A novel hierarchical machine learning model for hospital-acquired venous thromboembolism risk assessment among multiple-departments.

Venous thromboembolism (VTE) is a common vascular disease and potentially fatal complication during hospitalization, and so the early identification of VTE risk is of significant importance. Compared with traditional scale assessments, machine learning methods provide new opportunities for precise early warning of VTE from clinical medical records. This research aimed to propose a two-stage hierarchical machine learning model for VTE risk prediction in patients from multiple departments. First, we built a machine learning prediction model that covered the entire hospital, based on all cohorts and common risk factors. Then, we took the prediction output of the first stage as an initial assessment score and then built specific models for each department. Over the duration of the study, a total of 9213 inpatients, including 1165 VTE-positive samples, were collected from four departments, which were split into developing and test datasets. The proposed model achieved an AUC of 0.879 in the department of oncology, which outperformed the first-stage model (0.730) and the department model (0.787). This was attributed to the fully usage of both the large sample size at the hospital level and variable abundance at the department level. Experimental results show that our model could effectively improve the prediction of hospital-acquired VTE risk before image diagnosis and provide decision support for further nursing and medical intervention.

Long noncoding RNA DLEU1 aggravates pancreatic ductal adenocarcinoma carcinogenesis via the miR-381/CXCR4 axis.

Recent evidence has highlighted that long noncoding RNAs (lncRNA) are associated with many diseases, particularly cancer. However, current understanding of the lncRNA deleted in lymphocytic leukemia 1 (DLEU1) in pancreatic ductal adenocarcinoma (PDAC) remains limited. Our studies indicated that the DLEU1 expression level was upregulated in PDAC tissue samples compared with adjacent normal tissue. Moreover, the aberrant overexpression of DLEU1 indicated poor prognosis of patients with PDAC. Loss-of-function experiments revealed that DLEU1 knockdown inhibited the proliferation, migration, and invasion of PDAC cells in vitro and decreased tumor growth in vivo. Bioinformatics analysis predicted that miR-381 potentially targeted the DLEU1 3'-untranslated region (UTR), suggesting an interaction between miR-381 and DLEU1. Furthermore, miR-381 also targeted the chemokine receptor-4 (CXCR4) messenger RNA 3'-UTR, which was validated by luciferase reporter assay. Taken together, our study demonstrated the oncogenic role of DLEU1 in clinical PDAC specimens and cellular experiments, showing the potential involvement of DLEU1/miR-381/CXCR4 pathway. These results provide novel insight into PDAC tumorigenesis.

Scutellarin Inhibits Proliferation, Invasion, and Tumorigenicity in Human Breast Cancer Cells by Regulating HIPPO-YAP Signaling Pathway.

BACKGROUND The present study aimed to investigate the effects of Scutellarin on proliferation, invasion and tumorigenicity in human breast carcinoma MCF-7 cells and its associated molecular mechanisms. MATERIAL AND METHODS The MCF-7 cells were cultured with varies of concentrations of Scutellarin in vitro. The proliferation, invasion, and apoptosis of MCF-7 cells were studied via CCK-8 assay, transwell assay, and flow cytometry. In vivo expression of the HIPPO pathway key proteins YAP and p-YAP of MCF-7 cells were analyzed by immunohistochemistry. RESULTS The inhibition rates of Scutellarin-treated MCF-7 cells were 40.1%, 58.7%, and 70.6% for 24, 48, and 72 h, respectively. The MCF-7 cell proliferation was significantly inhibited by Scutellarin. Treating MCF-7 cells with Scutellarin led to invasion inhibition. The rates apoptotic cells were between 12.4±1.9% and 23.9±2.1% in 40-120 µM Scutellarin-administrated groups, which had a significant rise compared with the control group (7.8±1.9%, P<0.05). Scutellarin significantly inhibited MCF-7 xenograft tumor growth. Immunohistochemical analysis showed that the inhibition of tumor growth in Scutellarin-treated mice was associated with increased p-YAP and decreased YAP expression in vivo. CONCLUSIONS Scutellarin-treated breast carcinoma MCF-7 cells had significantly inhibited growth and induced apoptosis, which is associated with induction of autophagy through regulation of the HIPPO-YAP signaling pathway, providing support to the clinical use of Scutellarin-based medication to achieve optimized outcome in patients with breast carcinoma.

Does celecoxib improve the efficacy of chemotherapy for advanced non-small cell lung cancer?

AIMS: Clinical trials have reported conflicting results about whether celecoxib plus chemotherapy improves outcomes over chemotherapy alone in patients with advanced non-small cell lung cancer. METHODS: We performed a meta-analysis comparing the primary and secondary endpoints of treatment with celecoxib plus chemotherapy vs. chemotherapy alone in patients with advanced non-small cell lung cancer. RESULTS: Six eligible trials (1181 patients) were selected from the 206 studies that were identified initially. A significant difference, favouring celecoxib plus chemotherapy over chemotherapy alone, was observed in the overall response rate [odds ratio (OR) 1.34; 95% confidence interval (CI) 1.08, 1.67; P = 0.009). However, there was no difference in the 1-year survival rate (OR 1.08; 95% CI 0.86, 1.35; P = 0.512), clinical benefit (OR 1.05; 95% CI 1.88, 1.25; P = 0.613), complete response (OR 0.77; 95% CI 0.39, 1.51; P = 0.446) or partial response (OR 1.22; 95% CI 0.92, 1.63; P = 0.163). Toxicity did not differ significantly with the exception of the occurrence of leucopenia and thrombocytopenia. CONCLUSIONS: Celecoxib plus chemotherapy appeared to improve the overall response rate compared with chemotherapy alone in the treatment of patients with advanced non-small cell lung cancer. Further prospective randomized controlled trials are now needed.

MicroRNA-17 promotes normal ovarian cancer cells to cancer stem cells development via suppression of the LKB1-p53-p21/WAF1 pathway.

The mechanism underlying the development of human ovarian cancer is poorly understood. The liver kinase protein, LKB1, is hypothesized to play a pivotal role in tumor cell proliferation and invasion capacity through regulation of p53 and p21/WAF1 expression. Previous studies suggest LKB1 may, in turn, be regulated by microRNA-17. Here, we examined the role of miR-17 in the expression of LKB1 and the downstream effects on proliferation and invasion capacity of normal ovarian cancer cells (OCCs) and ovarian stem cells. In this study, both the mRNA and protein expression levels of LKB1, p53, and p21 decreased in OCCs following transfection with a miR-17 expression plasmid. MiR-17 expression affected cell cycle regulation and stimulated the proliferation and invasion capacity of OCCs in vitro. ChIP assays indicated that the binding efficiency of p53 to the p21/WAF1 gene promoter was much lower in miR-17 transfected OCCs than in OCCs transfected with a mutated miR-17. Co-immunoprecipitation and western blotting showed significantly lower levels of p53 and p53 Ser15-pho in the miR-17 transfected OCCs as compared to the mutant miR-17 transfected OCCs. Xenograft experiments confirmed that suppression of tumor growth in vivo occurred in the absence of functional miR-17. These findings suggest that mature miR-17 expression may have an important role in the pathogenesis of human ovarian tumors through its interference with the LKB1-p53-p21/WAF1 pathway expression by epigenetic modification. These findings are of potential importance in the identification of novel therapeutic targets in human ovarian cancer.

EZH2-specific microRNA-98 inhibits human ovarian cancer stem cell proliferation via regulating the pRb-E2F pathway.

The Polycomb group protein, enhancer of zeste homolog 2 (EZH2), plays an important role in transcriptional regulation through chromatin remodeling and interactions with other transcription factors to control cell proliferation and embryonic development. Previous study has shown that EZH2 is important for cell cycle regulation and is highly expressed in human ovarian cancer. Loss of EZH2 inhibits growth of ovarian cancer as well as other human carcinomas. In this study, an expression plasmid of EZH2-targeted microRNA-98 was constructed and transfected into human ovarian cancer stem cells (OCSCs). Seventy-two hours after transfection, cell growth was inhibited and arrested at the G0/G1 transition. p21(CIPI/WAF1) was up-regulated, while the CDK2/cyclin E complex and c-Myc were down-regulated. Most importantly, expression levels of E2F1, retinoblastoma protein (pRb), and histone deacetylase 1 (HDAC1) in the pRb-E2F signaling pathway had changed. Furthermore, microRNA-98 suppressed the growth of OCSCs xenograft tumors. Our findings suggest that EZH2-specific microRNA-98 can effectively inhibit cell proliferation in vitro and regulate the pRb-E2F pathway in human OCSCs.

Pharmacological Validation of ASIC1a as a Druggable Target for Neuroprotection in Cerebral Ischemia Using an Intravenously Available Small Molecule Inhibitor.

Acidosis is a hallmark of ischemic stroke and a promising neuroprotective target for preventing neuronal injury. Previously, genetic manipulations showed that blockade of acid-sensing ion channel 1a (ASIC1a)-mediated acidotoxicity could dramatically alleviate the volume of brain infarct and restore neurological function after cerebral ischemia. However, few pharmacological candidates have been identified to exhibit efficacy on ischemic stroke through inhibition of ASIC1a. In this work, we examined the ability of a toxin-inspired compound 5b (C5b), previously found to effectively inhibit ASIC1a in vitro, to exert protective effects in animal models of ischemic stroke in vivo. We found that C5b exerts significant neuroprotective effects not only in acid-induced neuronal death in vitro but also ischemic brain injury in vivo, suggesting that ASIC1a is a druggable target for therapeutic development. More importantly, C5b is able to cross the blood brain barrier and significantly reduce brain infarct volume when administered intravenously in the ischemic animal model, highlighting its systemic availability for therapies against neurodegeneration due to acidotoxicity. Together, our data demonstrate that C5b is a promising lead compound for neuroprotection through inhibiting ASIC1a, which warrants further translational studies.

Construction of a Risk Prediction Model for Hospital-Acquired Pulmonary Embolism in Hospitalized Patients.

The purpose of this study is to establish a novel pulmonary embolism (PE) risk prediction model based on machine learning (ML) methods and to evaluate the predictive performance of the model and the contribution of variables to the predictive performance. We conducted a retrospective study at the Shanghai Tenth People's Hospital and collected the clinical data of in-patients that received pulmonary computed tomography imaging between January 1, 2014 and December 31, 2018. We trained several ML models, including logistic regression (LR), support vector machine (SVM), random forest (RF), and gradient boosting decision tree (GBDT), compared the models with representative baseline algorithms, and investigated their predictability and feature interpretation. A total of 3619 patients were included in the study. We discovered that the GBDT model demonstrated the best prediction with an area under the curve value of 0.799, whereas those of the RF, LR, and SVM models were 0.791, 0.716, and 0.743, respectively. The sensibilities of the GBDT, LR, RF, and SVM models were 63.9%, 68.1%, 71.5%, and 75%, respectively; the specificities were 81.1%, 66.1, 72.7%, and 65.1%, respectively; and the accuracies were 77.8%, 66.5%, 72.5%, and 67%, respectively. We discovered that the maximum D-dimer level contributed the most to the outcome prediction, followed by the extreme growth rate of the plasma fibrinogen level, in-hospital duration, and extreme growth rate of the D-dimer level. The study demonstrates the superiority of the GBDT model in predicting the risk of PE in hospitalized patients. However, in order to be applied in clinical practice and provide support for clinical decision-making, the predictive performance of the model needs to be prospectively verified.

Hydrogen-rich saline attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels.

OBJECTIVES: The inhaled general anesthetic isoflurane has been shown to induce caspase-3 activation in vitro and in vivo. The underlying mechanisms and functional consequences of this activity remain unclear. Isoflurane can induce caspase-3 activation by causing accumulation of reactive oxygen species (ROS), mitochondrial dysfunction, and reduction in adenosine triphosphate (ATP) levels. This study aimed to investigate the protective effect of hydrogen, a novel antioxidant, against isoflurane-induced caspase-3 activation and cognitive impairment. METHODS: H4 human neuroglioma cells overexpressing human amyloid precursor protein were treated with saline or hydrogen-rich saline (HS, 300 µM), with or without 2% isoflurane, for 6 h or 3 h. Western blot analysis, fluorescence assays, and a mitochondrial swelling assay were used to evaluate caspase-3 activation, levels of ROS and ATP, and mitochondrial function. The effect of the interaction of isoflurane (1.4% for 2 h) and HS (5 mL/kg) on cognitive function in mice was also evaluated using a fear conditioning test. RESULTS: We found that HS attenuated isoflurane-induced caspase-3 activation. Moreover, HS treatment mitigated isoflurane-induced ROS accumulation, opening of mitochondrial permeability transition pores, reduction in mitochondrial membrane potential, and reduction in cellular ATP levels. Finally, HS significantly alleviated isoflurane-induced cognitive impairment in mice. CONCLUSIONS: Our results suggest that HS attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels. These findings warrant further research into the underlying mechanisms of this activity, and indicate that HS has the potential to attenuate anesthesia neurotoxicity.

Role of interleukin-4, the chemokine CCL3 and its receptor CCR5 in neuropathic pain.

Cytokines and chemokines are involved in chronic pain syndromes, and their expression is altered in injury-induced neuronal pain pathways. However, the exact cytokines/chemokines involved and their mechanism of action remain to be determined. In this study, we investigated the role of interleukin-4 and the chemokine/chemokine receptor pair CCL3/CCR5 in a mouse model of chronic constriction injury (CCI)-induced neuropathic pain. Neuropathic pain was induced by surgical ligation of the sciatic nerve and assessed by measuring thermal hyperalgesia and mechanical allodynia using a plantar test and a dynamic plantar esthesiometer. The underlying mechanisms were investigated by real-time quantitative reverse transcription polymerase chain reaction, western blotting, immunohistochemistry, ELISA, and histopathology. CCI-induced neuropathic pain was associated with CCL3 and CCR5 upregulation and microglial activation. Intrathecal injection of the anti-inflammatory cytokine interleukin-4 or CCL3-neutralizing antibody alleviated CCI-induced inflammation, suppressing the CCI-induced upregulation of tumor necrosis factor-α and interleukin-1ß in the serum and spinal cord, restoring the CCI-induced upregulation of CCL3 and CCR5, and suppressing the CCI-induced activation of p38 mitogen-activated protein kinase. Knockout of CCR5 also suppressed CCI-induced neuropathic pain. Since the upregulation of chemokines and cytokines is directly or indirectly involved in chronic pain after nerve injury, these molecules are potential therapeutic targets in the treatment of neuropathic pain.

Isoflurane suppresses the self-renewal of normal mouse neural stem cells in a p53-dependent manner by activating the Lkb1-p53-p21 signalling pathway.

Isoflurane is widely used in anaesthesia for surgical operations. However, whether it elicits unwanted side effects, particularly in neuronal cells, remains to be fully elucidated. The Lkb1-p53-p21 signalling pathway is able to modulate neuronal self­renewal and proliferation. Furthermore, the suppression of Lkb1­dependent p21 induction leads to apoptosis. In the present study, whether Lkb1­p53­p21 signalling is involved in the response to isoflurane was investigated. A comparison of mouse primary, wild­type neural stem cells (WT NSCs) with the p53­/­ NSC cell line, NE­4C, revealed that isoflurane inhibited proliferation in a dose­, a time­ and a p53­dependent manner. However, flow cytometric analysis revealed that the concentration of isoflurane which caused 50% inhibition (the IC50 value) induced cell cycle arrest in WT NSCs. Furthermore, the protein expression levels of LKB1, p53 and p21 were increased, although those of nestin and survivin decreased, following treatment of WT NSCs with isoflurane. On the other hand, isoflurane induced the phosphorylation of Ser15 in p53 in WT NSCs, which was associated with p53 binding to the p21 promoter, and consequentially, the transcriptional activation of p21. All these events were abrogated in NE­4C cells. Taken together, the present study has demonstrated that isoflurane suppresses the self-renewal of normal mouse NSCs by activating the Lkb1-p53-p21 signalling pathway.

Epigenetic silencing of HDAC1 by miR-449a upregulates Runx2 and promotes osteoblast differentiation.

Human-induced pluripotent (iPS) cells can be induced to differentiate into osteoblasts, but the process is inefficient and time-consuming. Previous studies indicated a close association between the expression of Runx2 and osteoblast differentiation, and established that the transcriptional activation of the Runx2 gene was closely associated with histone acetylation. microRNA-449a (miR-449a) represses HDAC1 expression, thereby regulating histone acetylation. In the present study, whether the expression of miR-449a enhanced the generation of osteoblasts from human iPS cells was investigated. Introduction of miR-449a into human iPS cells resulted in the expression of osteoblast markers after only four days, compared to eight days for untransfected human iPS cells. Differentiation to osteoblasts was associated with a reduction in HDAC1 expression, and higher levels of histone acetylation, particularly at the binding sites on the Runx2 promoter in the human miR-449a-transfected iPS cells. Silencing of endogenous HDAC1 expression by exogenous miR-449a therefore maintains histone acetylation status, stimulates Runx2 gene expression and rapidly promotes osteoblast differentiation.

The induction of rat spermatogonial stem cells into neuronal-like cells and behavioral recovery following transplantation in a rat Parkinson's disease model.

Parkinson's disease (PD) is a widespread age-associated neurodegenerative disorder. Current treatment is symptomatic rather than curative. However, stem cell replacement therapies may have the potential to offer curative treatment. In this study, we demonstrate that rat CD49f+ spermatogonial stem cells (rSSCs) can be induced to become functional dopaminergic neuron-like cells in vitro. Furthermore, when rSSCs were transplanted into 6-hydroxydopamine (6-OHDA)-treated PD rats, the results indicated that rSSCs expressed multiple neuron cell markers and were ameliorative to behavioral recovery in PD rats after induction both in vitro and in vivo. In addition, rSSCs demonstrated increased activity in the regeneration of dopaminergic neuron-like cells, increased migration distances and were associated with improvement in animal behavior in the PD rat model. Therefore, rSSCs could be a source of dopaminergic neuron-like cells with potential benefit in cell replacement therapy for PD.