Ancient dormant virus remnant ERVW-1 drives ferroptosis via degradation of GPX4 and SLC3A2 in schizophrenia.

Human endogenous retroviruses (HERVs) are remnants of retroviral infections in human germline cells from millions of years ago. Among these, ERVW-1 (also known as HERV-W-ENV, ERVWE1, or ENVW) encodes the envelope protein of the HERV-W family, which contributes to the pathophysiology of schizophrenia. Additionally, neuropathological studies have revealed cell death and disruption of iron homeostasis in the brains of individuals with schizophrenia. Here, our bioinformatics analysis showed that differentially expressed genes in the human prefrontal cortex RNA microarray dataset (GSE53987) were mainly related to ferroptosis and its associated pathways. Clinical data demonstrated significantly lower expression levels of ferroptosis-related genes, particularly Glutathione peroxidase 4 (GPX4) and solute carrier family 3 member 2 (SLC3A2), in schizophrenia patients compared to normal controls. Further in-depth analyses revealed a significant negative correlation between ERVW-1 expression and the levels of GPX4/SLC3A2 in schizophrenia. Studies indicated that ERVW-1 increased iron levels, malondialdehyde (MDA), and transferrin receptor protein 1 (TFR1) expression while decreasing glutathione (GSH) levels and triggering the loss of mitochondrial membrane potential, suggesting that ERVW-1 can induce ferroptosis. Ongoing research has shown that ERVW-1 reduced the expression of GPX4 and SLC3A2 by inhibiting their promoter activities. Moreover, Ferrostatin-1 (Fer-1), the ferroptosis inhibitor, reversed the iron accumulation and mitochondrial membrane potential loss, as well as restored the expressions of ferroptosis markers GSH, MDA, and TFR1 induced by ERVW-1. In conclusion, ERVW-1 could promote ferroptosis by downregulating the expression of GPX4 and SLC3A2, revealing a novel mechanism by which ERVW-1 contributes to neuronal cell death in schizophrenia.

Captive ERVWE1 triggers impairment of 5-HT neuronal plasticity in the first-episode schizophrenia by post-transcriptional activation of HTR1B in ALKBH5-m6A dependent epigenetic mechanisms.

BACKGROUND: Abnormalities in the 5-HT system and synaptic plasticity are hallmark features of schizophrenia. Previous studies suggest that the human endogenous retrovirus W family envelope (ERVWE1) is an influential risk factor for schizophrenia and inversely correlates with 5-HT4 receptor in schizophrenia. To our knowledge, no data describes the effect of ERVWE1 on 5-HT neuronal plasticity. N6-methyladenosine (m6A) regulates gene expression and impacts synaptic plasticity. Our research aims to systematically investigate the effects of ERVWE1 on 5-HT neuronal plasticity through m6A modification in schizophrenia. RESULTS: HTR1B, ALKBH5, and Arc exhibited higher levels in individuals with first-episode schizophrenia compared to the controls and showed a strong positive correlation with ERVWE1. Interestingly, HTR1B was also correlated with ALKBH5 and Arc. Further analyses confirmed that ALKBH5 may be an independent risk factor for schizophrenia. In vitro studies, we discovered that ERVWE1 enhanced HTR1B expression, thereby activating the ERK-ELK1-Arc pathway and reducing the complexity and spine density of 5-HT neurons. Furthermore, ERVWE1 reduced m6A levels through ALKBH5 demethylation. ERVWE1 induced HTR1B upregulation by improving its mRNA stability in ALKBH5-m6A-dependent epigenetic mechanisms. Importantly, ALKBH5 mediated the observed alterations in 5-HT neuronal plasticity induced by ERVWE1. CONCLUSIONS: Overall, HTR1B, Arc, and ALKBH5 levels were increased in schizophrenia and positively associated with ERVWE1. Moreover, ALKBH5 was a novel risk gene for schizophrenia. ERVWE1 impaired 5-HT neuronal plasticity in ALKBH5-m6A dependent mechanism by the HTR1B-ERK-ELK1-Arc pathway, which may be an important contributor to aberrant synaptic plasticity in schizophrenia.

HERV-W ENV Induces Innate Immune Activation and Neuronal Apoptosis via linc01930/cGAS Axis in Recent-Onset Schizophrenia.

Schizophrenia is a severe neuropsychiatric disorder affecting about 1% of individuals worldwide. Increased innate immune activation and neuronal apoptosis are common findings in schizophrenia. Interferon beta (IFN-ß), an essential cytokine in promoting and regulating innate immune responses, causes neuronal apoptosis in vitro. However, the precise pathogenesis of schizophrenia is unknown. Recent studies indicate that a domesticated endogenous retroviral envelope glycoprotein of the W family (HERV-W ENV, also called ERVWE1 or syncytin 1), derived from the endogenous retrovirus group W member 1 (ERVWE1) locus on chromosome 7q21.2, has a high level in schizophrenia. Here, we found an increased serum IFN-ß level in schizophrenia and showed a positive correlation with HERV-W ENV. In addition, serum long intergenic non-protein coding RNA 1930 (linc01930), decreased in schizophrenia, was negatively correlated with HERV-W ENV and IFN-ß. In vitro experiments showed that linc01930, mainly in the nucleus and with noncoding functions, was repressed by HERV-W ENV through promoter activity suppression. Further studies indicated that HERV-W ENV increased IFN-ß expression and neuronal apoptosis by restraining the expression of linc01930. Furthermore, HERV-W ENV enhanced cyclic GMP-AMP synthase (cGAS) and stimulator of interferon genes protein (STING) expression and interferon regulatory factor 3 (IRF3) phosphorylation in neuronal cells. Notably, cGAS interacted with HERV-W ENV and triggered IFN-ß expression and neuronal apoptosis caused by HERV-W ENV. Moreover, Linc01930 participated in the increased neuronal apoptosis and expression level of cGAS and IFN-ß induced by HERV-W ENV. To summarize, our results suggested that linc01930 and IFN-ß might be novel potential blood-based biomarkers in schizophrenia. The totality of these results also showed that HERV-W ENV facilitated antiviral innate immune response, resulting in neuronal apoptosis through the linc01930/cGAS/STING pathway in schizophrenia. Due to its monoclonal antibody GNbAC1 application in clinical trials, we considered HERV-W ENV might be a reliable therapeutic choice for schizophrenia.

ERVWE1 Reduces Hippocampal Neuron Density and Impairs Dendritic Spine Morphology through Inhibiting Wnt/JNK Non-Canonical Pathway via miR-141-3p in Schizophrenia.

Human endogenous retroviruses (HERVs) are remnants of ancestral germline infections by exogenous retroviruses. Human endogenous retroviruses W family envelope gene (HERV-W env, also called ERVWE1), located on chromosome 7q21-22, encodes an envelope glycoprotein from the HERV-W family. Mounting evidence suggests that aberrant expression of ERVWE1 involves the etiology of schizophrenia. Moreover, the genetic and morphological studies indicate that dendritic spine deficits may contribute to the onset of schizophrenia. Here, we reported that ERVWE1 changed the density and morphology of the dendritic spine through inhibiting Wingless-type (Wnt)/c-Jun N-terminal kinases (JNK) non-canonical pathway via miR-141-3p in schizophrenia. In this paper, we found elevated levels of miR-141-3p and a significant positive correlation with ERVWE1 in schizophrenia. Moreover, serum Wnt5a and actin-related protein 2 (Arp2) levels decreased and demonstrated a significant negative correlation with ERVWE1 in schizophrenia. In vitro experiments disclosed that ERVWE1 up-regulated miR-141-3p expression by interacting with transcription factor (TF) Yin Yang 1 (YY1). YY1 modulated miR-141-3p expression by binding to its promoter. The luciferase assay revealed that YY1 enhanced the promoter activity of miR-141-3p. Using the miRNA target prediction databases and luciferase reporter assays, we demonstrated that miR-141-3p targeted Wnt5a at its 3' untranslated region (3' UTR). Furthermore, ERVWE1 suppressed the expression of Arp2 through non-canonical pathway, Wnt5a/JNK signaling pathway. In addition, ERVWE1 inhibited Wnt5a/JNK/Arp2 signal pathway through miR-141-3p. Finally, functional assays showed that ERVWE1 induced the abnormalities in hippocampal neuron morphology and spine density through inhibiting Wnt/JNK non-canonical pathway via miR-141-3p in schizophrenia. Our findings indicated that miR-141-3p, Wnt5a, and Arp2 might be potential clinical blood-based biomarkers or therapeutic targets for schizophrenia. Our work also provided new insight into the role of ERVWE1 in schizophrenia pathogenesis.

Domesticated HERV-W env contributes to the activation of the small conductance Ca2+-activated K+ type 2 channels via decreased 5-HT4 receptor in recent-onset schizophrenia.

The human endogenous retroviruses type W family envelope (HERV-W env) gene is located on chromosome 7q21-22. Our previous studies show that HERV-W env is elevated in schizophrenia and HERV-W env can increase calcium influx. Additionally, the 5-HTergic system and particularly 5-hydroxytryptamine (5-HT) receptors play a prominent role in the pathogenesis and treatment of schizophrenia. 5-hydroxytryptamine receptor 4 (5-HT4R) agonist can block calcium channels. However, the underlying relationship between HERV-W env and 5-HT4R in the etiology of schizophrenia has not been revealed. Here, we used enzyme-linked immunosorbent assay to detect the concentration of HERV-W env and 5-HT4R in the plasma of patients with schizophrenia and we found that there were decreased levels of 5-HT4R and a negative correlation between 5-HT4R and HERV-W env in schizophrenia. Overexpression of HERV-W env decreased the transcription and protein levels of 5-HT4R but increased small conductance Ca2+-activated K+ type 2 channels (SK2) expression levels. Further studies revealed that HERV-W env could interact with 5-HT4R. Additionally, luciferase assay showed that an essential region (-364 to -176 from the transcription start site) in the SK2 promoter was required for HERV-W env-induced SK2 expression. Importantly, 5-HT4R participated in the regulation of SK2 expression and promoter activity. Electrophysiological recordings suggested that HERV-W env could increase SK2 channel currents and the increase of SK2 currents was inhibited by 5-HT4R. In conclusion, HERV-W env could activate SK2 channels via decreased 5-HT4R, which might exhibit a novel mechanism for HERV-W env to influence neuronal activity in schizophrenia.

CPEB1, a novel risk gene in recent-onset schizophrenia, contributes to mitochondrial complex I defect caused by a defective provirus ERVWE1.

BACKGROUND: Schizophrenia afflicts 1% of the world population. Clinical studies suggest that schizophrenia patients may have an imbalance of mitochondrial energy metabolism via inhibition of mitochondrial complex I activity. Moreover, recent studies have shown that ERVWE1 is also a risk factor for schizophrenia. Nevertheless, there is no available literature concerning the relationship between complex I deficits and ERVWE1 in schizophrenia. Identifying risk factors and blood-based biomarkers for schizophrenia may provide new guidelines for early interventions and prevention programs. AIM: To address novel potential risk factors and the underlying mechanisms of mitochondrial complex I deficiency caused by ERVWE1 in schizophrenia. METHODS: Quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay were used to detect differentially expressed risk factors in blood samples. Clinical statistical analyses were performed by median analyses and Mann-Whitney U analyses. Spearman's rank correlation was applied to examine the correlation between different risk factors in blood samples. qPCR, western blot analysis, and luciferase assay were performed to confirm the relationship among ERVWE1, cytoplasmic polyadenylation element-binding protein 1 (CPEB1), NADH dehydrogenase ubiquinone flavoprotein 2 (NDUFV2), and NDUFV2 pseudogene (NDUFV2P1). The complex I enzyme activity microplate assay was carried out to evaluate the complex I activity induced by ERVWE1. RESULTS: Herein, we reported decreasing levels of CPEB1 and NDUFV2 in schizophrenia patients. Further studies showed that ERVWE1 was negatively correlated with CPEB1 and NDUFV2 in schizophrenia. Moreover, NDUFV2P1 was increased and demonstrated a significant positive correlation with ERVWE1 and a negative correlation with NDUFV2 in schizophrenia. In vitro experiments disclosed that ERVWE1 suppressed NDUFV2 expression and promoter activity by increasing NDUFV2P1 level. The luciferase assay revealed that ERVWE1 could enhance the promoter activity of NDUFV2P1. Additionally, ERVWE1 downregulated the expression of CPEB1 by suppressing the promoter activity, and the 400 base pair sequence at the 3' terminus of the promoter was the minimum sequence required. Advanced studies showed that CPEB1 participated in regulating the NDUFV2P1/NDUFV2 axis mediated by ERVWE1. Finally, we found that ERVWE1 inhibited complex I activity in SH-SY5Y cells via the CPEB1/NDUFV2P1/NDUFV2 signaling pathway. CONCLUSION: In conclusion, CPEB1 and NDUFV2 might be novel potential blood-based biomarkers and pathogenic factors in schizophrenia. Our findings also reveal a novel mechanism of ERVWE1 in the etiology of schizophrenia.

Characterization of VOC emissions from construction machinery and river ships in the Pearl River Delta of China.

Speciated characterization of Volatile Organic Compounds (VOCs), including oxygenated VOCs (OVOCs), from construction machinery and river ships in China is currently lacking. In this regard, we conducted field measurement on speciated VOC (including OVOC) emissions from six construction machinery and five river ships in the Pearl River Delta (PRD) region to identify VOC emission characteristics. We noticed that OVOC emissions from construction machinery and ships accounted for more than 50% of the total VOC emissions, followed by alkenes, aromatics and alkanes. Formaldehyde and acetaldehyde were the most emission species, accounting for 61.8%-83.2% of OVOCs. For construction machinery, the fuel-based emission factors of roller, grader and pile driver were 3.12, 3.12 and 7.36 g/kg, respectively. With the rigorous restraint by the national emission standards, VOC emissions of construction machinery had decreased considerably, especially during stage Ⅲ. Ozone formation potential was also significantly reduced due to the significant decrease in emissions of OVOCs and alkenes with higher reactivity. For river ships, the fuel-based emission factors of cargo ships and speedboat were 1.46 and 0.44 g/kg, respectively. VOC emissions from construction machinery and river ships in Guangdong Province in 2017 were 8851.0 and 4361.0 ton, respectively. This study filled the knowledge gaps of reactive gas emissions from different kinds of non-road mobile sources over the PRD, and more importantly, highlighted the necessity in adding OVOC measurement to give a complete and accurate depiction of reactive gas emissions from non-road mobile sources.

An Impedance Sensing Platform for Monitoring Heterogeneous Connectivity and Diagnostics in Lab-on-a-Chip Systems.

Reliable hardware connectivity is vital in heterogeneous integrated systems. For example, in digital microfluidics lab-on-a-chip systems, there are hundreds of physical connections required between a microelectromechanical fabricated device and the driving system that can be remotely located on a printed circuit board. Unfortunately, the connection reliability cannot be checked or monitored by vision-based detection methods that are commonly used in the semiconductor industry. Therefore, a sensing platform that can be seamlessly integrated into existing digital microfluidics systems and provide real-time monitoring of multiconnectivity is highly desired. Here, we report an impedance sensing platform that can provide fast detection of a single physical connection in timescales of milliseconds. Once connectivity is established, the same setup can be used to determine the droplet location. The sensing system can be scaled up to support multiple channels or applied to other heterogeneously integrated systems that require real-time monitoring and diagnostics of multiconnectivity systems.

Low fouling and ultrasensitive electrochemical immunosensors with dual assay methods based on Fe3O4 magnetic nanoparticles.

Low fouling electrochemical immunosensors with both "signal-off" and "signal-on" analytical methods were developed for the highly sensitive and efficient detection of cancer antigen 15-3 (CA 15-3) in human serum samples. The antifouling sensing interfaces were constructed by assembling multifunctional polyethylene glycol on gold electrodes, followed by covalent conjugation with CA 15-3 antibody. Pure antigens and Fe3O4@Ag will competitively bind to the immobilized antibody on the electrode. Fe3O4 magnetic nanoparticles attached to the working electrode and collected by a magnetic electrode were treated via electrochemical conversion to generate electroactive Prussian blue as a signal readout. Therefore, these two signals measured independently were complementary, and this design allowed one to choose the assay method according to real situations so as to ensure accuracy of the immunosensor. Moreover, owing to its good antifouling property, the immunosensor was capable of detecting CA 15-3 even in complex human serum samples, demonstrating potential application in quantitative analysis of real patient serum samples.

Low-fouling and highly sensitive fluorescence immunoassay of protein in serum based on the antifouling magnetic beads.

Aim: A low-fouling and highly sensitive fluorescence immunoassay for protein detection in serum was proposed, and IgG was used as a model protein. Materials & methods: SH-PEG-NH2 serving as antifouling coating was conjugated with carboxyl Fe3O4 nanoparticles, and then, the thiol groups were conjugated with antibody via the covalent binding. IgG was captured through magnetic immunoreaction. Highly fluorescent quantum dots modified with streptavidin (SA-QDs) were united with biotin modified IgG antibody to form the sandwich structure. Results & conclusion: The fluorescence immunoassay was able to detect IgG with a detection limit of 3.89 ng/ml in buffer and 5.0 ng/ml in serum with satisfying selectivity and acceptable reproducibility, which demonstrated its potential application in quantitative analysis of real patient serum samples.

High-activity Fe3O4 nanozyme as signal amplifier: A simple, low-cost but efficient strategy for ultrasensitive photoelectrochemical immunoassay.

Sensitive but with simple, inexpensive detection of disease-related biomarkers in real biological samples is of quite necessity for early diagnosis and disease surveillance. We herein first introduced high-activity Fe3O4 nanozyme as signal amplifier to develop an ultrasensitive photoelectrochemical (PEC) immunoassay, which meanwhile has the distinct merits of both simplicity and low cost compared with previously reported enzyme-labeling PEC immunoassays. In the proposal, to illustrate and describe the PEC platform, prostate-specific antigen (PSA, Ag) was used as a target model. Specifically, ZnO nanorods (ZnO-NRs) grown vertically on a bare indium-tin oxide (ITO) electrode was deposited with ZnIn2S4 nanocrystals, producing ZnIn2S4/ZnO-NRs/ITO photoelectrode as the PEC matrix to modify capture PSA antibody (Ab1). Histidine-modified Fe3O4 (his-Fe3O4) nanozyme as signal amplifier was linked with signal PSA antibody (Ab2) to form his-Fe3O4@Ab2 conjugate, and was anchored through specific sandwich immunoreaction. The labeling his-Fe3O4 nanozyme acted as a peroxidase to induce the generation of the insoluble and insulating precipitation, resulting in an evident decrease in the photocurrent signal. On account of combined effects of high catalytic efficiency of the his-Fe3O4 nanozyme and excellent PEC properties of the ZnIn2S4/ZnO-NRs/ITO photoelectrode, ultralow detection limit of 18 fg/mL for target Ag detection was achieved. Besides, as high-activity his-Fe3O4 nanozyme has substituted natural enzyme as signal amplifier, simplicity and low cost of the PEC immunoassay was realized.

Qingkailing Injection () for Treatment of Children Pneumonia Induced by Respiratory Syncytial Virus: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVE: To evaluate the efficacy and safety of Qingkailing Injection (, QKL) for treatment of children pneumonia caused by respiratory syncytial virus (RSV). METHODS: Randomized clinical trials (RCTs) comparing QKL with ribavirin injection in the treatment of children pneumonia induced by RSV were searched in PubMed, Science Direct, Cochrane Library, Chinese VIP database, CNKI and Wanfang databases from their inception to March 2014. Meta-analyses were performed using RevMan 5.2 software. The methodological quality of the selected RCTs was evaluated by the Modified Jadad Score. The primary outcome measures were effective rate and the secondary outcomes were relief time of fever and cough. RESULTS: Seven RCTs with 992 cases published from 2008 to 2013 were identified. The meta-analysis results indicated that QKL was more effective in cure rate [risk ratios (RR)=1.32, 95% CI (1.17, 1.50), P<0.01], total effective rate [RR=1.07, 95% CI (1.02, 1.13), P=0.009] and less fever clearance time [mean difference=-0.73, 95% CI (-1.22,-0.23), P=0.004], compared with ribavirin injection in the treatment of RSV-induced children pneumonia. No dead case was reported in all trials. There were 3 trials mentioned adverse events, 2 reported no obvious adverse event occurred while 1 reported adverse events described as skin hypersensitivity, elevation of ALT, a mild abnormal of hepatic and renal function in both QKL and ribavirin group. CONCLUSIONS: QKL was an effective and relatively safe option for the treatment of RSV-induced children pneumonia. These therapeutic effects were promising but need to be interpreted with caution due to variations in the treatment and methodological weakness in the studies.

Characterize a typically Dscam with alternative splicing in mud crab Scylla paramamosain.

As a member of the immunoglobulin superfamily, Down syndrome cell adhesion molecule (Dscam) could function in the innate immunity of invertebrates. Recently, it is shown that arthropod Dscams play similar functions as antibodies in the adaptive immune system. Dscam could produce thousands of isoforms by alternative splicing and specifically bind to various pathogens. In the present study, we cloned the first Dscam from mud crab Scylla paramamosain (SpDscam), with full-length cDNA 7363 bp containing an open reading frame (ORF) of 6069bp and encoding 2022 amino acids, which had typical domain architecture as other arthropods, i.e., 10 immunoglobulin domains (Ig), 6 fibronectin type 3 domains (FN III), transmembrane and cytoplasmic tail. Quantitative real-time PCR revealed that SpDscam was highly expressed in brain, skin, muscle, intestine and hepatopancreas, but weakly expressed in hemolymph, heart and gill. SpDscam had three alternative splicing regions, located at the N-terminal of Ig2 and Ig3 as well as on the whole Ig7. In these regions, 32, 41 and 14 exons were detected, together with the two exon types of transmembrane domain, indicating SpDscam could potentially encode at least 36,736 unique isoforms. SpDscam induced by Vibrio parahaemolyticus challenge had strong binding ability to V. parahaemolyticus. Further, SpDscam induced by V. parahaemolyticus possessed a clearance of V. parahaemolyticus in S. paramamosain. Collectively, the results indicated SpDscam was a hypervariable pattern-recognition receptor (PRR) by alternative splicing in innate immunity system of mud crab S. paramamosain.

Anisotropic relaxation of a CuO/TiO2 surface under an electric field and its impact on visible light absorption: ab initio calculations.

Ab initio calculations on the anisotropic relaxation of a CuO/TiO2 surface under electric fields and the visible light absorption of these relaxed surfaces are reported. We compare the relaxation of the CuO/TiO2 surface under the electric fields in the direction of [001] or [010]. Fewer Cu-O bonds with highly coordinated Cu-ions are found in the CuO/TiO2 relaxed surface under the electric field in the [010] direction. The Cu-O bonds in the interface of the CuO/TiO2 surface led to an improved visible light absorption in the polarization direction of [001]. The CuO/TiO2 relaxed surface under the electric field in the [010] direction exhibits a more effective absorption of visible light. However, the electric field in the [001] direction induces more relaxation on the CuO/TiO2 surface, breaking the Cu-O bonds. This leads to the partial reduction of CuO to Cu2O on the CuO/TiO2 relaxed surface under the electric field in the [001] direction and inefficient absorption of visible light is observed for this surface.

Combination therapy with VEGFR2 and EGFR siRNA enhances the antitumor effect of cisplatin in non-small cell lung cancer xenografts.

RNA targeting the murine vascular endothelial growth factor receptor 2 (VEGFR2) gene was designed and validated for efficient and robust silencing in vitro and was delivered by polyethylenimines (PEI) in vivo to investigate the antitumor effect on non-small cell lung cancer (NSCLC) xenografts. The following dosage regimens were tested for their tumor inhibitory effect in vivo: VEGFR2 siRNA, epidermal growth factor receptor (EGFR) siRNA, VEGFR2 siRNA+EGFR siRNA, cisplatin alone and VEGFR2 siRNA+ EGFR siRNA+cisplatin. Targeted silencing of both VEGFR2 and EGFR expression by siRNA, combined with low-dose cisplatin, was found to effectively inhibit tumor growth and extend the survival time of mice bearing the NSCLC xenografts. These results suggest that combination therapy using siRNAs and chemotherapy agents might offer a novel strategy for cancer treatment in the future.