This study aims to investigate the protective effects and mechanisms of polysaccharide extract PCP1 from Polygonatum cyrtonema in ameliorating cerebral ischemia-reperfusion(I/R) injury in rats through modulation of the Toll-like receptor 4(TLR4)/NOD-like receptor protein 3(NLRP3) signaling pathway. In vivo, SD rats were randomly divided into the sham group, model group, PCP1 group, nimodipine(NMDP) group, and TLR4 signaling inhibitor(TAK-242) group. A middle cerebral artery occlusion/reperfusion(MCAO/R) model was established, and neurological deficit scores and infarct size were evaluated 24 hours after reperfusion. Hematoxylin-eosin(HE) and Nissl staining were used to observe pathological changes in ischemic brain tissue. Transmission electron microscopy(TEM) assessed ultrastructural damage in cortical neurons. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-18(IL-18), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), and nitric oxide(NO) in serum. Immunofluorescence was used to analyze the expression of TLR4 and NLRP3 proteins. In vitro, a BV2 microglial cell oxygen-glucose deprivation/reperfusion(OGD/R) model was established, and cells were divided into the control, OGD/R, PCP1, TAK-242, and PCP1 + TLR4 activator lipopolysaccharide(LPS) groups. The CCK-8 assay evaluated BV2 cell viability, and ELISA determined NO release. Western blot was used to analyze the expression of TLR4, NLRP3, and downstream pathway-related proteins. The results indicated that, compared with the model group, PCP1 significantly reduced neurological deficit scores, infarct size, ischemic tissue pathology, cortical cell damage, and the levels of inflammatory factors IL-1β, IL-6, IL-18, TNF-α, and NO(P<0.01). It also elevated IL-10 levels(P<0.01) and decreased the expression of TLR4 and NLRP3 proteins(P<0.05, P<0.01). Moreover, in vitro results showed that, compared with the OGD/R group, PCP1 significantly improved BV2 cell viability(P<0.05, P<0.01), reduced cell NO levels induced by OGD/R(P<0.01), and inhibited the expression of TLR4-related inflammatory pathway proteins, including TLR4, myeloid differentiation factor 88(MyD88), tumor necrosis factor receptor-associated factor 6(TRAF6), phosphorylated nuclear factor-kappaB dimer RelA(p-p65)/nuclear factor-kappaB dimer RelA(p65), NLRP3, cleaved-caspase-1, apoptosis-associated speck-like protein(ASC), GSDMD-N, IL-1β, and IL-18(P<0.05, P<0.01). The protective effects of PCP1 were reversed by LPS stimulation. In conclusion, PCP1 ameliorates cerebral I/R injury by modulating the TLR4/NLRP3 signaling pathway, exerting anti-inflammatory and anti-pyroptotic effects.
Animals ; Toll-Like Receptor 4/genetics* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Rats, Sprague-Dawley ; Rats ; Reperfusion Injury/genetics* ; Male ; Signal Transduction/drug effects* ; Polysaccharides/isolation & purification* ; Polygonatum/chemistry* ; Brain Ischemia/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Humans

Ovarian cancer is the most lethal malignancy affecting the female reproductive system. Pharmacological inhibitors targeting CDK4/6 have demonstrated promising efficacy across various cancer types. However, their clinical benefits in ovarian cancer patients fall short of expectations, with only a subset of patients experiencing these advantageous effects. This study aims to provide further clinical and biological evidence for antineoplastic effects of a CDK4/6 inhibitor (TQB4616) in ovarian cancer and explore underlying mechanisms involved. Patient-derived ovarian cancer organoid models were established to evaluate the effectiveness of TQB3616. Potential key genes related to TQB3616 sensitivity were identified through RNA-seq analysis, and TRIM4 was selected as a candidate gene for further investigation. Subsequently, co-immunoprecipitation and GST pull-down assays confirmed that TRIM4 binds to hnRNPDL and promotes its ubiquitination through RING and B-box domains. RIP assay demonstrated that hnRNPDL binded to CDKN2C isoform 2 and suppressed its expression by alternative splicing. Finally, in vivo studies confirmed that the addition of siTRIM4 significantly improved the effectiveness of TQB3616. Overall, our findings suggest that TRIM4 modulates ubiquitin-mediated degradation of hnRNPDL and weakens sensitivity to CDK4/6 inhibitors in ovarian cancer treatment. TRIM4 may serve as a valuable biomarker for predicting sensitivity to CDK4/6 inhibitors in ovarian cancer.
Humans ; Female ; Ovarian Neoplasms/pathology* ; Animals ; Tripartite Motif Proteins/genetics* ; Mice ; Cyclin-Dependent Kinase 4/antagonists & inhibitors* ; Cell Line, Tumor ; Cyclin-Dependent Kinase 6/antagonists & inhibitors* ; Protein Kinase Inhibitors/pharmacology* ; Ubiquitin/metabolism* ; Xenograft Model Antitumor Assays ; Ubiquitination ; Antineoplastic Agents/pharmacology*

Crigler-Najjar syndrome(CNS)is an autosomal recessive genetic disorder caused by mutations in the UGT1A1 gene.This author retrospectively analyzes the clinical data of one patient presenting with postnatal hyperbilirubinemia,who was genetically diagnosed with CNS.Her clinical course,genetic characteristics,diagnostic and therapeutic approaches are discussed to enhance clinicians'understanding of this condition.The patient exhibited recurrent jaundice due to elevated unconjugated bilirubin(UCB)levels since birth.During this period,the gallstones were identified and surgically removed,yet her bilirubin levels did not improve.In March 2024,the patient presented to our hospital and underwent genetic testing via a third-party facility.The results revealed two risk variants in the UGT1A1 gene.After one week of oral phenobarbital administration,her UCB levels decreased by approximately 47%,confirming a diagnosis of CNS type Ⅱ.The patient is currently under active follow-up with regular monitoring of bilirubin levels.For the infants and children presenting with persistent,fluctuating,non-hemolytic,and non-hepatitic hyperbilirubinemia since birth,early genetic testing should be prioritized to establish the definitive diagnosis.

The ubiquitin-proteasome system (UPS) is responsible for protein degradation in both normal and pathological states. E3 ligases selectively attach ubiquitin to specific substrates, which is essential for regulating cellular homeostasis. The function of E3 ligases has been associated with a variety of diseases, such as cancer and cardiovascular disease. The discovery of E3 ligands can help regulate E3 ligases, thus expanding new ideas for disease treatment. Targeted protein degradation (TPD) drugs, including proteolysis targeting chimera (PROTAC), have become increasingly popular in recent years due to their dependence on E3 ligands. In this paper, we review the discovery techniques of E3 ligands, including activity-based protein mapping, fragment-based drug discovery, and library-based methods, and briefly introduce the protein interaction detection techniques involved in the ligand discovery techniques, in the hope of providing certain ideas for the future discovery of E3 ligands as well as the treatment of diseases.

Objective To investigate the differentially expressed genes(DEGs)and their molecular interactions in unstable carotid atherosclerotic plaques.Methods Gene expression datasets related to carotid atherosclerotic plaques(GSE41571,GSE118481,and E-MTAB-2055)were downloaded from Gene Expression Omnibus(GEO)and European Bioinformatics Institute(EBI)ArrayExpress databases.The co-regulated DEGs in at least two datasets of unstable carotid plaques were merged and analyzed using Gene Ontology Biological Process(GO-BP),Kyoto Encyclopedia of Genes and Genomes(KEGG),Protein-Protein Interaction(PPI)Networks and subnetwork analysis,relationships between miRNAs/transcription factors and target genes,and drug-gene interaction database.Quantitative real-time PCR(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to detect the expression levels of some DEGs in carotid plaques and plasma from 58 patients with carotid atherosclerosis.Results GO enrichment analysis showed that DEGs in unstable carotid atherosclerotic plaques were mainly enriched in genes related to inflammatory response and extracellular matrix structure genes.KEGG enrichment analysis indicated that upregulated DEGs in unstable carotid plaques were enriched in extracellular matrix receptor(ECM-receptor)interaction,PI3K-Akt,Hippo and transforming growth factor-β(TGF-β)signaling pathways,while downregulated DEGs were primarily enriched in lysosomes,phagosomes,and chemokines processes.PPI network analysis suggested that COL1A2,COL4A2,insulin-like growth factor binding protein 6(IGFBP6),COL4A5,C1QA,CXCL10,CXCL2,CXCR4,and CSF1R may play important roles in PPI networks.Prediction of drug-gene interactions revealed that CSF1R had the most drug interaction,CXCL2 was most antagonized by drugs,and IGFBP6 was most activated by drugs.qRT-PCR showed that the expression level of IGFBP6 in unstable carotid plaques group was significantly lower than that in stable carotid plaques group(P<0.001).ELISA results showed that plasma concentration of IGFBP6 in unstable carotid plaques group was significantly lower than that in stable carotid plaques group(P<0.0001).Receiver operating characteristic(ROC)suggested that the area under the curve(AUC)for plasma IGFBP6 levels to identify unstable plaques was 0.894(95%CI 0.810-0.977),with a cutoff value of 142.08 ng/ml.Conclusion IGFBP6 may become an important biomarker for predicting unstable carotid atherosclerotic plaques.

Objective:To investigate the predictive value of Glasgow coma scale-pupil (GCS-P) score combined with arterial lactate level on 30-day mortality in patients with spontaneous intracerebral hemorrhage (sICH).Methods:Patients with sICH attended to the emergency department of the Affiliated Hospital of North China University of Science and Technology from January 2021 to September 2023 were respectively collected. The patients were divided into survival group and death group according to survival outcome at 30 d after the onset of the disease. The data of the patients between the two groups were compared, including basic data, vital signs on admission to the hospital, pupils status, laboratory indexes, emergency Glasgow coma scale (GCS) scores, and other clinical data. The binary logistic regression analysis was used to analyze the risk factors affecting the poor prognosis of sICH patients, and LASSO regression analysis was used for further validation. The receiver operating characteristic (ROC) curves were plotted to analyze the predictive value of the emergency GCS-P score combined with the lactate level score on the mortality rates of sICH patients.Results:A total of 512 patients were included, 235 in the survival group and 277 in the death group. The admission heart rate, blood glucose, arterial lactate level, and the proportion of patients with hypertension and stroke were significantly higher in the death group than in the survival group, and the GCS score and emergency GCS-P score were significantly lower than those in the survival group (all P<0.05). For the mortality analysis of sICH patients at different levels, the mortality rate of patients with very severe GCS-P was significantly higher than that of patients with severe disease ( P<0.05), However, there was no significant difference in GCS between the two groups ( P>0.05). Binary logistic regression analysis showed that emergency GCS-P score ( OR=1.400, 95% CI: 1.297-1.512, P<0.001) and arterial lactate level ( OR=0.674, 95% CI: 0.567-0.800, P<0.001) were predicted factors for the prognosis of patients with sICH. ROC curve analysis showed that the area under the curve of GCS, GCS-P, arterial lactate, and GCS-P combined with arterial lactate to predict the prognosis of patients was 0.748, 0.783, 0.718, and 0.819, respectively. Among them, GCS-P score combined with arterial lactate had the best prediction effect. Conclusion:GCS-P combined with arterial lactate levels could be used to predict the prognosis of sICH patients

Objective:To study the diagnostic value of artificial intelligence B-ultrasound image computer-aided diagnosis system (hereinafter referred to as intelligent ultrasound system) in adult goiter.Methods:In June 2022 and March 2023, two phases of thyroid disease survey were carried out in 4 cities in Anhui Province. One village was selected in each city, and 250 adults were selected as survey subjects in each village. Adult bilateral thyroid area was scanned by both intelligent ultrasound system and conventional ultrasound scanning equipment, and the effectiveness of intelligent ultrasound system in the diagnosis of goiter was analyzed based on the results of conventional ultrasound examination. Receiver operating characteristic (ROC) curve was drawn, and Kappa test was used to analyze the consistency between intelligent ultrasound system and conventional ultrasound examination in the diagnosis of goiter. At the same time, Spearman correlation analysis and Bland-Altman method were used to evaluate the consistency of the two methods in measuring thyroid volume.Results:After screening and removing outliers and missing values, a total of 910 adults were included, including 253 males (27.80%) and 657 females (72.20%). The age was (45.92 ± 10.20) years old, ranging from 18 to 60 years old. The sensitivity, specificity, and accuracy of the intelligent ultrasound system for diagnosing adult goiter were 80.00%, 99.67%, and 99.56%, respectively. The area under the ROC curve (AUC) was 0.996, which was consistent with the results of conventional ultrasound examination for diagnosing goiter ( κ = 0.67, P < 0.001). After controlling for variables such as gender, thyroid function, and thyroid nodules, the intelligent ultrasound system showed good consistency with conventional ultrasound examination in the diagnosis of goiter in females, adults with thyroid dysfunction, and adults without thyroid nodules ( κ = 0.66, 0.80, 0.80, P < 0.001). The consistency in the diagnosis of goiter in adults with thyroid nodules was moderate ( κ = 0.56, P < 0.001). Spearman correlation analysis showed a highly positive correlation between the measurement results of adult thyroid volume by intelligent ultrasound system and conventional ultrasound examination ( r = 0.88, P < 0.001). The Bland-Altman method results showed that only 4.62% (42/910) of points in adults were outside the 95% consistency limit, indicating good consistency between intelligent ultrasound system and conventional ultrasound examination in measuring thyroid volume (< 5%). The proportion of points outside the 95% consistency limit in males, adults with thyroid dysfunction, and adults with thyroid nodules was 6.72% (17/253), 5.83% (12/206), and 6.45% (12/186), respectively. Conclusions:The intelligent ultrasound system has certain diagnostic value for adult goiter and has good consistency with conventional ultrasound examination for thyroid volume measurement. However, the accuracy of diagnosis for males and adults with thyroid nodules still needs to be improved.

Objective To compare the safety and efficacy of endovascular treatment(EVT)transradial approach and transfemoral approach in patients with acute ischemic stroke with large vessel occlusion in the posterior circulation.Methods Patients with acute ischemic stroke with large vessel occlusion in the posterior circulation admitted to the Stroke Center of Linyi People's Hospital,Shandong Second Medical University from February 2022 to April 2024 were retrospectively recruited.The baseline and clinical data were collected,including age,sex,past medical history(hypertension,diabetes mellitus,hyperlipidemia,atrial fibrillation,coronary artery disease,myocardial infarction and stroke),smoking,blood pressure at admission,National Institutes of Health stroke scale(NIHSS)score at admission,modified Rankin scale(mRS)score before the onset of stroke,Alberta stroke program early CT score(ASPECTS),whether intravenous thrombolysis was performed,volume of core infarct zone,volume of hypoperfused zone,occlusion location,etiology of stroke,time indexes(including time from onset to door,time from door to arterial puncture,arterial puncture to recanalization,and time from onset to revascularization),anesthesia modality,EVT first-line strategy,number of passes and whether with vascular access conversion.Clinical data included 90-day postoperative mRS scores,postoperative puncture site complications in 24 hours,symptomatic intracranial hemorrhage at 72 hours postoperatively,modified thrombolysis in cerebral infarction(mTICI)grade at the last angiography introperative and length of stay.The groups were categorized into transradial(TRA)and transfemoral(TFA)groups according to the final access route for EVT.The baseline and clinical data of the two groups were compared.Results A total of 129 patients underwent EVT with acute ischemic stroke with large vessel occlusion in the posterior circulation were finally included,including 47 patients in TRA group and 82 patients in TFA group.The differences about sex,age,past medical history,smoking,blood pressure at admission,NIHSS score at admission,mRS score before the onset of the stroke,ASPECTS,whether intravenous thrombolysis was performed,volume of core infarct zone,volume of hypoperfused zone,occlusion location,etiology of stroke,anesthesia modality,EVT first-line strategy,number of passes,mTICI grade at the last angiography introperative and other aspects were not statistically significant(all P＞0.05).The good prognosis rate at 90 d after surgery(53.2％[25/47]vs.48.8％[40/82],P=0.630),distribution of mRS scores at 90d postoperatively(P=0.991),all-cause mortality at 90 days after surgery(27.7％[13/47]vs.28.0％[23/82],P=1.000),rate of good recanalization(mTICI grade≥2b)on intraoperative last angiogram(97.9％[46/47]vs.95.1％[78/82],P=0.436),rate of complication of puncture site(4.3％[2/47]vs.12.2％[10/82],P=0.209)between the two groups were not statistically significant.The time from door to arterial puncture was significantly longer in the TRA group than in the TFA group,and the difference between the two groups was statistically significant(122.00[95.00,153.00]min vs.105.00[80.25,118.00]min,Z=-2.937,P=0.03);average length of stay in the TFA group was significantly longer than that of the TRA group,and the difference between the two groups was statistically significant(6.00[4.95,7.05]d vs.7.00[6.95,8.88]d,Z=-2.573,P=0.010).Conclusions Patients who underwent EVT via radial or femoral artery approach with acute ischemic stroke with large vessel occlusion in the posterior circulation had similar safety and efficacy,and the number of days of hospitalization was shorter via the TRA.However,more large prospective randomized controlled clinical trials are needed to validate the results of this study.

Objective To investigate the predictive value of Glasgow coma scale(GCS)-Full Out-line of UnResponsiveness(FOUR)scale(GCS-F)score and arterial lactate level for prognosis of intracerebral hemorrhage(ICH)in elderly patients.Methods A total of 278 elderly ICH patients admitted to Neurosurgical Intensive Care Unit of the Affiliated Hospital of North China Universi-ty of Science and Technology from January 2022 to January 2024 were retrospectively recruited.According score at 90 d after onset,95 of them were assigned into good prognosis group and 183 into poor prognosis group.Basic information,vital signs at admission,pupils,laboratory indica-tors,GCS score and other clinical data were compared between the two groups.Binary logistic re-gression analysis was used to analyze the risk factors of poor prognosis in elderly ICH patients.ROC curve was drawn to analyze the prognostic value of GCS-F score(including eye opening re-sponse and limb movement in GCS,and brainstem response in FOUR scale)combined with lac-tate level in elderly ICH patients.Results Significantly higher blood pressure,elevated levels of blood glucose,creatinine and arterial lactic acid,increased percentages of supratentorial and sub-tentorial bleeding,and larger blood loss,while obviously lower GCS score and GCS-F score were observed in the poor prognosis group than the good prognosis group(P＜0.05,P＜0.01).In the analyses for poor prognosis rate of ICH patients at different levels,the poor prognosis rate of se-vere GCS-F patients was significantly higher than that of patients with same level of GCS score(P＜0.05).Binary logistic regression analysis showed that GPS-F score(OR=1.762,95％CI:1.507-2.061,P=0.000)and arterial lactate level(OR=0.536,95％CI:0.385-0.746,P=0.000)were risk factors for 90-day poor prognosis in elderly ICH patients.ROC curve analysis indicated that the AUC value of arterial lactate,GCS score and GCS-F score in predicting 90-day poor prog-nosis in elderly ICH patients was 0.713(95％CI:0.650-0.775),0.827(95％CI:0.774-0.880)and 0.876(95％CI:0.828-0.925),respectively,and the AUC value of GCS-F score combined with arterial lactic acid was 0.919(95％CI:0.886-0.953),which was significantly higher than that of GCS score,GCS-F score and arterial lactic acid alone(P＜0.01).Conclusion GCS-F score has better performance than GCS score in predicting 90-day poor prognosis in elderly ICH pa-tients,especially for severe patients.The GPS-F score combined with arterial lactate level shows the best predictive effectiveness for elderly ICH patients.

OBJECTIVE To establish a determination method for the related substances of LPM3480392, a novel Gi protein-biased opioid receptor(MOR) agonist. METHODS The separation was carried out with Waters Symmetry Shield RP18 (150 mm×4.6 mm, 3.5 μm) by gradient elution method, using a mixture of 0.002 5 mol·L–1 sodium 1-octanesulfonate monohydrate in 0.01 mol·L–1 potassium dihydrogen phosphate-water solution(containing 0.1% triethylamine, adjusted pH to 2.50 with phosphate acid) and acetonitrile as the mobile phase at a flow rate of 1.0 mL·min–1 and the UV detection wavelength was set at 210 nm. RESULTS The chromatographic peaks of LPM3480392 and impurity A, impurity B, impurity C, impurity E and impurity F could be completely separated, the linear relationship of LPM3480392 was good in 0.064 9−5.191 2 μg·mL–1, while impurity A, impurity B, impurity C, impurity E and impurity F showed good linear relationship within 0.066 6−7.610 4 μg·mL–1, 0.166 0−3.794 0 μg·mL–1, 0.209 2−4.463 2 μg·mL–1, 0.167 9−7.672 6 μg·mL–1 and 0.016 4−7.505 7 μg·mL–1, respectively. The recovery rate was within 93.0%−103.2%. CONCLUSION The method is suitable for the determination of related substances in LPM3480392, and can provide valuable reference for the follow-up research and development of LPM3480392.