1.Reduced apoptosis induced by endotoxin in mice blocking TRAIL with soluble death receptor 5
Huiling BAI ; Xueyin WANG ; Shulian LI ; Hongying HUANG ; Yaowu DU ; Guangchao LIU ; Yuanfang MA
Chinese Journal of Microbiology and Immunology 2009;29(2):151-155
Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.
2.Study on differentiation of bone marrow mesenchymal stem cells into neurons induced by bone morphogenetic protein 7 in vitro
Guangchao BAI ; Wen ZHANG ; Lei GAO ; Kuanxin LI
Chinese Journal of Orthopaedics 2020;40(12):802-810
Objective:To explore the role of BMP7 in inducing the differentiation of BMSCs into neurons in vitro.Methods:BMSCs were isolated and cultured by whole bone marrow adherence method. Adipogenic induction and osteogenic differentiation were used to test the multi-directional differentiation ability of BMSCs. BMSCs were randomly divided into control group and BMP7 groups (25 ng/ml, 50 ng/ml, 75 ng/ml and 100 ng/ml). The effect of BMP7 on the proliferation of rat BMSCs was measured by MTT assay. BMP7 induced morphological changes in rat BMSCS under an inverted phase contrast microscope. The relative expression levels of NF200, SYN1, MAP2 and GFAP mRNA in induced cells were measured by qRT-PCR. Immunofluorescence was used to measure the expression of NSE protein.Results:Rat BMSCs showed lipid droplets in the cytoplasm after adipogenic induction, and oil red O staining was positive; rat BMSCs showed opaque mineral nodules after osteogenic induction, and alizarin red stained was positive. At 2 d, 3 d, 4 d, 5 d, and 6 d after induction, the cell absorbance values of each group were statistically different ( P < 0.05). On the 6th day of induction, the absorbance values of the control group, 25 ng/ml BMP7 group, 50 ng/ml BMP7 group, 75 ng/ml BMP7 group and 100 ng/ml BMP7 group were 0.370±0.003, 0.399±0.003, 0.404±0.003, 0.410±0.003, 0.397±0.001, respectively. Cells absorbance value of 75 ng/ml BMP7 group was significantly higher than the other groups ( P< 0.05). The 75 ng/ml BMP7 group had the most significant changes in cell morphology, similar to neurons in cell morphology. The relative expression of NF200, SYN1, MAP2, and GFAP mRNA of 75 ng/ml BMP7 group (5.47±0.59, 1.48±0.38, 2.86±1.65, 4.41±0.13) was significantly higher than that of the control group ( P< 0.05). The positive rate of NSE immunofluorescence staining in the 75 ng/ml BMP7 group was higher than that in the control group (32.94%±1.62% vs 0). Conclusion:BMP7 has the ability to induce the differentiation of rat BMSCs into neurons in vitro.
3.A cross-sectional study of early-onset epilepsy of intracerebral hemorrhage and construction of a risk prediction model
Xiangyan BAI ; Liang ZHANG ; Hailin LI ; Dengjun GUO ; Guangchao YIN
Chinese Critical Care Medicine 2022;34(12):1273-1279
Objective:To study the early-onset epilepsy of intracerebral hemorrhage and build a prediction model to evaluate its prediction efficiency.Methods:A cross-sectional investigation was conducted to construct a specialized optimized prediction model. The prediction model was converted into a visual optimized scoring scale, so as to quantify the probability of secondary epilepsy after intracerebral hemorrhage. Based on the current prediction model of acute cerebral infraction and post-stroke seizure (AIS-PSS), the evaluation efficacy of optimized score for secondary epilepsy after hemorrhagic stroke was explored.Results:① After sample size calculation and sufficient inclusion and exclusion, 159 patients with cerebral hemorrhage were continuously selected as the model group of this cross-sectional study. A total of 29 patients with early-onset epilepsy and 130 patients without secondary epilepsy were enrolled. The time span was from January 2021 to August 2021. In addition, 77 patients with acute cerebral hemorrhage from August 2021 to February 2022 were selected as the verification group, among which 12 patients had early-onset epilepsy and 65 patients had not any secondary epilepsy. ② There were significant differences in demographic characteristics such as diabetes history, cerebral infarction history, smoking history, National Institutes of Health Stroke Scale (NIHSS) score, intracerebral hemorrhage hematoma volume, serum creatinine (SCr), neuron-specific enolase (NSE), S-100 protein and intracerebral hemorrhage site between the two model groups with different prognosis (all P < 0.05). ③ The above indexes were included in univariate and multivariate Poisson regression analysis, and the results showed that the duration of diabetes [relative risk ( RR) = 1.229, 95% confidence interval (95% CI) was 1.065-1.896, P = 0.036], smoking history ( RR = 1.419, 95% CI was 1.133-2.160, P = 0.030), history of cerebral infarction ( RR = 1.634, 95% CI was 1.128-2.548, P = 0.041), hematoma volume of cerebral hemorrhage ( RR = 1.222, 95% CI was 1.024-2.052, P = 0.041), NES content ( RR = 1.146, 95% CI was 1.041-1.704, P = 0.032), were independent influencing factors to constitute the prediction model. The prediction model was converted into a visual optimized scoring scale in the form of a line diagram to obtain the prediction probability corresponding to the corresponding score. ④ Receiver operator characteristic curve (ROC curve) was used to test the evaluation efficiency of optimized score and AIS-PSS score for early-onset cerebral hemorrhage epilepsy. Relevant data of patients in the verification group were extracted according to the information of two scores, and the final score of each patient in the verification group was obtained. The score and prognosis were put into the ROC curve to evaluate the predictive ability of different prediction models. The results showed that the cut-off value of the optimized score and the AIS-PSS score were 144 points and 7 points, respectively, and the area under the ROC curve (AUC) and the Yoden index of the optimized score were slightly lower than the AIS-PSS score. However, compared with AIS-PSS score, there was no significant difference in the evaluation efficiency of optimized score for early-onset epilepsy ( Z = 1.874, P > 0.05). Conclusion:This study constructed a specific early-onset epilepsy prediction model for patients with hemorrhagic stroke, and transformed it into an optimized score that is easy for clinical use, and its evaluation efficiency is reliable.