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Sirtuins are NAD+-dependent deacetylases with beneficial roles in conditions relevant to human health, including metabolic disease, type II diabetes, obesity, cancer, aging, neurodegenerative diseases, and cardiac ischemia. Since ATP-sensitive K+ (KATP) channels have cardioprotective roles, we investigated whether they are regulated by sirtuins. Nicotinamide mononucleotide (NMN) was used to increase cytosolic NAD+ levels and to activate sirtuins in cell lines, isolated rat and mouse cardiomyocytes or insulin-secreting INS-1 cells. KATP channels were studied with patch clamping, biochemistry techniques, and antibody uptake experiments. NMN led to an increase in intracellular NAD+ levels and an increase in the KATP channel current, without significant changes in the unitary current amplitude or open probability. An increased surface expression was confirmed using surface biotinylation approaches. The rate of KATP channel internalization was diminished by NMN, which may be a partial explanation for the increased surface expression. We show that NMN acts via sirtuins since the increased KATP channel surface expression was prevented by blockers of SIRT1 and SIRT2 (Ex527 and AGK2) and mimicked by SIRT1 activation (SRT1720). The pathophysiological relevance of this finding was studied using a cardioprotection assay with isolated ventricular myocytes, in which NMN protected against simulated ischemia or hypoxia in a KATP channel-dependent manner. Overall, our data draw a link between intracellular NAD+, sirtuin activation, KATP channel surface expression, and cardiac protection against ischemic damage.

Inner coastal wetland ecosystems are generally eutrophic and are often exposed to both salinity stress and Escherichia coli pollution. However, the effects of these stressors on nutrient-cycling and microbial communities are under-researched. Here, we established a vegetated wetland ecosystem in a saline environment to understand the effects of E. coli pollution on nutrient removal and benthic microorganisms. The results show that E. coli significantly inhibited nutrient removal, especially total nitrogen (TN) and ammonium (78.89–84.98 and 3.45–44.65% were removed from the non-E. coli-treated and the E. coli-treated water, respectively). Compared with non-vegetated systems, archaeal community variations at both compositional and phylogenetic levels were weakened in vegetated systems (p < 0.05). Among all the environmental factors, the ratios of PO43--P to total phosphorus and NO3--N to TN contributed the most to archaeal and bacterial community structural variations, respectively. E. coli pollution affected archaeal community succession more than bacteria (p < 0.05). E. coli also weakened the trophic transferring efficiencies between Cyanobacteria and Myxobacteria (p < 0.05). Metabolically, E. coli inhibited bacterial genetic metabolic pathways but made human infection more likely (p < 0.05). Our findings provide new insights into aquatic ecological conservation and environmental management.

The possible mechanism by which the active components of Anhua fuzhuan tea act on FAM in NAFLD lesions was investigated.

The newly described species Gloydius huangi Wang, Ren, Dong, Jiang, Siler et Che, 2019 were described based on only three specimens from two sites. We report a new distribution site from Markam County, Tibet Autonomous Region, China with supplementary description on variation of morphology and mitochondrial genetics of the species. The new specimen varies from types of G. huangi on head scalation, coloration patterns, and hemipenis morphology. A distinct genetical distance 1.9 – 2.2% based on Cytb gene fragment exist between the new specimen and types of the species.

This paper presents three dimensional (3-D) Hall sensors that are capable of detecting magnetic fields in three axis directions and are realized on 180BCDLite® technology with one or two mask adders. The 3-D Hall sensor architecture adopts a modular approach that enables optimization of the planar and vertical Hall devices. The geometries and doping profiles of the Hall devices can be designed for various performance specifications. The planar Hall devices are engineered independently to achieve either a high current–related sensitivity SI >385 V/(A.T) or a high voltage–related sensitivity SV >50 mV/(V.T). For the vertical Hall devices, SV up to 50 mV/(V.T) are demonstrated. The modular design enables flexibility for designers to provide a System-on-Chip (SoC) to meet the needs for a variety of magnetic sensing applications at a low-cost.

Background Increasing evidence indicates that the tumor microenvironment (TME) is a crucial determinant of cancer progression. However, the clinical and pathobiological significance of stromal signatures in the TME, as a complex dynamic entity, is still unclear in esophageal squamous cell carcinoma (ESCC). Methods Herein, we used single-cell transcriptome sequencing data, imaging mass cytometry (IMC) and multiplex immunofluorescence staining to characterize the stromal signatures in ESCC and evaluate their prognostic values in this aggressive disease. An automated quantitative pathology imaging system determined the locations of the lamina propria, stroma, and invasive front. Subsequently, IMC spatial analyses further uncovered spatial interaction and distribution. Additionally, bioinformatics analysis was performed to explore the TME remodeling mechanism in ESCC. To define a new molecular prognostic model, we calculated the risk score of each patient based on their TME signatures and pTNM stages. Results We demonstrate that the presence of fibroblasts at the tumor invasive front was associated with the invasive depth and poor prognosis. Furthermore, the amount of α-smooth muscle actin (α-SMA)+ fibroblasts at the tumor invasive front positively correlated with the number of macrophages (MØs), but negatively correlated with that of tumor-infiltrating granzyme B+ immune cells, and CD4+ and CD8+ T cells. Spatial analyses uncovered a significant spatial interaction between α-SMA+ fibroblasts and CD163+ MØs in the TME, which resulted in spatially exclusive interactions to anti-tumor immune cells. We further validated the laminin and collagen signaling network contributions to TME remodeling. Moreover, compared with pTNM staging, a molecular prognostic model, based on expression of α-SMA+ fibroblasts at the invasive front, and CD163+ MØs, showed higher accuracy in predicting survival or recurrence in ESCC patients. Regression analysis confirmed this model is an independent predictor for survival, which also identifies a high-risk group of ESCC patients that can benefit from adjuvant therapy. Conclusions Our newly defined biomarker signature may serve as a complement for current clinical risk stratification approaches and provide potential therapeutic targets for reversing the fibroblast-mediated immunosuppressive microenvironment.

Mastering the reaction process of underground pyrolysis is the key to oil shale in-situ conversion project. The progress of the pyrolysis reaction can be analyzed by the information of the kerogen maturity in the reaction formation. However, it is difficult to directly test the kerogen maturity by drilling and coring in in-situ conversion project. And the research on judging the process of underground pyrolysis reaction according to the maturity of oil and gas products has not been carried out in depth. This project intends to take the oil shale of the Nenjiang Formation in the southern Songliao Basin and the pyrolysis oil samples produced by the in-situ conversion project as the research object, and conduct simulation experiments and test analysis. The purpose of this paper is to identify the dynamic evolution characteristics of carbon isotope fractionation during the evolution of oil shale pyrolysis hydrocarbon products, and then establish the response relationship with the process of oil shale organic matter pyrolysis. This study has formed a set of dynamic monitoring methods for the in-situ conversion of oil shale to organic matter pyrolysis reaction process, and laid a theoretical foundation for the efficient, economical and stable operation of oil shale in-situ conversion projects.

s Background Giant Cell Arteritis (GCA) is a large vessel vasculitis that most commonly presents with headache, scalp tenderness, jaw claudication, and vision changes. Various other, less common, manifestations have been reported in the literature such as scalp and tongue necrosis. Though most patients respond to corticosteroids, some cases of GCA are refractory to the high doses of corticosteroids. Case presentation We present a 73-year-old female with GCA refractory to corticosteroids presenting with tongue necrosis. This patient significantly improved with a dose of tocilizumab, an IL-6 inhibitor. Conclusion To the best of our knowledge, this is the first case report of a patient with refractory GCA presenting with tongue necrosis that had rapid improvement with tocilizumab. Prompt diagnosis and treatment can prevent severe outcomes such as tongue amputation in GCA patients with tongue necrosis, and tocilizumab may be effective for corticosteroid-refractory cases.

Introduction: Mitochondrial quality control (MQC) is an important mechanism of neural repair after cerebral ischemia (CI). Recent studies have shown that caveolin-1 (Cav-1) is an important signaling molecule in the process of CI injury, but its mechanism of regulating MQC after CI is still unclear. Buyang Huanwu Decoction (BHD) is a classic traditional Chinese medicine formula that is often used to treat CI. Unfortunately, its mechanism of action is still obscure.Methods: In this study, we tested the hypothesis that BHD can regulate MQC through Cav-1 and exert an anti-cerebral ischemia injury effect. We used Cav-1 knockout mice and their homologous wild-type mice, replicated middle cerebral artery occlusion (MCAO) model and BHD intervention. Neurobehavioral scores and pathological detection were used to evaluate neurological function and neuron damage, transmission electron microscopy and enzymology detection of mitochondrial damage. Finally, western blot and RT-qPCR expression of MQC-related molecules were tested.Results: After CI, mice showed neurologic impairment, neuronal damage, and significant destruction of mitochondrial morphology and function, and MQC was imbalanced. Cav-1 deletion aggravated the damage to neurological function, neurons, mitochondrial morphology and mitochondrial function after CI, aggravated the imbalance of mitochondrial dynamics, and inhibited mitophagy and biosynthesis. BHD can maintain MQC homeostasis after CI through Cav-1 and improve CI injury.Discussion: Cav-1 can affect CI injury by regulating MQC, and this mechanism may be another target of BHD for anti-cerebral ischemia injury.

Background: Allergic rhinitis (AR) is strongly associated with a type 2 response, characterized by the cytokines IL-5, IL-4 and IL-13. Several studies have implicated ILC2 and TH2A (CD161+ TH2) but it is not yet entirely clear which subsets are driving the common allergic reactions underlying AR. The objective of this study aims to identify critical pathogenic cell populations associated with AR and to determine their phenotype and functional contribution to disease progression. Methods: We identified integral allergic-specific cell types by transcriptomic sequencing. Whole blood, PBMCs and plasma from a cross-sectional cohort of 216 individuals were analysed by 9-colour flow cytometry and ultra-sensitive cytokine bead arrays using unsupervised clustering algorithms. Clinically active AR cases were further analysed by functional mass cytometry to define phenotype and cytokine secretion (IL-2, IL-3, IL-4, IL-5, IL-9, IL13, IL-17A and IL-22) as well as the expression of the hematopoietic prostaglandin D synthetase (HPGDS). Results: The unbiased analysis revealed that atopy and AR manifestation corelated only with eosinophils, plasma IL-5 and CD161+ TH2 cells. In-depth characterization further revealed that the CD45RB CD161+ TH2 subset were most closely associated with severity. This subset is able to concomitantly secrete multiple cytokines including IL-5, IL-13 and IL-4 and has been previously reported to be associated with other eosinophilic allergies. Conclusion: CD45RB CD161+ TH2 have key roles in driving the allergic response in AR. Neutralizing the CD45RB CD161+ TH2 subset should disrupt the allergic response pathway, thus providing a target for lasting therapeutic interventions.

This study aims to propose and test an integrated model of the theory of planned behaviour (TPB) and uses and gratifications theory (U&G) to understand viewers’ intention to watch video game live streaming on Twitch. Data are collected from 207 Twitch users and analysed using partial least squares structural equation modelling (PLS-SEM) and necessary condition analysis (NCA). Based on the PLS-SEM results, attitudes and subjective norm have a significant positive influence on intention. However, perceived behavioural control does not have a significant influence on intention. Furthermore, entertainment and information seeking have a significant positive influence on attitudes, but socialisation has no significant influence. According to the NCA results, attitudes and perceived behavioural control are the necessary conditions for intention. Moreover, entertainment and information seeking are identified to have necessary effects on attitudes. Subjective norm and socialisation are found to have no necessary effects on intention and attitudes, respectively. This study is the first to investigate the factors that influence viewers’ intention to watch video game live streaming on Twitch based on an integrated perspective of the TPB and U&G theory using the sufficiency (PLS-SEM) and necessary (NCA) logics.

Background Among six extant tiger subspecies, the South China tiger (Panthera tigris amoyensis) once was widely distributed but is now the rarest one and extinct in the wild. All living South China tigers are descendants of only two male and four female wild-caught tigers and they survive solely in zoos after 60 years of effective conservation efforts. Inbreeding depression and hybridization with other tiger subspecies were believed to have occurred within the small, captive South China tiger population. It is therefore urgently needed to examine the genomic landscape of existing genetic variation among the South China tigers. Results In this study, we assembled a high-quality chromosome-level genome using long-read sequences and re-sequenced 29 high-depth genomes of the South China tigers. By combining and comparing our data with the other 40 genomes of six tiger subspecies, we identified two significantly differentiated genomic lineages among the South China tigers, which harbored some rare genetic variants introgressed from other tiger subspecies and thus maintained a moderate genetic diversity. We noticed that the South China tiger had higher FROH values for longer runs of homozygosity (ROH > 1 Mb), an indication of recent inbreeding/founder events. We also observed that the South China tiger had the least frequent homozygous genotypes of both high- and moderate-impact deleterious mutations, and lower mutation loads than both Amur and Sumatran tigers. Altogether, our analyses indicated an effective genetic purging of deleterious mutations in homozygous states from the South China tiger, following its population contraction with a controlled increase in inbreeding based on its pedigree records. Conclusions The identification of two unique founder/genomic lineages coupled with active genetic purging of deleterious mutations in homozygous states and the genomic resources generated in our study pave the way for a genomics-informed conservation, following the real-time monitoring and rational exchange of reproductive South China tigers among zoos.

Supplementary Table S1. Characteristics of the ESCC patients (n=286). Supplementary Table S2. Correlation among clinicopathological parameters and the expression of STAT3β in ESCC patients (n=286).

Supplementary Materials and Methods

Purpose: STAT3 is known to have both oncogenic and tumor suppressive effects, but the regulation of these opposing effects is elusive. We hypothesized that STAT3β, one of the two STAT3 isoforms, is the key determinant in this context.Experimental Design: The prognostic significance of STAT3β and phospho-STAT3αY705 (pSTAT3αY705) was evaluated in 286 cases of patients with esophageal squamous cell carcinoma (ESCC). STAT3β-induced changes in the chemosensitivity to cisplatin and 5-fluorouracil were assessed both in vitro and in vivo. STAT3β-induced changes in the frequency of cancer stem cells were evaluated using Hoechst and CD44 staining. How STAT3β regulates STAT3α was determined using immunoprecipitation, confocal microscopy, DNA-binding, and chromatin immunoprecipitation-PCR.Results: STAT3β expression is an independent protective prognostic marker in patients with ESCC, which strongly correlated with longer overall survival (P = 0.0009) and recurrence-free survival (P = 0.0001). STAT3β significantly decreased the cancer stem cell population, and sensitized ESCC cells to cisplatin and 5-fluorouracil in tumor xenografts. Mechanistically, STAT3β markedly attenuated the transcription activity of STAT3α via inducing STAT3α:STAT3β heterodimers. However, the heterodimer formation decreased the binding between STAT3α and PTPN9 (better known as PTP-MEG2), a protein tyrosine phosphatase, thereby promoting the phosphorylation of STAT3αY705 and enhancing its nuclear translocation and DNA binding. Correlating with this, high STAT3β expression converts the prognostic value of pSTAT3αY705 from unfavorable to favorable in patients with ESCC.Conclusions: STAT3β suppresses chemoresistance and cancer stemness by blocking the transcriptional activity of STAT3α. The paradoxical increase in pSTAT3αY705 induced by STAT3β carries important implications as to how the biologic and prognostic significance of STAT3 in cancers should be interpreted. Clin Cancer Res; 22(3); 691–703. ©2015 AACR.

Supplementary Table S1-S4. Table S1. Correlation between expression of LOXL2 or its spliced isoform L2Δ13 and clinicopathological characteristics in ESCC patients of cohort 1 (n = 252). Table S2. Patient demographics and clinical characteristics for tissue microarray cohort 2 in ESCC (n = 135). Table S3. Multivariate Cox regression for overall survival and disease-free survival of ESCC patients (n = 252). Table S4. Interacting proteins associated with LOXL2 and L2Δ13 in KYSE150 ESCC cells identified by high-performance liquid chromatography followed by tandem mass spectrometry (LC-MS/MS) analysis.

Supplementary Figure Legends

Supplementary Materials and Methods

Supplementary Figure S1. STAT3β suppresses ESCC chemoresistance in vitro. Supplementary Figure S2. STAT3β suppresses cancer stem-like cells in ESCC. Supplementary Figure S3. STAT3β antagonizes the oncogenic function of STAT3α in ESCC cells. Supplementary Figure S4. The complete Western blot data showing the expression of phospho-STAT3α/βTyr705 (pSTAT3α/β) and STAT3α/β in 94 cases of ESCC samples and adjacent non-tumorous tissues. Supplementary Figure S5. STAT3β suppresses STAT3 transcription activity in KYSE150 cells.

Lysyl oxidase-like 2 (LOXL2), a copper-dependent enzyme of the lysyl oxidase family and its nonsecreted, catalytically dead spliced isoform L2Δ13, enhance cell migration and invasion, stimulate filopodia formation, modulate the expression of cytoskeletal genes, and promote tumor development and metastasis in vivo. We previously showed that LOXL2 reorganizes the actin cytoskeleton in esophageal squamous cell carcinoma (ESCC) cells, however, the underlying molecular mechanisms were not identified. Here, using interactome analysis, we identified ezrin (EZR), fascin (FSCN1), heat shock protein beta-1 (HSPB1), and tropomodulin-3 (TMOD3) as actin-binding proteins that associate with cytoplasmic LOXL2, as well as with its L2Δ13 variant. High levels of LOXL2 and L2Δ13 and their cytoskeletal partners correlated with poor clinical outcome in patients with ESCC. To better understand the significance of these interactions, we focused on the interaction of LOXL2 with ezrin. Phosphorylation of ezrin at T567 was greatly reduced following depletion of LOXL2 and was enhanced following LOXL2/L2Δ13 reexpression. Furthermore, LOXL2 depletion inhibited the ability of ezrin to promote tumor progression. These results suggest that LOXL2-induced ezrin phosphorylation, which also requires PKCα, is critical for LOXL2-induced cytoskeletal reorganization that subsequently promotes tumor cell invasion and metastasis in ESCC. In summary, we have characterized a novel molecular mechanism that mediates, in part, the protumorigenic activity of LOXL2. These findings may enable the future development of therapeutic agents targeting cytoplasmic LOXL2.Significance:LOXL2 and its spliced isoform L2Δ13 promote cytoskeletal reorganization and invasion of esophageal cancer cells by interacting with cytoplasmic actin-binding proteins such as ezrin.

Background The existing ex vivo models of endoscopic submucosal dissection (ESD) cannot simulate intraoperative hemorrhage well. We aimed to establish an ESD training method by applying an ex vivo training model with continuous perfusion (ETM-CP). Methods Four training sessions were conducted for 25 novices under the guidance of 2 experts. Eventually, 10 novices completed ESD operations on a total of 89 patients after the training. The resection effectiveness, resection speed, complication rate, and novice performance before and after the training were compared. The data regarding the effects of the training and the model were gathered through a questionnaire survey. Results In terms of the simulation effect of the model, ETM-CP was evaluated as similar to the live pig in all aspects (P > 0.05). The questionnaire analysis revealed that the ESD theoretical knowledge, skill operation, and self-confidence of novices were improved after the training (P P P > 0.05). Conclusion The ETM-CP is effective for ESD training.

Background Carcinosarcoma of the parotid gland is an extremely rare malignancy comprising of 0.04-0.16% of all salivary gland tumors. They consist of distinct carcinomatous and sarcomatous components and may arise de novo or from a preexisting pleomorphic adenoma. Methods and results Herein we present a case of an 80-year-old female who presented with progressively increasing left facial swelling over 6 weeks. Magnetic Resonance Imagining revealed a mass (3.4 cm) in the parotid gland with a predominant cystic/necrotic component. The cytology was atypical (Milan3) and a total parotidectomy and selective lymph node dissection was done. The resection showed extensive necrosis with high grade sarcomatous (chondrosarcoma) areas. The epithelial component was adenoid cystic carcinoma with perineural invasion. The patient is currently undergoing radiotherapy of the tumor bed and skull base due to propensity of perineural invasion of the adenoid cystic component. The most common carcinomas in carcinosarcomas of salivary glands are adenocarcinoma and squamous cell carcinoma. Conclusion This is the first case of an adenoid cystic carcinoma with chondrosarcoma to the best of our knowledge. Carcinosarcoma is a high-grade aggressive lesion with a poor prognosis and should be treated aggressively. More studies are needed to understand the origin of these tumors.

ObjectiveThere are few studies on the prevalence and factors associated with frailty and pre-frailty in older adults with asthma worldwide. The aim of this study was to examine the epidemiological status and factors associated with frailty and pre-frailty in older adults with asthma in China.Research design and methodsData were obtained from the Sample Survey of Aged Population in Urban and Rural China in 2015, a nationwide cross-sectional survey covering 224,142 older people aged 60 years or older in 31 provinces/autonomous regions/municipalities in mainland China. We performed frailty and pre-frailty assessments using the frailty index, and the diagnosis of asthma in the older adults was self-reported based on the history of the physician's diagnosis.ResultsNine thousand four hundred sixteen older adults with asthma were included in the study. The age-sex standardized prevalence of frailty and pre-frailty in Chinese older adults with asthma was 35.8% (95% CI 34.8%−36.7%) and 54.5% (95% CI 53.5%−55.5%). Multinomial logistic regression analysis showed that increased age, female, illiteracy, living alone, poor economic status, ADL disability, comorbid chronic diseases, previous hospitalization in the past year, and residence in northern China were associated with frailty and pre-frailty in older adults with asthma.ConclusionThe prevalence of frailty and pre-frailty in Chinese older adults with asthma is very high, and assessment of frailty should become routine in the management of older adults with asthma. Appropriate public health prevention strategies based on identified risk factors for frailty in older adults with asthma should be developed to reduce the burden of frailty in Chinese older adults with asthma.

The stability of the economic market is an important factor for the rapid development of the economy, especially for the listed companies, whose financial and economic stability affects the stability of the financial market. It is helpful for the healthy development of enterprises and financial markets to make an accurate early warning of the financial economy of listed enterprises. This paper briefly introduced the support vector machine (SVM) and back-propagation neural network (BPNN) algorithms in the machine learning method. To make up for the defects of the two algorithms, they were combined and applied to the enterprise financial economics early warning. A simulation experiment was carried out on the single SVM algorithm-based, single BPNN algorithm-based, and SVM algorithm and BPNN algorithm combined model with the MATLAB software. The results show that the SVM algorithm and BP algorithm combined model converges faster and has higher precision and recall rate and larger area under the curve (AUC) than the single SVM algorithm-based model and the single BPNN algorithm-based model.

To deal with the heat dissipation problem produced by a high integrated circuit, the preparation of heat spreaders with excellent heat transportation performance is increasing in demand. The Ti3C2 MXene sheets and copper particles were fully contacted with cellulose nanofibers by a high-speed mixer, and the composite film was prepared as a heat spreader under the action of the vacuum-assisted filtration. The MXene sheets are connected by the esterification of the carboxyl group in MXene and the hydroxyl group in cellulose nanofibers to form a chemical bond and consist of the main skeleton of the composite film. Due to the synergistic effects of MXene and copper particles, the in-plane and out-of-plane thermal conductivities of the composite film reach 24.96 and 2.46 W m−1 K−1, respectively. Compared with the pure cellulose nanofiber films, the thermal conductivity of composite films increased by 2819.2 and 187.6%, respectively. By designing two applications of composite films in the actual use process, the excellent heating conduction abilities in two directions have been proved. This measure to improve the thermal conductivities of composite films by MXene-copper binary fillers also provides ideas for the novel heat spreader.

ObjectiveTo explore the prevalence and factors associated with frailty and pre-frailty in elderly Chinese patients with hypertension.BackgroundIn China, there have been few national studies into the prevalence and factors associated with frailty and pre-frailty in elderly patients with hypertension.MethodsThrough the 4th Sample Survey of Aged Population in Urban and Rural China (SSAPUR) in 2015, the situation of hypertension subjects aged 60 years or older in 31 provinces, autonomous regions, and municipalities in mainland China was obtained. And the frailty index was constructed based on 33 potential defects, elderly hypertensive patients are classified as robust, frailty, and pre-frailty.ResultsA total of 76,801 elderly patients with hypertension were enrolled in the study. The age-sex standardized prevalence of frailty and pre-frailty in hypertensive elderly in China was 16.1% (95%CI 15.8–16.3%), 58.1% (95%CI 57.7–58.4%). There were significant geographical differences in the prevalence of frailty and pre-frailty in elderly hypertensive patients. Multinomial logistic regression analysis showed that poor economic status, activities of daily living disability, and comorbid chronic diseases were related to frailty and pre-frailty.ConclusionFrailty and pre-frailty are very common in elderly Chinese patients with hypertension and have similar risk factors. Prevention strategies should be developed to stop or delay the onset of frailty by targeting established risk factors in the pre-frailty population of elderly hypertension. It is also crucial to optimize the management of frailty in elderly Chinese patients with hypertension.

Additional file 2: Figure S1. Distribution of 17-mer depth for estimating the genome size. The x-axis represents k-mer depths while y-axis represents the proportions. The blue line represents the proportion of 17-mer in each depth. The peak depth is at 25-fold and total number of 17-mer is 61,791,522,108. The South China tiger (P. t. amoyensis) genome size was estimated to be 2471.66 Mb from the formula: Genome size = K-mer_number/K_depth of peak. Figure S2. Hi-C chromosomal contact heat map. A 500 kb resolution was used to calculate the number of Hi-C read pairs in any two bins. Figure S3. The collinearity between the South China tiger and domestic cat (Felis catus) genomes. Each dot represents an aligned region while the minimum length is 10 kb. The red dot represents forward comparison and the blue dot reverse comparison. Figure S4. Q30 and GC content for each sample. The average Q30 and GC content are 93.24% and 42.04%, respectively. Their very low variation reflected our re-sequencing data to be high quality. Figure S5. The distribution of mapping rates and average mapping depths for each individual. The depths ranged from 12.91× to 18.96 × while the rates varied from 96.03% to 98.92%. Figure S6. The distribution of the SNP number and frequency. Number of genome-wide SNPs (top) and frequency of SNPs per 1 kb of each tiger sample (bottom). Figure S7. Nucleotide diversity π estimates of six tiger subspecies. Figure S8. Heterozygosity statistics of genome-wide SNPs. (a) Observed heterozygosity across all individual genomes of each tiger subspecies. (b) Genomic heterozygosity in each tiger subspecies at population level. Figure S9. Pairwise FST values between six tiger subspecies. The weighted FST values are shown above the diagonal while their standard deviations below the diagonal. Figure S10. Principal component analysis using genome-wide SNPs of six tiger subspecies. Figure S11. A maximum likelihood tree was built using TreeMix software with whole-genome sequencing data of six tiger subspecies and cat. Figure S12. Cluster analysis based on the matrix of genome-wide identity-by-state (IBS) pairwise distances between six tiger subspecies determined by a permutation score. Figure S13. Analysis of genome-wide average identity-by-state (IBS) pairwise identities between six tiger subspecies. Figure S14. Population genetic structure of the South China tigers estimated by the ADMIXTURE. Figure S15. Demographic histories of tiger subspecies. PSMC analysis shows the change in effective population size over time. The dash lines represent 100 bootstraps. The representative individuals sequenced at a high read coverage were selected for each graph, South China tiger (ptam_4, mean 25.38×), Amur tiger (ptal_1, mean 14.76×), Indochinese tiger (ptco_2, mean 13.98×), Malayan tiger (ptja_7, mean 13.13×), Bengal tiger (ptti_3, mean 13.79×), and Sumatran tiger (ptsu_1, mean 13.41×). Figure S16. The results of Dsuite. Heatmap showing statistical support for introgression between pairs of tiger subspecies. Cells in the heatmap indicate the pairwise Z score values between the branch b identified on the expanded tree on the Y axis (relative to its sister branch) and the taxa P3 identified on the X-axis. The grey color indicates the none. (a) Result of branch for South China tiger (lineage 1) with the other tiger subspecies populations. (b) Result of branch for South China tiger (lineage 2) with the other tiger subspecies populations. (c) Result of branch for South China tiger (ptam1 individual) with the other tiger subspecies populations. Figure S17. The graph of ABBA-BABA test. The values of D (pop1, pop2; pop3, pop4) >0 indicates that there are gene flows between pop1 and pop3. Here, only the introgression occurred in South China tiger will be shown. (a) D-statistic for South China tiger (lineage 1) with other tiger subspecies. (b) D-statistic for South China tiger (lineage 2) with other tiger subspecies. (c) D-statistic for South China tiger (ptam1 individual) with other tiger subspecies. Figure S18. Plot of inferred introgression between the South China tiger lineage 1 and other tiger subspecies populations detected by the TreeMix method. The scale bar shows 10 times the average standard error of the entries in the sample covariance matrix. Figure S19. Plot of inferred introgression between the South China tiger lineage 2 and other tiger subspecies populations detected by the TreeMix method. The scale bar shows 10 times the average standard error of the entries in the sample covariance matrix. Figure S20. Plot of inferred introgression between the South China tiger ptam_1 individual and other tiger subspecies populations detected by the TreeMix method. The scale bar shows 10 times the average standard error of the entries in the sample covariance matrix. Figure S21. Genomic inbreeding coefficients FH in each tiger subspecies. Figure S22. The plot of three inbreeding coefficients (FP, FH, and FROH) of the South China tiger. Figure S23. Comparison of homozygote and heterozygote percentage of per mutation category among six tiger subspecies. The significant value was calculated by t-test.

Additional file 1: Fig. S1. Representative images showing the scoring process by the automated quantitative pathology imaging system. Fig. S2. Violin plots displaying the expression level of representative markers in each cell cluster. Fig. S3. Kaplan-Meier survival curves for total α-SMA+ CAFs, lamina propria α-SMA+ CAFs and stromal α-SMA+ CAFs in the generation (n=103) and validation (n=99) dataset of patients with ESCC. Fig. S4. The number of intratumoral macrophages correlates with clinical outcome in ESCC patients. Fig. S5. The density of CD68+ and CD163+ MØs correlates with clinical outcome in patients with ESCC. Fig. S6. Crucial cell-to-cell interaction pathways among the distinct cell populations predicted by CellChat. Fig. S7. Cell-to-cell communication among the CAFs and other cell types. Fig. S8. Differentially-expressed gene (DEG) enrichment analysis for α-SMA+ CAFs. Supplementary Table S1. The clinicopathological parameters of 11 patients profiled by scRNA-seq. Supplementary Table S2. Metal-conjugated antibodies and element-containing reagents used for IMC. Supplementary Table S3. Clinicopathological characteristics in the generation and validation dataset of patients with ESCC. Supplementary Table S4. Correlation between markers and clinicopathological characteristics in the generation and validation datasets. Supplementary Table S5. Differential expressed genes between α-SMA+ CAFs and α-SMA- CAFs. Supplementary Table S6. Univariate and multivariate analyses of factors associated with overall survival (OS) and disease-free survival (DFS) in the generation and validation datasets of patients with ESCC.

PURPOSE We aimed to examine the usefulness of utilizing a specific contrast-enhanced computed tomog raphy (CT) region of interest (ROI) to differentiate renal oncocytoma (RO) from small clear cell renal cell carcinoma (ccRCC) and chromophobe renal cell carcinoma (chRCC). METHODS A retrospective analysis of pre-contrast phase (PCP), corticomedullary phase (CMP), and nephro graphic phase (NP) contrast-enhanced CT images of the histopathologically confirmed initial cohort (27 ROs, 74 ccRCCs, and 36 chRCCs) was conducted. Small, medium, large, and whole ROIs (S-ROI, M-ROI, L-ROI, and W-ROI, respectively) were utilized for CT attenuation value of tumor (AVT), lesion-to-cortex attenuation (L/C), and heterogeneous degree of tumor (HDT) calcula tions. Differences in these parameters were then compared between RO and ccRCC/chRCC, with receiver operating characteristic (ROC) curves being utilized to gauge the diagnostic utility of the statistically significant parameters. Logistic regression analyses were employed to identify key factors capable of differentiating RO and ccRCC/chRCC, with predictive models further being established. A validation cohort (6 ROs, 30 ccRCCs, and 12 chRCCs) was then employed to vali date the performance of the predictive models. RESULTS Of the parameters evaluated using different ROIs, L/C-CMP (S-ROI) (0.88 ± 0.15 vs. 1.13 ± 0.25, P.001) and HDT-CMP (W-ROI) (23.02 (12.00-51.21) vs. 37.81 (16.09-89.45), P.001) were best suited to differentiating RO and ccRCC, yielding respective area under the curve (AUC) values of 0.803 and 0.834. AVT-NP (S-ROI) (122.85 ± 18.87 vs. 86.50 ± 18.65, P.001) and AVT-NP (M-ROI) (119 (86-167) vs. 81.5 (53-142), P.001) were better able to differentiate RO and chRCC, yielding respective AUC values of 0.918 and 0.906. Logistic regression analyses revealed that L/C-CMP (S-ROI) and HDT-PCP, as well as AVT-NP (S-ROI) and HDT-CMP, were the primary factors capable of differentiating RO from ccRCC and chRCC, respectively. The predictive model developed to dif ferentiate between RO and ccRCC exhibited a sensitivity of 66.67% and 55.14% in the initial and validation cohorts, respectively, with corresponding specificity of 94.59% and 93.55%, accuracy of 87.13% and 86.84%, and AUC of 0.908 and 0.876. The predictive model developed to differ entiate between RO and chRCC exhibited a sensitivity of 85.19% and 100.00% in the initial and validation cohorts, respectively, with corresponding specificity of 94.59% and 92.86%, accuracy of 87.30% and 95.24%, and AUC of 0.944 and 0.959. CONCLUSION These data demonstrate that a combination of quantitative parameters measured with particu lar ROIs can enable the efficient and reliable differentiation of RO from ccRCC and chRCC for use in routine patient differential diagnosis.

Fu brick tea (FBT) and its extracts have good lipid-lowering effects and have been used in the treatment of obesity in previous studies. Unfortunately, the therapeutic effect of FBT on non-alcoholic fatty liver disease (NAFLD) has not been thoroughly studied. In this study, we explored the mechanism by which FBT alleviates NAFLD from the perspective of the gut microbiota and liver metabolites. The results showed that FBT could reduce the body weight, liver weight and abdominal fat of NAFLD mice, and improve liver pathological morphology, liver lipid deposition, blood lipids and liver function. Moreover, FBT improved the diversity of the gut microbiota and changed the profile of liver metabolism in NAFLD mice. Further studies showed that FBT could ameliorate the cecum barrier, and regulate the effects of factors related to lipid synthesis in the cecum and liver of NAFLD mice. In conclusion, the present study confirmed that FBT can alleviate high fat induced NAFLD by regulating the homeostasis of the gut microbiota and liver metabolites.

Tea (Camellia sinensis (L.) O. Kuntze), a perennial evergreen shrub, is one of the most important cash crops in China. In September 2021, leaf spot symptoms were observed on approximately 30% of tea plants in a 2 ha commercial field of Lushan (29°33'0" N, 115°58'48" E), Jiangxi Province, China. The symptoms initially appeared as small, gray lesions, and later became larger (10-15 mm in diameter) circular to irregular spots with light brown centers and gray borders. To isolate the pathogen, small pieces (3×3 mm) cut from the margins of lesions were sterilized with 75% ethanol for 10 s, 0.1% HgCl2 for 20 s, and then rinsed three times with sterile water. The pieces were placed onto acidified potato dextrose agar (APDA) plates, and incubated in darkness at 28℃. Pure cultures were prepared by subculturing hyphal tips. A total of 16 fungal isolates were obtained, and the colonies of 15 isolates (isolation rate 93.8%) looked identical, resembling those of the genus Fusarium. The colonies were white to pink with purple woolly mycelium. After 10 to 15 days incubation, slightly curved macroconidia with three to four septa measuring 14.0 to 34.5 × 2.0 to 3.5 µm (n = 50), and oval, unicellar microconidia measuring 4.0 to 9.0 × 1.5 to 3.5 µm (n = 50) were observed. These morphological characteristics were similar to that described for Fusarium proliferatum (Leslie and Summerell 2006). Genomic DNA of representative isolates (LSZWY, LSZWY2, LSZWY3) was extracted with the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech Co., Ltd, Shanghai). The translation elongation factor 1 alpha gene (EF-1ɑ) was amplified using primers EF-1H / EF-2T (O’Donnell, et al. 2015). PCR product was sequenced and the sequence was 709 bp (Accession No. OL614004, ON357634, ON595710). BLAST search results showed that it had 99.9% identity with the EF-1ɑ gene sequence of F. proliferatum (MH341215, MT371378). To test pathogenicity, nine leaves from 5-year-old healthy tea plants (Ca. Luyun 3) were wounded using a sterilized needle and inoculated with a 20µl conidial suspension (2 × 107 conidia·mL-1) on one side of the plants and the other side with sterilized distilled water as a control. All leaves were incubated in a growth chamber at 28℃ and 80% relative humidity with a 12 h light/dark photoperiod. Seven days later, all inoculated treatments showed symptoms identical to those observed in the field, while the control remained asymptomatic. The experiment was repeated three times with similar results. Koch’s postulates were fulfilled by successful re-isolation and morphological and molecular identification of F. proliferatum from the inoculated leaves. This pathogen can cause diseases of many crops, e.g. tobacco, Polygonatum cyrtonema and others (Li, et al 2017; Zhou, et al. 2021). However, this is the first report of F. proliferatum causing leaf spot on tea plants in China. This new disease poses a threat to the yield and quality of tea and methods need to be developed for its control and to prevent further spread.

Size effect has great influence on the shear strength of reinforced concrete T beams, flange width changes the shear mechanism of reinforced concrete T beams. In combination with shear database, comparing the calculation results of formula based on the effective flange width with other specifications formulas by means of Matlab, the flange width can increase the shear capacity of reinforced concrete T beams, and the recommended formula considering the effective flange width is reasonable.

Background Hand, foot and mouth disease (HFMD) is caused by a variety of enterovirus serotypes and the etiological spectrum worldwide has changed since a large scale of outbreaks occurred in 1997. Methods A large number of clinical specimens of HFMD patients were collected in Xiangyang and genotyping was performed by qRT-PCR, conventional PCR amplification and sequencing. Among the 146 CV-A5 detected cases, the complete genome sequences of representative strains were determined for genotyping and for recombination analysis. Results It was found that CV-A5 was one of the six major serotypes that caused the epidemic from October 2016 to December 2017. Phylogenetic analyses based on the VP1 sequences showed that these CV-A5 belonged to the genotype D which dominantly circulated in China. Recombination occurred between the CV-A5 and CV-A2 strains with a breakpoint in the 2A region at the nucleotide 3791. Conclusions The result may explain the emergence of CV-A5 as one of the major pathogens of HFMD. A multivalent vaccine against HFMD is urgently needed to control the disease and to prevent emerging and spreading of new recombinants.

BACKGROUND: We examined the association between the number of resected lymph nodes and survival to determine the optimal lymphadenectomy for thoracic esophageal squamous cell carcinoma (ESCC) patients with negative lymph node. METHODS: We included 1,836 patients from Chinese three high-volumed hospitals with corresponding clinicopathological characters such as gender, age, tumor location, tumor grade and TNM stage of patients. The median follow-up of included patients was 45.7 months (range, 1.03–117.3 months). X-Tile plot was used to identify the lowest number of lymphadenectomy. The multivariate model’s construction was in use of parameters with clinical significance for survival and a nomogram based on clinical variable with P

Per-fluoroalkyl substances (PFASs) have become ubiquitous in farmland ecosystems and pose risks to agricultural safety, and iron is often applied to farmland soils to reduce the availability of pollutants. However, the effects of iron amendment on the availability of PFASs in the soil and on the soil microbiome are not well understood. Here, we investigated the responses of wheat soil containing PFASs to iron addition using a 21-day experiment. Our results showed that iron amendment enhanced PFAS availability (p 0.05) and stimulated superoxide dismutase (SOD) activity in the wheat soil (p 0.05), but iron amendment decreased the activities of soil catalase (CAT) and peroxidase (POD) (p 0.05). Soil bacterial community was more structurally stable than fungal community in response to iron addition, while species' pools were more stable in fungi than in bacteria (p 0.05). Finally, PFPeA's availability in the wheat soil was the most important abiotic factors driving community succession of iron-cycling bacteria (p 0.05). These results highlighted the potential interactions among PFASs' availability and microbial iron cycling in wheat farmland soil ecosystems and provided guidance in farmland environmental conservation and management.

Neurocytomas are rare low-grade brain tumors predominantly affecting young adults, but their cellular origin and molecular pathogenesis is largely unknown. We previously reported a sellar neurocytoma that secreted excess arginine vasopressin causing syndrome of inappropriate anti-diuretic hormone (SIADH). Whole exome sequencing in 21 neurocytoma tumor tissues identified somatic mutations in the plant homeodomain finger protein 14 (PHF14) in 3/21 (14%) tumors. Of these mutations, two were missense mutations and 4 caused splicing site losses, resulting in PHF14 dysfunction. Employing shRNA-mediated knockdown and CRISPR/Cas9-based knockout approaches, we demonstrated that loss of PHF14 increased proliferation and colony formation in five different human, mouse and rat mesenchymal and differentiated cell lines. Additionally, we demonstrated that PHF14 depletion resulted in upregulation of platelet derived growth factor receptor-alpha (PDGFRα) mRNA and protein in neuroblastoma SHSY-5Y cells and led to increased sensitivity to treatment with the PDGFR inhibitor Sunitinib. Furthermore, in a neurocytoma primary culture harboring splicing loss PHF14 mutations, overexpression of wild-type PHF14 and sunitinib treatment inhibited cell proliferation. Nude mice, inoculated with PHF14 knockout SHSY-5Y cells developed earlier and larger tumors than control cell-inoculated mice and Sunitinib administration caused greater tumor suppression in mice harboring PHF-14 knockout than control SHSY-5Y cells. Altogether our studies identified mutations of PHF14 in 14% of neurocytomas, demonstrate it can serve as an alternative pathway for certain cancerous behavior, and suggest a potential role for Sunitinib treatment in some patients with residual/recurrent neurocytoma.