An extended conformation of SARS-CoV-2 main protease reveals allosteric targets.

SignificanceThe coronavirus main protease (Mpro) is required for viral replication. Here, we obtained the extended conformation of the native monomer of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Mpro by trapping it with nanobodies and found that the catalytic domain and the helix domain dissociate, revealing allosteric targets. Another monomeric state is termed compact conformation and is similar to one protomer of the dimeric form. We designed a Nanoluc Binary Techonology (NanoBiT)-based high-throughput allosteric inhibitor assay based on structural conformational change. Our results provide insight into the maturation, dimerization, and catalysis of the coronavirus Mpro and pave a way to develop an anticoronaviral drug through targeting the maturation process to inhibit the autocleavage of Mpro.

Structure basis of two nanobodies neutralizing SARS-CoV-2 Omicron variant by targeting ultra-conservative epitopes.

The evolving SARS-CoV-2 Omicron strain has repeatedly caused widespread disease epidemics, and effective antibody drugs continue to be in short supply. Here, we identified a batch of nanobodies with high affinity for receptor binding domain (RBD) of SARS-CoV-2 spike protein, separated them into three classes using high performance liquid chromatography (HPLC), and then resolved the crystal structure of the ternary complexes of two non-competing nanobodies (NB1C6 and NB1B5) with RBD using X-ray crystallography. The structures showed that NB1B5 and NB1C6 bind to the left and right flank of the RBD, respectively, and that the binding epitopes are highly conserved cryptic sites in all SARS-CoV-2 mutant strains, as well as that NB1B5 can effectively block the ACE2. These two nanobodies were covalently linked into multivalent and bi-paratopic formats, and have a high affinity and neutralization potency for omicron, potentially inhibiting viral escape. The binding sites of these two nanobodies are relatively conserved, which help guide the structural design of antibodies targeting future variants of SARS-CoV-2 to combat COVID-19 epidemics and pandemics.

GAS6/Axl is associated with AMPK activation and attenuates H2O2-induced oxidative stress.

Oxidative stress plays a key part in cardiovascular event. Growth arrest-specific gene 6 (GAS6) is a vitamin K-dependent ligand which has been shown to exert important effects in heart. The effects of GAS6 were evaluated against hydrogen peroxide (H2O2) ­induced oxidative stress injury in HL-1 cardiomyocytes. A series of experimental methods were used to analyze the effects of GAS6 on cell viability, apoptosis, oxidative stress, mitochondrial function and AMPK/ACC signaling in H2O2­injured HL-1 cells. In this study, we found that H2O2 reduced cell viability, increased apoptotic rate and intracellular reactive oxygen species (ROS). Meanwhile, H2O2 decreased the protein levels of GAS6, and increased the protein level of p-AMPK/AMPK, p-ACC/ACC. Then, we observed that overexpression of GAS6 significantly reduced cell death, manifested as increased cell viability, improved oxidative stress, apoptosis and upregulated the levels of GAS6, p-Axl/Axl, Nrf2, NQO1, HO-1, Bcl-2/Bax, PGC-1α, NRF1, TFAM, p-AMPK/AMPK, and p-ACC/ACC-related protein expression in HL-1 cells and H2O2­injured cardiomyocytes. To further verify the results, we successfully constructed GAS6 lentiviral vectors, and found GAS6 shRNA partially reversed the above results. These data suggest that AMPK/ACC may be a downstream effector molecule in the antioxidant action of GAS6. In summary, our findings indicate that activation GAS6/Axl-AMPK signaling protects H2O2­induced oxidative stress which is accompanied by the amelioration of oxidative stress, apoptosis, and mitochondrial function.

CTRP family in diseases associated with inflammation and metabolism: molecular mechanisms and clinical implication.

C1q/tumor necrosis factor (TNF) related proteins (CTRPs) is a newly discovered adipokine family with conservative structure and ubiquitous distribution and is secreted by adipose tissues. Recently, CTRPs have attracted increasing attention due to the its wide-ranging effects upon inflammation and metabolism. To-date, 15 members of CTRPs (CTRP1-15) with the characteristic C1q domain have been characterized. Earlier in-depth phenotypic analyses of mouse models of CTRPs deficiency have also unveiled ample function of CTRPs in inflammation and metabolism. This review focuses on the rise of CTRPs, with a special emphasis on the latest discoveries with regards to the effects of the CTRP family on inflammation and metabolism as well as related diseases. We first introduced the structure of characteristic domain and polymerization of CTRPs to reveal its pleiotropic biological functions. Next, intimate association of CTRP family with inflammation and metabolism, as well as the involvement of CTRPs as nodes in complex molecular networks, were elaborated. With expanding membership of CTRP family, the information presented here provides new perspectives for therapeutic strategies to improve inflammatory and metabolic abnormalities.

The association of dietary inflammatory potential with sarcopenia in Chinese community-dwelling older adults.

BACKGROUND: Dietary-induced inflammation is potentially associated with sarcopenia. Nevertheless, few studies have investigated the structure of the inflammatory diet and its correlation with muscle function and performance in both the upper and lower limbs. This study was performed to explore the association of the dietary inflammatory index (DII) with sarcopenia and its diagnostic parameters. METHODS: We conducted a cross-sectional survey on a sample of 515 Chinese community-dwelling older adults selected through multistage cluster sampling from three districts in Shanghai. DII scores were calculated using a validated food frequency questionnaire. Sarcopenia and its diagnostic parameters were determined based on the definition set by the Asian Working Group on Sarcopenia (AWGS). RESULTS: The mean age of study participants was 71.31 ± 4.71 years. The prevalence of sarcopenia in the cohort was 12.4%. Older adults in the highest DII quartile had a 3.339 times increased risk of sarcopenia compared to those in the lowest quartile (OR Quartile4vs1:3.339, 95%CI: 1.232, 9.052, p-trend: 0.004) after adjusting for confounding factors. Additionally, a more pro-inflammatory diet was associated with lower appendicular skeletal muscle index (ASMI) (OR Quartile4vs1: 3.005, 95%CI: 1.275, 7.318, p-trend: 0.005), a higher 5-times sit-stand test time score (OR Quartile4vs1: 4.942, 95%CI: 1.745, 13.993, p-trend: 0.005), and lower gait speed (OR Quartile4vs1: 2.392, 95%CI: 1.104, 5.185, p-trend: 0.041) after adjusting for confounding factors. However, there was no significant association between DII, handgrip strength, and Short Physical Performance Battery (SPPB) score in either the unadjusted or adjusted model. CONCLUSION: This study found that the association between consuming a more pro-inflammatory diet and sarcopenia in Chinese community-dwelling older adults was mainly due to underlying low intakes of dietary energy, protein, and anti-inflammatory foods, and not due to the high intake of pro-inflammatory foods. Meanwhile, DII was more highly correlated with lower limb muscle strength and performance compared to upper limb muscle strength.

Development and validity of computerized neuropsychological assessment devices for screening mild cognitive impairment: Ensemble of models with feature space heterogeneity and retrieval practice effect.

OBJECTIVE: This study aimed to develop and validate computerized neuropsychological assessment devices for screening patients with mild cognitive impairment (MCI). METHODS: We conducted this study in three phases. Phase I involved the development of a conceptual framework of Memory Guard (MG) based on the principles of the cognitive design system (CDS). Phase II involved three steps of feature engineering: item development, filter, and wrapper. Based on the initial items, the number of items in each dimension was determined through analytic hierarchy process. We constructed an initial set with a total of 198 items with three levels of difficulty. Next, we performed feature selection through comprehensive reliability and validity tests, which resulted in the best item bank of 38 test items. The features for modeling were obtained from the best item bank (option scores, reading time scores and total time scores), demographic variables and their MoCA groups. Regarding the heterogeneity of the feature space, we combined the AdaBoost with the Naive Bayes classification algorithm as the decision model of MG. For the screening tool to be used repeatedly, the retrieval practice effect was considered in the design. Phase III involved the validation of measuring instruments. The features incorporated into the modeling process were optimized based on the classification accuracy and area under curve. We also verified the classification effect of the other three classification models with MG. RESULTS: After three steps of feature engineering, a total of 6 dimensions of cognitive areas were included in MG: orientation, memory, attention, calculation, recall, and language & executive function. 38 features were included in the model (17 features of option score, 20 features of time score, and 1 demographic feature). A total of 333 individuals from two communities in Shanghai and Henan province were included in the measuring instrument verification process. Women accounted for 68.2% of the sample. The median age was 63. 15.3% of the participants had bachelor's degrees or above and 111 participants lived in urban areas (33.3%). The results showed that MG had an accuracy of 93.75% and AUC of 0.923, with a sensitivity of 91.67% and a specificity of 95.45%. Compared to the other three classification models, MG that combined the AdaBoost with the Naive Bayes classification algorithm was the most accurate classifier. CONCLUSIONS: MG was proved to be reliable and valid in early screening for patients with MCI. MG that integrated heterogeneous features such as demography, option scores, and time scores had a better predictive performance for screening MCI.

Psoralidin protects against cerebral hypoxia/reoxygenation injury: Role of GAS6/Axl signaling.

Psoralidin (PSO) is a natural phenolic coumarin extracted from the seeds of Psoralea corylifolia L. Growing preclinical evidence indicates that PSO has anti-inflammatory, anti-vitiligo, anti-bacterial, and anti-viral effects. Growth arrest-specific gene 6 (GAS6) and its receptor, Axl, modulate cellular oxidative stress, apoptosis, survival, proliferation, migration, and mitogenesis. Notably, the neuroprotective role of the GAS6/Axl axis has been identified in previous studies. We hypothesize that PSO ameliorates cerebral hypoxia/reoxygenation (HR) injury via activating the GAS6/Axl signaling. We first confirmed that PSO was not toxic to the cells and upregulated GAS6 and Axl expression after HR injury. Moreover, PSO exerted a marked neuroprotective effect against HR injury, represented by restored cell viability and cell morphology, decreased lactate dehydrogenase (LDH) release, and reactive oxygen species (ROS) generation. Furthermore, PSO pretreatment also elevated the levels of nuclear factor-related factor 2 (Nrf-2), NAD(P)H dehydrogenase quinone-1 (NQO1), heme oxygenase-1 (HO-1), silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor coactivator 1α (PGC-1α), nuclear respiratory factor 1 (NRF1), uncoupling protein 2 (UCP2), and B-cell lymphoma 2 (BCl2) both in the condition of baseline and HR injury. However, GAS6 siRNA or Axl siRNA inhibited the neuroprotective effects of PSO. Our findings suggest that PSO pretreatment attenuated HR-induced oxidative stress, apoptosis, and mitochondrial dysfunction in neuroblastoma cells through the activation of GAS6/Axl signaling.

Enhanced dielectric and conductivity properties of carbon-coated SiC nanocomposites in the terahertz frequency range.

C-coated SiC nanocomposites (SiC@C NCs) were one-step synthesized under a mixture atmosphere of Ar and CH4 using a DC arc-discharge plasma method. The microstructure of the composites could be controlled by varying the volume ratio of Ar and CH4. A strong response to the terahertz (THz) field was observed due to the existence of a graphite shell. The dielectric properties of SiC@C NCs can be enhanced by altering the thickness of the graphite shell. The thicker graphite shell results in a stronger absorption of THz waves and an enhanced real part of conductivity. Fitting the measured conductivity data using the Drude-Smith model reveals that the carrier transport in the SiC@C NCs and its counterpart, the SiC nanoparticles, is dominated by backscattering. The SiC@C NCs with enhanced conductivity are believed to be fundamental materials for various functionalized optoelectronic devices.

Laser ablation of pristine Fe foil for constructing a layer-by-layer SiO2/Fe2O3/Fe integrated anode for high cycling-stability lithium-ion batteries.

In this paper, an integrated SiO2/Fe2O3/Fe anode is fabricated by straightforward laser ablation of the surface of Fe foil in air. The oxidized surface is subsequently coated with tetraethyl orthosilicate (TEOS) and transformed into a SiO2 layer through a calcination process in an argon atmosphere. The surface oxidation is traced by on-line optical emission spectroscopy (OES) diagnosis. With high electron temperature (∼5200 K) in the laser irradiation zone, the nanostructured Fe2O3 layer is formed on the Fe foil, resulting in the pristine Fe2O3/Fe anode. This greatly simplified procedure with respect to the conventional route allows direct connection between the Fe2O3 layer and the Fe substrate (current collector) without any binder or conductive agent. In addition, the SiO2 coating layer greatly improves the cycling stability due to the compensatory contribution to capacity during the cycling process and its compatible elasticity to accommodate the volume expansion of Fe2O3, which is verified by first-principles theoretical calculations. The integrated SiO2/Fe2O3/Fe anode delivers a stable capacity of 651.7 mA h g-1 at 0.2 A g-1 after 100 cycles. This strategy offers a low-cost route for the rapid fabrication of integrated electrodes, broadening their applications in high cycling-stability LIBs.

Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1.

Immune checkpoint inhibition is an important strategy in cancer therapy. Blockade of CTLA-4 and PD-1/PD-L1 is well developed in clinical practice. In the last few years, LAG-3 has received much interest as an emerging novel target in immunotherapy. It was recently reported that FGL1 is a major ligand of LAG-3, which is normally secreted by the liver but is upregulated in several human cancers. FGL1 is a crucial biomarker and target for cancer immunotherapy. As the efficacy of immunotherapy is limited to specific types of patients, the subset of patients needs to be selected appropriately to receive precise treatment according to different biomarkers. To date, there is no test to accurately assess FGL1 expression levels. Nanobodies have some outstanding features, such as high stability, solubility and affinity for diagnostic and therapeutic applications. Here, we report the development and validation of a rapid, sensitive, and cost-effective nanobody-based immunoassay for the detection of FGL1 in human serum. In this study, human FGL1 recombinant protein was expressed and purified for the first time as an immunized antigen. Then, we constructed a nanobody phage display library and screened several nanobodies that bind FGL1 with high affinity. We selected two nanobodies targeting different epitopes of FGL1, one as a capture and the other conjugated with HRP as a probe. The double nanobody-based sandwich ELISA to detect the concentration of FGL1 showed a good response relationship in the range of 15.625-2000 ng/mL, and the recoveries from the spiked sample were in the range of 78% and 100%. This assay could be used as a potential approach for evaluating FGL1 expression for patient stratification and for predicting the therapeutic efficacy of targeting the LAG3/FGL1 axis.

Identification of GH17 gene family in Vitis vinifera and expression analysis of GH17 under various adversities.

Glycoside hydrolase (GH, EC 3.2.1) is a group of enzymes that hydrolyzes glycosidic bonds and play a role in the hydrolysis and synthesis of sugars in living organisms. Vitis vinifera is an important fruit crop and it harbors GH17 gene family however, their function in grapes has not been systematically investigated. In this study, a total of 870 GH17 genes were identified from 14 plant species and their structural domain, sequence alignment, phylogenetic tree, collinear analysis, with the expression profiles of VviGH17 gene family was performed. The promoter analysis of VviGH17 gene showed the presence of cis-acting elements, which are responsive to plant growth and development. In addition, elements for plant hormones were found that are triggered in response to abiotic/biological stress. Transcriptomic data led to the identification of several VviGH17 genes, which are associated with bud dormancy and in response to abiotic stress. Transcript analysis was carried out for some of the selected VviGH17 genes RT-qPCR. VviGH17-16 and VviGH17-30 genes were differentially expressed during bud dormancy, fruit development and different abiotic stresses. Moreover, VviGH17-37 and VviGH17-44 were differentially expressed at fruit development, in response to abiotic stress. In addition, subcellular localization predicts that the VviGH17-16, VviGH17-30, and VviGH17-37 genes were located in the cell membrane, while VviGH17-44 gene was located in the vacuole. In conclusion, our study led to the identification of several GH17s and their probable role in development and stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01014-1.

Dynamics of fungal community during silage fermentation of elephant grass (Pennisetum purpureum) produced in northern Vietnam.

OBJECTIVE: This study aimed to gain deeper insights into the dynamic changes in spoilage fungi populations during fermentation and the influence of traditional additives on silage quality. METHODS: Elephant grass (Pennisetum purpureum) was prepared without any additive (control), and with the addition of 0.5% salt, and 0.5% salt-0.2% sugar mixture. The fungal community was then determined using a classic culturing method and high-throughput sequencing at 0, 5, 15, and 60 days after ensiling. RESULTS: The results showed that the fungal community of elephant grass silage varied significantly between the natural fermentation without any additive and the two additive groups. The diversity and relative abundance of spoilage molds in the control group were much higher than those in the two treatment groups (p<0.05). Three species of yeasts (Candida sp., Pichia sp., Trichosporon sp.) and four spoilage molds (Fusarium sp., Aspergillus sp., Muco sp. and Penicillin sp.) were the predominant fungi in elephant grass during natural fermentation from 0 to 60 days, which were found to be significantly decreased in salt and sugar additive groups (p<0.05). Meanwhile, the diversity and relative abundance of undesirable molds in the 0.5%-salt additive group were the lowest among all groups. CONCLUSION: Adding salt and sugar, particularly 0.5% salt, is a promising effective approach to reduce the amount of undesirable fungi thus, improving the silage quality of elephant grass in northern Vietnam.

Transcriptome analysis reveals the molecular mechanisms of the defense response to gray leaf spot disease in maize.

BACKGROUND: Gray leaf spot (GLS), which is caused by the necrotrophic fungi Cercospora zeae-maydis and Cercospora zeina, is one of the most impactful diseases in maize worldwide. The aim of the present study is to identify the resistance genes and understand the molecular mechanisms for GLS resistance. RESULTS: Two cultivars, 'Yayu889' and 'Zhenghong532,' which are distinguished as resistant and susceptible cultivars, respectively, were challenged with the GLS disease and a RNA-seq experiment was conducted on infected plants at 81, 89, 91, and 93 days post planting (dap). Compared with the beginning stage at 81 dap, 4666, 1733, and 1166 differentially expressed genes (DEGs) were identified at 89, 91, and 93 dap, respectively, in 'Yayu889,' while relatively fewer, i.e., 4713, 881, and 722 DEGs, were identified in 'Zhenghong532.' Multiple pathways involved in the response of maize to GLS, including 'response to salicylic acid,' 'protein phosphorylation,' 'oxidation-reduction process,' and 'carotenoid biosynthetic process,' were enriched by combining differential expression analysis and Weighted Gene Co-expression Network Analysis (WGCNA). The expression of 12 candidate resistance proteins in these pathways were quantified by the multiple reaction monitoring (MRM) method. This approach identified two candidate resistance proteins, a calmodulin-like protein and a leucine-rich repeat receptor-like protein kinase with SNPs that were located in QTL regions for GLS resistance. Metabolic analysis showed that, compared with 'Zhenghong532,' the amount of salicylic acid (SA) and total carotenoids in 'Yayu889' increased, while peroxidase activity decreased during the early infection stages, suggesting that increased levels of SA, carotenoids, and reactive oxygen species (ROS) may enhance the defense response of 'Yayu889' to GLS. CONCLUSION: By combining transcriptome and proteome analyses with comparisons of resistance QTL regions, calmodulin-like protein and leucine-rich repeat receptor-like protein kinase were identified as candidate GLS resistance proteins. Moreover, we found that the metabolic pathways for ROS, SA, and carotenoids are especially active in the resistant cultivar. These findings could lead to a better understanding of the GLS resistance mechanisms and facilitate the breeding of GLS-resistant maize cultivars.

Detecting and identifying pathogens and antagonistic bacteria associated with Ginkgo biloba leaf spot disease.

Background: Leaf spot disease severely impacts Ginkgo biloba (G. biloba) yield and quality. While microbial agents offer effective and non-toxic biological control for plant diseases, research on controlling leaf spot disease in G. biloba is notably scarce. Methods: The pathogenic fungi were isolated and purified from diseased and healthy leaves of G. biloba, Subsequent examinations included morphological observations and molecular identification via PCR techniques. A phylogenetic tree was constructed to facilitate the analysis of these pathogenic fungi, and Koch's postulates were subsequently employed to reaffirm their pathogenic nature. The antagonistic experiment was employed to select biocontrol bacteria, and subsequently, the isolated biocontrol bacteria and pathogenic fungi were inoculated onto healthy leaves to assess the inhibitory effects of the biocontrol bacteria. Results: Two pathologies responsible for the leaf spot disease on G. biloba were identified as Botryosphaeria dothidea and Neofusicoccum parvum via the analysis of phylogenetic tree and the application of Koch's Postulates. Additionally, we isolated two strains of biocontrol bacteria, namely Bacillus velezensis and Bacillus amyloliquefaciens. Their average inhibitory zones were measured at 4.78 cm and 3.46 cm, respectively. The inhibition zone of B. velezensis against N. parvum was 4 cm. B. velezensis showed a stronger inhibitory effect compared to B. amyloliquefaciens on the development of lesions caused by B. dothidea via leaf culture experiment. Conclusion: This research reports, for the first time, the presence of B. dothidea as a pathogenic fungus affecting G. biloba. Moreover, the biocontrol bacteria, B. velezensis and B. amyloliquefaciens, exhibited the capability to effectively inhibit the growth and reproduction of B. dothidea, indicating their promising potential as environmentally friendly biocontrol resources.

Impact of ultra-pulse carbon dioxide laser on the treatment of chronic refractory ulcer with bone exposure.

BACKGROUND: Chronic refractory ulcers with bone exposure present significant challenges in wound management and necessitate effective treatment strategies to facilitate healing and alleviate patient discomfort. This study aimed to investigate the impact of ultra-pulse carbon dioxide laser on treating chronic refractory ulcers with bone exposure. METHODS: This retrospective observational study enrolled patients diagnosed with chronic refractory ulcers with bone exposure admitted to the wound repair clinic of the Affiliated Hospital of Southwest Medical University between July 2018 and July 2019. RESULTS: A total of 64 patients with chronic refractory ulcers and bone exposure were included, of which 32 patients underwent ultra-pulse carbon dioxide laser drilling. Compared with patients who did not receive ultra-pulse carbon dioxide laser treatment, those who experienced the procedure demonstrated significantly higher wound healing rates on the fourth, eighth, 12th, 16th, and 20th days after treatment (all P < .001), lower scores on the visual analog scale for pain after 20 days of debridement (0.24 ± 0.05 vs 0.58 ± 0.12, P < .001), lower granulation color observation scores on the 12th, 16th, and 20th days (all P = .001), as well as reduced treatment costs (8200 ± 1600 yuan vs 15400 ± 3800 yuan, P < .001). CONCLUSION: Ultra-pulse carbon dioxide laser treatment may enhance the growth of granulation tissue, improve wound healing rates, reduce pain, and lower treatment costs for patients with chronic bone exposure wounds compared to those without such treatment.

Mental health disparities between physicians and nurses: Analyzing the impact of occupational stress and work environment fitness using random forest algorithm.

BACKGROUND: The impact of occupational stress and work environment fitness on mental health disparities between physicians and nurses are not well understood. This study aims to identify and rank key determinants of mental health in physicians and nurses in China and compare the differences in their impact on mental health between physicians and nurses. METHODS: A large cross-sectional survey with multistage cluster sampling was conducted. The survey included the Self-Rating Anxiety Scale (SAS Scale), the Center for Epidemiologic Studies Depression Scale (CES-D Scale), the Maslach Burnout Inventory-General Survey (MBI-GS) and the Person-Environment (PE) Fit. We applied a principled, machine learning-based variable selection algorithm, using random forests, to identify and rank the determinants of the mental health in physicians and nurses. RESULTS: In our study, we analyzed a sample of 9964 healthcare workers, and 2729 (27 %) were physicians. The prevalence of anxiety and depressive disorders among physicians and nurses was 31.0 % and 53.3 %, 30.8 % and 47.9 %, respectively. Among physicians with anxiety disorder, we observed a higher likelihood of cynicism, emotional exhaustion, reduced personal accomplishment, and poor organization fitness, job fitness, group fitness, and supervisor fitness, in order of importance. When comparing the effects on depressive disorder in physicians, group fitness and supervisor fitness did not have significant impacts. For nurses, emotional exhaustion had a more significant effect on depressive disorder compared to cynicism. Supervisor fitness did not have a significant impact on anxiety disorder in nurses. LIMITATIONS: Cross-sectional design, self-reporting screening scales. CONCLUSIONS: Compared to individual and hospital characteristics, the primary factors influencing mental health disorders are occupational burnout and the compatibility of the work environment. Additionally, the key determinants of depressive and anxiety disorders among doctors and nurses exhibit slight variations. Employing machine learning methods proves beneficial for identifying determinants of mental health disorders among physicians and nurses in China. These findings could help improve policymaking aimed at addressing the mental well-being of healthcare professionals.

Single-Cell Transcriptome Landscape and Cell Fate Decoding in Human Brain Organoids after Transplantation.

Human stem cells and derivatives transplantation are widely used to treat nervous system diseases, while the fate determination of transplanted cells is not well elucidated. To explore cell fate changes of human brain organoids before and after transplantation, human brain organoids are transplanted into prefrontal cortex (PFC) and hippocampus (HIP), respectively. Single-cell sequencing is then performed. According to time-series sample comparison, transplanted cells mainly undergo neural development at 2 months post-transplantation (MPT) and then glial development at 4MPT, respectively. A different brain region sample comparison shows that organoids grafted to PFC have obtained cell fate close to those of host cells in PFC, other than HIP, which may be regulated by the abundant expression of dopamine (DA) and acetylcholine (Ach) in PFC. Meanwhile, morphological complexity of human astrocyte grafts is greater in PFC than in HIP. DA and Ach both activate the calcium activity and increase morphological complexity of astrocytes in vitro. This study demonstrates that human brain organoids receive host niche factor regulation after transplantation, resulting in the alignment of grafted cell fate with implanted brain regions, which may contribute to a better understanding of cell transplantation and regenerative medicine.

Health care costs of cardiovascular disease in China: a machine learning-based cross-sectional study.

Background: Cardiovascular disease (CVD) causes substantial financial burden to patients with the condition, their households, and the healthcare system in China. Health care costs for treating patients with CVD vary significantly, but little is known about the factors associated with the cost variation. This study aims to identify and rank key determinants of health care costs in patients with CVD in China and to assess their effects on health care costs. Methods: Data were from a survey of patients with CVD from 14 large tertiary grade-A general hospitals in S City, China, between 2018 and 2020. The survey included information on demographic characteristics, health conditions and comorbidities, medical service utilization, and health care costs. We used re-centered influence function regression to examine health care cost concentration, decomposing and estimating the effects of relevant factors on the distribution of costs. We also applied quantile regression forests-a machine learning approach-to identify the key factors for predicting the 10th (low), 50th (median), and 90th (high) quantiles of health care costs associated with CVD treatment. Results: Our sample included 28,213 patients with CVD. The 10th, 50th and 90th quantiles of health care cost for patients with CVD were 6,103 CNY, 18,105 CNY, and 98,637 CNY, respectively. Patients with high health care costs were more likely to be older, male, and have a longer length of hospital stay, more comorbidities, more complex medical procedures, and emergency admissions. Higher health care costs were also associated with specific CVD types such as cardiomyopathy, heart failure, and stroke. Conclusion: Machine learning methods are useful tools to identify determinants of health care costs for patients with CVD in China. Findings may help improve policymaking to alleviate the financial burden of CVD, particularly among patients with high health care costs.

Refinement of CHADS2 and CHA2DS2-VASc scores predict left atrial thrombus or spontaneous echo contrast in nonvalvular atrial fibrillation patients.

OBJECTIVE: To investigate the risk factors of left atrial thrombus (LAT)/spontaneous echo contrast (SEC) in patients with nonvalvular atrial fibrillation (AF). METHODS: This retrospective study analysed the data from consecutive patients with nonvalvular AF that underwent transoesophageal echocardiography. Logistic regression analysis was performed to identify risk factors of LAT/SEC. Receiver operating characteristic curve analysis was undertaken compare the new scales with CHADS2 and CHA2DS2-VASc scores. RESULTS: A total of 558 patients with AF were included in the study. LAT/SEC was detected in 137 (24.6%) patients. The independent risk factors of LAT/SEC beyond CHADS2 or CHA2DS2-VASc scores included non-paroxysmal AF and left atrial diameter >37.5 mm. These two variables were added into the CHADS2 or CHA2DS2-VASc score to build new scales. Areas under the curve for the new scales based on CHADS2 and CHA2DS2-VASc scores were significantly higher than the CHADS2 or CHA2DS2-VASc score both in the overall study cohort and in patients at a high risk of thromboembolism. CONCLUSIONS: Non-paroxysmal AF and increased left atrial diameter beyond the CHADS2 or CHA2DS2-VASc score were independent risk factors of LAT/SEC and may help to improve the current risk stratification, especially for patients with nonvalvular AF at a high risk of thromboembolism.

Lycorine and organ protection: Review of its potential effects and molecular mechanisms.

BACKGROUND: Multiorgan dysfunction, especially sepsis-related multiorgan damage, remains a major cause of high mortality in the late stages of infection and a great clinical challenge. In recent years, natural drugs have received widespread attention because of their low cost, wide sources, high efficacy, low toxicity, and limited side effects. Lycorine, a natural compound extracted from Amaryllidaceae, exhibits multiple pharmacological activities, including in the regulation of autophagy and the induction of cancer cell apoptosis, and has anti-inflammatory, antifungal, antiviral, antimalarial, and antitumor activities. However, studies on lycorine have mainly focused on its antitumor properties, and research on its use for organ protection, especially in sepsis-related organ injury, is relatively limited. PURPOSE: To review and discuss the effects and mechanisms of lycorine in the treatment of multi-organ dysfunction, especially sepsis. METHODS: Literature searches in electronic databases, such as Web of Science, Science Direct, PubMed, Google Scholar, and Scopus, were performed using 'Lycorine', 'Amaryllidaceae', 'Pharmacology', 'Pharmacokinetics', 'Anti-inflammation', 'Autophagy', 'Apoptosis', 'Anti-microbial and anti-parasitic', 'Antitumor', 'Organ protection', and 'Sepsis' as keywords, the correlated literature was extracted and conducted from the databases mentioned above. RESULTS: By summarizing the progress made in existing research, we found that the general effects of lycorine involve the regulation of autophagy and the induction of cancer cell apoptosis, and anti-inflammatory, antifungal, antiviral, antimalarial, and antitumor effects; through these pathways, the compound can ameliorate organ damage. In addition, lycorine was found to have an important effect on organ damage in sepsis. CONCLUSION: Lycorine is a promising natural organ protective agent. This review will provide a new theoretical basis for the treatment of organ protection, especially in sepsis.