.Results:All the 21 patients were male, with the age of onset at 4.1 (1.1, 9.3) months. Main clinical manifestations included heart failure (18 cases), neutropenia (16 cases), respiratory symptoms (15 cases), 3-methylpentenediuria (7 cases),develop retardation (8 cases), gastrointestinal symptoms (7 cases), fatigue and anorexia (6 cases), and recurrent infection (2 cases). Electrocardiogram abnormalities included ST changes (18 cases), flattened T wave and low voltage of limb leads (2 cases), and abnormal Q waves in lead Ⅰ and avL (1 case). Echocardiographic features showed increased trabeculation, interventricular septum and left ventricular wall thickening, and left ventricular enlargement with reduced ejection fraction. Genetic testing identified TAZ gene variations in all 21 patients: 11 missense mutations, 2 nonsense mutations, 2 frameshift mutations, 2 whole code mutations, 2 exon deletions, 1 splicing mutation, and 1 synonymous mutation. Fifteen mutations were maternally inherited, 2 were de novo, and 4 lacked verified variant origin.In terms of treatment, all 18 patients with heart failure received routine heart failure treatment, of whom 11 patients also received intravenous immunoglobulin and corticosteroids. After the follow-up of 91.0 (75.5, 109.5) months, 15 of the 18 patients showed restoration of cardiac function after 4.5 (3.0, 9.8) months of treatment, with one case of significant improvement, while 2 cases suddenly died.Conclusions:BTHS predominantly affects males with early onset, mainly characterized by abnormal cardiac structure and function, along with clinical features including fatigue, delayed growth and development, and neutropenia. Early diagnosis and intervention, including heart failure treatment, intravenous immunoglobulin, and corticosteroids, can lead to significant improvement in cardiac function, though sudden death remains a risk.

Objective To investigate the effect of tissue-resident memory T cells(TRM)on imiquimod-induced psoriatic-like skin lesions in mice,and to elucidate the underlying mechanisms of TRM involvement in this process.Methods Forty female BALB/c mice were procured and randomly allocated into four groups:ten in the blank control group,and thirty for the establishment of a psoriasis mouse model.Following successful modeling,the thirty mice were further randomized into three groups:the model control group,the methotrexate-treated group,and the imiquimod-treated group,with ten mice in each group.Mice in the blank control group and model control group were uniformly treated with Vaseline for intervention.The methotrexate group and the imiquimod group were treated with 62.5mg of 5％imiquimod cream.The methotrexate group was administered by gavage at a dose of 1 mg/kg,and the gavage volume of each group was 10 mL/kg.The model control group,blank group and imiquimod group were gavaged with the same volume of normal saline.Treatment was conducted over six consecutive days.Subsequently,comparisons were made across groups regarding the psoriasis area and severity index(PASI),histopathological findings,inflammatory cytokine levels,and TRM cell levels.Results(1)The imiquimod group exhibited signifi-cantly lower scores for erythema(2.54±0.32),skin thickening(2.59±0.25),and scaling(2.52±0.29)compared to the methotrexate group,model control group,and blank control group(P<0.05).Additionally,the methotrexate group demonstrated reduced scores for erythema,skin thickening,and scaling compared to the model control group(P<0.05).(2)Hematoxylin-eosin(HE)staining revealed that the epidermis in the methotrexate group became thin-ner,with fewer parakeratotic cells and increased hair follicles.Conversely,the imiquimod group displayed abnor-mal cell morphology and relatively thicker white skin after modeling.(3)The imiquimod group showed significantly lower levels of TNF-α(51.63±4.39 pg/mL),IL-1β(35.53±4.15 pg/mL),IFN-γ(23.43±3.41 pg/mL),and IL-23(15.24±2.95 pg/mL)compared to the methotrexate and model control groups(P<0.05).Similarly,the methotrexate group exhibited reduced levels of TNF-α,IL-1β,IFN-γ,and IL-23 compared to the model control group(P<0.05).(4)The imiquimod group had significantly lower levels of CD8+CD103+cells(15.39±2.31)than the methotrexate and model control groups(P<0.05).Furthermore,the methotrexate group demonstrated lower levels of CD8+CD103+cells compared to the model control group(P<0.05).Conclusion Miquimod induces heavier skin lesions,faster response,and more epidermal thickening in psoriasis like mice.CD8+CD103+TRM cells and inflammatory factors may be involved in the recurrence of psoriasis.

Objective To investigate the effect of tissue-resident memory T cells(TRM)on imiquimod-induced psoriatic-like skin lesions in mice,and to elucidate the underlying mechanisms of TRM involvement in this process.Methods Forty female BALB/c mice were procured and randomly allocated into four groups:ten in the blank control group,and thirty for the establishment of a psoriasis mouse model.Following successful modeling,the thirty mice were further randomized into three groups:the model control group,the methotrexate-treated group,and the imiquimod-treated group,with ten mice in each group.Mice in the blank control group and model control group were uniformly treated with Vaseline for intervention.The methotrexate group and the imiquimod group were treated with 62.5mg of 5％imiquimod cream.The methotrexate group was administered by gavage at a dose of 1 mg/kg,and the gavage volume of each group was 10 mL/kg.The model control group,blank group and imiquimod group were gavaged with the same volume of normal saline.Treatment was conducted over six consecutive days.Subsequently,comparisons were made across groups regarding the psoriasis area and severity index(PASI),histopathological findings,inflammatory cytokine levels,and TRM cell levels.Results(1)The imiquimod group exhibited signifi-cantly lower scores for erythema(2.54±0.32),skin thickening(2.59±0.25),and scaling(2.52±0.29)compared to the methotrexate group,model control group,and blank control group(P<0.05).Additionally,the methotrexate group demonstrated reduced scores for erythema,skin thickening,and scaling compared to the model control group(P<0.05).(2)Hematoxylin-eosin(HE)staining revealed that the epidermis in the methotrexate group became thin-ner,with fewer parakeratotic cells and increased hair follicles.Conversely,the imiquimod group displayed abnor-mal cell morphology and relatively thicker white skin after modeling.(3)The imiquimod group showed significantly lower levels of TNF-α(51.63±4.39 pg/mL),IL-1β(35.53±4.15 pg/mL),IFN-γ(23.43±3.41 pg/mL),and IL-23(15.24±2.95 pg/mL)compared to the methotrexate and model control groups(P<0.05).Similarly,the methotrexate group exhibited reduced levels of TNF-α,IL-1β,IFN-γ,and IL-23 compared to the model control group(P<0.05).(4)The imiquimod group had significantly lower levels of CD8+CD103+cells(15.39±2.31)than the methotrexate and model control groups(P<0.05).Furthermore,the methotrexate group demonstrated lower levels of CD8+CD103+cells compared to the model control group(P<0.05).Conclusion Miquimod induces heavier skin lesions,faster response,and more epidermal thickening in psoriasis like mice.CD8+CD103+TRM cells and inflammatory factors may be involved in the recurrence of psoriasis.

Objective:To investigate the risk factors for new adjacent vertebral fracture after percutaneous kyphoplasty (PKP) in patients with osteoporotic vertebral compression fracture (OVCF) and their predictive efficacy.Methods:A retrospective cohort study was conducted to analyze the clinical data of 476 OVCF patients admitted to The Affiliated Wuxi People′s Hospital of Nanjing Medical University from January 2018 to December 2024, including 74 males and 402 females, aged 49-91 years [71(65, 79)years]. Among them, 397 patients underwent single-level PKP, while 79 received multi-level PKP. Surgical segments involved T 6 in 9 patients, T 7 in 9, T 8 in 14, T 9 in 12, T 10 in 9, T 11 in 50, T 12 in 110, L 1 in 173, L 2 in 77, L 3 in 46, L 4 in 31, and L 5 in 13. The patients were divided into adjacent vertebral fracture group ( n=55) and non-adjacent vertebral fracture group ( n=421) according to whether adjacent vertebral fracture was observed during the follow-up. The following data were collected in both groups: gender, age, body mass index (BMI), bone mineral density T-value, underlying diseases (hypertension, diabetes, coronary heart disease), prior cerebral infarction, history of OVCF, long-term glucocorticoid use, thoracolumbar fracture, number of operated vertebrae, cement injection approach (unilateral or bilateral), mean cement dose, postoperative vertebral height restoration rate, postoperative Cobb angle correction, postoperative thoracolumbar kyphosis angle correction, and cement distribution score. Univariate and multivariate Logistic stepwise regression analysis were performed to assess and identify independent risk factors for adjacent vertebral fracture in OVCF patients after PKP. Receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the risk factors′ predictive performance for adjacent vertebral fracture in OVCF patients after PKP. Results:Univariate analysis revealed significant differences in age, bone mineral density T-value, history of OVCF, long-term glucocorticoid use, number of operated vertebrae, and cement distribution score between the two groups ( P<0.05). The multivariate Logistic stepwise regression analysis showed that the bone mineral density T-value ( OR=0.68, 95% CI 0.48, 0.95, P<0.05) and cement distribution score ( OR=0.61, 95% CI 0.49, 0.76, P<0.01) were significantly correlated with new adjacent vertebral fractures after PKP. The ROC curve analysis showed that bone cement distribution score showed better predictive performance (AUC=0.72, 95% CI 0.64, 0.79), compared with bone mineral density T-value (AUC=0.62, 95% CI 0.54, 0.70), while the combined predictive performance of the two factors was the best (AUC=0.75, 95% CI 0.68, 0.81). Conclusions:Bone mineral density T-value and cement distribution score are independent risk factors for new adjacent vertebral fracture in OVCF patients after PKP. The predictive performance of cement distribution score is proved to be good and can be better in combination with bone mineral density T-value.

Objective:To investigate the clinical manifestations, treatment, and outcomes of Barth syndrome (BTHS).Methods:A retrospective analysis was conducted on 21 pediatric patients diagnosed with BTHS between January 2010 and December 2023 at Beijing Children′s Hospital, Beijing Anzhen Hospital, and Beijing JingDu Children′s Hospital. Clinical data including gender, age at onset, initial symptoms, clinical manifestations, personal history, family genetic history, and laboratory tests (neutrophil count, echocardiography, electrocardiogram and genetic testing) were reviewed.Results:All the 21 patients were male, with the age of onset at 4.1 (1.1, 9.3) months. Main clinical manifestations included heart failure (18 cases), neutropenia (16 cases), respiratory symptoms (15 cases), 3-methylpentenediuria (7 cases),develop retardation (8 cases), gastrointestinal symptoms (7 cases), fatigue and anorexia (6 cases), and recurrent infection (2 cases). Electrocardiogram abnormalities included ST changes (18 cases), flattened T wave and low voltage of limb leads (2 cases), and abnormal Q waves in lead Ⅰ and avL (1 case). Echocardiographic features showed increased trabeculation, interventricular septum and left ventricular wall thickening, and left ventricular enlargement with reduced ejection fraction. Genetic testing identified TAZ gene variations in all 21 patients: 11 missense mutations, 2 nonsense mutations, 2 frameshift mutations, 2 whole code mutations, 2 exon deletions, 1 splicing mutation, and 1 synonymous mutation. Fifteen mutations were maternally inherited, 2 were de novo, and 4 lacked verified variant origin.In terms of treatment, all 18 patients with heart failure received routine heart failure treatment, of whom 11 patients also received intravenous immunoglobulin and corticosteroids. After the follow-up of 91.0 (75.5, 109.5) months, 15 of the 18 patients showed restoration of cardiac function after 4.5 (3.0, 9.8) months of treatment, with one case of significant improvement, while 2 cases suddenly died.Conclusions:BTHS predominantly affects males with early onset, mainly characterized by abnormal cardiac structure and function, along with clinical features including fatigue, delayed growth and development, and neutropenia. Early diagnosis and intervention, including heart failure treatment, intravenous immunoglobulin, and corticosteroids, can lead to significant improvement in cardiac function, though sudden death remains a risk.

Bone metastasis is a common complication of many advanced cancers,especially prostate cancer(85％),breast cancer(70％)and lung cancer(40％),which severely affects the quality of life of the pa-tients and increases its death risk.Macrophages are the essential immune cells in the human body that play a crucial role in the tumor microenvironment(TME),including phagocytosis or removal of cell debris,damaged or apoptotic cells.The phenotypic and functional transition of macrophages is a dynamic process.Under vari-ous stimulation conditions in TME,they can be transformed into two major subpopulations:M1 and M2.In TME,M1 macrophages exert anti-tumor effects and contribute to tumor immunity.M2 macrophages display a robust immunosuppressive phenotype,aiding in tumor cell proliferation and thereby promoting the occurrence of bone metastasis.This article reviews the interaction between macrophages and TME,and explores the role of macrophage polarization in bone metastasis of common cancers such as prostate cancer,breast cancer and lung cancer,and the influence in the occurrence and development of tumors.

Objective:To investigate the predictive factors for the outcome of patients with complete recanalization after endovascular treatment (EVT) for acute basilar artery occlusion (ABAO).Methods:Patients with ABAO underwent EVT at Jiangsu Provincial People's Hospital and Nanjing Pukou People's Hospital from January 2015 to December 2022 were included retrospectively. Complete recanalization was defined as a modified Thrombolysis in Cerebral Infarction (mTICI) grade 3 after EVT. The main outcome measure was the clinical outcome evaluated by the modified Rankin Scale at 90 days after onset. 0-2 points were defined as good outcome and >2 points were defined as poor outcome. The secondary outcome measure was death within 90 days after onset. Multivariate logistic regression analysis was used to determine the independent predictive factors for poor outcome and mortality. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of the independent predictive factors for poor outcome or mortality. Results:A total of 73 patients with completed recanalization after EVT for ABAO were enrolled, including 55 males (75.3%), aged 67.2±1.58 years, with the median baseline National Institutes of Health Stroke Scale (NIHSS) score 23, median baseline Basilar Artery on Computed Tomography Angiography (BATMAN) score 7, and median baseline posterior circulation Alberta Stroke Program Early CT Score (pc-ASPECTS) 8. After 90 days of onset, 34 patients (46.6%) had poor outcome and 16 (21.9%) died. Multivariate logistic regression analysis showed that higher baseline NIHSS score (odds ratio [ OR] 1.151, 95% confidence interval [ CI] 1.041-1.273; P=0.006), lower baseline pc-ASPECTS ( OR 0.096, 95% CI 0.024-0.386; P=0.001), lower baseline BATMAN score ( OR 0.394, 95% CI 0.162-0.961; P=0.041), and non-first-pass recanalization ( OR 5.011, 95% CI 1.675-23.343; P=0.016) were independently associated with the poor outcome. ROC curve analysis showed that the area under the curve for predicting poor outcome by combining these independent predictive factors was 0.966 (95% CI 0.930-0.964). Multivariate logistic regression analysis showed that older age ( OR 1.147, 95% CI 1.010-1.303; P=0.035), higher baseline NIHSS score ( OR 1.236, 95% CI 1.040-1.470; P=0.016), lower baseline pc-ASPECTS ( OR 0.011, 95% CI 0.002-0.249; P=0.015), and lower baseline BATMAN score ( OR 0.050, 95% CI 0.004-0.618; P=0.020) were independently associated with mortality within 90 days after onset. ROC curve analysis showed that the area under the curve for predicting mortality by combining these independent predictive factors was 0.948 (95% CI 0.899-0.997). Conclusion:For patients with ABAO who had complete recanalization after EVT, the baseline NIHSS score, baseline pc-ASPECTS, baseline BATMAN score, and non-first-pass recanalization are the independent predictive factors for poor outcome at 90 days after onset, while age, baseline NIHSS score, baseline pc-ASPECTS, and baseline BATMAN score are the independent predictive factors for mortality within 90 days after onset.

Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.