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Abstract Background Fanconi anemia (FA) is the most common inherited bone marrow failure (BMF) syndrome with 22 related genes identified. The ALDH2 rs671variant has been proved related to accelerate the progression of BMF in FA patients. The phenotype and genetic basis of Chinese FA patients have not been investigated yet. Methods We analyzed the 22 FA-related genes of 63 BMF patients suspected to be FA. Clinical manifestations, morphological and cytogenetic feathers, ALDH2 genotypes, treatment, and outcomes of the definite cases were retrospectively studied. Results A total of 21 patients were confirmed the diagnosis of FA with the median age of BMF onset was 4-year-old. The number of patients manifested as congenital malformations and growth retardation were 20/21 and 14/21, respectively. BM dysplasia and cytogenetic abnormalities were found in 13/20 and 8/19 patients. All the patients with abnormal karyotypes also manifested as BM dysplasia or had evident blasts. Thirty-five different mutations were identified involving six genes and including twenty novel mutations. FANCA mutations contributed to 66.67% of cases. Eight patients harboring ALDH2-G/A genotype have a significantly younger age of BMF onset (p = 0.025). Within the 19 patients adhering to continuous follow-up, 15 patients underwent hematopoietic stem cell transplantations (HSCTs). During the 29 months of follow-up, 8/19 patients died, seven of which were HSCT-related, and one patient who did not receive HSCT died from severe infection. Conclusions The phenotypic and genetic spectrum of Chinese FA patients is broad. Bone marrow dysplasia and cytogenetic abnormalities are prevalent and highly consistent. The overall outcome of HSCTs is disappointing. Nationwide multicenter studies are needed for the rarity and adverse outcome of this disease.

The role of neurons and T cells in glioma progression remains poorly understood. Here the authors show that midkine-dependent activation of a neuron-T cell-microglia axis promotes the growth of optic pathway gliomas.

Selection for new single-flowered amaryllis with cultivated wild species or hybrids has continued for more than 200 years. However, our basic knowledge of the extent of genetic variation within the single-flowered amaryllis is still not sufficient, which is very important for the ongoing improvement of the double-flower type in China. In this study, a total of 23 screened sequence-related amplified polymorphism (SRAP) markers were used to assess the genetic diversity and population structure of 82 cultivated accessions. It produced 244 clearly amplified fragments, of which 242 (99.2%) were polymorphic. A higher level of genetic diversity was observed, with estimates of Nei’s diversity index (H) and the Shannon information index (I) of 0.344 and 0.513, respectively. Genetic distance- and structure-based analyses mainly clustered all accessions into two or five subgroups, indicating the complexity of the hybrid pedigree of single-flowered amaryllis. To effectively explore and preserve the novel variation in these 82 genetic resources, we further established an initial core collection with 15 accessions with the goal of maximizing the SRAP alleles. This preliminary core set retained 100% of the SRAP variation with greater genetic diversity and heterogeneity (H = 0.355, and I = 0.529). Our research provided insight into the genetic structure of modern hybrids of single-flowered amaryllis, from which an easily manageable amaryllis core collection was identified, facilitating future breeding progress for double-flowered amaryllis.

Regulatory T (Treg) cells are critical in preventing aberrant immune responses. Posttranscriptional control of gene expression by microRNA (miRNA) has recently emerged as an essential genetic element for Treg cell function. Here, we report that mice with Treg cell-specific ablation of miR-142 (hereafter Foxp3CremiR-142fl/fl mice) developed a fatal systemic autoimmune disorder due to a breakdown in peripheral T-cell tolerance. Foxp3CremiR-142fl/fl mice displayed a significant decrease in the abundance and suppressive capacity of Treg cells. Expression profiling of miR-142-deficient Treg cells revealed an up-regulation of multiple genes in the interferon gamma (IFNγ) signaling network. We identified several of these IFNγ-associated genes as direct miR-142-3p targets and observed excessive IFNγ production and signaling in miR-142-deficient Treg cells. Ifng ablation rescued the Treg cell homeostatic defect and alleviated development of autoimmunity in Foxp3CremiR-142fl/fl mice. Thus, our findings implicate miR-142 as an indispensable regulator of Treg cell homeostasis that exerts its function by attenuating IFNγ responses.

The neuroscience community has developed many convolutional neural networks (CNNs) for the early detection of Alzheimer’s disease (AD). Population graphs are thought of as non-linear structures that capture the relationships between individual subjects represented as nodes, which allows for the simultaneous integration of imaging and non-imaging information as well as individual subjects’ features. Graph convolutional networks (GCNs) generalize convolution operations to accommodate non-Euclidean data and aid in the mining of topological information from the population graph for a disease classification task. However, few studies have examined how GCNs’ input properties affect AD-staging performance. Therefore, we conducted three experiments in this work. Experiment 1 examined how the inclusion of demographic information in the edge-assigning function affects the classification of AD versus cognitive normal (CN). Experiment 2 was designed to examine the effects of adding various neuropsychological tests to the edge-assigning function on the mild cognitive impairment (MCI) classification. Experiment 3 studied the impact of the edge assignment function. The best result was obtained in Experiment 2 on multi-class classification (AD, MCI, and CN). We applied a novel framework for the diagnosis of AD that integrated CNNs and GCNs into a unified network, taking advantage of the excellent feature extraction capabilities of CNNs and population-graph processing capabilities of GCNs. To learn high-level anatomical features, DenseNet was used; a set of population graphs was represented with nodes defined by imaging features and edge weights determined by different combinations of imaging or/and non-imaging information, and the generated graphs were then fed to the GCNs for classification. Both binary classification and multi-class classification showed improved performance, with an accuracy of 91.6% for AD versus CN, 91.2% for AD versus MCI, 96.8% for MCI versus CN, and 89.4% for multi-class classification. The population graph’s imaging features and edge-assigning functions can both significantly affect classification accuracy.

A reliable assessment and design of engineering structures requires appropriate estimation and consideration of different sources of uncertainty. The randomness of seismic ground motion is one major uncertainty that needs to be considered from the perspective of performance-based earthquake engineering. To properly account for this uncertainty and its corresponding effect on pile foundations, a stochastic seismic response analytical framework based on a probability density evolution method and a stochastic ground motion model is proposed for the nonlinear stochastic seismic response analysis of pile foundations. A three-dimensional finite element model of a reinforced concrete pile with a large diameter embedded in the soil foundation is firstly established and calibrated using the full-scale lateral load test results from the literature, in which the nonlinear behavior of soil, the soil–pile interaction, the concrete damaged plasticity, and the steel yielding of the pile are properly modeled. Then, the calibrated three-dimensional numerical model is employed as an illustrative example for the stochastic seismic response analysis using the proposed framework. The mean, standard deviation, and probability density function of pile settlement and the dynamic reliabilities of the pile concerning various performance requirements are obtained in the present study. The settlements of the pile show great variability under the excitation of various stochastic ground motions, and the maximum mean value of the pile settlements is about 8 mm. The changing shape of the settlement probability density functions in every moment demonstrates that it is unreasonable to use assumed probabilistic distribution models to characterize the seismic responses of pile foundations during a seismic reliability analysis. Based on the proposed method, it is found that the dynamic reliabilities of the selected reinforced concrete pile concerning four different performance levels are 0, 0.1293, 0.8247, and 1, respectively. The proposed stochastic seismic response analytical method in the present study can provide a proper and comprehensive way to quantify various uncertainties and their corresponding effects on the seismic performance of pile foundations. It can also be used to estimate the actual reliability level of pile foundations that are designed by the current codes when they are subjected to seismic loads.

Parkinson’s disease (PD) is one of the most common neurodegenerative disorders. However, its cellular and molecular mechanisms still wrap in the mist. This is partially caused by the absence of appropriate animal models mimicking sporadic PD that constitutes the majority of cases. Previously, we reported that a cysteine protease, asparagine endopeptidase (AEP), is activated in an age-dependent manner, and cleaves α-synuclein in the brain of sporadic PD patients. The AEP-derived α-synuclein 1-103 fragment is required for the pathogenesis of PD. Thus, we designed and characterized a novel transgenic mouse line expressing α-synuclein 1-103 (designated N103 mice). This model shows an abundant accumulation of pathological α-synuclein in the central nervous system, loss of dopaminergic neurons in the substantia nigra, and progressive striatal synaptic degeneration. The N103 mice also manifest age-dependent PD-like behavioral impairments. Notably, the mice show weight loss and constipation, which are the common non-motor symptoms in PD. The RNA-sequencing analysis found that the transcriptomics pattern was extensively altered in N103 mice. In conclusion, the N103 mouse line, as a brand-new tool, might provide new insights into PD research.

The road transport sector in megacities is confronted with pressing local air pollution and carbon dioxide (CO2) control issues. To determine effective policy instruments for saving energy and the co-control of air pollutants and CO2, several mainstream measures were examined and compared in Chongqing's road transport sector from 2017 to 2035. An integration assessment framework was developed by combining the Long-range Energy Alternatives Planning (LEAP) system and a set of quantitative methods for evaluating the co-benefits of emission reductions (including the air pollutant equivalent (APeq), co-control coordinate system, and pollutant reduction cross-elasticity (Elsa/b)). Results showed that the shifting transportation modes scenario presented the most significant potential for energy-saving and emission reductions, reducing energy use by 30.9% and air pollutants and CO2 emissions by approximately 27–32% compared with the business as usual (BAU) scenario in 2035. The improving energy efficiency scenario also provided significant co-benefits for reducing air pollutants and CO2 emissions. Nevertheless, the promoting alternative fuel scenario may increase fine particulate matter (PM2.5) emissions by 2.2% compared to BAU in 2035 under the cleanness of regional electricity in 2017. Our findings suggest that the shifting transportation modes were effective measures to reduce air pollutants and CO2 in the short term synergistically, and highlighted the importance of cleaner electricity generation to develop electric vehicles in the medium and long term.

BackgroundRheumatoid arthritis (RA) is a chronic systemic autoimmune disorder characterized by inflammatory cell infiltration, leading to persistent synovitis and joint destruction. The pathogenesis of RA remains unclear. This study aims to explore the immune molecular mechanism of RA through bioinformatics analysis.MethodsFive microarray datasets and a high throughput sequencing dataset were downloaded. CIBERSORT algorithm was performed to evaluate immune cell infiltration in synovial tissues between RA and healthy control (HC). Wilcoxon test and Least Absolute Shrinkage and Selection Operator (LASSO) regression were conducted to identify the significantly different infiltrates of immune cells. Differentially expressed genes (DEGs) were screened by “Batch correction” and “RobustRankAggreg” methods. Functional correlation of DEGs were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Candidate biomarkers were identified by cytoHubba of Cytoscape, and their diagnostic effectiveness was predicted by Receiver Operator Characteristic Curve (ROC) analysis. The association of the identified biomarkers with infiltrating immune cells was explored using Spearman’s rank correlation analysis in R software.ResultsTen significantly different types of immune cells between RA and HC were identified. A total of 202 DEGs were obtained by intersection of DEGs screened by two methods. The function of DEGs were significantly associated with immune cells. Five hub genes (CXCR4, CCL5, CD8A, CD247, and GZMA) were screened by R package “UpSet”. CCL5+CXCR4 and GZMA+CD8A were verified to have the capability to diagnose RA and early RA with the most excellent specificity and sensitivity, respectively. The correlation between immune cells and biomarkers showed that CCL5 was positively correlated with M1 macrophages, CXCR4 was positively correlated with memory activated CD4+ T cells and follicular helper T (Tfh) cells, and GZMA was positively correlated with Tfh cells.ConclusionsCCL5, CXCR4, GZMA, and CD8A can be used as diagnostic biomarker for RA. GZMA-Tfh cells, CCL5-M1 macrophages, and CXCR4- memory activated CD4+ T cells/Tfh cells may participate in the occurrence and development of RA, especially GZMA-Tfh cells for the early pathogenesis of RA.

Parkinson's disease (PD) is one of the most common neurodegenerative diseases, which is characterized by the loss of dopaminergic neurons in the nigrostriatal pathway. Synaptic dysfunction impairs dopamine turnover and contributes to the degeneration of dopaminergic neurons. However, the molecular mechanisms underlying synaptic dysfunction and dopaminergic neuronal vulnerability in PD are not clear. Here, we report that synaptojanin 1 (SYNJ1), a polyphosphoinositide phosphatase concentrated at nerve terminals, is a substrate of a cysteine proteinase, asparagine endopeptidase (AEP). SYNJ1 is cleaved by the cysteine proteinase AEP at N599 in the brains of PD patients. AEP-mediated cleavage of SYNJ1 disrupts neuronal phosphoinositide homeostasis and causes synaptic dysfunction. Overexpression of the AEP-generated fragments of SYNJ1 triggers synaptic dysfunction and the degeneration of dopaminergic neurons, inducing motor defects in the α-synuclein transgenic mice. Blockage of AEP-mediated cleavage of SYJN1 alleviates the pathological and behavioral defects in a mouse model of PD. Our results demonstrate that the fragmentation of SYNJ1 by AEP mediates synaptic dysfunction and dopaminergic neuronal degeneration in PD.

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases. Pathologically, it is characterized by the aberrant aggregation of α-synuclein (α-syn) in neurons. Clinical evidence shows that patients with hypercholesterolemia are more likely to get PD, while lovastatin users have a lower risk of suffering from it. In this study, we investigated the effects of lovastatin on the aggregation and phosphorylation of α-syn in vitro. Our results demonstrate that α-syn preformed fibrils induce the phosphorylation and aggregation of α-syn in HEK293 cells stably transfected with α-syn-GFP and SH-SY5Y cells as well, which could be attenuated by in a concentration-dependent manner. Besides, lovastatin inhibited oxidative stress, histone acetylation, and the activation of casein kinase 2 (CK2). Collectively, lovastatin alleviates α-syn aggregation and phosphorylation in cellular models of synucleinopathy, indicating its potential value of being adopted in the management of PD.

The genetic diversity and population structure of 38 commercial accessions of double-flowered amaryllis (Hippeastrum hybridum) from the Netherlands and South Africa were evaluated using sequence-related amplified polymorphism (SRAP) markers. Thirty SRAP primer pairs produced 294 loci, of which 263 (89.16%) were polymorphic. A relatively high level of genetic diversity was observed, with estimates of Nei’s diversity index (H) and the Shannon information index (I) of 0.2719 and 0.4158, respectively. Additional genetic distance- and STRUCTURE-based analyses clustered all accessions into two or four subgroups based mostly on origin or color. The genetic differentiation between/among countries and inferred groups was significant, with Fst values ranging from 0.083-0.194%. Accessions from the Netherlands showed higher genetic variation than those from South Africa. Several accessions, such as Aphrodite, are recommended for future programs employing selective hybridization with the goal of expanding the color range. The results of the present study provide appropriate information applicable to designing effective breeding programs for double-flowered amaryllis.

Polarized human airway epithelia at an air-liquid interface (HAE-ALI) are an in vitro

In this paper, a high sensitivity fiber temperature sensor based on surface plasmon resonance is designed and studied. In the simulation, the single mode fiber is polished to remove most of the cladding, and then gold and silver films are added. Finally, it is embedded in the heat shrinkable tube filled with a thermo-optic coefficient liquid for curing. The numerical simulation results show that the sensing characteristics are sensitive to the remaining cladding thickness of the fiber, the thickness of the gold film and the thickness of the silver film. When the thermo-optic coefficient of the filling liquid is −2.8 × 10−4/°C, the thickness of the gold film, the thickness of the silver film and the thickness of the remaining cladding of the fiber are 30 nm, 20 nm and 1 μm, respectively. The sensitivity of the sensor designed in this paper can reach −6 nm/°C; this result is slightly higher than that of similar research in recent years. It will have a promising application prospect in flexible wearable temperature sensors, smart cities and other fields.

Patients often undergo consolidation allogeneic hematopoietic stem cell transplantation (allo-HSCT) to maintain long-term remission following chimeric antigen receptor (CAR) T-cell therapy. Comparisons of safety and efficacy of allo-HSCT following complete remission (CR) achieved by CAR-T therapy versus by chemotherapy for B-cell acute lymphoblastic leukemia (B-ALL) has not been reported. We performed a parallel comparison of transplant outcomes in 105 consecutive B-ALL patients who received allo-HSCT after achieving CR with CAR-T therapy (n=27) or with chemotherapy (n=78). The CAR-T-allo-HSCT group had more patients in second CR compared to the chemotherapy-allo-HSCT group (78% vs. 37%; p

Neurofibrillary tangles composed of hyperphosphorylated tau and synaptic dysfunction are characteristics of Alzheimer’s disease (AD). However, the underlying molecular mechanisms remain poorly understood. Here, we identified Amphiphysin I mediates both tau phosphorylation and synaptic dysfunction in AD. Amphiphysin I is cleaved by a cysteine proteinase asparagine endopeptidase (AEP) at N278 in the brains of AD patients. The amount of AEP-generated N-terminal fragment of Amphiphysin I (1-278) is increased with aging. Amphiphysin I (1-278) inhibits clathrin-mediated endocytosis and induces synaptic dysfunction. Furthermore, Amphiphysin I (1-278) binds p35 and promotes its transition to p25, thus activates CDK5 and enhances tau hyperphosphorylation. Overexpression of Amphiphysin I (1-278) in the hippocampus of Tau P301S mice induces synaptic dysfunction, tau hyperphosphorylation, and cognitive deficits. However, overexpression of the N278A mutant Amphiphysin I, which resists the AEP-mediated cleavage, alleviates the pathological and behavioral defects. These findings suggest a mechanism of tau hyperphosphorylation and synaptic dysfunction in AD.

Objectives: To evaluate the performance of readout-segmented echo-planar imaging DWI (rs-EPI DWI) in detecting and characterizing breast cancers in a large Chinese cohort with comparison to dynamic contrast-enhanced MRI (DCE-MRI).Methods: The institutional review board approved this retrospective study with waived written informed consent. A total of 520 women (mean age, 43.1- ± 10.5-years) were included from July 2013 to October 2019. First, the ability of rs-EPI DWI in detecting breast lesions identified by DCE-MRI was evaluated. The lesion conspicuity of rs-EPI-DWI and DCE-MRI was compared using the Wilcoxon signed rank test. With pathology as a reference, the performance of rs-EPI DWI and DCE-MRI in distinguishing breast cancers was evaluated and compared using the Chi-square test.Results: Of 520 women, 327/520 (62.9%) patients had 423 lesions confirmed by pathology with 203 benign and 220 malignant lesions. The rs-EPI DWI can detect 90.8% (659/726) (reader 1) and 90.6% (663/732) (reader 2) of lesions identified by DCE-MRI. The lesion visibility was superior for DCE-MRI than rs-EPI-DWI (all p < 0.05). With pathology as a reference, the sensitivities and specificities of rs-EPI DWI in diagnosing breast cancers were 95.9% (211/220) and 85.7% (174/203) for reader 1 and 97.7% (215/220) and 86.2% (175/203) for reader 2. No significant differences were found for the performance of DCE-MRI and rs-EPI DWI in discriminating breast cancers (all p > 0.05).Conclusions: Although with an inferior lesion visibility, rs-EPI DWI can detect about 90% of breast lesions identified by DCE-MRI and has comparable diagnostic capacity to that of DCE-MRI in identifying breast cancer.

Objective: To present the evidence of the therapeutic effects and safety of Chinese herbal medicine (CHM) used with or without conventional western therapy for COVID-19.Methods: Clinical studies on the therapeutic effects and safety of CHM for COVID-19 were included. We summarized the general characteristics of included studies, evaluated methodological quality of randomized controlled trials (RCTs) using the Cochrane risk of bias tool, analyzed the use of CHM, used Revman 5.4 software to present the risk ratio (RR) or mean difference (MD) and their 95% confidence interval (CI) to estimate the therapeutic effects and safety of CHM.Results: A total of 58 clinical studies were identified including RCTs (17.24%, 10), non-randomized controlled trials (1.72%, 1), retrospective studies with a control group (18.97%, 11), case-series (20.69%, 12) and case-reports (41.38%, 24). No RCTs of high methodological quality were identified. The most frequently tested oral Chinese patent medicine, Chinese herbal medicine injection or prescribed herbal decoction were: Lianhua Qingwen granule/capsule, Xuebijing injection and Maxing Shigan Tang. In terms of aggravation rate, pooled analyses showed that there were statistical differences between the intervention group and the comparator group (RR 0.42, 95% CI 0.21 to 0.82, six RCTs; RR 0.38, 95% CI 0.23 to 0.64, five retrospective studies with a control group), that is, CHM plus conventional western therapy appeared better than conventional western therapy alone in reducing aggravation rate. In addition, compared with conventional western therapy, CHM plus conventional western therapy had potential advantages in increasing the recovery rate and shortening the duration of fever, cough and fatigue, improving the negative conversion rate of nucleic acid test, and increasing the improvement rate of chest CT manifestations and shortening the time from receiving the treatment to the beginning of chest CT manifestations improvement. For adverse events, pooled data showed that there were no statistical differences between the CHM and the control groups.Conclusion: Current low certainty evidence suggests that there maybe a tendency that CHM plus conventional western therapy is superior to conventional western therapy alone. The use of CHM did not increase the risk of adverse events.

Background: The World Health Organization characterized the Coronavirus disease 2019 (COVID-19) as a pandemic on March 11th. Many clinical trials on COVID-19 have been registered, and we aim to review the study characteristics and provide guidance for future trials to avoid duplicated effort. Methods: Studies on COVID-19 registered before March 3rd, 2020 on eight registry platforms worldwide were searched and the data of design, participants, interventions, and outcomes were extracted and analyzed. Results: Three hundred and ninety-three studies were identified and 380 (96.7%) were from mainland China, while 3 in Japan, 3 in France, 2 in the US, and 3 were international collaborative studies. Two hundred and sixty-six (67.7%) aimed at therapeutic effect, others were for prevention, diagnosis, prognosis, etc. Two hundred and two studies (51.4%) were randomized controlled trials. Two third of therapeutic studies tested Western medicines including antiviral drugs (17.7%), stem cell and cord blood therapy (10.2%), chloroquine and derivatives (8.3%), 16 (6.0%) on Chinese medicines, and 73 (27.4%) on integrated therapy of Western and Chinese medicines. Thirty-one studies among 266 therapeutic studies (11.7%) used mortality as primary outcome, while the most designed secondary outcomes were symptoms and signs (47.0%). Half of the studies (45.5%) had not started recruiting till March 3rd. Conclusion: Inappropriate outcome setting, delayed recruitment and insufficient numbers of new cases in China implied many studies may fail to complete. Strategies and protocols of the studies with robust and rapid data sharing are warranted for emergency public health events, helping the timely evidence-based decision-making.

Abstract Background The purpose of this study was to evaluate the efficacy of denosumab or zoledronic acid (ZA) using symptomatic skeletal events (SSEs) as the primary endpoint in Asian postmenopausal women with oestrogen-receptor-positive advanced breast cancer. Methods Asian postmenopausal women with oestrogen-receptor-positive advanced breast cancer receiving subcutaneous denosumab 120 mg Q4W, or intravenous ZA 4 mg Q4W until the primary analysis cut-off date were retrospectively analysed in the Hong Kong Practice-Based Cancer Research Center(HKCRC) from March 2011 to March 2013. The time to first on-study SSE that was assessed either clinically or through routine radiographic scans was the primary endpoint. Results 242 patients received denosumab or ZA treatment (n = 120, mean age of 64.9 years (SD 3.01) and n = 122, 65.4 years (3.44), respectively). The median times to first on-study SSE were 14.7 months (12.9–45.6) and 11.7 months (9.9–45.6) for denosumab and ZA, respectively (hazard ratio, HR 0.44, 95% CI 0.71–2.95; p = 0·0002). Compared with the ZA group, denosumab-treated patients had a significantly delayed time to first SSE (HR 0.65 [95% CI 0.29–1.45], p

Abstract Background δ-Tocotrienol is a naturally occurring proteasome inhibitor, which has the capacity to inhibit proliferation and induce apoptosis in several cancer cells obtained from several organs of humans, and other cancer cell lines. Moreover, results of plasma total mRNAs after δ-tocotrienol feeding to hepatitis C patients revealed significant inhibition in the expression of pro-inflammatory cytokines (TNF-α, VCAM1, proteasome subunits) and induction in the expression of ICAM1 and IFN-γ after post-treatment. This down-regulation of proteasome subunits leads to autophagy, apoptosis of immune cells and several genes. The present study describes RNA-sequence analysis of plasma total mRNAs obtained from δ-tocotrienol treatment of hepatitis C patients on gene expression regulated by proteasome. Methods Pooled specimens of plasma total mRNAs of pre-dose versus post-dose of δ-tocotrienol treatment of hepatitis C patients were submitted to RNA-sequence analyses. The data based on > 1 and 8-fold expression changes of 2136 genes were uploaded into “Ingenuity Pathway Analyses (IPA)” for core analysis, which describes possible canonical pathways, upstream regulators, diseases and functional metabolic networks. Results The IPA of “molecules” indicated fold change in gene expression of 953 molecules, which covered several categories of biological biomarkers. Out of these, gene expression of 220 related to present study, 12 were up-regulated, and 208 down-regulated after δ-tocotrienol treatment. The gene expression of transcription regulators (ceramide synthase 3 and Mohawk homeobox) were up-regulated, and gene expression of 208 molecules were down-regulated, involved in several biological functions (HSP90AB1, PSMC3, CYB5R4, NDUFB1, CYP2R1, TNFRF1B, VEGFA, GPR65, PIAS1, SFPQ, GPS2, EIF3F, GTPBP8, EIF4A1, HSPA14, TLR8, TUSSC2). IPA of “causal network” indicated gene regulators (676), in which 76 down-regulated (26 s proteasomes, interleukin cytokines, and PPAR-ligand-PPA-Retinoic acid-RXRα, PPARγ-ligand-PPARγ-Retinoic acid-RARα, IL-21, IL-23) with significant P-values. The IPA of “diseases and functions” regulators (85) were involved with cAMP, STAT2, 26S proteasome, CSF1, IFNγ, LDL, TGFA, and microRNA-155-5p, miR-223, miR-21-5p. The IPA of “upstream analysis” (934) showed 57 up-regulated (mainly 38 microRNAs) and 64 gene regulators were down-regulated (IL-2, IL-5, IL-6, IL-12, IL-13, IL-15, IL-17, IL-18, IL-21, IL-24, IL-27, IL-32), interferon β-1a, interferon γ, TNF-α, STAT2, NOX1, prostaglandin J2, NF-κB, 1κB, TCF3, and also miRNA-15, miRNA-124, miRNA-218-5P with significant activation of Z-Score (P

The aim of this study was to investigate the oxidative and proliferative effects of grape seed extract (GSE). Piglet intestinal epithelial cells (IPEC1) were selected as an unstressed cell model, or they were exposed to 400 μM H2O2 to establish a H2O2-stimulated cell model. The glutathione (GSH) and total antioxidant capacity in response to GSE addition were tested in the unstressed and H2O2-stimulated cell models. The relative mRNA levels of antioxidant or antioxidant enzymes and apoptosis-related genes were measured by Real-Time RT-PCR. In the unstressed status, the cell survival ratio and GSH increased with the addition of GSE at 1 and 10 μg/mL but diminished at 60 μg GSE/mL. The addition of 1 μg/mL GSE upregulated the mRNA expression levels of B-Cell Lymphoma protein-2 (Bcl-2), cysteine aspartases-3 (Caspase-3), cysteine aspartases-8 (Caspase-8) and glutathione peroxidase-1(GPx-1), while it downregulated that of Bcl2-associated X protein (Bax), copper-zinc superoxide dismutase, glutathione S-transferase (GST), thioredoxin, thioltransferase and thioredoxin reductase. As GSE reached 60 μg/mL, the tumor protein p53 (p53) and caspase-8 gene expressions were upregulated. In stressed status, 1 and 10 μg GSE/mL promoted the increase of GSH. H2O2- induced increases in Bax, p53, and Caspase-3 mRNA expressions were attenuated by the subsequent addition of 1 μg GSE/mL and promoted the gene expression of tumor necrosis factor-α, GPx-1 and thioltransferase (Ttas). Treatment with 60 μg GSE/ml resulted in a significant reduction in Bax, p53, manganese superoxide dismutase and GST mRNA expressions. These results indicate that GSE exhibits antioxidative and proliferative functions on unstressed IPEC1 cells at low and medium levels and oxidative and antiproliferative functions at high levels.

A long-term, high-resolution cropland dataset plays an essential part in accurately and systematically understanding the mechanisms that drive cropland change and its effect on biogeochemical processes. However, current widely used spatially explicit cropland databases are developed according to a simple downscaling model and are associated with low resolution. By combining historical county-level cropland archive data with natural and anthropogenic variables, we developed a random forest model to spatialize the cropland distribution in the Tuojiang River Basin (TRB) during 1911–2010, using a resolution of 30 m. The reconstruction results showed that the cropland in the TRB increased from 1.13 × 104 km2 in 1911 to 1.81 × 104 km2. In comparison with satellite-based data for 1980, the reconstructed dataset approximated the remotely sensed cropland distribution. Our cropland map could capture cropland distribution details better than three widely used public cropland datasets, due to its high spatial heterogeneity and improved spatial resolution. The most critical factors driving the distribution of TRB cropland include nearby-cropland, elevation, and climatic conditions. This newly reconstructed cropland dataset can be used for long-term, accurate regional ecological simulation, and future policymaking. This novel reconstruction approach has the potential to be applied to other land use and cover types via its flexible framework and modifiable parameters.

For thermal and loaded rock in engineering structures for some projects, triple-shear Drucker–Prager yield criteria, compaction coefficient K, damage variable correction factor δ, and thermal damage variable DT are introduced in a new thermomechanical (TM) constitutive model for the entire process. The compaction stage of rock in uniaxial compression test and the strain softening of rock caused by thermal attack are considered in this article. The damage evolution of rocks is described by a damage variable and a constitutive equation, which are in agreement with the actual thermal experimental breakage. The uniaxial compressive strength of granite subjected to a TM coupling effect can be predicted properly by this new unified constitutive model. The new TM unified constitutive model considering the compaction stage and post-failure stage is in good agreement with the test curves throughout the entire process. The coupling effect of heat and load in the total damage of rock has obvious nonlinear properties, but the coupling effect significantly weakens the specimens. By using the new TM unified constitutive model, the whole process of changes in rock damage with strain after high temperature can be calculated. Meanwhile, the model well represents the stress–strain curve at the post-failure stage. It is expected that this model can provide references for studying the mechanical response of the rock damage propagation characteristics in the future.

In order to investigate the effects of second-order hydrodynamic loads on a 15 MW floating offshore wind turbine (FOWT), this study employs a tool that integrates AQWA and OpenFAST to conduct fully coupled simulations of the FOWT subjected to wind and wave loadings. The load cases covering normal and extreme conditions are defined based on the met-ocean data observed at a specific site. The results indicate that the second-order wave excitations activate the surge mode of the platform. As a result, the surge motion is increased for each of the examined load case. In addition, the pitch, heave, and yaw motions are underestimated when neglecting the second-order hydrodynamics under the extreme condition. First-order wave excitation is the major contributor to the tower-base bending moments. The fatigue damage of the tower-base under the extreme condition is underestimated by 57.1% if the effect of second-order hydrodynamics is ignored. In addition, the accumulative fatigue damage over 25 years at the tower-base is overestimated by 16.92%. Therefore, it is suggested to consider the effects of second-order wave excitations of the floating platform for the design of the tower to reduce the cost of the FOWT.

In the study of rock mechanics, the variation of rock mechanical characteristics in high-temperature environments is always a major issue. The discrete element method and Voronoi modeling method were used to study the mechanical characteristics and crack evolution of granite specimens subjected to the high temperature and uniaxial compression test in order to study the internal crack evolution process of granite under the influence of high temperatures. Meanwhile, dependable findings were acquired when compared to experimental outcomes. A modified failure criterion was devised, and a Fish function was built to examine the evolution behavior of tensile and shear cracks during uniaxial compression, in order to better understand the evolution process of micro-cracks in granite specimens. Shear contacts occurred first, and the number of shear cracks reached its maximum value earliest, according to the findings. The number of tensile contacts then rapidly grew, whereas the number of shear cracks steadily declined. Furthermore, it was found that when temperature rises, the number of early tensile cracks grows. This study develops a fracture prediction system for rock engineering in high-temperature conditions.

In recent years, research on solar absorbers provides a significant breakthrough to solve the energy crisis. A perfect solar absorber based on a four-corner star array is designed and the absorption performance is analyzed numerically. The results show that the absorber reaches more than 90% of the full band in the range of 400–2000 nm. In particular, the absorption efficiency of the continuous more than 95% of the bandwidth reached 1391 nm, and the average absorption efficiency of the whole study band is more than 98%, and the loss of the solar spectrum only accounted for 2.7%. At the same time, the absorption efficiency can be adjusted by changing the geometric structure of the absorber. In addition, due to the perfect symmetry of the structure, it has an excellent insensitivity of the incident angle and polarization angle. In general, the proposed solar absorber has exciting prospects in solar energy collection and utilization, photothermal conversion and other related fields.

Objective This study aimed to evaluate the efficacy of beraprost sodium (BPS) or clopidogrel (CL) using vascular thromboembolic events (VTEs) of arteriovenous fistula as a primary endpoint in patients with end-stage renal disease (ESRD) undergoing arteriovenous fistula surgery. Methods We performed a multicentre, retrospective cohort study from August 2012 to August 2016. We studied patients with ESRD who underwent arteriovenous fistula surgery and received peroral administration of 40 µg BPS, three times per day, for 1 month, or 75 mg CL (initial dose of 300 mg), one time per day, for 1 month. The time to first on-study VTE was the primary endpoint. Results The BPS-treated cohort had a significantly delayed time to first VTE compared with the CL-treated cohort (hazard ratio 0.33, 95% confidence interval 0.18–0.56). An increased incidence of VTEs was detected in the 1-month follow-up, with rates of 2.4% and 8.7% for BPS and CL, respectively. This difference persisted over time, with rates of 8.0% and 18.1% at the final follow-up, respectively. Conclusion CL-treated patients with ESRD have a greater risk of VTEs compared with BPS-treated patients. CL-treated patients also tend to experience a VTE within the first month after cessation of oral administration.

Seed dormancy and germination are key events in plant development and are critical for crop production, and defects in seed germination or the inappropriate release of seed dormancy cause substantial losses in crop yields. Rice is the staple food for more than half of the world’s population, and preharvest sprouting (PHS) is one of the most severe problems in rice production, due to a low level of seed dormancy, especially under warm and damp conditions. Therefore, PHS leads to yield loss and a decrease in rice quality and vitality. We reveal that mutation of OsbZIP09 inhibited rice PHS. Analysis of the expression of OsbZIP09 and its encoded protein sequence and structure indicated that OsbZIP09 is a typical bZIP transcription factor that contains conserved bZIP domains, and its expression is induced by ABA. Moreover, RNA sequencing (RNA-seq) and DNA affinity purification sequencing (DAP-seq) analyses were performed and 52 key direct targets of OsbZIP09 were identified, including OsLOX2 and Late Embryogenesis Abundant (LEA) family genes, which are involved in controlling seed germination. Most of these key targets showed consistent changes in expression in response to abscisic acid (ABA) treatment and OsbZIP09 mutation. The data characterize a number of key target genes that are directly regulated by OsbZIP09 and contribute to revealing the molecular mechanism that underlies how OsbZIP09 controls rice seed germination.

Pile foundation is one of the common foundation forms in marine geotechnical engineering, especially in wind power engineering. Its operation safety is seriously affected by many uncertainties, such as the randomness of ground motion in intensity and frequency. The stochastic reliability analysis method can better characterize these uncertainties in the evaluation of the safety performance of pile foundation. The probability density functions (PDFs) of stochastic systems are important prerequisites for reliability analysis. However, for geotechnical problems, the coupling between parametric and excitation randomness and the nonlinear mechanical properties of rock and soil make it very difficult to obtain the associated PDFs. Instead, the probability density evolution method (PDEM) is introduced and is used to investigate the static and dynamic reliability of laterally loaded piles as an example of a geotechnical problem. Compared with Monte Carlo stochastic simulations, PDEM-based computing is shown to be highly efficient when applied to the seismic design of pile in geotechnical engineering, and its calculation efficiency is 20 times of the former for the seismic dynamic reliability of pile foundation. This study provides a new reference for the efficient design and safety evaluation of offshore pile foundation engineering based on static and dynamic reliability of multiple random factors.

Boson sampling is a promising candidate for quantum supremacy. It requires to sample from a complicated distribution, and is trusted to be intractable on classical computers. Among the various classical sampling methods, the Markov chain Monte Carlo method is an important approach to the simulation and validation of boson sampling. This method however suffers from the severe sample loss issue caused by the autocorrelation of the sample sequence. Addressing this, we propose the sample caching Markov chain Monte Carlo method that eliminates the correlations among the samples, and prevents the sample loss at the meantime, allowing more efficient simulation of boson sampling. Moreover, our method can be used as a general sampling framework that can benefit a wide range of sampling tasks, and is particularly suitable for applications where a large number of samples are taken.

A set of 899 L. gmelinii expression sequence tags (ESTs), available at the National Center of Biotechnology Information (NCBI), was employed to address the feasibility on development of simple sequence repeat (SSR) markers for Larch species. Totally, 634 non-redundant unigenes including 145 contigs and 489 singletons were finally identified and mainly involved in biosynthetic, metabolic processes and response to stress according to BLASTX results, gene ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) maps. Approximately 11.7% (74) unigenes contained 90 candidate SSRs, which were mainly trinucleotides (29, 32.2%) and dinucleotides (26, 28.9%). A relatively high frequency of SSRs was respectively found in the Open Reading Frame (ORF, about 54.4%) and 5′-untranslated region (5′-UTR, 31.2%), while a low frequency was observed in the 3′-untranslated region (3′-UTR, about 14.4%). Of the 45 novel EST-SSRs markers, nine were found to be polymorphic at two L. gmelinii populations. The number of alleles per locus (Na) ranged from two to four, and the observed (Ho) and expected (He) heterozygosity values were 0.200–0.733 and 0.408–0.604, respectively. The inbreeding coefficients (FIS) for all loci were more than zero except Lg41. Most of these 9EST-SSR markers were transferable to its related species L. kaempferi, L. principis-rupprechtii and L. olgensis. These novel EST-SSRs will be useful for further research on comparative genomics, genetic resources conservation and molecular breeding in larch trees.

Human parvovirus B19 (B19V) infection of primary human erythroid progenitor cells (EPCs) arrests infected cells at both late S-phase and G2-phase, which contain 4N DNA. B19V infection induces a DNA damage response (DDR) that facilitates viral DNA replication but is dispensable for cell cycle arrest at G2-phase; however, a putative C-terminal transactivation domain (TAD2) within NS1 is responsible for G2-phase arrest. To fully understand the mechanism underlying B19V NS1-induced G2-phase arrest, we established two doxycycline-inducible B19V-permissive UT7/Epo-S1 cell lines that express NS1 or NS1mTAD2, and examined the function of the TAD2 domain during G2-phase arrest. The results confirm that the NS1 TAD2 domain plays a pivotal role in NS1-induced G2-phase arrest. Mechanistically, NS1 transactivated cellular gene expression through the TAD2 domain, which was itself responsible for ATR (ataxia-telangiectasia mutated and Rad3-related) activation. Activated ATR phosphorylated CDC25C at serine 216, which in turn inactivated the cyclin B/CDK1 complex without affecting nuclear import of the complex. Importantly, we found that the ATR-CHK1-CDC25C-CDK1 pathway was activated during B19V infection of EPCs, and that ATR activation played an important role in B19V infection-induced G2-phase arrest.

Due to the diverse medicinal effects, polyphenols are among the most intensively studied natural products. However, it is a great challenge to elucidate the polypharmacological mechanisms of polyphenols. To address this challenge, we establish a method for identifying multiple targets of chemical agents through analyzing the module profiles of gene expression upon chemical treatments. By using FABIA algorithm, we have performed a biclustering analysis of gene expression profiles derived from Connectivity Map (cMap), and clustered the profiles into 49 gene modules. This allowed us to define a 49 dimensional binary vector to characterize the gene module profiles, by which we can compare the expression profiles for each pair of chemical agents with Tanimoto coefficient. For the agent pairs with similar gene expression profiles, we can predict the target of one agent from the other. Drug target enrichment analysis indicated that this method is efficient to predict the multiple targets of chemical agents. By using this method, we identify 148 targets for 20 polyphenols derived from cMap. A large part of the targets are validated by experimental observations. The results show that the medicinal effects of polyphenols are far beyond their well-known antioxidant activities. This method is also applicable to dissect the polypharmacology of other natural products.

Human bocavirus 1 (HBoV1) infects well-differentiated (polarized) human airway epithelium (HAE) cultured at an air-liquid interface (ALI). In the present study, we applied next-generation RNA sequencing to investigate the genome-wide transcription profile of HBoV1, including viral mRNA and small RNA transcripts, in HBoV1-infected HAE cells. We identified novel transcription start and termination sites and confirmed the previously identified splicing events. Importantly, an additional proximal polyadenylation site (pA)p2 and a new distal polyadenylation site (pA)dREH lying on the right-hand hairpin (REH) of the HBoV1 genome were identified in processing viral pre-mRNA. Of note, all viral nonstructural proteins-encoding mRNA transcripts use both the proximal polyadenylation sites [(pA)p1 and (pA)p2] and distal polyadenylation sites [(pA)d1 and (pA)dREH] for termination. However, capsid proteins-encoding transcripts only use the distal polyadenylation sites. While the (pA)p1 and (pA)p2 sites were utilized at roughly equal efficiency for proximal polyadenylation of HBoV1 mRNA transcripts, the (pA)d1 site was more preferred for distal polyadenylation. Additionally, small RNA-seq analysis confirmed there is only one viral noncoding RNA (BocaSR) transcribed from nt 5199–5340 of the HBoV1 genome. Thus, our study provides a systematic and unbiased transcription profile, including both mRNA and small RNA transcripts, of HBoV1 in HBoV1-infected HAE-ALI cultures.

Colored calla lily is the short name for the species or hybrids in section Aestivae of genus Zantedeschia. It is currently one of the most popular flower plants in the world due to its beautiful flower spathe and long postharvest life. However, little genomic information and few molecular markers are available for its genetic improvement. Here, de novo transcriptome sequencing was performed to produce large transcript sequences for Z. rehmannii cv. ‘Rehmannii’ using an Illumina HiSeq 2000 instrument. More than 59.9 million cDNA sequence reads were obtained and assembled into 39,298 unigenes with an average length of 1,038 bp. Among these, 21,077 unigenes showed significant similarity to protein sequences in the non-redundant protein database (Nr) and in the Swiss-Prot, Gene Ontology (GO), Cluster of Orthologous Group (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Moreover, a total of 117 unique transcripts were then defined that might regulate the flower spathe development of colored calla lily. Additionally, 9,933 simple sequence repeats (SSRs) and 7,162 single nucleotide polymorphisms (SNPs) were identified as putative molecular markers. High-quality primers for 200 SSR loci were designed and selected, of which 58 amplified reproducible amplicons were polymorphic among 21 accessions of colored calla lily. The sequence information and molecular markers in the present study will provide valuable resources for genetic diversity analysis, germplasm characterization and marker-assisted selection in the genus Zantedeschia.

Many studies aimed at investigating bone repair have been conducted through animal models in recent years. However, limitations do exist in these models due to varying regeneration potential among different animal species. Even using the same animal, big differences exist in the size of critical size defects (CSD) involving the same region. This study aimed to investigate the standardization of radial bone defect models in rabbits and further establish more reliable CSD data. A total of 40 6-month-old New Zealand white rabbits of clean grade totaling 80 radial bones were prepared for bone defect models, according to the principle of randomization. Five different sizes (1.0, 1.2, 1.4, 1.7 and 2.0 cm) of complete periosteal defects were introduced under anesthesia. At 12 weeks postoperatively, with the gradual increase in defect size, the grades of bone growth were significantly decreased in all 5 groups. X-ray, CT scans and H&E staining of the 1.4, 1.7, and 2.0-cm groups showed lower grades of bone growth than that of the 1.0 and 1.2-cm groups respectively (P < 0.05). Using rabbit radial defect model involving 6-month-old healthy New Zealand white rabbits, this study indicates that in order to be critical sized, defects must be greater than 1.4 cm.

Seed germination, a pivotal process in higher plants, is precisely regulated by various external and internal stimuli, including brassinosteroid (BR) and gibberellin (GA) phytohormones. The molecular mechanisms of crosstalk between BRs and GAs in regulating plant growth are well established. However, whether BRs interact with GAs to coordinate seed germination remains unknown, as do their common downstream targets. In the present study, 45 differentially expressed proteins responding to both BR and GA deficiency were identified using isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis during seed germination. The results indicate that crosstalk between BRs and GAs participates in seed germination, at least in part, by modulating the same set of responsive proteins. Moreover, most targets exhibited concordant changes in response to BR and GA deficiency, and gene ontology (GO) indicated that most possess catalytic activity and are involved in various metabolic processes. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis was used to construct a regulatory network of downstream proteins mediating BR- and GA-regulated seed germination. The mutation of GRP, one representative target, notably suppressed seed germination. Our findings not only provide critical clues for validating BR⁻GA crosstalk during rice seed germination, but also help to optimise molecular regulatory networks.

Although 109 WRKY genes have been identified in the rice genome, the functions of most are unknown. Here, we show that OsWRKY13 plays a pivotal role in rice disease resistance. Overexpression of OsWRKY13 can enhance rice resistance to bacterial blight and fungal blast, two of the most devastating diseases of rice worldwide, at both the seedling and adult stages, and shows no influence on the fertility. This overexpression was accompanied by the activation of salicylic acid (SA) synthesis-related genes and SA-responsive genes and the suppression of jasmonic acid (JA) synthesis-related genes and JA-responsive genes. OsWRKY13 bound to the promoters of its own and at least three other genes in SA- and JA-dependent signaling pathways. Its DNA-binding activity was influenced by pathogen infection. These results suggest that OsWRKY13, as an activator of the SA-dependent pathway and a suppressor of JA-dependent pathways, mediates rice resistance by directly or indirectly regulating the expression of a subset of genes acting both upstream and downstream of SA and JA. Furthermore, OsWRKY13 will provide a transgenic tool for engineering wider-spectrum and whole-growth-stage resistance rice in breeding programs.

The resistivity of oil impregnated paper will decrease during its aging process. This paper takes paper resistivity as an assessment index to evaluate the insulation condition of oil impregnated paper in power transformer. The feasibility of this method are discussed in two aspects: reliability and sensitivity. Iterative inversion of paper resistivity was combined with finite element simulation. Both the bisection method and Newton’s method were used as iterative methods. After the analysis and comparison, Newton’s method was selected as the first option of paper resistivity iteration for its faster convergence. In order to consider the spatial distribution characteristic of paper aging and enhance the calculation accuracy, the resistivity calculation is expanded to a multivariate iteration based on Newton’s method, in order to consider the spatial distribution characteristic of paper aging and improve the calculation accuracy. This paper presents an exploratory research on condition assessment of oil impregnated paper insulation, and provides some reference to the security and economy operation of power transformers.

As a special class of short non-coding RNAs, microRNAs (a.k.a. miRNAs or miRs) have been reported to perform important roles in various biological processes by regulating respective target genes. However, significant barriers exist during biologists' conventional miR knowledge discovery. Emerging semantic technologies, which are based upon domain ontologies, can render critical assistance to this problem. Our previous research has investigated the construction of a miR ontology, named Ontology for MIcroRNA Target Prediction (OMIT), the very first of its kind that formally encodes miR domain knowledge. Although it is unavoidable to have a manual component contributed by domain experts when building ontologies, many challenges have been identified for a completely manual development process. The most significant issue is that a manual development process is very labor-intensive and thus extremely expensive. Therefore, we propose in this paper an innovative ontology development methodology. Our contributions can be summarized as: (i) We have continued the development and critical improvement of OMIT, solidly based on our previous research outcomes. (ii) We have explored effective and efficient algorithms with which the ontology development can be seamlessly combined with machine intelligence and be accomplished in a semi-automated manner, thus significantly reducing large amounts of human efforts. A set of experiments have been conducted to thoroughly evaluate our proposed methodology.

Legumain-based DNA vaccines have potential to protect against breast cancer. However, the lack of a safe and efficient oral delivery system restricts its clinical application. Here, we constructed alginic acid-coated chitosan nanoparticles (A.C.NPs) as an oral delivery carrier for a legumain DNA vaccine. First, we tested its characteristic in acidic environments in vitro. DNA agarose electrophoresis data show that A.C.NPs protected DNA better from degradation in acidic solution (pH 1.5) than did chitosan nanoparticles (C.NPs). Furthermore, size distribution analysis showed that A.C.NPs tended to aggregate and form micrometer scale complexes in pH

The gene editing and synthetic biological tools have led to the implementation of diverse metabolic engineering approaches to enhance the production of specific enzymes. Microbial keratinases can convert keratin wastes into valuable compounds for mankind. Since the market for keratinases cannot be satisfied by production from wild hosts, it is obligatory to develop hosts with high keratinase yields. The intention of this review is to evaluate microbial keratinase advancement through protein engineering, breeding techniques, and fermentation optimization. The main aim of protein engineering is to improve the heat resistance ability and catalytic activity of keratinases by employing mutagenesis methods. Moreover, modifying the expression elements and host engineering are also two unique ways to augment the keratinase yield. Intending to accelerate the production of modified keratinase, this review attempts to highlight the optimization of expression elements, such as promoter engineering, UTR, signal peptide, and codon optimization. Moreover, the approaches of host engineering including strengthening precursor supply, membrane surface engineering, and optimization of secretion pathways were also explained here. Furthermore, it is also essential to optimize the medium composition and fermentation condition for high keratinase yield. This review also addressed the present advancements, difficulties, and tendencies in the field of microbial keratinase production, along with its potential. [ABSTRACT FROM AUTHOR]