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Abstract Background The development of the human vermiform appendix at the cellular level, as well as its function, is not well understood. Appendicitis in preschool children, although uncommon, is associated with a high perforation rate and increased morbidity. Methods We performed single-cell RNA sequencing (scRNA-seq) on the human appendix during fetal and pediatric stages as well as preschool-age inflammatory appendices. Transcriptional features of each cell compartment were discussed in the developing appendix. Cellular interactions and differentiation trajectories were also investigated. We compared scRNA-seq profiles from preschool appendicitis to those of matched healthy controls to reveal disease-associated changes. Bulk transcriptomic data, immunohistochemistry, and real-time quantitative PCR were used to validate the findings. Results Our analysis identified 76 cell types in total and described the cellular atlas of the developing appendix. We discovered the potential role of the BMP signaling pathway in appendiceal epithelium development and identified HOXC8 and PITX2 as the specific regulons of appendix goblet cells. Higher pericyte coverage, endothelial angiogenesis, and goblet mucus scores together with lower epithelial and endothelial tight junction scores were found in the preschool appendix, which possibly contribute to the clinical features of preschool appendicitis. Preschool appendicitis scRNA-seq profiles revealed that the interleukin-17 signaling pathway may participate in the inflammation process. Conclusions Our study provides new insights into the development of the appendix and deepens the understanding of appendicitis in preschool children. Graphical Abstract

Abstract Background Studies have found that the ratio of total cholesterol to high‐density lipoprotein cholesterol (TC/HDL‐C) was associated with the development of chronic kidney disease (CKD). However, the relationship in different genders was rarely discussed. The aim of this study was to explore this relationship and assess its predictive power for both males and females. Methods Based on a prospective cohort platform in northwest China, 32,351 participants without CKD were collected in the baseline and followed up for approximately 5 years. Cox proportional hazard model and restricted cubic spline regression analysis were performed to investigate the association between TC, HDL‐C, TC/HDL‐C and CKD in adult female and male. The clinical application value of the indicators in predicting CKD was evaluated by the receiver operator characteristic curve. Results During a mean follow‐up of 2.2 years, 484 males and 164 females developed CKD. After adjusted for relevant confounders, for every one standard deviation increase in TC, HDL‐C and TC/HDL‐C, the hazard ratios (HRs) and 95% confidence intervals (95% CIs) for CKD were 1.17 (1.05–1.31), 0.84 (0.71–0.99), and 1.15 (1.06–1.25) for males, 0.94 (0.78–1.13), 0.58 (0.35–0.95), and 1.19 (1.01–1.40) for females, respectively. The results also showed that TC, HDL‐C, and TC/HDL‐C were associated with CKD in a linear dose–response relationship. The TC/HDL‐C had the largest area under the curve (AUC) compared to TC and HDL‐C, and the AUC among the females was larger than that among males. Conclusions The TC/HDL‐C was significantly associated with CKD in adult males and females and has better clinical value in predicting CKD than TC and HDL‐C, especially in females.

Introduction: Torreya grandis is a gymnosperm belonging to Taxodiaceae. As an economically important tree, its kernels are edible and rich in oil with high unsaturated fatty acids, such as sciadonic acid. However, the kernels from different T. grandis landraces exhibit fatty acid and oil content variations. Objectives: As a gymnosperm, does T. grandis have special regulation mechanisms for oil biosynthesis? The aim of this study was to dissect the genetic architecture of fatty acid and oil content and the underlying mechanism in T. grandis. Methods: We constructed a high integrity reference sequence of expressed regions of the genome in T. grandis and performed transcriptome-referenced association study (TRAS) for 10 fatty acid and oil traits of kernels in the 170 diverse T. grandis landraces. To confirm the TRAS result, we performed functional validation and molecular biology experiments for oil significantly associated genes. Results: We identified 41 SNPs from 34 transcripts significantly associated with 7 traits by TRAS (−log10 (P) greater than 6.0). Results showed that LOB domain-containing protein 40 (LBD40) and surfeit locus protein 1 (SURF1) may be indirectly involved in the regulation of oil and sciadonic acid biosynthesis, respectively. Moreover, overexpression of TgLBD40 significantly increased seed oil content. The nonsynonymous variant in the TgLBD40 coding region discovered by TRAS could alter the oil content in plants. Pearson’s correlation analysis and dual-luciferase assay indicated that TgLBD40 positively enhanced oil accumulation by affecting oil biosynthesis pathway genes, such as TgDGAT1. Conclusion: Our study provides new insights into the genetic basis of oil biosynthesis in T. grandis and demonstrates that integrating RNA sequencing and TRAS is a powerful strategy to perform association study independent of a reference genome for dissecting important traits in T. grandis.

Abstract Background Prompt and precise differential diagnosis of biliary atresia (BA) among cholestatic patients is of great importance. Matrix metalloproteinase-7 (MMP-7) holds great promise as a diagnostic marker for BA. This study aimed to investigate the accuracy of age-specific serum MMP-7 for discriminating BA from other cholestatic pediatric patients. Methods This was a single center diagnostic accuracy and validation study including both retrospective and prospective cohorts. Serum MMP-7 concentrations were measured using an ELISA kit, the trajectory of which with age was investigated in a healthy infants cohort aged 0 to 365 days without hepatobiliary diseases (n = 284). Clinical BA diagnosis was based on intraoperative cholangiography and subsequent histological examinations. The diagnostic accuracy of age-specific cutoffs of serum MMP-7 were assessed in a retrospective cohort of cholestatic patients (n = 318, with 172 BA) and validated in a prospective cohort (n = 687, including 395 BA). Results The MMP-7 concentration declines non-linearly with age, showing higher levels in healthy neonates as well as higher cutoff value in neonatal cholestasis. The area under the ROC curve (AUROC) was 0.967 (95% confidence interval [CI]: 0.946–0.988) for the retrospective cohort, and the cutoff of 18 ng/mL yielded 93.0% (95%CI: 88.1-96.3%), 93.8% (95%CI: 88.6-97.1%), 94.7% (95%CI: 90.1-97.5%), and 91.9% (95%CI: 86.4-95.8%) for sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV), respectively. The performance of MMP-7 was successfully validated in the larger prospective cohort, resulting in a diagnostic sensitivity of 95.9% (379/395; 95% CI: 93.5–97.7%), a specificity of 87.3% (255/292; 95% CI: 83.0–90.9%), a PPV of 91.1% (379/416; 95% CI: 87.9–93.7%), and a NPV of 94.1% (255/271; 95% CI: 90.6–96.6%), respectively. Besides, higher cutoff value of 28.1 ng/mL achieved the best sensitivity, specificity, PPV, and NPV for infants aged 0–30 days, which was 86.4% (95% CI: 75.0–94.0%), 95.5% (95% CI: 77.2–99.9%), 98.1% (95% CI: 89.7–100%), and 72.4% (95% CI: 52.8–87.3%), respectively. Conclusions The serum MMP-7 is accurate and reliable in differentiating BA from non-BA cholestasis, showing its potential application in the diagnostic algorithm for BA and significant role in the future research regarding pathogenesis of BA.

ObjectiveTo understand the incidence of carotid atherosclerosis (CAS) and the association of CAS with stroke incidence in an occupational population of the Jinchang Cohort, and to provide an evidence for the prevention and treatment of CAS and stroke. MethodsThe analysis included 6 262 employees of a nonferrous metal company who participated in the Jinchang Cohort Study and had complete information from the baseline examination, including carotid color Doppler ultrasound, from June 2011 to December 2013 and two rounds of follow-up in 2015 and 2017. The cumulative incidence rates of CAS, carotid intima-media thickening, carotid atherosclerotic plaque, and stroke were calculated among the employees during the follow-up period. A multivariate unconditional logistic regression model was used to analyze the factors associated with CAS, carotid intima-media thickening, and carotid atherosclerotic plaque. A multivariate Cox proportional hazards regression model was used to analyze the risk of stroke incidence and its influencing factors in individuals with CAS, carotid intima-media thickening, and carotid atherosclerotic plaque. ResultsDuring the initial follow-up of a mean of 1.71 ± 0.55 years (ranging from 0.08 to 3.75 years), a total of 1 147 cases of CAS, 287 cases of carotid intima-media thickening, and 860 cases of carotid atherosclerotic plaque were observed, with cumulative incidence rates of 18.32%, 4.58%, and 13.73%, respectively. The results of multivariate unconditional logistic regression analysis showed that age ≥ 45 years, smoking, hypertension, diabetes, abnormal total cholesterol (TC), working as a cadre or technical personnel, and internal staff were risk factors for CAS in the occupational population; whereas education level of college or above was a protective factor; age ≥ 45 years, working as a cadre or technical personnel, smoking, having high-sugar diet, and hypertension were risk factors for carotid intima-media thickening, whereas female gender was a protective factor; females, age ≥ 45 years, working as internal staff, hypertension, diabetes, and abnormal TC were risk factors for carotid atherosclerotic plaque, whereas college education or higher was a protective factor. At the second follow-up of a mean of 2.67 ± 0.55 years (ranging from 0.06 to 4.37 years), a total of 240 stroke cases were identified, with a cumulative incidence of 3.83%. The numbers of stroke cases in subjects with CAS, carotid intima-media thickening, and carotid atherosclerotic plaque were 104, 19, and 85, with cumulative stroke incidences of 9.07%, 6.62%, and 9.88%, respectively. The results of the multivariate Cox proportional hazards regression model analysis showed that after adjusting for gender, age, education level, occupation, smoking, alcohol consumption, physical activity, high-salt diet, high-fat diet, high-sugar diet, hypertension, diabetes, hyperuricemia, body mass index (BMI), abnormal TC, abnormal triglycerides (TG), abnormal high-density lipoprotein cholesterol (HDL-C), abnormal low-density lipoprotein cholesterol (LDL-C), and reduced glomerular filtration rate, the risk of stroke was significantly increased in the individuals with CAS (hazard ratio [HR] = 1.421, 95% confidence interval [95%CI]: 1.030 – 1.961) compared with those without CAS. The results also showed that, age ≥ 45 years, high-salt diet, and overweight were risk factors for stroke incidence in the individuals with CAS; females and hyperuricemia were risk factors for stroke incidence in individuals with carotid intima-media thickening; and high school or technical secondary school education, high-salt diet, and overweight were risk factors for stroke incidence in individuals with carotid atherosclerotic plaque. ConclusionThe cumulative incidence of both CAS and stroke was high during follow-up in the occupational population of the Jinchang Cohort Study and in the occupational population, the individuals with CAS had a higher risk of stroke, and older age, high-salt diet, and overweight were the main risk factors for stroke in the individuals with CAS.

Extensive research has established the link between PM2.5 exposure and blood pressure (BP) levels among normal individuals. However, the association between PM2.5 components and BP levels in hypertensive patients has not been fully explored. In this study, 12 971 hypertensive cases from Jinchang cohort (in Jinchang City, China) with nearly 9 years of follow-up were enrolled. Based on the linear mixed-effect model, the effects of fine particulate matter (PM2.5) and five major components [sulfate (SO42-), nitrate (NO3-), ammonium (NH4+), black carbon (BC) and organic matter (OM)]on BP [systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP) and pulse pressure (PP)]were evaluated by single-component model, component-joint model and component-residual model, respectively. A positive correlation was found between PM2.5 as well as its components (SO42-, NO3-, NH4+, BC and OM) exposure and BP levels. The effects of SO42-, BC and OM on BP were observed to be the most robust among the three models. Based on the results of interaction effects and stratified analysis, the effect of BC exposure on SBP, and the effect of PM2.5 and its five components on PP were greater in female than in males. Compared with elderly hypertensive patients, OM had more significant effects on SBP, DBP and MAP in young and (or) middle-aged hypertensive patients. During the heating season, the effect of PM2.5 and its components on BP was grater compared to the non-heating season. Meanwhile, PM2.5 and its components have a greater influence on BP in patients with hypertension combined with diabetes. Therefore, the findings suggested that both PM2.5 exposure and its components had a significant effect on BP in patients with hypertension. Women and young and middle-aged hypertensive patient were the sensitive population. The implementation of source control and reduction of PM2.5 emission (mainly for SO42-, BC and OM) may be of great significance to control BP level and could reduce the risk of cardiovascular disease in patients with hypertension.

Demographic characteristics and clinical data of all newly diagnosed biliary atresia patients in Shanghai were collected from 1 January 2015 to 31 October 2016. The total number of live births was 377 420 during the study period, and the incidence of biliary atresia in Shanghai was 10.86 per 100 000 (95% CI 7.8 to 17.74), with 62.9% and 45.7% cases retaining native liver survival at 2 and 5 years after Kasai procedure, respectively. Implementation of systematic screening measures for biliary atresia in China is needed.

BackgroundLeukemia imposes a large healthcare burden both in China and the United States (US). The disease burden differs greatly between the two countries, but related research is limited. We explored the differences in leukemia incidence and mortality between China and the US.MethodsData on leukemia in China and the US from 1990 to 2021 were collected from the Global Burden of Disease 2021 database. Incidence and mortality were used to estimate the disease burden, and joinpoint regression was performed to compare their secular trends. We used an age-period-cohort model to analyze the effects of age, period, and birth cohort and project future trends in the next 15 years.ResultsIn 2021, the age-standardized incidence rate (ASIR) and the age-standardized death rate (ASDR) of leukemia were lower in China than in the US. However, the incidence and mortality of acute lymphoblastic leukemia (ALL) was considerably higher in China. In the past decades, the ASIR showed decreased tendency in the US, while ASIR showed stable in China. The ASDR tended to decrease in both countries from 1990 to 2021. Males have higher rates of incidence and mortality than females in two countries. The age effects showed that children and older individuals have higher RRs for incidence and mortality in China, while the RRs for incidence and mortality in the US particularly increased in the older population. The disease burden of leukemia in children is obviously greater in China. The ASIRs and ASDRs of leukemia will continue to decline in the next 15 years in China and the US, with the US experiencing a more obvious downtrend.ConclusionsOver the past decades, the ASDRs in two countries both tended to decrease. And compared to the US, China had lower leukemia incidence and mortality, However, the ASIRs in China tended toward stable, which it was showed downtrend in the US. Children have obviously greater RRs for incidence and mortality in China. The incidence and mortality will decrease continuously in two countries. Effective intervention measures are needed to reduce the burden of leukemia.

IntroductionThe expansion of fishery development from offshore to deep-sea areas has become essential for countries to address environmental constraints and ensure food security. This study aims to establish a cooperative development model for deep-blue fishery involving the government, fishery enterprises, and consumers.MethodsWe constructed an evolutionary game and simulation model to analyze the government's deep-blue fishery subsidy strategy. The model considers the interactions between the government, fishery enterprises, and consumers, evaluating how different subsidy strategies influence the development of deep-blue fishery.ResultsOur results demonstrate that government subsidies can stimulate the growth of deep-blue fishery from both the demand and supply sides. Specifically, the likelihood of government subsidies positively affects fishery enterprises' engagement in deep-blue fishery development and consumers' willingness to purchase deep-blue fishery products. The evolutionary game system reaches different steady states based on the cost-benefit changes for the government, fishery enterprises, and consumers, leading to various optimal subsidy strategies.DiscussionThe findings indicate that the amount of government subsidy should be optimized rather than maximized, as excessive subsidies may not yield proportionate benefits. Additionally, subsidizing fishery enterprises proves to be more effective in promoting the development of deep-blue fishery compared to subsidizing consumers. These insights can guide policymakers in designing efficient subsidy strategies to foster sustainable deep-sea fishery development.

Abstract Background Breast cancer is the most common malignant tumor in the world. Intraoperative frozen section of sentinel lymph nodes is an important basis for determining whether axillary lymph node dissection is required for breast cancer surgery. We propose an RRCART model based on a deep-learning network to identify metastases in 2362 frozen sections and count the wrongly identified sections and the associated reasons. The purpose is to summarize the factors that affect the accuracy of the artificial intelligence model and propose corresponding solutions. Methods We took the pathological diagnosis of senior pathologists as the gold standard and identified errors. The pathologists and artificial intelligence engineers jointly read the images and heatmaps to determine the locations of the identified errors on sections, and the pathologists found the reasons (false reasons) for the errors. Through NVivo 12 Plus, qualitative analysis of word frequency analysis and nodal analysis was performed on the error reasons, and the top-down error reason framework of “artificial intelligence RRCART model to identify frozen sections of breast cancer lymph nodes” was constructed based on the importance of false reasons. Results There were 101 incorrectly identified sections in 2362 slides, including 42 false negatives and 59 false positives. Through NVivo 12 Plus software, the error causes were node-coded, and finally, 2 parent nodes (high-frequency error, low-frequency error) and 5 child nodes (section quality, normal lymph node structure, secondary reaction of lymph nodes, micrometastasis, and special growth pattern of tumor) were obtained; among them, the error of highest frequency was that caused by normal lymph node structure, with a total of 45 cases (44.55%), followed by micrometastasis, which occurred in 30 cases (29.70%). Conclusions The causes of identification errors in examination of sentinel lymph node frozen sections by artificial intelligence are, in descending order of influence, normal lymph node structure, micrometastases, section quality, special tumor growth patterns and secondary lymph node reactions. In this study, by constructing an artificial intelligence model to identify the error causes of frozen sections of lymph nodes in breast cancer and by analyzing the model in detail, we found that poor quality of slices was the preproblem of many identification errors, which can lead to other errors, such as unclear recognition of lymph node structure by computer. Therefore, we believe that the process of artificial intelligence pathological diagnosis should be optimized, and the quality control of the pathological sections included in the artificial intelligence reading should be carried out first to exclude the influence of poor section quality on the computer model. For cases of micrometastasis, we suggest that by differentiating slices into high- and low-confidence groups, low-confidence micrometastatic slices can be separated for manual identification. The normal lymph node structure can be improved by adding samples and training the model in a targeted manner.

Abstract Background The efficacy of systemic therapy regimens, such as immune checkpoint inhibitors and tyrosine kinase inhibitors (IO‐TKI) and targeted therapy, for metastatic clear cell renal cell carcinoma (ccRCC) remains unpredictable due to the lack of effective biomarkers. Neutrophil extracellular trap (NET) plays an important role in promoting ccRCC. This study explores the NET predictive value of the efficacy in metastatic ccRCC. Methods In this retrospective study, patients with metastatic ccRCC who received targeted drugs and IO‐TKI were included. Immunofluorescence staining was utilized to quantify the levels of tissue NETs through cell counts of H3Cit(+) and MPO(+) cells. Results A total of 183 patients with metastatic ccRCC were enrolled, including 150 patients who received TKIs and 33 patients who received IO‐TKI. The levels of NETs in tumor tissue were significantly higher than in para‐tumor tissue (p

This study establishes a three-party evolutionary game model that includes marine ranch firms, consumers, and the government, with a focus on the digital transformation pattern of marine ranching. Subsequently, the researchers conduct gaming and simulation to analyze the government subsidy strategy in the digital transformation of marine ranching. The following findings are derived: (1) Government subsidies have the potential to facilitate the digital transformation of marine ranches, benefiting both the supply and demand sides. Government subsidies positively influence both the digital transformation of marine ranch firms and the involvement of consumers in this transformation process. (2) The findings from the evolutionary game system demonstrate that alterations in the cost–benefit dynamics of each participant lead to adjustments in the steady state, thereby prompting the government to adapt its optimal subsidy strategy. (3) More government subsidies are not always better. Excessive government subsidies will destroy the stability of the evolutionary game system, which is not conducive to the realization of the digitization of marine ranch. (4) Subsidizing consumers is more effective in driving the digital transformation of marine ranch than subsidizing marine ranch firms. Subsidies to marine ranch firms will destroy the stability of the evolutionary game system, whereas subsidizing consumers contributes positively to its stability.

The issue of plastic pollution in the marine environment is a matter of great concern. Our research presents pioneering findings on sub-surface microplastics in the Indonesian archipelagic waters. Data on microplastic presence in sub-surface water in the Indonesian Archipelagic Water is crucial for expanding our understanding of microplastic distribution from the surface to the bottom layers of the ocean, a research area that has been relatively overlooked. We discovered microplastic particles at 5 m depth below the surface through simultaneous pump method. The highest concentration of microplastics was discovered in Ambon, followed by North Java and North Sulawesi. In contrast, the lowest abundance of microplastics was detected in the Maluku Sea. These findings are consistent with prior studies establishing a relationship between human activity–as indicated by population density–and microplastic pollution. However, our results indicate that levels of microplastics in Ambon were significantly higher than those found in other sampled areas, particularly compared to North Java, which boasts the highest population density in Indonesia. This disparity is likely due to a faster seawater flushing rate and shorter water residence time in North Java relative to Ambon. The origins and pathways through which these microplastics are introduced into Indonesian archipelagic waters remain uncertain. Based on particle composition (52.73% fibers and 51.38% size

Background: The study of bladder preservation for muscle-invasive bladder cancer (MIBC) mainly focuses on the T2 stage, which remains difficult in the T3 and T4 stage. Pembrolizumab has been applied as neoadjuvant therapy followed by radical cystectomy for MIBC, gaining encouraging results in the phase II study. Disitamab vedotin, an antibody-drug conjugate (ADC), also achieved promising efficacy for refractory bladder cancer. However, the neoadjuvant therapy strategy of these drugs for bladder sparing remains further exploration. Case presentation: A patient with locally advanced MIBC at our institute underwent a neoadjuvant therapeutic regimen followed by transurethral resection of bladder tumor (TURBT) and concurrent chemoradiotherapy. In light of limited initial efficacy, we enacted an adaptive shift in the neoadjuvant treatment strategy, transitioning from a combination of gemcitabine, cis-platinum, and pembrolizumab to disitamab vedotin with pembrolizumab. This approach ultimately achieved bladder preservation, complete response, and a remarkable 1-year disease-free survival (DFS). Conclusion: Proactive evaluation in the early stages of tumor downstaging can serve as a guiding principle for neoadjuvant strategies. This is the first successful case of neoadjuvant pembrolizumab combined with disitamab vedotin and chemotherapy in MIBC patients achieving complete response and bladder preservation.

The implementation of the “Blue Basic Farmland” system for farming ocean is a crucial strategy to ensure food security. This study simulates and analyzes the impact of government fishery subsidies on promoting “Blue Basic Farmland” construction. A tripartite evolutionary game model involving the government, fishermen and fishery enterprises was used”. Subsequently, the fishery subsidy strategy of “Blue Basic Farmland” construction was simulated and analyzed. The following findings are derived. First, fishery subsidies contributed to “Blue Basic Farmland” construction by promoting the participation of fishing enterprises and supporting fishermen. Second, the amounts of fishery subsidies were not the high the better, and different amounts of fishery subsidies had varying effects. Third, subsidizing fishery enterprises was more effective in promoting the construction of “Blue Basic Farmland” compared to subsidizing fishermen. In light of these findings, we proposed the following policy recommendations. The government ought to establish subsidies specifically tailored for “Blue Basic Farmland” construction, while also judiciously controlling the subsidies amounts. Moreover, the focus of these subsidies should be directed towards supporting fishery enterprises.

BackgroundA congenital duodenal diaphragm (CDD) is a rare cause of duodenal obstruction in infants and children. The traditional approach to treatment has been surgical intervention either with excision and duodenoplasty or with bypass through a duodenoduodenostomy, which is invasive and carries the risk of anastomotic leakage.AimsTo summarize the key points of differential diagnosis and treatment of a CDD under gastroscopy and to evaluate its efficacy and safety.MethodsThe clinical data of patients with a duodenal obstruction diagnosed and treated by gastroscopy in our hospital between January 2019 and December 2022 were retrospectively analyzed. The gastroscopic features of the CDD were summarized. The clinical outcomes of patients with CDD treated by gastroscopy were collected and investigated.ResultsA total of 13 children with a duodenal obstruction underwent a gastroscopic examination and/or treatment, and of these, 10 were diagnosed as having a CDD and 3 were finally diagnosed as having an annular pancreas. A dome-shaped structure during inspiration (9/10) and a guidewire that could be inserted through the opening into the distal lumen (6/10) were the gastroscopic features of the CDD, while the annular pancreas had none of these features. The 10 patients, 4 men and 6 women with the CDD, were treated through endoscopic diaphragmotomy and dilation. The age and weight at operation was 15 days to 7 years (M: 2.25 years) and 2.85–22 kg (M: 13.6 kg), respectively. A total of 11 endoscopic operations were performed in the 10 patients. The time of single operation was 15–55 min (M: 38 min). The patients did not experience complications such as bleeding, pneumoperitoneum, and duodenal papilla injury during the operation. Feeding was started 12–24 h after surgery, and the patients were discharged 2–10 days (M: 7 days) postoperatively. The patients were followed up for 3–36 months (M: 17 months), and none of them had a recurrence of vomiting. However, three showed a recurrence of stenosis in upper gastrointestinal imaging, one of whom underwent a partial diaphragm resection again 2 months after the primary treatment.ConclusionsA CDD can be confirmed by endoscopy based on its characteristic features. Endoscopic diaphragmotomy and balloon dilation are safe and effective, which can be a new option for minimally invasive treatment of a CDD.

Objective: This study investigated the predicting value of the ratio of levator hiatus diameter (LHS) to fetal head circumference (HC) in pregnant women at 37 weeks of gestation in the progression of the second stage of labor and levator ani injury 6 weeks postpartum. Methods: A total of 120 first-time women who gave vaginal delivery at 37 weeks of pregnancy were selected as the subjects in our hospital during March 2021 to March 2022. The subjects were divided into the second stage of labor > 1 h group and the second stage of labor ≤ 1 h group, according to the delivery time of the second stage of labor. According to the 6-week postpartum follow-up ultrasound examination with or without levator ani injury, they were divided into levator ani injury group and no injury group. All primipara women underwent three-dimensional ultrasonography at 37 weeks of gestation, and the resting LHS, Valsalva LHS, fetal HC and the ratio of resting LHS and fetal HC were compared. The correlation between these factors and the length of the second labor stage was analyzed using Pearson correlation analysis. The value of these factors in predicting labor progression and postpartum levator ani injury was assessed by receiver operating characteristic (ROC) curves. Results: The resting LHS, Valsalva LHS and the ratio of resting LHS and fetal HC in the levator ani injury group were lower than these in the no injury group, while fetal HC in the levator ani injury group was higher than that in the no injury group (P 1 h group were lower than these in the second stage of labor ≤ 1 h group, while fetal HC in the second stage of labor > 1 h group was higher than that in the second stage of labor ≤ 1 h group (P

Abstract Objective To explore the effect of temperature variability (TV) on admissions and deaths for cardiovascular diseases (CVDs). Method The admissions data of CVDs were collected in 4 general hospitals in Jinchang City, Gansu Province from 2013 to 2016. The monitoring data of death for CVDs from 2013 to 2017 were collected through the Jinchang City Center for Disease Control and Prevention. Distributed lag nonlinear model (DLNM) was combined to analyze the effects of TV (daily temperature variability (DTV) and hourly temperature variability (HTV)) on the admissions and deaths for CVDs after adjusting confounding effects. Stratified analysis was conducted by age and gender. Then the attribution risk of TV was evaluated. Results There was a broadly linear correlation between TV and the admissions and deaths for CVDs, but only the association between TV and outpatient and emergency room (O&ER) visits for CVDs have statistically significant. DTV and HTV have similar lag effect. Every 1 ℃ increase in DTV and HTV was associated with a 3.61% (95% CI: 1.19% ~ 6.08%), 3.03% (95% CI: 0.27% ~ 5.86%) increase in O&ER visits for CVDs, respectively. There were 22.75% and 14.15% O&ER visits for CVDs can attribute to DTV and HTV exposure during 2013–2016. Males and the elderly may be more sensitive to the changes of TV. Greater effect of TV was observed in non-heating season than in heating season. Conclusion TV was an independent risk factor for the increase of O&ER visits for CVDs, suggesting effective guidance such as strengthening the timely prevention for vulnerable groups before or after exposure, which has important implications for risk management of CVDs.

BackgroundPerfluorinated alkyl substances (PFAS) are a class of synthetic organic fluorides, which have adverse health effects on brain function, and limited research has been conducted on their effects on depression. ObjectiveTo assess potential correlation between serum PFAS and depression. MethodsUsing the 2015—2016 and 2017—2018 National Health and Nutrition Examination Survey (NHANES) datasets, 2626 subjects with complete relevant information in people ≥20 years old were selected. Logistic regression and restricted cubic splines were used to analyze the association and dose-response relationship between serum PFAS concentration and depression. Subgroup analysis was performed on sex, age, race, education level, marital status, family income to poverty ratio, moderate exercise, body mass index, and drinking status. ResultsAmong the 2626 subjects, there were 666 patients (25.4%) with mild or above depression. After adjusting for race, education level, marital status, body mass index, moderate exercise, drinking history, cotinine, and other types of PFAS, serum perfluorooctane sulfonate (PFOS) was positively associated with the risk of depression (OR=1.85, 95%CI: 1.14, 3.02), and showed a nonlinear dose-response relationship (χ2=6.37, Pnonlinear=0.012). Perfluorononanoic acid (PFNA) was inversely associated with the risk of depression (OR=0.23, 95%CI: 0.14, 0.39), and showed a linear dose-response relationship (Ptrend

Background: Aidi injection, a classic traditional Chinese medicine (TCM) formula, has been used on a broader scale in treating a variety of cancers. In this study, we aimed to explore the potential anti-tumor effects of Aidi injection in the treatment of neuroblastoma (NB) using network pharmacology (NP).Methods: To elucidate the anti-NB mechanism of Aidi injection, an NP-based approach and molecular docking validation were employed. The compounds and target genes were collected from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMAN-TCM) database. The protein–protein interaction network was constructed using the STRING database. clusterProfiler (R package) was utilized to annotate the bioinformatics of hub target genes. The gene survival analysis was performed on R2, a web-based genomic analysis application. iGEMDOCK was used for molecular docking validation, and GROMACS was utilized to validate molecular docking results. Furthermore, we investigated the anticancer effects of gomisin B and ginsenoside Rh2 on human NB cells using a cell viability assay. The Western blot assay was used to validate the protein levels of target genes in gomisin B- and ginsenoside Rh2-treated NB cells.Results: A total of 2 critical compounds with 16 hub target genes were identified for treating NB. All 16 hub genes could potentially influence the survival of NB patients. The top three genes (EGFR, ESR1, and MAPK1) were considered the central hub genes from the drug–compound–hub target gene–pathway network. The endocrine resistance and estrogen signaling pathways were identified as the therapeutic pathways using the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Gomisin B and ginsenoside Rh2 showed a good binding ability to the target protein in molecular docking. The results of cell experiments showed the anti-NB effect of gomisin B and ginsenoside Rh2. In addition, the administration of gomisin B over-regulated the expression of ESR1 protein in MYCN-amplified NB cells.Conclusion: In the present study, we investigated the potential pharmacological mechanisms of Aidi against NB and revealed the anti-NB effect of gomisin B, providing clinical evidence of Aidi in treating NB and establishing baselines for further research.

ObjectiveTo investigate health-related quality of life (HRQOL) in patients after surgical repair for esophageal atresia (EA) and identify its potential influencing factors.MethodsA total of 102 EA children who had previously visited our hospital participated in this cross-sectional study. Basic data and disease data of the patients were collected. The HRQOL was measured with the Pediatric Quality of Life Inventory™4.0 (PedsQL™4.0) and EA-QOL questionnaire and ranked on a reverse 0–100 scale, with a higher number indicative of a better HRQOL perception. The scores of PedsQL™4.0 in children with EA were collected and compared with that of the demographically matched healthy control group. Meanwhile, the condition-specific HRQOL of EA was analyzed by the EA-QOL questionnaire, and the potential clinical factors that influenced the HRQOL were determined by the generalized linear model.ResultsThe group of EA and control reached a similar score in the generic PedsQL™4.0 (EA group: 86.55 ± 9.69; control group: 89.41 ± 6.54; p = 0.670). There was no significant difference between the EA group and the control group in other domains except the school functioning. Condition-specific HRQOL in the 2–7-year-old group had the highest score in social isolation and stress domain and the lowest score in the physical health and treatment domain, with an overall quality of life score of 83.48 ± 10.22. The scores of the 8–17-year-old group were relatively high in social relationships and health and well-being and lowest in the eating domain, with an overall quality of life score of 89.43 ± 8.57. Heart malformation, complicated esophageal surgery history, respiratory symptoms,and digestive symptoms in the past 1 month were the main factors affecting the HRQOL of children aged 2–7 years. Complicated esophageal surgery history, respiratory symptoms, and digestive symptoms in the past 1 month were the main factors affecting the HRQOL of children aged 8–17 years.ConclusionsThe findings suggest that patients with EA generally had a good HRQOL. However, EA children with postoperative complications and associated symptoms have lower scores in the EA-QOL questionnaire.

PurposeSerum matrix metalloproteinase-7 (MMP-7) is significant in differentiating biliary atresia (BA). This study aims to develop a new peripheral blood quantitative collection device to detect MMP-7 levels via dried blood spot (DBS).MethodsThis is a diagnostic accuracy test. Serum and DBS MMP-7 concentrations were measured using an ELISA kit. Intraoperative cholangiography and subsequent histological examinations were used to confirm BA diagnoses.ResultsA total of 241 infants with obstructive jaundice were enrolled, among whom 168 were BA. Linear regression showed DBS MMP-7 correlated well with serum MMP-7 (R = 0.93, P

The strain sensitivity of bare fiber grating under the action of acoustic wave is too low, which is difficult to detect the effective reflection wavelength change. In this paper, the longitudinal sensing of fiber grating is used as a model, and a layer of thickness far exceeding the inner diameter of the fiber is coated around the grating. Low Young's modulus cylindrical coating is used to increase the strain sensitivity of the fiber grating. We also use the linear region of the reflection spectrum of the phase-shift grating as the edge of the edge filtering method to improve the sensitivity of laser detection. Transmission matrix method is then applied in the simulation to analysis the reflection spectrum of grating and reflectance curve of laser. The simulation results show that the strain sensitivity of the longitudinal sensing model of acoustic wave is better than that of the transverse sensing model. The coating layer with low Young's modulus can significantly improve the strain sensitivity of the grating. Under the action of acoustic wave with frequency lower than 1.7 kHz.The longitudinal sensing sensitivity of the fiber grating coated with polyurethane coating reaches 6.1 nm/MPa, which is 381 times higher than that of the bare fiber grating. Combined with the phase-shift grating reflection spectrum linear region, the comprehensive sensitivity is improved by 2 203.6 times, effectively improving the acoustic wave detection sensitivity of the grating.

Abstract The nuts of Torreya grandis are abundant in flavonoids, providing potential antioxidant activity. Considering that T. grandis can yield nuts for hundreds or thousands of years, effects of tree age on flavonoids and antioxidant activity of T. grandis nuts were targeted in this study for the first time. Results revealed that antioxidant activity and total flavonoid contents in 100‐ and 1000‐years‐old T. grandis nuts were much higher compared to 10‐years‐old T. grandis. The content of 19 flavonoids showed significant correlations with antioxidant activity in 100‐ and 1000‐years‐old trees using metabolites profiling. Transcriptome and coexpression analysis indicated that seven genes were associated with aged‐related flavonoids biosynthesis. Further analysis showed that abscisic acid and gibberellin might regulate the age‐related differential flavonoids accumulation. Our study provides a novel viewpoint that aged T. grandis is likely to present a better source of natural antioxidants and has yet to be examined in other tree models.

Abstract Though diel variations are geographically widespread phenomena among phytoplankton and zooplankton, knowledge is limited regarding diel variations in planktonic ciliate (microzooplankton) community structure. In this study, we analyzed diel variations in community structure of planktonic ciliates in the northern South China Sea (nSCS) and tropical Western Pacific (tWP). Hydrological characteristics during day and night were slightly different over both the nSCS and tWP, while ciliate average abundance at night was clearly higher than in the day in the upper 200 m. In both the nSCS and tWP, abundance proportions of large size-fraction (> 30 μm) aloricate ciliates at night were higher than in the day. While for tintinnids, abundance proportion of large lorica oral diameter at night were lower than in the day. The relationship between environmental factors and ciliate abundance pointed out that depth and temperature were main factors influencing aloricate ciliate and tintinnid in both day and night. For some dominant tintinnid species, chlorophyll a was another important factor influencing their diel vertical distribution. Our results provide fundamental data for better understanding the mechanisms of planktonic ciliate community diel variation in the tropical Western Pacific Ocean.

Torreya grandis is a gymnosperm species that produces edible seeds with high level of sciadonic acid (SCA). Here, the authors assemble the genome of this species, preform methylone analysis of seeds at different developmental stages, and reveal two key genes involved in SCA biosynthesis.

Abstract Background The current study aimed to investigate the dynamic alteration and prognostic significance of tumor‐infiltrating lymphocytes (TILs), tumor‐associated macrophages (TAMs), and PD‐L1 status of immune cells in muscle‐invasive bladder cancer (MIBC) treated with neoadjuvant chemotherapy (NAC). Methods Multiplex immunofluorescence staining was performed to examine CD68+ TAM, CD4+ T cell, CD8+ T cell, FOXP3+ Treg cell, and PD‐L1 expression in paired MIBC tissues (n = 54) before and after NAC. Patients were then divided into definite responders (DR), (≤pT1) and incomplete responders (IR). Results There was no significant difference between DR and IR cohorts for the immune cell infiltration levels at the baseline status. Tobacco history was identified to be associated with worse NAC efficacy. CD68+ (stroma area: p = 0.025; tumor area: p = 0.028; total area: p = 0.013) and CD68+PD‐L1− (stroma area: p = 0.035; tumor area: p = 0.013 total area: p = 0.014) TAMs infiltration levels decreased significantly after NAC, while there was no significant difference of CD68+PD‐L1+ and TILs. The infiltration of CD68+ (p = 0.033), CD68+PD‐L1− (p = 0.033), and CD68+PD‐L1+ (p

Abstract Background Thymic clear cell carcinoma is a rare mediastinal neoplasm, with only 25 reported cases to date. We report a case of a 45-year-old man with thymic clear cell carcinoma. We think imaging and laboratory tests may be helpful for differential diagnosis. Case presentation A 45-year-old male was admitted to a local hospital for chest distress with cardiopalmus. CT showed a mediastinal mass. Laboratory examination results were all in the normal range. Histologically, the tumor cells had a clear cytoplasm, and immunohistochemically, the tumor cells were positive for epithelial markers. We performed abdominal and pelvic CT and further examined serum levels of thyroxine, parathyroid hormone and AFP postoperatively, which were normal. The patient received postoperative radiotherapy, and CT showed left adrenal metastasis at 20 months after surgery. Conclusion Thymic clear cell carcinoma is a rare malignant neoplasm. Adrenal metastasis can occur. Patients undergo thymectomy with chemotherapy or with radiotherapy have better outcoming. Metastasis, direct invasion of parathyroid carcinoma and other primary tumors in the mediastinum should be excluded. Immunohistochemical markers, imaging and laboratory examination can help to exclude metastasis.

Ecological value realization of blue carbon in marine ranching is essential to achieve carbon neutrality. The motivation for conducting the study is to verify whether multi-agent cooperation can promote the ecological value realization of blue carbon in marine ranching. Based on the blue carbon ecological value realization model of marine ranching enterprises, blue carbon demand enterprises, blue carbon trading platforms and government, this paper explores the cooperative governance strategy of marine ranching for each subject using cooperative game and non-cooperative game models. Further, we conduct a comparative analysis to arrive at the optimal strategy. The conclusions are as follows. Multi-agent cooperation is more conducive to the ecological value realization of blue carbon in marine ranching. Compared with non-cooperative governance, the platform's commission and blue carbon price are lower, and the blue carbon output, profit of each market subject, government utility and overall profit are higher in cooperative governance. The strengths of this paper lie in 2 aspects. First, we focuses on the ecological value of blue carbon in marine ranching instead of economic value, providing a new theoretical basis for ecological compensation in marine ranching. Second, we construct a government-led and market-oriented operation of marine ranching's blue carbon ecological value realization mechanism, incorporating blue carbon trading platform and government into the value realization model.

Abstract Neuroblastoma is the most common extracranial solid tumor in children, accounting for 10% to 20% of deaths of pediatric malignancies. Due to the poor prognosis and significant biological heterogeneity of neuroblastoma, it is essential to develop personalized therapeutics and monitor treatment response. Circulating tumor cells (CTCs), as one of the important analytes for liquid biopsy, could facilitate response assessment and outcome prediction for patients in a non‐invasive way. Several methods and platforms have been used for the enrichment and detection of CTCs. The enumeration of CTCs counts and evaluation of tumor‐specific mRNA transcript levels could provide prognostic information at diagnosis, during or after chemotherapy, and during the process of disease progression. So far, studies into neuroblastoma CTCs are only in the preliminary stages. The quality‐controlled large prospective cohort studies are needed to evaluate the clinical significance and statistical rigor of CTC detection methods. Moreover, there remains a lot to be explored and investigated in genotyping characterization of neuroblastoma (NB) CTCs and construction of in‐vitro or in‐vivo functional models. CTCs and circulating tumor DNA (ctDNA) analysis will be complementary in understanding tumor heterogeneity and evolution over the course of therapy for patients with NB in the future.

Abstract The frozen section (FS) diagnoses of pathology experts are used in China to determine whether sentinel lymph nodes of breast cancer have metastasis during operation. Direct implementation of a deep neural network (DNN) in clinical practice may be hindered by misdiagnosis of the algorithm, which affects a patient's treatment decision. In this study, we first obtained the prediction result of the commonly used patch-DNN, then we present a relative risk classification and regression tree (RRCART) to identify the misdiagnosed whole-slide images (WSIs) and recommend them to be reviewed by pathologists. Applying this framework to 2362 WSIs of breast cancer lymph node metastasis, test on frozen section results in the mean area under the curve (AUC) reached 0.9851. However, the mean misdiagnosis rate (0.0248), was significantly higher than the pathologists’ misdiagnosis rate (p 0.01). However, the other low-accuracy group included most of the misdiagnoses of DNN models. Our research shows that the misdiagnosis from deep learning model can be further enriched by our method, and that the low-accuracy WSIs must be selected for pathologists to review and the high-accuracy ones may be ready for pathologists to give diagnostic reports.

BackgroundCurrent evidence on whether occupational sulfur dioxide (SO2) exposure affects the risk of hypertension is still limited, and the research results of the effect of environmental SO2 exposure on risk of hypertension remain inconsistent.ObjectiveTo analyze the association between self-reported occupational exposure to SO2 and the risk of hypertension, and the potential dose-response relationship between the years of exposure to SO2 and the risk of hypertension.MethodsBased on the Jinchang cohort, a nested case-control study design was adopted. A total of 841 newly diagnosed hypertension patients were followed up as the case group, and the control group was selected with 1∶1 individual matching based on non-occupational factors and occupational factors, respectively. The former matching conditions included age ±2 years old, same gender, working age ±2 years, and home address in the same sub-district. The latter was limited to working in the same workshop on the basis of the former conditions. Finally, the former included 717 controls and the latter included 488 controls. A unified questionnaire was used to collect general demographic characteristics, lifestyle habits, history of diabetes, family history of hypertension, and information on occupational exposure to SO2 (self-reported history of occupational exposure to SO2 and years of exposure to SO2). Conditional logistic regression model was used to analyze the association between occupational exposure to SO2 and hypertension, and the dose-response relationship between the years of SO2 exposure and the risk of hypertension.ResultsIn the nested case-control study matching with the non-occupational factors, the OR of hypertension in workers with self-reported occupational exposure to SO2 was 2.39 (95%CI: 1.68-3.39); while when matching with the occupational factors, the OR of hypertension in workers with self-reported occupational exposure to SO2 was 1.48 (95%CI: 1.04-2.12). The results of the dose-response relationship showed that as the SO2 exposure years increased from 1-9 years, 10-19 years, 20-29 years, and 30 years and above, in the nested case-control study matching with non-occupational factors, the ORs of hypertension were 1.85 (95%CI: 0.68-5.08), 1.46 (95%CI: 0.58-3.67), 1.64 (95%CI: 1.00-2.67), and 4.95 (95%CI: 2.63-9.31), respectively; in the nested case-control study matching with occupational factors, the ORs of hypertension were 0.98 (95%CI: 0.40-2.41), 1.84 (95%CI: 0.72-4.70), 1.37 (95%CI: 0.82-2.29), and 2.44 (95%CI: 1.37-4.35), respectively. The two dose-response relationships were positive by χ2 trend test (Ptrend

Mesoscale eddies are energetic and swirling circulations that frequently occur in the open ocean. The effects of mesoscale eddies on the biogeochemical cycling of climate-relevant gases remain poorly constrained. We investigated the distribution and air-sea fluxes of CO2, methane, and five non–methane hydrocarbons (NMHCs) in an anticyclone eddy of Kuroshio Extension during September 2019. Within eddy core, intense stratification hindered the replenishment of nutrients and favored the growth of small-size phytoplankton, such as Prochlorococcus. Seawater pCO2 decreased from 406.1 μatm at the eddy outside to 377.5 μatm at the eddy core, accompanied by a decrease in surface seawater temperature from 26.7 °C to 25.2 °C. The vertical distribution of methane (0.3-9.9 nmol L-1) was influenced by the eddy process, with a maximum at 80 m in the eddy core, which might be attributed to the degradation of phosphonates sustained by Prochlorococcus. The concentrations of five NMHCs (ethane, ethylene, propane, propylene, and isoprene) ranged from 17.2-126.2, 36.7-168.1, 7.5-29.2, 22.6-64.1, 54.5-172.1, 3.5-27.9 pmol L-1, respectively. Isoprene correlated well with Chl-a concentrations at the eddy core, while no significant correlation was observed at the eddy outside. Air-sea fluxes of CO2 and isoprene associated with the eddy core were higher than those of the eddy outside, while the maximum ventilation of methane and other NMHCs (ethane, ethylene, propane, and propylene) was found at the eddy outside. Collectively, physical processes such as eddies impact the production and distribution of light hydrocarbons in seawater and further influence their regional emissions to the atmosphere.

Kaposiform hemangioendothelioma (KHE) is an extremely rare, locally aggressive vascular neoplasm. The etiopathogenesis of KHE is still poorly understood. In the present study, we found a new mutation in KHE (c.685delA, p.Thr229fs). The KHE patient with the PIK3CA mutation showed complete regression after sirolimus treatment. We propose that the presence of the PIK3CA mutation in KHE may correlate with good response to sirolimus.

Torreya grandis is native Chinese tree species of economic significance, renowned for its long lifespan and the rich nutritional value of its nuts. In this study, we analyzed the morphological characteristics, metabolites, associated gene expressions, and regulatory mechanism in nuts from young (10 years old) and old (1000 years old) T. grandis trees. We observed that the length, width, and weight of nuts from older trees were considerably greater than those from younger trees. Metabolomic analysis revealed that the concentrations of 18 amino acids and derivatives (including histidine and serine) in nuts from older trees were markedly higher than those in nuts from younger trees. Transcriptome and metabolomic correlation analysis identified 16 genes, including TgPK (pyruvate kinase), TgGAPDH (glyceraldehyde 3-phosphate dehydrogenase), and others, which exhibit higher expression levels in older trees compared to younger trees, as confirmed by qRT-PCR. These genes are associated with the biosynthesis of histidine, glutamic acid, tryptophan, and serine. Transient expression of TgPK in tobacco led to increased pyruvate kinase activity and amino acid content (histidine, tryptophan, and serine). Additionally, dual-luciferase assays and yeast one-hybrid results demonstrated that TgWRKY21 positively regulates TgPK expression by directly binding to the TgPK promoter. These findings not only demonstrate the nutritional differences between nuts from young and old trees but also offer fresh insights into the development of nutritional sources and functional components based on nuts from old trees, enriching our understanding of the potential benefits of utilizing nuts from older trees.

Background: The pathogenesis of Hirschsprung's disease (HSCR) remains unclear but might involve genes participating in neural crest development. Gene methylation controls the expression of many genes and is involved in the development and migration of neural crest cells, but the involvement of demethylation in HSCR is unknown. This study aimed to investigate the expression of ten-eleven translocation methylcytosine dioxygenase 1 (TET1) (a demethylation protein) in patients with HSCR. Methods: This is a retrospective study of surgical specimens from paediatric patients with and without HSCR (e.g., intussusception and incarcerated hernia) obtained from 07/2015 to 08/2017. TET1 expression was determined by qRT–PCR, western blotting, and immunohistochemistry. The levels of 5-hydroxymethylcytosine were determined by the dot blot assay. Results: The specimens of 35 patients with HSCR and 25 controls were collected. The median TET1 mRNA expression values were 1.028 [HSCR-stenotic (S)], 0.908 [HSCR-dilated (D)], and 0.467 (control) (HSCR-S vs. control: P = 0.002; HSCR-D vs. control: P = 0.008; HSCR-S vs. HSCR-D: P = 0.44). TET1 protein levels followed a similar pattern. The intensity of immunostaining identified higher expression of TET1 in HSCR colon tissues compared with control tissues. The 5-hmC levels in HSCR stenotic segment samples were significantly higher than those in controls. Conclusion: The expression of TET1 is higher in paediatric patients with HSCR than in controls. DNA demethylation initiated by TET1 may be related to HSCR, which demonstrates that TET1 may play a role in the development of HSCR.

Abstract Background Hirschsprung's disease (HSCR) is a relatively common congenital disability. Accumulating extracellular matrix (ECM) prompts intestinal fibrosis remodelling in the aganglionic segments of HSCR. The contributions of various cellular subsets in the fibrogenesis of HSCR segments are poorly understood. Methods Single‐cell transcriptomics from 8 aganglionic segments and 5 normal segments of 7 HSCR subjects and 26 healthy segments of seven healthy donors were analysed. Fibrotic phenotype and alterations were explored using differential expression analysis and single‐cell trajectory analysis. Fibrosis‐related transcription factors were inferred through single‐cell regulatory network inference. Bulk transcriptomic data, proteomic data, immunohistochemistry (IHC) and real‐time polymerase chain reaction were used to validate the alterations in the HSCR intestine. Results Various collagen, fibronectin and laminin protein‐coding genes expression were up‐regulated in the stromal and glial cells of the HSCR intestine. The number of fibroblasts and myofibroblasts in the aganglionic segments increased, and more myofibroblasts were activated at an earlier stage in HSCR segments, which infers that there is an intestinal fibrosis phenotype in HSCR segments. The fibrotic regulators POSTN, ANXA1 and HSP70 were highly expressed in the ECM‐related cellular subsets in the transitional segments and aganglionic segments. The transcription factor regulatory network revealed that fibrosis‐related and megacolon‐related NR2F1 in the fibroblasts and glial subsets was up‐regulated in the aganglionic segment. Conclusions This work identifies intestinal fibrosis and related regulators in aganglionic segments of HSCR; hence, anti‐fibrotic therapy may be considered to prevent HSCR‐associated enterocolitis (HAEC), relieve intestinal stricture and improve cell therapy.

Abstract Background The progression of Biliary Atresia (BA) is associated with the number of reactive ductular cells (RDCs) whose heterogeneity in origin and evolution in humans remains unknown. SOX9-positive liver progenitor-like cells (LPLCs) have been shown to participate in RDCs and new hepatocyte formation during cholestatic liver regeneration in an animal model, which implies the possibility that hepatocyte-reprogrammed LPLCs could be a source of RDCs in BA. The present study aimed to elucidate the characteristics of SOX9-positive LPLCs in BA for exploring new possible therapeutic targets by manipulating the bi-differentiation process of LPLCs to prevent disease progression. Methods Twenty-eight patients, including 24 patients with BA and 4 patients with Congenital Choledochal Cyst as the control group, were retrospectively recruited. Liver biopsy samples were classified histologically using a 4-point scale based on fibrosis severity. LPLCs were detected by SOX9 and HNF4A double positive staining. Single immunohistochemistry, double immunohistochemistry, and multiple immunofluorescence staining were used to determine the different cell types and characteristics of LPLCs. Results The prognostic predictors of BA, namely total bile acid (TBA), RDCs, and fibrosis, were correlated to the emergence of LPLCs. SOX9 and HNF4A double-positive LPLCs co-stained rarely with relevant markers of portal hepatic progenitor cells (portal-HPCs), including CK19, CK7, EPCAM, PROM1 (CD133), TROP2, and AFP. Under cholestasis conditions, LPLCs acquired superior proliferation and anti-senescence ability among hepatocytes. Moreover, LPLCs arranged as a pseudo-rosette structure appeared from the periportal parenchyma to the portal region, which implied the differentiation from hepatocyte-reprogrammed LPLCs to RDCs with the progression of cholestasis. Conclusions LPLCs are associated with disease progression and prognostic factors of BA. The bipotent characteristics of LPLCs are different from those of portal-HPCs. As cholestasis progresses, LPLCs appear to gain superior proliferation and anti-senescence ability and continually differentiate to RDCs.

Whether N6-methyladenosine (m6A) is involved in biliary atresia (BA) remains undefined. Herein, we comprehensively evaluated the m6A profile in BA. When compared with normal controls, BA had an elevated m6A level with upregulated m6A writers. The m6A level was correlated with liver function, stage of fibrosis and jaundice clearance in BA. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) demonstrated an altered m6A topology in BA. MeRIP-seq and RNA sequencing filtered out 130 m6A-modified genes, which were enriched in fibrogenetic pathways. MeRIP-qPCR in vivo and interventions of LX-2 and primary HSCs in vitro validated the regulatory role of m6A on COL1A1 and THY1. THY1+ myofibroblasts expanded in portal area of BA, and highly expressed profibrogenic genes (COL1A1, MMP2, PDGFRA, and DCN). THY1 was correlated with liver fibrosis and jaundice clearance in BA. Bulk array (GSE46960, GSE15235), single-cell RNA sequencing (GSE136103), primary HSC interventions, and co-immunoprecipitation revealed that THY1 was correlated with extracellular matrix organization, promoted HSC activation, showed higher interactions with integrins on myeloid cells in cholestatic fibrosis, and was correlated with native liver survival in BA. Our study highlights the significance of m6A in BA-induced liver fibrogenesis by regulating THY1, shedding new light on the novel therapies to alleviate liver fibrosis by targeting m6A/THY1 axis in BA.

BackgroundSpinal surgery causes severe postoperative pain. An erector spinae plane (ESP) block can relieve postoperative pain, but the optimal blocking method has not been defined. The aim of this study is to compare the feasibility of a one-level and a two-level lumbar ESP block and their effect on intraoperative and postoperative analgesia in lumbar spinal surgery.MethodsA total of 83 adult patients who were scheduled for posterior lumbar interbody fusion were randomly divided into two groups. Patients in Group I (n = 42) received an ultrasound-guided bilateral one-level ESP block with 0.3% ropivacaine, while patients in Group II (n = 41) received a bilateral two-level ESP block. Blocking effectiveness was evaluated, including whether a sensory block covered the surgical incision, sensory decrease in anterior thigh, and quadriceps strength decrease. Intraoperative anesthetic dosage, postoperative visual analogue scale scores of pain, opioid consumption, rescue analgesia, and opioid-related side effects were analyzed.ResultsOf the total number, 80 patients completed the clinical trial and were included in the analysis, with 40 in each group. The time to complete the ESP block was significantly longer in Group II than in Group I (16.0 [14.3, 17.0] min vs. 9.0 [8.3, 9.0] min, P = 0.000). The rate of the sensory block covering the surgical incision at 30 min was significantly higher in Group II than in Group I (100% [40/40] vs. 85.0% [34/40], P = 0.026). The rate of the sensory block in the anterior thigh was higher in Group II (43.8% [35/80] vs. 27.5% [22/80], P = 0.032), but the rate of quadriceps strength decrease did not differ significantly between the groups. The mean effect–site remifentanil concentration during intervertebral decompression was lower in Group II than in Group I (2.9 ± 0.3 ng/ml vs. 3.3 ± 0.5 ng/ml, P = 0.007).There were no significant differences between the groups in terms of intraoperative analgesic consumption, postoperative analgesic consumption, and postoperative VAS pain scores at rest and with movement within 24 h. There were no block failures, block-related complications, and postoperative infection.ConclusionsAmong patients undergoing posterior lumbar interbody fusion, the two-level ESP block provided a higher rate of coverage of the surgical incision by the sensory block when compared with the one-level method, without increasing the incidence of procedure-related complications. Clinical Trial Registrationwww.chictr.org.cn, identifier: ChiCTR2100043596

Currently few studies have explored the relationship between exposure to gaseous pollutants and metabolic health indicators in patients, especially in patients with metabolic syndrome (Mets). This study collected 15,520 patients with Mets in a prospective cohort of nearly 50,000 people with 7 years of follow-up from 2011 to 2017, and matched air pollutants and meteorological data during the same period. The mixed effects model was used to analyze the relationship between different short exposure windows (1-week, 1-month, 2-month, and 3-month) of gaseous pollutants (SO2, NO2, and O3) and the metabolic health indicators of patients after controlled the confounding factors. Stratified analysis was performed by demographic characteristics and behavioral factors. The effects of gaseous pollutants on patients with different Met components were also analyzed. The results showed that the short-term exposure to SO2, NO2, and O3 had a certain effect on the metabolic health indicators of patients with Mets in different exposure windows, and with the extension of the exposure window period, the effects increased. The stratified analysis showed that gender, age, and life behaviors might modify these detrimental effects. In addition, the effects of gaseous pollutants on metabolic health indicators in G4 and G7 were more obvious than other Met components, and the effects of gaseous pollutants on the level of LDL-C were found to be statistically significant in most components. Therefore, patients with Mets should pay more attention to the influence of gaseous pollutants to take appropriate protection to reduce potential health risk.

Background and objectives: Piriform sinus fistula (PSF) is commonly managed by surgery in quiescent inflammation, while medical conditions are hard to be stabilized in patients suffering from recurrent infections. This research attempts to discuss the diagnostic and therapeutic strategies for PSF in acute inflammation. Methods: We retrospectively analyzed the medical records of 113 patients with PSF. Presence of infection within 4 weeks before the operation was taken as a reference to divide the cases into the operation group without infection (Group A) and with infection (Group B). Results: There were 75 cases in Group A and 38 cases in Group B, with an average onset age of 8.16 (5.25-11.58) and 5.63 (4.00-9.54) years, respectively. Esophageal barium swallow identified a positive predictive value of 98.2% in Group A and 100% in Group B. Endoscopic argon gas cauterization was performed in 19 cases (19/75) in Group A, a combination with peripheral doxycycline injection in 42 cases (42/75) and endoscopic- assisted fistulectomy in 14 cases (14/75), compared to 14 cases (14/38), 16 cases (16/38) and 8 cases (8/38) respectively in Group B. The overall recurrence rate was 29.3% in Group A, while those in Group B was 36.8%. Bacterial culture of pus mainly grew Streptococcus in both groups. Conclusion: There was no significant difference in the recurrence rate by EAGC in patients with or without acute infections, and the addition of local doxycycline injection may lead to a decrease of the recurrence rate. Additionally, patients who were complicated with thyroid abscess suffered from a relatively high recurrence rate, requiring specific treatment.

Farmers are the primary decision-makers in chemical fertilizer application. Identifying their chemical fertilizer reduction behavior and its influencing factors is critical to controlling the surface source pollution caused by excessive fertilizer. This paper incorporates farmer characteristic, technology cognition and social capital into the analytical framework of farmers' fertilizer reduction behavior. Based on 889 farmers' questionnaire data, this paper constructs a Structural Equation Model to analyze farmers' fertilizer reduction behavior and its influencing factors from the perspective of farmers' differentiation. The findings are as follows. (1) Farmer characteristics, technology cognition and social capital all positively influences farmers' chemical fertilizer reduction behavior. Among them, social capital has the highest degree of influence on farmers' chemical fertilizer reduction behavior, followed by farmer characteristics, while technology cognition shows the lowest influence. (2) The effect of technology cognition on small-scale pure farmers' chemical fertilizer reduction behavior is insignificant. However, the effect on part-time farmers and large professional farmers is significant, and the effect is higher for large professional farmers. (3) The effect of farmer characteristics on part-time farmers' chemical fertilizer reduction behavior is insignificant. However, the effect on small-scale pure farmers and large professional farmers is significant, and the effect is higher for large professional farmers. (4) The effect of social capital on chemical fertilizer reduction behavior of small-scale pure farmers, part-time farmers, and large professional farmers is significant. This effect is the highest for part-time farmers, followed by large professional farmers, and the lowest for small-scale pure farmers. Accordingly, to continuously promote chemical fertilizer reduction, the government should focus on strengthening policy support, technical support, education guidance and classification.

Abstract Background Biliary atresia (BA) is a devastating inflammatory and fibrosing cholangiopathy of neonates with unknown aetiology. We aim to investigate the relationship between these two main characteristics. Methods Single‐cell RNA sequencing and spatial transcriptomics were performed on liver samples from a cohort of 14 objects (BA: n = 6; control: n = 8). We conducted data integration and cell‐type annotation based on gene expression profiling. Furthermore, we identified fibrosis‐related immune cells according to their spatial locations, GO and KEGG analysis. Finally, SPOTlight and CIBERSORTx were used to deconvolute ST data and microarray data of the GSE46960 cohorts, respectively. Results Immune subpopulations inhabiting the ‘fibrotic niche’ (areas of scarring), comprising ‘intermediate’ CD14++CD16+ monocytes, scar‐associated macrophages, natural killer T cells, transitional B cells and FCN3+ neutrophils were identified. GO and KEGG analyses showed that pathways including ‘positive regulation of smooth muscle cell/fibroblast proliferation’ and ‘positive regulation of/response to VEGFR/VEGF/EGFR/FGF’ were enriched in these cell types. Interactions analysis showed that communication among ‘FGF_FGFR’, ‘RPS19‐C5AR1’, ‘CD74_COPA/MIF/APP’ and ‘TNFRSF1A/B_GRN’ was extensive. Finally, the results of deconvolution for ST data and microarray data validated that the proportions of certain identified fibrosis‐related cell types we identified were increased in BA. Discussion Fibrosis is an important feature of BA, in which the immune system plays an important role. Our work reveals the subpopulations of immune cells enriched in the fibrotic niche of BA liver, as well as key related pathways and molecules; some are highlighted for the first time in liver fibrosis. These newly identified interactions might partly explain why the rate of liver fibrosis occurs much faster in BA than in other liver diseases. Conclusion Our study revealed the molecular, cellular and spatial immune microenvironment of the fibrotic niche of BA.

The recent development of biological sensors has extended marine plankton studies from conducting laboratory bench work to in vivo and real-time observations. Flow cytometry (FCM) has shed new light on marine microorganisms since the 1980s through its single-cell approach and robust detection of the smallest cells. FCM records valuable optical properties of light scattering and fluorescence from cells passing in a single file in front of a narrow-collimated light source, recording tens of thousands of cells within a few minutes. Depending on the instrument settings, the sampling strategy, and the automation level, it resolves the spatial and temporal distribution of microbial marine prokaryotes and eukaryotes. Cells are usually classified and grouped on cytograms by experts and are still lacking standards, reducing data sharing capacities. Therefore, the need to make FCM data sets FAIR (Findability, Accessibility, Interoperability, and Reusability of digital assets) is becoming critical. In this paper, we present a consensus vocabulary for the 13 most common marine microbial groups observed with FCM using blue and red-light excitation. The authors designed a common layout on two-dimensional log-transformed cytograms reinforced by a decision tree that facilitates the characterization of groups. The proposed vocabulary aims at standardising data analysis and definitions, to promote harmonisation and comparison of data between users and instruments. This represents a much-needed step towards FAIRification of flow cytometric data collected in various marine environments.

Jiaozhou Bay is a representative temperate coastal ecosystem and connects with the Yellow Sea by a narrow mouth. The eastern part of the Bay is more polluted than the western and outer parts due to the combined effects of urban sewage and weak water exchange between different water masses. However, the previous results from microscopy analyses did not show any distinct distribution pattern of planktonic ciliates, which are generally considered good bioindicators along environmental gradients. We hypothesize that ciliates may respond to environmental stress by regulating the community of rare taxa, which was not uncovered by morphological methods. The diversity and distribution patterns of planktonic ciliate communities were investigated through high-throughput DNA sequencing from 13 stations in the eastern, western and outer parts of Jiaozhou Bay. The alpha diversity indices showed an increasing trend from the east Bay to the west Bay to the outer Bay. Significant differences in planktonic ciliate communities from different zones of the Bay were mainly attributed to rare rather than abundant taxa. Correlation network analysis revealed that the rare taxa generally have positive correlations with other rare ones, while more negative correlations were found among abundant taxa. We further revealed the distributions of three dominant genera (Stenosemella, Tintinnopsis, and Strombidium) were significantly correlated with environmental variables, suggesting that they might be used as ecological indicators. Our results elucidate the bioindicator potential of ciliate along environmental gradients in a semi-enclosed bay.

Abstract To investigate the mechanism of 25 hydroxyvitamin D (25(OH)D) deficiency in children with biliary atresia (BA) and its effect on liver fibrosis. The serum vitamin D and 25(OH)D, and expression of 25 hydroxylase (CYP2R1 and CYP27A1) in the liver of BA patients were detected and compared with those in the control group. We investigated the effect of differential expression of CYP2R1 in hepatocytes on the expression of genes related to liver fibrosis in primary hepatic stellate cells (HSCs) of BA and animal models of cholestasis. The ratio of 25(OH)D/vitamin D in the BA group was significantly lower than that in the control group. The mRNA and protein expression of CYP2R1 and CYP27A1 in liver tissue of the BA group was significantly lower than that in the control group. Exogenous active vitamin D (calcitriol) inhibited the proliferation and migration of primary HSCs isolated from BA patients, and reduced the expression of fibrosis-related genes in vitro. Downregulation of expression of CYP2R1 in hepatocytes increased expression of transforming growth factor (TGF)-β1, collagen (Col)-1α1 and tissue inhibitor of metalloproteinase (TIMP)-1, and decreased the expression of matrix metalloproteinase (MMP)-2 in cocultured primary HSCs of BA. Upregulation of expression of CYP2R1 in mice with bile duct ligation significantly increased the level of 25(OH)D, decreased the expression of TGF-β1, Col-1α1 and TIMP-1, and increased the expression of MMP-2. Children with BA have impaired vitamin D activation due to CYP2R1 deficiency. The dysactivation of vitamin D can promote the proliferation and activation of HSCs and participate in the development of hepatic fibrosis in BA.

ObjectiveTo investigate the clinical features and prognosis of biliary atresia (BA) with normal or minimally elevated gamma-glutamyl transpeptidase (GGT).MethodsThe clinical data of patients with BA in our hospital between 2012 and 2017 were retrospectively studied. The patients were divided into a low-GGT group (GGT ≤ 300 IU/L) and a high-GGT group (GGT > 300 IU/L) according to the preoperative GGT level. The perioperative clinical parameters, the postoperative jaundice clearance within 6 months, and the 2-year native liver survival were compared among the groups.ResultsA total of 1,998 children were included in this study, namely, 496 in the low-GGT group and 1,502 in the high-GGT group. The ages and weights at the surgery in the low-GGT group were significantly lower than those in the high-GGT group (64.71 ± 21.35 vs. 68.64 ± 22.42 days, P = 0.001; 4.67 ± 1.03 vs. 4.89 ± 0.98 kg, P < 0.001). The levels of serum ALP, ALT, and AST in the low-GGT group were significantly higher than those in the high-GGT group before and 2 weeks after the surgery (ALP: 647.52 ± 244.10 vs. 594.14 ± 228.33 U/L, P < 0.001; ALT: 119.62 ± 97.14 vs. 96.01 ± 66.28 U/L, P < 0.001; AST: 218.00 ± 173.82 vs. 160.71 ± 96.32 U/L; P < 0.001). The INR of the low-GGT group was higher than that of the high-GGT group (1.05 ± 0.34 vs. 0.98 ± 0.20, P < 0.001), while FIB was lower than the high-GGT group (2.54 ± 0.67 vs. 2.73 ± 1.44 g/L; P = 0.006). The decreasing amplitude of TB and DB within 2 weeks after surgery in the low-GGT group was smaller than those in the high-GGT group (TB: 51.62 ± 71.22 vs. 61.67 ± 53.99 μmol/L, P = 0.003; DB: 33.22 ± 35.57 vs. 40.20 ± 35.93 μmol/L, P < 0.001). The jaundice clearance rate in the low-GGT group was significantly lower than that in the high-GGT group at 1, 3, and 6 months after surgery (17.70 vs. 26.05%; 35.17 vs. 48.58%; 38.62 vs. 54.64%, P < 0.001). In addition, the 2-year native liver survival rate in the low-GGT group was significantly lower than that of the high-GGT group (52.5 vs. 66.3%, P < 0.001 HR 1.80, 95% CI 1.38–2.33).ConclusionCompared to patients with high GGT, patients with normal or minimally elevated pre-operative GGT in BA were found to have poorer pre-operative liver function parameters, and post-operatively had lower jaundice clearance rates and worse 2-year native liver survival. This suggests a lower GGT at presentation in biliary atresia could be a sign of more severe liver injury.

The cytokine interferon gamma (IFN-γ) and doxorubicin mono-therapy has been approved by the Food and Drug Administration (FDA) for the treatment of tumors. The importance of IFN-γ in the immune system lies in its immunomodulatory effects, and the importance of doxorubicin in antitumor therapy lies in inhibiting RNA and DNA synthesis. In this work, the role of IFN-γ in the antitumor activity in combination with doxorubicin was investigated. Meanwhile, IFN-γ was used as a vehicle to load doxorubicin over immunotherapy and chemotherapy for synergistic therapy. IFN-γ/doxorubicin complex nanoparticles were prepared by a fusion method with a size of approximately 13 nm and a low polydispersity index. The doxorubicin release profile was analyzed with different pH ranges, and it showed an enhanced release in acidic pH. The ability of IFN-γ/doxorubicin complex nanoparticles to induce human ovarian carcinoma cell (Skov 3) apoptosis was evaluated by the cytotoxicity test. The cellular uptake of IFN-γ/doxorubicin complex nanoparticles was time-dependent, and the IFN-γ/doxorubicin complex nanoparticles showed a higher apoptosis efficiency than free doxorubicin by flow cytometry analysis and fluorescence imaging. This work bridged IFN-γ with doxorubicin to utilize their potential for antitumor activities, opening new avenues for their use in clinical settings.

Sialoblastoma (SBL) is an infrequent embryonal malignant tumor originating from the salivary gland, resembling primitive salivary gland anlage, whereas hepatoblastoma (HB) is the most common pediatric liver malignancy. The simultaneous occurrence of both tumors is extremely rare. Here we reported a case of a 6-month-old infant diagnosed with synchronous SBL and HB. The patient received neoadjuvant chemotherapy followed by surgical resection. Fresh tissues of both tumors were collected before and after chemotherapy, which were further profiled by whole exome sequencing (WES) and single-cell RNA sequencing (scRNA-seq). WES analysis revealed potential somatic driver mutation PIK3CA p.Glu454Lys for SBL and canonical mutation CTNNB1 p.Ser45Pro for HB. No shared somatic variants or common copy number alterations were found between SBL and HB primary tumor samples. Though scRNA-seq, single-cell atlases were constructed for both tumors. SBL may recapitulate a pre-acinar stage in the development of salivary gland, including basaloid, duct-like, myoepithelial-like, and cycling phenotypes. In the meantime, HB was composed of tumor cells resembling different stages of the liver, including hepatocyte-like, hepatic progenitor-like, and hepatoblast-like cells. After chemotherapy, both tumors were induced into a more mature phenotype. In terms of transcriptional signatures, SBL and HB showed enhanced expression of epithelial markers KRT8, KRT18, and essential embryo development genes SDC1, MDK, indicating the disruption of normal embryo epithelium development. Finally, heterozygous deleterious germline mutation BLM and FANCI were identified which could predispose the patient to higher cancer risk. It partially explained the reason for the co-occurrence of SBL and HB. Taken together, we provided valuable resources for deciphering cellular heterogeneity and adaptive change of tumor cells after chemotherapy for synchronous SBL and HB, providing insights into the mechanisms leading to synchronous pediatric tumors.