In recent years, epileptic seizure detection based on electroencephalogram (EEG) has attracted the widespread attention of the academic. However, it is difficult to collect data from epileptic seizure, and it is easy to cause over fitting phenomenon under the condition of few training data. In order to solve this problem, this paper took the CHB-MIT epilepsy EEG dataset from Boston Children's Hospital as the research object, and applied wavelet transform for data augmentation by setting different wavelet transform scale factors. In addition, by combining deep learning, ensemble learning, transfer learning and other methods, an epilepsy detection method with high accuracy for specific epilepsy patients was proposed under the condition of insufficient learning samples. In test, the wavelet transform scale factors 2, 4 and 8 were set for experimental comparison and verification. When the wavelet scale factor was 8, the average accuracy, average sensitivity and average specificity was 95.47%, 93.89% and 96.48%, respectively. Through comparative experiments with recent relevant literatures, the advantages of the proposed method were verified. Our results might provide reference for the clinical application of epilepsy detection.
To isolate and culture adi pose-derived stem cells (ADSCs), and to study the effects of the conditioned medium of ADSCs (ADSC-CM) treated with insul in on HaCaT cells. Methods ADSCs were isolated from adipose tissue donated by the patient receiving abdominal surgery and were cultured. The concentration of ADSCs at passage 3 was adjusted to 5 × 104 cells/mL. The cells were divided into 2 groups: group A in which the cells were incubated in 1 × 10-7 mol/ Linsul in for 3 days, and group B in which the cells were not treated with insul in. ADSC-CM in each group was collected 3 days after culture, then levels of VEGF and hepatocyte growth factor (HGF). HaCaT cells were cultured and the cells at passage 4 were divided into 4 groups: group A1, 0.5 mL 2% FBS and 0.5 mL ADSC-CM from group A; group B1, 0.5 mL 2% FBS and 0.5 mL ADSC-CM from group B; group C1, 1 mL 2% FBS of 1 × 10-7 mol/ L insul in; group D1, 1 mL 2%FBS. Prol iferation of HaCaT cells was detected by MTT method 3 days after culture, apoptosis rate of HaCaT cells was measured by Annexin V-FITC double staining 12 hours after culture, and the migration abil ity was measured by in vitro wound-heal ing assay 0, 12, 24, 36 and 48 hours after culture. Results The level of VEGF in groups A and B was (643.28 ± 63.57) and (286.52 ± 46.68) pg/mL, respectively, and the level of HGF in groups A and B was (929.95 ± 67.52) and (576.61 ± 84.29) pg/mL, respectively, suggesting differences were significant between two groups (Plt; 0.05). Cell prol iferation detection showed the absorbance value of HaCaT cells in group A1, B1, C1 and D1 was 0.881 ± 0.039, 0.804 ± 0.041, 0.663 ± 0.027 and 0.652 ± 0.042, respectively, suggesting there was significant difference between groups A1 and B1 and groups C1 and D1 (P lt; 0.01), group A1 was significantly higher than group B1 (P lt; 0.05). The apoptosis rate of HaCaT cells in groups A1, B1, C1 and D1 was 5.23% ± 1.98%, 8.82% ± 2.59%, 31.70% ± 8.85% and 29.60% ± 8.41%, respectively, indicating there was significant difference between groups A1 and B1 and groups C1 and D1 (P lt; 0.05), group B1 was significantly higher than group A1 (P lt; 0.05). The migration distance of HaCaT cells in groups A1, B1,C1 and D1 at 36 hours was (0.184 6 ± 0.019 2), (0.159 8 ± 0.029 4), (0.059 2 ± 0.017 6) and (0.058 2 ± 0.012 3) mm, respectively, whereas at 48 hours, it was (0.231 8 ± 0.174 0), (0.205 1 ± 0.012 1), (0.079 2 ± 0.008 1) and (0.078 4 ± 0.011 7) mm, respectively, suggesting there were significant differences between groups A1 and B1 and groups C1 and D1 at 36 and 48 hours (P lt; 0.01), group A1 was significantly higher than group B1 (P lt; 0.05) at 36 and 48 hours, no significant difference was evident at other time points(P gt; 0.05). Conclusion ADSCs treated with insul in can significantly promote the prol iferation and the migration of HaCaT cells and inhibit their apoptosis.
Aiming at the problem that the small samples of critical disease in clinic may lead to prognostic models with poor performance of overfitting, large prediction error and instability, the long short-term memory transferring algorithm (transLSTM) was proposed. Based on the idea of transfer learning, the algorithm leverages the correlation between diseases to transfer information of different disease prognostic models, constructs the effictive model of target disease of small samples with the aid of large data of related diseases, hence improves the prediction performance and reduces the requirement for target training sample quantity. The transLSTM algorithm firstly uses the related disease samples to pretrain partial model parameters, and then further adjusts the whole network with the target training samples. The testing results on MIMIC-Ⅲ database showed that compared with traditional LSTM classification algorithm, the transLSTM algorithm had 0.02-0.07 higher AUROC and 0.05-0.14 larger AUPRC, while its number of training iterations was only 39%-64% of the traditional algorithm. The results of application on sepsis revealed that the transLSTM model of only 100 training samples had comparable mortality prediction performance to the traditional model of 250 training samples. In small sample situations, the transLSTM algorithm has significant advantages with higher prediciton accuracy and faster training speed. It realizes the application of transfer learning in the prognostic model of critical disease with small samples.
Objective To investigate the effect of microRNA-22-3p (miR-22-3p) on the inflammation of human pulmonary microvascular endothelial cells (HPMEC) induced by lipopolysaccharide (LPS) by regulating the HMGB1/NLRP3 pathway. Methods miRNA microarray was taken from peripheral blood of patients with acute respiratory distress syndrome (ARDS) caused by abdominal infection and healthy controls for analysis, and the target miRNA was selected. miRNA mimics, inhibitor and their negative controls were transfected in HPMECs which were stimulated with LPS. Real time fluorescent quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the mRNA and protein levels of high mobility group box-1 protein (HMGB1) and nucleotide binding oligomerization segment like receptor family 3 (NLRP3). RT-qPCR and enzyme linked immunosorbent assay were used to detect the levels of inflammatory factors in the cells and supernatant. Results miRNA microarray showed that miR-22-3p was down-regulated in the plasma of patients with ARDS. Compared with the negative control group, after miR-22-3p over-expression, the protein and mRNA levels of HMGB1 and NLRP3 decreased significantly. Similarly, the level of cleaved-caspase-1 decreased significantly. At the same time, interleukin (IL)-6, IL-8 and IL-1β mRNA level in cytoplasm and supernatant were down-regulated by miR-22-3p mimics. After transfected with miR-22-3p inhibitor, the expression levels of HMGB1, NLRP3, caspase-1 protein and inflammatory factors were significantly up-regulated. Conclusion miR-22-3p is significantly downregulated in peripheral blood of ARDS patients caused by abdominal infection, which can inhibit the expression of HMGB1 and NLRP3 and its downstream inflammatory response in HPMECs.
Objective To observe the protective effects of unfractionated heparin (UFH) on high-mobility group box-1 protein (HMGB1) induced increased permeability of endothelial cells, and investigate the protective mechanism of UFH on HMGB1 induced defective expression of zonula occludens-1 (ZO-1). Methods Human umbilical vascular endothelial cells (HUVECs) were culturedin vitro and divided into 4 groups (n=5), namely a control group, a HMGB1 group (100 ng/ml), a heparin group (UFH 10 U/ml), a HMGB1/heparin group (100 ng/ml HMGB1 + UFH 10 U/ml). Endothelial cell viability was measured by methyl thiazolyl tetrazolium (MTT) colorimetric method. Endothelial permeability was determination by Transwell chamber method. Immunofluorescence and laser confocal microscopy were used to assess the distribution of ZO-1. The protein expressions of tight junction protein ZO-1 and nuclear factor kappa B (NF-κB) were detected by Western blot. Results HMGB1 (100 ng/ml) had no inhibitory effect on endothelial cell viability (P＞0.05). UFH pretreatment could reduce the permeability increment of endothelial cells induced by HMGB1. UFH pretreatment could reduce the close loop reduction and damage of ZO-1 induced by HMGB1, enhance the fluorescence intensity and expression of ZO-1, and decrease the NF-κB translocation. Conclusions UFH can protect HMGB1-mediated defect of ZO-1 expression and increased permeability of the endothelial cells. The mechanism may be related to the decreased nuclear translocation of NF-κB.
ObjectiveTo investigate the effect and mechanism of ulinastatin to ventilator induced lung injury (VILI). MethodsTotal 24 SD rats were randomly divided into a control group, a VILI group, and a VILI+ ulinastatin group. High mobility group box-1 (HMGB-1), tumor necrosis factor (TNF)-α and interleukin (IL)-6 in bronchoalveolar lavage fluid, toll like receptor-4, dry/wet ratio and pathological scores of lung tissue were detected in the three groups. ResultsHMGB-1, TNF-α, and IL-6 in bronchoalveolar lavage fluid, toll like receptor-4, dry/wet ratio and pathological scores of lung tissue were significantly higher in the VILI group than those in the control group with statistical differences (P<0.05). While HMGB-1, TNF-α, and IL-6 in bronchoalveolar lavage fluid, toll like receptor-4, dry/wet ratio and pathological scores of lung tissue were reduced in the VILI+ ulinastatin group compared with those in the VILI group. ConclusionUlinastatin may protect ventilator induced lung injury by reducing inflammation level in lung through HMGB-1-TLR4 pathway.
ObjectiveTo investigate the effect of Wnt5a derived from tumor-associated fibroblasts （CAFs） on the migration and invasion of gastric cancer cells. MethodsThe differentially expressed genes Wnt5a between CAFs and normal gastric fibroblasts (NGFs) in gastric cancer tissues and their corresponding normal gastric tissues using the GEO database GSE194261 dataset were screened. Immunohistochemical method was used to detect the expression of Wnt5a protein in tissue samples of clinical gastric cancer patients, and the relationship between Wnt5a protein expression and clinicopathological features of gastric cancer was analyzed. CAFs and NGFs were extracted from fresh surgical specimens of gastric cancer patients, and the expression of Wnt5a in CAFs was detected by real-time fluorescence quantitative-polymerase chain reaction and Western blot experiment. Transwell invasion and migration experiment was used to observe the effects of CAFs, inhibition of Wnt5a expression in CAFs and different concentrations of recombinant Wnt5a protein on the migration and invasion ability of gastric cancer MGC-803 and MKN-28 cell lines in vitro. ResultsThrough the screening of GEO database GSE194261 data set, it was found that Wnt5a was more expressed in CAFs than NGFs (P<0.05). Immunohistochemical results showed that the expression of Wnt5a protein in gastric cancer tissues was significantly stronger than that in normal gastric tissues (P<0.05), and the expression of Wnt5a protein was related to T stage of tumor (χ2=5.035, P<0.05), but not related to gender, age, degree of tumor differentiation, lymph node metastasis, vascular invasion and nerve invasion (P>0.05). Inhibiting Wnt5a derived from CAFs could inhibit the invasion and migration of gastric cancer cells. By stimulating gastric cancer cells with different concentrations of human recombinant Wnt5a protein, it was found that when the concentration of human recombinant Wnt5a protein was greater than 100 ng/mL, the invasion and migration abilities of MGC-803 and MKN-28 gastric cancer cells were significantly increased (P<0.05). ConclusionWnt5a is highly expressed in CAFs derived from the interstitial tissue of gastric cancer, which is related to the invasion depth of gastric cancer and can promote the invasion and migration of gastric cancer cells.
ObjectiveTo investigate the influence of hypoxia on pro-metastasis induced by epithelial-mesenchymal transition (EMT) of human lung adenocarcinoma. MethodsThe human lung cancer cell line H460 was cultured in hypoxic condition and the morphologic changes of the cells were observed under the microscope. The EMT-related markers including E-cadherin and vimentin were detected by Western blot. Transwell migration assay and transwell invasion assay were employed to detect the migratory and invasive activity of cancer cells. ResultsHypoxic induced morphological changes were consistent with the mesenchymal phenotype, such as an elongated fibroblastic morphology, and conversion from a tightly packed epithelial cobblestone pattern to a loosely packed scattered phenotype. Compared with the control group, hypoxic attenuated the quantity of E-cadhenrin, but increased vimentin, which resulted in promotion of migration and invasion of H460. ConclusionHypoxia induces EMT in H460 and enhances the invasive and migratory abilities of lung cancer cells.
The article aims to explore the optimal concentration of arsenic trioxide (As2O3) on HepG2 of liver cancer cells, and the effect of As2O3 on the migration, invasion and apoptosis of HepG2 cells. In this study, the activity of HepG2 cells treated with 0, 1, 2, 4, 8, 16, 32 μmol/L As2O3 was tested by CCK-8 method, the semi-inhibitory concentration (IC50) was calculated, and the morphological changes of HepG2 cells were observed after the action of As2O3 at IC50 concentration for 12, 24, 48 h. The effect of As2O3 on cell migration and invasion ability was verified by wound healing experiment and Transwell invasion experiment. Western blot and qRT-PCR were used to detect the effects of As2O3 on the gene and protein expression levels related to cell migration, invasion and apoptosis. The results showed that, compared with the control group, the activity of HepG2 cells decreased with the increase of the concentration of As2O3 treatment, showing a dose-dependent effect, and its IC50 was 7.3 μmol/L. After 24 hours’ treatment with 8 μmol/L As2O3, HepG2 cells underwent significant apoptosis, and its migration and invasion abilities were significantly reduced. In addition, the protein expression levels of RhoA, Cdc42, Rac1 and matrix metalloproteinase-9 (MMP-9) were down-regulated, the protein and mRNA expression levels of anti-apoptotic gene Bcl-2 were significantly down-regulated, and the protein and mRNA expression levels of pro-apoptotic genes Bax and Caspase-3 were significantly up-regulated. The above results indicate that certain concentration of As2O3 can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.
Objective To explore the effects of DNA cross-linking repair 1B (DCLRE1B) gene on the migration and invasion ability of hepatocellular carcinoma cell. Methods Bioinformatics analysis was used to analyze the expression of DCLRE1B mRNA in hepatocellular carcinoma, and its relationship with the prognosis and related influencing factors of patients. Immunohistochemical staining was used to detect the expression of DCLRE1B protein in resected hepatocellular carcinoma tissues and their corresponding normal liver tissues. The DCLRE1B gene silenced Huh7 and HepG2 hepatocellular carcinoma cell lines were constructed by lentivirus, and the transfected effect was detected by Western blot. The migration and invasion of DCLRE1B silenced hepatocellular carcinoma cells were detected by scratch test and Transwell method. The changes of genes related to epithelial mesenchymal transformation (EMT) after DCLRE1B silencing were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results ① The biological information analysis results showed that: The mRNA expression of DCLRE1B was highly expressed in a variety of tumors including hepatocellular carcinoma (P<0.05). The mRNA expression of DCLRE1B was associated with the TNM staging of tumor (P<0.05). The relative expression level of DCLRE1B mRNA in hepatocellular carcinoma patients was related to their prognosis. The overall survival situation (P=0.038) and progression free survival situation (P=0.005) of hepatocellular carcinoma patients in the high expression group were worse than those in the low expression group. Univariate and multivariate Cox analysis showed that the expression of DCLRE1B gene was an independent factor affecting the prognosis of hepatocellular carcinoma (P<0.05). ② The positive rate of DCLRE1B protein expression in resected hepatocellular carcinoma tissues was higher than that in normal liver tissues (P<0.05). ③ Cell experiment results showed that: After stable silencing DCLRE1B gene of hepatocellular carcinoma cell (Huh7 and HepG2) constructed by lentivirus, the expression of DCLRE1B protein was significantly down regulated (P<0.05). After silencing DCLRE1B gene, the migration and invasion ability of hepatocellular carcinoma cells were significantly decreased (P<0.05). After silencing DCLRE1B, the mRNA expressions of E-cadherin, matrix metalloproteinase 9, and β-catenin were up regulated (P<0.05), and the mRNA expressions of N-cadherin and Vimentin were down regulated (P<0.05), but the mRNA expression of zinc finger transcription factor had no significant change, and the difference was not statistically significant (P＞0.05). Conclusion Silencing DCLRE1B gene can inhibit the migration and invasion ability of hepatocellular carcinoma cells, and its mechanism may be related to the process of EMT.