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BackgroundSanda, a martial art that primarily involves punching, kicking, and throwing techniques, requires athletes to maintain high levels of concentration during combat. Sanda principally involves striking the opponent to secure victory, with trauma frequently occurring to the head; however, it remains unclear whether prolonged training enhances or impairs the athletes' attentional capacities. This study aimed to investigate the impact of Sanda training on athletes' attention by employing attention network tests.MethodsA retrospective analysis was conducted on 37 professional Sanda athletes from a certain sports academy; 38 college students from the same institution majoring in physical education were recruited as the control group. Control participants had no prior experience in Sanda training, and the Sanda and control groups were matched in terms of sex, age, and education level. The Attention Network Test (ANT) was administered to both groups to compare differences in efficiency across the alerting, orienting, and executive control networks.ResultsCompared to the control group, the Sanda athletes exhibited significantly higher executive control network efficiency values and executive control network efficiency ratios (P < 0.05). There were no significant differences between the Sanda group and the control group regarding the efficiency values of the alerting and orienting networks (P > 0.05). Additionally, total accuracy and total reaction time between the Sanda athletes and control participants showed no statistically significant differences (P > 0.05).ConclusionSanda practice has detrimental effects on attention, including a decrease in the efficiency of the executive control network and a prolongation of the total reaction time. Therefore, athletes should improve attention training and use head protection to prevent frequent head impacts during training.

Introduction The literature indicates that pandemics significantly impact the mental health of frontline health workers. While the effects of COVID-19 on the mental health of frontline nurses have been studied, their lived experiences remain insufficiently explored. Objective This study aims to investigate the lived experience of nurses who were deployed to support Wuhan during the COVID-19 pandemic. Methods This study adopted a qualitative study design. A purposive sample of fifteen nurses were recruited from a group of nurses who supported Wuhan during COVID-19 pandemic. The data was collected during May and June 2020. Data collection occurred in May and June 2020, employing semistructured interviews conducted via telephone. Interpretative phenomenological analysis (IPA) was utilized to analyze the collected data by two independent researchers. This report follows the COREQ checklist. Results Frontline nurses supporting Wuhan likened their experience to being on a battlefield. Four superordinate themes emerged: (1) mobilization for combating COVID-19; (2) rapid adaption to a dynamic high-stress environment; (3) navigating psychological distress; and (4) the journey home. Conclusion This study offers comprehensive insights into the lived experience of nurses deployed from other provinces to assist COVID patients in Wuhan. The findings indicate that despite facing a variety of challenges, these frontline nurses were capable of rapid adaptation and successfully fulfilled their roles. Recommendations for future preparedness in public health emergencies are provided. Additionally, follow-up research is warranted to explore the long-term effects of frontline experience on the mental health of the nurses and their family members.

Abstract Objective To explore the risk factors for disease progression after initial treatment of type B thymomas using a predictive nomogram model. Methods A single-center retrospective study of patients with type B thymoma was performed. The Cox proportional hazard model was used for univariate and multivariate analyses. Variables with statistical and clinical significance in the multivariate Cox regression were integrated into a nomogram to establish a predictive model for disease progression. Results A total of 353 cases with type B thymoma were retrieved between January 2012 and December 2021. The median follow-up was 58 months (range: 1–128 months). The 10-year progression-free survival (PFS) was 91.8%. The final nomogram model included R0 resection status and Masaoka stage, with a concordance index of 0.880. Non-R0 resection and advanced Masaoka stage were negative prognostic factors for disease progression (p

Animal mitochondrial DNA is usually considered to comply with strict maternal inheritance, and only one mitochondrial DNA haplotype exists in an individual. However, mitochondrial heteroplasmy, the occurrence of more than one mitochondrial haplotype, has recently been reported in some animals, such as mice, mussels, and birds. This study conducted extensive field surveys to obtain representative samples to investigate the existence of paternal inheritance of mitochondrial DNA (mtDNA) in natural fish populations. Evidence of paternal mitochondrial DNA leakage of P. dabryanus was discovered using high-throughput sequencing and bioinformatics methods. Two distinct mitochondrial haplotypes (16,569 bp for haplotype I and 16,646 bp for haplotype II) were observed, differing by 18.83% in nucleotide sequence. Phylogenetic analysis suggests divergence between these haplotypes and potential interspecific hybridization with M. anguillicaudatus, leading to paternal leakage. In natural populations of P. dabryanus along the Yangtze River, both haplotypes are present, with Type I being dominant (75% copy number). Expression analysis shows that Type I has higher expression levels of ND3 and ND6 genes compared to Type II, suggesting Type I’s primary role. This discovery of a species with two mitochondrial types provides a model for studying paternal leakage heterogeneity and insights into mitochondrial genome evolution and inheritance.

【Objective】The light condition is one of the key factors that affect the tissue culture efficiency of Anthurium andraeanum. The effects of different LED light quality ratio and light intensity on different stages of A. andraeanum tissue culture were studied to explore these light conditions suitable for each stage of tissue culture of A. andraeanum and provide an important technical reference for efficient tissue culture of A. andraeanum.【Method】The new A. andraeanum cultivar Fuxing was used as the experimental material, and the effects of different proportions of red and blue LED light ratio and light intensity on callus induction, proliferation culture and strong seedling culture of A. andraeanum Fuxing were analyzed.【Result】The LED red light quality was more suitable for the callus induction of A. andraeanum Fuxing, the 10% R light quality condition with light intensity of 100 lx had the highest callus induction rate of 79.58%, and the callus mass was yellow-green, compact, large in volume and good in state, which was suitable for the callus induction of A. andraeanum Fuxing leaves. The light quality ratio of R ∶ B = 3 ∶ 1 was suitable for callus proliferation culture of A. andraeanum Fuxing, and the proliferation coefficient was the highest (6.44) when the light intensity was 600 lx. Compared with WL light conditions, R ∶ B = 4 ∶ 1 light quality ratio was more suitable for the callus bud differentiation culture of A. andraeanum Fuxing, when the light intensity was 1 000 lx, the number of callus proliferation and differentiation was 19.44, and the buds were short and strong with good growth potential, without yellow leaf phenomenon. The light quality ratio suitable for the tissue culture seedling culture of A. andraeanum Fuxing at the strong seedling stage was R∶B = 4∶1, the light intensity was 900 lx, the number of roots of the strong seedlings of was more, with an average of 5.67, the plant height was 2.88 cm, the stem diameter was 1.44 mm, the leaf length was 11.69 mm, the leaf width was 8.54 mm, the plant leaf was large, the SPAD value was 39.23, the chlorophyll content of the leaf was higher, and the overall growth of the plant was robust without yellow leaf phenomenon.【Conclusion】LED light conditions have an impact on callus induction, proliferation culture, and strong seedling culture of A. andraeanum Fuxing in tissue culture. According to the lighting requirements at different stages of A. andraeanum tissue culture, selecting appropriate LED light quality ratio and light intensity conditions can effectively improve the callus induction rate, proliferation rate and tissue culture seedling production quality of A. andraeanum in tissue culture, and provide technical support for the efficient and standardized production of new A. andraeanum cultivar Fuxing.

IntroductionProviding adolescents with stress management interventions via mobile apps has potential for overcoming barriers to traditional in-person services, such as stigma, cost and travel. However, the effectiveness remains uncertain and engagement level remains low. Therefore, it is essential to understand adolescents’ user experience of such apps, however, such research is scarce. This study aimed to address this research gap by exploring factors affecting Chinese adolescents’ perceived usefulness and engagement of a stress management app, which was developed for them.MethodsA qualitative study design involving focus group interviews and inductive thematic analysis was adopted. A purposive sampling method was employed, resulting in five focus groups (n = 39 adolescents).ResultsTwo themes emerged: (1) mechanism and determinants of usefulness and (2) facilitators and barriers to engagement. The app was found to be helpful in managing chronic and simple stressors by promoting positive behavior, cognition, and physical changes. Relevance to real-life situations, peer support, and planning and monitoring features were found to increase usefulness. Participants suggested adding one-on-one chat support for managing acute stressors. Multimedia, logical content arrangement, combining psychoeducation and skills training, gamification, customization, and an appealing user interface were engaging factors for adolescents, whilst text-heavy content, pedagogical and monotonous tones, technical issues were found to disengage adolescents.ConclusionStress management apps should involve simple and evidence-based coping skills training, target adolescents’ real-life problems, promote positive peer influence, address both chronic and acute stressors. Additionally, such apps should have logical arrangement of content, be interactive and customizable, and involve multimedia and gamification features to engage adolescents.

Abstract Background Stress in adolescence is associated with adverse mental health outcomes. Coping resources have been proved by literature to have buffering effects on the impact of stress on mental health. It is imperative to understand the stress and coping strategies of adolescents. However, to date, there has been a scarce of qualitative examination of stress and coping strategies in adolescents in a Chinese population. Objectives This study aimed to understand the stress experience and coping strategies of high school students in China. Methods This study adopted a qualitative design involving three focus group interviews. A purposive sampling method was used to recruit high school students who were enrolled in grades 10 to 11, and their teachers, at a Chinese high school which resulted in 20 students and 9 teacher participants. Data were analysed using inductive thematic analysis. Results A total of 4 themes were identified: (i) sources of stress; (ii) impacts of stress (iii) coping strategies used by students; and (iv) recommendations for stress management programs. Students experienced excessive stress in their daily lives. The primary source of stress came from high expectations for academic achievement. Other sources of stress were peer relationships and family issues. The stress had negative impacts on students’ emotions, sleep, study, and mental wellbeing. The students demonstrated various coping strategies, with the most common being avoidant coping. Students and teachers agreed that the coping strategies were not effective in reducing stress in the long run and that more coping skills training was needed. Conclusions This study is the first to assess the perceptions of Chinese high school students and their teachers regarding adolescent stress experiences and coping strategies. Chinese high school students experienced significant stress in their daily lives and demonstrated unhelpful coping strategies. Participants demonstrated consensus that they did not have the skills to cope. There is a demonstrated need for interventions that focus on increasing coping skills in this population.

The electro-hydrostatic actuator (EHA) is a new type of high-performance servo actuator that originated in the field of aerospace, and it is gradually becoming a common basic component for various types of large equipment. A miniature EHA, mainly composed of a micro two-dimensional (2D) piston pump and a brushless DC motor, is designed in this article by simplifying the system structure. This paper analyzes the structure and working principle of this EHA and establishes the mathematical models of the brushless DC motor, micro two-dimensional pump, and hydraulic cylinder. Field-oriented control (FOC) is used to drive the brushless DC motor, and the models of the controller are established in Simulink. Furthermore, the models of the mechanical and hydraulic systems of the miniature EHA are established in AMESim. In addition to this, a prototype of this miniature EHA was fabricated in this paper and an experimental platform was built for experiments. In the joint simulation environment, the rise time of the EHA system at 6000 r/min is 0.158 s and the frequency response amplitude attenuation to −3 dB has a bandwidth of 20 Hz. On the other hand, the constructed miniature EHA prototype was dynamically characterized to obtain a rise time of 0.242 s at 6000 r/min and a bandwidth of 13 Hz. In this paper, the feasibility of the design scheme of the miniature EHA system is verified, and its excellent dynamic properties are verified with simulation and experiment.

Modern anthuriums, Anthurium andraeanum (Hort.) are among the most popular flowering plants and widely used for interior decoration. Their popularity is largely attributed to the exotic spathes with different colors. Previous studies have reported color development in red spathe cultivars, but limited information is available on key genes regulating white and green colored spathes. This study analyzed anthocyanin, chlorophyll, and carotenoid contents as well as transcript differences in spathes of eight cultivars that differed in spathe colors ranging from red to white and green. Results showed that increased expression of a transcription factor AaMYB2 was associated with elevated levels of anthocyanin in spathes, but decreased expression of AaMYB2 and increased expression of AaLAR (leucoanthocyanidin reductase) and AaANR (anthocyanidin reductase) were accompanied with the accumulation of colorless proanthocyanidin, thus the white spathe. As to the green colored spathe, chlorophyll content in the green spathe cultivar was substantially higher than the other cultivars. Correspondingly, transcripts of chlorophyll biosynthesis-related genes AaHemB (porphobilinogen synthase) and AaPor (protochlorophyllide oxidoreductase) were highly upregulated but almost undetectable in white and red spathes. The increased expression of AaHemB and AaPor was correlated with the expression of transcription factor AaMYB124. Subsequently, qRT-PCR analysis confirmed their expression levels in nine additional cultivars with red, white, and green spathes. A working model for the formation of white and green spathes was proposed. White colored spathes are likely due to the decreased expression of AaMYB2 which results in increased expression of AaLAR and AaANR, and the green spathes are attributed to AaMYB124 enhanced expression of AaHemB and AaPor. Further research is warranted to test this working model.

Purpose – Eating disorders (EDs) is a major health condition affecting 9% of the global population and 10% of those with EDs lost their lives as a result. Text-based telehealth interventions (TTIs) seem to provide a low-cost and convenient treatment option; however, the evidence is scarce. This study aimed to synthesize evidence relating to the use of TTIs for the management of EDs. Design/methodology/approach – Five databases were searched published between January 2020 and May 2019. The authors used keywords relating to telehealth and EDs. The authors used Joanna Briggs Institute's (JBI's) critical appraisal instrument to assess the methodology quality of included studies. Findings – Fifteen studies were included in this mix-method systematic review and assessed for methodology quality. Email, web-based texting, text-messaging and online chat room were used as mode for deliver healthcare for patients with EDs. In the treatment phase, all studies (nine studies; n = 860 participants) showed effectiveness (for RCTs) and usefulness (for non-RCT studies). In the aftercare phase (six studies; n = 364 participants), the results regarding the effectiveness of TTIs were mixed. Two studies showed effectiveness whilst four studies did not find statistically significant change of ED outcomes. Research limitations/implications – The qualities of these studies varied; firstly, 66% (n = 10) of the studies were non-randomized studies (e.g. single-arm trial, case report) with small samples. Moreover, one-fourth (n = 4) of the studies did not use validated instruments or indicate the instrument. Also, half (n = 7) of the studies used TTIs as adjunct to face-to-face treatment or bigger online treatment, it is hard to make conclusion that the changes were due to TTIs' effect. In addition, follow-up rate is not satisfactory, thus results should be interpreted cautiously. Practical implications – TTIs seem to be promising for management of EDs, particularly in the treatment phase. This provides an important treatment option for health practitioners and people with EDs as an alternative or in adjunct with face-to-face services. Originality/value – This is the first review to synthesis the use of TTIs for ED management.

【Objective】The study aims to screen out the types, concentrations and application methods of plant growth regulators that can effectively promote early flowering and improve flowering uniformity of Gymnocalycium baldianum, and provide technical support for the regulation of flowering period of G. baldianum.【Method】With uniform seed balls of G. baldianum as test materials, the effects of different concentrations (100, 200, 400 and 800 mg/L) of paclobutrazol, uniconazole, indoleacetic acid and gibberellin on its flowering was explored.【Result】Spraying 100-800 mg/L paclobutrazol, uniconazole and indoleacetic acid solutions had no significant difference in the colour of seed balls, seed ball diameter increment, number of flowers and flowering rate. However, these three plant growth regulators could promote the early flowering of G. baldianum within a appropriate concentration range. The treatment of 100 mg/L paclobutrazol had the best result, which bloomed about 9.0 days earlier than that under water treatment. It could reach the full blooming period within 11.7 days, which was about 20.0 days shorter than that under water treatment, and the flowering had better uniformity. While the 100-800 mg/L gibberellin solution had an inhibitory effect on the flowering of G. baldianum, however, it could promote vegetative growth. The ball size, the number of thorns in the thorn seat, the thorn length, and the mass of the thorn seat all increased, and the colour of the seed ball was greener.【Conclusion】The study results can provide technological support for promoting the early flowering of G. baldianum and improving the uniformity of flowering. At the same time, it can improve the ornamental characters of G. baldianum according to individual needs, and improve its ornamental value, which has great significance to increase market share furtherly.

【Objective】Tissue culture is one of the important ways for rapid propagation of Aglaonema commutatum, the factors affecting leaf color in tissue culture were studied and explored to maintain the redness of leaf color, and to provide technical references for further stabilizing efficient tissue culture technology and regulating leaf color of A. commutatum with red leaves.【Method】Taking A. commutatum 'Guanghuahong' with red leaves as experimental material, the effects of proliferation medium, proliferation mode, light intensity and light quality on leaf color of A. commutatum were studied.【Result】The improved MS medium supplemented with 0.5 g/L K2SO4 was used for proliferation culture, and the proliferation and differentiation seedlings were more robust, with brighter leaf color, and the transplanting seedling rate was 90.00%.The leaf color of tissue culture seedlings cultured by the proliferation of clustered buds was more stable than that cultured by the proliferation of callus, and the transplanting seedling rate was higher. In the stages of proliferation and rooting, when the light intensity was 4 000 lx, the red plaque area increased, and the transplanting seedling rate was higher (88.89%). Under the light quality condition of red light : blue light = 3∶7, the lowest total content of chlorophyll in leaves was 0.037 mg/g, and the highest relative content of anthocyanin was 0.399 Units/g, and the leaves were red and brilliant.【Conclusion】Using the improved proliferation medium, adopting the proliferation mode of clustered buds, adjusting the light intensity and changing the red and blue light quality ratio can effectively maintain the redness of leaves and improve the production efficiency of tissue culture.

ObjectivesThis study aimed to determine the efficacies of the Coping Camp app in reducing stress, depression, and anxiety and improving stress-coping behaviours and mental health wellbeing. Additionally, feasibility and acceptability of Coping Camp were evaluated.MethodsIn this unblinded cluster RCT, 540 participants from two high schools in China were randomly assigned to the Coping Camp intervention (n = 6 classes; 275 students) or treatment as usual (n = 5 classes; 265 students) at the class level. Coping Camp was an automated self-help app, consisting of 11 sessions delivered over 11 weeks, with primary outcomes including perceived stress, depression, anxiety, stress-coping behaviours, and mental health well-being. All outcomes were assessed at baseline, post-intervention (11 weeks), and follow-up (19 weeks), with efficacy analysed using linear mixed models and feasibility/acceptability measured by a 5-point Likert scale and qualitative feedback.ResultsAt post-intervention and follow-up assessments, 75.4% and 81.7% of participants respectively attended. On average, participants logged in for 8.56 out of 11 sessions. Compared to the control group, the intervention group had significant reductions in levels of perceived stress (p = 0.01, d = 0.15 at T1; p < 0.001, d = 0.18 at T2), anxiety (p = 0.11; d = 0.08 at T1; p = 0.01; d = 0.13 at T2) and depression (p = 0.04, d = 0.11 at T1; p = 0.05, d = 0.10 at T2) but did not have a greater increase in stress-coping behaviours (p = 0.10 at T1; p = 0.97 at T2) or mental health wellbeing (p = 0.93 at T1; p = 0.08 at T2). The average ratings for each session were above 4, and qualitative feedback showed that most participants found the intervention to be "great," "good," and "useful."ConclusionsThe Coping Camp is feasible, acceptable and effective in stress management among Chinese school adolescents.

Dual-energy X-ray absorptiometry and the Fracture Risk Assessment Tool are recommended tools for osteoporotic fracture risk evaluation, but are underutilized. Here, the authors present an opportunistic tool to identify fractures, predict bone mineral density and evaluate fracture risk using plain pelvis and lumbar spine radiographs.

Abstract Background Peroxisome proliferator-activated receptor alpha (PPARα) is associated with diabetic retinopathy (DR), and the underlying mechanism is still unclear. Aim of this work was to investigate the mechanism of PPARα in DR. Methods Human retinal capillary pericytes (HRCPs) were treated with high glucose (HG) to induce DR cell model. DR mouse model was established by streptozotocin injection, and then received 5-Aza-2-deoxycytidine (DAC; DNA methyltransferase inhibitor) treatment. Hematoxylin–eosin staining was performed to assess retinal tissue damage. PPARα methylation was examined by Methylation-Specific PCR. Flow cytometry and DCFH-DA fluorescent probe was used to estimate apoptosis and reactive oxygen species (ROS). The interaction between DNA methyltransferase-1 (DNMT1) and PPARα promoter was examined by Chromatin Immunoprecipitation. Quantitative real-time PCR and western blot were performed to assess gene and protein expression. Results HG treatment enhanced the methylation levels of PPARα, and repressed PPARα expression in HRCPs. The levels of apoptotic cells and ROS were significantly increased in HRCPs in the presence of HG. Moreover, DNMT1 was highly expressed in HG-treated HRCPs, and DNMT1 interacted with PPARα promoter. PPARα overexpression suppressed apoptosis and ROS levels of HRCPs, which was rescued by DNMT1 up-regulation. In DR mice, DAC treatment inhibited PPARα methylation and reduced damage of retinal tissues. Conclusion DNMT1-mediated PPARα methylation promotes apoptosis and ROS levels of HRCPs and aggravates damage of retinal tissues in DR mice. Thus, this study may highlight novel insights into DR pathogenesis.

Abstract Background Pulmonary large-cell neuroendocrine carcinoma (pLCNEC) is a very rare malignancy originating from the lung and bronchus, and its biological behaviour, clinical diagnosis, treatment and prognosis are poorly understood. Thus, the clinical characteristics and surgical treatment-related prognostic factors of this rare disorder must be explored. Results The clinical data of 59 patients (48 males and 11 females) who were treated by surgery and diagnosed with pLCNEC by postoperative pathology at Peking Union Medical College Hospital from April 2004 to April 2019 were analysed retrospectively. The median patient age was 62 years (38–79 years), and the median duration of disease was 2 months (0.5–18 months). Compared with other lung malignancies, pLCNEC lacks specific clinical symptoms and imaging features, and preoperative biopsy pathology is often insufficient to confirm the diagnosis. The corresponding numbers of patients who were classified into stages I, II, III and IV according to the postoperative pathological tumour-nodal-metastasis stage were 25, 12, 15 and 7, respectively. The median overall survival was 36 months (0.9–61.1 months). The 1-year, 3-year and 5-year survival rates were 76.3%, 49% and 44.7%, respectively. The tumour stage exerted a significant effect on survival (Cox multivariate analysis p

Abstract Background Heat resistance is a common characteristic of harpins, a class of proteins found in Gram-negative bacteria, which may be related to the stability of coiled-coil (CC) structure. The CC structure is a ubiquitous protein folding and assembly motif made of α-helices wrapping around each other forming a supercoil. Specifically, whether the stability of the CC structure near to N-terminus of four selected harpin proteins from Xanthomonas (hereafter referred to as Hpa1) would influence their characteristics of heat resistance was investigated. We used bioinformatics approach to predict the structure of Hpa1, used the performance of hypersensitive response (HR)-induction activity of Hpa1 and circular dichroism (CD) spectral analyses to detect the relationship between the stability of the CC structure of Hpa1 and heat resistance. Results Each of four-selected Hpa1 has two α-helical regions with one in their N-terminus that could form CC structure, and the other in their C-terminus that could not. And the important amino acid residues involved in the CC motifs are located on helices present on the surface of these proteins, indicating they may engage in the formation of oligo mericaggregates, which may be responsible for HR elicitation by harpins and their high thermal stability. Increased or decreased the probability of forming a CC could either induce a stronger HR response or eliminate the ability to induce HR in tobacco after high temperature treatment. In addition, although the four Hpa1 mutants had little effect on the induction of HR by Hpa1, its thermal stability was significantly decreased. The α-helical content increased with increasing temperature, and the secondary structures of Hpa1 became almost entirely α-helices when the temperature reached 200 °C. Moreover, the stability of the CC structure near to N-terminus was found to be positively correlated with the heat resistance of Hpa1. Conclusions The stability of the CC structure might sever as an inner drive for mediating the heat resistance of harpin proteins. Our results offer a new insight into the interpretation of the mechanism involved in the heat resistance of harpin protein and provide a theoretical basis for further harpin function investigations and structure modifications.

Triplophysa tenuis is an important indigenous fish in the Xinjiang Tarim River. In this study, we collected 120 T. tenuis individuals from 8 T. tenuis populations in the Tarim River. Through genotyping-by-sequencing (GBS), a total of 582,678,756 clean reads were generated for all the genotypes, and after quality filtering, 595,379 SNPs were obtained for the population genetic analyses. Multiple genetic parameters showed that the 8 T. tenuis populations had high genetic diversity. Phylogenetic tree analysis indicated that all T. tenuis individuals were divided into five branches, the individuals from the north of Tarim River were grouped into cluster 1 (SF and WS) and cluster 3 (DWQ, TKX, and KZE), while the AETS, WLWT and LF individuals from the south of Tarim River were clustered into cluster 2. The result was consistent with the admixture analysis, which supported that the 8 T. tenuis populations were clustered into three subgroups. Furthermore, the pairwise FST values and genetic distance indicated that there was a large genetic differentiation between WS and other T. tenuis populations. Collectively, this study provides valuable genome-wide data for the conservation of natural T. tenuis populations in the Tarim River.

Background The management of ground‐glass opacities (GGOs) depends mainly on personal experience. In clinical practice, benign GGOs are not rare in resected specimens, for which operations may be avoided. We retrospectively compared the clinical features of resected GGOs to identify differential diagnostic characteristics. Methods Among 1456 patients with suspected malignant GGOs who underwent surgical resection, 105 patients (35 with benign GGOs and 70 matched controls with malignant GGOs) were included. Clinical characteristics, including demographics and radiologic, surgical and pathologic characteristics, were collected. Results The smoking index (P = 0.044), frequency of coughing (P = 0.026), GGO size (P = 0.003), size change during follow‐up (P = 0.011), location (P = 0.022), presence of air bronchogram sign (P = 0.004), distance to the pleura (P = 0.021) and positron emission tomography/computed tomography (PET/CT) appearance (P = 0.003) showed significant differences between the benign and malignant groups. Pathologically, the resected benign GGOs included focal fibrosis (17), inflammation or infection (seven), lymphoproliferative disorder (one), hamartoma (three), inflammatory myofibroblastic tumor (two), hemangioma or vascular malformation (two), endometriosis (two) and pulmonary cyst (one). Conclusions A higher smoking index, coughing, larger size, similar or increased size during follow‐up, location in the upper and middle lobes, air bronchogram sign on CT, lesion margin to pleura distance over 1 cm, and malignant tendency on PET/CT reports were associated with malignant GGOs. Relatively active surgical interventions could be considered for GGOs highly suspected of malignancy.

Abstract Background Harpins are proteins secreted by the type III secretion system of Gram-negative bacteria during pathogen–plant interactions that can act as elicitors, stimulating defense and plant growth in many types of non-host plants. Harpin-treated plants have higher resistance, quality and yields and, therefore, harpin proteins may potentially have many valuable agricultural applications. Harpins are characterized by high thermal stability at 100 °C. However, it is unknown whether harpins are still active at temperatures above 100 °C or whether different temperatures affect the activity of the harpin protein in different ways. The mechanism responsible for the heat stability of harpins is also unknown. Results We identified a novel harpin, HpaXpm, from the cassava blight bacteria Xanthomonas phaseoli pv. manihotis HNHK. The predicted secondary structure and 3-D structure indicated that the HpaXpm protein has two β-strand domains and two major α-helical domains located at the N- and C-terminal regions, respectively. A phylogenetic tree generated using the maximum likelihood method grouped HpaXpm in clade I of the Hpa1 group along with harpins produced by other Xanthomonas spp. (i.e., HpaG-Xag, HpaG-Xcm, Hpa1-Xac, and Hpa1Xm). Phenotypic assays showed that HpaXpm induced the hypersensitive response (HR), defense responses, and growth promotion in non-host plants more effectively than Hp1Xoo (X. oryzae pv. oryzae). Quantitative real-time PCR analysis indicated that HpaXpm proteins subjected to heat treatments at 100 °C, 150 °C, or 200 °C were still able to stimulate the expression of function-related genes (i.e., the HR marker genes Hin1 and Hsr203J, the defense-related gene NPR1, and the plant growth enhancement-related gene NtEXP6); however, the ability of heat-treated HpaXpm to induce HR was different at different temperatures. Conclusions These findings add a new member to the harpin family. HpaXpm is heat-stable up to 200 °C and is able to stimulate powerful beneficial biological functions that could potentially be more valuable for agricultural applications than those stimulated by Hpa1Xoo. We hypothesize that the extreme heat resistance of HpaXpm is because the structure of harpin is very stable and, therefore, the HpaXpm structure is less affected by temperature.

The diploids and tetraploids of cyprinid loach Misgurnus anguillicaudatus coexist in many natural habits. The tetraploids generally exhibit superior growth performance and delayed gonadal development compared with diploids. To investigate the regulation mechanism of growth superiority and ovarian development in tetraploids, we first conducted a long-term culture experiment and histological observation. The tetraploids exhibited a higher growth performance and delayed ovarian development. Meanwhile, at a genetic level, an average of 6891 differentially expressed genes (DEGs) between diploids and tetraploids were identified from the brain, pituitary, liver, gonad and muscle using the RNA-seq method. Among these DEGs, as expected, some candidate genes, including IGF family genes, somatostatin, leptin, cyp19a1b, gthα, lhβ and fshβ, were detected, which play critical roles in the regulation of growth and gonad development of fish. In particular, the genes related to GH/IGF axis and growth factors, signal transduction, gonadal hormone and appetite were significantly increased in tetraploids. The clustering analyses of the key candidate genes showed that most key genes were up-regulated in the pituitary and gonad of tetraploids instead of other tissues. The dynamics of these key genes provide valuable genetic evidence for clarifying the growth superiority and delayed gonadal development of tetraploids. Moreover, this study also provides some clues for unveiling the genetic superiority of polyploidy species in other phenotypes.

Background: Atezolizumab, a high-affinity engineered human anti–PD-L1 antibody, has produced a clinical benefit for patients with advanced non–small-cell lung cancer (NSCLC). However, associated with T-cell regulation, the immunomodulatory effect of PD-L1 blockade and its biomarker in peripheral immunity remains elusive.Methods: In a prospective cohort with 12 Chinese advanced NSCLC patients who received atezolizumab 1,200 mg every 3 weeks as a second-line treatment, blood samples were obtained before and 6 weeks after atezolizumab initiation, and when disease progression was confirmed. Patients were classified into a response or progression group according to response evaluation criteria in solid tumors (RECIST) 1.1. Fresh peripheral blood mononuclear cells (PBMCs) from patients were stained with antihuman CD3, CD8, and PD-1 antibodies for flow cytometry analysis. T-cell receptor (TCR)-β chains of CD8+ T cells were analyzed by next-generation sequencing (NGS) at the deep level. Diversity, clonality, and similarity of TCR have been calculated before and after treatment in both groups.Results: Clonal expansion with high PD-1 expression was detected in all patients’ peripheral CD8+ T cells before the treatment of atezolizumab. Unlike the progression group, the diversity of TCR repertoire and singletons in the TCRβ pool increased over time with atezolizumab administration, and the TCR repertoire dynamically changes in the response group. The percentage of CD8+ PD-1high terminal exhausted T cells declined in the response group after the PD-L1 blockade. Two patterns of TCR changes among patients who received PD-L1–targeted immunotherapy were observed.Conclusions: Deep sequencing of the T-cell receptors confirmed the existence of CD8+ PD-1high T cells with an exhaustion phenotype in Chinese NSCLC patients. Our study demonstrated that efficient anti–PD-L1 therapy could reshape the TCR repertoire for antitumor patients. Furthermore, singleton frequency may help us select patients who are sensitive to anti–PD-L1 immunotherapy.

Background: Crizotinib has potent anti-tumor activity in patients with advanced MET-amplified non-small cell lung cancer (NSCLC). However, the therapeutic effect is still not satisfying. Thus, developing approaches that improve the efficacy of crizotinib remains a significant challenge. Methods: MET-amplified NSCLC cell lines were treated with crizotinib and cyclosporine A (CsA). Cell viability was determined by MTS assay. The changes of apoptosis, cell cycle and calcineurin-Erk pathways were assessed by western blot. Xenograft mouse model, primary human NSCLC cells and hollow fiber assays were utilized to confirm the effects of CsA. Findings: We demonstrated that CsA significantly increased the anti-tumor effect of crizotinib on multiple MET-amplified NSCLC cells in vitro and in vivo. Mechanistically, crizotinib treatment led to the activation of Ca2+-calcineurin (CaN)-Kinase suppressor of Ras 2 (KSR2) signaling, resulting in Erk1/2 activation and enhanced survival of cancer cells. CsA effectively blocked CaN-KSR2-Erk1/2 signaling, promoting crizotinib-induced apoptosis and G2/M arrest. Similarly, pharmacologic or genetic inhibition of Erk1/2 also enhanced crizotinib-induced growth inhibition in vitro. Xenograft studies further confirmed that CsA or Erk1/2 inhibitor PD98059 enhanced the anti-cancer activity of crizotinib through inhibition of CaN-Erk1/2 axis. The results were also validated by primary human NSCLC cells in vitro and hollow fiber assays in vivo. Interpretation: This study provides preclinical evidences that combination therapy of CsA and crizotinib is a promising approach for targeted treatment of MET-amplified lung cancer patients. Fund: This work was supported by the National Natural Science Foundation of China, the Key Projects of Natural Foundation of Zhejiang Province, the Ten thousand plan youth talent support program of Zhejiang Province, the Zhejiang Natural Sciences Foundation Grant, and the Zhejiang medical innovative discipline construction project-2016. Keywords: Cyclosporine A, Erk1/2, Crizotinib, Lung cancer, Calcium

Abstract. Whether regional lymph node involvement exerts significant effect on the prognosis still remains obscure for pancreatic neuroendocrine tumors. To clarify this association and identify predictors for lymph node involvement, we studied the data of patients aged >18 years with regional lymph node involvement histologically confirmed pancreatic neuroendocrine tumors from 2004 to 2014 in the Surveillance, Epidemiology, and End Results database (http://seer.cancer.gov/about). We evaluated Lymph node involvement as a prognostic factor by Cox regression. We reduced 9 variables of demographic and tumor characteristics to 5 potential predictors using least absolute shrinkage and selection operator (LASSO) regression model. We further constructed a lymph node involvement model by logistic regression. The model was verified by the verification set, and the visual expression of the model was realized by a nomogram. A total of 1545 cases of pancreatic neuroendocrine tumors were included in our study. Lymph node positivity was significantly associated with disease-specific survival (P

With the rapid growth of geo-tagged social media data, it has become feasible to explore topics across different areas through text mining and geographical visualization. However, the visual elements of social media data always overlap with each other in the map view, which largely disturbs visual perception of semantic features and their geographical distribution. Thus, it is of great significance to reduce the visual clutter of large-scale social media data, and enhance the visibility of semantic features across local areas. In this paper, we utilize a doc2vec model to transform geo-tagged social media data into high-dimensional vectors, and the semantic correlation can be easily characterized in the dimensionality reduction space. Aiming at the reduction of visual clutter of geographical visualization, a dual-objective blue noise sampling model is proposed to select a subset of social media data, by means of which both the semantic correlation and spatial distribution of large scale social media data are well retained. A rich set of visual designs are implemented enabling users to evaluate the sampled results from multiple perspectives and explore the changes of semantic features across areas, such as heatmap, word cloud and text stream. The effectiveness and validity of the proposed visualization system are further demonstrated through case studies and expert reviews.

A sensitive simultaneous electroanalysis of phytohormones indole-3-acetic acid (IAA) and salicylic acid (SA) based on a novel copper nanoparticles-chitosan film-carbon nanoparticles-multiwalled carbon nanotubes (CuNPs-CSF-CNPs-MWCNTs) composite was reported. CNPs were prepared by hydrothermal reaction of chitosan. Then the CuNPs-CSF-CNPs-MWCNTs composite was facilely prepared by one-step co-electrodeposition of CuNPs and CNPs fixed chitosan residues on modified electrode. Scanning electron microscope (SEM), transmission electron microscopy (TEM), selected area electron diffraction (SAED), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and linear sweep voltammetry (LSV) were used to characterize the properties of the composite. Under optimal conditions, the composite modified electrode had a good linear relationship with IAA in the range of 0.01–50 μM, and a good linear relationship with SA in the range of 4–30 μM. The detection limits were 0.0086 μM and 0.7 μM (S/N = 3), respectively. In addition, the sensor could also be used for the simultaneous detection of IAA and SA in real leaf samples with satisfactory recovery.

Neuroinflammation contributes to delayed (secondary) neurodegeneration following traumatic brain injury (TBI). Tumor necrosis factor receptor-associated factor 6 (TRAF6) signaling may promote post-TBI neuroinflammation, thereby exacerbating secondary injury. This study investigated the pathogenic functions of TRAF6 signaling following TBI in vivo and in vitro. A rat TBI model was established by air pressure contusion while lipopolysaccharide (LPS) exposure was used to induce inflammatory-like responses in cultured astrocytes. Model rats were examined for cell-specific expression of TRAF6, NF-κB, phosphorylated (p)-NF-κB, MAPKs (ERK, JNK, and p38), p-MAPKs, chemokines (CCL2 and CXCL1), and chemokine receptors (CCR2 and CXCR2) by immunofluorescence, RT-qPCR, western blotting, and ELISA, for apoptosis by TUNEL staining, and spatial cognition by Morris water maze testing. These measurements were compared between TBI model rats receiving intracerebral injections of TRAF6-targeted RNAi vector (AAV9-TRAF6-RNAi), empty vector, MAPK/NF-κB inhibitors, or vehicle. Primary astrocytes were stimulated with LPS following TRAF6 siRNA or control transfection, and NF-κB, MAPKs, chemokine, and chemokine receptor expression levels evaluated by western blotting and ELISA. TRAF6 was expressed mainly in astrocytes and neurons of injured cortex, peaking 3 days post-TBI. Knockdown by AAV9-TRAF6-RNAi improved spatial learning and memory, decreased TUNEL-positive cell number in injured cortex, and downregulated expression levels of p-NF-κB, p-ERK, p-JNK, p-p38, CCL2, CCR2, CXCL1, and CXCR2 post-TBI. Inhibitors of NF-κB, ERK, JNK, and p38 significantly suppressed CCL2, CCR2, CXCL1, and CXCR2 expression following TBI. Furthermore, TRAF6-siRNA inhibited LPS-induced NF-κB, ERK, JNK, p38, CCL2, and CXCL1 upregulation in cultured astrocytes. Targeting TRAF6-MAPKs/NF-κB-chemokine signaling pathways may provide a novel therapeutic approach for reducing post-TBI neuroinflammation and concomitant secondary injury.

To reveal the global picture and emerging evolution trends with sufficiently large literature data of RC structure’s service life prediction, especially the time to corrosion initiation, the scientometric analysis on the corresponding evolution trends was performed by using visualization software CiteSpace and VOSviewer in this paper. First, the application of CiteSpace and VOSviewer and retrieval strategy for data collection were described. And then, information visualization analysis was carried out based on the papers related to RC structure’s service life evaluation from publications number evolution, journal distribution, and authors’ contribution. Finally, document cocitation network, cooccurring keywords network, and timeline view of keywords network were conducted to discuss the research hotspots on time to corrosion initiation evaluation of reinforcement. Research results reveal that the number of publications on RC structure’s service life evaluation reached its peak in 2011. Besides, Structural Safety is the journal that makes the most significant contributions, and Professor Li CQ is the author with the most contribution score. Moreover, the knowledge body of the time to corrosion initiation prediction consists of six clusters; the high-citation articles in this field mainly focus on the multiple-parameters model and probabilistic reliability method.

The distribution of Triplophysa yarkandensis is restricted to Xinjiang’s Tarim River basin. We collected 119 T. yarkandensis samples from nine geographic populations in the Tarim River basin and utilized the RAD-seq method for SNP genotyping. In this study, a total of 164.81 Gb bases were generated with the Illumina platform, and 129,873 candidate SNPs were obtained with the Stacks pipeline for population genetic analyses. High levels of genetic diversity were detected among nine populations. The AMOVA results showed that the majority of genetic variations originated from among populations (FST = 0.67), and the pairwise FST values ranged from 0.4579 to 0.8736, indicating high levels of genetic differentiation among these populations. The discriminate analysis of principal components (DAPCs) and neighbor joining (NJ) tree revealed that the nine populations could be separated into two clusters (i.e., south and north populations), and modest genetic differentiation between south and north populations was observed, while the individuals from several populations were not clustered together by geographical location. The evidence of two genetic boundaries between south and north populations (except TTM) was supported by barrier analysis. The Bayesian skyline plotting indicated that T. yarkandensis populations in the Tarim River basin had not experienced genetic bottlenecks, and the effective population size remained stable. This study first clarified the genetic diversity and differentiation of T. yarkandensis populations in the Tarim River basin, and it provided valuable molecular data for conservation and management of natural populations.

Editor's Note: this Article has been retracted; the Retraction Note is available at https://doi.org/10.1038/s41598-021-82870-5 .

ABSTRACT The topmouth gudgeon, Pseudorasbora parva (Temminck & Schlegel, 1842), is a small cyprinid fish that inhabits the littoral zones of freshwater habitats throughout Asia and Europe. It is regarded as an invasive species in Europe, but in its native range, in China, as food and as an environmental bio-indicator. In this study, the embryonic and larval development of P. parva was investigated for the first time. The eggs of P. parva are transparent, adhesive and elliptical. The mean size of fertilized eggs was (1.63×1.31) ± 0.04 mm. From fertilization to hatching, embryonic development could be divided into eight stages and 34 phases, and the incubation period lasted for 109.25 hours at 24 ± 1 °C. Newly hatched larvae were 4.1 ± 3 mm in length, and the yolk absorption was completed within six days after hatching. The first and second swim bladders formed at the third and ninth day, respectively. The pectoral fin formed before the hatching stage, followed by the caudal, dorsal, anal and ventral fin formation after hatching. About 20 days after hatching, the morphology of the fry was similar to the adult fish. These findings provide a basis for determining the complete ontogeny of P. parva, as well as facilitate the management and utilization of this fish.

Potassium (K+) is one of the essential macronutrients required for plant growth and development, and the maintenance of cellular K+ homeostasis is important for plants to adapt to abiotic stresses and growth. However, the mechanism involved has not been understood clearly. In this study, we demonstrated that AtUNC-93 plays a crucial role in this process under the control of abscisic acid (ABA). AtUNC-93 was localized to the plasma membrane and mainly expressed in the vascular tissues in Arabidopsis thaliana. The atunc-93 mutants showed typical K+-deficient symptoms under low-K+ conditions. The K+ contents of atunc-93 mutants were significantly reduced in shoots but not in roots under either low-K+ or normal conditions compared with wild type plants, whereas the AtUNC-93-overexpressing lines still maintained relatively higher K+ contents in shoots under low-K+ conditions, suggesting that AtUNC-93 positively regulates K+ translocation from roots to shoots. The atunc-93 plants exhibited dwarf phenotypes due to reduced cell expansion, while AtUNC-93-overexpressing plants had larger bodies because of increased cell expansion. After abiotic stress and ABA treatments, the atunc-93 mutants was more sensitive to salt, drought, osmotic, heat stress and ABA than wild type plants, while the AtUNC-93-overexpressing lines showed enhanced tolerance to these stresses and insensitive phenotype to ABA. Furthermore, alterations in the AtUNC-93 expression changed expression of many ABA-responsive and stress-related genes. Our findings reveal that AtUNC-93 functions as a positive regulator of abiotic stress tolerance and plant growth by maintaining K+ homeostasis through ABA signaling pathway in Arabidopsis.

The co-existence of several ploidy types in natural populations makes the cyprinid loach Misgurnus anguillicaudatus an exciting model system to study the genetic and phenotypic consequences of ploidy variations. A first step in such effort is to identify the specific ploidy of an individual. Currently popular methods of karyotyping via cytological preparation or flow cytometry require a large amount of tissue (such as blood) samples, which can be damaging or fatal to the fishes. Here, we developed novel microsatellite markers (SSR markers) from M. anguillicaudatus and show that they can effectively discriminate ploidy using samples collected in a minimally invasive way. Specifically, we generated whole genome transcriptomes from multiple M. anguillicaudatus using the Illumina paired-end sequencing. Approximately 150 million raw reads were assembled into 76,544 non-redundant unigenes. A total of 8,194 potential SSR markers were identified. We selected 98 pairs with more than five tandem repeats for further assays. Out of 45 putative EST-SSR markers that successfully amplified and harbored polymorphism in diploids, 11 markers displayed high variability in tetraploids. We further demonstrate that a set of five EST-SSR markers selected from these are sufficient to distinguish ploidy levels, by first validating them on 69 reference specimens with known ploidy levels and then subsequently using fresh-collected 96 ploidy-unknown specimens. The results from EST-SSR markers are highly concordant with those from independent flow cytometry analysis. The novel EST-SSR markers developed here should facilitate genetic studies of polyploidy in the emerging model system M. anguillicaudatus.

HpaXm produced by the cotton leaf blight bacterium Xanthomonas citri subsp. malvacearum is a novel harpin elicitor of the induced hypersensitive response (HR) in tobacco. We investigated whether fragments of HpaXm, compared with fragments of Hpa1Xoo, are sufficient for HR or plant growth promotion (PGP) elicitation using four synthetic peptides (HpaXm35-51, HpaXm10-39, Hpa1Xoo36-52 and Hpa1Xoo10-40). We also heated the fragments to determine the heat tolerance of the functional fragments. HpaXm35-51 and Hpa1Xoo36-52 induced hypersensitive response (HR). Bursts of reactive oxygen intermediates (ROI) induced by HpaXm35-51 and Hpa1Xoo36-52 were earlier and stronger than those induced by HpaXm and Hpa1Xoo. In plants treated with HpaXm35-51 or Hpa1Xoo36-52, the expression of the HR marker genes Hin1 and Hsr203J and the active oxygen metabolism related gene AOX were significantly upregulated. These findings suggest that the predicted α-helical structures of the HpaXm35-51 and Hpa1Xoo36-52 fragments are crucial for HR. PGP result by soaking seeds in unheated/heated HpaXm10-39 or Hpa1Xoo10-40 solution prior to transfer, which obviously enhances root growth and the aerial parts of plants. The PGP related gene NtEXP6 was greatly enhanced when plants were sprayed with a solution of HpaXm10-39 or Hpa1Xoo10-40; heated fragment treatments induced higher levels of NtEXP6 expression than unheated HpaXm fragments. In addition, HR marker genes induced by the heated fragments had lower expression levels than when induced with unheated HpaXm fragments. Moreover, the expression levels of HR marker genes and PGP related genes induced by treatment with Hpa1Xoo fragments before or after heating were the opposite of those induced by HpaXm fragments. Different functional fragments of harpin and different harpins with the same functional region have different degrees of heat tolerance. Therefore, the heat resistance of harpin is conservative, but the degree of heat tolerance of the functional fragments is specific.

Identifying the dispersal pathways of an invasive species is useful for adopting the appropriate strategies to prevent and control its spread. However, these processes are exceedingly complex. So, it is necessary to apply new technology and collect representative samples for analysis. This study used Approximate Bayesian Computation (ABC) in combination with traditional genetic tools to examine extensive sample data and historical records to infer the invasion history of the red swamp crayfish, Procambarus clarkii, in China. The sequences of the mitochondrial control region and the proPOx intron in the nuclear genome of samples from 37 sites (35 in China and one each in Japan and the USA) were analyzed. The results of combined scenarios testing and historical records revealed a much more complex invasion history in China than previously believed. P. clarkii was most likely originally introduced into China from Japan from an unsampled source, and the species then expanded its range primarily into the middle and lower reaches and, to a lesser extent, into the upper reaches of the Changjiang River in China. No transfer was observed from the upper reaches to the middle and lower reaches of the Changjiang River. Human-mediated jump dispersal was an important dispersal pathway for P. clarkii. The results provide a better understanding of the evolutionary scenarios involved in the rapid invasion of P. clarkii in China.

Non–small cell lung cancer is one of the most common types of cancer, and the prognosis of non–small cell lung cancer is still poor. Recent evidence has proved that long noncoding RNA is involved in tumorigenesis. For non–small cell lung cancer, the expression profile of long noncoding RNA has been studied. Here, we identified a novel long noncoding RNA TRPM2-AS from published dataset and found TRPM2-AS is widely upregulated in non–small cell lung cancer tissues compared with adjacent non-tumor tissues. Higher expression level of TRPM2-AS was correlated with higher TNM stages and larger tumor size. Patients with high TRPM2-AS expression level had poor survival than those with low TRPM2-AS level. We silenced TRPM2-AS by small interfering RNA and found that cell proliferation was significantly inhibited after knockdown of TRPM2-AS. Annexin V/propidium iodide staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay confirmed that cell apoptosis increased after TRPM2-AS knockdown. Further experiments showed that silence of TRPM2-AS upregulated SHC1 and silence of SHC1 partially reversed cell apoptosis after TRPM2-AS knockdown. In summary, the novel long noncoding RNA TRPM2-AS upregulated in non–small cell lung cancer, and downregulation of TRPM2-AS promotes apoptosis in vitro.

Epidermal growth factor (EGF) is a potent regulator of cell function in many cell types. In mammals, the EGF/EGFR system played an important role in both pituitary physiology and pathology. However, it is not clear about the pituitary action of EGF in lower vertebrates. In this study, using grass carp as a model, we found that EGF could stimulate NK3R mRNA and protein expression through pituitary ErbB1 and ErbB2 coupled to MEK/ERK and PI3K/Akt/mTOR pathways. In addition, EGF could also induce pituitary somatolactin α (SLα) secretion and mRNA expression in a dose- and time-dependent manner in vivo and in vitro. The stimulatory actions of EGF on SLα mRNA expression were also mediated by PI3K/Akt/mTOR and MEK/ERK pathways coupled to ErbB1 and ErbB2 activation. Our previous study has reported that neurokinin B (NKB) could also induce SLα secretion and mRNA expression in carp pituitary cells. In the present study, interestingly, we found that EGF could significantly enhance NKB-induced SLα mRNA expression. Further studies found that NK3R antagonist SB222200 could block EGF-induced SLα mRNA expression, indicating an NK3R requirement. Furthermore, cAMP/PKA inhibitors and PLC/PKC inhibitors could both abolish EGF- and EGF+NKB-induced SLα mRNA expression, which further supported that EGF-induced SLα mRNA expression is NK3R dependent.

Cuticular wax covers aerial organs of plants and functions as the outermost barrier against non-stomatal water loss. We reported here the functional characterization of the Glossy1(GL1)-homologous gene OsGL1-3 in rice using overexpression and RNAi transgenic rice plants. OsGL1-3 gene was ubiquitously expressed at different level in rice plants except root and its expression was up-regulated under ABA and PEG treatments. The transient expression of OsGL1-3-GFP fusion protein indicated that OsGL1-3 is mainly localized in the plasma membrane. Compared to the wild type, overexpression rice plants exhibited stunted growth, more wax crystallization on leaf surface, and significantly increased total cuticular wax load due to the prominent changes of C30-C32 aldehydes and C30 primary alcohols. While the RNAi knockdown mutant of OsGL1-3 exhibited no significant difference in plant height, but less wax crystallization and decreased total cuticular wax accumulation on leaf surface. All these evidences, together with the effects of OsGL1-3 on the expression of some wax synthesis related genes, suggest that OsGL1-3 is involved in cuticular wax biosynthesis. Overexpression of OsGL1-3 decreased chlorophyll leaching and water loss rate whereas increased tolerance to water deficit at both seedling and late-tillering stages, suggesting an important role of OsGL1-3 in drought tolerance.

Misgurnus anguillicaudatus has several natural ploidy types. To investigate whether nuclear polyploidy have an impact on mitochondrial DNA (mtDNA), the complete mitochondrial genomes (mitogenomes) of five distinct ploidy M. anguillicaudatus (natural diploid, triploid, tetraploid, pentaploid and hexaploid), which were collected in central China, were sequenced and analyzed. The five mitogenomes share the same gene arrangement and have similar gene size, base composition and codon usage pattern. The most variable regions of the mitogenome were the protein-coding genes, especially the ND4L (5.39% mutation rate). Most variations occurred in tetraploids. The phylogenetic tree showed that the tetraploid M. anguillicaudatus separated early from other ploidy loaches. Meanwhile, the mitogenomes from pentaploids, and hexaploids have the closest phylogenetic relations, but far from that of tetraploids, implying that pentaploids and hexaploids could not be formed from tetraploids, possibly from the diploids and triploids. The genus Misgurnus species were divided into two divergent inter-genus clades, and the five ploidy M. anguillicaudatus were monophyletic, which support the hypotheses about the mitochondrial introgression in loach species.

Colorectal cancer (CRC) is still the third most common cancer and the second most common causes of cancer-related death around the world. Metformin, a biguanide, which is widely used for treating diabetes mellitus, has recently been shown to have a suppressive effect on CRC risk and mortality, but not all laboratory studies suggest that metformin has antineoplastic activity. Here, we investigated the effect of metformin and AMPK activator AICAR on CRC cells proliferation. As a result, metformin did not inhibit cell proliferation or induce apoptosis for CRC cell lines in vitro and in vivo. Different from metformin, AICAR emerged antitumor activity and sensitized anticancer effect of 5-FU on CRC cells in vitro and in vivo. In further analysis, we show that AMPK activation may be a key molecular mechanism for the additive effect of AICAR. Taken together, our results suggest that metformin has not antineoplastic activity for CRC cells as a single agent but AMPK activator AICAR can induce apoptosis and enhance the cytotoxic effect of 5-FU through AMPK activation.

BackgroundTraditional Chinese medicine (TCM) has been widely applied for cancer care in China. There have been a large number of controlled clinical studies published in Chinese literature, yet no systematic searching and analysis has been done. This study summarizes the current evidence of controlled clinical studies of TCM for cancer.MethodsWe searched all the controlled clinical studies of TCM therapies for all kinds of cancers published in Chinese in four main Chinese electronic databases from their inception to November 2011. We bibliometrically analyzed the included studies and assessed the reporting quality.ResultsA total of 2964 reports (involving 253,434 cancer patients) including 2385 randomized controlled trials and 579 non-randomized controlled studies were included. The top seven cancer types treated were lung cancer, liver cancer, stomach cancer, breast cancer, esophagus cancer, colorectal cancer and nasopharyngeal cancer by both study numbers and case numbers. The majority of studies (72%) applied TCM therapy combined with conventional treatment, whilst fewer (28%) applied only TCM therapy in the experimental groups. Herbal medicine was the most frequently applied TCM therapy (2677 studies, 90.32%). The most frequently reported outcome was clinical symptom improvement (1667 studies, 56.24%) followed by biomarker indices (1270 studies, 42.85%), quality of life (1129 studies, 38.09%), chemo/radiotherapy induced side effects (1094 studies, 36.91%), tumor size (869 studies, 29.32%) and safety (547 studies, 18.45%). Completeness and adequacy of reporting appeared to improve with time.ConclusionsData from controlled clinical studies of TCM therapies in cancer treatment is substantial, and different therapies are applied either as monotherapy or in combination with conventional medicine. Reporting of controlled clinical studies should be improved based on the CONSORT and TREND Statements in future. Further studies should address the most frequently used TCM therapy for common cancers and outcome measures should address survival, relapse/metastasis and quality of life.

Rice at reproductive stage is more sensitive to environmental changes, and little is known about the mechanism of heat response in rice panicle. Here, using rice microarray, we provided a time course gene expression profile of rice panicle at anther developmental stage 8 after 40°C treatment for 0 min, 20 min, 60 min, 2 h, 4 h, and 8 h. The identified differentially expressed genes were mainly involved in transcriptional regulation, transport, cellular homeostasis, and stress response. The predominant transcription factor gene families responsive to heat stress were Hsf, NAC, AP2/ERF, WRKY, MYB, and C(2)H(2). KMC analysis discovered the time-dependent gene expression pattern under heat stress. The motif co-occurrence analysis on the promoters of genes from an early up-regulated cluster showed the important roles of GCC box, HSE, ABRE, and CE3 in response to heat stress. The regulation model central to ROS combined with transcriptome and ROS quantification data in rice panicle indicated the great importance to maintain ROS balance and the existence of wide cross-talk in heat response. The present study increased our understanding of the heat response in rice panicle and provided good candidate genes for crop improvement.