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The egg production rate is a crucial metric in animal breeding, subject to biological and environmental influences and exhibits characteristics of small sample sizes and non-linearity. Currently, egg production rate prediction research predominantly focuses on single-step point prediction, lacking multistep and interval prediction exploration. To bridge these gaps, this study proposes a recursive, multistep interval prediction method for egg production rates, integrating environmental variables and attention mechanisms. Initially, this study employed three gradient boosting tree models (XGBoost, LightGBM, CatBoost) and the recursive feature elimination (RFE) method to select critical environmental variables and reduce data dimensionality. Subsequently, by scaling the time scale of important environmental variables and utilizing the variational modal decomposition improved by the grey wolf optimization (GWO-VMD) method for time-series decomposition, the volume of important environmental variable data is augmented and its complexity is reduced. Applying the long short-term memory (LSTM) neural network to obtain direct multistep predictions on IMFs, the predicted outcomes are averaged daily to yield the environmental variables for the upcoming two days. Finally, a multistep interval prediction model based on Seq2seq-Attention and Gaussian distribution is proposed in this study, and parameter optimization is carried out using the multi-objective grey wolf optimization algorithm (MOGWO). By inputting the historical egg production rate data and environmental variables into the proposed model, it is possible to achieve multistep point and interval prediction of egg production rates. This method was applied to analyze a dataset of egg production rates of waterfowl. The study demonstrated the feasibility of the recursive multistep prediction approach combined with environmental variables and guides egg production estimation and environmental regulation in animal husbandry.

Background:Lung cancer is one of the most common causes of cancer-related death worldwide due to its rapid growth and metastasis. TIGAR plays a role in promoting survival in various cancer cells, but the molecular mechanisms on metabolic reprogram in tumor cells are not fully understood. Methods: TIGAR expression was detected in lung cancer by western blot and immunohistochemistry. A shRNA interference system was used to knockdown the TIGAR in NSCLC cell lines to delineate its role in NSCLC tumor proliferation using in vitro functional assays and in vivo mouse models. Finally, transcriptomics and metabolomics were used to identify the mechanism of TIGAR. Results: The expression of TIGAR in tumor tissues of NSCLC patients was significantly higher than that of adjacent tissues, which was associated with poor prognosis of NSCLC. The proliferation of NSCLC cells in vitro and the growth of xenografted tumors in vivo were significantly inhibited by TIGAR knockdown. With a combination of transcriptomics and metabolomics, we found that TIGAR maintained intracellular deoxyribonucleotide levels by regulating the expression of the ribonucleotide reductase (RNR) subunit RRM2B, a protein involved in deoxynucleotide synthesis. Further studies with LC-MS and co-immunoprecipitation revealed that TIGAR interacted with the transcription factor YBX1, participated in its phosphorylation and nuclear translocation to induce the expression of its downstream gene RRM2B. In addition, overexpression of RRM2B or exogenous supplementation of dNTPs effectively rescued the restriction of cell proliferation and DNA repair caused by TIGAR knockdown. Conclusion: The present studies revealed a novel mechanism of TIGAR in promoting the proliferation and DNA repair of NSCLC cells through maintaining the intracellular deoxynucleotide level via the YBX1-RRM2B axis.

Amyloid-β peptide (Aβ) aggregation is the hallmark of Alzheimer’s disease (AD). The imbalance between the production and clearance of Aβ results in the accumulation and aggregation of Aβ in the brain. Thus far, few drugs are available for AD treatment, but exercise has been recognized for its cognition-enhancing properties in AD patients. The underlying mechanisms remain unclear. Our recent study showed that long-term running exercise could activate the lysosomal function in the brains of mice. In this study, we investigated whether exercise could reduce Aβ accumulation by activating lysosomal function in APP/PSEN1 transgenic mice. Started at the age of 5 months, the mice were trained with a running wheel at the speed of 18 r/min, 40 min/d, 6 d/week for 5 months, and were killed at the end of the 10th month, then brain tissue was collected for biochemical analyses. The cognitive ability was assessed in the 9th month. We showed that long-term exercise significantly mitigated cognitive dysfunction in AD mice, accompanied by the enhanced lysosomal function and the clearance of Aβ in the brain. Exercise significantly promoted the nuclear translocation of transcription factor EB (TFEB), and increased the interaction between nuclear TFEB with AMPK-mediated acetyl-CoA synthetase 2, thus enhancing transcription of the genes associated with the biogenesis of lysosomes. Exercise also raised the levels of mature cathepsin D and cathepsin L, suggesting that more Aβ peptides could be degraded in the activated lysosomes. This study demonstrates that exercise may improve the cognitive dysfunction of AD by enhancing lysosomal function.

Although early esophageal squamous cell carcinoma (EESCC) with cirrhosis is a relatively rare clinical phenomenon, the management of EESCC in cirrhotic patients continues to be a challenge.To evaluate the feasibility, safety, efficacy and long-term survival outcomes of endoscopic submucosal tunnel dissection (ESTD) for treating EESCC in patients with cirrhosis.This was a single-center retrospective cohort study. We examined 590 EESCC patients who underwent ESTD between July 14, 2014, and May 26, 2021, from a large-scale tertiary hospital. After excluding 25 patients with unclear lesion areas or pathological results, the remaining 565 patients were matched at a ratio of 1:3 by using propensity score matching. A total of 25 EESCC patients with comorbid liver cirrhosis and 75 matched EESCC patients were ultimately included in the analysis. Parametric and nonparametric statistical methods were used to compare the differences between the two groups. The Kaplan-Meier method was used to create survival curves, and differences in survival curves were compared by the log-rank test.Among 25 patients with liver cirrhosis and 75 matched noncirrhotic patients, there were no significant differences in intraoperative bleedingESTD is technically feasible, safe, and effective for patients with EESCC and liver cirrhosis. EESCC patients with

Olanzapine is an antipsychotic drug used to treat patients with schizophrenia due to its lower incidence of extrapyramidal symptoms. Previous studies have shown that olanzapine activates AMP-activated protein kinase (AMPK), and induce autophagy in SH-SY5Y cell line. In this study, we investigated whether olanzapine protected against rotenone-induced neurotoxicity in PC12 cells. We showed that treatment with olanzapine increased the phosphorylation of AMPK in both dose- and time-dependent manners in PC12 cells. In addition, olanzapine activated autophagy and increased autophagic vacuoles. Furthermore, olanzapine pretreatment could protect PC12 cells from rotenone-induced apoptosis. Besides, olanzapine pretreatment could suppress the rotenone-induced depolarization of mitochondrial potential and thus protect the cells. Moreover, pretreatment with specific AMPK inhibitor compound C or with autophagy inhibitor 3-methyladenine impaired the protective effect of olanzapine on rotenone-treated PC12 cells. In summary, our results show for the first time that olanzapine ameliorates rotenone-induced injury by activating autophagy through AMPK pathway.

Excitotoxicity refers to the ability of excessive extracellular excitatory amino acids to damage neurons via receptor activation. It is a crucial pathogenetic process in neurodegenerative diseases. TP53 is confirmed to be involved in excitotoxicity. It is demonstrated that TP53 induced glycolysis and apoptotic regulator (TIGAR)-regulated metabolic pathway can protect against neuronal injury. However, the role of TIGAR in excitotoxicity and specific mechanisms is still unknown. In this study, an in vivo excitotoxicity model was constructed via stereotypical kainic acid (KA) injection into the striatum of mice. KA reduced TIGAR expression levels, neuroinflammatory responses and mitochondrial dysfunction. TIGAR overexpression could reverse KA-induced neuronal injury by reducing neuroinflammation and improving mitochondrial function, thereby exerting neuroprotective effects. Therefore, this study could provide a potential therapeutic target for neurodegenerative diseases.

Genetic studies have implicated both CD40 and tumour necrosis factor receptor-associated factor-1 (TRAF1) with rheumatoid arthritis (RA). CD40 signalling is known to be associated with TRAF1 expression, directly or indirectly; however, the detailed mechanisms of these interactions are not clear in RA, and in particular in fibroblast-like synoviocytes. This study aims to investigate this pathway and the role of nuclear factor-κB (NF-κB) in a mouse model of RA.We utilized the collagen-induced arthritis (CIA) model in DBA/1 mice. CD40, TRAF1, and NF-κB p65 were quantitated by enzyme-linked immunosorbent assay and immunohistochemistry in serum and tissue, respectively. Real-time polymerase chain reaction was applied to measure NF-κB-related gene expression, including cytokines [tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6)] and adhesion molecules [intercellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1)].The severity of arthritis by clinical and histological assessments peaked on day 35 and decreased thereafter. Serum levels of CD40, TRAF1, and NF-κB p65 paralleled this time-course. The tissue expression of the CD40, TRAF1, total NF-κB p65, and phospho-NF-κB p65 proteins, as well as NF-κB-related gene expression, including cytokines (TNFα, IL-6) and adhesion molecules (ICAM-1, VCAM-1), were markedly upregulated within 25-50 days after CIA.Our data identify a cellular/molecular mechanism of the CD40/TRAF1 signalling pathway involved in CIA: increased expression of CD40 and its adaptor TRAF1 proteins and activation of the NF-κB-dependent proinflammatory pathway. These correlations implicate possible mechanistic pathways in this model that may also operate in human RA and thus provide rationales for new therapeutic modalities.

Our previous studies confirm that exogenous reduced nicotinamide adenine dinucleotide phosphate (NADPH) exerts a neuroprotective effect in animal models of ischemic stroke, and its primary mechanism is related to anti-oxidative stress and improved energy metabolism. However, it is unknown whether nicotinamide adenine dinucleotide (NADH) also plays a neuroprotective role and whether NADPH is superior to NADH against ischemic stroke? In this study we compared the efficacy of NADH, NADPH, and edaravone in ameliorating brain injury and metabolic stress in ischemic stroke. Transient middle cerebral artery occlusion/reperfusion (t-MCAO/R) mouse model and in vitro oxygen glucose deprivation/reoxygenation (OGD/R) model were established. The mice were intravenously administered the optimal dose of NADPH (7.5 mg/kg), NADH (22.5 mg/kg), or edaravone (3 mg/kg) immediately after reperfusion. We showed that the overall efficacy of NADPH in ameliorating ischemic injury was superior to NADH and edaravone. NADPH had a longer therapeutic time window (within 5 h) after reperfusion than NADH and edaravone (within 2 h) for ischemic stroke. In addition, NADPH and edaravone were better in alleviating the brain atrophy, while NADH and NADPH were better in increasing the long-term survival rate. NADPH showed stronger antioxidant effects than NADH and edaravone; but NADH was the best in terms of maintaining energy metabolism. Taken together, this study demonstrates that NADPH exerts better neuroprotective effects against ischemic stroke than NADH and edaravone.

Ancylostomiasis is a fairly common small bowel parasite disease identified by capsule endoscopy (CE) for which a computer-aided clinical detection method has not been established. We sought to develop an artificial intelligence system with a convolutional neural network (CNN) to automatically detect hookworms in CE images. We trained a deep CNN system based on a YOLO-V4 (You Look Only Once-Version4) detector using 11236 CE images of hookworms. We assessed its performance by calculating the area under the receiver operating characteristic curve and its sensitivity, specificity, and accuracy using an independent test set of 10,529 small-bowel images including 531 images of hookworms. The trained CNN system required 403 seconds to evaluate 10,529 test images. The area under the curve for the detection of hookworms was 0.972 (95% confidence interval (CI), 0.967-0.978). The sensitivity, specificity, and accuracy of the CNN system were 92.2%, 91.1%, and 91.2%, respectively, at a probability score cut-off of 0.485. We developed and validated a CNN-based system for detecting hookworms in CE images. By combining this high-accuracy, high-speed, and oversight-preventing system with other CNN systems, we hope it will become an important supplement for detecting intestinal abnormalities in CE images. This trial is registered with ChiCTR2000034546 (a clinical research of artificial-intelligence-aided diagnosis for hookworms in small intestine by capsule endoscope images).

AimPrevious research recognizes that NADPH can produce reduced glutathione (GSH) as a coenzyme and produce ROS as a substrate of NADPH oxidase (NOX). Besides, excessive activation of glutamate receptors results in mitochondrial impairment. The study aims at spelling out the effects of NADPH and Mito-apocynin, a NOX inhibitor which specifically targets the mitochondria, on the excitotoxicity induced by Kainic acid (KA) and its mechanism.MethodsThe in vivo neuronal excitotoxicity model was constructed by stereotypically injecting KA into the unilateral striatum of mice. Administrated NADPH (i.v, intravenous) 30 min prior and Mito-apocynin (i.g, intragastric) 1 day prior, respectively, then kept administrating daily until mice were sacrificed 14 days later. Nissl staining measured the lesion of striatum and survival status of neurons. Cylinder test of forelimb asymmetry and the adhesive removal test reflected the behavioral deficit caused by neural dysfunction. Determined Total superoxide dismutase (T-SOD), malondialdehyde (MDA), and GSH indicated oxidative stress. Western blot presented the expression levels of LC3-II/LC3-I, SQSTM1/p62, TIGAR, and NOX4. Assessed oxygen consumption rate using High-Resolution Respirometry. In vitro, the MitoSOX Indicator reflected superoxide released by neuron mitochondria. JC-1 and ATP assay Kit were used to detect mitochondrial membrane potential (MMP) and energy metabolism, respectively.ResultsIn this study, we have successfully established excitotoxic model by KA in vivo and in vitro. KA induced decreased SOD activity and increased MDA concentration. KA cause the change of LC3-II/LC3-I, SQSTM1/p62, and TIGAR expression, indicating the autophagy activation. NADPH plays a protective role in vivo and in vitro. It reversed the KA-mediated changes in LC3, SQSTM1/p62, TIGAR, and NOX4 protein expression. Mito-apocynin inhibited KA-induced increases in mitochondrial NOX4 expression and activity. Compared with NADPH, the combination showed more significant neuroprotective effects, presenting more neurons survive and better motor function recovery. The combination also better inhibited the over-activated autophagy. In vitro, combination of NADPH and Mito-apocynin performed better in restoring mitochondria membrane potential.ConclusionIn summary, combined administration of NADPH and NOX inhibitors offers better neuroprotection by reducing NADPH as a NOX substrate to generate ROS. The combined use of NADPH and Mito-apocynin can better restore neurons and mitochondrial function through autophagy pathway.

Lung cancer is the leading cause of cancer-associated mortality worldwide. DNA damage-regulated autophagy modulator 1 (DRAM1) plays an important roles in autophagy and tumor progression. However, the mechanisms by which DRAM1 inhibits tumor growth are not fully understood. Here, we report that DRAM1 was decreased in nonsmall-cell lung carcinoma (NSCLC) and was associated with poor prognosis. We confirmed that DRAM1 inhibited the growth, migration, and invasion of NSCLC cells in vitro. Furthermore, overexpression of DRAM1 suppressed xenografted NSCLC tumors in vivo. DRAM1 increased EGFR endocytosis and lysosomal degradation, downregulating EGFR signaling pathway. On one side, DRAM1 interacted with EPS15 to promote EGFR endocytosis, as evidence by the results of proximity labeling followed by proteomics; on the other, DRAM1 recruited V-ATP6V1 subunit to lysosomes, thereby increasing the assemble of the V-ATPase complex, resulting in decreased lysosomal pH and increased activation of lysosomal proteases. These two actions of DRAM1 results in acceleration of EGFR degradation. In summary, these in vitro and in vivo studies uncover a novel mechanism through which DRAM1 suppresses oncogenic properties of NSCLC by regulating EGFR trafficking and degradation and highlights the potential value of DRAM1 as a prognostic biomarker in lung cancers.

Esophageal cancer is a common malignancy of digestive tract, and its prognosis is closely related to early diagnosis and treatment. In recent years, with the development of endoscopic diagnosis and treatment technology, especially the breakthrough progress of new endoscopic equipments, the detection rate of early esophageal cancer and precancerous lesions is significantly improved, and more and more patients with early esophageal cancer can be treated with minimally invasive endoscopic treatment. At present, endoscopic dissection has become the preferred treatment for early esophageal cancer and precancerous lesions. The advantages is not only less traumatic than traditional surgery, but also retain the physiological structure of esophagus, achieve accurate postoperative pathology and better postoperative quality of life of patients. However, endoscopic minimally invasive treatment in progress also face new problems to be solved, such as how to deal with multifocal esophageal lesions, how to estimate esophageal lesions infiltration depth and lymph node metastasis risk accurately, how to understand the discrepancy between biopsy and postoperative pathology, how to deal with the positive resection margins as well as intraoperative and postoperative complications, how to manage endoscopic treatment of the special subpopulation of the patients, whether and when additional radical surgery should be provided to the patients with non-curative endoscopic treatment, and so on. Aiming to the above problems and the purpose to improve the prognosis and the quality of life of esophageal cancer patients, this topic includes a series studies to explore standardized treatment scheme and management strategy for postoperative complications in the endoscopic treatment of early esophageal cancer and precancerious lesions.

To explore endoscopic characteristics and pathological changes of esophageal low-grade intraepithelial neoplasm (LGIN) as well as its risk factors.A total of 201 LGIN lesions from 169 cases were included from January 2009 to August 2017. The endoscopic characteristics and pathological changes were analysis. Logistic regression analysis was used to analyze the risk factors of LGIN. The endoscopic morphologic findings of esophageal mucosa lesions and the pathological findings of simple inflammatory lesions were enrolled as controls.LGIN occurred more common in elderly patients, the ratio of male to female was 2.5∶1. The maximum transverse and the maximum longitudinal diameter (MLD) were (0.9±0.8) cm,(1.4±1.3) cm, respectively. The most common location of lesion was in the middle segment of esophagus (52.2%). The morphological types of lesions were dominantly 0-Ⅱb (45.8%) and 0-Ⅱa (31.8%). There were 42 LGIN lesions with reflux esophagitis. Multiple dysplastic lesions accounted for 57.4%. After (10.3±12.1) months follow-up, 58.2% lesions were pathological reversal with 24.9% (50/201) of the lesion completely disappeared, and 28.9% lesions had no pathological changes, but 12.9% (26/201) lesions progressed to high-grade intraepithelial neoplasia and invasive cancer. Multivariate analysis indicated that age (compared to45 years old) and longitudinal diameter of the lesion (compared to ≤0.5 cm) were independent risk factors for LGIN. The risk of esophageal LGIN in lesions with MLD0.5-1 cm was 1.96 times higher than that in lesions with MLD ≤ 0.5 cm.The MLD of esophageal mucosal lesions0.5 cm and age45 years old may increase the possibility of esophageal LGIN. Close follow-up is required for LGIN lesions with MLD1 cm.

To evaluate the efficacy of oral corticosteroids in preventing esophageal stenosis after large area esophageal endoscopic submucosal tunnel dissection (ESTD).The patients undertook esophageal ESTD were included from January 2014 to January 2018. The inclusion criteria was single lesion of esophageal early esophagus cancer with the extent more than 3/4 of circumferential degree. According to the inclusion time, the patients were divided into the trial group (ESTD + oral corticosteroids) and the control group (simple ESTD). The incidence of the total esophageal stenosis, intractable esophageal stenosis, the remission rate of dysphagia and the period from the dysphagia present were observed and compared in the two groups.A total of 101 cases of esophageal ESTD patients were included. There were 48 cases in the trial group, 28 cases of male and 20 cases of female, with an average age of (62.98±7.52) years; 53 cases in the control group, 28 cases of male and 25 cases of female, with an average age of (62.67±8.04) years. The rate of intractable esophageal stenosis in the trial group was lower than that in the control group (6.25% vs. 20.75%,Oral corticosteroids can effectively reduce the degree of esophageal stenosis after large area ESTD, as well as the incidence of intractable esophageal stenosis and the number of endoscopic treatment in non-refractory esophageal stenosis patients.

To assess the accuracy of endoscopic ultrasound (EUS) and magnifying endoscopy with narrow-band imaging (ME-NBI) in evaluating the invasion depth of early esophageal carcinoma.Patients who underwent endoscopic resection for early esophageal cancer from March 2013 to October 2017 were enrolled. The EUS and ME-NBI results were compared with the pathology results.A total of 392 lesions from 333 patients were assessed, including 83 mild and moderate dysplasia, 72 severe dysplasia, 235 squamous cell carcinoma, and 2 adenosquamous carcinoma. About 308 lesions were given EUS only, 7 had ME-NBI only, 77 underwent both EUS and ME-NBI. EUS resulted in a 43.9% accuracy for the 385 lesions, with poor consistency (ME-NBI has higher accuracy than EUS in evaluating the invasion depth of early esophageal carcinoma.

Parkinson’s disease (PD) is the second most common neurodegenerative disorder characterized by the selective loss of dopaminergic neurons in substantia nigra pars compacta (SNpc). Although the pathogenic mechanism underlying PD remains largely unknown, decreased nigral glutathione (GSH) in postmortem brains of PD patients supports the presence of oxidative stress in PD. We found that Nicotinamide adenine dinucleotide phosphate (NADPH), which is important for maintaining the level of GSH, protected dopaminergic (DA) neurons from neurotoxicity of MPTP/MPP(+). In the present study, NADPH prevented DA neurons from MPTP toxicity with increased GSH and decreased reactive oxygen species (ROS) levels in the ventral midbrain of mice, and improved motor activity. Our present results demonstrated that NADPH inhibited the phosphorylation of p38MAPK, decreased the level of TP53 protein, and inhibited TP53 nuclear translocation in DA neurons of SNpc and in MES23.5 cells. Furthermore, NADPH decreased the protein level of TP53 target gene, Bax, cleavage of PARP, and nuclei condensation. Taken together, NADPH abrogated MPTP-induced p38MAPK phosphorylation, TP53 nuclear translocation, and Bax induction, and finally, MPTP/MPP(+)-induced apoptosis of DA neurons. This study suggests that NADPH may be a novel therapeutic candidate for PD.

Both reduced nicotinamide adenine dinucleotide phosphate (NADPH) and β-nicotinamide adenine dinucleotide hydrate (NAD+) have been reported to have potent neuroprotective effects against ischemic neuronal injury. Both NADPH and NAD+ are essential cofactors for anti-oxidation and cellular energy metabolism. We investigated if combined NADPH and NAD+ could offer better neuroprotective effects on cellular and animal models of ischemic stroke. In vitro studies with primary cultured neurons demonstrated that NAD+ was effective in protecting neurons against oxygen–glucose deprivation/reoxygenation (OGD/R) injury when given during the early time period of reoxygenation. In vivo studies in mice also suggested that NAD+ was effective for ameliorating ischemic brain damage when administered within 2 h after reperfusion. The combination of NADPH and NAD+ provided not only greater beneficial effects but also larger therapeutic window in both cellular and animal models of stroke. The combination of NADPH and NAD+ significantly increased the levels of adenosine triphosphate (ATP) and reduced the levels of reactive oxygen species (ROS) and oxidative damage of macromolecules. Furthermore, the combined medication significantly reduced long-term mortality, improved the functional recovery, and inhibited signaling pathways involved in apoptosis and necroptosis after ischemic stroke. The present study indicates that the combination of NAD+ and NADPH can produce greater therapeutic effects with smaller dose of NADPH; on the other hand, NADPH can significantly prolong the therapeutic window of NAD+. The current results suggest that the combination of NADPH and NAD+ may provide a novel effective therapy for ischemic stroke.

Purpose To investigate the effects and mechanisms of NADPH on Kainic acid (KA)-induced excitotoxicity. Methods KA, a non-N-methyl- d -aspartate glutamate receptor agonist, was exposed to adult SD rats via intrastriatal injection and rat primary cortical neurons to establish excitotoxic models in vivo and in vitro, respectively. To determine the effects of NADPH on KA-induced excitotoxicity, neuronal survival, neurologically behavioral score and oxidative stress were evaluated. To explore the mechanisms of neuroprotective effects of NADPH, the autophagy-lysosome pathway related proteins were detected. Results In vivo, NADPH (1 mg/kg or 2 mg/kg) diminished KA (2.5 nmol)-induced enlargement of lesion size in striatum, improved KA-induced dyskinesia and reversed KA-induced activation of glial cells. Nevertheless, the neuroprotective effect of NADPH was not significant under the condition of autophagy activation. NADPH (2 mg/kg) inhibited KA (2.5 nmol)-induced down-regulation of TP-53 induced glycolysis and apoptosis regulator (TIGAR) and p62, and up-regulation of the protein levels of LC3-II/LC3-I, Beclin-1 and Atg5. In vitro, the excitotoxic neuronal injury was induced after KA (50 μM, 100 μM or 200 μM) treatment as demonstrated by decreased cell viability. Moreover, KA (100 μM) increased the intracellular levels of calcium and reactive oxygen species (ROS) and declined the levels of the reduced form of glutathione (GSH). Pretreatment of NADPH (10 μM) effectively reversed these changes. Meanwhile NADPH (10 μM) inhibited KA (100 μM)-induced down-regulation of TIGAR and p62, and up-regulation of the ratio of LC3-II/LC3-I, Beclin-1, Atg5, active-cathepsin B and active-cathepsin D. Conclusions Our data provide a possible mechanism that NADPH ameliorates KA-induced excitotoxicity by blocking the autophagy-lysosome pathway and up-regulating TIGAR along with its antioxidant properties.

Background Macroautophagy (hereafter autophagy) is a tightly regulated process that delivers cellular components to lysosomes for degradation. Damage-regulated autophagy modulator 1 (DRAM1) induces autophagy and is necessary for p53-mediated apoptosis. However, the signalling pathways regulated by DRAM1 are not fully understood. Methods HEK293T cells were transfected with FLAG-DRAM1 plasmid. Autophagic proteins (LC3 and p62), phosphorylated p53 and the phosphorylated proteins of the class I PI3K-Akt-mTOR-ribosomal protein S6 (rpS6) signalling pathway were detected with Western blot analysis. Cellular distribution of DRAM1 was determined with immunostaining. DRAM1 was knocked down in HEK293T cells using siRNA oligos which is confirmed by quantitative RT-PCR. Cells were serum starved for 18 h after overexpression or knockdown of DRAM1 to decrease the rpS6 activity to the basal level, and then the cells were stimulated with insulin growth factor, epidermal growth factor or serum. rpS6 phosphorylation and rpS6 were detected with Western blotting. Similarly, after overexpression or knockdown of DRAM1, phosphorylation of IGF-1Rβ and IGF-1R were examined with Western blotting. Cell viability was determined with CCK-8 assay and colony formation assay. Finally, human cancer cells Hela, SW480, and HCT116 were transfected with the FLAG-DRAM1 plasmid and phosphorylated rpS6 and rpS6 were detected with Western blot analysis. Results DRAM1 induced autophagy and inhibited rpS6 phosphorylation in an mTORC1-dependent manner in HEK293T cells. DRAM1 didn’t affect the phosphorylated and total levels of p53. Furthermore, DRAM1 inhibited the activation of the PI3K-Akt pathway stimulated with growth factors or serum. DRAM1 was localized at the plasma membrane and regulate the phosphorylation of IGF-1 receptor. DRAM1 decreased cell viability and colony numbers upon serum starvation. Additionally, DRAM1 inhibited rpS6 phosphorylation in several human cancer cells. Conclusions Here we provided evidence that DRAM1 inhibited rpS6 phosphorylation in multiple cell types. DRAM1 inhibited the phosphorylation of Akt and the activation of Akt-rpS6 pathway stimulated with growth factors and serum. Furthermore, DRAM1 regulated the activation of IGF-1 receptor. Thus, our results identify that the class I PI3K-Akt-rpS6 pathway is regulated by DRAM1 and may provide new insight into the potential role of DRAM1 in human cancers.

Autophagy has been reported to play protective and pathogenetic roles in cerebral ischemia/reperfusion (I/R)-induced neuronal injury. Our previous studies have shown that TP53-induced glycolysis and apoptosis regulator (TIGAR) ameliorates I/R-induced brain injury and reduces anti-cancer drug-induced autophagy activation. However, if TIGAR plays a regulatory role on autophagy in cerebral I/R injury is still unclear. The purpose of the present study is to investigate the role of TIGAR on I/R-induced autophagy activation and ischemic neuronal injury in vivo and in vitro stroke models using TIGAR-transgenic (tg-TIGAR) mice and TIGAR-knockout (ko-TIGAR) mice. The present study confirmed that autophagy was activated after I/R. Overexpression of TIGAR in tg-TIGAR mice significantly reduced I/R-induced autophagy activation and alleviated brain damage, while knockout of TIGAR in ko-TIGAR mice enhanced I/R-induced autophagy activation and exacerbated brain injury in vivo and in vitro. The different activity of autophagy in tg-TIGAR and ko-TIGAR primary neurons after OGD/R were largely reversed by knockdown or re-expression of TIGAR in these neurons. The autophagy inhibitor 3-methyladenine (3-MA) partly prevented exacerbation of brain damage induced by ko-TIGAR, whereas the autophagy inducer rapamycin partially abolished the neuroprotective effect of tg-TIGAR. Knockout of TIGAR reduced the levels of phosphorylated mTOR and S6KP70, which were blocked by 3-MA and NADPH after I/R and OGD/R in vivo and in vitro, respectively. Overexpression of TIGAR increased the levels of phosphorylated mTOR and S6KP70 under OGD/R condition, this enhancement effect was suppressed by rapamycin. In conclusion, our current data suggest that TIGAR protected against neuronal injury partly through inhibiting autophagy by regulating the mTOR-S6KP70 signaling pathway.

This study reports the outcomes of endoscopic submucosal single-tunnel dissection or endoscopic submucosal multi-tunnel dissection for the treatment of esophageal neoplastic lesions of at least three-quarters of the esophageal circumference, including circumferential superficial esophageal neoplastic lesions. From July 2014 to February 2018, a total of 124 lesions underwent endoscopic submucosal tunnel dissection at our hospital. One to four submucosal tunnels were created in the oral to anal direction. Of the 124 lesions, there were 83 noncomplete circumferential lesions and 41 circumferential lesions. Endoscopic submucosal single-tunnel dissection was performed in 54 patients, two-tunnel dissection in 43 patients, three-tunnel dissection in 19 patients, and four-tunnel dissection in 8 patients. The mean dissection speed was 22.8±12.7 mm2/min. En bloc dissection was achieved in all lesions, and the R0 resection rate was 70.2 percent. No matter how large the lesion area was, there were no significant differences in the dissection speed and the R0 resection rate when lesions were at least three-quarters of the esophageal circumference. Esophageal stricture was observed in 54 patients and was relieved by placement of a retrievable metal stent or by endoscopic water balloon dilation. No recurrence was noted after 19.1±12.4 months of follow-up. Our large sample size study showed that endoscopic submucosal tunnel dissection showed effectiveness and safety for the treatment of large superficial esophageal neoplastic lesions at least three-quarters of the esophageal circumference, including circumferential superficial esophageal neoplastic lesions.

Summary Aim To study the effects of exercise on lysosomal functions. Methods Mouse exercise model was established and wheel running was scheduled as 18 rpm (14:00‐17:00), 5 d/wk, for 8 weeks. Mice were injected EX527 to inhibit SIRT1 activity. The protein level was assayed with Western blot and immunofluorescence histochemistry. The transmission electron microscopic examination was used to show the structure of lysosome and mitochondria. Results Exercise promoted the nuclear translocation of TFEB in the cortex which upregulated the transcription of genes associated with autophagy and lysosome. Exercise directly activated autophagy/lysosome system via up‐regulating of AMPK‐SIRT1 signaling. The SIRT1 inhibitor EX527 decreased TFEB regulated gene transcription but had little effect on the nuclear translocation of TFEB. In addition, long‐term exercise showed more significant effects on activation of lysosomes biogenesis compared with the short‐term exercise and trehalose, a classical autophagy activator in the mTOR‐independent pathway. Conclusion Running exercise activates lysosomal function in the brain through AMPK‐SIRT1‐TFEB pathway.

Background and Aims Endoscopic submucosal tunnel dissection (ESTD) has been used for dissection of large esophageal neoplastic lesions, but there are still some technical problems in treating circumferential superficial esophageal neoplastic lesions. This study aimed to assess the efficacy and safety of endoscopic submucosal multi-tunnel dissection (ESMTD) for circumferential superficial esophageal neoplastic lesions in selected patients followed up for 1 to 12 months. Methods From July 2014 to February 2015, the first series of 7 consecutive patients with circumferential superficial esophageal neoplastic lesions underwent ESMTD at our endoscopic center. The macroscopic types were classified according to the Paris endoscopic classification of superficial neoplastic lesions. Results The average length of lesions was 6.1 cm in 7 selected patients. The operative time ranged from 69 to 169 minutes (mean 121 minutes). En bloc dissection with negative basal margins was achieved in all lesions without serious intraoperative adverse events. Esophageal stricture was observed in all patients 1 to 3 months after the operation and was relieved after a retrievable metal stent was placed or esophageal water balloon dilatation was performed; however, one patient died of cerebral infarction 2 months after ESMTD. Conclusions ESMTD is feasible, safe, and effective for the treatment of circumferential superficial esophageal neoplastic lesions in select patients.

Background and Purpose— Our previous study has defined a role of TP53-induced glycolysis and apoptosis regulator in neuroprotection against ischemic injury through increasing the flow of pentose phosphate pathway. We hypothesized that the pentose phosphate pathway product nicotinamide adenine dinucleotide phosphate (NADPH) could be a novel drug for treatment of ischemic stroke. Methods— The NADPH was given before, at the onset, or after stroke onset with single or repeated intravenous (mice and rats) or intraperitoneal injections (monkey). The short- and long-term therapeutic effects of NADPH were evaluated in male adult ICR mice (total=614) with transient middle cerebral artery occlusion, in male adult Sprague–Dawley rats (total=114) with permanent middle cerebral artery occlusion, and in male adult rhesus monkey (total=12) with thrombotic middle cerebral artery occlusion. Results— Administration of NADPH led to a dramatic increase in the levels of ATP and reduced form of glutathione, whereas it decreased the levels of reactive oxygen species. NADPH significantly reduced infarct volume, improved poststroke survival, and recovery of neurological functions in mouse and rat models of stroke. Robust neuroprotection of a single dose of NADPH was seen when it was administered within 5 hours after reperfusion; however, repeat administration of NADPH twice a day for 7 days starting 24 hours after the onset of stroke also offered therapeutic effects. Pretreatment with NADPH also significantly improved the outcome of stroke insult. Conclusions— Administration of exogenous NADPH significantly protected neurons against ischemia/reperfusion-induced injury in 2 rodent stroke models. Thus, NADPH might be a promising drug candidate for treatment of ischemic stroke.

TP53-induced glycolysis and apoptosis regulator (TIGAR) is a p53 target protein that plays critical roles in glycolysis and redox balance. Accumulating evidence shows that TIGAR is highly expressed in cancer. TIGAR redirects glycolysis and promotes carcinoma growth by providing metabolic intermediates and reductive power derived from pentose phosphate pathway (PPP). The expression of TIGAR in cancer is positively associated with chemotherapy resistance, suggesting that TIGAR could be a novel therapeutic target. In this review, we briefly presented the function of TIGAR in metabolic homeostasis in normal and cancer cells. Finally, we discussed the future directions of TIGAR research in cancer.

DRAM1 (DNA damage-regulated autophagy modulator 1) is a transmembrane protein that predominantly localizes to the lysosome but is also found in other membranous organelles; however, its function in these organelles remains largely unknown. We found that DRAM1 was partially located in the Golgi apparatus, and knockdown of DRAM1 caused fragmentation of the Golgi apparatus in cells. The phenomenon of fragmented Golgi was not related to microtubule organization, and there was no direct interaction between DRAM1 and Golgi structural proteins (ARF1, GM130, syntaxin 6 and GRASP55). Moreover, Golgi-targeting DRAM1 failed to rescue the fragmentation of Golgi in DRAM1-deficient cells. The transport of ts045-VSVG-GFP, an indicator of movement from the Golgi apparatus to the plasma membrane, was delayed in DRAM1-knockdown cells. Moreover, the trafficking of CI-MPR from the plasma membrane to the Golgi was also impeded in DRAM1-knockdown cells. These results indicated that DRAM1 regulated the structure of the Golgi apparatus and affected Golgi apparatus-associated vesicular transport.

Background: Endoscopic submucosal dissection (ESD) has been used to treat early stage esophageal cancer, but reports about additional esophagectomy after ESD and postoperative outcomes are lacking. Complete removal of cancer tissue together with lymph nodes was the advantage of esophagectomy; however, invasiveness, organ loss, postoperative complications, and worse postoperative quality of life were serious disadvantages. The purpose of this study was to find the clear indication of additional esophagectomy after ESD, and help the other patients avoid excessive surgery. Methods: We reviewed the clinicopathologic data and outcomes consecutive patients who had esophageal cancer confirmed by endoscopic biopsy and who were treated with ESD and subsequent esophagectomy between October 2011 and December 2016 in our department. The esophagectomy necessity following ESD was defined and the groups with necessity (+) vs . (−) were compared retrospectively. The esophagectomy necessity outcomes were retrospectively analyzed to judge whether the surgery option was correct. Results: Total 214 patients with esophageal and esophagogastric cancer have undergone ESD treatment in our center, of which 32 patients (23 men and 9 women; mean age, 60±8 years) ultimately required esophagectomy after ESD. All patients had complete resection (R0) from esophagectomy. Postoperative TNM staging included TisN0M0 (6 patients), T1aN0M0 (6 patients), T1bN0M0 (18 patients), T1bN1M0 (1 patient), and T2N3M0 (1 patient). Necessity of esophagectomy after ESD was associated with residual margin status. There was a significant difference in ESD specimen margin status between the esophagectomy necessity (+) vs . (−) groups (positive/negative margin: 8/3 vs . 2/9 patients; P=0.03). Esophagectomy should be delayed at least 30 days after ESD to enable resolution of esophageal edema (P=0.017) (206±68 vs . 163±56 mL, P=0.057). Median follow-up was 16.8 months (range, 11.2–54.5 months); 3 patients were lost to follow-up (9%) and 1 patient died of metastasis after esophagectomy. All other patients were alive with excellent postoperative disease-free survival. Conclusions: Indications for esophagectomy after ESD include ESD failure, cancer recurrence, esophageal rupture, esophageal stricture refractory to endoscopic dilation, and residual tumor at the ESD specimen margin. Stage T1b alone is not an indication for esophagectomy. According to our study, we recommend that esophagectomy should be delayed ≥30 dafter ESD unless urgent esophagectomy is indicated.

It is generally believed that oxidative stress and neuroinflammation are implicated in the pathogenesis of Parkinson's disease (PD). Reduced nicotinamide adenine dinucleotide phosphate (NADPH) has been demonstrated to have potent neuroprotective effects against oxidative stress. In the present research, we investigated if NADPH could offer neuroprotection by inhibiting glia-mediated neuroinflammation induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a mechanism contributing to PD pathogenesis. The current data demonstrated that MPTP/MPP+ increased levels of reactive oxygen species (ROS), activated glial cells, and inflammasome proteins in the substantia nigra (SNpc), in addition to inducing the nuclear translocation of nuclear factor-κB (NF-κB) and phosphorylation of p38 MAPK. These responses were inhibited by supplementation of exogenous NADPH. Moreover, NADPH effectively decreased MPP+-induced excessive production of ROS, p38 phosphorylation and inflammatory protein of Cyclooxygenase2 (COX2) in cultured microglial BV-2 cells in vitro studies. Similarly, the p38 MAPK inhibitor SB203580 suppressed the upregulation of MPP+-induced p38 phosphorylation and COX2 protein levels. Co-culture of neuronal cells with MPP+-primed BV-2 cells increased the levels of tumor necrosis factor-alpha (TNF-α) and induced cell death of neuronal cells. These effects were diminished by TNF-α neutralizing antibody and NADPH. NADPH reduced motor dysfunction and the loss of dopaminergic (DA) cells induced by MPTP. Therefore, the present study demonstrates that NADPH protects DA neurons by inhibiting oxidative stress and glia-mediated neuroinflammation both in vitro and in vivo, thus suggesting a potential of clinical application for PD and other neurodegenerative diseases.

Previous studies found that reduced nicotinamide adenine dinucleotide phosphate (NADPH) protected neurons against ischemia/reperfusion-induced injury. In addition to ROS reduction and ATP increment, preliminary data suggested that NADPH inhibited ADP and thrombin-induced platelet aggregation. As the effect of NADPH on platelet function was not reported by other investigators, the actions of NADPH on platelet function and mechanisms of actions were investigated in the present study. In vitro studies, the effects of different concentrations of NADPH on platelet aggregation induced by ADP (10 μM), thrombin (0.05 U/mL) or AA (50 μM) were determined. The results showed that NADPH could inhibit platelet aggregation induced by ADP, thrombin or AA in a concentration dependent manner. When the inhibitory effects of NAD+, NADH, NADP+ and NADPH on platelet aggregation were compared, NADPH demonstrated the relatively best effect on platelet aggregation. In vivo studies, the effects of NADPH on platelet aggregation, tail bleeding time, coagulation response and ferric chloride-induced thrombosis were determined in mice or rats. The maximum aggregation rate of platelets of rats injected with NADPH (5 mg/kg) was lower than platelets from control rats. NADPH transiently prolonged tail bleeding time in mice at 30 min after the injection of NADPH (7.5 mg/kg), while aspirin (15 mg/kg) significantly prolonged the tail bleeding time in mice at all time points examined. NADPH (5 mg/kg), as well as aspirin (10 mg/kg), had no effect on coagulation response in rats. Using a FeCl3-induced abdominal aorta injury thrombosis model, administration of NADPH (5 mg/kg) significantly delayed the onset of vessel occlusion, while aspirin (10 mg/kg) almost completely prevented the vessel occlusion. With microscopic examination the thrombi in injured vessel sections of rats received NADPH were much smaller and less dense than that of rats received vehicle treatment. ADP induced an increase in phosphorylation of p38 and the effect was markedly inhibited by the p38 inhibitor SB203580. Similarly, NADPH also inhibited ADP-induced phosphorylation of p38. Similar to NADPH, SB203580 robustly inhibited ADP- and thrombin-induced platelet aggregation. In addition, NADPH also reduced ADP-induced increases in ROS in platelets. The current results demonstrated that NADPH inhibited platelet aggregation, oxidative stress and p38 phosphorylation, suggesting that NADPH might be a novel compound for management of high risk of cardiovascular disease.

Both reduced nicotinamide adenine dinucleotide phosphate (NADPH) and β-nicotinamide adenine dinucleotide hydrate (NAD

To investigate the role of PIK3CA oncogene in tumorigenesis and development of esophageal cancer in Chinese patients at the levels of genetic mutation and epigenetics.Seventy six esophageal tumor samples and corresponding adjacent normal tissues were collected, and the genomic DNA was extracted. Mutations in the 9th and 20th exons of PIK3CA gene were detected using conventional sequencing. PIK3CA methylation rates in two selected CpG islands (CpG island 1 and 2) were detected using sub-bisulfate modified sequencing. P110α and pAKT expression levels were detected with Western blotting.In PIK3CA gene of the tumor tissues, G1633C (E545Q) mutation was detected in the 9th exon with a rate of 3.95% (3/76), whereas mutation was not found in the 20th exon. Nor mutation did occur in PIK3CA gene of the adjacent normal tissues. The methylation rate of the CpG island 1 had no significant difference between the tumor and adjacent tissues (0.77%±0.009% vs 0.89%±0.008%), but the methylation rate of the CpG island 2 in the esophageal tumors was significantly lower than that in the adjacent tissues (6.00%±2.80% vs 10.45%±5.51%). Furthermore, the rate of methylation of the CpG island 2 in TNM stage III and IV esophageal cancer (3.84%±2.08%) was significantly lower than in stage I (8.52%±2.55%) and stage II (6.42%±2.36%). PIK3CA gene hypomethylation in esophageal cancer was significantly correlated with high expression of p110α.PIK3CA gene hypomethylation plays a key role in the tumorigenesis and development of esophageal cancer in Chinese patients, while the mutations of PIK3CA gene have little effect on the development of esophageal cancer.

To analysis the risk factors of lymph node metastasis (LNM) in superficial esophageal squamous cell carcinoma (SESCC).The clinical data and pathological results of 344 SESCC patients, who underwent surgical treatments between January 2009 and December 2013 in West China Hospital, Sichuan University, were analyzed retrospectively. Clinicopathologic characteristics were compared between different histological types, and their possible relationships with LNM were explored by univariate and multivariate analysis.There were no LNM found in the patients with tumor limited to the mucous, tumor diameter3 cm, highly and moderate differentiated SESCC. Univariate analysis showed that tumor diameter (P=0.004), depth of tumor invasion (P=0.009), histological type (P=0.030) and lymphatic involvement (P=0.002) were correlated with LNM. Multivariate analysis revealed that tumor diameter (P=0.007), depth of tumor invasion (P=0.003), histological type (P=0.010) and lymphatic involvement (P0.001) were independent risk factors of LNM.To the patients with low risk of LNM, such as tumor limited to the mucous, tumor diameter3 cm, and highly and moderate differentiation, endoscopic excision may be considered as an absolute indications.

To study the efficiency and complications of endoscopic submucosal dissection (ESD) and endoscopic submucosal tunnel dissection (ESTD) in the treatment of large esophageal mucosal lesions.The clinical data were collected from the patients who received ESD or ESTD for the treatment of early esophageal lesions in our hospital during January 1, 2014 to July 15, 2015, including the en bloc resection rate, curative resection rate, postoperative complication rate, the risk factors of complications were explored by univariate and multivariate analysis.A total of 50 patients were involved in the study, ESD or ESTD were performed successfully in 53 times, including 6 cases of ESD,47 cases of ESTD, The average age was (61.9 +/- 6.8) yr., the average operating time was (83.57 +/- 32.33) min, the average dissected lesion was (14.82 +/- 3.18) cm2, En bloc resection rate was 94.34%, the curative rate was 84.90%. There were 1 case of bleeding, 1 case of perforation, 45 (84.90%) cases of fever, 13 cases (24.53%) of esophageal stricture. The severity of stricture was associated with the operation time Codds ratio (OR) = 1.040, 95% confidence interal (CI): 1.007-1.075) and esophageal circumference (OR=9.972, 95% CI: 1.221-81.416). The residual resection margin appeared in 8 patients, and the lesion area (OR=1.145, 95% CI: 1.013-1.294) was the only risk factor.ESD and ESTD are safe and effective in the treatment of early esophageal lesions, but seems have relatively high incidence of esopgageal stricture and residual resection margin in the treatment of large esophageal lesions.

To determine the influence of location, depth and size of upper gastrointestinal (GI) submucosal tumors (SMTs) on the success of submucosal tunneling endoscopic resection (STER).Patient records of 31 cases with upper GI SMTs who had STER between Jan. 1, 2014 and June 30, 2015 in West China Hospital of Sichuan University were retrieved. The success of STER was determined by its efficiency, complete resection rate, and incidence of complications.Of the 31 cases, 29 were treated successfully, with an average of (13.76 +/- 9.70) min and (32.00 +/- 27.35) min for tunnel formation of esophageal and stomach mucosal tumors respectively (P = 0.045). The 2 unsuccessful cases were gastric tumors. SMTs resection for mucous layer and muscularis propria took (17.50 +/- 9.06) min and (36.24 +/- 15.68) min, respectively (P=0.004). SMTs resection for tumors diameter2.0 cm andor = 2.0 cm took (25.78 +/- 12.13) min and (39.73 +/- 19.23) min, respectively (P=0.023). Six cases of gastric tumors from muscularis propria had complications (19.4%) during or after surgery.Location, depth and size of upper GI SMTs has implications on duration of different STER stages, which may determine complete resection rate and incidence of complications.

To assess the cytotoxic effect of crotoxin (CrTX), a potent neurotoxin extracted from the venom of the pit viper Crotalus durissus terrificus, in human lung adenocarcinoma A549 cells and investigated the underlying mechanisms.A549 cells were treated with gradient concentrations of CrTX, and the cell cycle and apoptosis were analyzed using a flow cytometric assay. The changes of cellular effectors p53, caspase-3 and cleaved caspase-3, total P38MAPK and pP38MAPK were investigated using Western blot assays. A549 xenograft model was used to examine the inhibition of CrTX on tumor growth in vivo.Treatment of A549 cells with CrTX (25-200 μg/mL) for 48 h significantly inhibited the cell growth in a dose-dependent manner (IC(50)=78 μg/mL). Treatment with CrTX (25 μg/mL) for 24 h caused G1 arrest and induced cell apoptosis. CrTX (25 μg/mL) significantly increased the expression of wt p53, cleaved caspase-3 and phospho-P38MAPK. Pretreatment with the specific P38MAPK inhibitor SB203580 (5 μmol/L) significantly reduced CrTX-induced apoptosis and cleaved caspase-3 level, but G(1) arrest remained unchanged and highly expressed p53 sustained. Intraperitoneal injection of CrTX (10 μg/kg, twice a week for 4 weeks) significantly inhibited A549 tumor xenograft growth, and decreased MVD and VEGF levels.CrTX produced significant anti-tumor effects by inducing cell apoptosis probably due to activation of P38MAPK and caspase-3, and by cell cycle arrest mediated by increased wt p53 expression. In addition, CrTX displayed anti-angiogenic effects in vivo.

Background Adverse events such as perforation and bleeding during endoscopic submucosal dissection (ESD) of superficial esophageal neoplasms have been well acquainted. However, Mallory Weiss Tear (MWT) during esophageal ESD is still under obscurity. Therefore, this study was carried out to elucidate the incidence and risk factors for MWT during esophageal ESD. Methods Patients received ESD for superficial esophageal neoplasms from June, 2014 to July 2017 in our institution were retrospectively analyzed. Clinicopathological characteristics of the patients were collected. Patients were allocated into MWT group or non-MWT group based on the presence of MWT occurred during ESD. The incidence of MWT was determined and the risk factors for MWT were identified. Results A total of 337 patients with 373 lesions treated by ESD were analyzed. 20 patients developed MWT during ESD (5.4%). Multivariate analysis identified female gender (OR = 5.36, 95%CI: 1.47–19.50, P = 0.011) and procedure time (OR = 1.01, 95%CI: 1.00–1.02, P = 0.048) were independent risk factors for MWT during ESD. The characteristics of the esophageal lesions had no influence on the development of MWT. The vast majority of patients with MWT did not receive endoscopic treatment for the laceration and seven of the MWT patients received endoscopic hemostasis using APC, hemoclips, or hemoclips combined with endoloop. All the patients recovered satisfactorily without surgery for the laceration. We believe the placement of gastrointestinal decompression tube and the use of PPI and hemostatic drugs after ESD contributed to the hemostasis and mucosal healing. Conclusion The incidence of MWT during esophageal ESD was much higher than expected. Although most cases had a benign course, fetal conditions such as massive hemorrhage may also occur. We recommend an inspection of the stomach after ESD procedure in order to take timely management in case of bleeding MWT or even perforation outside the procedure region. Disclosure All authors have declared no conflicts of interest.

The present study sought to investigate mechanisms by which p53 induction contributes to excitotoxic neuronal injury. Rats were intrastriatally administered the N-methyl-D-aspartate (NMDA) receptor agonist quinolinic acid (QA), the changes in the expression of p53 and its target genes involved in apoptosis and autophagy, including p53-upregulated modulator of apoptosis (PUMA), Bax, Bcl-2, damage-regulated autophagy modulator (DRAM) and other autophagic proteins including microtubule-associated protein 1 light chain 3 (LC3) and beclin 1 were assessed. The contribution of p53-mediated autophagy activation to apoptotic death of striatal neurons was assessed with co-administration of the nuclear factor-kappaB (NF-kappaB) inhibitor SN50, the p53 inhibitor Pifithrin-alpha (PFT-alpha) or the autophagy inhibitor 3-methyladenine (3-MA). The increased formation of autophagosomes and secondary lysosomes were observed with transmission electron microscope after excitotoxin exposure. QA induced increases in the expression of p53, PUMA, Bax and a decrease in Bcl-2. These changes were significantly attenuated by pre-treatment with SN50, PFT-alpha or 3-MA. SN50, PFT-alpha or 3-MA also reversed QA-induced upregulation of DRAM, the ratio of LC3-II/LC3-I and beclin 1 protein levels in the striatum. QA-induced internucleosomal DNA fragmentation and loss of striatal neurons were robustly inhibited by SN50, PFT-alpha or 3-MA. These results suggest that overstimulation of NMDA receptors can induce NF-kappaB-dependent expression of p53. p53 participates in excitotoxic neuronal death probably through both apoptotic and autophagic mechanisms.

In vivo administration of the mitochondrial inhibitor 3-nitropropionic acid (3-NP) produces striatal pathology mimicking Huntington's disease (HD). However, the mechanisms of cell death induced by metabolic impairment are not fully understood. Previous studies showed that 3-NP triggered p53-depedent autophagy activation and cell death. The present study investigated the contribution of the Bcl-2 signaling pathway to autophagy activation and cell death induced by 3-NP. Rat striatum was intoxicated with 3-NP by stereotaxic injection. 3-NP up-regulated the expression of the autophagic protein beclin 1 but down-regulated the expression of the antiapoptotic protein Bcl-2. Pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) significantly inhibited the 3-NP-induced alterations in beclin 1 and Bcl-2 protein levels. Similarly, the 3-NP-induced decline in Bcl-2 was also prevented by the lysosomal inhibitor E64, indicating degradation of Bcl-2 by lysosomes. In agreement with the time course of 3-NP-induced cell death, an increase in the release of cytochrome c from mitochondria was observed. 3-MA also attenuated the 3-NP-induced release of cytochrome c. On the other hand, 3-NP-induced elevations in proapoptotic protein Bax and autophagic protein beclin 1 and LC3-II were significantly enhanced by the Bcl-2-specific inhibitor HA14-1. Furthermore, HA14-1 increased the release of cytochrome c and 3-NP-induced striatal damage. These results suggest that induction of autophagy leads to degradation of Bcl-2. Meanwhile, down-regulation of Bcl-2 amplifies autophagy activation and apoptotic signaling. Bcl-2 thus plays important roles in mitochondria dysfunction-induced apoptotic death of stritatal neurons by modulating both autophagic and apoptotic processes. © 2009 Wiley-Liss, Inc.

Huntington's disease (HD) is caused by an expansion of polyglutamine tract in N-terminus of huntingtin (htt). The mutation of htt leads to dysfunction and premature death of striatal and cortical neurons. However, the effects of htt mutation on glia remain largely unknown. This study aimed to establish a glia HD model using an adenoviral vector to express wild-type and mutant N-terminal huntingtin fragment 1-552 amino acids (htt552) in rat primary cortical astrocytes. We have evaluated optimal conditions for the infection of astrocytes with adenoviral vectors, and the kinetics of the expression of htt552 in astrocytes. The majority of astrocytes expressed the transgene after infection. At 24 h postinfection, the highest rate of infection was 89+/-3% for the wild-type (htt552-18Q) with a multiplicity of infection (m.o.i.) of 80, and the highest rate of infection was 91+/-4% for the mutant type (htt552-100Q) with the same viral dose. The duration of expression of htt552 lasted for about 7 days with a relatively high level from 1 to 4 days post-infection. Mutant huntingtin (htt552-100Q) produced the characteristic HD pathology after 3 days by the appearance of cytoplasmic aggregates and intranuclear inclusions. The result of MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide] assay showed that the inhibition of viability by virus on astrocytes was also dose-dependent. To obtain high infection rate and low toxicity, the viral dose with an m.o.i. of 40 was optimal to our cell model. The present study demonstrates that adenoviral-mediated expression of mutant htt provides an advantageous system for histological and biochemical analysis of HD pathogenesis in primary cortical astrocyte cultures.