A genomic mutational constraint map using variation in 76,156 human genomes.

The depletion of disruptive variation caused by purifying natural selection (constraint) has been widely used to investigate protein-coding genes underlying human disorders1-4, but attempts to assess constraint for non-protein-coding regions have proved more difficult. Here we aggregate, process and release a dataset of 76,156 human genomes from the Genome Aggregation Database (gnomAD)-the largest public open-access human genome allele frequency reference dataset-and use it to build a genomic constraint map for the whole genome (genomic non-coding constraint of haploinsufficient variation (Gnocchi)). We present a refined mutational model that incorporates local sequence context and regional genomic features to detect depletions of variation. As expected, the average constraint for protein-coding sequences is stronger than that for non-coding regions. Within the non-coding genome, constrained regions are enriched for known regulatory elements and variants that are implicated in complex human diseases and traits, facilitating the triangulation of biological annotation, disease association and natural selection to non-coding DNA analysis. More constrained regulatory elements tend to regulate more constrained protein-coding genes, which in turn suggests that non-coding constraint can aid the identification of constrained genes that are as yet unrecognized by current gene constraint metrics. We demonstrate that this genome-wide constraint map improves the identification and interpretation of functional human genetic variation.

DOCK2 is involved in the host genetics and biology of severe COVID-19.

Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge1-5. Here we conducted a genome-wide association study (GWAS) involving 2,393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3,289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target.

The mutational constraint spectrum quantified from variation in 141,456 humans.

Genetic variants that inactivate protein-coding genes are a powerful source of information about the phenotypic consequences of gene disruption: genes that are crucial for the function of an organism will be depleted of such variants in natural populations, whereas non-essential genes will tolerate their accumulation. However, predicted loss-of-function variants are enriched for annotation errors, and tend to be found at extremely low frequencies, so their analysis requires careful variant annotation and very large sample sizes1. Here we describe the aggregation of 125,748 exomes and 15,708 genomes from human sequencing studies into the Genome Aggregation Database (gnomAD). We identify 443,769 high-confidence predicted loss-of-function variants in this cohort after filtering for artefacts caused by sequencing and annotation errors. Using an improved model of human mutation rates, we classify human protein-coding genes along a spectrum that represents tolerance to inactivation, validate this classification using data from model organisms and engineered human cells, and show that it can be used to improve the power of gene discovery for both common and rare diseases.

VEGF-A in serum protects against memory impairment in APP/PS1 transgenic mice by blocking neutrophil infiltration.

Activation of innate immunity in the brain is a prominent feature of Alzheimer's disease (AD). The present study investigated the regulation of innate immunity by wild-type serum injection in a transgenic AD mouse model. We found that treatment with wild-type mouse serum significantly reduced the number of neutrophils and microglial reactivity in the brains of APP/PS1 mice. Mimicking this effect, neutrophil depletion via Ly6G neutralizing antibodies resulted in improvements in AD brain functions. Serum proteomic analysis identified vascular endothelial growth factor-A (VEGF-A) and chemokine (C-X-C motif) ligand 1 (CXCL1) as factors enriched in serum samples, which are crucial for neutrophil migration and chemotaxis, leukocyte migration, and cell chemotaxis. Exogenous VEGF-A reversed amyloid ß (Aß)-induced decreases in cyclin-dependent kinase 5 (Cdk5) and increases in CXCL1 in vitro and blocked neutrophil infiltration into the AD brain. Endothelial Cdk5 overexpression conferred an inhibitory effect on CXCL1 and neutrophil infiltration, thereby restoring memory abilities in APP/PS1 mice. Our findings uncover a previously unknown link between blood-derived VEGF signaling and neutrophil infiltration and support targeting endothelial Cdk5 signaling as a potential therapeutic strategy for AD.

Analysis of health service utilization and influencing factors due to COVID-19 in Beijing: a large cross-sectional survey.

BACKGROUND: In the wake of China's relaxed zero-COVID policy, there was a surge in coronavirus disease 2019 (COVID-19) infections. This study aimed to examine the infection status and health service utilization among Beijing residents during a widespread outbreak, and to explore the factors that affected utilization of health services due to COVID-19. METHODS: A cross-sectional survey was conducted among Beijing residents from 13 January to 13 February 2023, collecting information on socio-demographic characteristics, health behaviours, COVID-19 infection status, utilization of health services and depressive symptoms. Multivariate Tobit regression was used for data analysis. RESULTS: Among the 53 924 participants, 14.7% were older than 60 years, 63.7% were female and 84.8% were married. In total, 44 992 of the 53 924 individuals surveyed (83.4%) contracted COVID-19 during 2020-2023, and 25.2% (13 587) sought corresponding health services. The majority of individuals (85.6%) chose in-person healthcare, while 14.4% chose internet-based healthcare. Among those who chose in-person healthcare, 58.6% preferred primary healthcare institutions and 41.5% were very satisfied with the treatment. Factors affecting health service utilization include being female (ß = -0.15, P < 0.001), older than 60 years (ß = 0.23, P < 0.01), non-healthcare workers (ß = -0.60, P < 0.001), rich self-rated income level (ß = 0.59, P < 0.001), having underlying disease (ß = 0.51, P < 0.001), living alone (ß = -0.19, P < 0.05), depressive symptoms (ß = 0.06, P < 0.001) and healthy lifestyle habits, as well as longer infection duration, higher infection numbers and severe symptoms. CONCLUSION: As COVID-19 is becoming more frequent and less severe, providing safe and accessible healthcare remains critical. Vulnerable groups such as the elderly and those with underlying conditions need reliable health service. Prioritizing primary healthcare resources and online medical services have played a vital role in enhancing resource utilization efficiency.

Middle Meningeal Arterial Embolization Combined With Drilling in the Treatment of Acute Epidural Hematoma.

OBJECTIVE: The purpose of this study was to determine the technical feasibility and safety of middle meningeal arterial (MMA) embolization combined with drilling drainage in the treatment of acute epidural hematoma (AEDH) by comparing it with traditional craniotomy in the treatment. METHODS: One hundred seventeen patients with AEDH treated for MMA embolization combined with drilling and drainage or craniotomy hematoma removal from January 2017 to September 2020 were retrospectively analyzed and divided into a craniotomy group (n=85) and a minimally invasive group (n=32). Hematoma removal was performed in the craniotomy group, and MMA embolization combined with drilling and drainage was performed in the minimally invasive group. The general clinical data, imaging data, surgery, and follow-up of the 2 groups were compared and analyzed. RESULTS: Compared with the craniotomy group, the residual hematoma volume in the minimally invasive group was higher than in the craniotomy group. The average postoperative drainage duration in the minimally invasive group was longer than in the craniotomy group. Compared with the craniotomy group, the minimally invasive group was associated with shorter operative time, less intraoperative bleeding, and lower rates of postoperative rebleeding. In addition, the incidence of postoperative complications and length of hospitalization in the minimally invasive group were significantly shortened. CONCLUSION: Middle meningeal arterial embolization combined with drilling and drainage in the treatment of AEDH caused by MMA active bleeding is safe, effective, and more minimally invasive, and can be promoted and applied.

Research on the Technological Progress of CZT Array Detectors.

CdZnTe (CZT) is a new type of compound semiconductor that has emerged in recent years. Compared to other semiconductor materials, it possesses an ideal bandgap, high density, and high electron mobility, rendering it an excellent room-temperature composite semiconductor material for X-ray and γ-ray detectors. Due to the exceptional performance of CZT material, detectors manufactured using it exhibit high energy resolution, spatial resolution, and detection efficiency. They also have the advantage of operating at room temperature. CZT array detectors, furthermore, demonstrate outstanding spatial detection and three-dimensional imaging capabilities. Researchers worldwide have conducted extensive studies on this subject. This paper, building upon this foundation, provides a comprehensive analysis of CZT crystals and CZT array detectors and summarizes existing research to offer valuable insights for envisioning new detector methodologies.

[Main characteristics and historical evolution of China' s health financing transition].

OBJECTIVE: To comprehend the main characteristics and historical evolution of health financing transition in China. METHODS: Data were collected from various sources, including the Global Health Expenditure Database (GHED), China Health Statistics Yearbook, National Health Finance Annual Report, China ' s Total Health Expenditure Research Report, et al. Descriptive statistics and literature study was conducted. RESULTS: Since the beginning of the 21st century, most countries in the world had witnessed a transition of health financing, characterized by the expansion of health financing scale and the strengthening of public financing responsibility. Notably, China ' s health financing transition exhibited distinctive features. Firstly, there had been a more rapid expansion in health financing scale compared with global averages. Between 2000 and 2019, total health expenditure per capita experienced a remarkable increase of 816.6% at comparable prices, significantly surpassing average growth rates observed among other countries worldwide (102.1%). Secondly, greater efforts had been made to strengthen the responsibilities of public financing. From 2000 to 2019, there was a substantial decrease of 30.6 percentage points in the proportion of out-of-pocket health expenditure as a share of total health expenditure. This decline was significantly larger than the average reduction observed among other countries worldwide (5.6 percentage points). Thirdly, there had been a significant shift in government health expenditure allocation patterns, with an increased emphasis on "demand-side subsidies" surpassing "supply-side subsidies". Within the realm of "supply-side subsidies", funding directed towards hospitals had notably increased and surpassed that allocated to primary healthcare institutions and public health institutions. Based on these distinctive characteristics, this paper expanded China ' s health financing transition into three dimensions: Scale dimension, structure dimension and flow dimension. Using a comprehensive analytical framework, the history of China ' s health financing transition was roughly divided into four stages: The planned economy stage, the economic transition stage, the post-SARS stage and the new health system reform stage. The main features and evolutionary logic associated with each stage were analyzed. CONCLUSION: Above all, the health financing system should be enhanced in terms of vertical "embeddedness" and horizontal "complementarity". Moreover, the significance of health financing transition in preserving hidden value and mitigating public risk should be emphasized, and there is a need for an improved two-way trade-off mechanism that balances value and risk. Additionally, the ethical principles associated with health financing transition should be considered comprehensively, while optimizing budget decision-making within the government ' s actual governance model. Lastly, it is crucial to recognize the overall and profound impact of modern medicine development and explore long-term strategies and pathways for health financing transition in China.

Template-Directed Polymerization of Binary Acrylate Monomers on Surface-Activated Lignin Nanoparticles in Toughening of Bio-Latex Films.

Fabricating bio-latex colloids with core-shell nanostructure is an effective method for obtaining films with enhanced mechanical characteristics. Nano-sized lignin is rising as a class of sustainable nanomaterials that can be incorporated into latex colloids. Fundamental knowledge of the correlation between surface chemistry of lignin nanoparticles (LNPs) and integration efficiency in latex colloids and from it thermally processed latex films are scarce. Here, an approach to integrate self-assembled nanospheres of allylated lignin as the surface-activated cores in a seeded free-radical emulsion copolymerization of butyl acrylate and methyl methacrylate is proposed. The interfacial-modulating function on allylated LNPs regulates the emulsion polymerization and it successfully produces a multi-energy dissipative latex film structure containing a lignin-dominated core (16% dry weight basis). At an optimized allyl-terminated surface functionality of 1.04 mmol g-1 , the LNPs-integrated latex film exhibits extremely high toughness value above 57.7 MJ m-3 . With multiple morphological and microstructural characterizations, the well-ordered packing of latex colloids under the nanoconfinement of LNPs in the latex films is revealed. It is concluded that the surface chemistry metrics of colloidal cores in terms of the abundance of polymerization-modulating anchors and their accessibility have a delicate control over the structural evolution of core-shell latex colloids.

Observation of the "invisible" pulsation of soliton molecules in a bidirectional ultrafast fiber laser.

A novel optical soliton dynamics phenomenon, called "invisible" pulsation, has gradually attracted extensive interest in recent years, which can only be identified effectively with the help of real-time spectroscopy technique, i.e., dispersive Fourier transformation (DFT). In this paper, based on a new bidirectional passively mode-locked fiber laser (MLFL), the "invisible" pulsation dynamics of soliton molecules (SMs) is systematically studied. It is indicated that the spectral center intensity, pulse peak power and relative phase of SMs are periodically changed during the "invisible" pulsation, while the temporal separation inside the SMs is constant. The degree of spectral distortion is positively correlated with the pulse peak power, which verifies that self-phase modulation (SPM) is the inducement of spectral distortion. Finally, the universality of the SMs "invisible" pulsation is further experimentally verified. We believe our work is not only conducive to the development of compact and reliable bidirectional ultrafast light sources, but also of great significance to enrich the study of nonlinear dynamics.

"Invisible" pulsation and period-doubling bifurcation of harmonic mode locking in a bidirectional fiber laser.

The harmonic mode-locking (HML) "invisible" pulsation (IP) is reported, here, in a bidirectional passively mode-locked fiber laser (BPMLFL). With the help of dispersive Fourier transform (DFT) technology, it is found that due to the alike nonlinear effects experienced by two pulse trains in HML, their evolution is consistent during the IP. Further, as the increase of pump power, period-doubling bifurcations (PDBs) can be observed based on the IP phenomenon in the HML regime, the PDB path experienced by the HML from steady to chaotic is statistically obtained. Finally, the IP and PDB in the bidirectional laser are reproduced and studied through numerical simulations. The effect of IP on the coherence of solitons is further analyzed. We believe our research results will provide new insights into the study of soliton dynamics in fiber lasers.

Aqueous Processable One-Dimensional Polypyrrole Nanostructured by Lignocellulose Nanofibril: A Conductive Interfacing Biomaterial.

One-dimensional (1D) nanomaterials of conductive polypyrrole (PPy) are competitive biomaterials for constructing bioelectronics to interface with biological systems. Synergistic synthesis using lignocellulose nanofibrils (LCNF) as a structural template in chemical oxidation of pyrrole with Fe(III) ions facilitates surface-confined polymerization of pyrrole on the nanofibril surface within a submicrometer- and micrometer-scale fibril length. It yields a core-shell nanocomposite of PPy@LCNF, wherein the surface of each individual fibril is coated with a thin nanoscale layer of PPy. A highly positive surface charge originating from protonated PPy gives this 1D nanomaterial a durable aqueous dispersity. The fibril-fibril entanglement in the PPy@LCNFs facilely supported versatile downstream processing, e.g., spray thin-coating on glass, flexible membranes with robust mechanics, or three-dimensional cryogels. A high electrical conductivity in the magnitude of several to 12 S·cm-1 was confirmed for the solid-form PPy@LCNFs. The PPy@LCNFs are electroactive and show potential cycling capacity, encompassing a large capacitance. Dynamic control of the doping/undoping process by applying an electric field combines electronic and ionic conductivity through the PPy@LCNFs. The low cytotoxicity of the material is confirmed in noncontact cell culture of human dermal fibroblasts. This study underpins the promises for this nanocomposite PPy@LCNF as a smart platform nanomaterial in constructing interfacing bioelectronics.

High-order mode erbium-doped fiber laser based on cavity LPFG converters.

We experimentally investigate high-order mode (HOM) generation at a wavelength of 1.5 µm in an all-fiber erbium-doped laser based on a long-period fiber grating (LPFG). The CW laser emission is achieved when the pump power is above the threshold of 10 mW. An LPFG with a 15 dB bandwidth of 147.76 nm from 1502.76 nm to 1650.52 nm is used as a mode converter inside the cavity. The generation of the broadband L P 11 mode is ultimately obtained. By using a few-mode output coupler, we can obtain the intracavity conversion of the linear polarization mode. Single-, dual-, triple-, and quadruple-wavelength operations can be achieved by changing the polarization state of the polarization controllers in the cavity. The tunable range of the output wavelengths is up to ∼23.20n m. The output power and slope efficiency of the HOMs are presented and discussed. We believe our work might benefit the investigation of HOM fiber lasers, and might be further applied to the intracavity conversion of the linear polarization mode or orbital angular momentum beams.

HyPR-seq: Single-cell quantification of chosen RNAs via hybridization and sequencing of DNA probes.

Single-cell quantification of RNAs is important for understanding cellular heterogeneity and gene regulation, yet current approaches suffer from low sensitivity for individual transcripts, limiting their utility for many applications. Here we present Hybridization of Probes to RNA for sequencing (HyPR-seq), a method to sensitively quantify the expression of hundreds of chosen genes in single cells. HyPR-seq involves hybridizing DNA probes to RNA, distributing cells into nanoliter droplets, amplifying the probes with PCR, and sequencing the amplicons to quantify the expression of chosen genes. HyPR-seq achieves high sensitivity for individual transcripts, detects nonpolyadenylated and low-abundance transcripts, and can profile more than 100,000 single cells. We demonstrate how HyPR-seq can profile the effects of CRISPR perturbations in pooled screens, detect time-resolved changes in gene expression via measurements of gene introns, and detect rare transcripts and quantify cell-type frequencies in tissue using low-abundance marker genes. By directing sequencing power to genes of interest and sensitively quantifying individual transcripts, HyPR-seq reduces costs by up to 100-fold compared to whole-transcriptome single-cell RNA-sequencing, making HyPR-seq a powerful method for targeted RNA profiling in single cells.

Aggravation of nitrogen losses driven by agriculture and livestock farming development on the Qinghai-Tibet Plateau.

Nitrogen (N) losses from crop-livestock production is a major threat to the environment and human health at regional, national and global scales. A comprehensive understanding of the sources, spatiotemporal distribution and drivers of N losses is of great significance for mitigating its negative impacts and promoting N sustainable management. Here, we used the county-scale N flow model to quantitatively analyze the N losses and their driving forces of crop-livestock production on the Qinghai-Tibet Plateau (QTP). Between 2000 and 2018, the total N losses increased for more than 79% of counties on the QTP. The hotspot areas accounted for over 80% of total N losses, expanding from the east and south to the north and west of the QTP. NH3 was the main source of atmospheric N losses (over 80%) while the direct discharge of manure was the main source of water N losses. Structural equation modeling (SEM) showed that chemical fertilizer caused the largest driving effect on atmospheric N losses, and the total output value of agriculture and forestry was the main driver of water N losses. Uneven distribution of crop production and livestock contributed to the aggravation of N losses. Over 70% of counties had grater manure N excretion than crops could take up, and large proportion of manure could not be returned to the field. More than 90% of the counties used grater amount of chemical fertilizer N than crops could take up, indicating that livestock manure has not yet fully replaced chemical fertilizer N. The results provide effective guidance and support for N utilization and management of livestock in agricultural and pastoral areas.

Identification of key priority areas under different ecological restoration scenarios on the Qinghai-Tibet Plateau.

With the intensification of climate warming and human activities, the ecosystems on the Qinghai-Tibet Plateau (QTP) are facing increasing threats which leads to extensive ecological degradation. Ecological restoration measures need to be implemented to improve biodiversity and ecosystem services to mitigate the impact of climate change and human disturbances. However, the key priority areas (KPAs) for ecological restoration are not clear on the QTP, and the benefits of ecosystem services for ecological restoration are often ignored. In this study, we are the first to identify the KPAs based on the quantitative evaluation method and multicriteria optimization algorithm under five restoration scenarios aiming at ecosystem service improvement on the QTP. Results showed that: (1) The benefits of ecological restoration for climate change mitigation and associated costs under different scenarios showed generally similar spatial variability, exhibiting higher in the south and lower in the west, which were different from those for biodiversity. (2) The restoration priorities in Sichuan and Yunnan were generally higher under scenarios Ⅱ and Ⅴ, while in Xinjiang, Sichuan, Yunnan, and western and southern Tibet were higher under scenarios Ⅰ, Ⅲ and Ⅳ. (3) For different ecosystems, the similarities lied in that the restoration priorities of wetland ecosystem were the highest, while those of desert ecosystem were the lowest under five restoration scenarios. (4) When the restoration area requirement was 25% of the total degraded area, the highest restoration priority levels under scenarios Ⅰ, Ⅲ and Ⅳ were mainly distributed in Guinan, Renbu, Nierong and Chayu counties, and under scenarios Ⅱ and Ⅴ were mainly distributed in Renbu, Lang and Guinan counties. When the restoration area requirements were 50% and 75% of the total degraded area, the counties with higher restoration priority levels under scenario Ⅱ were different from those under other four scenarios. This study identified the KPAs under different restoration scenarios, which provided references for the restoration measures implementation on the QTP.