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Global climate change is affecting plant photosynthesis and transpiration processes, as well as increasing weather extremes impacting socio-political and environmental events and decisions for decades to come. One major research challenge in plant biology and ecology is the interaction of photosynthesis with the environment. Stomata control plant gas exchange and their evolution was a crucial innovation that facilitated the earliest land plants to colonize terrestrial environments. Stomata couple homoiohydry, together with cuticles, intercellular gas space, with the endohydric water-conducting system, enabling plants to adapt and diversify across the planet. Plants control stomatal movement in response to environmental change through regulating guard cell turgor mediated by membrane transporters and signaling transduction. However, the origin, evolution, and active control of stomata remain controversial topics. We first review stomatal evolution and diversity, providing fossil and phylogenetic evidence of their origins. We summarize functional evolution of guard cell membrane transporters in the context of climate changes and environmental stresses. Our analyses show that the core signaling elements of stomatal movement are more ancient than stomata, while genes involved in stomatal development co-evolved de novo with the earliest stomata. These results suggest that novel stomatal development-specific genes were acquired during plant evolution, whereas genes regulating stomatal movement, especially cell signaling pathways, were inherited ancestrally and co-opted by dynamic functional differentiation. These two processes reflect the different adaptation strategies during land plant evolution., (© 2024 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Introduction: Insulin resistance (IR) plays an important role in the occurrence and development of cardiovascular disease (CVD) in patients with chronic kidney disease (CKD). The triglyceride-glucose (TyG) index is a simple and effective tool to evaluate IR. This study aimed to evaluate the association of the TyG index with coronary artery disease (CAD) and the severity of coronary artery stenosis (CAS) in nondialysis patients with stages 3-5 CKD., Methods: Nondialysis patients with stages 3-5 CKD who underwent the first coronary angiography at Zhongda Hospital affiliated with Southeast University from August 2015 to January 2017 were retrospectively analyzed. CAS was measured by coronary angiography, and the CAS score was calculated as the Gensini score. Logistic regression analysis was used to determine the related factors of CAD and severe CAS., Results: A total of 943 patients were enrolled in this cross-sectional study and 720 (76.4%) of these patients were diagnosed with CAD. The TyG index in the CAD group (7.29 ± 0.63) was significantly higher than that in the non-CAD group (7.11 ± 0.61) ( p  < 0.001). Multivariate logistic regression analysis showed that a higher TyG index was an independent risk factor for CAD in CKD patients after adjusting for related confounding factors (OR = 2.865, 95% CI 1.681-4.885, p  < 0.001). Patients in the CAD group were divided into three groups according to the Gensini integral quantile level. Multivariate logistic regression analysis showed that the TyG index was an independent related factor for severe CAS after adjusting for relevant confounding factors ( p  < 0.001)., Conclusions: The TyG index is associated with CAD and the severity of CAS in patients with nondialysis stages 3-5 CKD. A higher TyG index is an independent factor for CAD and severe CAS.

Reactive oxygen species (ROS) are the key players in regulating developmental processes of plants. Plants have evolved a large array of gene families to facilitate the ROS-regulated developmental process in roots and leaves. However, the cellular targets of ROS during plant evolutionary development are still elusive. Here, we found early evolution and large expansions of protein families such as mitogen-activated protein kinases (MAPK) in the evolutionarily important plant lineages. We review the recent advances in interactions among ROS, phytohormones, gasotransmitters, and protein kinases. We propose that these signaling molecules act in concert to maintain cellular ROS homeostasis in developmental processes of root and leaf to ensure the fine-tuning of plant growth for better adaptation to the changing climate., Competing Interests: Declaration of interests No interests are declared., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A sporadic occurrence of Fanconi syndrome associated with adefovir dipivoxil (ADV) has been reported, particularly when confirmed by renal biopsy. This study presents the case of a 53-year-old man who had been taking ADV 10 mg daily for 10 years to treat chronic hepatitis B (CHB) and subsequently developed Fanconi syndrome. The clinical manifestations included hypophosphatemic osteomalacia, glucosuria, renal tubular acidosis, low-molecular-weight proteinuria, and renal insufficiency. Renal biopsy revealed significant injury to proximal tubular epithelial cells, including vacuolar degeneration and regeneration of tubular epithelial cells. The ultrastructural pathology indicated severe morphological abnormalities of mitochondria, such as densely packed and enlarged mitochondria, with loss, blunting, and disordered arrangement of cristae. Following discontinuation of ADV and supplementation with oral phosphate, hypophosphatemia, glucosuria, and proteinuria were resolved. These findings support the previous hypothesis that ADV-induced nephrotoxicity may involve mitochondrial injury.

Wheat (Triticum aestivum) is one of the most essential human energy and protein sources. However, wheat production is threatened by devastating fungal diseases such as stripe rust, caused by Puccinia striiformis Westend. f. sp. tritici (Pst). Here, we reveal that the alternations in chloroplast lipid profiles and the accumulation of jasmonate (JA) in the necrosis region activate JA signaling and trigger the host defense. The collapse of chloroplasts in the necrosis region results in accumulations of polyunsaturated membrane lipids and the lipid-derived phytohormone JA in transgenic lines of Yr36 that encodes Wheat Kinase START 1 (WKS1), a high-temperature-dependent adult plant resistance protein. WKS1.1, a protein encoded by a full-length splicing variant of WKS1, phosphorylates and enhances the activity of keto-acyl thiolase (KAT-2B), a critical enzyme catalyzing the β-oxidation reaction in JA biosynthesis. The premature stop mutant, kat-2b, accumulates less JA and shows defects in the host defense against Pst. Conversely, overexpression of KAT-2B results in a higher level of JA and limits the growth of Pst. Moreover, JA inhibits the growth and reduces pustule densities of Pst. This study illustrates the WKS1.1‒KAT-2B‒JA pathway for enhancing wheat defense against fungal pathogens to attenuate yield loss., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2023 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.)

Agricultural soil pollution with potentially toxic trace elements (PTEs) has emerged as a significant environmental concern, jeopardizing food safety and human health. Although, conventional remediation approaches have been used for PTEs-contaminated soils treatment; however, these techniques are toxic, expensive, harmful to human health, and can lead to environmental contamination. Nano-enabled agriculture has gained significant attention as a sustainable approach to improve crop production and food security. Silicon nanomaterials (SiNMs) have emerged as a promising alternative for PTEs-contaminated soils remediation. SiNMs have unique characteristics, such as higher chemical reactivity, higher stability, greater surface area to volume ratio and smaller size that make them effective in removing PTEs from the environment. The review discusses the recent advancements and developments in SiNMs for the sustainable remediation of PTEs in agricultural soils. The article covers various synthesis methods, characterization techniques, and the potential mechanisms of SiNMs to alleviate PTEs toxicity in plant-soil systems. Additionally, we highlight the potential benefits and limitations of SiNMs and discusses future directions for research and development. Overall, the use of SiNMs for PTEs remediation offers a sustainable platform for the protection of agricultural soils and the environment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

WUSCHEL (WUS) is a crucial transcription factor in regulating plant stem cell development, and its expression can also improve genetic transformation. However, the ectopic expression of WUS always causes pleiotropic effects during genetic transformation, making it important to understand the regulatory mechanisms underlying these phenomena. In our study, we found that the transient expression of the maize WUS ortholog ZmWus2 caused severe leaf necrosis in Nicotiana benthamiana . We performed transcriptomic and non-target metabolomic analyses on tobacco leaves during healthy to wilted states after ZmWus2 transient overexpression. Transcriptomic analysis revealed that ZmWus2 transformation caused active metabolism of inositol trisphosphate and glycerol-3-phosphate, while also upregulating plant hormone signaling and downregulating photosystem and protein folding pathways. Metabolomic analysis mainly identified changes in the synthesis of phenylpropanoid compounds and various lipid classes, including steroid synthesis. In addition, transcription factors such as ethylene-responsive factors (ERFs), the basic helix-loop-helix (bHLH) factors, and MYBs were found to be regulated by ZmWus2 . By integrating these findings, we developed a WUS regulatory model that includes plant hormone accumulation, stress responses, lipid remodeling, and leaf necrosis. Our study sheds light on the mechanisms underlying WUS ectopic expression causing leaf necrosis and may inform the development of future genetic transformation strategies.

Salvia miltiorrhiza and S. grandifolia are rich in diterpenoids and have therapeutic effects on cardiovascular diseases. In this study, the spatial distribution of diterpenoids in both species was analyzed by a combination of metabolomics and mass spectrometry imaging techniques. The results indicated that diterpenoids in S. miltiorrhiza were mainly abietane-type norditerpenoid quinones with a furan or dihydrofuran D-ring and were mainly distributed in the periderm of the roots, e.g. cryptotanshinone and tanshinone IIA. The compounds in S. grandifolia were mainly phenolic abietane-type tricyclic diterpenoids with six- or seven-membered C-rings, and were widely distributed in the periderm, phloem, and xylem of the roots, e.g. 11-hydroxy-sugiol, 11,20-dihydroxy-sugiol, and 11,20-dihydroxy-ferruginol. In addition, the leaves of S. grandifolia were rich in tanshinone biosynthesis precursors, such as 11-hydroxy-sugiol, while those of S. miltiorrhiza were rich in phenolic acids. Genes in the upstream pathway of tanshinone biosynthesis were highly expressed in the root of S. grandifolia , and genes in the downstream pathway were highly expressed in the root of S. miltiorrhiza . Here, we describe the specific tissue distributions and mechanisms of diterpenoids in two Salvia species, which will facilitate further investigations of the biosynthesis of diterpenoids in plant synthetic biology., Competing Interests: The authors declare that they have no conflicts of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of Nanjing Agricultural University.)

Crop domestication has resulted in nutrient losses, so evaluating the reshaping of phytonutrients is crucial for improving nutrition. Soybean is an ideal model due to its abundant phytonutrients and wild relatives. In order to unravel the domestication consequence of phytonutrients, comparative and association analyses of metabolomes and antioxidant activities were performed on seeds of six wild ( Glycine soja (Sieb. and Zucc.)) and six cultivated soybeans ( Glycine max (L.) Merr.). Through ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), we observed a greater metabolic diversity in wild soybeans, which also displayed higher antioxidant activities. (-)-Epicatechin, a potent antioxidant, displayed a 1750-fold greater abundance in wild soybeans than in cultivated soybeans. Multiple polyphenols in the catechin biosynthesis pathway were significantly higher in wild soybeans, including phlorizin, taxifolin, quercetin 3- O -galactoside, cyanidin 3- O -glucoside, (+)-catechin, (-)-epiafzelechin, catechin-glucoside, and three proanthocyanidins. They showed significant positive correlations with each other and antioxidant activities, indicating their cooperative contribution to the high antioxidant activities of wild soybeans. Additionally, natural acylation related to functional properties was characterized in a diverse range of polyphenols. Our study reveals the comprehensive reprogramming of polyphenolic antioxidants during domestication, providing valuable insights for metabolism-assisted fortification of crop nutrition.

Strand-specific RNA-seq is a powerful tool for the discovery of novel transcripts, annotation of genomes, and profiling of gene expression levels. Tn5 transposase has been successfully applied in massive-scale sequencing projects; in particular, Tn5 adaptor modification is used in epigenetics, genomic structure, and chromatin visualization. We developed a novel dU-adaptor-assembled Tn5-mediated strand-specific RNA-sequencing protocol and compared this method with the leading dUTP method in terms of experimental procedure and multiple quality metrics of the generated libraries. The results showed that the dU-Tn5 method is easy to operate and generates a strand-specific RNA-seq library of comparable quality considering library complexity, strand-specificity, evenness, and continuity of annotated transcript coverage. We also evaluated the performance of the dU-Tn5 method in identifying nitrogen-responsive protein-coding genes and long non-coding RNAs in soybean roots. The results indicated that ~62-70% of differentially expressed genes detected from conventional libraries were also detected in dU-Tn5 libraries, indicating good agreement of our method with the current standard; moreover, their fold-changes were highly correlated (R>0.9). Thus, our method provides a promising 'do-it-yourself' stranded RNA-seq procedure for gene expression profiling., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides (nt) that are not translated into known functional proteins. This broad definition covers a large collection of transcripts with diverse genomic origins, biogenesis, and modes of action. Thus, it is very important to choose appropriate research methodologies when investigating lncRNAs with biological significance. Multiple reviews to date have summarized the mechanisms of lncRNA biogenesis, their localization, their functions in gene regulation at multiple levels, and also their potential applications. However, little has been reviewed on the leading strategies for lncRNA research. Here, we generalize a basic and systemic mind map for lncRNA research and discuss the mechanisms and the application scenarios of 'up-to-date' techniques as applied to molecular function studies of lncRNAs. Taking advantage of documented lncRNA research paradigms as examples, we aim to provide an overview of the developing techniques for elucidating lncRNA interactions with genomic DNA, proteins, and other RNAs. In the end, we propose the future direction and potential technological challenges of lncRNA studies, focusing on techniques and applications.

Chromium (Cr) toxicity impairs the productivity of crops and is a major threat to food security worldwide. However, the effect of Cr toxicity on seed germination and transcriptome of germinating seedlings of soybean crop has not been fully explored. In this study, two Cr-tolerant lines (J82, S125) and two Cr-sensitive ones (LD1, RL) were screened out of twenty-one soybean (Glycine max L.) genotypes based on seed germination rate, seed germinative energy, seed germination index, and growth of germinating seedlings under 50 mg L -1 Cr treatment. We found that Cr stress inhibits the growth of soybean seed germinating seedlings due to the Cr-induced overaccumulation of reactive oxygen species (ROS). Significantly different levels of element contents, antioxidant enzyme activities, malondialdehyde content were observed in the four soybean genotypes with contrasting Cr tolerance. Further, a total of 13,777 differentially expressed genes (DEGs) were identified in transcriptomic sequencing and 1298 DEGs in six gene modules were found highly correlated with the physiological traits by weighted correlation network analysis (WGCNA) analysis. The DEGs encoding antioxidant enzymes, transcription factors, and ion transporters are proposed to confer Cr tolerance in soybean germinating seedlings by reducing the uptake and translocation of Cr, decreasing the level of ROS, and keeping the osmotic balance in soybean germinating seedings. In conclusion, our study provided a molecular regulation network on soybean Cr tolerance at seed germinating stage and identified candidate genes for molecular breeding of low Cr accumulation soybean cultivars., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The considerable drought tolerance of wild cereal crop progenitors has diminished during domestication in the pursuit of higher productivity. Regaining this trait in cereal crops is essential for global food security but requires novel genetic insight. Here, we assessed the molecular evidence for natural variation of drought tolerance in wild barley (Hordeum spontaneum), wild emmer wheat (Triticum dicoccoides), and Brachypodium species collected from dry and moist habitats at Evolution Canyon, Israel (ECI). We report that prevailing moist vs dry conditions have differentially shaped the stomatal and photosynthetic traits of these wild cereals in their respective habitats. We present the genomic and transcriptomic evidence accounting for differences, including co-expression gene modules, correlated with physiological traits, and selective sweeps, driven by the xeric site conditions on the African Slope (AS) at ECI. Co-expression gene module 'circadian rhythm' was linked to significant drought-induced delay in flowering time in Brachypodium stacei genotypes. African Slope-specific differentially expressed genes are important in barley drought tolerance, verified by silencing Disease-Related Nonspecific Lipid Transfer 1 (DRN1), Nonphotochemical Quenching 4 (NPQ4), and Brassinosteroid-Responsive Ring-H1 (BRH1). Our results provide new genetic information for the breeding of resilient wheat and barley in a changing global climate with increasingly frequent drought events., (© 2022 The Authors New Phytologist © 2022 New Phytologist Foundation.)

(1) Background: Senescence represents the final stage of plant growth and development, which transfers nutrients to growing seeds and directly affects the yield and quality of crops. However, little is known about chlorophyll degradation in developing and maturing seeds, in contrast to leaf senescence; (2) Methods: RNA-Seq was used to analyze the differentially expressed genes of different late-senescent germplasms. A widely untargeted metabolic analysis was used to analyze differential metabolites. In addition, qRT-PCR was conducted to detect gene expression levels; (3) Results: Transcriptome analysis revealed that ZX12 seeds have a higher expression level of the chlorophyll synthesis genes in the early stage of maturity, compared with ZX4, and have a lower expression level of chlorophyll degradation genes in the late stage of maturity. Flavonoids were the primary differential metabolites, and ZX12 contains the unique and highest expression of three types of metabolites, including farrerol-7-O-glucoside, cyanidin-3-o-(6'-o-feruloyl) glucoside, and kaempferide-3-o-(6'-malonyl) glucoside. Among them, farrerol-7-O-glucoside and cyanidin-3-o-(6'-o-feruloyl) glucoside are flavonoid derivatives containing mono and dihydroxy-B-ring chemical structures, respectively; and (4) Conclusions: It is speculated that the two metabolites can slow down the degradation process of chlorophyll by scavenging oxygen-free radicals in the chloroplast.

Background: This study assessed the predictive value of uric acid (UA) for contrast-induced acute kidney injury (CI-AKI) in patients with type 2 diabetes mellitus (T2DM) who underwent coronary angiography (CAG). A nomogram to aid in the prediction of CI-AKI was also developed and validated, and the construction of a prognostic nomogram combined with clinical features was attempted., Methods: This study retrospectively enrolled T2DM patients who underwent CAG between December 2019 and December 2020 at the Affiliated Zhongda Hospital of Southeast University. Multivariable logistic regression analysis was used for the analysis of clinical outcomes. Receiver operating characteristic (ROC) analyses were performed to determine the area under the ROC curve (AUC) and the cut-off points for continuous clinical data. The prediction accuracies of models for CI-AKI were estimated through Harrell's concordance indices (C-index). Nomograms of the prognostic models were plotted for individualized evaluations of CI-AKI in T2DM patients after CAG., Results: A total of 542 patients with T2DM who underwent CAG were included in this study. We found that a high UA level (≥ 425.5 µmol/L; OR = 6.303), BUN level (≥ 5.98 mmol/L; OR = 3.633), Scr level (≥ 88.5 µmol/L; OR = 2.926) and HbA1C level (≥ 7.05%; OR = 5.509) were independent factors for CI-AKI in T2DM patients after CAG. The nomogram model based on UA, BUN, Scr and HbA1C levels presented outstanding performance for CI-AKI prediction (C-index: 0.878). Decision curve analysis (DCA) showed good clinical applicability in predicting the incidence of CI-AKI in T2DM patients who underwent CAG., Conclusion: High UA levels are associated with an increased incidence of CI-AKI in T2DM patients after CAG. The developed nomogram model has potential predictive value for CI-AKI and might serve as an economic and efficient prognostic tool in clinical practice., (© 2022. The Author(s).)

Background: Sarcopenia is prevalent and is associated with the occurrence of cardiovascular complications in patients undergoing maintenance hemodialysis (MHD). It is unknown how skeletal muscle may be associated with aspects of myocardial structure and function. This study aimed to evaluate the association between sarcopenia and cardiac structure and function in patients undergoing MHD. We also examined the prognostic role of sarcopenia for mortality and cardiovascular events (CVE) in this population., Methods: Participants from a single center underwent bioimpedance body composition analysis to measure skeletal muscle and echocardiography to assess myocardial structure and function. Sarcopenia was diagnosed based on the Asian Working Group for Sarcopenia criteria. The end points were all-cause mortality and CVE., Results: Of the 158 participants, 46 (29.1%) had sarcopenia, 102 (64.6%) had left ventricular diastolic dysfunction (LVDD), and 106 (67.0%) had left ventricular hypertrophy (LVH). Participants with sarcopenia had smaller right ventricular sizes (2.54 ± 0.77 vs 2.76 ± 0.28; P < 0.01), inter-ventricular thickness (1.07 ± 0.19 vs 1.14 ± 0.20; P = 0.039), and left ventricular posterior wall thickness (0.96, 0.89-1.10 vs 1.06, 0.95-1.20; P = 0.018). Skeletal muscle mass was strongly correlated with left ventricular mass (LVM) (r = 0.577; P < 0.0001). Furthermore, the risk of LVDD (OR: 4.92, 95% confidence interval (CI) [1.73-13.95]) and LVH (OR: 4.88, 95% CI [1.08-21.96]) was much higher in the sarcopenic group than in the non-sarcopenic group. During a follow-up period of 18 months, 11 (6.9%) patients died, of which seven died (4.4%) of CVE, and 36 (22.8%) experienced CVE. The presence of sarcopenia (adjusted hazard ratio (HR), 6.59; 95% CI [1.08-39.91]; P = 0.041) and low skeletal muscle index (HR, 3.41; 95% CI [1.01-11.57]; P = 0.049) and handgrip strength (HR, 0.88; 95% CI [0.78-0.99]; P = 0.037) independently predicted death. Sarcopenia was a significant predictor of CVE (HR, 10.96; 95% CI [1.14-105.10]; P = 0.038)., Conclusion: Our findings demonstrated that sarcopenia is associated with LVDD and LVH, and is associated with a higher probability of death and CVE., Competing Interests: The authors declare that they have no competing interests., (© 2022 Zhang et al.)

Purpose: This study aimed to clarify the formation and fixation of the annulus of Zinn (AZ) and its relationship with the extraocular muscles by using ultrathin plastination and three-dimensional models., Methods: Eighteen cadaveric heads (36 sides of the orbital apex) were plastinated to coronal (16 sides), sagittal (13 sides), and horizontal (5 sides) ultrathin plastination sections to be investigated at both macroscopic and microscopic levels. One cadaveric head was used for endoscopic dissection to identify anatomic landmarks., Results: There were two fibrous triangles adhered to both ends of the anterior surface of the optic strut. The superior rectus muscle originated from the superior fibrous triangle, and the lateral, inferior, and medial rectus muscles emerged from the inferior fibrous triangle. It was not until 5.46 ± 0.41 mm anterior to the optic strut that the complete tendinous ring composed of rectus muscles, optic nerve sheath, and periosteum was formed. The superior oblique and levator palpebrae superioris muscles originated from the medial fibrous band of the AZ. At the posterior of the AZ, there was a potential passage between the medial rectus muscle and the optic nerve., Conclusions: The fixation of the AZ was composed of the connection of the annular tendon to the optic strut posteriorly and the attachment of the complete tendinous ring to the lesser and greater wings of the sphenoid bone anteriorly. The triangular route area between the optic nerve and medial rectus muscle was located on the anterior side of the base of the optic strut.

The medial wall of the cavernous sinus (CS) has a significant role in evaluation and treatment of pituitary adenomas. This study was conducted to clarify the fine architecture of the medial wall and medial compartment of the CS at both macro- and micro-levels in twenty-one human cadaveric heads by using the epoxy sheet plastination technique. The sellar part medial wall is an intact dural layer that separates the CS from the pituitary gland. This dural wall adhered to the diaphragma sellae and the periosteum of the sella turcica to form fibrous triangles. Eight micro-protrusions of the pituitary gland were found at both sides of that wall. The thickness of the sellar part medial wall at its central portion was significantly thinner than that at the other surrounding portions. From the superior view, tortuous intracavernous carotid arteries can be divided into outward bending type and inward bending type. The inward bending intracavernous carotid was apt to bent towards the central part of the sellar part medial wall, where there were usually wide and short fibrous bands with more densely stained connective tissues between them. The micro-protrusion of the pituitary gland in the medial wall of the CS could provide an anatomical basis for the occult tumor invasion and the recurrence of residual tumor. Different bending facing states of tortuous intracavernous carotid arteries in the lateral direction may be a factor of the determination of the direction of growth of pituitary tumors., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The large size and complexity of most fern genomes have hampered efforts to elucidate fundamental aspects of fern biology and land plant evolution through genome-enabled research. Here we present a chromosomal genome assembly and associated methylome, transcriptome and metabolome analyses for the model fern species Ceratopteris richardii. The assembly reveals a history of remarkably dynamic genome evolution including rapid changes in genome content and structure following the most recent whole-genome duplication approximately 60 million years ago. These changes include massive gene loss, rampant tandem duplications and multiple horizontal gene transfers from bacteria, contributing to the diversification of defence-related gene families. The insertion of transposable elements into introns has led to the large size of the Ceratopteris genome and to exceptionally long genes relative to other plants. Gene family analyses indicate that genes directing seed development were co-opted from those controlling the development of fern sporangia, providing insights into seed plant evolution. Our findings and annotated genome assembly extend the utility of Ceratopteris as a model for investigating and teaching plant biology., (© 2022. The Author(s).)

P 2 -type Ca 2+ ATPases are responsible for cellular Ca 2+ transport, which plays an important role in plant development and tolerance to biotic and abiotic stresses. However, the role of P 2 -type Ca 2+ ATPases in stress response and stomatal regulation is still elusive in soybean. In this study, a total of 12 P 2 -type Ca 2+ ATPases genes ( GmACAs and GmECAs ) were identified from the genome of Glycine max . We analyzed the evolutionary relationship, conserved motif, functional domain, gene structure and location, and promoter elements of the family. Chlorophyll fluorescence imaging analysis showed that vegetable soybean leaves are damaged to different extents under salt, drought, cold, and shade stresses. Real-time quantitative PCR (RT-qPCR) analysis demonstrated that most of the GmACAs and GmECAs are up-regulated after drought, cold, and NaCl treatment, but are down-regulated after shading stress. Microscopic observation showed that different stresses caused significant stomatal closure. Spatial location and temporal expression analysis suggested that GmACA8, GmACA9, GmACA10, GmACA12, GmACA13 , and GmACA11 might promote stomatal closure under drought, cold, and salt stress. GmECA1 might regulate stomatal closure in shading stress. GmACA1 and GmECA3 might have a negative function on cold stress. The results laid an important foundation for further study on the function of P 2 -type Ca 2+ ATPase genes GmACAs and GmECAs for breeding abiotic stress-tolerant vegetable soybean., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Fu, Zhang, Chen, Li, Shangguan, Zheng, Xu, Chen and Xu.)

Formalin-fixed, paraffin-embedded (FFPE) tissues have been widely used in researches. Proteins and nucleic acids in prolonged FFPE tissues display different degrees of degradation. We investigated the effect of prolonged formalin fixation on protein expression in human brain tissues. Twenty-eight middle prefrontal front cortex tissue blocks from human brains prefixed in formalin were obtained from a brain bank. The tissue blocks were divided into two groups, the control group and the prolonged fixation group. Quantitative immunocytochemistry was used to analyse the biological markers of Fox-3, Rbfox3 (NeuN), glial fibrillary acidic protein (GFAP), ionized calcium binding adapter molecule-1 (IBA-1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Nissl staining showed that positive signaling of Nissl body was significantly decreased by 16.6% in the prolonged fixation group. In addition, the staining intensity of Nissl body was negatively correlated with fixation time. The level of NeuN immunoreactivity (ir) was significantly reduced by 19.31% in the prolonged fixation group. Moreover, there was a significant negative correlation between NeuN-ir and fixation time. There were no significant changes in GFAP-ir, IBA-1-ir and GAPDH-ir between control group and the prolonged fixation group. Prolonged formalin-fixed tissues showed time- and molecule-dependent protein changes, which may be potential confounders in the clinic and researches. Our study suggested short formalin fixation time is recommended when using PPFE brain tissues., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Tobacco (Nicotiana tabacum) is one of the most widely cultivated commercial non-food crops with significant social and economic impacts. Here we profiled transcriptome and metabolome from 54 tobacco samples (2-3 replicates; n = 151 in total) collected from three varieties (i.e. genetic factor), three locations (i.e. environmental factor), and six developmental stages (i.e. developmental process). We identified 3,405 differentially expressed (DE) genes (DEGs) and 371 DE metabolites, respectively. We used quantitative real-time PCR to validate 20 DEGs, and confirmed 18/20 (90%) DEGs between three locations and 16/20 (80%) with the same trend across developmental stages. We then constructed nine co-expression gene modules and four co-expression metabolite modules , and defined seven de novo regulatory networks, including nicotine- and carotenoid-related regulatory networks. A novel two-way Pearson correlation approach was further proposed to integrate co-expression gene and metabolite modules to identify joint gene-metabolite relations. Finally, we further integrated DE and network results to prioritize genes by its functional importance and identified a top-ranked novel gene, LOC107773232, as a potential regulator involved in the carotenoid metabolism pathway. Thus, the results and systems-biology approaches provide a new avenue to understand the molecular mechanisms underlying complex genetic and environmental perturbations in tobacco., (© The Author(s) 2020. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.)

Objective: Kambin's triangle and the safe triangle are common posterolateral approaches for lumbar transforaminal endoscopic surgery and epidural injection. To date, no consensus has been reached on the optimal transforaminal approach, in particular its underlying anatomical mechanism. The aim of this study was to investigate the 3D architecture of the neurovascular and adipose zones in the upper and lower lumbar intervertebral foramina (IVFs)., Methods: Using the epoxy sheet plastination technology, 22 cadaveric lumbar spines (12 female and 10 male, age range 46-89 years) were prepared as a series of transverse (11 sets), sagittal (8 sets), and coronal (3 sets) slices with a thickness of 0.25 mm (6 sets) or 2.5 mm (16 sets). The high-resolution images of the slices were scanned and analyzed. The height, area, and volume of 30 IVFs from T12-L1 to L4-5 were estimated and compared. This study was performed in accord with the authors' institutional ethical guidelines and approved by the institutional ethics committees., Results: The findings were as follows. 1) The 3D boundaries of the lumbar IVF and its subdivisions were precisely defined. 2) The 3D configuration of the neurovascular and adipose zones was different between the upper and lower lumbar IVFs; zoning in the upper lumbar IVFs was much more complex than that in the lower lumbar IVFs. 3) In general, the infraneural adipose zone gradually tapered and rotated from the inferoposterolateral aspect to the superoanteromedial aspect. 4) The average height, area, and volume of the IVF gradually increased from the upper to the lower lumbar spine. Within a lumbar IVF, the volumes below and above the inferior border of the dorsal root ganglia were similar., Conclusions: This study highlights differences of fine 3D architecture of neurovascular and adipose tissues between the upper and lower lumbar IVFs, with related effects on the transforaminal approaches. The findings may contribute to optimization of the surgical approaches to and through the IVF at different lumbar spinal levels and also may help to shorten the learning curve for the transforminal techniques.

Objective: This 3-dimensional histologic study aimed to provide a precise description of the meningeal structures in the jugular foramen., Methods: 22 posterior skull base tissue blocks containing the jugular foramen region were obtained from 11 human cadaveric heads. These blocks were plastinated and cut into serial sections. After staining, these sections were examined under an optical microscope and used to reconstruct a 3-dimensional visualization model., Results: At the intracranial orifice of the jugular foramen, the meningeal dura formed 2 separate dural perforations: the glossopharyngeal meatus and the vagal meatus. The arachnoid extended into 2 dural meatuses and terminated at the inferior ganglion of the glossopharyngeal nerve in the glossopharyngeal meatus and the superior ganglion of the vagus nerve in the vagal meatus. At the intraforaminal part of the jugular foramen, the meningeal dura encased the glossopharyngeal nerve to form a dural sheath while encasing the vagus and accessory nerves to form a dural network. At the extracranial orifice of the jugular foramen, the dural wall of the jugular bulb extended downward to form a dense connective tissue sheath. The initial end of the internal jugular vein invaginated into this sheath and fused with the jugular bulb., Conclusions: Knowledge of the anatomy of the meningeal architecture of the jugular foramen can be helpful in avoiding surgical complications of the lower cranial nerves when this complex area is approached., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Histone H2B protein is not only structurally important for chromosomal DNA packaging but is also involved in the regulation of gene expression, including the immune response of plants against pathogens. In this study, we show that the potato virus X (PVX) infection resulted in the reduced expression of H2B at both the mRNA and protein level in Nicotiana benthamiana . Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) was then used to down-regulate the expression of H2B in N. benthamiana and tests showed that the titre of TRV was similar in these plants to that in control treated plants. When these H2B-silenced plants were inoculated with PVX, the virus spread more slowly through the plant and there was a lower titre of PVX compared to non-silenced plants. Abnormal leaf development and stem necrosis were observed in the H2B-silenced plants, which were alleviated in H2B-silenced NahG transgenic plants suggesting the involvement of salicylic acid (SA) in the production of these symptoms. Indeed, quantitative reverse transcription (qRT)-PCR and liquid chromatography tandem mass spectroscopy (LC-MS) results showed that endogenous SA is increased in H2B-silenced N. benthamiana . Thus, downregulation of H2B induced the accumulation of endogenous SA, which was correlated with stem necrosis and a decreased accumulation of PVX in N. benthamiana .

Background: Extracellular accumulation of amyloid β-peptide (Aβ) is one of pathological hallmarks of Alzheimer's disease (AD) and contributes to the neuronal loss. Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an endoplasmic reticulum (ER) stress-inducible neurotrophic factor. Many groups, including ours, have proved that MANF rescues neuronal loss in several neurological disorders, such as Parkinson's disease and cerebral ischemia. However, whether MANF exerts its protective effect against Aβ neurotoxicity in AD remains unknown., Methods: In the present study, the characteristic expressions of MANF in Aβ 1-42 -treated neuronal cells as well as in the brains of APP/PS1 transgenic mice were analyzed by immunofluorescence staining, qPCR, and Western blot. The effects of MANF overexpression, MANF knockdown, or recombination human MANF protein (rhMANF) on neuron viability, apoptosis, and the expression of ER stress-related proteins following Aβ 1-42 exposure were also investigated., Results: The results showed the increased expressions of MANF, as well as ER stress markers immunoglobulin-binding protein (BiP) and C/EBP homologous protein (CHOP), in the brains of the APP/PS1 transgenic mice and Aβ 1-42 -treated neuronal cells. MANF overexpression or rhMANF treatment partially protected against Aβ 1-42 -induced neuronal cell death, associated with marked decrease of cleaved caspase-3, whereas MANF knockdown with siRNA aggravated Aβ 1-42 cytotoxicity including caspase-3 activation. Further study demonstrated that the expressions of BiP, ATF6, phosphorylated-IRE1, XBP1s, phosphorylated-eIF2α, ATF4, and CHOP were significantly downregulated by MANF overexpression or rhMANF treatment in neuronal cells following Aβ 1-42 exposure, whereas knockdown of MANF has the opposite effect., Conclusions: These findings demonstrate that MANF may exert neuroprotective effects against Aβ-induced neurotoxicity through attenuating ER stress, suggesting that an applicability of MANF as a therapeutic candidate for AD.

Leaf shape is an important trait that influences the utilization rate of light, and affects quality and yield of pea ( Pisum sativum ). In the present study, a joint method of high-density genetic mapping using specific locus amplified fragment sequencing (SLAF-seq) and bulked segregant analysis (BSA) was applied to rapidly detect loci with leaf shape traits. A total of 7,146 polymorphic SLAFs containing 12,213 SNP markers were employed to construct a high-density genetic map for pea. We conducted quantitative trait locus (QTL) mapping on an F 2 population to identify QTLs associated with leaf shape traits. Moreover, SLAF-BSA was conducted on the same F 2 population to identify the single nucleotide polymorphism (SNP) markers linked to leaf shape in pea. Two QTLs (qLeaf&#95;or-1, qLeaf&#95;or-2) were mapped on linkage group 7 (LG7) for pea leaf shape. Through alignment of SLAF markers with Cicer arietinum, Medicago truncatula , and Glycine max , the pea LGs were assigned to their corresponding homologous chromosomal groups. The comparative genetic analysis showed that pea is more closely related to M. truncatula . Based on the sequencing results of two pools with different leaf shape, 179 associated markers were obtained after association analysis. The joint analysis of SLAF-seq and BSA showed that the QTLs obtained from mapping on a high-density genetic map are convincing due to the closely associated map region with the BSA results, which provided more potential markers related to leaf shape. Thus, the identified QTLs could be used in marker-assisted selection for pea breeding in the future. Our study revealed that joint analysis of QTL mapping on a high-density genetic map and BSA-seq is a cost-effective and accurate method to reveal genetic architecture of target traits in plant species without a reference genome.

Although the link between stress and alcohol is well recognized, the underlying mechanisms of how they interplay at the molecular level remain unclear. The purpose of this study is to identify molecular networks underlying the effects of alcohol and stress responses, as well as their interaction on anxiety behaviors in the hippocampus of mice using a systems genetics approach. Here, we applied a gene co-expression network approach to transcriptomes of 41 BXD mouse strains under four conditions: stress, alcohol, stress-induced alcohol and control. The co-expression analysis identified 14 modules and characterized four expression patterns across the four conditions. The four expression patterns include up-regulation in no restraint stress and given an ethanol injection (NOE) but restoration in restraint stress followed by an ethanol injection (RSE; pattern 1), down-regulation in NOE but rescue in RSE (pattern 2), up-regulation in both restraint stress followed by a saline injection (RSS) and NOE, and further amplification in RSE (pattern 3), and up-regulation in RSS but reduction in both NOE and RSE (pattern 4). We further identified four functional subnetworks by superimposing protein-protein interactions (PPIs) to the 14 co-expression modules, including γ-aminobutyric acid receptor (GABA) signaling, glutamate signaling, neuropeptide signaling, cAMP-dependent signaling. We further performed module specificity analysis to identify modules that are specific to stress, alcohol, or stress-induced alcohol responses. Finally, we conducted causality analysis to link genetic variation to these identified modules, and anxiety behaviors after stress and alcohol treatments. This study underscores the importance of integrative analysis and offers new insights into the molecular networks underlying stress and alcohol responses.

17 β -Estradiol (E2) is a multifunctional steroid hormone in modulating metabolism in vivo . Previous studies have reported that E2 could promote insulin secretion and protect β cells from apoptosis. In this study, the partial pancreatectomy (PPx) model was used to study the role of E2 in islet cell proliferation. The animals were divided into four groups, including sham control, PPx model, E2, and E2 plus estrogen antagonist (E2 plus ICI) groups. In the E2 group, 5-bromo-2'-deoxyuridine- and Ki67-positive cells significantly increased after PPx, and the protein expression of forkhead transcription factor M1, cyclin A2, cyclin B1, and cyclin E2 also significantly increased in the isolated islets. The messenger RNA expression of cyclin A2 and cyclin B2 increased in E2 treatment group. Additionally, the effects of E2 on the PPx mice were partially blocked by estrogen antagonist ICI182,780. The results indicated that E2 significantly promoted islet cell proliferation in PPx model mice, and it upregulated the expression of cell cycle genes. In conclusion, E2 treatment is beneficial for islet cell proliferation in adult mice after PPx. A partial pancreatectomy in mice may be an attractive model for the study of islet cell proliferation.

Chilling stress is a major factor limiting the yield and quality of vegetable soybean (Glycine max L.) on a global scale. In the present study, systematic identification and functional analysis of miRNAs under chilling stress were carried out to clarify the molecular mechanism of chilling resistance. Two independent small RNA libraries from leaves of soybean were constructed and sequenced with the high-throughput Illumina Solexa system. A total of 434 known miRNAs and 3 novel miRNAs were identified. Thirty-five miRNAs were verified by qRT-PCR analysis. Furthermore, their gene targets were identified via high-throughput degradome sequencing. A total of 898 transcripts were targeted by 54 miRNA families attributed to five categories. More importantly, we identified 51 miRNAs differentially expressed between chilling stress and control conditions. The targets of these miRNAs were enriched in oxidation-reduction, signal transduction, and metabolic process functional categories. Our qRT-PCR analysis confirmed a negative relationship among the miRNAs and their targets under chilling stress. Our work thus provides comprehensive molecular evidence supporting the involvement of miRNAs in chilling-stress responses in vegetable soybean.

Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area.

Understanding the molecular mechanisms regulating pea seed developmental process is extremely important for pea breeding. In this study, we used high-throughput RNA-Seq and bioinformatics analyses to examine the changes in gene expression during seed development in vegetable pea and grain pea, and compare the gene expression profiles of these two pea types. RNA-Seq generated 18.7 G of raw data, which were then de novo assembled into 77,273 unigenes with a mean length of 930 bp. Our results illustrate that transcriptional control during pea seed development is a highly coordinated process. There were 459 and 801 genes differentially expressed at early and late seed maturation stages between vegetable pea and grain pea, respectively. Soluble sugar and starch metabolism related genes were significantly activated during the development of pea seeds coinciding with the onset of accumulation of sugar and starch in the seeds. A comparative analysis of genes involved in sugar and starch biosynthesis in vegetable pea (high seed soluble sugar and low starch) and grain pea (high seed starch and low soluble sugar) revealed that differential expression of related genes at late development stages results in a negative correlation between soluble sugar and starch biosynthetic flux in vegetable and grain pea seeds. RNA-Seq data was validated by using real-time quantitative RT-PCR analysis for 30 randomly selected genes. To our knowledge, this work represents the first report of seed development transcriptomics in pea. The obtained results provide a foundation to support future efforts to unravel the underlying mechanisms that control the developmental biology of pea seeds, and serve as a valuable resource for improving pea breeding.

Abuse and dependence to heroin has evolved into a global epidemic as a significant clinical and societal problem with devastating consequences. Repeated exposure to heroin can induce long-lasting behavioral sensitization and withdrawal. Pharmacological activation of 5-HT2C receptors (5-HT2CRs) suppresses psychostimulant-induced drug-seeking and behavioral sensitization. The present study examined the effect of a selective 5-HT2CR agonist lorcaserin on behavioral sensitization and naloxone-precipitated withdrawal symptoms in heroin-treated mice. Male mice received heroin (1.0 mg/kg, s.c.) twice a day for 3 days and then drug treatment was suspended for 5 days. On day 9, a challenge dose of heroin (1.0 mg/kg) was administered to examine the expression of behavioral sensitization. Lorcaserin administered during the development, withdrawal or expression stage suppressed heroin-induced behavioral sensitization on day 9. Another cohort of mice received increasing doses of heroin over a 4.5-day period. Lorcaserin, or the positive control clonidine (an α2-adrenoceptor agonist) suppressed naloxone-precipitated withdrawal symptoms in heroin-treated mice. These findings suggest that activation of 5-HT2CRs suppresses behavioral sensitization and withdrawal in heroin-treated mice. Thus, pharmacological activation of 5-HT2CRs may represent a new avenue for the treatment of heroin addiction., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Mesencephalic astrocyte-derived neurotrophic factor (MANF) has been shown to be up-regulated under the focal cerebral ischemia and protected against ischemic injury in rats. However, the underlying mechanisms are unclear. The aim of this study was to verify the protection of MANF on the cerebral ischemic injury and further investigate the possible mechanisms. Rat focal ischemic model was established by middle cerebral artery occlusion (MCAO). The recombinant human MANF was therapeutically administrated to the ipsilateral ventricle at 2 h after MCAO. MANF decreased the number of the propidium iodide (PI)- and TUNEL-positive neural cells. Contrarily, MANF protected the NeuN-positive cells in hippocampus and cortex from death induced by ischemia. The more interesting results in this study were that MANF repressed the cleavage of caspase-3 triggered by focal cerebral ischemia. MANF also reduced the elevated levels of BIP/Grp78, phosphorylated IRE1, and splicing XBP1 induced by focal cerebral ischemia, but not affect CHOP expression. Meanwhile, focal cerebral ischemia elevated the levels of XBP1 mRNA, including unspliced XBP1 (XBP1u) and spliced XBP1 (XBP1s). However, MANF did not affect the expression of XBP1 mRNA, neither XBP1u nor XBP1s. These results suggest that MANF can prevent the neuron loss via inhibiting ischemia-induced apoptosis and regulating unfolded protein response-related genes., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: No consensus has been obtained on the differences between class IV-S and IV-G lupus nephritis (LN), especially regarding renal outcome. Our study investigated clinical-pathological features and prognosis of diffuse segmental and pure diffuse global proliferative LN., Methods: In this retrospective study, a total of 120 patients with biopsy-proven diffuse LN were included, of which 31 patients were class IV-S and 89 were pure class IVG. Class IV-S was defined as segmental lesion involving ≥ 50% of all glomeruli, while pure class IV-G was defined as global lesion involving ≥ 50% of all glomeruli with no segmental necrosis or crescents. The clinical- pathological and prognostic features of the two classes were compared., Results: There was no difference in levels of urine protein or serum creatinine between the two groups. Higher serological activity was observed in the pure IV-G group with lower complement C3 (p < 0.001) and C4 level (p < 0.001), compared to the IV-S group. Histologically, immune-complex deposits were significantly more common in the pure IV-G group, with higher prevalence of wire loop (42.7% vs. 0%, p < 0.001) and hyaline thrombi (34.8% vs. 3.2%, p < 0.001). However, the complete remission (CR) rate to intravenous cyclophosphamide (IVCY) induction was lower in the IV-S than in the pure IV-G group (16.7% vs. 53.2%, p = 0.023). After 1 year, the pure class IV-G group had a higher CR rate (71.9% vs. 48.4%, p = 0.017). The 10-year renal survival rate (without doubling of serum creatinine or end-stage renal disease) was significantly lower in patients with IV-S than pure IV-G (75.2% vs. 97.4%, p = 0.028)., Conclusion: LN class IV-S and class IV-G without segmental lesion showed different clinical-histological features and prognosis, suggesting that different mechanisms may exist.

Objective: To provide the anatomic data for operation on the middle ear through the observation and measurement of related anatomic structure., Method: Forty human temporal bones of 20 voluntary bone donors were dissected, relative anatomical data of operation were observed and measured under operating microscope through posterior tympanum approach entering posterior tympanum., Result: The average distances from suprameatal spine to short crus of incus, pyramid segment of facial canal were 19.14 mm, 16.30 mm, respectively. The average distances from pyramid segment of facial canal to the surface of mastoid, crotch of chorda tympani nerve, posterior wall of auditory meatus were 20.84 mm, 11.28 mm, 4.40 mm, respectively. The average length of facial nerve in the horizontal segment, the perpendicular paragraph was about 11.60 mm, 15.30 mm, respectively. The average distance from pyramidal eminence to the anterior lip of round window niche, from oval window to round window niche, from incudostapedial joint to round window niche was 4.46 mm, 3.74 mm, 3.80 mm, respectively. The included angle of facial nerve in the horizontal segment and chorda tympani nerve with facial nerve in the perpendicular paragraph was 110.4 degrees, 24.8 degrees, respectively. Horizontal semicircular canal and facial nerve in the level paragraph was 17.5 degrees, long process of incus and incus buttress was 46.0 degrees., Conclusion: The position of anatomic structure in middle ear was constant and the relationship including distance and angle between anatomic structures changed in limited region. The anatomical parameters provide a reference value for avoiding the injury during the operation.

Objective: To provide the clinical anatomy data in physiological condition of the lingual artery in 64-slices spiral computed tomography angiography (CTA) for clinical treatment concerning the lingual artery., Method: CTA examination of the carotid artery were performed on 80 adult volunteers. The 3D reconstruction images of the carotid artery with hyoid bone were obtained by using 64-slices spiral CT and image postprocessing workstation. The origin, pathway, and anatomic relations of the lingual artery in CTA were studied. The distance from the origin of the lingual artery to the bifurcation of the common carotid artery and tip of the greater horn of hyoid bone were separately measured, and the distance between the segment of the greater horn of hyoid bone of lingual artery and the middle of greater horn of hyoid bone were also measured., Result: The pathway of the lingual artery had high variation, but the relative position between the segment of the greater horn of hyoid bone of lingual artery and the greater horn of hyoid bone were relatively constant. The lingual artery run forward approximately parallel to the greater horn of hyoid bone into tongue (2.32 +/- 1.29) mm in the superior to the greater horn of hyoid bone or (2.00 +/- 1.68) mm in the inferior to the greater horn of hyoid bone The distances from the origin of the lingual artery to the bifurcation of the common carotid artery and tip of the greater horn of hyoid bone were (12.93 +/- 7.36) mm and (10.40 +/- 5.75) mm separately., Conclusion: The lingual artery could be clearly shown in CTA and the anatomy data in physiological condition of the lingual artery could be obtained by CTA. There was important instructive significance for clinical treatment concerning about the lingual artery.

Objectives: The pharyngeal and esophageal shortenings during deglutition are attributable to longitudinal muscle contraction. The two shortenings may be independent events because the longitudinal pharyngeal and esophageal muscular insertions are believed to be separated from each other. The aim of this study was to investigate the superior attachments of the longitudinal esophageal muscle., Study Design: Prospective., Methods: With use of dissection, sectional anatomy, and confocal microscopy, 30 adult cadavers were examined in this study., Results: We found that the longitudinal esophageal muscle fibers had multiple superior insertions. Some of the fibers were continuous with the longitudinal pharyngeal muscles and merged with a membrane-like fascia that anchored the pharyngoesophageal wall to the thyroid cartilage., Conclusions: This study demonstrates that the longitudinal pharyngeal and esophageal muscle fibers are continuous with each other and are integrated with fibrous and cartilaginous structures. Such anatomic integrity at the pharyngoesophageal junction is important for the function of the upper esophageal sphincter.

Cricopharyngeal bars are commonly seen on a barium swallow radiologic examination and represent the failure of the cricopharyngeus to relax. Traditionally, the bars have been considered as functional or physiologic protrusions. Recently, anatomical cricopharyngeal protrusion has been found in about 30% of cadavers of the elderly, suggesting that such a structural change may become a physical barrier that affects the normal deglutition in a living person. This suggests that such a radiographic finding should be carefully considered when interpreting radiologic and manometrical examinations and managing dysphagia of the elderly. However, the finding of the anatomical cricopharyngeal protrusion was based on the observation of cadavers of the elderly (mean age = 77 years). The aim of this study was to further investigate whether such an anatomical cricopharyngeal protrusion exists in cadavers of other ages. Using the dissection method, we examined 63 human cadavers that were divided into three age groups: young adult (6 females and 10 males, age = 16-24 years old), adult (8 females and 23 males, age = 25-64 years old), and early elderly (5 females and 11 males, age = 65-69 years old). We found that 57 of the 63 cadavers had a smooth mucosal surface on the posterior hypopharyngeal and upper esophageal wall. A slightly thickened posterior muscular wall was observed at the cricoid level in six cadavers (five from the adult group and one from the early elderly group), but its boundary was not clearly identified. Taking the previous reports together, our results suggest that an anatomical cricopharyngeal protrusion is closely associated with the aging process.