Roflupram alleviates autophagy defects and reduces mutant hSOD1-induced motor neuron damage in cell and mouse models of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal and incurable disease involving motor neuron (MN) degeneration and is characterized by ongoing myasthenia and amyotrophia in adults. Most ALS patients die of respiratory muscle paralysis after an average of 3-5 years. Defective autophagy in MNs is considered an important trigger of ALS pathogenesis. Roflupram (ROF) was demonstrated to activate autophagy in microglial cells and exert protective effects against Parkinson's disease (PD) and Alzheimer's disease (AD). Therefore, our research aimed to investigate the efficacy and mechanism of ROF in treating ALS both in vivo and in vitro. We found that ROF could delay disease onset and prolong the survival of hSOD1-G93A transgenic mice. Moreover, ROF protected MNs in the anterior horn of the spinal cord, activated the AMPK/ULK1 signaling pathway, increased autophagic flow, and reduced SOD1 aggregation. In an NSC34 cell line stably transfected with hSOD1-G93A, ROF protected against cellular damage caused by hSOD1-G93A. Moreover, we have demonstrated that ROF inhibited gliosis in ALS model mice. Collectively, our study suggested that ROF is neuroprotective in ALS models and the AMPK/ULK1 signaling pathway is a potential therapeutic target in ALS, which increases autophagic flow and reduces SOD1 aggregation.

Researches of calcium-activated chloride channel ANO1 intervening amyotrophic lateral sclerosis progression by activating EGFR and CaMKII signaling.

BACKGROUND: ANO1 is closely correlated with the activation of EGFR and CaMKII, while EGFR and CaMKII show low activation in amyotrophic lateral sclerosis (ALS) models. Therefore, we designed experiments to verify that ANO1 may play a protective role on motor neurons in ALS by activating EGFR and CaMKII. METHODS: The expression changes of ANO1, EGFR, CaMKII, pEGFR, and pCaMKII, cell survival status, and apoptosis were studied by western blot, real-time quantitative PCR, immunofluorescence, immunohistochemistry, CCK-8, and flow cytometry. The role of ANO1 in the ALS model by activating EGFR and CaMKII was studied by applying corresponding activators, inhibitors, gene silencing, and overexpression. RESULTS: In hSOD1G93A transgenic animals and cell lines, low expression of ANO1 and low activation of EGFR and CaMKII were identified. ANO1 expression decreased gradually with the progression of ALS. Overexpression of ANO1 in the hSOD1G93A cell line and primary neurons of hSOD1G93A transgenic mice increased cell viability and decreased cell apoptosis. After the application of ANO1 inhibitor CaCC-inhA01 in hSOD1G93A cell line and primary neurons of hSOD1G93A transgenic mice, EGFR activator EGF and CaMKII activator Carbachol, increased cell viability and reduced cell apoptosis. After ANO1 was overexpressed in the hSOD1G93A cell line and primary neurons of hSOD1G93A transgenic mice, EGFR inhibitor AEE788 and CaMKII inhibitor KN93 decreased cell viability and increased cell apoptosis. CONCLUSIONS: Our results suggest that ANO1 plays an important role in the survival of ALS motor neurons. ANO1 can increase cell activity and reduce apoptosis by activating EGFR and CaMKII signals.

SPY1 inhibits neuronal ferroptosis in amyotrophic lateral sclerosis by reducing lipid peroxidation through regulation of GCH1 and TFR1.

Ferroptosis is an iron-dependent cell death with the accumulation of lipid peroxidation and dysfunction of antioxidant systems. As the critical regulator, glutathione peroxidase 4 (GPX4) has been demonstrated to be down-regulated in amyotrophic lateral sclerosis (ALS). However, the mechanism of ferroptosis in ALS remains unclear. In this research, bioinformatics analysis revealed a high correlation between ALS, ferroptosis, and Speedy/RINGO cell cycle regulator family member A (SPY1). Lipid peroxidation of ferroptosis in hSOD1G93A cells and mice was generated by TFR1-imported excess free iron, decreased GSH, mitochondrial membrane dysfunction, upregulated ALOX15, and inactivation of GCH1, GPX4. SPY1 is a "cyclin-like" protein that has been proved to enhance the viability of hSOD1G93A cells by inhibiting DNA damage. In our study, the decreased expression of SPY1 in ALS was resulted from unprecedented ubiquitination degradation mediated by MDM2 (a nuclear-localized E3 ubiquitin ligase). Further, SPY1 was identified as a novel ferroptosis suppressor via alleviating lipid peroxidation produced by dysregulated GCH1/BH4 axis (a resistance axis of ferroptosis) and transferrin receptor protein 1 (TFR1)-induced iron. Additionally, neuron-specific overexpression of SPY1 significantly delayed the occurrence and prolonged the survival in ALS transgenic mice through the above two pathways. These results suggest that SPY1 is a novel target for both ferroptosis and ALS.

Adjunctive accelerated repetitive transcranial magnetic stimulation for older patients with depression: A systematic review.

Objective: We performed this systemic review to investigate the therapeutic potential and safety of adjunctive accelerated repetitive transcranial magnetic stimulation (aTMS) for older patients with depression. Methods: We included published randomized clinical trials (RCTs) and observational studies targeting adjunctive aTMS for older patients with depression. Results: Two open-label self-controlled studies (n = 29) fulfilled the criteria for inclusion. The included studies reported significant improvements in depressive symptoms from baseline to post-aTMS (all Ps < 0.05). One study reported a dropout rate of 10.5% (2/19). Mild headache was the most common adverse reaction. Conclusion: The currently available evidence from two open-label self-controlled studies indicates that adjunctive aTMS is a safe and effective therapy for older patients with depression.

[Vertical Distribution Characteristics and Influencing Factors of Phytoplankton Community Structure in Qiandao Lake].

Clear vertical variations in phytoplankton community structure are usually observed in deep-water lakes and reservoirs, which is one of the key components of water quality and ecosystem functioning. However, the vertical patterns and ecological drivers of phytoplankton communities in deep-water lakes and reservoirs are still understudied. In this study, we took Qiandao Lake, a deep-water reservoir, as an example to reveal the vertical distribution characteristics of phytoplankton communities and its influencing factors by investigating phytoplankton community structure and the associated water quality index at 12 sites across the whole lake in two seasons (spring and autumn). The results showed that the phytoplankton abundance and chlorophyll a were highest in the surface layer in autumn and then decreased toward deep water, whereas in spring, the maximum value occurred in the subsurface layer (2-5 m), and the dominant phytoplankton species showed obvious vertical stratification characteristics. Specifically, in spring, Cryptomonas and Pseudanabaena dominated the surface and subsurface layers, Cryptomonas dominated in the middle layer, and the abundance of Cyclotella at the bottom layer was significantly higher than that of the other algae genera. The dominant genera in autumn were Pseudanabaena and Aphanizomenon. In the subsurface and middle layers, Leptolyngbya and Pseudanabaena occupied the dominant position, and Leptolyngbya became the only dominant genus. In the bottom layer, Leptolyngbya was the only dominant genus. The key environmental indicators of the water also had obvious vertical changes. The contents of N and P nutrients had a negative correlation with the water depth in spring, whereas the reverse trend was observed in autumn. The correlation analysis showed that the vertical variation in phytoplankton abundance in spring was significantly positively correlated with phosphate concentration, whereas the vertical distribution of phytoplankton abundance in autumn was significantly positively correlated with intensity of light, and the water temperature, NH4+-N, and total nitrogen were the main factors driving the vertical changes in the dominant genera of phytoplankton community in the two seasons. To summarize, environmental conditions such as water temperature, light, and nutrients had strong effects on the vertical distribution of phytoplankton. In the ecological investigation and quality assessment of deep-water lakes and reservoirs, the vertical distribution characteristics of the phytoplankton community structure and the influence of environmental conditions should be fully considered.

Hydrothermal synthesis of Bi2Se3 nanosheets by using gallic acid as a reductant.

In this work, a simple and reproducible hydrothermal synthesis was employed to synthesize two-dimensional Bi2Se3 nanosheets by using gallic acid as a reductant. Meanwhile, the effects of the amounts of gallic acid and sodium hydroxide and the surfactant Triton X-100 on phase composition and morphology of the obtained Bi2Se3 were also studied. The results reveal that gallic acid could effectively reduce Se4+ to Se2- and gave rise to the formation of Bi2Se3. Additionally, keeping the reaction conditions of molar ratio of gallic acid to the precursor elements (Bi + Se) at 1 to 1 (or higher) and using strong alkaline solutions were the key factors to synthesize high purity crystalline Bi2Se3 nanosheets. Furthermore, flower-like Bi2Se3 composed of nanosheets with a dozen nanometer thickness could be easily fabricated by adding appropriate amounts of Triton X-100. This work provides a novel approach for synthesis of ultra-thin Bi2Se3 nanosheets in a controllable manner.

PAK4 suppresses motor neuron degeneration in hSOD1G93A -linked amyotrophic lateral sclerosis cell and rat models.

OBJECTIVES: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neurons (MN). CREB pathway-mediated inhibition of apoptosis contributes to neuron protection, and PAK4 activates CREB signalling in diverse cell types. This study aimed to investigate PAK4's effect and mechanism of action in ALS. METHODS: We analysed RNA levels by qRT-PCR, protein levels by immunofluorescence and Western blotting, and apoptosis by flow cytometry and TUNEL staining. Cell transfection was performed for in vitro experiment. Mice were injected intraspinally to evaluate PAK4 function in vivo experiment. Rotarod test was performed to measure motor function. RESULTS: The expression and activation of PAK4 significantly decreased in the cell and mouse models of ALS as the disease progressed, which was caused by the negative regulation of miR-9-5p. Silencing of PAK4 increased the apoptosis of MN by inhibiting CREB-mediated neuroprotection, whereas overexpression of PAK4 protected MN from hSOD1G93A -induced degeneration by activating CREB signalling. The neuroprotective effect of PAK4 was markedly inhibited by CREB inhibitor. In ALS models, the PAK4/CREB pathway was inhibited, and cell apoptosis increased. In vivo experiments revealed that PAK4 overexpression in the spinal neurons of hSOD1G93A mice suppressed MN degeneration, prolonged survival and promoted the CREB pathway. CONCLUSIONS: PAK4 protects MN from degeneration by activating the anti-apoptotic effects of CREB signalling, suggesting it may be a therapeutic target in ALS.

Diphenyl diselenide protects motor neurons through inhibition of microglia-mediated inflammatory injury in amyotrophic lateral sclerosis.

Microglia-mediated neuroinflammatory response and neuron damage are considered as a self-propelling progressive cycle, being strongly implicated in the progression of neurodegeneration in amyotrophic lateral sclerosis (ALS). Diphenyl diselenide (DPDS), a simple organoselenium compound, has been known to possess multiple pharmacological properties. The purpose of this study was to explore the neuroprotective effects of DPDS against microglia-mediated neuroinflammatory injury in ALS models. We found that DPDS pretreatment inhibited LPS-induced activation of IκB/NF-κB pathway and subsequent release of proinflammatory factors from activated primary hSOD1G93A microglia. Moreover, DPDS suppressed NLRP3 inflammasome activation by decreasing protein nitration via reduction in NO and ROS levels, whose low levels are related to NF-κB inhibition responsible for iNOS and NOX2 down-regulations, respectively. Notably, DPDS-mediated ROS attenuation was not linked to Nrf2 activation in this cellular model. Furthermore, in the absence of activated microglia, DPDS has no significant effect on the individual hSOD1G93A-NSC34 cells; however, in in vitro neuron-microglia conditional culture and co-culture experiments, DPDS protected motor neurons from neurotoxic damage caused by LPS or BzATP-stimulated microglia activation. Above observations suggest that DPDS-afforded neuroprotection is linked to inhibition of microglia-mediated neuroinflammation in ALS, which was further verified in vivo as shown by improvements of motor deficits, prolonged survival, and reduction of motor neuron loss and reactive microgliosis in hSOD1G93A transgenic mouse. Altogether, our results show that DPDS elicited neuroprotection in ALS models through inactivation of microglia by inhibiting IκB/NF-κB pathway and NLRP3 inflammasome activation, suggesting that DPDS may be a promising candidate for potential therapy for ALS.

Adjunctive Magnetic Seizure Therapy for Schizophrenia: A Systematic Review.

Objective: The efficacy and safety of adjunctive magnetic seizure therapy (MST) for patients with schizophrenia are unclear. This systematic review was conducted to examine the efficacy and safety of adjunctive MST for schizophrenia. Methods: Chinese (WanFang and Chinese Journal Net) and English (PubMed, EMBASE, PsycINFO, and the Cochrane Library) databases were systematically searched. Results: Two open-label self-controlled studies (n = 16) were included and analyzed in this review. In these studies, the Positive and Negative Syndrome Scale (PANSS) total scores and Brief Psychiatric Rating Scale (BPRS) total scores significantly decreased from baseline to post-MST (all Ps < 0.05), without serious adverse neurocognitive effects. Mixed findings on the neurocognitive effects of adjunctive MST for schizophrenia were reported in the two studies. A discontinuation rate of treatment of up to 50% (4/8) was reported in both studies. The rate of adverse drug reactions (ADRs) was evaluated in only one study, where the most common ADRs were found to be dizziness (25%, 2/8) and subjective memory loss (12.5%, 1/8). Conclusion: There is inconsistent evidence for MST-related adverse neurocognitive effects and preliminary evidence for the alleviation of psychotic symptoms in schizophrenia.

The impact of the COVID-19 pandemic on firms: a survey in Guangdong Province, China.

Background: The COVID-19 pandemic has affected all sectors of the world economy and society. To understand the impact of the pandemic on firms in China and suggest public policies to deal with the negative effects, we investigated firms in Guangdong Province. Methods: The survey sample included 524 firms in 15 cities in Guangdong Province. We chose these firms from the list published by the government, considering the industrial characteristics of Guangdong province and firm size. The questionnaire was developed based on previous studies carried out by UNDP. It comprised four categories with a total of 17 questions. The executives of firms were contacted by telephone or WeChat, and were invited to answer self-administered questionnaires through an online survey platform. The data was analyzed by SPSS. Results: The following key findings were revealed through the survey: (1) 48.7% of firms maintained stability, and 35.1% experienced a halt in operation or faced closure; (2) Nearly 70-90% already exploit online commerce or are willing to do so, and also remote office work, and digital operations. (3) 46% believe that they will certainly incur losses for 2020, and 83.5% expect the city's GDP to decrease. Conclusions: Firms in Guangdong Province have faced great challenges in the epidemic. Their production and operation activities have been limited, and they are facing significant risks. It is necessary to implement policies that would profoundly lower production costs for firms, help them survive this difficult period, and gradually return to normal business.

Identification of potential drugs for Parkinson's disease based on a sub-pathway method.

Parkinson's disease (PD) is the second most prevalent neurodegenerative disease in ageing individuals. Current therapeutic regimen suffers from general side effects and a poor efficiency for PD symptoms. The need for development new therapeutic agents for PD is urgent. Here, we aimed to explore the metabolic mechanism of PD and identified potential novel agents for PD by a sub-pathway-based method. By using the GSE7621 microarray data from the GEO database, we first identified the 1226 differentially expressed genes (DEGs) between PD and normal samples. Then we identified 19 significant enriched metabolic sub-pathways, which may involve in development of PD. Finally, by an integrated analysis of PD-involved sub-pathways and drug-affected sub-pathways, we identified 49 novel small molecular drugs capable to target the PD-involved sub-pathways. Our method could not only identify existing drug (apomorphine) for PD, but also predict potentially novel agents (ketoconazole and astemizole), which might have therapeutic effects via targeting some key enzymes in arachidonic acid metabolism. These candidate agents identified by our approach may provide insights into a novel therapy approach for PD.

Combined flame and solution synthesis of nanoscale tungsten-oxide and zinc/tin-oxide heterostructures.

Heterostructures of tungsten-oxide nanowires decorated with zinc/tin-oxide nanostructures are synthesized via a combined flame and solution synthesis approach. Vertically well-aligned tungsten-oxide nanowires are grown on a tungsten substrate by a flame synthesis method. Here, tetragonal WO(2.9) nanowires (diameters of 20-50 nm, lengths >10 µm, and coverage density of 10(9)-10(10) cm(-2)) are produced by the vapor-solid mechanism at 1720 K. Various kinds of Zn/Sn-oxide nanostructures are grown or deposited on the WO(2.9) nanowires by adjusting the Sn(2+) : Zn(2+) molar ratio in an aqueous ethylenediamine solution at 65 °C. With WO(2.9) nanowires serving as the base structures, sequential growth or deposition on them of hexagonal ZnO nanoplates, Zn(2)SnO(4) nanocubes, and SnO(2) nanoparticles are attained for Sn(2+) : Zn(2+) ratios of 0 : 1, 1 : 10, and 10 : 1, respectively, along with different saturation conditions. High-resolution transmission electron microscopy of the interfaces at the nanoheterojunctions shows abrupt interfaces for ZnO/WO(2.9) and Zn(2)SnO(4)/WO(2.9), despite lattice mismatches of >20%.

[Protective effect of Yigan Fuzheng Paidu capsules combined with ozone on CCl4-induced acute hepatic injury in dogs].

OBJECTIVE: To investigate the protective effect of Yigan Fuzheng Paidu Capsules (YC) combined with medical ozone against hepatic injury in dogs induced by hepatotoxic drug. METHODS: Twenty-four dogs were randomized equally into 4 groups (n=6), namely the model group, oleanolic acid tablet (OAT) group, YC group and YC+O(3) group, given either no particular treatment, oral OAT at 10 mg/day, oral YC at 0.2 g/day, or YC at 0.2 g/day plus 150 ml medical ozone transrectal insufflation every other day, respectively, for totally 30 consecutive days. Acute hepatic injury was induced after the treatment in the dogs with a sing-dose intraperitoneal injection of 0.9 ml/kg CCl(4) and peanut oil mixture (1:1, W/W). The general condition, survival time, alanine aminotransferase (ALT), aspartate aminotransferase/alanine aminotransferase (AST/ALT), serum total bilirubin (TBIL), prothrombin time (PT), blood ammonia (AMMO), and blood urea nitrogen (BUN) were recorded or measured. The hepatic pathological changes were observed upon death or on day 15 following CCl(4) injection. RESULTS: Compared with the other 3 treatment protocols, YC plus O(3) showed favorable effects on the activity, mental state, diet, urination and defecation of the dogs, which had significantly higher survival rate and higher levels of ALT, TBIL, PT, and AMMO than the model and OAT groups (P<0.05). AST/ALT remained normal in YC+O(3) group, which had also milder hepatic injury than the other 3 groups. CONCLUSIONS: YC combined with medical ozone may decrease transaminase and blood ammonia levels, relieve jaundice, prolong the survival time of dogs with CCl(4)-induced hepatic injury.