Objective To investigate the role of NF-κB in apoptosis of immortalized neural progenitor cells (INPCs) . Methods INPCs were cultured in 6-well plates and were randomly divided into 5 groups ( n = 6 each) : group I was not transfected with any plasmid (group INPC); group Ⅱ was transfected with control plasmid (group INPC/CMV); group Ⅲ was transfected with plasmid RcCMV-p50 (group INPC/p50); group Ⅳ was transfected with plasmid RcCMV-p65 (group INPC/p65) and group V was transfected with plasmid RcCMV-p50 and RcCMV-p65 (group INPC/p50p65). Group INPC/CMV ( H ), INPC/p50 (Ⅲ) and INPC/p65 (Ⅳ) were screened by G418, and the positive clones were then cultured for 3-4 weeks. The transcription of p50 mRNA or p6S mRNA was detected by RT-PCR. The NF-κB activity was measured by luciferase reporter gene assay. The cell apoptosis was measured by annexin V/PI staining. In group INPC/p50p65 and group INPC/p65, the cultured positive clone was transiently transfected with plasmid RcCMV-p50. Two days after transfection, the same measurement was performed in group INPC/pS0p65 and the other groups. Results The expression of p50 mRNA was significantly increased in group INPC/p50 and INPC/p50p65 as compared with the other groups ( P < 0.05) . The expression of p65 mRNA, the NF-κB activity and the apoptotic rate were significantly increased in group INPC/p65 and INPC/p50p65 as compared with the other groups ( P < 0.05). Conclusion Enhanced NF-κB activity can increase immortalized neural progenitor cell apoptosis.

ObjectiveTo investigate the expression of APC-Cdh1 protein after cerebral ischemia-reperfusion injury.Methods60 male Sprague-Dawley rats were randomly divided into Sham-operated group(SH) and ischemia-reperfusion group(IR). The rats of ischemia-reperfusion groups were induced by four-vessel occlusion (4-VO). At different times after injury, the expression of APC-Cdh1 of rat hippocampus was observed by Western blotting and immunohistochemistry.ResultsCompared with sham-operated group, the expression of Cdh1 protein significantly decreased 1 day and increased obviously 3 days, but decreased again 7 days after injury in ischemia-reperfusion group. The immuno-staining showed that APC-Cdh1 was highly cerebral cortex and hippocampus in ischemia-reperfusion group. ConclusionAPC-Cdh1 may be involved in the central nervous system injury.

BACKGROUND:Cdh1 has been shown to express in rat hippocampus and cortex in a large number. Moreover, in vitro test demonstrated that Cdh1 expression was higher in neurons than in neural stem cel s, which possibly associated with the differentiation of neural stem cel s into neurons. However, the effects of anaphase promoting complex Cdh1 on ischemic neuronal damage remain unclear.
OBJECTIVE:To investigate the expression of Cdh1 and its downstream substrate in primary cultured neurons with oxygen-glucose deprivation. METHODS:Primary neurons from cortex of postnatal 24-hour rat pups were cultured in vitro, and identified by immunofluorescence staining. The oxygen-and glucose-deprived models were established by three gas incubator fil ed with nitrogen in sugar-free Earle’s solution. After 1 hour of hypoxia, reoxygenation was conducted. Real-time fluorescent quantitative PCR was used to detect the mRNA expression of Cdh1 and its downstream substrates Skp2, Cyclin B1 before hypoxia, 6 hours, 1, 3, 7 days after oxygen glucose deprivation. RESULTS AND CONCLUSION:After oxygen glucose deprivation, the expression of Cdh1 and Cyclin B1 in primary neurons was increased (P<0.05), while Skp2 expression was decreased (P<0.05). Above data indicated that Cdh1 expression in neurons increased after oxygen-glucose deprivation. It may degrade Skp2 and participate in hypoxic neuronal apoptosis by ubiquitination.

Objective: To determine whether noncontrast computed tomography (NCCT) models based on multivariable, radiomics features, and machine learning (ML) algorithms could further improve the discrimination of early hematoma expansion (HE) in patients with spontaneous intracerebral hemorrhage (sICH). Materials and Methods: We retrospectively reviewed 261 patients with sICH who underwent initial NCCT within 6 hours of ictus and follow-up CT within 24 hours after initial NCCT, between April 2011 and March 2019. The clinical characteristics, imaging signs and radiomics features extracted from the initial NCCT images were used to construct models to discriminate early HE. A clinical-radiologic model was constructed using a multivariate logistic regression (LR) analysis. Radiomics models, a radiomics-radiologic model, and a combined model were constructed in the training cohort (n = 182) and independently verified in the validation cohort (n = 79). Receiver operating characteristic analysis and the area under the curve (AUC) were used to evaluate the discriminative power. Results: The AUC of the clinical-radiologic model for discriminating early HE was 0.766. The AUCs of the radiomics model for discriminating early HE built using the LR algorithm in the training and validation cohorts were 0.926 and 0.850, respectively.The AUCs of the radiomics-radiologic model in the training and validation cohorts were 0.946 and 0.867, respectively. The AUCs of the combined model in the training and validation cohorts were 0.960 and 0.867, respectively. Conclusion NCCT models based on multivariable, radiomics features and ML algorithm could improve the discrimination of early HE. The combined model was the best recommended model to identify sICH patients at risk of early HE.

Objective:To investigate the predictive value of the model based on soluble T cell immunogloblulin and mucin domain-containing protein 3 (sTIM3) for the progression of severe acute pancreatitis (SAP) in patients with acute pancreatitis (AP).Methods:A retrospective cohort study was conducted. The AP patients admitted to Changzhou First People's Hospital and Changzhou Second People's Hospital from June 1, 2020 to June 30, 2022 were enrolled. Mild AP (MAP) and moderately severe AP (MSAP) patients were classified as non-SAP group, and SAP patients were classified as SAP group according to the progression of AP patients during hospitalization. The basic data, blood biological indicators, serum sTIM3 level, bedside index for severity in acute pancreatitis (BISAP), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, modified computed tomography severity index (MCTSI) score within 48 hours of admission, and prognosis indicators were collected. Multivariate Logistic regression analysis was conducted to analyze the risk factors of the progression of SAP in patients with AP during hospitalization. Based on the results of multivariate analysis and the best parameters selected based on the minimal Akaike information criterion (AIC), the SAP prediction model based on sTIM3 was constructed. The receive operator characteristic curve (ROC curve) was plotted to analyze the predictive efficacy of the model.Results:A total of 99 AP patients were enrolled, 80 patients in the non-SAP group and 19 patients in the SAP group. Compared with the non-SAP group, body mass index (BMI), drinking history ratio, heart rate (HR), respiration rate (RR), white blood cell count (WBC), red blood cell count (RBC), C-reactive protein (CRP), alanine aminotransferase (ALT), serum creatinine (SCr), procalcitonin (PCT), interleukin-6 (IL-6), sTIM3, BISAP score, APACHEⅡ score and MCTSI score were significantly increased, and pulse oxygen saturation (SpO 2), direct bilirubin (DBil) and IL-10 were significantly decreased. The length of intensive care unit (ICU) stay and total length of hospital stay of patients in the SAP group were significantly longer than those in the non-SAP group [length of ICU stay (days): 1.0 (0, 1.5) vs. 0 (0, 0), total length of hospital stay (days): 17.11±9.39 vs. 8.40±3.08, both P < 0.01]. Multivariate Logistic regression analysis showed that HR [odds ratio ( OR) = 1.059, 95% confidence interval (95% CI) was 1.010-1.110, P = 0.017], DBil ( OR = 0.981, 95% CI was 0.950-0.997, P = 0.043), and sTIM3 ( OR = 1.002, 95% CI was 1.001-1.003, P = 0.027) were independent risk factors for predicting the progression of SAP in patients with AP, and the SAP prediction model based on sTIM3 was constructed: Logit( P) = -14.602+0.187×BMI+0.057×HR+0.006×CRP-0.020×DBil+0.002×sTIM3. ROC curve analysis showed that among the aforementioned single factor quantitative indicators, IL-6 was the most effective in predicting the progression of AP patients to SAP during hospitalization, but the predictive performance of prediction model based on the sTIM3 was significantly better than IL-6 [area under the ROC curve (AUC) and 95% CI: 0.957 (0.913-1.000) vs. 0.902 (0.845-0.958), P < 0.05]. Conclusion:The model based on serum sTIM3 demonstrated good predictive value for the progression of SAP in patients with AP.

Cordyceps sinensis(C.sinensis)is a widely used and highly valuable traditional Chinese medicine.Several dipeptides have been detected in C.sinensis,but current scientific knowledge of its chemical makeup remains limited.In this study,an improved approach that integrates offline two-dimensional liquid chromatography(2D LC)separation,precursor ion list,library screening,and diagnostic ion filtering was established to systematically screen and characterize dipeptides in C.sinensis.Offline 2D LC integrating hydrophilic interaction LC and reverse phase separations was established to eliminate interference and identify the target dipeptides.A library containing the potential 400 dipeptides was created,and a precursor ion list with all theoretical precursor ions was adopted to trigger the MS/MS scan with high sensitivity.To identify dipeptides,the type and connection sequence of amino acids were determined according to the product ions.Ile and Leu residues were differentiated for the first time according to the characteristic ion at m/z 69.07.Ultimately,170 dipeptides were identified or tentatively characterized from C.sinensis,and most are reported for the first time in this species herein.In addition,the identified dipeptides were also applied for discrimination among the three Cordyceps species,and 11 markers were identified.The obtained results provide a deeper understanding of the chemical basis of C.sinensis.

Pheretima,also called"earthworms",is a well-known animal-derived traditional Chinese medicine that is extensively used in over 50 Chinese patent medicines(CPMs)in Chinese Pharmacopoeia(2020 edi-tion).However,its zoological origin is unclear,both in the herbal market and CPMs.In this study,a strategy for integrating in-house annotated protein databases constructed from close evolutionary relationship-sourced RNA sequencing data from public archival resources and various sequencing al-gorithms(restricted search,open search,and de novo)was developed to characterize the phenotype of natural peptides of three major commercial species of Pheretima,including Pheretima aspergillum(PA),Pheretima vulgaris(PV),and Metaphire magna(MM).We identified 10,477 natural peptides in the PA,7,451 in PV,and 5,896 in MM samples.Five specific signature peptides were screened and then validated using synthetic peptides;these demonstrated robust specificity for the authentication of PA,PV,and MM.Finally,all marker peptides were successfully applied to identify the zoological origins of Brain Heart capsules and Xiaohuoluo pills,revealing the inconsistent Pheretima species used in these CPMs.In conclusion,our integrated strategy could be used for the in-depth characterization of natural peptides of other animal-derived traditional Chinese medicines,especially non-model species with poorly annotated protein databases.

AIM: To observe and compare the clinical efficacy and safety of remimazolam and propofol alone and in combination in endoscopic retrograde cholangiopancreatography (ERCP) anesthesia. METHODS: A total of 120 patients undergoing elective ERCP were divided into the propofol group (P group), the remimazolam group (R group), and the remimazolam combined with propofol group (RP group) according to a random number table, with 40 patients in each group, and the three groups completed anesthesia according to the designated drug regimen (propofol in group P; remimazolam in group R; and remimazolam combined with propofol in group RP). General information, operation time and awakening time of the patients in the three groups were compared, as well as oxygen saturation (SpO