Advantages of robot-assisted resection of large mediastinal tumors: a single-center preliminary study.

Current study aims to assess the safety and efficacy of robot-assisted thoracoscopic surgery (RATS) for sizable mediastinal masses with a minimum diameter ≥6 cm, compared with video-assisted thoracoscopic surgery (VATS) and open surgery. This study enrolled 130 patients with mediastinal tumors with no less than 6 cm diameter in Zhongnan Hospital, Wuhan University, including 33 patients who underwent RATS, 52 patients who underwent VATS and 45 patients who underwent open surgery. After classifying based on mass size and whether it has invaded or not, we compared their clinical characteristics and perioperative outcomes. There was no significant difference in age, gender, mass size, myasthenia gravis, mass location, pathological types (p > 0.05) in three groups. Patients undergoing open surgery typically presenting at a more advanced stage (p < 0.05). No obvious difference was discovered in the average postoperative length of stay, operation duration, chest tube duration and average postoperative day 1 drainage output between RATS group and VATS group (p > 0.05), while intraoperative blood loss in RATS group was significantly lower than VATS group (p = 0.046). Moreover, the postoperative length of stay, operation duration, chest tube duration and intraoperative blood loss in RATS group were significantly lower than open surgery group (p < 0.001). RATS is a secure and efficient approach for removing large mediastinal masses at early postoperative period. In comparison with VATS, RATS is associated with lower intraoperative blood loss. Compared with open surgery, RATS is also associated with shorter postoperative length of stay, operation duration, chest tube duration and intraoperative blood loss.

Nano-cutting mechanism of ion implantation-modified SiC: reducing subsurface damage expansion and abrasive wear.

This study utilized ion implantation to modify the material properties of silicon carbide (SiC) to mitigate subsurface damage during SiC machining. The paper analyzed the mechanism of hydrogen ion implantation on the machining performance of SiC at the atomic scale. A molecular dynamics model of nanoscale cutting of an ion-implanted SiC workpiece using a non-rigid regular tetrakaidecahedral diamond abrasive grain was established. The study investigated the effects of ion implantation on crystal structure phase transformation, dislocation nucleation, and defect structure evolution. Results showed ion implantation modification decreased the extension depth of amorphous structures in the subsurface layer, thereby enhancing the surface and subsurface integrity of the SiC workpiece. Additionally, dislocation extension length and volume within the lattice structure were lower in the ion-implanted workpiece compared to non-implanted ones. Phase transformation, compressive pressure, and cutting stress of the lattice in the shear region per unit volume were lower in the ion-implanted workpiece than the non-implanted one. Taking the diamond abrasive grain as the research subject, the mechanism of grain wear under ion implantation was explored. Grain expansion, compression, and atomic volumetric strain wear rate were higher in the non-implanted workpiece versus implanted ones. Under shear extrusion of the SiC workpiece, dangling bonds of atoms in the diamond grain were unstable, resulting in graphitization of the diamond structure at elevated temperatures. This study established a solid theoretical and practical foundation for realizing non-destructive machining at the atomic scale, encompassing both theoretical principles and practical applications. .

Discovery of Near-Infrared Heptamethine Cyanine Probes for Imaging-Guided Surgery in Solid Tumors.

Near-infrared (NIR) fluorescence imaging has attracted much attention in image-guided interventions with unique advantages. However, the clinical translation rate of fluorescence probes is extremely low, primarily due to weak lesion signal contrast and poor specificity. To address this dilemma, a series of small-molecule near-infrared fluorescence probes have been designed for tumor imaging. Among them, YQ-04-03 showed notable optical stability and remarkable sensitivity toward tumor targeting. Moreover, within a specific concentration and time range against oxidizing reducing agents and laser, it demonstrated better stability than ICG. The retention time of YQ-04-03 in tumors was significantly longer compared to other nonspecific uptake sites in the subjects, and its tumor-to-normal tissue ratio (TNR) outperformed ICG. Successful resection of in situ hepatocarcinoma and peritoneal carcinoma was achieved using probe imaging guidance, with the smallest visual lesion resected measuring approximately 1 mm3. Ultimately, this probe holds great potential for advancing tumor tracer.

High-density bin-based genetic map reveals a 530-kb chromosome segment derived from wild peanut contributing to late leaf spot resistance.

KEY MESSAGE: Twenty-eight QTLs for LLS disease resistance were identified using an amphidiploid constructed mapping population, a favorable 530-kb chromosome segment derived from wild species contributes to the LLS resistance. Late leaf spot (LLS) is one of the major foliar diseases of peanut, causing serious yield loss and affecting the quality of kernel and forage. Some wild Arachis species possess higher resistance to LLS as compared with cultivated peanut; however, ploidy level differences restrict utilization of wild species. In this study, a synthetic amphidiploid (Ipadur) of wild peanuts with high LLS resistance was used to cross with Tifrunner to construct TI population. In total, 200 recombinant inbred lines were collected for whole-genome resequencing. A high-density bin-based genetic linkage map was constructed, which includes 4,809 bin markers with an average inter-bin distance of 0.43 cM. The recombination across cultivated and wild species was unevenly distributed, providing a novel recombination landscape for cultivated-wild Arachis species. Using phenotyping data collected across three environments, 28 QTLs for LLS disease resistance were identified, explaining 4.35-20.42% of phenotypic variation. The major QTL located on chromosome 14, qLLS14.1, could be consistently detected in 2021 Jiyang and 2022 Henan with 20.42% and 12.12% PVE, respectively. A favorable 530-kb chromosome segment derived from Ipadur was identified in the region of qLLS14.1, in which 23 disease resistance proteins were located and six of them showed significant sequence variations between Tifrunner and Ipadur. Allelic variation analysis indicating the 530-kb segment of wild species might contribute to the disease resistance of LLS. These associate genomic regions and candidate resistance genes are of great significance for peanut breeding programs for bringing durable resistance through pyramiding such multiple LLS resistance loci into peanut cultivars.

A Respiratory Motion Prediction Method Based on LSTM-AE with Attention Mechanism for Spine Surgery.

Respiratory motion-induced vertebral movements can adversely impact intraoperative spine surgery, resulting in inaccurate positional information of the target region and unexpected damage during the operation. In this paper, we propose a novel deep learning architecture for respiratory motion prediction, which can adapt to different patients. The proposed method utilizes an LSTM-AE with attention mechanism network that can be trained using few-shot datasets during operation. To ensure real-time performance, a dimension reduction method based on the respiration-induced physical movement of spine vertebral bodies is introduced. The experiment collected data from prone-positioned patients under general anaesthesia to validate the prediction accuracy and time efficiency of the LSTM-AE-based motion prediction method. The experimental results demonstrate that the presented method (RMSE: 4.39%) outperforms other methods in terms of accuracy within a learning time of 2 min. The maximum predictive errors under the latency of 333 ms with respect to the x, y, and z axes of the optical camera system were 0.13, 0.07, and 0.10 mm, respectively, within a motion range of 2 mm.

Environmental levels of azoxystrobin disturb male zebrafish behavior: Possible roles of oxidative stress, cholinergic system, and dopaminergic system.

A widely applied pesticide of azoxystrobin, is increasingly detected in the water environment. Concern has been raised against its potential detriment to aquatic ecosystems. It has been shown that exposure to azoxystrobin interfere with the locomotor behavior of zebrafish larvae. This study aims to investigate whether exposure to environmental levels of azoxystrobin (2 µg/L, 20 µg/L, and 200 µg/L) changes the behavior of male adult zebrafish. Herein, we evaluated behavioral response (locomotor, anxiety-like, and exploratory behaviors), histopathology, biochemical indicators, and gene expression in male adult zebrafish upon azoxystrobin exposure. The study showed that exposure to azoxystrobin for 42 days remarkably increased the locomotor ability of male zebrafish, resulted in anxiety-like behavior, and inhibited exploratory behavior. After treatment with 200 µg/L azoxystrobin, vasodilatation, and congestion were observed in male zebrafish brains. Exposure to 200 µg/L azoxystrobin notably elevated ROS level, MDA concentration, CAT activity, and AChE activity, while inhibiting SOD activity, GPx activity, ACh concentration, and DA concentration in male zebrafish brains. Moreover, the expression levels of genes related to the antioxidant, cholinergic, and dopaminergic systems were significantly changed. This suggests that azoxystrobin may interfere with the homeostasis of neurotransmitters by causing oxidative stress in male zebrafish brains, thus affecting the behavioral response of male zebrafish.

Integrating transcriptomics and network pharmacology to reveal the mechanisms of total Rhizoma Coptidis alkaloids against nonalcoholic steatohepatitis.

ETHNOPHARMACOLOGICAL RELEVANCE: Non-alcoholic steatohepatitis (NASH) has emerged as a major cause of cirrhosis and hepatocellular carcinoma, posing a significant threat to public health. Rhizoma Coptidis, a traditional Chinese medicinal herb has been shown to have significant curative effects on liver diseases. Total Rhizoma Coptidis Alkaloids (TRCA) is a primarily alkaloid mixture extracted from Rhizoma Coptidis, and its constituents are widely accepted to have hepatoprotective effects. AIM OF THE STUDY: This work aimed to investigate the efficacy and potential mechanisms of TRCA in ameliorating NASH through both in vitro experiments and in vivo mouse models. MATERIALS AND METHODS: The study employed a mice model induced by a high-fat diet (HFD) to evaluate the effectiveness and pharmacological mechanisms of TRCA in alleviating NASH. Transcriptomic sequencing and network pharmacology were used to explore the possible targets and mechanisms of TRCA to ameliorate NASH. Further validation was performed in free fatty acid (FFA)-induced human hepatocytes (LO2) and human hepatocellular carcinoma cells (HepG2). RESULTS: TRCA effectively ameliorated the main features of NASH such as lipid accumulation, hepatitis and hepatic fibrosis in the liver tissue of mice induced by HFD, as well as improved glucose tolerance and insulin resistance in mice. Combined with transcriptomic and network pharmacological analyses, 68 core targets associated with the improvement of NASH by TRCA were obtained. According to the KEGG results, the core targets were significantly enriched in the PI3K-AKT signaling pathway whereas TRCA ameliorated the aberrant down-regulation of the PI3K-AKT signaling pathway induced by HFD. Furthermore, the five highest-ranked genes were obtained by PPI network analysis. Moreover, our findings suggest that TRCA may impede the progression of HFD-induced NASH by regulating the expression of PPARG, MMP9, ALB, CCL2, and EGFR. CONCLUSIONS: TRCA can ameliorate HFD-induced liver injury by modulating aberrant downregulation of the PI3K-AKT signaling pathway. Key proteins such as PPARG, MMP9, ALB, CCL2, and EGFR may be critical targets for TRCA to ameliorate NASH. This finding supports using Rhizoma Coptidis, a well-known herbal medicine, as a potential therapeutic agent for NASH.

Multiprior Learning via Neural Architecture Search for Blind Face Restoration.

Blind face restoration (BFR) aims to recover high-quality (HQ) face images from low-quality (LQ) ones and usually resorts to facial priors for improving restoration performance. However, current methods still suffer from two major difficulties: 1) how to derive a powerful network architecture without extensive hand tuning and 2) how to capture complementary information from multiple facial priors in one network to improve restoration performance. To this end, we propose a face restoration searching network (FRSNet) to adaptively search the suitable feature extraction architecture within our specified search space, which can directly contribute to the restoration quality. On the basis of FRSNet, we further design our multiple facial prior searching network (MFPSNet) with a multiprior learning scheme. MFPSNet optimally extracts information from diverse facial priors and fuses the information into image features, ensuring that both external guidance and internal features are reserved. In this way, MFPSNet takes full advantage of semantic-level (parsing maps), geometric-level (facial heat maps), reference-level (facial dictionaries), and pixel-level (degraded images) information and, thus, generates faithful and realistic images. Quantitative and qualitative experiments show that the MFPSNet performs favorably on both synthetic and real-world datasets against the state-of-the-art (SOTA) BFR methods. The codes are publicly available at: https://github.com/YYJ1anG/MFPSNet.

Effect of orthopedic insoles on spinal deformity and walking in adolescents with idiopathic scoliosis summary.

Objective: To observe the effects of scoliosis-specific exercise therapy combined with braces and orthopedic insoles on improved spinal deformity and walking ability in adolescents with idiopathic scoliosis (AIS). Method: From September 2019 to September 2020, 60 outpatient AIS patients were distributed into brace group (n = 30) at random and brace combined orthopedic insole group (n = 30). Both groups underwent brace dryness, and the observation group used scoliosis-specific exercise therapy combined with brace therapy, and on this basis, orthopedic insole intervention was added for 8 h per day for 2 months. At the same time, 20 adolescents of the same age with normal spinal development were recruited as a healthy group. GaitScan instruments were used to collect gait and plantar pressure measurements from study subjects. First, the gait and plantar pressure data of AIS patients and healthy groups were compared horizontally to ascertain the abnormal indicators, and then the spinal deformity and the above abnormal indicators were compared between the brace group and the brace combined orthopedic insole group. Outcome: The plantar pressure center drift index (CPEI) in the AIS group was higher than that in the healthy group (F = 3.120, P < 0.05), and there were significant differences in the ratio of medial and lateral heel pressure (M/l) and total foot pressure (P < 0.05) between the AIS group and the healthy group, and no noticeable variations were found in the support phase period, walking speed, and proportion of each phase (P > 0.05). After treatment, the Cobb angle was significantly reduced in both the brace group and the brace combined with orthopedic insole group (P < 0.05), and there was no significant difference between the groups (P > 0.05). There were no significant changes in the pressure ratio of CPEI, M/l and bilateral full foot in the brace group (P > 0.05). The CPEI decreased in the brace combined with orthopedic insole group (P < 0.05), and the pressure ratio of M/l and bilateral full foot tended to 1 (P < 0.05), and was better than that in the brace group (P < 0.05). Conclusion: Patients with AIS may have local and worldwide asymmetric changes in plantar pressure distribution. The addition of orthopedic insoles has limited effect on improving scoliosis deformity in patients with AIS, but it can effectively improve the abnormal biomechanics of patients with AIS, so that the patient's force tends to be balanced.

Phylogenomic Analysis of Cytochrome P450 Gene Superfamily and Their Association with Flavonoids Biosynthesis in Peanut (Arachis hypogaea L.).

Cytochrome P450s (CYPs) constitute extensive enzyme superfamilies in the plants, playing pivotal roles in a multitude of biosynthetic and detoxification pathways essential for growth and development, such as the flavonoid biosynthesis pathway. However, CYPs have not yet been systematically studied in the cultivated peanuts (Arachis hypogaea L.), a globally significant cash crop. This study addresses this knowledge deficit through a comprehensive genome-wide analysis, leading to the identification of 589 AhCYP genes in peanuts. Through phylogenetic analysis, all AhCYPs were systematically classified into 9 clans, 43 gene families. The variability in the number of gene family members suggests specialization in biological functions. Intriguingly, both tandem duplication and fragment duplication events have emerged as pivotal drivers in the evolutionary expansion of the AhCYP superfamily. Ka/Ks analysis underscored the substantial influence of strong purifying selection on the evolution of AhCYPs. Furthermore, we selected 21 genes encoding 8 enzymes associated with the flavonoid pathway. The results of quantitative real-time PCR (qRT-PCR) experiments unveiled stage-specific expression patterns during the development of peanut testa, with discernible variations between pink and red testa. Importantly, we identified a direct correlation between gene expression levels and the accumulation of metabolites. These findings offer valuable insights into elucidating the comprehensive functions of AhCYPs and the underlying mechanisms governing the divergent accumulation of flavonoids in testa of different colors.

Simulation and Experimental Verification of Magnetic Field Diffusion at the Launch Load during Electromagnetic Launch.

The unique magnetic field environment during electromagnetic launch imposes higher requirements on the design and protection of the internal electronic system within the launch load. This low-frequency, Tesla-level extreme magnetic field environment is fundamentally distinct from the Earth's geomagnetic field. The excessive change rate of magnetic flux can readily induce voltage within the circuit, thus disrupting the normal operation of intelligent microchips. Existing simulation methods primarily focus on the physical environments of rails and armatures, making it challenging to precisely compute the magnetic field environment at the load's location. In this paper, we propose a computational rail model based on the magneto-mechanical coupling model of a railgun. This model accounts for the dynamic current distribution during the launch process and simulates the magnetic flux density distribution at the load location. To validate the model's accuracy, three-axis magnetic sensors were placed in front of the armature, and the dynamic magnetic field distribution during the launch process was obtained using the projectile-borne-storage testing method. The results indicate that compared to the previous literature methods, the approach proposed in this paper achieves higher accuracy and is closer to experimental results, providing valuable support for the design and optimization of the launch load.

Afforestation changed the fungal functional community of paddy fields and dry farmlands differently.

Afforestation currently makes a great contribution to carbon uptake in terrestrial ecosystems, while dramatically affects soil ecosystem functions too. Little is known, however, about the changes in soil fungal functional groups and their interactions following afforestation. Here, based on high-throughput sequencing and FUNGuild annotation, we investigated the functional characteristics of soil fungi as well as environmental factors in a watershed where paddy field and dry farmland were changed to eucalyptus plantation. The results showed that afforestation on paddy field resulted in greater changes in diversity, community structure and taxon interactions of fungal functional groups than afforestation on dry farmland. The most complex and distinctive community structure was found in eucalyptus plantation, as well as the greatest taxon interactions, and the lowest alpha-diversity of functional guilds of symbiotrophic fungi because of the dominant ectomycorrhizal fungi. Paddy field exhibited the highest proportion of saprotrophic fungi, but the lowest taxonomic diversity of saprotrophic and pathotrophic fungi. The taxonomic diversity of undefined saprotrophic fungi shaped the differences in community structure and network complexity between eucalyptus plantation and cropland. Limited cooperation within dominant fungi was the main reason for the establishment of a loose co-occurrence network in paddy field. From croplands to artificial forests, reduced soil pH boosted the taxonomic diversity of fungal functional groups. All of these findings suggested that afforestation may lead to an increase in the taxonomic diversity of soil fungal functional groups, which would further intensify the taxon interactions.

Online semi-supervised learning for motor imagery EEG classification.

OBJECTIVE: Time-consuming data labeling in brain-computer interfaces (BCIs) raises many problems such as mental fatigue and is one key factor that hinders the real-world adoption of motor imagery (MI)-based BCIs. An alternative approach is to integrate readily available, as well as informative, unlabeled data online, whereas this approach is less investigated. APPROACH: We proposed an online semi-supervised learning scheme to improve the classification performance of MI-based BCI. This scheme uses regularized weighted online sequential extreme learning machine (RWOS-ELM) as the base classifier and updates its model parameters with incoming balanced data chunk-by-chunk. In the initial stage, we designed a technique that combines the synthetic minority oversampling with the edited nearest neighbor rule for data augmentation to construct more discriminative initial classifiers. When used online, the incoming chunk of data is first pseudo-labeled by RWOS-ELM as well as an auxiliary classifier, and then balanced again by the above-mentioned technique. Initial classifiers are further updated based on these class-balanced data. MAIN RESULTS: Offline experimental results on two publicly available MI datasets demonstrate the superiority of the proposed scheme over its counterparts. Further online experiments on six subjects show that their BCI performance gradually improved by learning from incoming unlabeled data. SIGNIFICANCE: Our proposed online semi-supervised learning scheme has higher computation and memory usage efficiency, which is promising for online MI-based BCIs, especially in the case of insufficient labeled training data.

Circulating exosomal microRNA-4497 as a potential biomarker for metastasis and prognosis in non-small-cell lung cancer.

Circulating exosomal microRNAs (miRNAs) have shown great potential for the diagnosis, prognosis, and treatment monitoring of patients with non-small-cell lung cancer (NSCLC). Our main purpose was to determine the clinical value of serum exosomal miR-4497 as a new non-invasive biomarker for NSCLC. The exoRNeasy Kit (QIAGEN, Hilden, Germany) was used to isolate exosomes and exoRNA from the serum of 84 patients with NSCLC (NSCLC group), 30 patients with benign lung lesion (BLL group), and 47 healthy controls. Six serum exosomal miRNAs (Let-7b-5p, miR-122-5p, miR-155-5p, miR-223-3p, miR-320c, and miR-4497) were selected as candidate miRNAs and analyzed using real-time qPCR, among which miR-4497 displayed the most striking differences. Exosomal miR-4497 expressed significantly lower in NSCLC than in BLL patients and healthy controls (P < 0.001). Further investigation showed that miR-4497 was negatively correlated with the malignant characteristics of tumors (tumor size, tumor-node-metastasis [TNM] stage, and distant metastasis) and was an independent tumor suppressor (P < 0.05). According to receiver operating characteristic (ROC) analysis, exosomal miR-4497 independently exhibited excellent diagnostic efficacy, which could be improved by combining it with traditional markers (for identifying tumor size, the area under the curve [AUC] = 0.761; TNM stage, AUC = 0.878; distant metastasis, AUC = 0.895; all P < 0.001). Moreover, longitudinal analysis revealed that exosomal miR-4497 levels increased after chemoradiotherapy (P < 0.001). According to the survival analysis, poor overall survival (OS) and disease-free survival (DFS) were associated with low exosomal miR-4497 levels (P < 0.05). Moreover, exosomal miR-4497 was an independent protective factor affecting DFS (hazard ratio = 0.190, P = 0.009) in the Cox proportional hazards model. Therefore, serum exosomal miR-4497 can be used as a potential biomarker to identify NSCLC and healthy individuals or BLL patients for early screening or as a biomarker for staging and grading, prognosis, and monitoring recurrence, metastasis, and the therapeutic effects in patients with NSCLC.

Fixed-Time Recurrent NN Learning Control of Uncertain Robotic Manipulators with Time-Varying Constraints: Experimental Verification.

This paper proposes a learning control framework for the robotic manipulator's dynamic tracking task demanding fixed-time convergence and constrained output. In contrast with model-dependent methods, the proposed solution deals with unknown manipulator dynamics and external disturbances by virtue of a recurrent neural network (RNN)-based online approximator. First, a time-varying tangent-type barrier Lyapunov function (BLF) is introduced to construct a fixed-time virtual controller. Then, the RNN approximator is embedded in the closed-loop system to compensate for the lumped unknown term in the feedforward loop. Finally, we devise a novel fixed-time, output-constrained neural learning controller by integrating the BLF and RNN approximator into the main framework of the dynamic surface control (DSC). The proposed scheme not only guarantees the tracking errors converge to the small neighborhoods about the origin in a fixed time, but also preserves the actual trajectories always within the prescribed ranges and thus improves the tracking accuracy. Experiment results illustrate the excellent tracking performance and verify the effectiveness of the online RNN estimate for unknown dynamics and external disturbances.

Longitudinal Impact of Perfectionism on Suicidal Ideation among Chinese College Students with Perceived Academic Failure: The Roles of Rumination and Depression.

OBJECTIVE: Suicide has become a nonnegligible cause of death among emerging adults, and academic performance is one of the most significant factors in Chinese college students' suicidal ideation. Based on this, we examined the risk and protective sides of perfectionism on suicidal ideation among college students with perceived academic failure experiences. METHOD: In the current study, a total of 674 Chinese college students (63.6% females, Mage = 19.96, SD = 1.31) with perceived academic failure in the past six months participated in a survey and completed self-report measures assessing perfectionism, rumination, depression, and suicidal ideation twice with a six-month interval. RESULTS: Negative perfectionism was a risk factor for suicidal ideation, while positive perfectionism had an opposite effect. In addition, intrusive rumination, deliberate rumination, and depression serially mediated the association between positive/negative perfectionism and suicidal ideation. CONCLUSION: The results indicated perfectionism has both positive and negative sides to suicidal ideation. Clinicians might be aware of the risk and protective components of perfectionism when intervening with suicidal ideation among college students with perceived academic failure experiences.

The current study examined the longitudinal association between negative/positive perfectionism and suicidal ideation among college students with perceived academic failure.Negative perfectionism generally acted as a risk factor for suicidal ideation, while positive perfectionism showed the opposite effects.he serial mediation among perfectionism, rumination, depression, and suicidal ideation was formed.

Imaging, Genetic, and Pathological Features of Vascular Dementia.

BACKGROUND: Over the past decades, marked progress has been made in detecting vascular dementia (VD) both through maturation of diagnostic concepts and advances in brain imaging, especially MRI. We summarized the imaging, genetic, and pathological features of VD in this review. SUMMARY: It is a challenge for the diagnosis and treatment of VD, particularly in patients where there is no evident temporal relation between cerebrovascular events and cognitive dysfunction. In patients with cognitive dysfunction with poststroke onset, the etiological classification is still complicated. KEY MESSAGES: In this review, we summarized the clinical, imaging, and genetic as well as pathological features of VD. We hope to offer a framework to translate diagnostic criteria to daily practice, address treatment, and highlight some future perspectives.

In Vivo Staging the Progression of Colitis and Associated Cancer by Concurrent Microimaging of Key Biomarkers.

Currently colorectal cancer (CRC) staging (colitis, adenoma, and carcinoma) mainly relies on ex vivo pathologic analysis requiring an invasive surgical process with limited sample collection and increased metastatic risk. Thus, in vivo noninvasive pathological diagnosis is extremely demanded. By verifying the samples of clinical patients and CRC mouse models, it was found that vascular endothelial growth factor receptor 2 (VEGFR2) was barely expressed in the colitis stage and only appeared in adenoma and carcinoma stages with obvious elevation, while prostaglandin E receptor 4 (PTGER4) could be observed from colitis to adenoma and carcinoma stages with a gradient increase of expression. VEGFR2 and PTGER4 were further chosen as key biomarkers for molecular pathological diagnosis in vivo and corresponding molecular probes were constructed. The feasibility of in vivo noninvasive CRC staging by concurrent microimaging of dual biomarkers using confocal laser endoscopy (CLE) was verified in CRC mouse models and further confirmed by ex vivo pathological analysis. In vivo CLE imaging exhibited the correlation of severe colonic crypt structural alteration with a higher biomarker expression in adenoma and carcinoma stages. This strategy shows promise in benefiting patients undergoing CRC progression with in-time, noninvasive, and precise pathological staging, thus providing valuable guidance for selecting therapeutic strategies.

Tomato geranylgeranyl diphosphate synthase isoform 1 is involved in the stress-triggered production of diterpenes in leaves and strigolactones in roots.

Carotenoids are photoprotectant pigments and precursors of hormones such as strigolactones (SL). Carotenoids are produced in plastids from geranylgeranyl diphosphate (GGPP), which is diverted to the carotenoid pathway by phytoene synthase (PSY). In tomato (Solanum lycopersicum), three genes encode plastid-targeted GGPP synthases (SlG1 to SlG3) and three genes encode PSY isoforms (PSY1 to PSY3). Here, we investigated the function of SlG1 by generating loss-of-function lines and combining their metabolic and physiological phenotyping with gene co-expression and co-immunoprecipitation analyses. Leaves and fruits of slg1 lines showed a wild-type phenotype in terms of carotenoid accumulation, photosynthesis, and development under normal growth conditions. In response to bacterial infection, however, slg1 leaves produced lower levels of defensive GGPP-derived diterpenoids. In roots, SlG1 was co-expressed with PSY3 and other genes involved in SL production, and slg1 lines grown under phosphate starvation exuded less SLs. However, slg1 plants did not display the branched shoot phenotype observed in other SL-defective mutants. At the protein level, SlG1 physically interacted with the root-specific PSY3 isoform but not with PSY1 and PSY2. Our results confirm specific roles for SlG1 in producing GGPP for defensive diterpenoids in leaves and carotenoid-derived SLs (in combination with PSY3) in roots.

Blocking autofluorescence in brain tissues affected by ischemic stroke, hemorrhagic stroke, or traumatic brain injury.

Autofluorescence is frequently observed in animal tissues, interfering with an experimental analysis and leading to inaccurate results. Sudan black B (SBB) is a staining dye widely used in histological studies to eliminate autofluorescence. In this study, our objective was to characterize brain tissue autofluorescence present in three models of acute brain injury, including collagenase-induced intracerebral hemorrhage (ICH), traumatic brain injury (TBI), and middle cerebral artery occlusion, and to establish a simple method to block autofluorescence effectively. Using fluorescence microscopy, we examined autofluorescence in brain sections affected by ICH and TBI. In addition, we optimized a protocol to block autofluorescence with SBB pretreatment and evaluated the reduction in fluorescence intensity. Compared to untreated, pretreatment with SBB reduced brain tissue autofluorescence in the ICH model by 73.68% (FITC), 76.05% (Tx Red), and 71.88% (DAPI), respectively. In the TBI model, the ratio of pretreatment to untreated decreased by 56.85% (FITC), 44.28% (Tx Red), and 46.36% (DAPI), respectively. Furthermore, we tested the applicability of the protocol using immunofluorescence staining or Cyanine-5.5 labeling in the three models. SBB treatment is highly effective and can be applied to immunofluorescence and fluorescence label imaging techniques. SBB pretreatment effectively reduced background fluorescence but did not significantly reduce the specific fluorescence signal and greatly improved the signal-to-noise ratio of fluorescence imaging. In conclusion, the optimized SBB pretreatment protocol blocks brain section autofluorescence of the three acute brain injury models.