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The widely cultivated medicinal and ornamental plant sage (Salvia officinalis L.) is an evergreen shrub of the Lamiaceae family, native to the Mediterranean. We assembled a high-quality sage genome of 480 Mb on seven chromosomes, and identified a biosynthetic gene cluster (BGC) encoding two pairs of diterpene synthases (diTPSs) that, together with the cytochromes P450 (CYPs) genes located inside and outside the cluster, form two expression cascades responsible for the shoot and root diterpenoids, respectively, thus extending BGC functionality from co-regulation to orchestrating metabolite production in different organs. Phylogenomic analysis indicates that the Salvia clades diverged in the early Miocene. In East Asia, most Salvia species are herbaceous and accumulate diterpenoids in storage roots. Notably, in Chinese sage S. miltiorrhiza, the diterpene BGC has contracted and the shoot cascade has been lost. Our data provide genomic insights of micro-evolution of growth type-associated patterning of specialized metabolite production in plants.

Glioblastoma is the most common primary brain malignancy with limited treatment options. EphA2 is a tumor-associated-antigen overexpressed in glioblastoma. Pre-clinical studies have demonstrated the promise of EphA2-redirected CAR T-cells against glioblastoma. We conduct the first-in-human trial of EphA2-redirected CAR T-cells in patients with EphA2-positive recurrent glioblastoma and report the results of three patients enrolled as the first cohort receiving the starting dosage (1×106 cells/kg). A single infusion of EphA2-redirected CAR T-cells was administrated intravenously, with the lymphodepletion regimen consisting of fludarabine and Cyclophosphamide. In two patients, there was grade 2 cytokine release syndrome accompanied by pulmonary edema, which resolved completely with dexamethasone medication. Except that, there was no other organ toxicity including neurotoxicity. In both the peripheral blood and cerebral-spinal-fluid, we observed the expansion of CAR T-cells which persisted for more than four weeks. In one patient, there was a transit diminishment of the tumor. Among these three patients, one patient reported SD and two patients reported PD, with overall survival ranging from 86 to 181 days. At the tested dose level (1×106 cells/kg), intravenously infusion of EphA2-rediretected CAR T-cells were preliminary tolerable with transient clinical efficacy. Future study with adjusted dose and infusion frequency of CAR T-cells is warranted.Trial Registration NumbersNCT 03423992

In this paper, the low cycle fatigue failure and ratcheting behavior, as well as their interaction of AH32 steel were experimentally investigated under uniaxial cyclic loading. The effects of mean stress, stress amplitude and stress ratio on the low cycle fatigue life and ratcheting strain were discussed. It was found that the ratcheting strain increased while the fatigue life decreased with the increase of mean stress and stress amplitude, and the increasing stress ratio would result in smaller ratcheting and larger fatigue life. Two kinds of failure modes, i.e. low cycle fatigue failure due to crack propagates and ratcheting failure due to large plastic strain will take place respectively. Based on the experimental results, considered the effect of ratcheting on fatigue life, a model with the maximum stress and ratcheting strain rate was proposed. Comparison with the experimental result showed that the new model provided a good prediction for AH32 steel. Keywords: Low cycle fatigue, Ratcheting, Cyclic loading, AH32 steel

Cells of eukaryotic multicellular organisms have inherent heterogeneity. Recent advances in single-cell gene expression studies enable us to explore transcriptional regulation in dynamic development processes and highly heterogeneous cell populations. In this study, using a high-throughput single-cell RNA-sequencing assay, we found that the cells in Arabidopsis root are highly heterogeneous in their transcriptomes. A total of 24 putative cell clusters and the cluster-specific marker genes were identified. The spatial distribution and temporal ordering of the individual cells at different developmental stages illustrate their hierarchical structures and enable the reconstruction of continuous differentiation trajectory of root development. Moreover, we found that each root cell cluster exhibits distinct patterns of ion assimilation and hormonal responses. Collectively, our study reveals a high degree of heterogeneity of root cells and identifies the expression signatures of intermediate states during root cell differentiation at single-cell resolution. We also established a web server (http://wanglab.sippe.ac.cn/rootatlas/) to facilitate the use of the datasets generated in this study.

The present study aims to investigate the risk factors for urosepsis and the diagnostic and prognostic values of the bone morphogenetic protein endothelial cell precursor-derived regulator (BMPER) in patients with urosepsis following ureteroscopic lithotripsy. A total of 305 patients with unilateral ureteral obstruction caused by calculi were included in the study. Patients were divided into three groups, namely, high, medium, and low perfusion pressure groups. The serum C-reactive protein, procalcitonin, lactate (LAC), and BMPER were measured after operation. A logistic regression model was used to assess the risk factors for postoperative urosepsis. The relationships of BMPER with laboratory parameters were explored with a multiple linear regression model. Receiver operating characteristic (ROC) curves were used to diagnosis urosepsis. The cumulative incidence of the adverse events after operation was calculated and compared by log-rank test. Forty-five patients (14.8%) had an episode of urosepsis after operation. Irrigation pressure was an independent risk factor for urosepsis. LAC and sequential organ failure assessment (SOFA) were associated with BMPER after operation. The area under curve value of BMPER for urosepsis was 0.829 (95% confidence interval [CI], 0.773 to 0.884). Uroseptic patients with higher BMPER concentration exhibited more adverse outcome. BMPER possesses valuable discriminative capacity for urosepsis and is a strong predictor of adverse outcome in patients with urosepsis.

Context: Ursolic acid (UA; 3β-hydroxy-urs-12-en-28-oic acid), one of the pentacyclic triterpenoids found in various plants and herbs, possesses some beneficial effects under pathological conditions, including combating hepatic fibrosis. Objective: This study investigates the effects of UA on renal tubulointerstitial fibrosis in vivo and in vitro. Materials and methods: In vivo, 24 male C57BL6 mice were divided into four groups. Eighteen mice were subjected to unilateral ureteral obstruction (UUO) and the remaining six sham-operated mice served as control. UUO mice received either vehicle or UA (50 or 100 mg/kg) by gastric gavage for 6 days. In vitro, HK-2 cells were treated with 10 or 50 μM UA and 10 ng/mL recombinant human transforming growth factor-β1 (TGF-β1). The molecular mechanisms of fibrosis were investigated. Results: UUO induced marked interstitial collagen I and fibronectin deposition and epithelial-mesenchymal transition (EMT), as evidenced by increased α-smooth muscle actin (α-SMA) and decreased E-cadherin. However, UA treatment significantly reduced collagen I and fibronectin accumulation in the fibrotic kidney. UA treatment also decreased α-SMA and preserved E-cadherin in vivo. In vitro, TGF-β1-treated HK-2 cells demonstrated elevated α-SMA, snail1, slug, TGF-β1, and p-smad3, as well as diminished E-cadherin. UA pretreatment prevented E-cadherin loss and diminished α-SMA expression in HK-2 cells. UA downregulated mRNA expression of snail1 and slug. UA also lowered TGF-β1 protein expression and p-Smad3 in HK-2 cells. Conclusions: UA attenuated renal tubulointerstitial fibrosis by inhibiting EMT, and such inhibition may be achieved by decreasing profibrotic factors. UA may be a novel therapeutic agent for renal fibrosis.

The present work aims to investigate the fatigue crack growth behavior of AH-32 steel with the experimental application of CTOD. A series of fatigue crack growth experiments on AH32 steel were conducted by a constant amplitude loading with a single/two tensile overload. The fatigue crack growth behavior of AH32 steel with overload was explored in a detailed manner. The retardation effect of the overload with different overload ratio and overload spacing were quantified by crack growth rate. Moreover, a different relationship between d a / d N and CTOD for all tests was obtained. In a parallel study, the elastic-plastic numerical model was developed to investigate the crack closure effect expressed in the form of crack opening load and CTOD. Experimental results show that the CTOD is able to characterize and correlate the fatigue crack growth at large scale yielding. This work is intended to contribute to a better understanding of the different mechanisms driving fatigue crack growth.

Objective To evaluate long-term functional and survival outcomes of patients with glioma after intraoperative neurophysiologic monitoring (IONM) application. Methods A total of 856 patients with glioma, who underwent tumor resection between October 2010 and March 2016, were included in this retrospective cohort study. All patients were stratified into IONM (439 patients) and non-IONM groups (417 patients). The primary outcome measured was overall survival (OS), and the secondary outcome measured was rate of late neurologic deficits. Analyses were performed using univariate tests and multivariate logistic regression and Cox proportional hazard model. Results The 2 cohorts were well balanced with respect to baseline characteristics. Univariate survival analysis showed longer OS in the IONM group than that in the non-IONM group (P = 0.036), especially in patients with high-grade astrocytic tumor (P = 0.034). The IONM group showed a lower rate of neurologic deficits than did the non-IONM group. Multivariate analysis showed that IONM was a favorable factor of OS (odds ratio, 0.776; P = 0.046) and late neurologic function (odds ratio, 0.583; P = 0.039). Dominant hemispheric and eloquent location of glioma had no association with OS. Conclusions Application of IONM is beneficial to long-term functional and oncologic outcomes of patients with glioma.

Summary Open or accessible regions of the genome are the primary positions of binding sites for transcription factors and chromatin regulators. Transposase-accessible chromatin sequencing (ATAC-seq) can probe chromatin accessibility in the intact nucleus. Here, we describe a protocol to generate ATAC-seq libraries from fresh Arabidopsis thaliana tissues and establish an easy-to-use bioinformatic analysis pipeline. Our method could be applied to other plants and other tissues and allows for the reliable detection of changes in chromatin accessibility throughout plant growth and development. For complete details on the use and execution of this protocol, please refer to Wang et al. (2020)., Graphical Abstract, Highlights • A protocol to generate ATAC-seq libraries from fresh plant tissues • An easy-to-use bioinformatic analysis pipeline for ATAC-seq • Identification of differentially accessible peaks by ATAC-seq, Open or accessible regions of the genome are the primary positions of binding sites for transcription factors and chromatin regulators. Transposase-accessible chromatin sequencing (ATAC-seq) can probe chromatin accessibility in the intact nucleus. Here, we describe a protocol to generate ATAC-seq libraries from fresh Arabidopsis thaliana tissues and establish an easy-to-use bioinformatic analysis pipeline. Our method could be applied to other plants and other tissues and allows for the reliable detection of changes in chromatin accessibility throughout plant growth and development.

This paper proposes a single-stage three-phase quasi-Z-source inverter with strong boost ability and three new kinds of simple boost modified space vector modulation (SVM) strategies. Compared with the traditional quasi-Z-source inverter, the quasi-Z-source inverter with the simple boost modified SVM strategy has the advantages of maximizing the use of zero vector action time, small impedance network volume, low switching loss and voltage stress, and strong boosting ability. The performance parameters of the inverter with different modulation strategies were deeply investigated and compared, including the switching losses of the inverter bridge and the impedance network diode, the loss of the energy storage inductor. The experimental results of the 1.2kVA 96VDC/220V50Hz AC inverter prototype verified the feasibility of the inverter and the correctness of the theoretical analysis, determined a simple boost modified SVM strategy with optimal performance, which lay the key technical foundations on new energy power generation systems such as photovoltaic, wind.

OBJECTIVE: To observe the curative effects of four kinds of plant extracts nasal spray mixture (myrtle oil, hyaluronic acid, fucoidan, anthocyanin) on 161 patients with nasal mucosa dryness, hemorrhae and erosion. METHODS: ① Diagnosis: patients with nasal dryness, erosive and hemorrhae nasal septum mucosa confirmed by patients' complaints and nasal endoscopic test; ② Evaluation: the severity of subjective symptoms was evaluated by VAS score method from 0 to 10 points; the degree of mucosal lesions was divided into 0-8 points by nasal endoscopic test. ③ Intranasal Intranasal administration: 2 times per-day with 2 sprays on each side of the nasal cavity, and no less than 6-8 weeks. In the patients which with allergic rhinitis were also given conventional oral administration of montelukast at the same time. ④ Follow up and curative effect evaluation: VAS and nasal endoscopic test scores that before and after treatment was statistically processed; the patients' own how feels the treatment were evaluated; finally, doctor made the final evaluation based on the combined results of subjective and objective evaluation results before and after treatment, the patients' satisfaction with treatment effect. RESULTS: ① There were significant differences in VAS scores of 161 patients before and after treatment (P < 0.01).② There were statistically significant differences in nasal endoscopic test scores of 104 patients before and after treatment (P < 0.01). ③ Patients' satisfaction survey showed the satisfied with the compliance, tolerance and treatment effect was 95%. CONCLUSION: The mixed nasal spray of four plant extracts can effectively improve the dryness, hemorrhae and erosion of nasal mucosal.

Shoot regeneration is mediated by the sequential action of two phytohormones, auxin and cytokinin. However, the chromatin regulatory landscapes underlying this dynamic response have not yet been studied. In this study, we jointly profiled chromatin accessibility, histone modifications, and transcriptomes to demonstrate that a high auxin/cytokinin ratio environment primes Arabidopsis shoot regeneration by increasing the accessibility of the gene loci associated with pluripotency and shoot fate determination. Cytokinin signaling not only triggers the commitment of the shoot progenitor at later stages but also allows chromatin to maintain shoot identity genes at the priming stage. Our analysis of transcriptional regulatory dynamics further identifies a catalog of regeneration cis-elements dedicated to cell fate transitions and uncovers important roles of BES1, MYC, IDD, and PIF transcription factors in shoot regeneration. Our results, thus, provide a comprehensive resource for studying cell reprogramming in plants and provide potential targets for improving future shoot regeneration efficiency.

Esophageal cancer is still one of the most common cancers in the world. We review the appropriate treatments at different stages of esophageal cancer and also analyze the advantages and disadvantages of these treatments. The prognosis and recovery of different treatment regimens are further discussed. In particular, post-operative complications are the major causes of high mortality derived from the esophageal cancer. Therefore, we particularly discuss the main complications resulting in high mortality after surgery of esophageal cancer, and summarize their risk factors and treatment options. Background As the common cancer, the complications of esophageal cancer after surgery have been not obtained systematic treatment strategy, focusing on treatment regimens based on the different stages of esophageal cancers. Methods and overview This paper systematically summarizes the appropriate treatment strategies for different stages of esophageal cancers, and their advantages and disadvantages. We particularly focus on the postoperative survival rate of patients and postoperative complications, and discuss the causes of high mortality risk factors after surgery. The risk factors of death and corresponding treatment methods are further summarized in this study. Conclusion Postoperative complications is the main cause responsible for the hard cure of esophageal cancers. The existing literatures indicate that postoperative anastomotic fistula is one of the most important complications leading to death, while it has not received much attention yet. We suggest that anastomotic fistula should be detected and dealt with early by summarizing these literatures. It is, therefore, necessary to develop a set of methods to predict or check anastomotic fistula in advance.

MiR-146a has been shown to negatively regulate innate immune, inflammatory response and antiviral pathway, however, its role in the tolerogenic responses remains largely unknown. This study aimed to investigate the role of miR-146a in the OVA-induced allergic inflammation of dendritic cells (DCs).Bone marrow-derived DCs (BMDCs) were treated with OVA (100 µg/ml) for 24 h. MiR-146a expressions were assessed by quantitative RT-PCR. BMDCs were transfected with miR-146a mimics or inhibitor. Cell surface markers were analyzed by flow cytometry. Cytokine levels were determined by ELISA assay. Mixed lymphocyte culture assay was adopted to assess CD4 + T-cell differentiation. The 3' UTR luciferase reporter assay was utilized to determine the miRNA target sequence.OVA treatment significantly up-regulated miR-146a in BMDCs in a dose- and time-dependent manner. In the OVA-treated DCs, overexpression of miR-146a (mimics transfection) down-regulated the surface markers (CD80, CD86) and increased production of anti-inflammatory cytokines TGF-β1 and IL-10 but decreased pro-inflammatory cytokine IL-12. MiR-146a overexpression promoted immature DC to induce regulatory T cells (Treg) differentiation. By contrast, transfection of miR-146a inhibitor into DC exhibited the opposite trends. Notch1 was a direct target of miR-146a, and Notch1 knock-down induced similar effects as miR-146a mimics transfection in BMDCs. Moreover, the effect of miR-146a inhibitor on OVA-induced DC was attenuated by Notch1 knock-down.miRNA-146a promoted tolerogenic properties of DCs, at least partially, through targeting Notch1 signaling.

Backgroud: Lipopolysaccharide (LPS) exposure can induce Leydig cell dysfunction. This study aims to investigate whether adrenomedullin (ADM) rescues the steroidogenic functions of Leydig cells by suppressing TGF-β1 via PI3K/Akt/ GSK-3β/β-catenin signaling. Methods: Rats were given LPS and injected with an adeno-associated virus vector that expressed ADM (Ad-ADM). Plasma concentrations of testosterone and estradiol were measured. Testes were collected and analyzed by gene expression profiling, western blot and immunofluorescence staining. Primary Leydig cells were treated with various concentrations of LPS and ADM. Cell viability and proliferation and medium concentrations of testosterone and estradiol were detected. Gene expression and protein l evels were detected for steroidogenic enzymes, TGF-β1, PI3K, Akt, GSK-3β and β-catenin. Role of ADM in the regulation of TGF-β1 promoter was confirmed by ChIP and Co-IP. Finding: ADM reversed the decreased number of Leydig cells and plasma concentrations of testosterone and estradiol by rescuing the expression of SF-1, 17β-HSD, LRH1, CYP19, StAR, P450scc, 3β-HSD and CYP17. ADM inhibited the LPS-induced cytotoxicity and reversed the cell proliferation by rescuing the expression of Ki-67. ADM also rescued the expression of SF-1, 17β-HSD, LRH1, CYP19, StAR, P450scc, 3β-HSD and CYP17 and the medium concentrations of testosterone and estradiol in LPS-induced Leydig cells. ADM ameliorated LPS-induced TGF-β1 expression in LPS-exposed Leydig cells. ADM enhanced the phosphorylation of PI3K, Akt and GSK-3β but reduced the activation of β-catenin which interacted with TCF4 and then bound to TGF-β1 gene promoter. Interpretation: ADM may rescue the steroidogenic functions of Leydig cells by suppressing TGF-β1 via the PI3K/Akt/GSK-3β/β-catenin signaling. Funding Statement: This study was supported by the National Science Foundation of China, Beijing, China (Grant Nos.: 81402100, 81401190), Natural Science Foundation of Hunan Province, China (Grant Nos.: 2019JJ40269, 2020JJ4542), Foundation for “Young Medical Talents” of Jiangsu Province, China (Grant No.: QNRC2016840), and Key Guidance Project of University of South China, China (Grant No.: USCKF201902K01). Declaration of Interests: The authors declare no competing financial interests with regard to the publication of this paper. Ethics Approval Statement: Procedures involving rats were approved by the Local Ethics Committee for the Care and Use of Laboratory Animals of University of South China and were conducted in accordance with the national guidelines and protocols of the National Institutes of Health.

Additional file 7: Figure S3. Meta-analysis on antral follicle count in non-RCTs: the ultra-long protocol versus long protocol (A), and the ultra-long protocol versus short protocol (B).

Additional file 8: Figure S4. Meta-analysis on basal E2 levels: the ultra-long protocol versus long protocol in RCTs (A) and in non-RCTs (B), and the ultra-long protocol versus short protocol in non-RCTs (C).

Additional file 4: Table S4. Basic characteristics of the included studies.

Additional file 5: Figure S1. Meta-analysis on age: the ultra-long protocol versus long protocol in RCTs (A) and non-RCTs (B).

Background. The role of miR-223-3p in dendritic cells (DCs) is unknown. This study is aimed at investigating the effect of miR-223-3p on the antigen uptake and presentation capacities of DCs and the underlying molecular mechanism. Methods. FITC-OVA antigen uptake and cell surface markers in bone marrow-derived DCs (BMDCs) were analyzed by flow cytometry. BMDCs were transfected with the miR-223-3p mimic or inhibitor. Cytokine levels were determined by ELISA. CD4+ T cell differentiation was determined by mixed lymphocyte culture assay. Results. OVA treatment significantly downregulated miR-223-3p in BMDCs. The miR-223-3p mimic significantly inhibited OVA-induced antigen uptake and surface expression of MHC-II on BMDCs (P<0.01). The miR-223-3p mimic increased TGF-β1 production in OVA-treated DCs (P<0.01). Mixed lymphocyte reaction showed that the miR-223-3p mimic significantly promoted Treg cell differentiation. In addition, the miR-223-3p mimic significantly upregulated CD103 in DCs, indicating the promotion of tolerogenic DCs. The miR-223-3p mimic downregulated Rhob protein in OVA-induced DCs. Rhob knockdown significantly suppressed the ability of FITC-OVA endocytosis (P<0.01) and surface MHC-II molecule expression (P<0.01) in BMDCs, promoting promoted Treg cell differentiation. Mannose receptor (MR) knockdown significantly upregulated miR-223-3p, downregulated Rhob protein in OVA-treated DCs, inhibited the FITC-OVA endocytosis and surface MHC-II expression in BMDCs, and promoted Treg cell differentiation (all P<0.01). Conclusion. These data suggest that miR-223-3p has an inhibitory effect on the antigen uptake and presentation capacities of BMDCs and promotes Treg cell differentiation, which is, at least partially, through targeting MR signaling and Rhob.

Additional file 6: Figure S2. Meta-analysis on BMI in non-RCTs: the ultra-long protocol versus long protocol (A), and the ultra-long protocol versus short protocol (B).