Marker Lines Research Articles

The Glycine-Rich Domain Protein GRDP2 Regulates Ovule Development via the Auxin Pathway in Arabidopsis.

The glycine-rich domain proteins (GRDP) have been functionally implicated in the cell wall structure, biotic, and abiotic stress responses. However, little is known about GRDP genes in female gametophyte development of Arabidopsis. This study shows that GRDP2, a GRDP, plays a crucial role in female gametophyte development. In GRDP2 overexpression lines, grdp2-3, the embryo sacs were arrested at FG1 and no nucleus stages. Furthermore, callose staining shows that cell plate formation during megasporogenesis is disturbed in grdp2-3. In contrast, the pollen development is not affected in grdp2-3. The expression patterns of auxin-specific marker lines in female gametophytes showed that the auxin distribution and transport were significantly changed during megagametogenesis in grdp2-3. In addition, compared with the membrane-localized pattern of PIN1, PIN2, and PIN7 in WT, the signals were detected in the cytoplasm in grdp2-3. Together, our data suggest that GRDP2 plays an essential role in auxin-mediated female gametophyte development.

Spatiotemporal relationship between auxin dynamics and hydathode development in Arabidopsis leaf teeth

ABSTRACT Hydathode is a plant tissue of vascular plants involved in water release called guttation. Arabidopsis hydathodes are found at the tips of leaf teeth and contain three major components: water pores, xylem ends, and small cells. Leaf teeth are known as the main parts for auxin biosynthesis and accumulation during leaf development. However, the detailed spatiotemporal relationship between auxin dynamics and hydathode development is unknown. In this study, we show that auxin biosynthesis and accumulation precede hydathode development. A triple marker line (called YDE line) containing three leaf tooth markers: YUC4:nls-3xGFP (auxin biosynthesis), DR5rev:erRFP (auxin accumulation or maxima), and E325-GFP (hydathode development), was generated, and spatiotemporal confocal microscopic analysis was carried out. The expression area of these markers became larger during leaf development, implying that the hydathode size enlarges as the leaf tooth grows. Detailed observation revealed that the auxin-related markers YUC4:nls-GFP and DR5rev:erRFP were first expressed in the early stage of leaf tooth growth. Then, E325-GFP was expressed partly overlapping with the auxin markers at a later stage. These findings provide new insights into the spatiotemporal relationship between auxin dynamics and hydathode development in Arabidopsis.

Preliminary Study of CT Three-dimensional Reconstruction Combined with Ground Glass Nodules of Natural Lung Collapse in Thoracoscopic Pulmonary Segmental Resection

背景与目的计算机断层扫描（computed tomography, CT）三维重建技术越来越多地被应用于肺磨玻璃结节（ground glass nodule, GGN）肺癌患者术前规划中，但术中如何准确定位结节和保证安全切除边缘依然是临床医生面对的难题。本研究旨在探讨全胸腔镜下肺段切除术中，CT三维重建联合术中肺自然萎陷定位方法的准确性、便捷性及切缘的安全性。方法选取2019年7月-2019年12月收治入院的45例影像学表现有肺GGN的患者为研究组，45例患者均接受薄层CT扫描并术前进行三维重建，于麻醉后快速打开胸腔小操作口和患者气道，利用压强差使肺快速自然萎陷，根据自然标志线进行GGN定位，3-0 prolene线标记，然后行三维重建指导下的胸腔镜肺段切除术。标本摘除后测量GGN与缝线标识的距离、GGN与切缘的距离并常规送检切缘。统计患者一般临床资料、病理资料与术后并发症，并与同期采用hookwire定位针进行定位的连续45例患者进行比较。结果CT三维重建联合术中无创式肺自然萎陷GGN定位，平均定位时间为6.9 min，定位准确率为90.6%。术中发现广泛胸腔粘连2例，肺气肿1例。术后病理均证实为肺腺癌，切缘送检均呈阴性。肺段切除后均无严重并发症的发生。结论CT三维重建联合术中肺自然萎陷GGN定位，缩短了术中寻找GGN的时间，保障了切缘的安全性，是一种更经济、实惠、便捷的定位方法，使肺段切除更加精准。

A molecular toolkit for the green seaweed Ulva mutabilis.

The green seaweed Ulva mutabilis is an ecologically important marine primary producer as well as a promising cash crop cultivated for multiple uses. Despite its importance, several molecular tools are still needed to better understand seaweed biology. Here, we report the development of a flexible and modular molecular cloning toolkit for the green seaweed U. mutabilis based on a Golden Gate cloning system. The toolkit presently contains 125 entry vectors, 26 destination vectors, and 107 functionally validated expression vectors. We demonstrate the importance of endogenous regulatory sequences for transgene expression and characterize three endogenous promoters suitable to drive transgene expression. We describe two vector architectures to express transgenes via two expression cassettes or a bicistronic approach. The majority of selected transformants (50%-80%) consistently give clear visual transgene expression. Furthermore, we made different marker lines for intracellular compartments after evaluating 13 transit peptides and 11 tagged endogenous Ulva genes. Our molecular toolkit enables the study of Ulva gain-of-function lines and paves the way for gene characterization and large-scale functional genomics studies in a green seaweed.

Iso-anchorene is an endogenous metabolite that inhibits primary root growth in Arabidopsis.

Carotenoid-derived regulatory metabolites and hormones are generally known to arise through the oxidative cleavage of a single double bond in the carotenoid backbone, which yields mono-carbonyl products called apocarotenoids. However, the extended conjugated double bond system of these pigments predestines them also to repeated cleavage forming dialdehyde products, diapocarotenoids, which have been less investigated due to their instability and low abundance. Recently, we reported on the short diapocarotenoid anchorene as an endogenous Arabidopsis metabolite and specific signaling molecule that promotes anchor root formation. In this work, we investigated the biological activity of a synthetic isomer of anchorene, iso-anchorene, which can be derived from repeated carotenoid cleavage. We show that iso-anchorene is a growth inhibitor that specifically inhibits primary root growth by reducing cell division rates in the root apical meristem. Using auxin efflux transporter marker lines, we also show that the effect of iso-anchorene on primary root growth involves the modulation of auxin homeostasis. Moreover, by using liquid chromatography-mass spectrometry analysis, we demonstrate that iso-anchorene is a natural Arabidopsis metabolite. Chemical inhibition of carotenoid biosynthesis led to a significant decrease in the iso-anchorene level, indicating that it originates from this metabolic pathway. Taken together, our results reveal a novel carotenoid-derived regulatory metabolite with a specific biological function that affects root growth, manifesting the biological importance of diapocarotenoids.

Utility of photodynamic diagnosis plus reflectance confocal microscopy in detecting the margins of extramammary Paget disease.

Due to the clinically poorly delineated unclear margin of extramammary Paget disease, the recurrence rate after surgical resection is high. To compare photodynamic diagnosis and photodynamic plus reflectance confocal microscopy diagnosis in determining the tumor margins in patients with extramammary Paget disease. Thirty-six patients with histopathologically confirmed primary extramammary Paget disease between January 2017 to June 2018 were included in the study. The skin lesion margins were preoperatively observed by the naked eye and with photodynamic diagnosis and photodynamic diagnosis plus reflectance confocal microscopy and they were compared to the postoperative histopathological examination results. Among the 130 sections taken from 36 patients, 83 sections (63.8%, 83/130) had tumor margins beyond the macroscopic line with a distance of 3.5 ± 3.1mm and a median of 2.7mm. Forty-six sections (35.4%, 46/130) exceeded the photodynamic diagnosis marker line with a distance of 2.1 ± 1.7mm and a median of 1.5mm. Twenty seven sections (20.8%, 27/130) were obtained beyond the photodynamic diagnosis plus reflectance confocal microscopy marker line with a distance of 1.4 ± 1.2mm and a median of 0.9mm. Photodynamic diagnosis and reflectance confocal microscopy detection can be used to observe only the superficial margin of the tumor and not the deep part. Moreover, reflectance confocal microscopy was not used alone as a control. In terms of determining the extramammary Paget disease margin invasively, photodynamic diagnosis and photodynamic diagnosis plus reflectance confocal microscopy were found superior to observations made with the naked eye, while photodynamic diagnosis plus reflectance confocal microscopy was superior to photodynamic diagnosis alone.

Volatile compounds from beneficial rhizobacteria Bacillus spp. promote periodic lateral root development in Arabidopsis.

Lateral root formation is coordinated by both endogenous and external factors. As biotic factors, plant growth-promoting rhizobacteria can affect lateral root formation, while the regulation mechanism is unclear. In this study, by applying various marker lines, we found that volatile compounds (VCs) from Bacillus amyloliquefaciens SQR9 induced higher frequency of DR5 oscillation and prebranch site formation, accelerated the development and emergence of the lateral root primordia and thus promoted lateral root development in Arabidopsis. We demonstrated a critical role of auxin on B. amyloliquefaciens VCs-induced lateral root formation via respective mutants and pharmacological experiments. Our results showed that auxin biosynthesis, polar transport and signalling pathway are involved in B. amyloliquefaciens VCs-induced lateral roots formation. We further showed that acetoin, a major component of B. amyloliquefaciens VCs, is less active in promoting root development compared to VC blends from B. amyloliquefaciens, indicating the presence of yet uncharacterized/unknown VCs might contribute to B. amyloliquefaciens effect on lateral root formation. In summary, our study revealed an auxin-dependent mechanism of B. amyloliquefaciens VCs in regulating lateral root branching in a non-contact manner, and further efforts will explore useful VCs to promote plant root development.

Development of the Middle Layer in the Anther of Arabidopsis.

The middle layer is an essential cell layer of the anther wall located between the endothecium and tapetum in Arabidopsis. Based on sectioning, the middle layer was found to be degraded at stage 7, which led to the separation of the tapetum from the anther wall. Here, we established techniques for live imaging of the anther. We created a marker line with fluorescent proteins expressed in all anther layers to study anther development. Several staining methods were used in the intact anthers to study anther cell morphology. We clarified the initiation, development, and degradation of the middle layer in Arabidopsis. This layer is initiated from both the inner and outer secondary parietal cells at stage 4, stopped cell division at stage 6, and finally degraded at stage 11. The neighboring cell layers, the epidermis, and endothecium continued cell division until stage 10, which led to a thin middle layer. The degradation of the tapetum cell wall at stage 7 lead to its isolation from the anther wall. This work presents fundamental information on the development of the middle layer, which facilitates the further investigation of anther development and plant fertility. These live imaging methods could be useful in future studies.

Reliable, scalable functional genetics in bloodstream-form Trypanosoma congolense in vitro and in vivo.

Animal African trypanosomiasis (AAT) is a severe, wasting disease of domestic livestock and diverse wildlife species. The disease in cattle kills millions of animals each year and inflicts a major economic cost on agriculture in sub-Saharan Africa. Cattle AAT is caused predominantly by the protozoan parasites Trypanosoma congolense and T. vivax, but laboratory research on the pathogenic stages of these organisms is severely inhibited by difficulties in making even minor genetic modifications. As a result, many of the important basic questions about the biology of these parasites cannot be addressed. Here we demonstrate that an in vitro culture of the T. congolense genomic reference strain can be modified directly in the bloodstream form reliably and at high efficiency. We describe a parental single marker line that expresses T. congolense-optimized T7 RNA polymerase and Tet repressor and show that minichromosome loci can be used as sites for stable, regulatable transgene expression with low background in non-induced cells. Using these tools, we describe organism-specific constructs for inducible RNA-interference (RNAi) and demonstrate knockdown of multiple essential and non-essential genes. We also show that a minichromosomal site can be exploited to create a stable bloodstream-form line that robustly provides >40,000 independent stable clones per transfection-enabling the production of high-complexity libraries of genome-scale. Finally, we show that modified forms of T. congolense are still infectious, create stable high-bioluminescence lines that can be used in models of AAT, and follow the course of infections in mice by in vivo imaging. These experiments establish a base set of tools to change T. congolense from a technically challenging organism to a routine model for functional genetics and allow us to begin to address some of the fundamental questions about the biology of this important parasite.

Stress Testing Pathology Models with Generated Artifacts

Background: Machine learning models provide significant opportunities for improvement in health care, but their “black-box” nature poses many risks. Methods: We built a custom Python module as part of a framework for generating artifacts that are meant to be tunable and describable to allow for future testing needs. We conducted an analysis of a previously published digital pathology classification model and an internally developed kidney tissue segmentation model, utilizing a variety of generated artifacts including testing their effects. The artifacts simulated were bubbles, tissue folds, uneven illumination, marker lines, uneven sectioning, altered staining, and tissue tears. Results: We found that there is some performance degradation on the tiles with artifacts, particularly with altered stains but also with marker lines, tissue folds, and uneven sectioning. We also found that the response of deep learning models to artifacts could be nonlinear. Conclusions: Generated artifacts can provide a useful tool for testing and building trust in machine learning models by understanding where these models might fail.

Fault Injection in Model-Based System Failure Analysis of Highly Automated Vehicles

The active safety control systems of highly automated vehicles for SAE level 3 and higher are still not fully developed and facing some unresolved issues. The deployment of automated driving systems and the functional safety development present challenges in driver – machine control relationship when there is a system failure or malfunction. The current definition of the product development and controllability classes of the road vehicles functional safety (ISO26262) are not feasible in highly automated vehicles (HAV). This research developed an overview of fault or disturbance injection on the steering system of highly automated model to study the impact of steering system sensors malfunction. The approach was to study the fault propagation using a model-based engineering development in a virtual environment of MATLAB. Subsequently, the steering control system of automated vehicle was developed using an adaptive model predictive control structure to study the control system sensors failures on a system-feature level of the vehicle. It was concluded that the steering wheel angle sensor failure has a significant impact on the planned trajectory of the vehicle and thus it was classified as ASIL D, which represents the highest critical safety component and requires comprehensive safety mechanisms to meet the safety goals of the system. The study also introduced a new criterion for controllability classes suitable for highly automated systems based on the global vehicle position relative of the lane marker lines, to deal with the active safety systems and risk handling strategies. The drivers – vehicle control systems are changing significantly in SAE level 3 automated vehicle and above that driving functions are controlled by the vehicle control systems. This presents human factors challenge in this interactive system with moving to SAE levels 4 and 5. Hence, several human machine interfaces and scenario-based testing are introduced to mitigate any risk or safety uncertainty resulting from control handing-over between the driver and the vehicle control system.

ROS-induced dramatic lipid changes in Arabidopsis

ABSTRACT Objectives: The beneficial role of ROS was probably in promoting intercellular communication by modifying membrane constituents [Liang D. A salutary role of reactive oxygen species in intercellular tunnel-mediated communication. Front Cell Dev Biol. 2018;6:2]. We investigated how the membrane lipids were responding to ROS and ROS inhibitors. Methods: To examine how ROS affected the lipid profiles, we used thin-layer chromatography to characterize lipid profiles in Arabidopsis plants. Then, the confocal microscopy imaging was used to confirm the change of membrane lipid in a plasma membrane marker line exposed to ROS and ROS inhibitors. Results: We found the relative contents of most lipids in H2O2-treated Arabidopsis plants were increased in roots, rather than in shoots. The increased fluorescent signal of membrane marker induced by H2O2 was mainly enriched in the conductive parts of roots. Several ROS inhibitors also strongly affected the lipid profiles. Among them, diethyldithiocarbamate (DDC) can progressively change the lipid profiles with treatment going on. Membrane marker signal was mainly accumulated in the root tips and epidermal cells after treatment by DDC. Discussion: H2O2 may enhance intercellular communication by inducing different lipid species in the conductive parts of roots. The lipid profiles were widely responding to various ROS reagents and might play a role in intercellular signaling.

Targeted Transgene Expression in Rice Using a Callus Strong Promoter for Selectable Marker Gene Control.

Precise expression of a transgene in the desired manner is important for plant genetic engineering and gene function deciphering, but it is a challenge to obtain specific transgene expression free from the interference of the constitutive promoters used to express the selectable marker gene, such as the Cauliflower mosaic virus (CaMV) 35S promoter. So, the solutions to avoid these inappropriate regulations are largely demanded. In this study, we report the characterization of a callus strong promoter (CSP1) in rice and its application for accurate transgene expression. Our results indicate that the high expression of the CSP1 promoter in the callus enables efficient selection of hygromycin equivalent to that provided by the CaMV 35S promoter, whereas its expression in other tissues is low. To evaluate possible leaky effects, the expression of a β-glucuronidase reporter driven by six specific promoters involving hormone signaling, pathogen response, cell fate determination, and proliferation was observed in transgenic rice plants generated by CSP1-mediated selection. Distinct β-glucuronidase expression was found consistently in most of the transgenic lines obtained for each promoter. In addition, we applied these specific marker lines to investigate the root cellular responses to exogenous cytokinin and auxin treatment. The results reveal that the root growth inhibition by cytokinin was differently regulated at high and low concentrations. In summary, we have established the feasibility of using callus-specific promoter-dependent selection to mitigate the transgene misexpression in rice. By enabling efficient transformation, rice plants with reliable transgene expression will be easily acquired for broad applications.

Computer-Aided System Application Value for Assessing Hip Development.

PurposeA computer-aided system was used to semiautomatically measure Tönnis angle, Sharp angle, and center-edge (CE) angle using contours of the hip bones to establish an auxiliary measurement model for developmental screening or diagnosis of hip joint disorders.MethodsWe retrospectively analyzed bilateral hip x-rays for 124 patients (41 men and 83 women aged 20–70 years) who presented at the Affiliated Zhongshan Hospital of Dalian University in 2017 and 2018. All images were imported into a computer-aided detection system. After manually outlining hip bone contours, Tönnis angle, Sharp angle, and CE angle marker lines were automatically extracted, and the angles were measured and recorded. An imaging physician also manually measured all angles and recorded hip development, and Pearson correlation coefficients were used to compare computer-aided system measurements with imaging physician measurements. Accuracy for different angles was calculated, and the area under the receiver operating characteristic (AUROC) curve was used to represent the diagnostic efficiency of the computer-aided system.ResultsFor Tönnis angle, Sharp angle, and CE angle, correlation coefficients were 0.902, 0.887, and 0.902, respectively; the accuracies of the computer-aided detection system were 89.1, 93.1, and 82.3%; and the AUROC curve values were 0.940, 0.956, and 0.948.ConclusionThe measurements of Tönnis angle, Sharp angle, and CE angle using the semiautomatic system were highly correlated with the measurements of the imaging physician and can be used to assess hip joint development with high accuracy and diagnostic efficiency.

Impaired Osteoblastic Differentiation of MSCs Suppresses Normal Hematopoiesis in MDS

Myelodysplastic syndromes (MDS) is a clonal disorder of hematopoietic stem cells (HSCs) characterized by clonal hematopoietic stem cells (HSCs) with cytopenia, morphological abnormalities, genetic alteration, ineffective normal hematopoiesis, and frequent progression to AML. It has long remained unresolved how MDS cells, which are less proliferative, inhibit normal hematopoiesis and eventually come to dominate the bone marrow space. Despite several studies of mesenchymal stem cells (MSCs), one of the principal components of HSC niche supporting normal hematopoiesis, the molecular mechanisms underlying this process remain unclear. In this study, we examined the mechanism by which less-proliferative MDS cells outcompete normal hematopoiesis through the effects on MSCs using serially transplantable Abcg2-induced MDS/AML model we recently generated. The recipient-derived normal BM cells displayed a considerably lower colony output with markedly decreased numbers of the hematopoietic stem progenitor cells (HSPCs) . However, there were no direct effects on the colony-forming ability of the recipient HSPCs co-cultured with MDS/AML cells, indicating that MDS/AML cells inhibited hematopoiesis through alteration of bone marrow microenvironment, such as MSCs, rather than direct interaction between normal and malignant HSCs. We next analyzed histological features of BM specimens. Interestingly, bone sections from the MDS/AML mice showed a reduced trabecular bone and narrowed growth plates. Moreover, micro computed tomography (micro-CT) analysis of the femora showed a significant reduction of the trabecular bone volume in the recipient mice transplanted with the MDS/AML BM cells. We detected decreased bone formation based on the calcein double labeling, but unchanged numbers of the TRAP-positive mononuclear or multinucleated (osteoclastic) cells in the MDS/AML samples, suggesting that the reduced bone volume was caused by suppressed bone formation. The impaired bone formation was also observed in the human MDS patients in terms of lower bone volume and decreased expression of BGLAP, one of osteogenic markers. In line with the above findings, single cell qRT-PCR analyses of mouse MSCs displayed downregulation of a line of osteolineage markers, indicating that MDS/AML cells suppress bone formation through inhibiting osteolineage differentiation of MSCs. Based on the findings, we next examined if re-induction of osteolineage differentiation of the MDS/AML-derived MSCs could rescue the potential of MSCs to support normal hematopoiesis. Importantly, the number of colony-forming cells (CFCs) was significantly restored by inducing differentiation of MDS/AML-derived MSCs toward osteogenic lineage both in vitro and in vivo. These results indicate that the impairment of osteolineage differentiation is the principal cause for an impaired normal hematopoiesis in MDS/AML, and that restoring the supportive niche will be a potential therapeutic option. Since extracellular vesicles (EVs) derived from MDS/AML cells are critical mediators of intercellular communication, we examined the molecular mechanism underlying the dysfunction of MSCs via EVs. As expected, EVs from MDS/AML cells were incorporated into the normal MSCs where osteolineage marker genes were clearly downregulated, and the number of CFCs significantly decreased in the HSPCs co-cultured with MSCs treated by the MDS/AML-derived EVs. Moreover, by comprehensively analyzing microRNAs (miRNAs) enriched in EVs derived from MDS/AML cells, we identified several miRNAs that impaired the differentiation of normal MSCs. These results suggested that miRNAs in EVs derived from MDS/AML cells disrupted the hematopoietic supporting niche through suppressing an osteolineage differentiation of MSCs. Here we uncover a heretofore unrecognized mechanism of bone marrow failure in MDS via the impairment of osteolineage differentiation in MSCs. EVs from MDS cells will be an attractive therapeutic target to restore the supportive niches, such as MSCs, for the remaining normal HSCs. Figure Disclosures No relevant conflicts of interest to declare.

Research on Detecting Moving Objects in Football Match Video

In recent years, detecting moving objects in video has attracted extensive attention in the field of computer vision and pattern recognition. With football becoming one of the most popular sports, analyzing the process of football games is of commercial value and guiding significance. However, the previous research mainly focused on the event detection, and rarely on getting the trajectory of moving objects by directly extracting from the video. This article discussed the method of detecting moving objects in the football match video. We extract the marker lines and build the coordinate system based on LUV color space. And we use the 68 landmarks to recognize the human face and the Kalman filter approach to detect the football. Our research is aimed at helping the audience and football professionals in analyzing the player performance and team strategy.

Cellular Base of Mint Allelopathy: Menthone Affects Plant Microtubules.

Plants can use volatiles for remote suppression of competitors. Mints produce essential oils, which are known to affect the growth of other plants. We used a comparative approach to identify allelopathic compounds from different Mints (genus Mentha, but also including Cat Mint, Nepeta cataria, and Corean Mint, Agastache rugosa, belonging to sisters clades within the Mentheae) using the standard cress germination assay as readout. To understand the mechanism behind this allelopathic effect, we investigated the response of tobacco BY-2 cell lines, expressing GFP-tagged markers for microtubules and actin filaments to these essential oils. Based on the comparison between bioactivity and chemical components, we identified menthone as prime candidate for the allelopathic effect, and confirmed this bioactivity targeted to microtubules experimentally in both, plant cells (tobaccoBY-2), and seedlings (Arabidopsis thaliana). We could show that menthone disrupted microtubules and induced mortality linked with a rapid permeabilization (less than 15 min) of the plasma membrane. This mortality was elevated in a tubulin marker line, where microtubules are mildly stabilized. Our study paves the way for the development of novel bioherbicides that would be environmentally friendly.

Untroubling children’s identity in child protection and welfare assessment through a postconventional analytic

SummaryThe sustaining argument advanced in this paper is that postconventional analysis offers a constructive theoretical lens to contemplate identity in child protection and welfare assessment. Critical application of abridged postconventionalism considers how identity and biodiversity in children and young people, operating across lines of racial, disability, gender and other identity markers, can be understood during informed assessment of needs.FindingsIn the context of transformative postmodern influences, the intention is to provide an aid to students and practitioners who are increasingly impelled to think about traditionally biological-based identity categories, across more abstract lines of social, lingual and cultural construction. Four themes for professional development are presented. First, the complex origins of identity must be appreciated. Second, criticality and openness to the meaning of identity is preferable. Third, progress lies in courteous, sensitive and inclusive practice. Fourth, practitioners should acquire knowledge competency in identity categories.ApplicationsOverall, acceptance of the somewhat fluid and socially contingent nature of identity, without marked rejection of its embodied aspects, can promote more sensitive and inclusive ways of working with children and young people.

On the problem of genetic polymorphism dynamics in Russian cultivars of garden pea (<i>Pisum sativum</i> L.)

Background . Breeding new crops is associated with a risk of decrease (erosion) in genetic polymorphism. It is associated mainly with the fact that only a limited range of pre-existing cultivars and forms are used for breeding new cultivars. The Russian cultivars of garden pea ( Pisum sativum L.) are characterized by a high level of phenotypic variability, while dynamics of polymorphism at DNA level is poorly investigated. Such an investigation is relevant from the point of view of the prospects for further breeding of the crop, and for planning strategies of polymorphism conservation in germplasm collections. Materials and methods . The material used in the study included 18 Russian cultivars bred before 1991, 22 Russian cultivars created later, 40 foreign cultivars, as well as 7 marker lines and wild-growing accessions. These lines were phenotyped and genotyped using DNA markers (14 CAPS, 8 SSR, one mitochondrial and one chloroplast). Variability level was measured as an average Jaccard coefficient resulting from pairwise comparison within each group. Additionally, we analyzed the published characteristics of cultivars included in the State Register in 1994 - 2019. Results . At the phenotypic level, the variability of grain cultivars increased, while a decrease was recorded for vegetable cultivars. A comparison with foreign cultivars has shown that their polymorphism was similar to those of Russian cultivars, except for the vegetable cultivars which are phenotypically less polymorphic than foreign ones. The DNA polymorphism demonstrated a similar tendency, although to a lesser extent. A low level of within-cultivar polymorphism was found. Productivity characteristics of cultivars included into the State Register in the early period (1995 - 2000) and those included later (2016 - 2019) do not differ significantly. Conclusions . There is no clear indication of genetic erosion in Russian cultivars of pea. We also found no reliable increase in the basic agriculturally important characters within the two decades.

Total mesorectal excision with preservation of Denonvilliers' fascia (iTME) based on membrane anatomy

Total mesorectal excision (TME) has been advocated as the golden standard of mid-low rectal cancer surgery for nearly 30 years. However, the complication of postoperative urinary and sexual dysfunctions due to intraoperative nerve injury has yet to be improved. Based on the concept of membrane anatomy, we carried out a systematic study on the important membrane anatomical structure anterior to the rectum--Denonvilliers' fascia. From multiple aspects including anatomy, physiology, histochemistry and surgical practice, we verified the importance of Denonvilliers' fascia for TME surgery in prevention of intraoperative nerve injury and postoperative urogenital dysfunction. Moreover, based on anatomical study of the surgical marker line of Denonvilliers' fascia (Wei's line) and surgical plane, we proved that total mesorectal excision with preservation of Denonvilliers' fascia (iTME) was feasible and practical. Therefore, we conducted a large multicentric randomized controlled trial (RCT). The mid-term result demonstrated that compared with traditional TME surgery, iTME was more effective in reducing the incidence of postoperative urinary and sexual dysfunctions in male patients with mid-low rectal cancer, without sacrifice of short-term tumor radical outcome. We believe that the final RCT result of iTME, based on membrane anatomy, will provide solid evidence for the update of concepts of rectal cancer surgery.