Delayed gastric emptying after laparoscopic pancreaticoduodenectomy: a single-center experience of 827 cases.

BACKGROUND: Delayed gastric emptying (DGE) commonly occurs after pancreaticoduodenectomy (PD). Risk factors for DGE have been reported in open PD but are rarely reported in laparoscopic PD (LPD). This study was designed to evaluate the perioperative risk factors for DGE and secondary DGE after LPD in a single center. METHODS: This retrospective cohort study included patients who underwent LPD between October 2014 and April 2023. Demographic data, preoperative, intraoperative, and postoperative data were collected. The risk factors for DGE and secondary DGE were analyzed. RESULTS: A total of 827 consecutive patients underwent LPD. One hundred and forty-two patients (17.2%) developed DGE of any type. Sixty-five patients (7.9%) had type A, 62 (7.5%) had type B, and the remaining 15 (1.8%) had type C DGE. Preoperative biliary drainage (p = 0.032), blood loss (p = 0.014), and 90-day any major complication with Dindo-Clavien score ≥ III (p < 0.001) were independent significant risk factors for DGE. Seventy-six (53.5%) patients were diagnosed with primary DGE, whereas 66 (46.5%) patients had DGE secondary to concomitant complications. Higher body mass index, soft pancreatic texture, and perioperative transfusion were independent risk factors for secondary DGE. Hospital stay and drainage tube removal time were significantly longer in the DGE and secondary DGE groups. CONCLUSION: Identifying patients at an increased risk of DGE and secondary DGE can be used to intervene earlier, avoid potential risk factors, and make more informed clinical decisions to shorten the duration of perioperative management.

Automated segmentation and recognition of C. elegans whole-body cells.

MOTIVATION: Motivation: Accurate segmentation and recognition of Caenorhabditis elegans cells are critical for various biological studies, including gene expression, cell lineages and cell fates analysis at single-cell level. However, the highly dense distribution, similar shapes, and inhomogeneous intensity profiles of whole-body cells in 3D fluorescence microscopy images make automatic cell segmentation and recognition a challenging task. Existing methods either rely on additional fiducial markers or only handle a subset of cells. Given the difficulty or expense associated with generating fiducial features in many experimental settings, a marker-free approach capable of reliably segmenting and recognizing C. elegans whole-body cells is highly desirable. RESULTS: We report a new pipeline, called Automated Segmentation and Recognition (ASR) of cells, and applied it to 3D fluorescent microscopy images of L1-stage C. elegans with 558 whole-body cells. A novel displacement vector field based deep learning model is proposed to address the problem of reliable segmentation of highly crowded cells with blurred boundary. We then realize the cell recognition by encoding and exploiting statistical priors on cell positions and structural similarities of neighboring cells. To the best of our knowledge, this is the first method successfully applied to the segmentation and recognition of C. elegans whole-body cells. The ASR segmentation module achieves an F1-score of 0.8956 on a dataset of 116 C. elegans image stacks with 64728 cells (Accuracy 0.9880, AJI 0.7813). Based on the segmentation results, the ASR recognition module achieved an average accuracy of 0.8879. We also show ASR's applicability to other cell types, e.g. platynereis and rat kidney cells. AVAILABILITY: The code is available at https://github.com/reaneyli/ASR. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Effects of denitrification on speciation and redistribution of arsenic in estuarine sediments.

Microbially-mediated redox processes involving arsenic (As) and its host minerals significantly contribute to the mobilization of As in estuarine sediments. Despite its significance, the coupling between As dynamics and denitrification processes in these sediments is not well understood. This study employed sequential sediment extractions and simultaneous monitoring of dissolved iron (Fe), nitrogen (N), and sulfur (S) to investigate the impact of nitrate (NO3-) on the speciation and redistribution of As, alongside changes in microbial community composition. Our results indicated that NO3- additions significantly enhance anaerobic arsenite (As(III)) oxidation, facilitating its immobilization by increased adsorption onto sediment matrices in As-contaminated estuarine settings. Furthermore, NO3- promoted the conversion of As bound to troilite (FeS) and pyrite (FeS2) into forms associated with Fe oxides, challenging the previously assumed stability of FeS/FeS2-bound As in such environments. Continuous NO3- additions ensured As and Fe oxidation, thereby preventing their reductive dissolution and stabilizing the process that reduces As mobility. Changes in the abundance of bacterial communities and correlation analyses revealed that uncultured Anaerolineaceae and Thioalkalispira may be the main genus involved in these transformations. This study underscores the critical role of NO3- availability in modulating the biogeochemical cycle of As in estuarine sediments, offering profound insights for enhancing As immobilization techniques and informing environmental management and remediation strategies in As-contaminated coastal regions.

Independent external validation and comparison of existing pancreatic fistula risk scores after laparoscopic pancreaticoduodenectomy with Bing's pancreaticojejunostomy.

BACKGROUND: The fistula risk score (FRS) is the widely acknowledged prediction model for clinically relevant postoperative pancreatic fistula (CR-POPF). In addition, the alternative FRS (a-FRS) and updated alternative FRS (ua-FRS) have been developed. This study performed external validation and comparison of these 3 models in patients who underwent laparoscopic pancreaticoduodenectomy (LPD) with Bing's pancreaticojejunostomy. METHODS: The FRS total points and predictive probabilities of a-FRS and ua-FRS were retrospectively calculated using patient data from a completed randomized controlled trial. Postoperative pancreatic fistula (POPF) and CR-POPF were defined according to the 2016 International Study Group of Pancreatic Surgery criteria. The correlations of the 4 risk items of the FRS model with CR-POPF and POPF were analyzed and represented using the Cramer V coefficient. The performance of the 3 models was measured using the area under the curve (AUC) and calibration plot and compared using the DeLong test. RESULTS: This study enrolled 200 patients. Pancreatic texture and pathology had discrimination for CR-POPF (Cramer V coefficient: 0.180 vs 0.167, respectively). Pancreatic duct diameter, pancreatic texture, and pathology had discrimination for POPF (Cramer V coefficient: 0.357 vs 0.322 vs 0.257, respectively). Only the calibration of a-FRS predicting CR-POPF was good. The differences among the AUC values of the FRS, a-FRS, and ua-FRS were not statistically significant (CR-POPF: 0.687 vs 0.701 vs 0.710, respectively; POPF: 0.733 vs 0.741 vs 0.750, respectively). After recalibrating, the ua-FRS got sufficient calibration, and the AUC was 0.713 for predicting CR-POPF. CONCLUSION: For LPD cases with Bing's pancreaticojejunostomy, the 3 models predicted POPF with better discrimination than predicting CR-POPF. The recalibrated ua-FRS had sufficient discrimination and calibration for predicting CR-POPF.

A lineage-resolved cartography of microRNA promoter activity in C. elegans empowers multidimensional developmental analysis.

Elucidating the expression of microRNAs in developing single cells is critical for functional discovery. Here, we construct scCAMERA (single-cell cartography of microRNA expression based on reporter assay), utilizing promoter-driven fluorescent reporters in conjunction with imaging and lineage tracing. The cartography delineates the transcriptional activity of 54 conserved microRNAs in lineage-resolved single cells throughout C. elegans embryogenesis. The combinatorial expression of microRNAs partitions cells into fine clusters reflecting their function and anatomy. Notably, the expression of individual microRNAs exhibits high cell specificity and divergence among family members. Guided by cellular expression patterns, we identify developmental functions of specific microRNAs, including miR-1 in pharynx development and physiology, miR-232 in excretory canal morphogenesis by repressing NHR-25/NR5A, and a functional synergy between miR-232 and miR-234 in canal development, demonstrating the broad utility of scCAMERA. Furthermore, integrative analysis reveals that tissue-specific fate determinants activate microRNAs to repress protein production from leaky transcripts associated with alternative, especially neuronal, fates, thereby enhancing the fidelity of developmental fate differentiation. Collectively, our study offers rich opportunities for multidimensional expression-informed analysis of microRNA biology in metazoans.

Phototrophic Nitrogen Fixation, a Neglected Biogeochemical Process in Mine Tailings?

Biological nitrogen fixation (BNF) has important ecological significance in mine tailing by contributing to the initial accumulation of nitrogen. In addition to chemolithotrophic and heterotrophic BNF, light may also fuel BNF in oligotrophic mine tailings. However, knowledge regarding the occurrence and ecological significance of this biogeochemical process in mine tailings remains ambiguous. The current study observed phototrophic BNF in enrichment cultures established from three primary successional stages (i.e., original tailings, biological crusts, and pioneer plants) of tailings. Notably, phototrophic BNF in tailings may be more active at vegetation stages (i.e., biological crusts and pioneering plants) than in bare tailings. DNA-stable isotope probing identified Roseomonas species as potential aerobic anoxygenic phototrophs responsible for phototrophic BNF. Furthermore, metagenomic binning as well as genome mining revealed that Roseomonas spp. contained essential genes involved in nitrogen fixation, anoxygenic photosynthesis, and carbon fixation, suggesting their genetic potential to mediate phototrophic BNF. A causal inference framework equipped with the structural causal model suggested that the enrichment of putative phototrophic diazotrophic Roseomonas may contribute to an elevated total nitrogen content during primary succession in these mine tailings. Collectively, our findings suggest that phototrophic diazotrophs may play important roles in nutrient accumulation and hold the potential to facilitate ecological succession in tailings.

Bacterial community in the metal(loid)-contaminated marine vertical sediments of Jinzhou Bay: Impacts and adaptations.

Metal(loid) discharge has led to severe coastal contamination; however, there remains a significant knowledge gap regarding its impact on sediment profiles and depth-resolved bacterial communities. In this study, geochemical measurements (pH, nutrient elements, total and bioavailable metal(loid) content) consistently revealed decreasing nitrogen, phosphorus, and metal(loid) levels with sediment depth, accompanied by reduced alpha diversity. Principal coordinate analysis indicated distinct community compositions with varying sediment depths, suggesting a geochemical influence on diversity. Ecological niche width expanded with depth, favoring specialists over generalists, but both groups decreased in abundance. Taxonomic shifts emerged, particularly in phyla and families, correlated with sediment depth. Microbe-microbe interactions displayed intricate dynamics, with keystone taxa varying by sediment layer. Zinc and arsenic emerged as key factors impacting community diversity and composition using random forest, network analysis, and Mantel tests. Functional predictions revealed shifts in potential phenotypes related to mobile elements, biofilm formation, pathogenicity, N/P/S cycles, and metal(loid) resistance along sediment profiles. Neutral and null models demonstrated a transition from deterministic to stochastic processes with sediment layers. This study provides insights into the interplay between sediment geochemistry and bacterial communities across sediment depths, illuminating the factors shaping these ecosystems.

A full-body transcription factor expression atlas with completely resolved cell identities in C. elegans.

Invariant cell lineage in C. elegans enables spatiotemporal resolution of transcriptional regulatory mechanisms controlling the fate of each cell. Here, we develop RAPCAT (Robust-point-matching- And Piecewise-affine-based Cell Annotation Tool) to automate cell identity assignment in three-dimensional image stacks of L1 larvae and profile reporter expression of 620 transcription factors in every cell. Transcription factor profile-based clustering analysis defines 80 cell types distinct from conventional phenotypic cell types and identifies three general phenotypic modalities related to these classifications. First, transcription factors are broadly downregulated in quiescent stage Hermaphrodite Specific Neurons, suggesting stage- and cell type-specific variation in transcriptome size. Second, transcription factor expression is more closely associated with morphology than other phenotypic modalities in different pre- and post-differentiation developmental stages. Finally, embryonic cell lineages can be associated with specific transcription factor expression patterns and functions that persist throughout postembryonic life. This study presents a comprehensive transcription factor atlas for investigation of intra-cell type heterogeneity.

Extended pancreatic neck transection versus conventional pancreatic neck transection during laparoscopic pancreaticoduodenectomy (LPDEXCEPT): protocol for a multicentre superiority randomised controlled trial.

INTRODUCTION: Postoperative pancreatic fistula (POPF) remains one of the most severe complications of laparoscopic pancreaticoduodenectomy (LPD). Theoretically, transecting the pancreatic neck more distally has both advantages (more blood supply, and more central pancreatic duct) and disadvantages (maybe smaller the pancreatic duct) in preventing POPF. This theoretical contradiction pushed us to organise this trial to explore the impact of the level of pancreatic transection in clinical practice. We conduct this randomised trial with the hypothesis that extended pancreatic neck transection has superiority to conventional pancreatic neck transection. METHODS AND ANALYSIS: The LPDEXCEPT (Extended pancreatic neck transection versus conventional pancreatic neck transection during laparoscopic pancreaticoduodenectomy) trial is a multicentre, randomised-controlled, open-label, superiority trial in 4 centres whose annual surgical volume for LPD is more than 25 cases with pancreatic surgeons who had completed their learning curve. A total of 154 patients who meet the inclusive and exclusive criteria are randomly allocated to the extended pancreatic neck transection group or conventional pancreatic neck transection group in a 1:1 ratio. The stratified randomised block design will be applied, with stratified factors are surgical centre and the diameter of the main pancreatic duct measured by preoperative CT scan (preMPD). The primary outcome is the incidence of the clinically relevant pancreatic fistula. ETHICS AND DISSEMINATION: Ethics Committee on Biomedical Research of West China Hospital of Sichuan University has approved this trial in March 2023 (approval no. 2023-167). Results of this trial will be published in peer-reviewed journals and conference proceedings. TRIAL REGISTRATION NUMBER: NCT05808894.

Divergent adaptation strategies of abundant and rare bacteria to salinity stress and metal stress in polluted Jinzhou Bay.

Understanding how abundant (AT) and rare (RT) taxa adapt to diverse environmental stresses is vital for assessing ecological processes, yet remains understudied. We collected sediment samples from Liaoning Province, China, representing rivers (upstream of wastewater outlet), estuaries (wastewater outlets), and Jinzhou Bay (downstream of wastewater outlets), to comprehensively evaluate AT and RT adaptation strategies to both natural stressors (salinity stress) and anthropogenic stressors (metal stress). Generally, RT displayed higher α- and ß-diversities and taxonomic groups compared to AT. Metal and salinity stresses induced distinct α-diversity responses in AT and RT, while ß-diversity remained consistent. Both subcommunities were dominated by Woeseia genus. Metal stress emerged as the primary driver of diversity and compositional discrepancies in AT and RT. Notably, AT responded more sensitively to salinity stress than RT. Stress increased topological parameters in the biotic network of AT subcommunities while decreasing values in RT subcommunities, concurrently loosening interactions of AT with other taxa and strengthening interactions of RT with others in biotic networks. RT generally exhibited greater diversity of metal resistance genes compared to AT. Greater numbers of genes related to salinity tolerance was observed for the RT than for AT. Compared to AT, RT demonstrated higher diversity of metal resistance genes and a greater abundance of genes associated with salinity tolerance. Additionally, deterministic processes governed AT community assembly, reinforced by salinity stress. However, the opposite trend was observed in the RT, where the importance of stochastic process gradually increased with metal stresses. The study is centered on exploring the adaptation strategies of both AT and RT to environmental stress. It underscores the importance of future research incorporating diverse ecosystems and a range of environmental stressors to draw broader and more reliable conclusions. This comprehensive approach is essential for gaining a thorough understanding of the adaptive mechanisms employed by these microorganisms.

Predictive Value of a Novel Baseline Diffusion-Weighted Imaging Posterior Circulation Score in Endovascular Treatment of Patients with Acute Vertebrobasilar Occlusion.

RATIONALE AND OBJECTIVES: To investigate the predictive value of a novel posterior circulation score (novel-PC score) based on baseline posterior circulation diffusion-weighted imaging (DWI) for functional independence after endovascular treatment (EVT) in patients with acute vertebral-basilar artery occlusion (VBAO). MATERIALS AND METHODS: The baseline DWI brain stem score (BSS), posterior circulation Alberta Stroke Program Early CT Score (pc-ASPECTS), and the novel-PC score were evaluated separately. A modified Rankin scale (mRS) ≤2 at 90 days was defined as a prognostic indicator of functional independence. Modified Thrombolysis in Cerebral Infarction grade 2b or 3 was defined as surgical success. RESULTS: A total of 64 eligible patients were enrolled and divided into good and poor functional prognosis groups based on the mRS. The novel-PC score, BSS, and pc-ASPECTS (all P ≤ .001) were significantly better in the good functional prognosis group. The novel-PC score had a higher predictive value than BSS and pc-ASPECTS for 90-day functional independence (area under the receiver operating characteristic curve, 0.87 vs. 0.73 vs. 0.71; P < .05). Univariate binary logistic regression analysis showed that age (P = .006), Posterior National Institutes of Health Stroke Scale ≤18 (P < .001), BSS ≤2 (P = .008), pc-ASPECTS >7 (P = .002), and novel-PC score ≤5 (P = .001) were independently associated with function. CONCLUSION: Our novel-PC score, based on DWI, can independently predict functional prognosis in patients with acute VBAO after EVT. CLINICAL RELEVANCE: The novel-PC score based on baseline DWI was shown to be an independent predictor of function in patients with acute BVAO who are treated with EVT.

General cognitive processing for orthographic discrepancy engages foveal attention during sentence comprehension.

Words are processed in the parafovea and fovea in succession during natural reading, but the classic rapid serial visual presentation (RSVP) paradigm presents words only in the fovea. Unlike the RSVP paradigm, the RSVP with flanker (RSVP-flanker) paradigm is similar to natural text reading. Previous studies using the RSVP-flanker paradigm have suggested that high-level semantic/syntactic integration engages foveal fixation after parafoveal semantic access. However, it is less clear how general cognitive processing, such as discrepancy monitoring and error correction, unfolds across the parafoveal and foveal visual fields. In the current study, Chinese sentences were presented with the RSVP-flanker paradigm. Critical words were manipulated so that they were expected (EXP), semantically violated (VIO), or orthographically similar (ORT). Because of the nuanced differences between the EXP and ORT conditions, the processing of ORT critical words required more general cognitive processing, such as discrepancy monitoring and error correction. In contrast, the processing of VIO words may require more semantic integration. The ERP results showed more positive foveal late positive component (LPC) in ORT versus VIO. Moreover, there was no parafoveal LPC effect, but a robust foveal LPC effect, in the contrasts of VIO/ORT versus EXP, replicating previous results. Together, the results provide substantial evidence that general cognitive processing for orthographic discrepancy occurs at the foveal perception.

Cadmium-chelating ability of the siderophore DHBS secreted by Leclercia adecarboxylata FCH-CR2 and its action mechanism.

As one of the most accumulative toxic heavy metals, cadmium (Cd) poses a major threat to human health. Bacterial siderophores, as small molecules with metal-absorbing ability, have great potential activity for Cd-reduction. In this study, the siderophore-producing bacterialstrain FCH-CR2 was isolated from a high-Cd contaminated soil using the CAS method. Leclercia adecarboxylata was identified through 16S rRNA sequence, homology analysis, colony morphology, physiological and biochemical tests. A siderophore, catechol type 2,3-dihydroxy-N-benzoyl-l-serine (DHBS) secreted by FCH-CR2, was purified using RP-HPLC and identified by LC-MS/MS. Intraperitoneal injection of DHBS significantly increased fecal Cd levels, and reduced Cd accumulation in organs. In density flooding theory (DFT) analysis, DHBS may bind to Cd via the hydroxyl site on the benzene ring. Besides, the isothermal titration calorimetry (ITC) assay revealed that the formation of Cd-DHBS is a spontaneous and endothermic reaction with ΔG = -21.4 kJ/mol and ΔH = 1.51 ± 0.142 kJ/mol.

Multiplex PCR-based next generation sequencing as a novel, targeted and accurate molecular approach for periprosthetic joint infection diagnosis.

Objectives: Periprosthetic joint infection (PJI) diagnosis remains challenging, and the identification of the causative microorganism is, by far, the most important aspect. Here, we use multiple PCR-based targeted next-generation sequencing (tNGS) to detect pathogens in PJI. To explore 1. the ability of targeted next-generation sequencing (tNGS) to detect pathogens in PJI; 2. the consistency of tNGS, metagenomic NGS (mNGS), and culture results; and 3. the ability of tNGS to detect drug resistance genes in PJI. Methods: PJI was diagnosed according to the Musculoskeletal Infection Society (MSIS) criteria. The microorganisms were detected by culture, mNGS and tNGS to compare the diagnostic effectiveness of the three methods for PJI and to compare their consistency in detecting microorganisms. Drug resistance genes were detected using tNGS. The costs and turnaround times of mNGS and tNGS were compared. Results: Forty-three patients with PJI, 21 patients without PJI and 10 negative control cases were included. The culture, tNGS, and mNGS sensitivities for PJI diagnosis were 74.41%, 88.37%, and 93.02%, respectively, with no significant differences. The specificities were 90.48%, 95.24%, and 95.24%, respectively, with no significant differences. tNGS detected drug resistance genes in 37.5% of culture-positive PJIs. tNGS was superior to mNGS for turnaround time (14.5 h vs. 28 h) and cost ($150 vs. $260). Conclusions: tNGS can effectively identify PJI pathogens and may provide drug resistance information, while tNGS is superior to mNGS regarding cost and turnaround time. A multidisciplinary, multi-technology based algorithm to diagnose PJI is appropriate. Highlights: 298 microorganisms and 86 drug resistance genes were included in the tNGS panel.Diagnostic efficacy of tNGS is not inferior to that of commonly used indicators.tNGS is superior to mNGS in cost and turnaround time.

Removal of toilet paper fibers from residential wastewater: a life cycle assessment.

Toilet paper has been reported as one of the major insoluble pollutant components in the influent of wastewater treatment plants. Toilet paper fibers contribute to a large production of sewage sludge, resulting in a high treatment cost and high energy consumption. To find energy-efficient, cost-effective, and environment-friendly technologies for fiber removal and resource recovery from wastewater, a life-cycle assessment (LCA) was performed to analyze the wastewater treatment processes, including a sieving process for removing and recovering suspended solids before the biodegradation units. Based on the LCA results, it was estimated that the sieve screening process saved 8.57% of energy consumption. The construction phase of sieving consumed 1.31% energy cost compared with the operation phase. Environmental impact analysis showed that sieving reduced the impacts of climate change, human toxicity, fossil depletion, and particulate matter formation, which reduced the total normalized environmental impacts by 9.46%. The life-cycle analysis of the removal of toilet paper fibers from wastewater revealed the need to use more efficient methods to enhance the recovery of cellulose fibers.

Diversity, functional potential, and assembly of bacterial communities in metal(loid)-contaminated sediments from a coastal industrial basin.

A comprehensive understanding of ecological processes related to bacterial communities in these ecosystems is critical for improving the sustainability of estuarine-coastal ecosystem functions. However, the community composition, functional potential, and assembly mechanisms of bacterial community in metal(loid)-contaminated estuarine-coastal habitats remain poorly understood, especially along lotic systems extending from rivers to estuaries and to bays. Here, we collected sediment samples associated with sewage outlets from rivers (upstream/midstream of sewage outlet), estuaries (sewage outlets), and Jinzhou Bay (downstream of sewage outlets) within Liaoning Province, China in order to evaluate the relationships between microbiome and metal(loid) contamination. Sewage discharge markedly increased the concentrations of metal(loid)s in sediments, including of As, Fe, Co, Pb, Cd, and Zn. Significant discrepancies in alpha diversity and community composition were observed among the sampling sites. The above dynamics were primarily caused by salinity and metal(loid) concentrations (i.e., of As, Zn, Cd, and Pb). Furthermore, metal(loid) stress significantly increased abundances of metal(loid)-resistant genes, but decreased abundances of denitrification genes. Dechloromonas, Hydrogenophaga, Thiobacillus, and Leptothrix were denitrifying bacteria present within sediments of this estuarine-coastal ecosystem. Moreover, the stochastic processes dominated the community assembly in estuary offshore sites, while deterministic processes dominated river community assembly mechanisms. Salinity and total nitrogen concentration, rather than metal(loid) concentrations, governed the assembly processes in these sites. Overall, these elucidate mechanisms involved in constructing community diversities, functional potential, and assembly.

Methadone maintenance treatment alters couplings of default mode and salience networks in individuals with heroin use disorder: A longitudinal self-controlled resting-state fMRI study.

Background: Methadone maintenance treatment (MMT) is a common treatment for heroin use disorder (HUD). Although individuals with HUD have been reported to show impaired coupling among the salience network (SN), executive control network (ECN), and default mode network (DMN), the effects of MMT on the coupling among three large-scale networks in individuals with HUD remains unclear. Methods: Thirty-seven individuals with HUD undergoing MMT and 57 healthy controls were recruited. The longitudinal one-year follow-up study aimed to evaluate the effects of methadone on anxiety, depression, withdrawal symptoms and craving and number of relapse, and brain function (SN, DMN and bilateral ECN) in relation to heroin dependence. The changes in psychological characteristics and the coupling among large-scale networks after 1 year of MMT were analyzed. The associations between the changes in coupling among large-scale networks and psychological characteristics and the methadone dose were also examined. Results: After 1 year of MMT, individuals with HUD showed a reduction in the withdrawal symptom score. The number of relapses was negatively correlated with the methadone dose over 1 year. The functional connectivity between the medial prefrontal cortex (mPFC) and the left middle temporal gyrus (MTG; both key nodes of the DMN) was increased, and the connectivities between the mPFC and the anterior insular and middle frontal gyrus (key nodes of the SN) were also increased. The mPFC-left MTG connectivity was negatively correlated with the withdrawal symptom score. Conclusion: Long-term MMT enhanced the connectivity within the DMN which might be related to reduced withdrawal symptoms, and that between the DMN and SN which might be related to increase in salience values of heroin cues in individuals with HUD. Long-term MMT may be a double-edged sword in treatment for HUD.

Enhanced resistance to Bacillus thuringiensis Cry1Ac toxin mediated by the activation of prophenoloxidase in a cosmopolitan pest.

Plutella xylostella has evolved resistance to Bacillus thuringiensis Cry1Ac toxin over a long evolutionary period. Enhanced immune response is an important factor in insect resistance to a variety of insecticides, and whether phenoloxidase (PO), an immune protein, is involved in resistance to Cry1Ac toxin in P. xylostella remains unclear. Here, spatial and temporal expression patterns showed that prophenoloxidase (PxPPO1 and PxPPO2) in the Cry1S1000-resistant strain was more highly expressed in eggs, 4th instar, head, and hemolymph than those in G88-susceptible strain. The results of PO activity analysis showed that after treatment with Cry1Ac toxin PO activity was about 3 times higher than that before treatment. Furthermore, knockout of PxPPO1 and PxPPO2 significantly increased the susceptibility to Cry1Ac toxin. These findings were further supported by the knockdown of Clip-SPH2, a negative regulator of PO, which resulted in increased PxPPO1 and PxPPO2 expression and Cry1Ac susceptibility in the Cry1S1000-resistant strain. Finally, the synergistic effect of quercetin showed that larval survival decreased from 100 % to <20 % compared to the control group. This study will provide a theoretical basis for the analysis of immune-related genes (PO) genes involved in the resistance mechanism and pest control of P. xylostella.

Deep learning for fully automated segmentation and volumetry of Couinaud liver segments and future liver remnants shown with CT before major hepatectomy: a validation study of a predictive model.

Background: Recent reports have shown the potential for deep learning (DL) models to automatically segment of Couinaud liver segments and future liver remnant (FLR) for liver resections. However, these studies have mainly focused on the development of the models. Existing reports lack adequate validation of these models in diverse liver conditions and thorough evaluation using clinical cases. This study thus aimed to develop and perform a spatial external validation of a DL model for the automated segmentation of Couinaud liver segments and FLR using computed tomography (CT) in various liver conditions and to apply the model prior to major hepatectomy. Methods: This retrospective study developed a 3-dimensional (3D) U-Net model for the automated segmentation of Couinaud liver segments and FLR on contrast-enhanced portovenous phase (PVP) CT scans. Images were obtained from 170 patients from January 2018 to March 2019. First, radiologists annotated the Couinaud segmentations. Then, a 3D U-Net model was trained in Peking University First Hospital (n=170) and tested in Peking University Shenzhen Hospital (n=178) in cases with various liver conditions (n=146) and in candidates for major hepatectomy (n=32). The segmentation accuracy was evaluated using the dice similarity coefficient (DSC). Quantitative volumetry to evaluate the resectability was compared between manual and automated segmentation. Results: The DSC in the test data sets 1 and 2 for segments I to VIII was 0.93±0.01, 0.94±0.01, 0.93±0.01, 0.93±0.01, 0.94±0.00, 0.95±0.00, 0.95±0.00, and 0.95±0.00, respectively. The mean automated FLR and FLR% assessments were 493.51±284.77 mL and 38.53%±19.38%, respectively. The mean manual FLR and FLR% assessments were 500.92±284.38 mL and 38.35%±19.14%, respectively, in test data sets 1 and 2. For test data set 1, when automated segmentation of the FLR% was used, 106, 23, 146, and 57 cases were categorized as candidates for a virtual major hepatectomy of types 1, 2, 3, and 4, respectively; however, when manual segmentation of the FLR% was used, 107, 23, 146, and 57 cases were categorized as candidates for a virtual major hepatectomy of types 1, 2, 3, and 4, respectively. For test data set 2, all cases were categorized as candidates for major hepatectomy when automated and manual segmentation of the FLR% was used. No significant differences in FLR assessment (P=0.50; U=185,545), FLR% assessment (P=0.82; U=188,337), or the indications for major hepatectomy were noted between automated and manual segmentation (McNemar test statistic 0.00; P>0.99). Conclusions: The DL model could be used to fully automate the segmentation of Couinaud liver segments and FLR with CT prior to major hepatectomy in an accurate and clinically practicable manner.

Analysis of the Effect of Plutella xylostella Polycalin and ABCC2 Transporter on Cry1Ac Susceptibility by CRISPR/Cas9-Mediated Knockout.

Many insects, including the Plutella xylostella (L.), have developed varying degrees of resistance to many insecticides, including Bacillus thuringiensis (Bt) toxins, the bioinsecticides derived from Bt. The polycalin protein is one of the potential receptors for Bt toxins, and previous studies have confirmed that the Cry1Ac toxin can bind to the polycalin protein of P. xylostella, but whether polycalin is associated with the resistance of Bt toxins remains controversial. In this study, we compared the midgut of larvae from Cry1Ac-susceptible and -resistant strains, and found that the expression of the Pxpolycalin gene was largely reduced in the midgut of the resistant strains. Moreover, the spatial and temporal expression patterns of Pxpolycalin showed that it was mainly expressed in the larval stage and midgut tissue. However, genetic linkage experiments showed that the Pxpolycalin gene and its transcript level were not linked to Cry1Ac resistance, whereas both the PxABCC2 gene and its transcript levels were linked to Cry1Ac resistance. The larvae fed on a diet containing the Cry1Ac toxin showed no significant change in the expression of the Pxpolycalin gene in a short term. Furthermore, the knockout of polycalin and ATP-binding cassette transporter subfamily C2 (ABCC2) genes separately by CRISPR/Cas9 technology resulted in resistance to decreased susceptibility to Cry1Ac toxin. Our results provide new insights into the potential role of polycalin and ABCC2 proteins in Cry1Ac resistance and the mechanism underlying the resistance of insects to Bt toxins.