Dl-3n-butylphthalide improves traumatic brain injury recovery via inhibiting autophagy-induced blood-brain barrier disruption and cell apoptosis.

Blood-brain barrier (BBB) disruption and neuronal apoptosis are important pathophysiological processes after traumatic brain injury (TBI). In clinical stroke, Dl-3n-butylphthalide (Dl-NBP) has a neuroprotective effect with anti-inflammatory, anti-oxidative, anti-apoptotic and mitochondrion-protective functions. However, the effect and molecular mechanism of Dl-NBP for TBI need to be further investigated. Here, we had used an animal model of TBI and SH-SY5Y/human brain microvascular endothelial cells to explore it. We found that Dl-NBP administration exerts a neuroprotective effect in TBI/OGD and BBB disorder, which up-regulates the expression of tight junction proteins and promotes neuronal survival via inhibiting mitochondrial apoptosis. The expressions of autophagy-related proteins, including ATG7, Beclin1 and LC3II, were significantly increased after TBI/OGD, and which were reversed by Dl-NBP treatment both in vivo and in vitro. Moreover, rapamycin treatment had abolished the effect of Dl-NBP for TBI recovery. Collectively, our current studies indicate that Dl-NBP treatment improved locomotor functional recovery after TBI by inhibiting the activation of autophagy and consequently blocking the junction protein loss and neuronal apoptosis. Dl-NBP, as an anti-inflammatory and anti-oxidative drug, may act as an effective strategy for TBI recovery.

Berberine attenuated the cytotoxicity induced by t-BHP via inhibiting oxidative stress and mitochondria dysfunction in PC-12 cells.

Neurodegenerative diseases all share several common features such as involvement of oxidative damage and mitochondrial dysfunction in pathogenesis. Oxidative stress induced by overproduction of mitochondrial reactive oxygen species (ROS) or impairment of the antioxidant deficiency results in mitochondrial dysfunction and initiation of the cell death cascade. Berberine (BBR), a traditional Chinese medicine, has been reported to exert anti-oxidative stress and anti-apoptosis effect in CNS diseases. However, the mechanism of BBR on regulating mitophagy and protecting mitochondrial function under oxidative stress remains unclear. In present study, we evaluated the beneficial effects of BBR on the tert-butyl hydroperoxide (t-BHP)-induced cytotoxicity. Furthermore, we explored the protective role of BBR in mitochondrial function and mitophagy under oxidative stress in PC-12 cells. Our results demonstrated that BBR effectively inhibited t-BHP-induced apoptosis which is associated with the decreased leakage of lactate dehydrogenase (LDH) and ROS overproduction. Moreover, BBR significantly suppressed cytochrome c expression, upregulated the ratio of Bcl-2/Bax, and ameliorated mitochondrial dysfunction by optimizing mitochondria membrane potential (ΔΨm) status and ATP production. In addition, BBR reduced the expression of autophagy-specific marker LC3, SQTM1/p62, and maintained lysosome normal function which involved the restoration of upstream signaling pathway AKT and mTOR phosphorylation level. Collectively, these findings suggested that BBR protects PC-12 cells from oxidative injury through inhibiting ROS level, mitochondria dysfunction, and mitophagy via PI3K/AKT/mTOR signaling pathways, which suggest a potential therapeutic strategy for oxidative stress and neurotoxic damages.

Valproate Attenuates Endoplasmic Reticulum Stress-Induced Apoptosis in SH-SY5Y Cells via the AKT/GSK3ß Signaling Pathway.

Endoplasmic reticulum (ER) stress-induced apoptosis plays an important role in a range of neurological disorders, such as neurodegenerative diseases, spinal cord injury, and diabetic neuropathy. Valproate (VPA), a typical antiepileptic drug, is commonly used in the treatment of bipolar disorder and epilepsy. Recently, VPA has been reported to exert neurotrophic effects and promote neurite outgrowth, but its molecular mechanism is still unclear. In the present study, we investigated whether VPA inhibited ER stress and promoted neuroprotection and neuronal restoration in SH-SY5Y cells and in primary rat cortical neurons, respectively, upon exposure to thapsigargin (TG). In SH-SY5Y cells, cell viability was detected by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, and the expression of ER stress-related apoptotic proteins such as glucose­regulated protein (GRP78), C/EBP homologous protein (CHOP), and cleaved caspase-12/-3 were analyzed with Western blot analyses and immunofluorescence assays. To explore the pathway involved in VPA-induced cell proliferation, we also examined p-AKT, GSK3ß, p-JNK and MMP-9. Moreover, to detect the effect of VPA in primary cortical neurons, immunofluorescence staining of ß-III tubulin and Anti-NeuN was analyzed in primary cultured neurons exposed to TG. Our results demonstrated that VPA administration improved cell viability in cells exposed to TG. In addition, VPA increased the levels of GRP78 and p-AKT and decreased the levels of ATF6, XBP-1, GSK3ß, p-JNK and MMP-9. Furthermore, the levels of the ER stress-induced apoptosis response proteins CHOP, cleaved caspase-12 and cleaved caspase-3 were inhibited by VPA treatment. Meanwhile, VPA administration also increased the ratio of Bcl-2/Bax. Moreover, VPA can maintain neurite outgrowth of primary cortical neurons. Collectively, the neurotrophic effect of VPA is related to the inhibition of ER stress-induced apoptosis in SH-SY5Y cells and the maintenance of neuronal growth. Collectively, our results suggested a new approach for the therapeutic function of VPA in neurological disorders and neuroprotection.

The Role of Histone H2B Acetylation Modification in Aluminum-Induced Cognitive Dysfunction.

Aluminum (Al) is a low toxic trace element that can accumulate in the nervous system and induce cognitive disorders characterized by reduced learning and memory ability. Neuroepigenetic effects are structural changes in cellular function by the brain in response to environmental stimuli by altering the expression of specific genes and repressing normal cellular transcription, leading to abnormalities in a variety of biological processes within the nervous system and affecting neurobehavioral responses. One of the most important mechanisms of epigenetic control on chromatin shape is histone modification. In the present study, we established an offspring rat model of Al intoxication to investigate the changes in spatial learning and memory retention abilities and the relationship with histone H2B acetylation modification in rats exposed to different doses of Al over a long period of time. The results demonstrated that long-term AlCl3 staining resulted in decreased CBP gene and protein expression, increased HDAC3 gene and protein levels, as well as decreased histone H2B and acH2BK20 protein expression levels in the hippocampus of rats. In conclusion, long-term exposure to Al may vary the expression of histone H2B and acH2BK20 through the regulation of enzymes that specifically regulate histone acetylation, hence hastening the deterioration of the nervous system that impairs cognitive function.

A Comparison of Endoscopic Closure and Laparoscopic Repair for Gastric Wall Defection.

Objective: To compare the effectiveness and safety of endoscopic closure and laparoscopic repair for gastric wall defection. Method: The clinical data of 120 patients with submucosal tumours enrolled at our hospital between January 2014 and December 2019 were retrospectively analysed. Patients were divided into two groups according to the surgery they underwent: an endoscopic closure group (n = 60) and a laparoscopic repair group (n = 60). The clinical characteristics, perioperative complications, and postoperative follow-up results of the two groups were analysed. Results: The surgery time in the endoscopic closure group was 56.20 ± 11.25 minutes, which was significantly lower compared with that in the laparoscopic repair group (159.35 ± 23.18 minutes; P < 0.001). In addition, the postoperative stay in the endoscopic closure group was shorter than that in the laparoscopic repair group, and the intraoperative bleeding volume and incidence of enteral nutrition initiation after surgery were significantly lower. Medical expenses were also significantly lower in the endoscopic closure group than in the laparoscopic repair group (P < 0.001). Only one patient developed a postoperative fever in the endoscopic closure group; three patients developed a postoperative fever and one patient had postoperative bleeding in the laparoscopic repair group. However, there were no statistical differences between the two groups regarding the incidence of R0 resection, postoperative fever, postoperative bleeding, and closure failure (all P > 0.05). There were no local recurrences, distant metastases, or deaths in either of the groups during the two-year follow-up period. Conclusion: Non-laparoscopic-assisted surgery may be quicker, safer, and more effective for gastric wall defection.

Praeruptorin B inhibits osteoclastogenesis by targeting GSTP1 and impacting on the S-glutathionylation of IKKß.

Osteoporosis a common disease in postmenopausal women which contains significant impact on the living quality of women. With the aging of the population, the number of patients suffer from osteoporosis has shown a significant increase. Given the limitations of clinical drugs for the treatment of osteoporosis, natural extracts with small side effects have a great application prospect in the treatment of osteoporosis. Praeruptorin B (Pra-B), is one of the main components found in the roots of Peucedanum praeruptorum Dunn and exhibits anti-inflammatory effects. However, there is no research on the influence of Pra-B on osteoporosis. Here, we showed that Pra-B can dose-dependently suppress osteoclastogenesis without cytotoxicity. Receptor activator of nuclear factor kappa-B (NF-κB) ligand (RANKL)-induced the nuclear import of P65 was inhibited by Pra-B, which indicated the suppressive effect of Pra-B on NF-κB signaling. Further, Pra-B enhanced the expression of Glutathione S-transferase Pi 1 (GSTP1) and promoted the S-glutathionylation of IKKß to inhibit the nuclear translocation of P65. Moreover, in vivo experiments showed that Pra-B considerably attenuated the bone loss in ovariectomy (OVX)-induced mice. Collectively, our studies revealed that Pra-B suppress the NF-κB signaling targeting GSTP1 to rescued RANKL-induced osteoclastogenesis in vitro and OVX-induced bone loss in vivo, supporting the potential of Pra-B for treating osteoporosis in the future.

Potentiometric Phosphate Ion Sensor Based on Electrochemical Modified Tungsten Electrode.

Determination of phosphate ions in aqueous solutions attracts a great deal of interest in the areas of environment, medicine, and agriculture. As phosphoric acid is a poly basic acid, the different forms of existence at different pH result in direct determination facing a big challenge. Herein, we reported a potentiometric phosphate ion sensor based on a surface-modified tungsten electrode. Pure tungsten was electrodeposited at a constant potential of 0.2 V versus Ag|AgCl in Na2HPO4. WO3 and H3O40PW12·xH2O were electrodeposited on the surface of the tungsten electrode. The modified tungsten electrode was used as a working electrode in a two-electrode system to detect the concentration of phosphate ions in aqueous solutions. The detection limit of the modified tungsten electrode for phosphate ions is 10-6 M from pH 7 to pH 8 and 10-5 M from pH 9 to pH 10. It has good selectivity to other common anions. The long-term monitoring experiment showed that the potential fluctuation was less than ±3 mV in 24 h. Compared to conventional determination methods, the current phosphate ion sensor showed a close value in a real sample. The mechanism of phosphate ion response was investigated in detail. This sensor possesses advantages of simple manufacture, low cost, a wide pH range for detecting, and good selectivity.

Automatic Management of Nutrient Solution for Hydroponics-Construction of Multi-ion Stat.

In order to improve plant factories, an appropriate control system on fertilization is urgently required. An automatic management system to control nutrient concentration was constructed using a programmable logic controller (PLC) and ion selective electrodes (ISEs) of nitrate, phosphate, and potassium ion. The concentration of nutrient components in a culture solution was monitored using these ISEs. When the concentration of the nutrient components diminished to the threshold set as an optimum condition (0.1 - 2.0 mM), an appropriate amount of a concentrated solution of each nutrient component was added to the culture solution using solenoid valves connected with the PLC. The present cultivation system was simply constructed without any computers and pumps. Three kinds of automatic control systems simultaneously worked and did not influence each other.

Fabrication of a Phosphate Ion Selective Electrode Based on Modified Molybdenum Metal.

A phosphate ion-selective electrode using molybdenum metal was constructed. The modified molybdenum electrode responded to HPO42- in the presence of molybdenum dioxide and molybdophosphate (PMo12O403-) on the surface. The electrode exhibited a linear response to HPO42- in the concentration range between 1.0 × 10-5 and 1.0 × 10-1 M (mol dm-3) in the pH range from 8.0 to 9.5 with a detection limit of 1.0 × 10-6 M. The sensor showed near Nernstian characteristics (27.8 ± 0.5 mV dec-1) at pH 9.0. Since the responding potential was attributed to the activity of HPO42-, the potential at a given concentration of phosphate depended on the pH. The electrode indicated a good selectivity with respect to other common anions such as NO3-, SO42-, Cl-, HCO3- and CH3COO-. The modified molybdenum electrode can be continuously used for over a 1 month with good reproducibility. The feasibility of the electrochemical sensor was proved by successful for the detection of phosphate in real samples.

Construction of an Automatic Nutrient Solution Management System for Hydroponics-Adjustment of the K+-Concentration and Volume of Water.

An automatic management system for nutrient solutions was constructed using a programmable logic controller (PLC) and a K+-ion selective electrode (K+-ISE). The concentration of K+ was monitored by the K+-ISE. When the concentration of K+ fell to the threshold limit, an appropriate amount of a concentrated K+ solution was added to the hydroponic solution. The volume was also maintained at a constant level by addition of water. This system can be constructed simply and inexpensively without any computers and pumps.

Hydrogen Sulfide Ameliorates Blood-Spinal Cord Barrier Disruption and Improves Functional Recovery by Inhibiting Endoplasmic Reticulum Stress-Dependent Autophagy.

Spinal cord injury (SCI) induces the disruption of blood-spinal cord barrier (BSCB), which elicits neurological deficits by triggering secondary injuries. Hydrogen sulfide (H2S) is a gaseous mediator that has been reported to have neuroprotective effect in the central nervous system. However, the relationship between H2S and BSCB disruption during SCI remains unknown. Therefore, it is interesting to evaluate whether the administration of NaHS, a H2S donor, can protect BSCB integrity against SCI and investigate the potential mechanisms underlying it. In present study, we found that SCI markedly activated endoplasmic reticulum (ER) stress and autophagy in a rat model of complete crushing injury to the spinal cord at T9 level. NaHS treatment prevented the loss of tight junction (TJ) and adherens junction (AJ) proteins both in vivo and in vitro. However, the protective effect of NaHS on BSCB restoration was significantly reduced by an ER stress activator (tunicamycin, TM) and an autophagy activator (rapamycin, Rapa). Moreover, SCI-induced autophagy was remarkably blocked by the ER stress inhibitor (4-phenylbutyric acid, 4-PBA). But the autophagy inhibitor (3-Methyladenine, 3-MA) only inhibited autophagy without obvious effects on ER stress. Finally, we had revealed that NaHS significantly alleviated BSCB permeability and improved functional recovery after SCI, and these effects were markedly reversed by TM and Rapa. In conclusion, our present study has demonstrated that NaHS treatment is beneficial for SCI recovery, indicating that H2S treatment is a potential therapeutic strategy for promoting SCI recovery.

Isoliquiritigenin protects against blood­brain barrier damage and inhibits the secretion of pro-inflammatory cytokines in mice after traumatic brain injury.

Traumatic brain injury (TBI) caused by an external mechanical force acting on the brain is a serious neurological condition. Inflammation plays an important role in prolonging secondary tissue injury after TBI, leading to neuronal cell death and dysfunction. Isoliquiritigenin (ILG) is a flavonoid monomer with anti-inflammatory characteristic. Thus, we had investigated the potential protective effects of ILG on TBI-induced injuries and identified the mechanisms underlying it. Here, we have demonstrated that ILG preserves blood brain barrier (BBB) integrity in vivo, suppresses the activation of microglia and inflammatory responses in mice after TBI, consequently leading to neurofunctional deficits, brain oedema, structural damage, and macrophage infiltration. In vitro, ILG exerts anti-inflammatory effect, and upregulates tight junction proteins 120­ß­catenin and occludin in SH­SY5Y cells under oxygen glucose deprivation/reoxygenation (OGD/D) condition. Additionally, we found that PI3K/AKT/GSK­3ß signalling pathway is involved in ILG treatment for TBI. To further confirm it, we had used SC79 (ethyl 2­amino­6­chloro­4­(1­cyano­2­ethoxy­2­oxoethyl)­4H­chromene­3­carboxylate), an Akt specific activator, to activate Akt, we found that SC79 partially reduces the protective effect of ILG for TBI. Overall, our current study reveals the neuroprotective role of ILG on TBI-induced BBB damage, downregulated tight junction proteins via PI3K/AKT/GSK­3ß signalling pathway. Furthermore, ILG suppresses the secretion of pro-inflammatory cytokines after TBI through inhibiting the PI3K/AKT/GSK­3ß/NF­κB signalling pathway. Our findings suggest that GSK­3ß is a key regulatory factor during TBI-induced secretion of inflammatory cytokines, neuronal apoptosis and destruction of BBB.

NaHS restores mitochondrial function and inhibits autophagy by activating the PI3K/Akt/mTOR signalling pathway to improve functional recovery after traumatic brain injury.

Traumatic brain injury (TBI) is one of the most serious public health problems in the world. TBI causes neurological deficits by triggering secondary injuries. Hydrogen sulfide (H2S), a gaseous mediator, has been reported to exert neuroprotective effects in central nervous system diseases, such as TBI. However, the molecular mechanisms involved in this effect are still unclear. The present study was designed to explore the ability of NaHS, a H2S donor, to provide neuroprotection in a mouse model of TBI and to discover the associated molecular mechanisms of these protective effects. Here, we found that administration of NaHS not only maintained the integrity of the blood brain barrier (BBB), protected neurons from apoptosis, and promoted remyelination and axonal reparation but also protected mitochondrial function. In addition, we found that autophagy was inhibited after treatment with NaHS following TBI, an effect that was induced by activation of the PI3K/AKT/mTOR signalling pathway. Our study indicated that H2S treatment is beneficial for TBI, pointing to H2S as a potential therapeutic target for treating TBI.

Melatonin protects against blood-brain barrier damage by inhibiting the TLR4/ NF-κB signaling pathway after LPS treatment in neonatal rats.

Hypoxic-ischemic and inflammatory (HII) induces the disruption of blood-brain barrier (BBB) which leads to inflammatory responses and neuronal cell death, resulting in brain secondary damage. Previous studies showed that melatonin produced potent neuroprotective effects in neonatal hypoxic-ischaemic models. However, the relationship between BBB disruption and melatonin in HII was still unclear. The present study therefore investigated the beneficial effects of melatonin on BBB after HII and the underlying mechanisms. HII animal model was conducted by receiving lipopolysaccharide followed by 90 min hypoxia-ischaemia in postnatal day 2 Sprague-Dawley rat pups. Melatonin was injected intraperitoneally 1 h before lipopolysaccharide injection and then once a day for 1 week to evaluate the long-term effects. In this study, we demonstrated that melatonin administration inhibited the disruption of BBB permeability and improved the white matter recovery in HII model rats. Melatonin significantly attenuated the degradation of junction proteins and the neuroprotective role was related to the inhibition of microglial toll-like receptor 4/ nuclear factor-kappa B signaling pathway both in vivo and in vitro. Taken together, our data demonstrated that therapeutic strategies targeting inflammation might be suitable for the therapy of preserving BBB integrity after HII.

Resveratrol post-treatment protects against neonatal brain injury after hypoxia-ischemia.

Neonatal hypoxic-ischemic brain injury is a devastating disease with limited treatment options. Preventive treatment with resveratrol has indicated to be well tolerated and has lower toxicity in both experimental models and human patients. However, whether resveratrol administration post-hypoxic-ischemic protects against neonatal hypoxic-ischemic injury is not known. Here we reported that post-treatment with resveratrol significantly reduced brain damage at 7-day after the injury. We found that resveratrol reduced the expression levels of key inflammatory factors at the mRNA and protein levels, and at least partially via inhibiting microglia activation. Moreover, resveratrol exerted an anti-apoptotic effect, as assessed by TUNEL staining, and altered the expression of the apoptosis-related genes Bax, Bcl-2 and caspase3. Our data indicate that post-treatment with resveratrol protects against neonatal hypoxic-ischemic brain injury and suggest a promising therapeutic strategy to this disease.

Learning nonlinear multiregression networks based on evolutionary computation.

This paper describes a novel knowledge discovery and data mining framework dealing with nonlinear interactions among domain attributes. Our network-based model provides an effective and efficient reasoning procedure to perform prediction and decision making. Unlike many existing paradigms based on linear models, the attribute relationship in our framework is represented by nonlinear nonnegative multiregressions based on the Choquet integral. This kind of multiregression is able to model a rich set of nonlinear interactions directly. Our framework involves two layers. The outer layer is a network structure consisting of network elements as its components, while the inner layer is concerned with a particular network element modeled by Choquet integrals. We develop a fast double optimization algorithm (FDOA) for learning the multiregression coefficients of a single network element. Using this local learning component and multiregression-residual-cost evolutionary programming (MRCEP), we propose a global learning algorithm, called MRCEP-FDOA, for discovering the network structures and their elements from databases. We have conducted a series of experiments to assess the effectiveness of our algorithm and investigate the performance under different parameter combinations, as well as sizes of the training data sets. The empirical results demonstrate that our framework can successfully discover the target network structure and the regression coefficients.

Effect of route of preoperative biopsy on endoscopic submucosal dissection for patients with early gastric cancer.

OBJECTIVE: To observe and compare the effects of multi-patch biopsy under conventional white light imaging endoscopy (C-WLI) and precise targeted biopsy under magnifying narrow-band imaging endoscopy (M-NBI) on the endoscopic submucosal dissection (ESD) of early gastric cancers and intraepithelial neoplasias. METHODS: According to the way of selecting biopsy specimens, patients were divided into C-WLI and M-NBI groups, 20 cases. The ESD operations of the 2 groups were compared quantitively. RESULTS: The mean frequency of biopsy in M-NBI group was (1.00±0.00), obviously lower than in the C-WLI group (4.78±1.02) (P<0.01).The average total number of selected biopsy specimens was also fewer (1.45±0.12 and 7.82±2.22, respectively, P<0.01). There was no significant difference in the time of determining excision extension, marking time and the time of specimen excision of 2 groups during the ESD (P>0.05), whereas submucosal injection time, mucosal dissection time, stopping bleeding time, wound processing time in the M-NBI group were significantly shorter than in the C-WLI group (P<0.01). CONCLUSION: Precise targeted biopsy under M-NBI can obviously shorten the time of ESD operation, with small quantity of tissues but high pathological positive rate.