Objective To explore the effect of Ulinastatin on blood brain barrier (BBB) and apoptosis of neural cells in septic rats.Methods Fifty-two clean level male Sprague-Dawley rats were randomly (random number table) divided into six groups:Sham groups at 6 h and 24 h,each group with six rats.Sepsis groups (CLP) and Ulinastatin treated groups (UTI) at 6 h and 24 h,each group with ten rats.In CLP and UTI groups,cecal ligation and puncture (CLP) were performed to induce sepsis.Sham group was only opened and closed abdomen.Ulinastatin (50 000 U/kg) was administered via femoral vein 1 h after CLP.The same volume of saline instead of Ulinastatin was administered in Sham and CLP groups.The neurological status was assessed by Neurological Deficit Scale Scores (NDSS) at 6 h and 24 h after CLP.Then the brain was harvested for HE staining and weighing water content.The BBB permeability was assayed by Evans Blue dye extravasations.Apoptosis of neural cells were detected by TUNEL immune fluorescence.Statistical analysis was performed with SPSS version 13.0,ANOVA was used for multiple groups comparison and t-test for paired comparison.Results The Neurological Deficit Scale Scores of UTI group was lower than Sham group (P ＜ 0.05) but higher than that of CLP group (P ＜ 0.05).Swelling,degeneration and edema were observed in cerebral cortex and hippocampal neurons in CLP group through light microscope,and were more serious than those in UTI group.Compared with UTI 24 h group,BBB permeability of CLP 24 h group significantly rose (P ＜ 0.05).The number of apoptosis of neural cells increased more in CLP group than it did in UTI group (P ＜ 0.05).Conclusions Ulinastatin could protect the cerebral tissue in septic rats by alleviating the damage of BBB and reducing the apoptosis of neural cells.

Objective To explore the effect of autophagy regulator on the injury of rat hippocampal neurons induced by oxygen-glucose deprivation (OGD).Methods Rat hippocampal neurons were cultivated in primary and subjected to OGD to simulate neuronal hypoxic ischemia injury for 2 hours or 6 hours followed by reperfusion for 12 hours with or without 3-methyladenine (3-MA, 20μmol/L) or rapamycin (0.2μmol/L). The morphology of neurons was observed with optical microscope. The expression of autophagy-related protein (LC3, P62) and apoptosis-related protein (cleaved caspase-3) were assessed by Western Blot analysis. The apoptosis of neurons was detected by flow cytometry, the release rate of lactate dehydrogenase (LDH) was calculated by automatic biochemical analyzer, and the cell activity was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay.Results Compared with the control group, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.091±0.160, 3.422±0.186 vs. 0.256±0.021), cleaved caspase-3 (gray value: 0.230±0.025, 0.440±0.051 vs. 0.050±0.007), neuronal apoptotic rate, LDH release rate [(38.50±4.15)%, (59.60±5.65)% vs. (12.40±1.32)%] were increased, while the expression of P62 (gray value: 0.290±0.025,0.120±0.026 vs. 0.450±0.040), neuronal activity [(71.40±7.23)%, (42.80±4.12)% vs. (100.30±2.30)%] were decreased at 2 hours or 6 hours after OGD (allP < 0.05). When the time of OGD was 2 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ (gray value: 2.281±0.121), the neuronal activity [(51.10±5.73)%] were decreased, while the expression of P62 and cleaved caspase-3 (gray scale: 0.410±0.037, 0.330±0.027, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (47.30±4.43)%] were increased (allP < 0.05). When the time of OGD was 2 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.689±0.214), the neuronal activity [(85.30±8.56)%] were increased, while the expression of P62 and cleaved caspase-3 (gray value: 0.170±0.040, 0.090±0.096, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (24.30±2.14)%] were decreased (allP < 0.05). On the contrary, when the time of OGD was 6 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.021±0.178, 0.240±0.017), neuronal apoptotic rate, the injury of neurons [LDH release rate: (36.60±3.45)%] were decreased, while the expression of P62 (gray value: 0.350±0.060), the neuronal activity [(59.70±6.13)%] were increased (allP < 0.05). When the time of OGD was 6 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.923±0.201, 0.590±0.062), neuronal apoptotic rate, the injury of neurons [LDH release rate:(71.20±7.81)%] were increased, while the expression of P62 (gray value: 0.070±0.008), the neuronal activity [(27.30±2.12)%] were decreased (allP < 0.05).Conclusion The enhancement of autophagy has protective effect on neurons under the condition of mild OGD, while it can aggravate the injury of neurons induced by a long-time OGD.

Objective To investigate immunological dysfunction of intestine mucosa barrier in a rat model of sepsis. Method Sixty Sprague-Dawley rats were assigned randomly(random number) into sepsis group (n = 45)and control group (n = 15). The animals in sepsis group were subjected to cecal ligation and puncture (CLP), whereas rats of control group underwent a sham surgery. The ileac mucosa and segments were harvested 3 h, 6 h and 12 hours after CLP, and the blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3(TFF_3) mRNA, lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected. Results In the septic animals, in-testinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of the lamina propria, hemorrhage and ulceration. Compared with control, the expression of TFF_3 mRNA and level of RD-5 pro-tein were decreased and the mucosal lymphocyte apoptosis increased (P < 0.05) in sepsis group. Compared with control group, the significant differences in RD-5 and TFF_3 mRNA appeared 3 hours after CLP and those differ-ences were progressively increased in 6 hours and 12 hours after CLP in sepsis group (P < 0.05, F of RD-5 = 11. 76, F of TFF_3 = 16.86 and F of apoptosis = 122.52). In addition, the gut-origin bacterial DNA in plasma de-tected was positive in all sepsis animals. Conclusions It suggests that immunological function of intestinal mucosa is impaired in septic rats and further worsened following the course of sepsis.

The sensitivity of cardiac troponin testing for diagnosing acute myocardial infarction(AMI)varies over time from chest pain onset.This study aimed to determine the diagnostic performance of point-of-care testing cardiac troponin T(POCT-cTnT)at different time intervals post-symptom onset to refine rapid rule-out approaches for AMI.Methods This retrospective study included 6,024 patients presenting with chest pain from January 2018 to December 2022.POCT-cTnT and central lab cTnI levels were measured on admission.Receiver operating charac-teristics analysis stratified by time windows assessed the accuracy of POCT-cTnT for diagnosing AMI.Results The overall AUC of POCT-cTnT for diagnosing AMI was 0.826(95%CI:0.816～0.836),with a sensitivity of 72.81%and a specificity of 86.26%.According to the time intervals of chest pain onset(＜3 hours,3～6 hours,6～12 hours,12～24 hours,24～72 hours,and≥72 hours),the AUC values for groups after 6～12 hours were 0.918,0.928,0.920 and 0.908,respectively,with no statistically significant difference(P＞0.05),but all were higher than the groups within 6 hours(P＜0.001).According to the time of chest pain onset,the AUC for the≥8h group was 0.921,with a negative predictive value(NPV)of 98.1%and a negative likelihood ratio(-LR)of 0.11.Its AUC was higher than those of the≥3 h,≥2 h,≥1 h,and overall groups(P＜0.05),but there was no statistically significant differ-ence compared with the groups after≥4 h(P＞0.05).Conclusions Chest pain onset time has a certain impact on the performance of a single POCT-cTnT test for diagnosing AMI.The duration from chest pain onset to hospital admission combined with POCT-cTnT test may improve the reliability in diagnosing AMI.Specifically,a single POCT-cTnT test at four hours after chest pain onset,especially eight hours after chest pain onset,can diagnose or exclude AMI.