ObjectiveNeuroinflammation plays a crucial role in both the onset and progression of ischemic stroke, exerting a significant impact on the recovery of the central nervous system. Excessive neuroinflammation can lead to secondary neuronal damage, further exacerbating brain injury and impairing functional recovery. As a result, effectively modulating and reducing neuroinflammation in the brain has become a key therapeutic strategy for improving outcomes in ischemic stroke patients. Among various approaches, targeting immune regulation to control inflammation has gained increasing attention. This study aims to investigate the role of in vitro induced regulatory T cells (Treg cells) in suppressing neuroinflammation after ischemic stroke, as well as their potential therapeutic effects. By exploring the mechanisms through which Tregs exert their immunomodulatory functions, this research is expected to provide new insights into stroke treatment strategies. MethodsNaive CD4⁺ T cells were isolated from mouse spleens using a negative selection method to ensure high purity, and then they were induced in vitro to differentiate into Treg cells by adding specific cytokines. The anti-inflammatory effects and therapeutic potential of Treg cells transplantation in a mouse model of ischemic stroke was evaluated. In the middle cerebral artery occlusion (MCAO) model, after Treg cells transplantation, their ability to successfully migrate to the infarcted brain region and their impact on neuroinflammation levels were examined. To further investigate the role of Treg cells in stroke recovery, the changes in cytokine expression and their effects on immune cell interactions was analyzed. Additionally, infarct size and behavioral scores were measured to assess the neuroprotective effects of Treg cells. By integrating multiple indicators, the comprehensive evaluation of potential benefits of Treg cells in the treatment of ischemic stroke was performed. ResultsTreg cells significantly regulated the expression levels of both pro-inflammatory and anti-inflammatory cytokines in vitro and in vivo, effectively balancing the immune response and suppressing excessive inflammation. Additionally, Treg cells inhibited the activation and activity of inflammatory cells, thereby reducing neuroinflammation. In the MCAO mouse model, Treg cells were observed to accumulate in the infarcted brain region, where they significantly reduced the infarct size, demonstrating their neuroprotective effects. Furthermore, Treg cell therapy notably improved behavioral scores, suggesting its role in promoting functional recovery, and increased the survival rate of ischemic stroke mice, highlighting its potential as a promising therapeutic strategy for stroke treatment. ConclusionIn vitro induced Treg cells can effectively suppress neuroinflammation caused by ischemic stroke, demonstrating promising clinical application potential. By regulating the balance between pro-inflammatory and anti-inflammatory cytokines, Treg cells can inhibit immune responses in the nervous system, thereby reducing neuronal damage. Additionally, they can modulate the immune microenvironment, suppress the activation of inflammatory cells, and promote tissue repair. The therapeutic effects of Treg cells also include enhancing post-stroke recovery, improving behavioral outcomes, and increasing the survival rate of ischemic stroke mice. With their ability to suppress neuroinflammation, Treg cell therapy provides a novel and effective strategy for the treatment of ischemic stroke, offering broad application prospects in clinical immunotherapy and regenerative medicine.

Background:Ribosomal protein L22-like 1(RPL22L1)exerts regulatory effects on various malignant tumors such as lung cancer,prostate cancer,and cervical cancer.However,its role in colorectal cancer(CRC)remains unclear.Aims:To investigate the expression of RPL22L1 in CRC and its role in patients' prognosis and glucose metabolism of tumor cells.Methods:A total of 142 newly diagnosed CRC patients admitted to the Taizhou First People's Hospital from February 2022 to June 2024 were enrolled.The expression levels of RPL22L1 mRNA and protein were detected by quantitative real-time PCR and immunohistochemistry,respectively.The correlation between RPL22L1 expression and clinicopathological characteristics was analyzed.Kaplan-Meier survival analysis was used to evaluate the impact of RPL22L1 expression on the prognosis of CRC patients.RPL22L1 siRNA was transfected into SW480 cells to establish a low-expression cell model.Cell proliferation was assessed by CCK-8 assay,cell migration by Transwell chamber assay,and apoptosis by flow cytometry.Gene set enrichment analysis(GSEA)was performed to evaluate the effect of RPL22L1 on glucose metabolism of tumor cells.Results:The expression levels of RPL22L1 mRNA and protein were significantly higher in CRC tissues than in adjacent normal tissues(all P<0.05).The expression level of RPL22L1 mRNA was correlated with the TNM stage and carcinoembryonic antigen level of CRC(all P<0.05).Kaplan-Meier analysis showed that the cumulative survival rate of high RPL22L1 mRNA expression group was significantly lower than that of low-expression group(P=0.027).The expression level of RPL22L1 mRNA was significantly higher in SW480 cells than in normal intestinal epithelial cells(P<0.001).After inhibiting RPL22L1 expression,the proliferation and migration capacities of SW480 cells were significantly decreased(all P<0.05),the apoptosis rate was significantly increased(P=0.005),and the lactate level and relative glucose uptake level were significantly reduced(all P<0.05).GSEA indicated that RPL22L1 gene was associated with glycolysis/gluconeo-genesis(P=0.02).Conclusions:RPL22L1 is highly expressed in CRC and is associated with poor prognosis of patients,suggesting its potential as a molecular target for CRC therapy.Furthermore,RPL22L1 may promote the tumorigenesis and progression of CRC by modulating glucose metabolism.

Background and Aims:Long noncoding RNA(lncRNA)nuclear-enriched abundant transcript 1(NEAT1)is an oncogenic lncRNA that promotes the progression of various cancers through the competing endogenous RNA(ceRNA)mechanism.Using the TargetScan database,we previously identified binding sites between NEAT1 and microRNA-124-3p(miR-124-3p),as well as between miR-124-3p and catenin beta-1(CTNNB1).Therefore,this study was conducted to investigate the expression of NEAT1,miR-124-3p,and CTNNB1 in pancreatic cancer and their interactions affecting pancreatic cancer cell functions.Methods:A dual-luciferase reporter assay was used to validate the relationships among NEAT1,miR-124-3p,and CTNNB1.The expression levels of NEAT1 and miR-124-3p,as well as CTNNB1 protein expression,were detected in pancreatic cancer tissues and adjacent normal tissues,as well as in pancreatic cancer PANC-1 cells and normal pancreatic epithelial H6C7 cells.PANC-1 cells were transfected with NEAT1 siRNA alone or co-transfected with a miR-124-3p inhibitor.After transfection,changes in PANC-1 cell biological functions,epithelial-mesenchymal transition related protein expression,and tumor growth ability in mice were assessed.Results:The dual-luciferase reporter assay confirmed the targeting relationships between NEAT1 and miR-124-3p,as well as between miR-124-3p and CTNNB1.NEAT1 and CTNNB1 expression levels were significantly upregulated,while miR-124-3p expression was downregulated in pancreatic cancer tissues(vs.adjacent tissues)and in PANC-1 cells(vs.H6C7 cells)(all P<0.05).NEAT1 siRNA transfection led to decreased NEAT1 and CTNNB1 expression and increased miR-124-3p expression in PANC-1 cells.However,co-transfection with a miR-124-3p inhibitor suppressed the expression changes in miR-124-3p and CTNNB1(all P<0.05).NEAT1 siRNA transfection significantly reduced PANC-1 cell proliferation,migration,and invasion,while promoting apoptosis.Additionally,E-cadherin protein expression was upregulated,whereas N-cadherin and vimentin protein expression were downregulated.Tumor growth in mice was also significantly inhibited(all P<0.05).These changes were attenuated upon co-transfection with the miR-124-3p inhibitor(all P<0.05).Conclusion:NEAT1 may act as a ceRNA by competitively binding to miR-124-3p,thereby attenuating miR-124-3p-mediated inhibition of CTNNB1.This leads to CTNNB1 upregulation,ultimately promoting the malignant biological behavior of pancreatic cancer cells.

Background and Aims:Long noncoding RNA(lncRNA)nuclear-enriched abundant transcript 1(NEAT1)is an oncogenic lncRNA that promotes the progression of various cancers through the competing endogenous RNA(ceRNA)mechanism.Using the TargetScan database,we previously identified binding sites between NEAT1 and microRNA-124-3p(miR-124-3p),as well as between miR-124-3p and catenin beta-1(CTNNB1).Therefore,this study was conducted to investigate the expression of NEAT1,miR-124-3p,and CTNNB1 in pancreatic cancer and their interactions affecting pancreatic cancer cell functions.Methods:A dual-luciferase reporter assay was used to validate the relationships among NEAT1,miR-124-3p,and CTNNB1.The expression levels of NEAT1 and miR-124-3p,as well as CTNNB1 protein expression,were detected in pancreatic cancer tissues and adjacent normal tissues,as well as in pancreatic cancer PANC-1 cells and normal pancreatic epithelial H6C7 cells.PANC-1 cells were transfected with NEAT1 siRNA alone or co-transfected with a miR-124-3p inhibitor.After transfection,changes in PANC-1 cell biological functions,epithelial-mesenchymal transition related protein expression,and tumor growth ability in mice were assessed.Results:The dual-luciferase reporter assay confirmed the targeting relationships between NEAT1 and miR-124-3p,as well as between miR-124-3p and CTNNB1.NEAT1 and CTNNB1 expression levels were significantly upregulated,while miR-124-3p expression was downregulated in pancreatic cancer tissues(vs.adjacent tissues)and in PANC-1 cells(vs.H6C7 cells)(all P<0.05).NEAT1 siRNA transfection led to decreased NEAT1 and CTNNB1 expression and increased miR-124-3p expression in PANC-1 cells.However,co-transfection with a miR-124-3p inhibitor suppressed the expression changes in miR-124-3p and CTNNB1(all P<0.05).NEAT1 siRNA transfection significantly reduced PANC-1 cell proliferation,migration,and invasion,while promoting apoptosis.Additionally,E-cadherin protein expression was upregulated,whereas N-cadherin and vimentin protein expression were downregulated.Tumor growth in mice was also significantly inhibited(all P<0.05).These changes were attenuated upon co-transfection with the miR-124-3p inhibitor(all P<0.05).Conclusion:NEAT1 may act as a ceRNA by competitively binding to miR-124-3p,thereby attenuating miR-124-3p-mediated inhibition of CTNNB1.This leads to CTNNB1 upregulation,ultimately promoting the malignant biological behavior of pancreatic cancer cells.

Background:Ribosomal protein L22-like 1(RPL22L1)exerts regulatory effects on various malignant tumors such as lung cancer,prostate cancer,and cervical cancer.However,its role in colorectal cancer(CRC)remains unclear.Aims:To investigate the expression of RPL22L1 in CRC and its role in patients' prognosis and glucose metabolism of tumor cells.Methods:A total of 142 newly diagnosed CRC patients admitted to the Taizhou First People's Hospital from February 2022 to June 2024 were enrolled.The expression levels of RPL22L1 mRNA and protein were detected by quantitative real-time PCR and immunohistochemistry,respectively.The correlation between RPL22L1 expression and clinicopathological characteristics was analyzed.Kaplan-Meier survival analysis was used to evaluate the impact of RPL22L1 expression on the prognosis of CRC patients.RPL22L1 siRNA was transfected into SW480 cells to establish a low-expression cell model.Cell proliferation was assessed by CCK-8 assay,cell migration by Transwell chamber assay,and apoptosis by flow cytometry.Gene set enrichment analysis(GSEA)was performed to evaluate the effect of RPL22L1 on glucose metabolism of tumor cells.Results:The expression levels of RPL22L1 mRNA and protein were significantly higher in CRC tissues than in adjacent normal tissues(all P<0.05).The expression level of RPL22L1 mRNA was correlated with the TNM stage and carcinoembryonic antigen level of CRC(all P<0.05).Kaplan-Meier analysis showed that the cumulative survival rate of high RPL22L1 mRNA expression group was significantly lower than that of low-expression group(P=0.027).The expression level of RPL22L1 mRNA was significantly higher in SW480 cells than in normal intestinal epithelial cells(P<0.001).After inhibiting RPL22L1 expression,the proliferation and migration capacities of SW480 cells were significantly decreased(all P<0.05),the apoptosis rate was significantly increased(P=0.005),and the lactate level and relative glucose uptake level were significantly reduced(all P<0.05).GSEA indicated that RPL22L1 gene was associated with glycolysis/gluconeo-genesis(P=0.02).Conclusions:RPL22L1 is highly expressed in CRC and is associated with poor prognosis of patients,suggesting its potential as a molecular target for CRC therapy.Furthermore,RPL22L1 may promote the tumorigenesis and progression of CRC by modulating glucose metabolism.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Background::Breast cancer (BC) risk-stratification tools for Asian women that are highly accurate and can provide improved interpretation ability are lacking. We aimed to develop risk-stratification models to predict long- and short-term BC risk among Chinese women and to simultaneously rank potential non-experimental risk factors.Methods::The Breast Cancer Cohort Study in Chinese Women, a large ongoing prospective dynamic cohort study, includes 122,058 women aged 25-70 years old from the eastern part of China. We developed multiple machine-learning risk prediction models using parametric models (penalized logistic regression, bootstrap, and ensemble learning), which were the short-term ensemble penalized logistic regression (EPLR) risk prediction model and the ensemble penalized long-term (EPLT) risk prediction model to estimate BC risk. The models were assessed based on calibration and discrimination, and following this assessment, they were externally validated in new study participants from 2017 to 2020.Results::The AUC values of the short-term EPLR risk prediction model were 0.800 for the internal validation and 0.751 for the external validation set. For the long-term EPLT risk prediction model, the area under the receiver operating characteristic curve was 0.692 and 0.760 in internal and external validations, respectively. The net reclassification improvement index of the EPLT relative to the Gail and the Han Chinese Breast Cancer Prediction Model (HCBCP) models for external validation was 0.193 and 0.233, respectively, indicating that the EPLT model has higher classification accuracy.Conclusions::We developed the EPLR and EPLT models to screen populations with a high risk of developing BC. These can serve as useful tools to aid in risk-stratified screening and BC prevention.

The present analysis was based on the iteration of the 2023 WHO Essential Medicines Model List(23rd list)and Essential Medicines Model List for Children(9th list).In a methodological fashion,the adjustment procedure of the basic drug list and the materials required to submit the application were sorted out.Subsequently,a comprehensive synthesis of predominant features characterizing the 2023 edition of the EML and EMLc are summarized(offered).Notably,this edition boasts the highest count of medicines ever included featuring extensive inclusion of innovative drugs to fill the gap,select medicines for children carefully,and the emphasizing on clinical benefit evidence and public health affordability.It is suggested that China should update the NEML according to the national conditions and the experience of WHO as soon as possible,pay attention to the difference and connection between the list of essential medicines and the list of medical insurance,and play a meticulous role for establishing effective linkages between two lists.Such measures are envisaged to meet the basic drug needs,and ensure the accessibility and affordability of drugs,thereby optimizing the allocation of health resources.

Background:Colorectal cancer is a common malignant tumor in the digestive tract.The existing treatment methods and drugs have disadvantages such as significant toxicity and side effects,so there is an urgent need to develop new drugs with high efficiency and low toxicity.Aims:To investigate the potential mechanism of Nodosin in inducing apoptosis of colorectal cancer cells.Methods:Colorectal cancer cell lines SW480 and HCT116 were cultured with 0,5,10 and 20 μmol/L Nodosin for 24 hours,or cultured with 10 μmol/L Nodosin for 0,12,24 and 48 hours.The cell viability was measured by CCK-8 assay,and the protein expressions of PARP1,mTOR,p-mTOR and LC3BⅡ were detected by Western blotting.After pretreated with the autophagy inhibitor 3-methyladenine(3-MA)5 mmol/L for 4 hours,SW480 and HCT116 cells were cultured with 10 μmol/L Nodosin for 24 hours.The protein expressions of PARP1 and LC3BⅡ were detected by Western blotting.Results:Nodosin could decrease the cell viability of SW480 and HCT116 cells in a dose-and time-dependent manner,increase the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ,and inhibit the protein expression of p-mTOR.Pretreatment of the autophagy inhibitor 3-MA could inhibit the expressions of apoptosis-related protein cleaved PARP1 and autophagy-related protein LC3BⅡ induced by Nodosin.Conclusions:Nodosin can induce apoptosis of colorectal cancer cells by inhibiting mTOR pathway and activating autophagy.