Integrating single-nucleus RNA sequencing and spatial transcriptomics to elucidate a specialized subpopulation of astrocytes, microglia and vascular cells in brains of mouse model of lipopolysaccharide-induced sepsis-associated encephalopathy.

BACKGROUND: Understanding the mechanism behind sepsis-associated encephalopathy (SAE) remains a formidable task. This study endeavors to shed light on the complex cellular and molecular alterations that occur in the brains of a mouse model with SAE, ultimately unraveling the underlying mechanisms of this condition. METHODS: We established a murine model using intraperitoneal injection of lipopolysaccharide (LPS) in wild type and Anxa1-/- mice and collected brain tissues for analysis at 0-hour, 12-hour, 24-hour, and 72-hour post-injection. Utilizing advanced techniques such as single-nucleus RNA sequencing (snRNA-seq) and Stereo-seq, we conducted a comprehensive characterization of the cellular responses and molecular patterns within the brain. RESULTS: Our study uncovered notable temporal differences in the response to LPS challenge between Anxa1-/- (annexin A1 knockout) and wild type mice, specifically at the 12-hour and 24-hour time points following injection. We observed a significant increase in the proportion of Astro-2 and Micro-2 cells in these mice. These cells exhibited a colocalization pattern with the vascular subtype Vas-1, forming a distinct region known as V1A2M2, where Astro-2 and Micro-2 cells surrounded Vas-1. Moreover, through further analysis, we discovered significant upregulation of ligands and receptors such as Timp1-Cd63, Timp1-Itgb1, Timp1-Lrp1, as well as Ccl2-Ackr1 and Cxcl2-Ackr1 within this region. In addition, we observed a notable increase in the expression of Cd14-Itgb1, Cd14-Tlr2, and Cd14-C3ar1 in regions enriched with Micro-2 cells. Additionally, Cxcl10-Sdc4 showed broad upregulation in brain regions containing both Micro-2 and Astro-2 cells. Notably, upon LPS challenge, there was an observed increase in Anxa1 expression in the mouse brain. Furthermore, our study revealed a noteworthy increase in mortality rates following Anxa1 knockdown. However, we did not observe substantial differences in the types, numbers, or distribution of other brain cells between Anxa1-/- and wildtype mice over time. Nevertheless, when comparing the 24-hour post LPS injection time point, we observed a significant decrease in the proportion and distribution of Micro-2 and Astro-2 cells in the vicinity of blood vessels in Anxa1-/- mice. Additionally, we noted reduced expression levels of several ligand-receptor pairs including Cd14-Tlr2, Cd14-C3ar1, Cd14-Itgb1, Cxcl10-Sdc4, Ccl2-Ackr1, and Cxcl2-Ackr1. CONCLUSIONS: By combining snRNA-seq and Stereo-seq techniques, our study successfully identified a distinctive cellular colocalization, referred to as a special pathological niche, comprising Astro-2, Micro-2, and Vas-1 cells. Furthermore, we observed an upregulation of ligand-receptor pairs within this niche. These findings suggest a potential association between this cellular arrangement and the underlying mechanisms contributing to SAE or the increased mortality observed in Anxa1 knockdown mice.

Socioeconomic inequalities in human papillomavirus knowledge and vaccine uptake: evidence from a cross-sectional study in China.

Objective: Providing the human papillomavirus (HPV) vaccine is effective to eliminate the disparity in HPV-related cancers. It is unknown regarding inequality in the distribution of HPV vaccination in China since the vaccine was licensed and approved for use in 2016. This study aimed to examine socioeconomic inequalities in HPV-related knowledge and vaccination and identified factors associated with such inequalities. Methods: Self-administered questionnaires measuring HPV-related knowledge and vaccine uptake were completed by 1,306 women through online survey platform. HPV knowledge was assessed using a 12-item question stem that covered the hazards of HPV infection, HPV vaccine dosage, benefits, and protection. Cluster analysis by combining monthly household income, educational level, and employment status was used to identify socioeconomic status (SES) class. The concentration index (CI) was employed as a measure of socioeconomic inequalities in HPV-related knowledge and vaccination. Linear regression and logistic regression were established to decompose the contributions of associated factors to the observed inequalities. Results: The CI for HPV-related knowledge and vaccine uptake was 0.0442 and 0.1485, respectively, indicating the higher knowledge and vaccination rate were concentrated in groups with high SES. Education and household income made the largest contribution to these inequalities. Age, residency and cervical cancer screening were also important contributors of observed inequalities. Conclusion: Socioeconomic inequalities in HPV-related knowledge and vaccination uptake are evident in China. Interventions to diffuse HPV-related information for disadvantaged groups are helpful to reduce these inequalities. Providing low or no-cost HPV vaccination and ensuring accessibility of vaccines in rural areas are also considered to be beneficial.

Compared with soil fungal diversity and microbial network complexity, soil bacterial diversity drives soil multifunctionality during the restoration process.

Understanding factors driving soil multifunctionality can help with terrestrial ecosystem restoration. Soil microbial diversity and network complexity are two important factors influencing ecosystem multifunctionality. However, their effects on soil multifunctionality are still unclear. Based on high-throughput sequencing, we analyzed soil microbial alpha diversity and network complexity and their relative impacts on soil multifunctionality during the aerial seeding restoration process from 1983 to 2017 in Mu Us sandy land, China, a region threatened by desertification. Our results showed soil bacterial and fungal alpha diversity and multifunctionality increased with aerial seeding restoration. We found the community composition of soil bacteria and fungi changed with restoration periods. The keystone species of the soil bacterial network changed during restoration, while those of the soil fungal network remained unchanged. Soil bacterial and fungal species mainly maintained positive associations throughout the restoration periods. Soil bacterial network complexity initially decreased before increasing with restoration, while soil fungal network complexity increased continuously. Soil multifunctionality was found to have significantly positive correlations with soil fungal network complexity and soil bacterial alpha diversity. Compared with soil fungal alpha diversity and soil microbial network complexity, soil bacterial alpha diversity significantly promoted soil multifunctionality. Our research highlights the critical impact that soil bacterial alpha diversity plays in soil multifunctionality in restored ecosystems threatened by desertification.

Carpaine alleviates tendinopathy in mice by promoting the ubiquitin-proteasomal degradation of p65 via targeting the E3 ubiquitin ligase LRSAM1.

BACKGROUND: Currently, there are no specific drugs or targets available for the treatment of tendinopathy. However, inflammation has recently been found to play a pivotal role in tendinopathy progression, thereby identifying it as a potential therapeutic target. Carpaine (CA) exhibits potential anti-inflammatory pharmacological properties and may offer a therapeutic option for tendinopathy. PURPOSE: This study aimed to investigate the effectiveness of CA in addressing tendinopathy and uncovering its underlying mechanisms. METHODS: Herein, the efficacy of CA by local administration in vivo in comparison to the first-line drug indomethacin was evaluated in a mouse collagenase-induced tendinopathy (CIT) model. Furthermore, IL-1ß induced a simulated pathological inflammatory microenvironment in tenocytes to investigate its underlying mechanisms in vitro. Further confirmation experiments were performed by overexpressing or knocking down the selective targets of CA in vivo. RESULTS: The findings demonstrated that CA was dose-dependent in treating tendinopathy and that the high-dose group outperformed the first-line drug indomethacin. Mechanistically, CA selectively bound to and enhanced the activity of the E3 ubiquitin ligase LRSAM1 in tendinopathy. This effect mediated the ubiquitination of p65 at lysine 93, subsequently promoting its proteasomal degradation. As a result, the NF-κB pathway was inactivated, leading to a reduction in inflammation of tendinopathy. Consequently, CA effectively mitigated the progression of tendinopathy. Moreover, the LRSAM1 overexpression demonstrated effectiveness in mitigating the tendinopathy progression and its knockdown abolished the therapeutic effects of CA. CONCLUSION: CA attenuates the progression of tendinopathy by promoting the ubiquitin-proteasomal degradation of p65 via increasing the enzyme activity of LRSAM1. The exploration of LRSAM1 has also unveiled a new potential target for treating tendinopathy based on the ubiquitin-proteasomal pathway.

Pristimerin suppresses AIM2 inflammasome by modulating AIM2-PYCARD/ASC stability via selective autophagy to alleviate tendinopathy.

Macroautophagy/autophagy plays an important role in regulating cellular homeostasis and influences the pathogenesis of degenerative diseases. Tendinopathy is characterized by tendon degeneration and inflammation. However, little is known about the role of selective autophagy in tendinopathy. Here, we find that pristimerin (PM), a quinone methide triterpenoid, is more effective in treating tendinopathy than the first-line drug indomethacin. PM inhibits the AIM2 inflammasome and alleviates inflammation during tendinopathy by promoting the autophagic degradation of AIM2 through a PYCARD/ASC-dependent manner. A mechanistic study shows that PM enhances the K63-linked ubiquitin chains of PYCARD/ASC at K158/161, which serves as a recognition signal for SQSTM1/p62-mediated autophagic degradation of the AIM2-PYCARD/ASC complex. We further identify that PM binds the Cys53 site of deubiquitinase USP50 through the Michael-acceptor and blocks the binding of USP50 to PYCARD/ASC, thereby reducing USP50-mediated cleavage of K63-linked ubiquitin chains of PYCARD/ASC. Finally, PM treatment in vivo generates an effect comparable to inflammasome deficiency in alleviating tendinopathy. Taken together, these findings demonstrate that PM alleviates the progression of tendinopathy by modulating AIM2-PYCARD/ASC stability via SQSTM1/p62-mediated selective autophagic degradation, thus providing a promising autophagy-based therapeutic for tendinopathy.Abbreviations: 3-MA: 3-methyladenine; AIM2: absent in melanoma 2; AT: Achilles tenotomy; ATP: adenosine triphosphate; BMDMs: bone marrow-derived macrophages; CHX: cycloheximide; Col3a1: collagen, type III, alpha 1; CQ: chloroquine; Cys: cysteine; DARTS: drug affinity responsive target stability; DTT: dithiothreitol; DUB: deubiquitinase; gDNA: genomic DNA; GSH: glutathione; His: histidine; IL1B/IL-1ß: interleukin 1 beta; IND: indomethacin; IP: immunoprecipitation; LPS: lipopolysaccharide; MMP: mitochondrial membrane potential; NLRP3: NLR family, pyrin domain containing 3; PM: pristimerin; PYCARD/ASC: PYD and CARD domain containing; SN: supernatants; SOX9: SRY (sex determining region Y)-box 9; SQSTM1: sequestosome 1; Tgfb: transforming growth factor, beta; TIMP3: tissue inhibitor of metalloproteinase 3; TNMD: tenomodulin; TRAF6: TNF receptor-associated factor 6; Ub: ubiquitin; USP50: ubiquitin specific peptidase 50; WCL: whole cell lysates.

Promoting effects of soil C and N and limiting effect of soil P jointly determine the plant diversity during the aerial seeding restoration process in Mu Us sandy land, China.

Introduction: Exploring the change and maintaining mechanism of plant diversity is of great significance for guiding the restoration of degraded ecosystems. However, how plant taxonomic, functional, and phylogenetic diversity change during long-term ecosystem restoration process and their driving factors remain unclear. Methods: Based on the 35-year time gradient of aerial seeding restoration in Mu Us sandy land, this study explored the changes in plant taxonomic, functional, and phylogenetic diversity and the driving factors. Results: The results showed that plant taxonomic, functional, and phylogenetic diversity showed consistent response with the aerial seeding restoration, all of which increased first and then tended to a saturation state in the middle of restoration (14 years). TN, TOC, and NO3 --N increased with aerial seeding restoration and showed a significant positive correlation with plant diversity of the three dimensions, while AP showed a negative correlation. Soil nitrogen and carbon promoted the increase of diversity of three dimensions in the early restoration period, while phosphorus limited the increase of diversity of three dimensions in the middle and late restoration periods. The diversity of three dimensions was mainly affected by restoration time, soil nutrients, and climate factors, and the coupling effect of restoration time and soil nutrients was dominant. Discussion: These findings indicate that the plant diversity in different dimensions and soil nutrients are improved by aerial seeding restoration. Our study highlights that aerial seeding restoration mainly improves plant diversity by increasing soil nutrients, and the relative effects of different soil nutrients on plant diversity during restoration are inconsistent.

Erythropoietin relieves neuronal apoptosis in epilepsy rats via TGF-ß/Smad signaling pathway.

This study aimed to investigate the influence of recombinant human erythropoietin (rHuEPO) on pentylenetetrazol (PTZ)-induced neuronal apoptosis in epilepsy rats, and to explore the signaling pathways related to the action. Healthy Sprague-Dawley rats aged 8 weeks old were randomly divided into 5 groups, namely, control group, PTZ model group, PTZ + rHuEPO intervention group, PTZ + SB431542 + rHuEPO intervention group and PTZ + SB431542 (TGF-ß/Smad inhibitor) intervention group. The expressions of apoptotic proteins [tumor necrosis factor receptor 1 (TNFR1) and caspase-3] and the transforming growth factor-beta (TGF-ß)/Smad signaling pathway-related proteins [phosphorylated smad3 (p-smad3) and TGF-ß1] in the brain tissues were determined via Western blotting (WB). Epilepsy was successfully induced by PTZ in the rats. The results of the TUNEL assay showed that the intervention with rHuEPO could remarkably reduce the number of PTZ-induced apoptotic neurons in the hippocampus, while SB431542 inhibitor could attenuate the protective effect of rHuEPO against neuronal apoptosis (P<0.05). In addition, the intraperitoneal injection of 50 µg/kg rHuEPO could activate the TGF-ß/Smad signaling pathway, markedly up-regulate the expressions of TGF-ß1 and p-smad3 (P<0.05), down-regulate the expressions of apoptotic proteins TNFR1 and caspase-3 (P<0.01) and reduce neuronal apoptosis. Moreover, SB431542 was able to notably repress the protective effect of rHuEPO against neuronal apoptosis, and down-regulate the expressions of p-smad3 and TGF-ß1 (P<0.01). In conclusion, the inhibitory effect of rHuEPO on nerve cell apoptosis in epilepsy rats may be realized by activating the TGF-ß/Smad signaling pathway, thus relieving neuronal apoptosis and ameliorating the symptoms of epilepsy.

Real-world clinical outcomes of oral anticoagulants among Japanese patients with atrial fibrillation and concomitant coronary artery disease.

Background: Stroke prevention is complicated in patients with atrial fibrillation (AF) and coronary artery disease (CAD). We compared the risk of major bleeding among Japanese patients with AF and CAD commencing warfarin, dabigatran, or rivaroxaban. Methods: This study included adults with AF and CAD who were newly prescribed the non-vitamin K antagonist oral anticoagulants (NOACs) dabigatran or rivaroxaban, or warfarin, and registered between 18 April 2011 through 31 December 2020 in the Medical Data Vision hospital-based clinical database. The primary outcome was major bleeding, and the secondary outcome was a composite of stroke, systemic embolism, myocardial infarction, all-cause inpatient mortality, major bleeding, major gastrointestinal bleeding, and intracerebral hemorrhage. Cox proportional hazard models with stabilized inverse probability treatment weighting were used to estimate hazard ratios (HRs) with 95 % CIs via a two-step approach; first between warfarin and each NOAC, then between NOACs if sample size conditions were met. Results: Dabigatran, rivaroxaban, and warfarin groups included 6712, 20,329, and 12,316 patients, respectively. Major bleeding risk was lower in NOACs versus warfarin (dabigatran: HR 0.50, 95 % CI: 0.40-0.62; rivaroxaban: HR 0.78, 95 % CI: 0.69-0.90); this risk was lower with dabigatran compared with rivaroxaban (HR 0.64, 95 % CI: 0.51─0.79). Net clinical benefit was superior to warfarin in both NOACs (dabigatran: HR 0.78, 95 % CI: 0.71-0.85; rivaroxaban: HR 0.83, 95 % CI: 0.78-0.88). Conclusions: Among real-world Japanese patients with AF and CAD, NOACs were associated with better clinical outcomes than warfarin. Treatment with dabigatran had a lower risk of major bleeding than rivaroxaban.Clinical trial registration: NCT05051904 (ClinicalTrials.gov).

Attitudes and associated factors of patients' adoption of patient accessible electronic health records in China - A mixed methods study.

Background: Although patient accessible electronic health records (PAEHRs) offer great potential in enhancing the provision of patient-centered care and improving satisfaction, the adoption rate is still low. Currently, few studies are there for researchers and health organization leaders to understand patients' thoughts and related factors of PAEHRs adoption in developing countries. China adopted more limited practices of PAEHRs, among which we selected Yuebei People's Hospital as an example. Objective: The study aimed to research patient attitudes toward PAEHRs use and the associated factors of patients' adoption of PAEHRs in China, which is achieved by both qualitative and quantitative studies. Methods: This study employed sequential mixed-methods. The DeLone & McLean information systems (D&M IS) success model, Unified Theory of Acceptance and Use of Technology (UTAUT) and task-technology fit (TTF) model were used to guide the research. Finally, we collected 28 valid in-depth interview responses, 51 valid semi-structured interview responses and 235 valid questionnaire responses. The research model was tested and validated using data collected. Results: The findings of the qualitative study reveal that patients' rate perceived task productivity and customer satisfaction as benefits, and poor-quality information as flaws. Results of the quantitative study show that the drivers of behavioral intention are performance expectance, effort expectancy and social influence; the predictors of use behavior are TTF and behavioral intention. Conclusion: It is necessary to consider PAEHRs' task-tool role in patients' adoption behavior. Hospitalized patients value PAEHRs' practical attributes and attach much importance to the information content and application design.

Modeling the role of land conversion on the spread of an epizootic disease.

Land conversion and the resulting contact between domesticated and wild species has arguably been the single largest contributor to the emergence of novel epizootic and zoonotic diseases in the past century. An unintended consequence of these interactions is zoonotic or epizootic disease spillovers from wild species to humans and their domesticates. Disease spillovers are edge effects of land conversion and are sensitive to the size and shape of converted areas. We combine spatial metrics from landscape ecology with theoretical epidemiological models to understand how the size and shape of land conversion affect epizootic and zoonotic disease transmission of single and two species populations. We show that the less compact the converted area, and the greater the depth of the contact zone, the more rapidly will an introduced disease spread through the domesticated population.

Accuracy of Vegetation Indices in Assessing Different Grades of Grassland Desertification from UAV.

Grassland desertification has become one of the most serious environmental problems in the world. Grasslands are the focus of desertification research because of their ecological vulnerability. Their application on different grassland desertification grades remains limited. Therefore, in this study, 19 vegetation indices were calculated for 30 unmanned aerial vehicle (UAV) visible light images at five grades of grassland desertification in the Mu Us Sandy. Fractional Vegetation Coverage (FVC) with high accuracy was obtained through Support Vector Machine (SVM) classification, and the results were used as the reference values. Based on the FVC, the grassland desertification grades were divided into five grades: severe (FVC < 5%), high (FVC: 5-20%), moderate (FVC: 21-50%), slight (FVC: 51-70%), and non-desertification (FVC: 71-100%). The accuracy of the vegetation indices was assessed by the overall accuracy (OA), the kappa coefficient (k), and the relative error (RE). Our result showed that the accuracy of SVM-supervised classification was high in assessing each grassland desertification grade. Excess Green Red Blue Difference Index (EGRBDI), Visible Band Modified Soil Adjusted Vegetation Index (V-MSAVI), Green Leaf Index (GLI), Color Index of Vegetation Vegetative (CIVE), Red Green Blue Vegetation Index (RGBVI), and Excess Green (EXG) accurately assessed grassland desertification at severe, high, moderate, and slight grades. In addition, the Red Green Ratio Index (RGRI) and Combined 2 (COM2) were accurate in assessing severe desertification. The assessment of the 19 indices of the non-desertification grade had low accuracy. Moreover, our result showed that the accuracy of SVM-supervised classification was high in assessing each grassland desertification grade. This study emphasizes that the applicability of the vegetation indices varies with the degree of grassland desertification and hopes to provide scientific guidance for a more accurate grassland desertification assessment.

Peptidomics Analysis Reveals Serum Biomarkers in Spinal Cord Injury Patients.

Spinal cord injury (SCI) is a severe traumatic disease, always resulting in neuronal injury. In this study, we aimed to exhibit a peptidome profile of serum from patients with SCI. A label-free peptidomics strategy was used to analyze the differentially expressed peptides (DEPs). Then, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses was used to evaluate the function of the peptides precursors proteins. Also, the protein-protein interaction networks were mapped using STRING database. Finally, parallel reaction monitoring assays were used to validate the expression of candidate peptides. We identified 217 DEPs including 29 upregulated peptides and 188 downregulated peptides in SCI group. Many pathways such as Platelet activation, Complement and coagulation cascades, Focal adhesion were enriched. Seven peptides including PSPRPSP, RPPGFSP, DKPDMAEIEKFDKSKLK, STTAVVTNPKE, GHAGAQGPPGPPG, SMPPAQQQITS and SKVLPIQDNVSK were significantly changed between SCI patients and healthy people. Peptidomics provide a powerful tool to find the variation of SCI. RPPGFSP, DKPDMAEIEKFDKSKLK and SMPPAQQQITS may play important roles in SCI. However, the specific function of these peptides and whether they can be used as therapeutic targets for SCI need to be further investigated.

The Effectiveness of Repetitive Transcranial Magnetic Stimulation for Post-stroke Dysphagia: A Systematic Review and Meta-Analysis.

Background: Repetitive transcranial magnetic stimulation (rTMS) applied to the mylohyoid cortical region has positive clinical effects on post-stroke. Therefore, we conducted a meta-analysis to investigate the efficacy of rTMS for patients with post-stroke dysphagia. Methods: According to PRISMA guidelines, we searched the databases of MEDLINE (PubMed), Cochrane Library, Embase, Web of Science, CNKI, Wangfang. We searched for studies of randomized controlled trials (RCTs) of rTMS to treat dysphagia after stroke and screened by inclusion and exclusion criteria. Features of RCTs were extracted. The heterogeneity of the trials was measured by I 2 statistic. Results: In total, 11 RCTs with 463 dysphagia patients fulfilled our inclusion criteria. In our analysis, rTMS demonstrated a great beneficial effect for post-stroke dysphagia when combined with traditional swallowing exercises. Moreover, a greatly significant difference (P = 0.008) was noted based on stimulation frequency (high frequency vs. low frequency). Additionally, no significant difference (P = 0.53) was observed based on stimulation site (affected vs. unaffected hemisphere). Conclusions: Overall, rTMS can effectively accelerate the improvement of swallowing function in patients with post-stroke swallowing disorders.

The Application of Brain-Computer Interface in Upper Limb Dysfunction After Stroke: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Objective: This study aimed to examine the effectiveness and safety of the Brain-computer interface (BCI) in treatment of upper limb dysfunction after stroke. Methods: English and Chinese electronic databases were searched up to July 2021. Randomized controlled trials (RCTs) were eligible. The methodological quality was assessed using Cochrane's risk-of-bias tool. Meta-analysis was performed using RevMan 5.4. Results: A total of 488 patients from 16 RCTs were included. The results showed that (1) the meta-analysis of BCI-combined treatment on the improvement of the upper limb function showed statistical significance [standardized mean difference (SMD): 0.53, 95% CI: 0.26-0.80, P < 0.05]; (2) BCI treatment can improve the abilities of daily living of patients after stroke, and the analysis results are statistically significant (SMD: 1.67, 95% CI: 0.61-2.74, P < 0.05); and (3) the BCI-combined therapy was not statistically significant for the analysis of the Modified Ashworth Scale (MAS) (SMD: -0.10, 95% CI: -0.50 to 0.30, P = 0.61). Conclusion: The meta-analysis indicates that the BCI therapy or BCI combined with other therapies such as conventional rehabilitation training and motor imagery training can improve upper limb dysfunction after stroke and enhance the quality of daily life.

Transcranial Direct Current Stimulation for Parkinson's Disease: A Systematic Review and Meta-Analysis.

BACKGROUND: Parkinson's disease is a common neurodegenerative disorder with motor and non-motor symptoms. Recently, as adjuvant therapy, transcranial direct current stimulation (tDCS) has been shown to improve the motor and non-motor function of patients with Parkinson's disease (PD). This systematic review aimed to evaluate the existing evidence for the efficacy of tDCS for PD. We included English databases (PubMed, the Cochrane Library, Embase, and Web of Science) and Chinese databases [Wanfang database, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), and China Biology Medicine (CBM)] without restricting the year of publication. Twenty-one tDCS studies, with a total of 736 participants, were included in the analysis. Two independent researchers extracted the data and characteristics of each study. There was a significant pooled effect size (-1.29; 95% CI = -1.60, -0.98; p < 0.00001; I 2 = 0%) in the Unified PD Rating Scale (UPDRS) I and the Montreal cognitive assessment (SMD = 0.87, 95% CI = 0.50 to 1.24; p < 0.00001; I 2 = 0%). The poor effect size was observed in the UPDRS III scores (SMD = -0.13; 95% CI = -0.64, 0.38; p = 0.61; I 2 = 77%), and similar results were observed for the timed up and go (TUG) test, Berg balance scale, and gait assessment. The results of this meta-analysis showed that there was insufficient evidence that tDCS improves the motor function of patients with PD. However, tDCS seemed to improve their cognitive performance. Further multicenter research with a larger sample size is needed. In addition, future research should focus on determining the tDCS parameters that are most beneficial to the functional recovery of patients with PD.

Inhibition of anti-viral stress granule formation by coronavirus endoribonuclease nsp15 ensures efficient virus replication.

Cytoplasmic stress granules (SGs) are generally triggered by stress-induced translation arrest for storing mRNAs. Recently, it has been shown that SGs exert anti-viral functions due to their involvement in protein synthesis shut off and recruitment of innate immune signaling intermediates. The largest RNA viruses, coronaviruses, impose great threat to public safety and animal health; however, the significance of SGs in coronavirus infection is largely unknown. Infectious Bronchitis Virus (IBV) is the first identified coronavirus in 1930s and has been prevalent in poultry farm for many years. In this study, we provided evidence that IBV overcomes the host antiviral response by inhibiting SGs formation via the virus-encoded endoribonuclease nsp15. By immunofluorescence analysis, we observed that IBV infection not only did not trigger SGs formation in approximately 80% of the infected cells, but also impaired the formation of SGs triggered by heat shock, sodium arsenite, or NaCl stimuli. We further demonstrated that the intrinsic endoribonuclease activity of nsp15 was responsible for the interference of SGs formation. In fact, nsp15-defective recombinant IBV (rIBV-nsp15-H238A) greatly induced the formation of SGs, along with accumulation of dsRNA and activation of PKR, whereas wild type IBV failed to do so. Consequently, infection with rIBV-nsp15-H238A strongly triggered transcription of IFN-ß which in turn greatly affected rIBV-nsp15-H238A replication. Further analysis showed that SGs function as an antiviral hub, as demonstrated by the attenuated IRF3-IFN response and increased production of IBV in SG-defective cells. Additional evidence includes the aggregation of pattern recognition receptors (PRRs) and signaling intermediates to the IBV-induced SGs. Collectively, our data demonstrate that the endoribonuclease nsp15 of IBV interferes with the formation of antiviral hub SGs by regulating the accumulation of viral dsRNA and by antagonizing the activation of PKR, eventually ensuring productive virus replication. We further demonstrated that nsp15s from PEDV, TGEV, SARS-CoV, and SARS-CoV-2 harbor the conserved function to interfere with the formation of chemically-induced SGs. Thus, we speculate that coronaviruses employ similar nsp15-mediated mechanisms to antagonize the host anti-viral SGs formation to ensure efficient virus replication.

The role of PA-X C-terminal 20 residues of classical swine influenza virus in its replication and pathogenicity.

PA-X is a fusion protein encoded by a +1 frameshifted open reading frame (X-ORF) in PA gene. The X-ORF can be translated in full-length (61 amino acids, aa) or truncated (41 aa) form. However, the role of C-Terminal 20 aa of PA-X in virus function has not yet been fully elucidated. To this end, we constructed the contemporary influenza viruses with full and truncated PA-X by reverse genetics to compare their replication and pathogenicity. The full-length PA-X virus in MDCK and human A549 cells conferred 10- to 100-fold increase in viral replication, and more virulent and caused more severe inflammatory responses in mice relative to corresponding truncated PA-X virus, suggesting that the terminal 20 aa could play a role in enhancing viral replication and contribute to virulence.

Protective efficacy of a bivalent inactivated reassortant H1N1 influenza virus vaccine against European avian-like and classical swine influenza H1N1 viruses in mice.

The classical swine (CS) H1N1 swine influenza virus (SIVs) emerged in humans as a reassortant virus that caused the H1N1 influenza virus pandemic in 2009, and the European avian-like (EA) H1N1 SIVs has caused several human infections in European and Asian countries. Development of the influenza vaccines that could provide effective protective efficacy against SIVs remains a challenge. In this study, the bivalent reassortant inactivated vaccine comprised of SH1/PR8 and G11/PR8 arboring the hemagglutinin (HA) and neuraminidase (NA) genes from prevalent CS and EA H1N1 SIVs and six internal genes from the A/Puerto Rico/8/34(PR8) virus was developed. The protective efficacy of this bivalent vaccine was evaluated in mice challenged with the lethal doses of CS and EA H1N1 SIVs. The result showed that univalent inactivated vaccine elicited high-level antibody against homologous H1N1 viruses while cross-reactive antibody responses to heterologous H1N1 viruses were not fully effective. In a mouse model, the bivalent inactivated vaccine conferred complete protection against lethal challenge doses of EA SH1 virus or CS G11 virus, whereas the univalent inactivated vaccine only produced insufficient protection against heterologous SIVs. In conclusion, our data demonstrated that the reassortant bivalent inactivated vaccine comprised of SH1/PR8 and G11/PR8 could provide effective protection against the prevalent EA and CS H1N1 subtype SIVs in mice.

Melatonin Attenuates Thrombin-induced Inflammation in BV2 Cells and Then Protects HT22 Cells from Apoptosis.

Increasing evidence has revealed that the uncontrolled thrombin-induced inflammation following intracerebral hemorrhage (ICH) plays a key role in ICH. Oxidative stress and neuroinflammatory responses are interdependent and bidirectional events. Melatonin is now recognized as an antioxidant and a free radical scavenger due to its roles in various physiological and pathological processes. The aim of this study was to explore the molecular mechanisms underlying the effects of melatonin on thrombin-induced microglial inflammation and its indirect protection of HT22 cells from p53-associated apoptosis. Melatonin treatment attenuated the expression of IL-1ß, IL-18, cleaved caspase-1, and NLRP3 and decreased the production of reactive oxygen species (ROS), revealing its inhibitory effects against ROS-NLRP3 inflammasome activation. In further experiments investigating the protection conferred by melatonin, incubating HT22 cells with conditioned medium (CM) from thrombin-stimulated microglia induced HT22 cell apoptosis, and this effect was reversed after treating CM with either melatonin or N-acetyl-L-cysteine (NAC). Additionally, the Bax/Bcl-2 ratio and the levels of cleaved caspase-3 and p53 were markedly lower in the cells cultured in thrombin + melatonin-CM than in the cells cultured in thrombin-CM. Furthermore, the levels of MMP, ROS, SOD, MDA, and GSH-PX in bystander HT22 cells suggested that melatonin decreased HT22 cell apoptosis instigated via the p53-associated apoptotic pathway. Therefore, these findings strongly indicate the anti-inflammatory properties of melatonin that may suppress ROS-NLRP3 inflammasome activation and protect HT22 cells against apoptosis by inhibiting the ROS-mediated p53-dependent mitochondrial apoptotic pathway.

Long non­coding RNA LINC00312 regulates breast cancer progression through the miR­9/CDH1 axis.

Long non­coding RNAs (lncRNAs) are important mediators of the initiation and progression of tumors, including breast cancer (BC). The exact role of long intergenic non­coding RNA 00312 (LINC00312) in BC and its mechanism of action have not been reported to date. In the present study, LINC00312 was found to be downregulated in human BC tissues and cell lines by RT­qPCR. The findings of a functional study indicated that overexpression of LINC00312 suppressed the proliferation, colony forming ability, migration and invasiveness of BC cell lines. Mechanistically, LINC00312 was found to induce suppression of cell migration and invasion by directly binding to miR­9. Overexpression of LINC00312 increased the expression of cadherin 1 (CDH1), a direct target of miR­9, and decreased the expression of vimentin (VIM), a major cytoskeletal component of mesenchymal cells as determined by western blot analysis. miR­9 partly abrogated the upregulation of CDH1 and downregulation of VIM induced by LINC00312. Taken together, the results of the present study indicate a role for the LINC00312/miR­9/CDH1 axis in the progression of BC, and suggest a novel lncRNA­based diagnostic biomarker or therapeutic target for BC.