Spatial-temporal drivers and incidence heterogeneity of hemorrhagic fever with renal syndrome transmission in Shandong Province, China, 2016-2022.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) signals a recurring risk in Eurasia in recent years owing to its continued rise in case notifications and the extension of geographical distribution. This study was undertaken to investigate the spatiotemporal drivers and incidence heterogeneity of HFRS transmission in Shandong Province. METHODS: The epidemiological data for HFRS, meteorological data and socioeconomic data were obtained from China Information System for Disease Control and Prevention, China Meteorological Data Sharing Service System, and Shandong Statistical Yearbook, respectively. The spatial-temporal multicomponent model was employed to analyze the values of spatial-temporal components and the heterogeneity of HFRS transmission across distinct regions. RESULTS: The total effect values of the autoregressive, epidemic, and endemic components were 0.451, 0.187, and 0.033, respectively, exhibiting significant heterogeneity across various cities. This suggested a pivotal role of the autoregressive component in propelling HFRS transmission in Shandong Province. The epidemic component of Qingdao, Weifang, Yantai, Weihai, and Jining declined sharply at the onset of 2020. The random effect identified distinct incidence levels associated with Qingdao and Weifang, signifying regional variations in HFRS occurrence. CONCLUSIONS: The autoregressive component emerged as a significant driver in the transmission of HFRS in Shandong Province. Targeted preventive measures should be strategically implemented across various regions, taking into account the predominant component influencing the epidemic.

Discovery of P450-Modified Sesquiterpenoids Levinoids A-D through Global Genome Mining.

Cytochrome P450-modified bacterial terpenoids remain in a vast chemical space to be explored. In the present study, we conducted global genome mining of 223,829 bacterial genomes and identified 2892 bacterial terpenoid biosynthetic gene clusters (BGCs) with cytochrome P450 genes. Among these, we selected 562 with multiple P450 enzymes, which were further clustered as 355 gene cluster families by sequence similarity analysis. We then chose lev, a BGC from Streptomyces levis MCCC1A01616, for heterologous expression and discovered four new α-amorphene-type sesquiterpenoids, levinoids A-D (1-4). The structures and absolute configurations of these four new compounds were determined by employing extensive NMR analysis, NMR chemical shift calculations with DP4+, and ECD calculations. Furthermore, levinoid C (3) exhibited a moderate level of neuroprotective activity (EC50 = 21 µM) in the glutamate-induced excitotoxicity cell model. Our findings highlight the untapped chemical diversity of P450-modified bacterial terpenoids, opening new avenues for further exploration and discovery.

Orally Deliverable Sequence-Targeted Fucoxanthin-Loaded Biomimetic Extracellular Vesicles for Alleviation of Nonalcoholic Fatty Liver Disease.

Extracellular vesicles (EVs) possess favorable biocompatibility and immunological characteristics, making them optimal carriers for bioactive substances. In this study, an innovative hepatic-targeted vesicle system encapsulating with fucoxanthin (GA-LpEVs-FX) was successfully designed and used to alleviate nonalcoholic fatty liver disease. The formulation entails the self-assembly of EVs derived from Lactobacillus paracasei (LpEVs), modification with glycyrrhetinic acid (GA) via amide reaction offering the system liver-targeting capacity and loading fucoxanthin (FX) through sonication treatment. In vitro experiments demonstrated that GA-LpEVs-FX effectively mitigated hepatic lipid accumulation and attenuated reactive oxygen species-induced damage resulting lipid accumulation (p < 0.05). In vivo, GA-LpEVs-FX exhibited significant downregulation of lipogenesis-related proteins, namely, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC1), and sterol regulatory element binding protein 1 (SREBP-1), subsequently ameliorating lipid metabolism disorders (p < 0.05), and the stability of GA-LpEVs-FX significantly improved compared to free FX. These findings establish a novel formulation for utilizing foodborne components for nonalcoholic fatty liver disease alleviation.

Continuous genetic monitoring of transient mesenchymal gene activities in distal tubule and collecting duct epithelial cells during renal fibrosis.

Epithelial cells (ECs) have been proposed to contribute to myofibroblasts or fibroblasts through epithelial-mesenchymal transition (EMT) during renal fibrosis. However, since EMT may occur dynamically, transiently, and reversibly during kidney fibrosis, conventional lineage tracing based on Cre-loxP recombination in renal ECs could hardly capture the transient EMT activity, yielding inconsistent results. Moreover, previous EMT research has primarily focused on renal proximal tubule ECs, with few reports of distal tubules and collecting ducts. Here, we generated dual recombinases-mediated genetic lineage tracing systems for continuous monitoring of transient mesenchymal gene expression in E-cadherin+ and EpCAM+ ECs of distal tubules and collecting ducts during renal fibrosis. Activation of key EMT-inducing transcription factor (EMT-TF) Zeb1 and mesenchymal markers αSMA, vimentin, and N-cadherin, were investigated following unilateral ureteral obstruction (UUO). Our data revealed that E-cadherin+ and EpCAM+ ECs did not transdifferentiate into myofibroblasts, nor transiently expressed these mesenchymal genes during renal fibrosis. In contrast, in vitro a large amount of cultured renal ECs upregulated mesenchymal genes in response to TGF-ß, a major inducer of EMT.

Changes in Cytokine Levels in Patients with Severe Fever with Thrombocytopenia Syndrome Virus.

Purpose: To characterize the cytokine profile of patients with severe fever with thrombocytopenia syndrome (SFTS) in relation to disease severity. Patients and Methods: 60 laboratory-confirmed SFTS patients and 12 healthy individuals from multi-centers in Shandong Province of China were included, and all patients were divided into fatal patients (9) and recovered patients (51) due to their final outcomes. Multiplex-microbead immunoassays were conducted to estimate levels of 27 cytokines in the sera of patients and controls. Results: The results showed that levels of IL-2, IL-4, IL-6, IL-7, IL-8, IL-15, IL-1RA, G-CSF, GM-CSF, IFN-γ, TNF-α, basic FGF, PDGF-BB, RANTES, IP-10, MIP-1α, MIP-1ß, MCP-1, and Eotaxin differed significantly among the SFTS fatal patients, recovered patients, and the healthy controls (all p<0.05). Compared to the healthy controls, the fatal patients and recovered patients had reduced levels of IL-2, IL-4, IL-7, PDGF-BB, RANTES, and Eotaxin, while the levels of PDGF-BB and RANTES were significantly lower in fatal patients compared to recovered patients. The increasing levels of IL-6, IL-8, IL-15, IL-1RA, G-CSF, GM-CSF, IFN-γ, TNF-α, basic FGF, IP-10, MIP-1α, MIP-1ß, and MCP-1 were observed in fatal patients (all p<0.05), and the levels of IL-6, IP-10, MIP-1α, and MCP-1 were significantly higher than other two groups. The Spearman correlation analysis indicated a positive correlation between platelet count and PDGF-BB levels (p<0.05), while the white blood cell count had a negative correlation with MIP-1 level (p<0.05). Conclusion: The research exhibited that the SFTS virus (SFTSV) caused an atypical manifestation of cytokines. The levels of IL-6, IP-10, MIP-1α, and MCP-1 had been observed a positive association with the severity of the illness.

Microfluidic fabrication of core-shell fucoxanthin nanofibers with improved environmental stability for reducing lipid accumulation in vitro.

Fucoxanthin is a xanthophyll carotenoid that possesses potent antioxidant, anti-obesity, and anti-tumor properties. However, its limited solubility in water and susceptibility to degradation create challenges for its application. In this study, a microfluidic coaxial electrospinning technique was used to produce core-shell zein-gelatin nanofibers for encapsulating fucoxanthin, enhancing its bioavailability, and improving its stability. In comparison to uniaxially-loaded fucoxanthin nanofibers, the encapsulation efficiency of fucoxanthin reached 98.58 % at a core-shell flow rate ratio of 0.26:1, representing a 14.29 % improvement. The photostability of the nanofibers increased by 74.59 % after three days, UV stability increased by 38.82 % after 2 h, and temperature stability also significantly improved, demonstrating a protective effect under harsh environmental conditions (P < 0.05). Additionally, nanofibers effectively alleviated oleic acid-induced reactive oxygen species production and reduced fluorescence intensity by 54.76 %. MTT experiments indicated great biocompatibility of the nanofibers, effectively mitigating mitochondrial membrane potential polarization and lipid accumulation in HepG2 cells. Overall, the microfluidic coaxial electrospinning technique enables promising applications of fucoxanthin delivery in the food industry.

Survivability of probiotics in Pickering emulsion gels stabilized by salmon by-product protein / sodium alginate soluble complexes at neutral pH.

Adequate amounts of live probiotics reaching the gut are necessary to maintain host health. However, the harsh environment during processing, the low pH of human gastric acid, and the high concentration of bile salts in the gut can significantly reduce survivability of probiotics. In this work, we propose a simple Pickering emulsion gels strategy to encapsulate Lactobacillus plantarum Lp90 into oil droplets filled in calcium alginate gels to improve its viability under pasteurization and gastrointestinal conditions. The emulsion gels were stabilized by the soluble complexes of salmon by-product protein (SP) and sodium alginate (ALG), and the aqueous phase was solidified by the addition of calcium. The interaction between SP and ALG and the effect of ALG concentration on emulsifying ability and emulsion stability were studied. The results from optical imaging, nuclear magnetic resonance, and rheological properties showed that the stability and viscosity of the emulsions gradually increased with the increased ALG concentration, while the droplet size of the emulsions and the content of free water in the system decreased significantly. Especially when the concentration of ALG was 1 %, the emulsion system was stable under the environment of high temperature and high ionic strength, and the water holding capacity was the highest. Through pasteurization and gastrointestinal digestion experiments, it was found that the survival rate of probiotics encapsulated in emulsion gels was significantly higher than that encapsulated in emulsions or hydrogels, which benefited from the dual action of oil droplets and calcium alginate gels network. These results provide a new strategy for the processing of probiotics and the high-value utilization of marine fish by-products.

Spatiotemporal distribution and environmental influences of severe fever with thrombocytopenia syndrome in Shandong Province, China.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease discovered in China in 2009. The purpose of this study was to describe the spatiotemporal distribution of SFTS and to identify its environmental influencing factors and potential high-risk areas in Shandong Province, China. METHODS: Data on the SFTS incidence from 2010 to 2021 were collected. Spatiotemporal scan statistics were used to identify the time and area of SFTS clustering. The maximum entropy (MaxEnt) model was used to analyse environmental influences and predict high-risk areas. RESULTS: From 2010 to 2021, a total of 5705 cases of SFTS were reported in Shandong. The number of SFTS cases increased yearly, with a peak incidence from April to October each year. Spatiotemporal scan statistics showed the existence of one most likely cluster and two secondary likely clusters in Shandong. The most likely cluster was in the eastern region, from May to October 2021. The first secondary cluster was in the central region, from May to October 2021. The second secondary cluster was in the southeastern region, from May to September 2020. The MaxEnt model showed that the mean annual wind speed, NDVI, cattle density and annual cumulative precipitation were the key factors influencing the occurrence of SFTS. The predicted risk map showed that the area of high prevalence was 28,120 km2, accounting for 18.05% of the total area of the province. CONCLUSIONS: The spatiotemporal distribution of SFTS was heterogeneous and influenced by multidimensional environmental factors. This should be considered as a basis for delineating SFTS risk areas and developing SFTS prevention and control measures.

Genomic diversity and evolution analysis of severe fever with thrombocytopenia syndrome in East Asia from 2010 to 2022.

Background: Conducting an up-to-date analysis on the genomic diversity and evolution patterns of severe fever with thrombocytopenia syndrome virus (SFTSV) is crucial for elucidating the underlying mechanisms of its emergency and pathogenicity, as well as assessing the extent of its threat to public health. Methods: Complete genome sequences of SFTSV were obtained from GenBank until December 19, 2022. A thorough phylogenetic analysis was conducted using comprehensive bioinformatics methods to estimate the genomic diversity and evolution. Results: The phylogenetic classification of SFTSV strains yielded seven lineages (A-G) for each genome segment. SFTSV displayed notable variations in evolutionary patterns among different regions and segments, without a linear accumulation of nucleotide substitutions within segments and regions. The comprehensive analysis revealed 54 recombination events and 17 reassortment strains, including the first discovery of recombination events involving sea-crossing and species-crossing. Selection analysis identified three positive sites (2, 671, 1353) in RNA-dependent RNA polymerase, three positive sites (22, 298, 404) in glycoprotein, and two positive sites (9, 289) in nonstructural protein. No positive selection sites were found in nucleoprotein. Conclusion: Our study unveiled the existence of multiple evolutionary forces influencing SFTSV, contributing to its increasing genetic diversity, which had the potential to modify its antigenicity and pathogenicity. Furthermore, our study highlights the importance of tracking the spread of SFTSV across regions and species.

Soy Protein/Polyvinyl-Alcohol (PVA)-Based Packaging Films Reinforced by Nano-TiO2.

This work was investigated to prepare a reinforcing composite packaging film composited of soy protein/polyvinyl alcohol (PVA) and nano-TiO2. First, different film compositions were designed by the particle size of nano-TiO2, concentration of nano-TiO2, concentration of polyvinylpyrrolidone (PVP, a dispersing agent for nano-TiO2), and pH of film casting solution. Then, the film composition that yielded the optimal physical properties was identified using orthogonal array design single-factor experiments, considering its physical properties, including tensile strength, elongation, water absorption, water vapor transmission, oxygen permeation, thermal property, and film morphology. The results displayed that the optimal film composition was (1) soy protein/PVA film with 2.5 wt% nano-TiO2, (2) 30 nm nano-TiO2 particle size, (3) 1.5 wt% PVP, and (4) pH 6.0 of film-forming solution. It yielded tensile strength of 6.77 MPa, elongation at break rate of 58.91%, and water absorption of 44.89%. Last, the films were characterized by scanning electron microscope (SEM) and differential scanning calorimetry (DSC). SEM analysis showed that compared with the film without TiO2, the film containing TiO2 has a smoother surface, and DSC determined that adding nano-TiO2 can improve the thermostability of soy protein/PVA film. Therefore, the film prepared in this paper is expected to provide a new theoretical basis for use in the packaging industry.

Epidemiological Characteristics of Human Rabies - Shandong Province, China, 2010-2020.

Introduction: Rabies is a viral zoonotic disease that causes progressive and fatal inflammation in the brain. Rabies has caused more than 5,000 human deaths in Shandong Province since 1955. This study aimed to analyze the epidemiological characteristics of human rabies in Shandong Province from 2010 to 2020 and to provide a scientific basis for policy changes. Methods: The data of reported human rabies cases from 2010 to 2020 were obtained from China's National Notifiable Disease Reporting System, and data related to exposure and post-exposure prophylaxis (PEP) of the cases were acquired through case investigation. Results: A total of 414 human rabies cases were reported in Shandong Province from 2010 to 2020. Out of the 414 total cases, 87.20% were primarily farmers; 83.10% were over 40 years old. 70.29% (265/377) belonged to category Ⅲ exposure; and 96.67% (377/390) were exposed to the virus through infected dogs. The vaccine inoculation rate of these cases after exposure was only 8.85%; 1.03% (4/390) had been vaccinated with rabies immunoglobulin, developing the disease 11 to 13 days after category Ⅲ exposure. Conclusions: Dogs were still the primary animal hosts. Most rabies patients died due to no or inadequate post-exposure prophylaxis. Vaccine inoculation rates for dogs should reach the target of 70% as soon as possible. Health departments should improve the accessibility and quality of PEP; and improve the health literacy of the elderly in rural areas.

Survivability of probiotics encapsulated in kelp nanocellulose/alginate microcapsules on microfluidic device.

Probiotics are living microorganisms that can produce health benefits to the host only when they are ingested in sufficient quantities and reach the intestines active state. However, the external environment that probiotics face for a long time before administration and the low pH environment in the stomach after administration can greatly reduce their activity. In this work, we proposed a simple microfluidic encapsulation strategy to efficiently prepare the probiotics-loaded nanocellulose/alginate delivery system, which can improve the storage stability and gastrointestinal survival rate of probiotics. The microcapsules were found to be monodisperse, and the average particle size was<500 µm by observing the microstructure and macroscopic morphology. The kelp nanocellulose was cross-linked in the microcapsule and formed a dense surface with alginate. Through the simulated gastrointestinal digestion experiment, it was found that the survival of probiotics in microcapsules containing 0.5 % and 1.5 % kelp nanocellulose decreased by 1.77 log CFU/g and 1.65 log CFU/g respectively, which was significantly lower than that of nanocellulose-free microcapsules (3.70 log CFU/g). And all the treated groups could release probiotics above 7 log CFU/g after digesting intestinal juice for 6 h. Furthermore, through the storage experiment, it was found that the microcapsules with 1.5 % kelp nanocellulose could still release 8.07 log CFU/g probiotics after four weeks. The results provide a new strategy for probiotics processing and extensive high-value utilization of marine natural products.

Griseocazines: Neuroprotective Multiprenylated Cyclodipeptides Identified through Targeted Genome Mining.

Prenylation can impart pharmacological advantages to bioactive compounds. Global genome mining for prenylated cyclodipeptides identified a gczABC BGC from Streptomyces griseocarneus 132 containing a cyclodipeptide synthase and two prenyltransferase genes. Subsequent heterologous expression allowed isolation and characterization of griseocazines, which displayed potent neuroprotective activity. Further biotransformation analyses revealed that prenyltransferases GczB and GczC catalyzed the stereospecific prenylation of cWW and attached geranyl and farnesyl groups to a cyclodipeptide scaffold, respectively.

Lineage tracing clarifies the cellular origin of tissue-resident macrophages in the developing heart.

Tissue-resident macrophages play essential functions in the maintenance of tissue homeostasis and repair. Recently, the endocardium has been reported as a de novo hemogenic site for the contribution of hematopoietic cells, including cardiac macrophages, during embryogenesis. These observations challenge the current consensus that hematopoiesis originates from the hemogenic endothelium within the yolk sac and dorsal aorta. Whether the developing endocardium has such a hemogenic potential requires further investigation. Here, we generated new genetic tools to trace endocardial cells and reassessed their potential contribution to hematopoietic cells in the developing heart. Fate-mapping analyses revealed that the endocardium contributed minimally to cardiac macrophages and circulating blood cells. Instead, cardiac macrophages were mainly derived from the endothelium during primitive/transient definitive (yolk sac) and definitive (dorsal aorta) hematopoiesis. Our findings refute the concept of endocardial hematopoiesis, suggesting that the developing endocardium gives rise minimally to hematopoietic cells, including cardiac macrophages.

Factors Associated With Severe Fever With Thrombocytopenia Syndrome in Endemic Areas of China.

Objective: To explore the influence of climatic, environmental and socioeconomic factors on SFTS occurrence in Shandong Province, China. Methods: We used generalized additive model to estimate the association between SFTS cases and climatic factors, environmental factors and socioeconomic factors, including annual average temperature, precipitation, land cover, normalized difference vegetation index, altitude, population density, meat production, milk production, and gross domestic product (GDP). Results: There were a total of 4,830 cases reported in 100 (70.9%) counties and districts in Shandong Province from 2010 to 2020. The results showed that the annual average temperature, precipitation, forest and grassland coverage rate, altitude and meat production (square root transform) had a reversed "V" relationship with SFTS occurrence, with the inflection points around 12.5-13.0°C in temperature, around 650 mm in precipitation, around 0.3 in forest and grassland coverage rate, around 300 m in altitude, and around 200-300 tons in meat production (square root transform), respectively. SFTS occurrence had a "V" relationship with milk production (square root transform) and GDP (square root transform), with the inflection points around 100-200 tons in milk production (square root transform), and around 150,000-200,000 yuan in GDP (square root transform), respectively. Conclusions: Climatic, environmental, and socioeconomic factors contributed to the heterogeneous distribution of SFTS in Shandong Province, and the influence of these factors on SFTS occurrence was nonlinear.

Coronary vessel formation in development and regeneration: origins and mechanisms.

Neovascularization of the ischemic myocardium following infarction is vital for the survival of cardiomyocytes and prevention of heart failure. However, the intrinsic revascularization following ischemic injury in the heart is inadequate to restore blood flow to the infarcted myocardium. A comprehensive understanding of how coronary vasculature is constructed and what developmental pathways might be reactivated after infarction is beneficial to develop effective strategies for heart revascularization. The latest lineage tracing studies of coronary vasculature have revealed multiple developmental origins for coronary vascular endothelial cells. The development and growth of coronary vessels emanated from different cellular origins are governed by distinct regulatory mechanisms. Here, we highlight recent research advancements on cellular sources and molecular mechanisms of coronary vessel formation during heart development and regeneration, and also elaborate on how these mechanisms can be reactivated or recapitulated to facilitate therapeutic revascularization in ischemic heart disease.

Extension of Endocardium-Derived Vessels Generate Coronary Arteries in Neonates.

BACKGROUND: Unraveling how new coronary arteries develop may provide critical information for establishing novel therapeutic approaches to treating ischemic cardiac diseases. There are 2 distinct coronary vascular populations derived from different origins in the developing heart. Understanding the formation of coronary arteries may provide insights into new ways of promoting coronary artery formation after myocardial infarction. METHODS: To understand how intramyocardial coronary arteries are generated to connect these 2 coronary vascular populations, we combined genetic lineage tracing, light sheet microscopy, fluorescence micro-optical sectioning tomography, and tissue-specific gene knockout approaches to understand their cellular and molecular mechanisms. RESULTS: We show that a subset of intramyocardial coronary arteries form by angiogenic extension of endocardium-derived vascular tunnels in the neonatal heart. Three-dimensional whole-mount fluorescence imaging showed that these endocardium-derived vascular tunnels or tubes adopt an arterial fate in neonates. Mechanistically, we implicate Mettl3 (methyltransferase-like protein 3) and Notch signaling in regulating endocardium-derived intramyocardial coronary artery formation. Functionally, these intramyocardial arteries persist into adulthood and play a protective role after myocardial infarction. CONCLUSIONS: A subset of intramyocardial coronary arteries form by extension of endocardium-derived vascular tunnels in the neonatal heart.

Strategies for site-specific recombination with high efficiency and precise spatiotemporal resolution.

Site-specific recombinases (SSRs) are invaluable genome engineering tools that have enormously boosted our understanding of gene functions and cell lineage relationships in developmental biology, stem cell biology, regenerative medicine, and multiple diseases. However, the ever-increasing complexity of biomedical research requires the development of novel site-specific genetic recombination technologies that can manipulate genomic DNA with high efficiency and fine spatiotemporal control. Here, we review the latest innovative strategies of the commonly used Cre-loxP recombination system and its combinatorial strategies with other site-specific recombinase systems. We also highlight recent progress with a focus on the new generation of chemical- and light-inducible genetic systems and discuss the merits and limitations of each new and established system. Finally, we provide the future perspectives of combining various recombination systems or improving well-established site-specific genetic tools to achieve more efficient and precise spatiotemporal genetic manipulation.

A suite of new Dre recombinase drivers markedly expands the ability to perform intersectional genetic targeting.

The use of the dual recombinase-mediated intersectional genetic approach involving Cre-loxP and Dre-rox has significantly enhanced the precision of in vivo lineage tracing, as well as gene manipulation. However, this approach is limited by the small number of Dre recombinase driver constructs available. Here, we developed more than 70 new intersectional drivers to better target diverse cell lineages. To highlight their applicability, we used these new tools to study the in vivo adipogenic fate of perivascular progenitors, which revealed that PDGFRa+ but not PDGFRa-PDGFRb+ perivascular cells are the endogenous progenitors of adult adipocytes. In addition to lineage tracing, we used members of this new suite of drivers to more specifically knock out genes in complex tissues, such as white adipocytes and lymphatic vessels, that heretofore cannot be selectively targeted by conventional Cre drivers alone. In summary, these new transgenic tools expand the intersectional genetic approach while enhancing its precision.