Hypoxic-ischemic (HI) brain damage poses a high risk of death or lifelong disability, yet effective treatments remain elusive. Here, we demonstrated that miR-100-5p levels in the lesioned cortex increased after HI insult in neonatal mice. Knockdown of miR-100-5p expression in the brain attenuated brain injury and promoted functional recovery, through inhibiting the cleaved-caspase-3 level, microglia activation, and the release of proinflammation cytokines following HI injury. Engineered extracellular vesicles (EVs) containing neuron-targeting rabies virus glycoprotein (RVG) and miR-100-5p antagonists (RVG-EVs-Antagomir) selectively targeted brain lesions and reduced miR-100-5p levels after intranasal delivery. Both pre- and post-HI administration showed therapeutic benefits. Mechanistically, we identified protein phosphatase 3 catalytic subunit alpha (Ppp3ca) as a novel candidate target gene of miR-100-5p, inhibiting c-Fos expression and neuronal apoptosis following HI insult. In conclusion, our non-invasive method using engineered EVs to deliver miR-100-5p antagomirs to the brain significantly improves functional recovery after HI injury by targeting Ppp3ca to suppress neuronal apoptosis.
Animals ; MicroRNAs/metabolism* ; Extracellular Vesicles/metabolism* ; Mice ; Recovery of Function/physiology* ; Hypoxia-Ischemia, Brain/therapy* ; Mice, Inbred C57BL ; Antagomirs/administration & dosage* ; Male ; Animals, Newborn ; Apoptosis/drug effects* ; Brain Injuries/metabolism* ; Glycoproteins ; Peptide Fragments ; Viral Proteins

Objective To analyze the effects of fluid overload(FO)on clinical outcomes of extremely preterm infants with hemody-namically significant patent ductus arteriosus(hsPDA).Methods This retrospective study analyzed data from neonates ≤ 32 weeks ges-tational age(GA)admitted to the Neonatal Intensive Care Unit(NICU)of the Affiliated Hospital of Xuzhou Medical University between November 2021 and July 2024.Infants were categorized into hsPDA and non-hsPDA groups based on the presence or absence of hsPDA.General clinical data and daily FO situation in the first week after birth of the two groups were compared,and the risk factors affecting hsPDA were obtained by Logistic regression analysis.Using the first day of postnatal FO as the test variable and hsPDA as the state varia-ble,receiver operator characteristic(ROC)curve was drawn to calculate the cut-off value of FO,and the cut-off value was divided into groups to analyze the relationship between the first day FO and clinical outcome of very preterm infants.Results A total of 153 extremely preterm infants(GA≤32 weeks)were enrolled,including 110 in the hsPDA group and 43 in the non-HSPDA group.Univariate analysis showed that there were statistically significant differences in birth weight,1-minute Apgar score,postnatal antibiotic use time,parenteral nutrition time,invasive mechanical ventilation time,non-invasive mechanical ventilation time,hospital days,day 1,day 2 and day 4 FO between the two groups(P＜0.05).Multivariate Logistic regression analysis showed that invasive mechanical ventilation time and day 1 FO were risk factors of hsPDA.The ROC curve was drawn with the first day FO as the test variable and hsPDA as the state variable.The area under the curve was 0.903,the sensitivity and specificity were 79.1％and 89.1％,respectively,and the cut-off value of FO on the first day after birth was 8.78％.According to this cut-off value,107 cases were divided into FO≤8.78％group and 46 cases were FO＞8.78％group.Univariate analysis showed that FO＞8.78％group had lower Apgar score at 1 minute,longer invasive mechani-cal ventilation time,and higher incidence of moderate to severe bronchopulmonary dysplasia(BPD)and grade 3 to 4 intraventricular hem-orrhage(IVH).Logistic regression analysis showed that FO＞8.78％on the first day after birth was a risk factor for moderate to severe BPD and grade 3-4 IVH.Conclusion FO＞8.78％on the first day after birth of very preterm infants increased the incidence of moder-ate to severe BPD and grade 3-4 IVH.

BACKGROUND:The self-renewal and multi-directional differentiation of pluripotent stem cells possess the potential to revolutionize people's understanding of biology,medicine,development,and disease.Stem cells play an important role in the early stage of embryonic development,and the study of them could be beneficial to understanding of the basic principles of biological development and tissue or organ formation,exploring the potential mechanisms of various diseases,studying the repair and regeneration of damaged tissues or organs,and promoting drug discovery and personalized treatment. OBJECTIVE:To review the research progress of pluripotent stem cells,summarize and categorize the fundamental types of pluripotent stem cells,and elucidate the lineage situations of various types of pluripotent stem cells in common mammals. METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched systematically,with the keywords"pluripotent stem cells;embryonic stem cells;induced pluripotent stem cells;expanded potential stem cells;livestock pluripotent stem cells"in English and Chinese.The 99 articles related to mammalian pluripotent stem cells were systematically screened according to inclusion and exclusion criteria,and then reviewed. RESULTS AND CONCLUSION:(1)According to classical theory in mouse embryonic stem cell research,the pluripotent state of stem cells is divided into two forms:na?ve and primed.Na?ve state corresponds to the inner cell mass of pre-implantation embryos before attachment to the uterine wall,while primed state corresponds to the epiblast after implantation.These two states exhibit significant differences in epigenetic features,transcriptional activity,external signal dependency,and metabolic phenotype.It is later discovered that there is an intermediate state between na?ve and primed called formative pluripotency.Therefore,the pluripotency of pluripotent stem cells is a continuous developmental process rather than a unique cell state.(2)In addition to obtaining pluripotent stem cells from the inner cell mass,there are various methods and lineages for acquiring pluripotent stem cells,including embryonic germ cells established using primitive germ cells from mouse embryos,induced pluripotent stem cells created by the dedifferentiation of adult mouse and human fibroblasts with four factors—Oct3/4,Sox2,c-Myc,and Klf4;embryonic stem cell-like cell lines cultured from somatic cell nuclear transfer,parthenogenesis,neonatal or adult testicular or ovarian tissue,very small embryonic-like stem cells derived from various adult tissues and expanded pluripotent stem cells derived from pre-implantation stages.These pluripotent stem cells all share the common characteristics of continuous self-renewal,expressing core pluripotency factors and possessing the ability to differentiate into the three primary germ layers.(3)Currently,pluripotent stem cells are being used for disease modeling to study the mechanisms of various diseases and develop new drugs.Simultaneously,scientists are attempting to use pluripotent stem cells to cultivate various tissues and organs,offering new possibilities for regenerative medicine and transplantation.However,the clinical application of pluripotent stem cells faces safety challenges,including issues of cell mutations and immune rejection.Continual improvement in the methods of generating pluripotent stem cells will make them safer and more efficient for clinical applications.(4)Based on the methods of obtaining and lineage establishment of pluripotent stem cells in mice and humans,various types of pluripotent stem cells have been established in livestock,including embryonic stem cells,induced pluripotent stem cells,germ lineages of pluripotent stem cells,and expanded potential stem cells.Research on livestock pluripotent stem cells opens up new avenues for animal reproduction,breeding,genetic engineering,disease modeling,drug screening,and the conservation of endangered wildlife.

Objective To analyze the effects of fluid overload(FO)on clinical outcomes of extremely preterm infants with hemody-namically significant patent ductus arteriosus(hsPDA).Methods This retrospective study analyzed data from neonates ≤ 32 weeks ges-tational age(GA)admitted to the Neonatal Intensive Care Unit(NICU)of the Affiliated Hospital of Xuzhou Medical University between November 2021 and July 2024.Infants were categorized into hsPDA and non-hsPDA groups based on the presence or absence of hsPDA.General clinical data and daily FO situation in the first week after birth of the two groups were compared,and the risk factors affecting hsPDA were obtained by Logistic regression analysis.Using the first day of postnatal FO as the test variable and hsPDA as the state varia-ble,receiver operator characteristic(ROC)curve was drawn to calculate the cut-off value of FO,and the cut-off value was divided into groups to analyze the relationship between the first day FO and clinical outcome of very preterm infants.Results A total of 153 extremely preterm infants(GA≤32 weeks)were enrolled,including 110 in the hsPDA group and 43 in the non-HSPDA group.Univariate analysis showed that there were statistically significant differences in birth weight,1-minute Apgar score,postnatal antibiotic use time,parenteral nutrition time,invasive mechanical ventilation time,non-invasive mechanical ventilation time,hospital days,day 1,day 2 and day 4 FO between the two groups(P＜0.05).Multivariate Logistic regression analysis showed that invasive mechanical ventilation time and day 1 FO were risk factors of hsPDA.The ROC curve was drawn with the first day FO as the test variable and hsPDA as the state variable.The area under the curve was 0.903,the sensitivity and specificity were 79.1％and 89.1％,respectively,and the cut-off value of FO on the first day after birth was 8.78％.According to this cut-off value,107 cases were divided into FO≤8.78％group and 46 cases were FO＞8.78％group.Univariate analysis showed that FO＞8.78％group had lower Apgar score at 1 minute,longer invasive mechani-cal ventilation time,and higher incidence of moderate to severe bronchopulmonary dysplasia(BPD)and grade 3 to 4 intraventricular hem-orrhage(IVH).Logistic regression analysis showed that FO＞8.78％on the first day after birth was a risk factor for moderate to severe BPD and grade 3-4 IVH.Conclusion FO＞8.78％on the first day after birth of very preterm infants increased the incidence of moder-ate to severe BPD and grade 3-4 IVH.

Objective:To observe the changes of peripheral blood T cell subsets and serum tumor necrosis factor (TNF)-α and interleukin (IL)-6 contents in narcolepsy type 1 (NT1) patients and their correlations with narcolepsy, and provide basis for finding the biological markers of narcolepsy.Methods:A retrospective analysis was performed. From March 2022 to December 2023, 23 patients with NT1 admitted to Epilepsy and Sleep Disorder Center, Second Affiliated Hospital of Harbin Medical University and 23 healthy controls underwent physical examination of nervous system in our center were enrolled. T lymphocyte subsets CD4 + and CD8 + in peripheral blood were calculated by flow cytometry. Serum TNF-α and IL-6 contents were detected by enzyme-linked immunosorbent assay. Multivariate Logistic regression was used to determine the correlations of NT1 with CD4 + T lymphocyte count and IL-6 and TNF-α contents, and diagnostic values of CD4 + T lymphocyte and TNF-α in NT1 were evaluated via area under receiver operating characteristics (ROC) curve. Results:Compared with the healthy controls, the NT1 patients had significantly increased peripheral blood CD4 + T lymphocyte count ([820.61±316.87] /μL vs. [1121.04±387.47] /μL), and significantly higher serum TNF-α and IL-6 contents ([39.97±10.64] pg/mL vs. [57.01±19.92] pg/mL; [22.50±6.09] pg/mL vs. [33.66±17.28] pg/mL, P<0.05). No significant difference in peripheral blood CD8 + T lymphocyte count was noted between the 2 groups ([668.65±276.45] pg/mL vs. [592.52±217.78] pg/mL, P>0.05). Multivariate Logistic regression showed that CD4 + T lymphocyte count and serum TNF-α content were independent risk factors for NT1 ( OR=1.004, 95% CI: 1.001-1.006, P=0.007; OR=1.133, 95% CI: 1.032-1.243, P=0.009). Area under ROC curve of the two combined indexes was 0.881(95% CI: 0.784-0.977, P=0.001), enjoying sensitivity of 0.783 and specificity of 0.870. Conclusion:Combination of peripheral blood CD4 + T lymphocyte count and serum TNF-α content has high diagnostic performance in predicting NT1.

Objective:To investigate the efficacy, safety and possible mechanisms of celecoxib as an adjunctive treatment for schizophrenia.Methods:90 schizophrenic inpatients at the second affiliated hospital of Xinxiang Medical College from April 2019 to October 2020 were recruited and randomly assigned to a placebo group or the celecoxib adjunctive treatment group using a random number table. In the placebo group, 46 patients (29 males, 17 females; aged 21-34, mean age 27.46±6.50 years) completed a 6-week follow-up. In the celecoxib group, 44 patients (32 males, 12 females; aged 21-39, mean age 30.52±8.69 years) completed a 6-week follow-up. The Positive and Negative Syndrome Scale (PANSS) was used to assess psychiatric symptoms in both groups. Changes in PANSS score at the end of the treatment were compared to evaluate the efficacy of celecoxib. Metabolic indicators such as weight, body mass index, waist circumference and plasm glucolipid, as well as cardiovascular indicators like blood pressure, electrocardiogram and routine blood tests, and adverse events were collected for the safety evaluation. Serum tumor necrosis factor-α (TNF-α), Interleukin-4 (IL-4) and interferon-γ (IFN-γ) were also tested. Pearson correlation analysis was used to explore the relationship between cytokine levels, PANSS score, PANSS reduction rate [(pre-treatment score-post-treatment score)/pre-treatment score×100%], and the safety measurements in the two groups, analyzing the role of inflammation in celecoxib adjunctive therapy.Results:The change of PANSS positive score at the end of the 6th week was significantly higher in the celecoxib adjuvant treatment group than in the placebo group (-8.00±6.12 vs -4.78±5.19, H=-0.55, P=0.009). The weight changes, body mass index, total cholesterol, and triglycerides over 6 weeks were significantly lower in the celecoxib group compared to the placebo group ( F=-7.37, -7.30, 2.56, -2.54; all P<0.05). No serious adverse events were found in celecoxib adjuvant therapy. In the placebo group, baseline TNF-α levels were positively correlated with baseline negative symptoms and PANSS reduction rate ( r=0.260 and 0.330, both P<0.05), and negatively correlated with the 6-week weight ( r=-0.311, P<0.05); baseline IL-4 levels were positively correlated with the 6-week PANSS total score and the 6-week PANSS negative score ( r=0.320 and 0.397, both P<0.05), and negatively correlated with PANSS reduction rate and 6-week blood glucose ( r=-0.316 and -0.331, both P<0.05); Six-week IFN-γ levels were negatively correlated with low-density lipoprotein levels ( r=-0.306, P<0.05). And no such correlation was found in celecoxib adjuvant group. Conclusion:Celecoxib adjunctive therapy can improve positive symptoms of schizophrenia without causing adverse reactions. Inflammatory state is related to schizophrenia symptoms, treatment efficacy and metabolic abnormalities.

Objective:To explore the predictive ability of systemic immune-inflammation index（SII）and prognostic nutrition index（PNI）for early-onset sepsis in preterm infants.Methods:Seventy preterm infants of 28 to 32 weeks，who were born in the Affiliated Hospital of Xuzhou Medical University from January 2019 to December 2022, and transferred to neonatal intensive care unit within 1 h conforming to EOS diagnostic criteria were selected as the EOS group，and 1∶1 matched non-infected preterm infants hospitalized during the same period were selected as control group.Relevant data were collected to compare the differences regarding clinical data，blood routine indicators，C-reactive protein（CRP），serum albumin levels（ALB），SII and PNI between two groups.The ability of SII and PNI to predict EOS was evaluated by Logistic regression analysis and receiver operating characteristic（ROC）curve.Results:Compared with control group,the EOS group had lower 1-minute and 5-minute Apgar scores,higher rates of cesarean section delivery and tracheal intubation,as well as higher rates of suppurative meningitis,bronchopulmonary dysplasia,retinopathy of prematurity and intracranial hemorrhage.The levels of blood routine parameters,ALB,SII and PNI in the EOS group were lower than those in control group,while CRP was increased.The differences were all statistically significant（ P<0.05）.Multivariate Logistic regression analysis showed that tracheal intubation，CRP，SII and PNI were independently influential factors of EOS（ P<0.05）.ROC curve analysis showed that the areas under curve of SII，PNI，CRP and SII combined with PNI were 0.808（95% CI 0.730-0.886），0.792（95% CI 0.718-0.865），0.633（95% CI 0.541-0.725）and 0.866（95% CI 0.803-0.929），the sensitivity were 74.3%，64.3%，42.9%，78.6%，and the specificity were 88.6%，82.9%，81.4%，90.0%,respectively.The cut-off values of SII，PNI and CRP were 221.36，38.65 and 0.80 mg/L,respectively. Conclusion:SII and PNI have a certain predictive value for EOS in preterm infants，and their combined diagnosis efficiency is more stronger.

Objective To develop and validate a risk prediction model based on early platelet-related parameters for bronchopulmonary dysplasia(BPD)in neonates admitted to the neonatal intensive care unit(NICU),and to facilitate early identification and intervention in high-risk populations.Methods Clinical data of 291 preterm infants with a gestational age(GA)≤32 weeks or a birth weight(BW)＜1 500 g,admitted to the NICU,were retrospectively analyzed.Out of these,214 cases were selected as the modeling group.This group was further categorized into the BPD group(n=76)and the non-BPD group(n=138),based on whether they required oxygen therapy at 28 days post-birth.Perinatal data,platelet-related parameters and other indicators between the two groups.Univariate and multivariate Logistic regression analyses were conducted to identify BPD risk factors,followed by the construction of a nomogram.An additional cohort of 105 preterm infants with GA≤32 weeks or BW＜1 500 g,were used to validate the model.This cohort was divided into the BPD group(n=43)and the non-BPD(n=62)group.Receiver operating characteristic(ROC)curve and calibration curve were used to internally verify the efficiency of the prediction model.Results The Logistic regression analysis identified GA,BW,Apgar score at 5 minutes≤7,invasive ventilation,platelet count(PLT)and mean platelet volume(MPV)as significant factors in the model(P＜0.05).The constructed nomogram was formulated using R language,and the areas under the ROC curve(AUC)for the three models were 0.908,0.931 and 0.918,respectively(P＜0.05).The verification group was verified by Bootstrap.The calibration curve showed a good fit.The internal validation AUC values of the three models were 0.877,0.890 and 0.886,respectively.Conclusion GA,BW,invasive ventilation,Apgar score at 5 minutes≤7,MPV and PLT are key risk factors for BPD onset.The risk prediction model based on these indicators can effectively predict BPD,providing clinicians with a valuable tool for early detection and intervention in the development of BPD.

Background::While type 2 diabetes mellitus (T2DM) is considered a putative causal risk factor for coronary artery disease (CAD), the intrinsic link underlying T2DM and CAD is not fully understood. We aimed to highlight the importance of integrated care targeting both diseases by investigating the phenotypic and genetic relationships between T2DM and CAD.Methods::We evaluated phenotypic associations using data from the United Kingdom Biobank ( N = 472,050). We investigated genetic relationships by leveraging genomic data conducted in European ancestry for T2DM, with and without adjustment for body mass index (BMI) (T2DM: Ncase/ Ncontrol = 74,124/824,006; T2DM adjusted for BMI [T2DM adjBMI]: Ncase/ Ncontrol = 50,409/523,897) and for CAD ( Ncase/ Ncontrol = 181,522/984,168). We performed additional analyses using genomic data conducted in multiancestry individuals for T2DM ( Ncase/ Ncontrol = 180,834/1,159,055). Results::Observational analysis suggested a bidirectional relationship between T2DM and CAD (T2DM→CAD: hazard ratio [HR] = 2.12, 95% confidence interval [CI]: 2.01–2.24; CAD→T2DM: HR = 1.72, 95% CI: 1.63–1.81). A positive overall genetic correlation between T2DM and CAD was observed ( rg = 0.39, P = 1.43 × 10 -75), which was largely independent of BMI (T2DM adjBMI–CAD: rg = 0.31, P = 1.20 × 10 –36). This was corroborated by six local signals, among which 9p21.3 showed the strongest genetic correlation. Cross-trait meta-analysis replicated 101 previously reported loci and discovered six novel pleiotropic loci. Mendelian randomization analysis supported a bidirectional causal relationship (T2DM→CAD: odds ratio [OR] = 1.13, 95% CI: 1.11-1.16; CAD→T2DM: OR = 1.12, 95% CI: 1.07-1.18), which was confirmed in multiancestry individuals (T2DM→CAD: OR = 1.13, 95% CI: 1.10-1.16; CAD→T2DM: OR = 1.08, 95% CI: 1.04-1.13). This bidirectional relationship was significantly mediated by systolic blood pressure and intake of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, with mediation proportions of 54.1% (95% CI: 24.9-83.4%) and 90.4% (95% CI: 29.3-151.5%), respectively. Conclusion::Our observational and genetic analyses demonstrated an intrinsic bidirectional relationship between T2DM and CAD and clarified the biological mechanisms underlying this relationship.

Objective:To investigate the efficacy, safety and possible mechanisms of celecoxib as an adjunctive treatment for schizophrenia.Methods:90 schizophrenic inpatients at the second affiliated hospital of Xinxiang Medical College from April 2019 to October 2020 were recruited and randomly assigned to a placebo group or the celecoxib adjunctive treatment group using a random number table. In the placebo group, 46 patients (29 males, 17 females; aged 21-34, mean age 27.46±6.50 years) completed a 6-week follow-up. In the celecoxib group, 44 patients (32 males, 12 females; aged 21-39, mean age 30.52±8.69 years) completed a 6-week follow-up. The Positive and Negative Syndrome Scale (PANSS) was used to assess psychiatric symptoms in both groups. Changes in PANSS score at the end of the treatment were compared to evaluate the efficacy of celecoxib. Metabolic indicators such as weight, body mass index, waist circumference and plasm glucolipid, as well as cardiovascular indicators like blood pressure, electrocardiogram and routine blood tests, and adverse events were collected for the safety evaluation. Serum tumor necrosis factor-α (TNF-α), Interleukin-4 (IL-4) and interferon-γ (IFN-γ) were also tested. Pearson correlation analysis was used to explore the relationship between cytokine levels, PANSS score, PANSS reduction rate [(pre-treatment score-post-treatment score)/pre-treatment score×100%], and the safety measurements in the two groups, analyzing the role of inflammation in celecoxib adjunctive therapy.Results:The change of PANSS positive score at the end of the 6th week was significantly higher in the celecoxib adjuvant treatment group than in the placebo group (-8.00±6.12 vs -4.78±5.19, H=-0.55, P=0.009). The weight changes, body mass index, total cholesterol, and triglycerides over 6 weeks were significantly lower in the celecoxib group compared to the placebo group ( F=-7.37, -7.30, 2.56, -2.54; all P<0.05). No serious adverse events were found in celecoxib adjuvant therapy. In the placebo group, baseline TNF-α levels were positively correlated with baseline negative symptoms and PANSS reduction rate ( r=0.260 and 0.330, both P<0.05), and negatively correlated with the 6-week weight ( r=-0.311, P<0.05); baseline IL-4 levels were positively correlated with the 6-week PANSS total score and the 6-week PANSS negative score ( r=0.320 and 0.397, both P<0.05), and negatively correlated with PANSS reduction rate and 6-week blood glucose ( r=-0.316 and -0.331, both P<0.05); Six-week IFN-γ levels were negatively correlated with low-density lipoprotein levels ( r=-0.306, P<0.05). And no such correlation was found in celecoxib adjuvant group. Conclusion:Celecoxib adjunctive therapy can improve positive symptoms of schizophrenia without causing adverse reactions. Inflammatory state is related to schizophrenia symptoms, treatment efficacy and metabolic abnormalities.