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Early-onset epilepsy following ischemic stroke is a severe neurological condition, the pathogenesis of which remains incompletely understood. Recent studies suggest that Neural stem/progenitor cells (NSPCs) play a crucial role in the disease process, yet the precise molecular mechanisms regulating NSPCs have not been thoroughly investigated. This study utilized single-cell transcriptome sequencing and bioinformatics analysis to identify disease-related genes, which were subsequently validated in both in vitro and in vivo experiments. The findings revealed that Hsp90aa1 (heat shock protein 90 kDa alpha, class A member 1), Jun proto-oncogene (JUN), and CC Motif Ligation 2 (Ccl2) constitute an important regulatory axis influencing the migration and differentiation of NSPCs, potentially impacting the onset and progression of early-onset epilepsy post-ischemic stroke. Additionally, the expression of Hsp90aa1 was found to influence the likelihood of seizure occurrence and the severity of brain ischemia.

Magnetite (Fe3O4) particles have been widely reported to enhance the anammox's activity in anammox granular sludge (AnGS), yet the underlying mechanisms remain unclear. This study demonstrates that both Fe3O4 microparticles (MPs) and nanoparticles (NPs) at a dosage of 200 mg Fe3O4/L significantly increased the specific anammox activity (SAA) of AnGS. Additionally, the transcriptional activities of the hzs and hdh genes involved in the anammox process, as well as the heme c content in AnGS, were also notably enhanced. Notably, Fe3O4 NPs were more effective than MPs in boosting anammox activity within AnGS. Mechanistically, Fe3O4 MPs released free iron, which anammox bacteria utilized to promote the synthesis of key enzymes, thereby enhancing their activity. Compared to MPs, Fe3O4 NPs not only elevated the synthesis of these key enzymes to a higher level but also induced a nanofluids effect on the surface of AnGS, improving substrate permeability and accessibility to intragranular anammox bacteria. Moreover, the nanofluids effect was identified as the primary mechanism through which Fe3O4 NPs enhanced anammox activity within AnGS. These findings provide new insights into the effects of nanoparticles on granular sludge systems, extending beyond AnGS.

Summary: Although more and more evidence has supported that metabolic syndrome (MS) is linked to ischemic stroke (IS), the molecular mechanism and genetic association between them has not been investigated. Here, we combined the existing single-cell RNA sequencing (scRNA-seq) data and mendelian randomization (MR) for stroke to understand the role of dysregulated metabolism in stroke. The shared hub genes were identified with machine learning and WGCNA. A total of six upregulated DEGs and five downregulated genes were selected for subsequent analyses. Nine genes were finally identified with random forest, Lasso regression, and XGBoost method as a potential diagnostic model. scRNA-seq also show the abnormal glycolysis level in most cell clusters in stroke and associated with the expression level of hub genes. The genetic relationship between IS and MS was verified with MR analysis. Our study reveals the common molecular profile and genetic association between ischemic stroke and metabolic syndrome.

Summary: The medial entorhinal cortex (MEC) is crucial for contextual memory, yet its role in context-induced retrieval of morphine withdrawal memory remains unclear. This study investigated the role of the MEC and its projection neurons from MEC layer 5 to the basolateral amygdala (BLA) (MEC−BLA neurons) in context-induced retrieval of morphine withdrawal memory. Results show that context activates the MEC in morphine withdrawal mice, and the inactivation of the MEC inhibits context-induced retrieval of morphine withdrawal memory. At neural circuits, context activates MEC−BLA neurons in morphine withdrawal mice, and the inactivation of MEC−BLA neurons inhibits context-induced retrieval of morphine withdrawal memory. But MEC−BLA neurons are not activated by conditioning of context and morphine withdrawal, and the inhibition of MEC−BLA neurons do not influence the coupling of context and morphine withdrawal memory. These results suggest that MEC−BLA neurons are critical for the retrieval, but not for the formation, of morphine withdrawal memory.

Methane metabolism, driven by methanogenic and methanotrophic microorganisms, plays a pivotal role in the carbon cycle. As seawater intrusion and soil salinization rise due to global environmental shifts, understanding how salinity affects methane emissions, especially in deep strata, becomes imperative. Yet, insights into stratigraphic methane release under varying salinity conditions remain sparse. Here we investigate the effects of salinity on methane metabolism across terrestrial and coastal strata (15–40 m depth) through in situ and microcosm simulation studies. Coastal strata, exhibiting a salinity level five times greater than terrestrial strata, manifested a 12.05% decrease in total methane production, but a staggering 687.34% surge in methane oxidation, culminating in 146.31% diminished methane emissions. Salinity emerged as a significant factor shaping the methane-metabolizing microbial community's dynamics, impacting the methanogenic archaeal, methanotrophic archaeal, and methanotrophic bacterial communities by 16.53%, 27.25%, and 22.94%, respectively. Furthermore, microbial interactions influenced strata system methane metabolism. Metabolic pathway analyses suggested Atribacteria JS1's potential role in organic matter decomposition, facilitating methane production via Methanofastidiosales. This study thus offers a comprehensive lens to comprehend stratigraphic methane emission dynamics and the overarching factors modulating them.

Summary: Both therapeutic hypothermia and neural stem cells (NSCs) transplantation have shown promise in neuroprotection and neural repair after brain injury. However, the effects of therapeutic hypothermia on neuronal differentiation of NSCs are not elucidated. In this study, we aimed to investigate whether mild hypothermia promoted neuronal differentiation in cultured and transplanted human NSCs (hNSCs). A significant increase in neuronal differentiation rate of hNSCs was found when exposed to 35°C, from 33% to 45% in vitro and from 7% to 15% in vivo. Additionally, single-cell RNA sequencing identified upregulation of RNA-binding motif protein 3 (RBM3) in neuroblast at 35°C, which stabilized the SRY-box transcription factor 11 (SOX11) mRNA and increased its protein expression, leading to an increase in neuronal differentiation of hNSCs. In conclusion, our study highlights that mild hypothermia at 35°C enhances hNSCs-induced neurogenesis through the novel RBM3-SOX11 signaling pathway, and provides a potential treatment strategy in brain disorders.

Watermelon is an important horticultural plant. A bibliometric analysis of the watermelon literature was carried out in order to analyze the research state, hotspots, and trends, as well as to highlight the overall watermelon research development from a holistic viewpoint. The summary of watermelon research is given via metrological analysis based on a set of indices using a newly built Bibliometrix R-package tool. This study gathered 6,632 documents indexed in the Core Collection of Web of Science (WoS) in the domain of watermelon from 1992 to 2022 using bibliometrix. The results indicated that the number of published articles showed an apparently upward trend. The United States was in the first place, with Plant Disease being the most productive journal. Levi A from the United States Department of Agriculture-Agricultural Research Service is the most prolific author, and Levi A is the most cited; The most frequently used keywords by authors are ''growth'', ''resistance'', ''identification'', ''yield'', ''quality'' ''plants'', ''watermelon stomach'' and ''expression''; The most talked-about issues in this subject are resistance, yield, and quality, which highlight the crucial research areas. To effectively comprehend the turning moments for future research, it is useful to monitor the hotspots and frontiers of watermelon studies. The results highlight the future paths for study in the field of watermelon and provide useful information for researchers interested in the topic.

Introduction and Objectives: Cholangiocarcinoma (CCA) is characterized by early distant invasion and metastasis, whereas the underlying mechanism is still obscure. Increasing evidence shows that collagen type Ι alpha 1 (COL1A1) is a gene associated with the progression of multiple diseases. Here, we attempted to investigate the role of COL1A1 in CCA. Materials and Methods: The expression of COL1A1 between tumor tissues and adjacent normal tissues obtained from CCA patients was detected by Western blot and immunofluorescence, followed by analysis of its clinical significance. Then, the biological effects of COL1A1 overexpression or knockdown on CCA cells were evaluated in vitro and in vivo. Finally, molecular mechanism of COL1A1 in regulating the invasion and metastasis of CCA cells was determined by a series of experiments. Results: COL1A1 expression was significantly higher in CCA pathological tissues than in corresponding adjacent normal tissues. Analysis of 83 CCA patients showed that higher expression of COL1A1 was correlated with poorer patient prognosis. Notably, overexpression or knockdown experiments revealed that COL1A1 contributed to the migration and invasion, as well as epithelial-to-mesenchymal transition (EMT), in CCA cells. Further investigations demonstrated that matrix metalloproteinase-2 (MMP2) promoted COL1A1 upregulation via the integrin alpha Ⅴ pathway, therefore affecting ECM remodelling and inducing EMT in CCA cells. Moreover, COL1A1 expression was positively related to PD-1 and PD-L1 in CCA, and COL1A1 increased PD-L1 expression by activating the NF-κB pathway. Conclusions: COL1A1 plays an important role in regulating CCA progression and may act as a promising biomarker and therapeutic target for CCA.

Background: During endoscopic submucosal dissection (ESD) for gastric lesions with fibrosis, appropriate traction could provide clear submucosal dissection visualization to improve safety and efficiency of procedures. Therefore, the aim of this study was to evaluate the feasibility of magnetic ring-assisted ESD (MRA-ESD) for gastric fibrotic lesions. Method: In the eight healthy beagles, 2–3 mL of 50% glucose solution was injected into submucosal layer of the stomach to induce gastric fibrotic lesions. A week after submucosal injection, two endoscopists at different levels performed MRA-ESD or standard ESD (S-ESD) for gastric simulated lesions, respectively. The magnetic traction system consisted of external handheld magnet and internal magnetic ring. The feasibility and procedure outcomes of the magnetic traction system were mainly evaluated. Results: Forty-eight gastric simulated lesions with ulceration were confirmed to have submucosal fibrosis formation by preoperative endoscopic ultrasonography. The magnetic traction system could be easily established, only took 1.57 min, and allowed excellent submucosal visualization. The total procedure time was significantly shorter in the MRA-ESD group than in the S-ESD group for both endoscopists (mean: 46.83 vs. 25.09 min, p

Due to principle constraints of the conventional transverse flux permanent magnet linear machine, its PM utilization is less than half. Besides, the complex structure and the serious magnetic leakage also lead to the limited thrust increase and the sharp decline of power factor. Therefore, the high-power-factor transverse-flux permanent-magnet linear machine (HPF-TFPMLM) using the concentrated-flux teeth is proposed in this paper. First, the topology and operation principle of the circumferentially distributed HPF-TFPMLM are investigated. The concentrated-flux teeth and multi-unit concentrated windings are introduced to improve power factor of the HPF-TFPMLM. To further study the proposed HPF-TFPMLM, its counterpart of the axial distributed topology is proposed for comparative analysis. Then, the key electromagnetic performance of power factor, thrust and thrust density of the HPF-TFPMLM are analyzed. Its performances including no-load EMF, magnetic field distribution and overload capacity are evaluated and compared with its counterpart. Finally, the feasibility and correctness of the investigated HPF-TFPMLM are verified.

Summary: Background: Chronic cough is a common respiratory symptom with an impact on daily activities and quality of life. Global prevalence data are scarce and derive mainly from European and Asian countries and studies with outcomes other than chronic cough. In this study, we aimed to estimate the prevalence of chronic cough across a large number of study sites as well as to identify its main risk factors using a standardised protocol and definition. Methods: We analysed cross-sectional data from 33,983 adults (≥40 years), recruited between Jan 2, 2003 and Dec 26, 2016, in 41 sites (34 countries) from the Burden of Obstructive Lung Disease (BOLD) study. We estimated the prevalence of chronic cough for each site accounting for sampling design. To identify risk factors, we conducted multivariable logistic regression analysis within each site and then pooled estimates using random-effects meta-analysis. We also calculated the population attributable risk (PAR) associated with each of the identifed risk factors. Findings: The prevalence of chronic cough varied from 3% in India (rural Pune) to 24% in the United States of America (Lexington,KY). Chronic cough was more common among females, both current and passive smokers, those working in a dusty job, those with a history of tuberculosis, those who were obese, those with a low level of education and those with hypertension or airflow limitation. The most influential risk factors were current smoking and working in a dusty job. Interpretation: Our findings suggested that the prevalence of chronic cough varies widely across sites in different world regions. Cigarette smoking and exposure to dust in the workplace are its major risk factors. Funding: Wellcome Trust.

Long-term high sugar accelerates skin ageing via glycosylation. In this study, microalgae-derived active compounds were investigated for their potentially beneficial effects on skin ageing. Two carotenoids, namely astaxanthin (in the form of either monoester or diester) and lutein, were separated from the green alga Haematococcus pluvialis. These carotenoid fractions significantly attenuated the high glucose (HG)-induced cell injury and oxidative stress. They also inhibited the formation of Nε-Carboxymethyllysine (CML), a typical product of glycosylation. The degradation of collagen I and III caused by HG was substantially recovered, in which astaxanthin monoester exhibited the most prominent effect. This was attributed to the down-regulation of matrix metalloproteinase (MMP)-2 mRNA expression. Through proteomics analyses, six hub proteins were identified, which work as highly connected nodes in the protective effects of astaxanthin monoester against HG stress. These findings strongly suggest the benefits of microalgae-derived carotenoids in alleviating HG-induced skin ageing.

Clinically, neuropathic pain treatment remains a challenging issue because the major therapy, centred around pharmacological intervention, is not satisfactory enough to patient by reason of low effectiveness and more adverse reaction. Therefore, it is still necessary to find more effective and safe therapy to ameliorate neuropathic pain. The purpose of this study was to explore the antinociceptive effect of Echinacoside (ECH), an active compound of Cistanche deserticola Ma, on peripheral neuropathic pain induced by chronic constriction injury (CCI) in mice, and to demonstrate its potential mechanism in vivo and vitro. In the present study, results showed that intraperitoneal administration of ECH (50, 100, and 200 mg/kg) could alleviate mechanical allodynia, cold allodynia and thermal hyperalgesia via behavioural test. Moreover, the structure and function of injured sciatic nerve by CCI were taken a turn for the better to a certain extent after ECH treatment using histopathological and electrophysiological test. Furthermore, ECH repressed the expression of the P2X7R and FKN and reduced the expression and release of the IL-1β, IL-6 and TNF-α. Besides, ECH could decrease Ca2+ influx and Cats efflux and inhibit phosphorylation of p38MAPK. To sum up, the present study illustrated that ECH could alleviate peripheral neuropathic pain by inhibiting microglia overactivation and inflammation through P2X7R/FKN/CX3CR1 signalling pathway in spinal cord. This study would provide a new perspective and strategy for the pharmacological treatment on neuropathic pain.

The objective of this study was to investigate the effects of dietary L-theanine (LTH) supplementation on finishing pigs' antioxidant capacity and lipid metabolism. Results showed that dietary LTH supplementation increased the activity of antioxidant enzymes and decreased the content of malondialdehyde in serum, longissimus dorsi muscle (LD muscle), and liver of finishing pigs (P

The DEPDC5 gene (OMIM 614191) has been proven to be a frequent cause of familial and sporadic focal epilepsy. A human induced pluripotent stem cell (iPSC) line was generated from a child diagnosed with focal epilepsy, which was caused by DEDPC5 mutation. The iPSC line expresses high pluripotency markers, carries the DEDPC5 mutation, and can differentiate into three germ layers in vitro. The iPSC lines offer a promising technique for studying the pathogenesis and conducting drug screening of DEDPC5-related epilepsy.

Deciphering gene function is fundamental to engineering of microbiology. The clustered regularly interspaced short palindromic repeats (CRISPR) system has been adapted for gene repression across a range of hosts, creating a versatile tool called CRISPR interference (CRISPRi) that enables genome-scale analysis of gene function. This approach has yielded significant advances in the design of genome-scale CRISPRi libraries, as well as in applications of CRISPRi screening in medical and industrial microbiology. This review provides an overview of the recent progress made in pooled and arrayed CRISPRi screening in microorganisms and highlights representative studies that have employed this method. Additionally, the challenges associated with CRISPRi screening are discussed, and potential solutions for optimizing this strategy are proposed.

For the demand of high reliability in the aerospace field, multi-phase electric machines have become a research hotspot for the fault-tolerant ability. Inter-turn short-circuit fault is one of the main fault forms in the electric machine. The fault mechanism, diagnosis method and fault-tolerant control strategy of inter-turn short-circuit fault of the open-winding five-phase permanent-magnet synchronous machine that is used as the starter/generator system are studied in this paper. Firstly, the fault mechanism of the inter-turn short-circuit fault (ITSCF) is analyzed. The characteristics of phase voltage, inter-turn short-circuit current and torque ripple are compared using the phase winding open-circuit method and the phase winding short-circuit method. Secondly, a fault diagnosis method based on the information fusion is proposed, which determines the fault phase firstly and then identifies the inter-turn short-circuit ratio. According to the fault mechanism, a fault-tolerant control method is proposed, in which the accurate inter-turn short-circuit ratio and inter-turn short-circuit current are not required. The proposed fault-tolerant control method has similar torque ripple compared with the control method using accurate inter-turn short-circuit ratio and inter-turn short-circuit current. Finally, the effectiveness of the proposed fault-tolerant control method has been verified by experimental results.

This study was conducted to investigate the effects of betaine on lipid metabolism and m6A RNA methylation in finishing pigs fed a low-energy diet. The results showed that betaine significantly enhanced serum concentration of triglyceride and reduced the relative weight of flare fat (P

ABSTRACT: Yucca schidigera extract (YSE) is a green feed additive that is known to reduce toxic gas emissions and promote intestinal health in animal production. This study investigated the potential of dietary YSE supplementation to mitigate the negative effect of Clostridium perfringens and coccidia infection on productive performance and gut health in laying hens. A total of 48 Lohmann gray laying hens (35 wk of age) were randomly allotted to 1 of 2 groups (n = 24) fed with either a basal diet or a YSE-supplemented diet for 45 d. From d 36 to 45, half of the hens in each group were orally administrated with Clostridium perfringens type A and coccidia. This challenge impaired productive performance and egg quality (P < 0.05), destroyed jejunal morphology and functions (P < 0.05), induced jejunal epithelial cell apoptosis (P < 0.05), and downregulated the antioxidant capacity and Nrf2 pathway expression of jejunal mucosa (P < 0.05) in laying hens. Supplementing YSE in the laying hen diet, to some extents, improved productive performance and egg quality (P < 0.05), and alleviated the effect of challenge on morphology, functions, cell apoptosis, and antioxidant capacity in the jejunum (P < 0.05). Overall, the results suggested that dietary YSE supplementation might mitigate the negative effects of Clostridium perfringens and coccidia infection on gut health, and thereby improve the productive performance and egg quality of laying hens, possibly through enhancing the antioxidant capacity of the jejunum.

Summary: Neuraminidase is suggested as an important component for developing a universal influenza vaccine. Targeted induction of neuraminidase-specific broadly protective antibodies by vaccinations is challenging. To overcome this, we rationally select the highly conserved peptides from the consensus amino acid sequence of the globular head domains of neuraminidase. Inspired by the B cell receptor evolution process, a reliable sequential immunization regimen is designed to result in immuno-focusing by steering bulk immune responses to a selected region where broadly protective B lymphocyte epitopes reside. After priming neuraminidase protein-specific antibody responses in C57BL/6 or BALB/c inbred mice strains by immunization or pre-infection, boost immunizations with certain neuraminidase-derived peptide-keyhole limpet hemocyanin conjugates significantly strengthened serum neuraminidase inhibition activities and cross-protections. Overall, this study provides proof of concept for a peptide-based sequential immunization strategy for achieving targeted induction of cross-protective antibody response, which provides references for designing universal vaccines against other highly variable pathogens.

Dietary fiber (DF) has long been looked as an essential “nutrients” both for animals and humans as it can promote the intestinal tract development and modulate the intestinal epithelium functions and the gut microbiota. This study was conducted to investigate the influences of wheat bran fiber (WBF) on growth performance and intestinal epithelium functions in Xiangcun pigs. Twenty Xiangcun pigs with 60 days of age were divided to two groups and exposed to a basal diet (BD) or BD containing 4.3% wheat bran fiber (WFD). WFD improved the average daily gain (ADG) and feed-to-gain ratio (F:G) (p

Variable-flux permanent magnet synchronous machines (PMSMs) which use low-coercive-force permanent magnets have gained a lot of attention as a perspective alternative in the variable-speed driving application for their advantages of operating with improved efficiency in higher speed through adjusting the variable-flux PMSMs’ magnetization states. A hybrid-magnet variable-flux machine which has the variable series-parallel magnetic circuit is investigated. Magnetization state and magnetic circuit type of this variable-flux PMSM are adjustable. This machine’s equivalent magnetic circuit (EMC) model at different magnetization states is established and evaluated. This model can precisely represent the complicated and changeable magnetic circuit in variable-flux PMSMs and provide an effective way for the operating principle and performance analysis. The electromagnetic performances are investigated in detail. The magnetization state variation’s range is wide. Magnetizing/demagnetizing current is small. By fully using reluctance torque, variable-flux PMSM’s torque ability improves greatly. The ratio of high efficiency area in total torque-speed operating area increases obviously. The efficiency during high speed operation also increases. This machine has advantages and overcomes limitations of variable-flux machine which has series magnetic circuit or parallel magnetic circuit.

Colitis is a common and complex intestinal inflammatory disease in which lactate, a metabolite of anaerobic glycolysis, plays a crucial role. Our study aimed to investigate the alleviated effect of lactate in colitis, and to provide a nutritional measure to alleviate colitis injury. The variations in colonic lactate in piglets with DSS-induced colitis were investigated in Experiment 1 (Exp.1). Thirty weaned pigs were allotted into three groups and sampled at different stages of DSS-induced colitis (days 0, 5, and 7). The colonic level of lactate and interleukin 10 (IL-10) was significantly decreased on day 5 when compared to day 0. Colonic lactate, IL-10, and G protein receptor 81 (GPR81) levels were significantly increased on day 7 when compared to day 5. Sixty weaned piglets were assigned to control (basal diet), DSS (basal diet with DSS gavage), or lactate (2% lactate supplementation diet with DSS gavage) groups to investigate the effects of lactate on DSS-induced colitis in Experiment 2 (Exp.2). Lactate reduced the disease activity index (DAI), DSS-induced impairment of colonic structure in response to the critical inflammatory cytokines interleukin 1β (IL-1β) and interleukin 18 (IL-18) when compared with the DSS group. Furthermore, GPR-81 levels, colonic M2 macrophages, and IL-10 levels, the colonic antioxidant capacity, colonic butyrate levels were increased, and eventually improved growth performance post-colitis. The results of this study show that lactate was decreased at the peak of colitis, accumulated in subsidized colitis. Furthermore, dietary lactate supplementation helped to alleviate DSS-induced colitis injury.

Microorganisms drive many important soil processes and ecosystem functioning; however, the soil microbial community can respond to environmental change, which can alter decomposition and storage of soil organic carbon (SOC). We assessed soil microbial taxonomic and functional attributes linked to SOC decomposition and concentration based on a grassland experiment fertilized with nitrogen (N) and phosphorus (P). There were divergent responses at both the taxonomic and functional level to fertilization, with a greater quantitative response in terms of the taxonomic attributes of bacteria and fungi than the abundance of SOC degradation genes. There was a negative correlation between SOC concentration and the abundance of SOC degradation genes. This indicated that N and P fertilization had the potential to stimulate SOC decomposition through modification of the soil microbial community, particularly by increasing the abundance of SOC degradation genes. Redundancy and variation partitioning analyses, as well as structural equation models revealed that soil total P was a pivotal factor shaping the microbial community both for taxonomic and functional gene attributes in the alpine meadow. The abundance of SOC degrading genes had stronger effects on SOC concentrations than did taxonomic attributes. Quantification of the abundance of SOC degradation genes can provide more direct and effective understanding of SOC decomposition processes and prediction of shifts in soil function as organic C pool in grasslands than a focus on the taxonomy of microbial communities. Given the important role of microbial community in regulating SOC decomposition and lower SOC concentrations under N and P fertilization, we recommend that the alpine meadow should be protected from mineral fertilizers, particularly P alone fertilization, to sustain high grassland ecosystem function as SOC sink in this region.

Summary: This protocol describes procedures for quantifying Aspergillus niger growth in both solid and liquid culture. Firstly, by comparing radial growth between mutant and progenitor isolates on solid agar supplemented with sublethal stressors, susceptibility coefficients can be calculated. Secondly, analysis of macromorphological growth types in liquid culture allows full quantification of how a gene of interest affects submerged growth. By combining these assays, an extensive and quantitative dataset of how a gene of interest impacts growth in this fungus is possible.For complete details on the use and execution of this protocol, please refer to Cairns et al. (2019)1 and Cairns et al. (2022).2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Summary: This protocol presents an efficient genetic strategy to investigate gene function in the fungus Aspergillus niger. We combined 5S rRNA-CRISPR-Cas9 technology with Tet-on gene switch to generate conditional-expression mutants via precisely replacing native promoter with inducible promoter. We describe the design and DNA preparation for sgRNAs and donor DNA. We then detail the steps for DNA co-transformation into A. niger protoplasts by PEG-mediated transformation, followed by homozygote isolation. Finally, we describe the genome verification and strain validation of the isolates.For complete details on the use and execution of this protocol, please refer to Zheng et al. (2019).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Pathogenic mitochondrial DNA (mtDNA) mutations can cause a variety of human diseases. The recent development of genome-editing technologies to manipulate mtDNA, such as mitochondria-targeted DNA nucleases and base editors, offer a promising way for curing mitochondrial diseases caused by mtDNA mutations. The CRISPR-Cas9 system is a widely used tool for genome editing; however, its application in mtDNA editing is still under debate. In this study, we developed a mito-Cas9 system by adding the mitochondria-targeted sequences and 3′ untranslated region of nuclear-encoded mitochondrial genes upstream and downstream of the Cas9 gene, respectively. We confirmed that the mito-Cas9 system was transported into mitochondria and enabled knockin of exogenous single-stranded DNA oligonucleotides (ssODNs) into mtDNA based on proteinase and DNase protection assays. Successful knockin of exogenous ssODNs into mtDNA was further validated using polymerase chain reaction-free third-generation sequencing technology. We also demonstrated that RS-1, an agonist of RAD51, significantly increased knockin efficiency of the mito-Cas9 system. Collectively, we provide direct evidence that mtDNA can be edited using the CRISPR-Cas9 system. The mito-Cas9 system could be optimized as a promising approach for the treatment of mitochondrial diseases caused by pathogenic mtDNA mutations, especially those with homoplasmic mtDNA mutations.

Background & Objective: The application of ultrasound imaging for spine evaluation could minimize radiation exposure for patients with adolescence idiopathic scoliosis (AIS). A customized three-dimensional (3D) ultrasound imaging system has been demonstrated to provide reliable and valid coronal curvature measurements. However, these measurements were using the spinous processes as anatomical reference, leading to a predictable underestimation of the traditionally used Cobb angles. An alternative 3D ultrasound image reconstruction method was applied to create coronal images with more lateral features for angle measurement. The objective of this study was to test the reliability and the validity of this angle, the ultrasound curve angle (UCA), and compare the UCA with the Cobb angles on X-ray images of patients with AIS. Materials and methods: This study was divided into: 1) Investigation of intra- and inter-reliability between two raters for measuring the UCA and two operators for acquiring ultrasound images; 2) Investigation of the validity between the radiographic Cobb angle and the UCA. Fifty patients and 164 patients with AIS, were included in the two stages, respectively. Patients underwent bi-planar X-ray and 3D ultrasound scanning on the same day. The proposed UCA was used to measure the coronal curvature from the ultrasound coronal images, which were formed using a newly customized volume projection imaging (VPI) method. The intra-rater/operator and inter-rater and operator reliability of the UCA were tested by intra-class correlation coefficient (ICC) (3,1) and (2,1), respectively. The validity of UCA measurements as compared to radiographic Cobb angles was tested by inter-method ICC (2,1), mean absolute difference (MAD), standard error of measurement (SEM), Pearson correlation coefficient and Bland–Altman statistics. The level of significance was set as 0.05. Results: Excellent intra-rater and intra-operator (ICC (3,1)≥0.973) and excellent inter-rater and inter-operator reliability (ICC (2,1)≥0.925) for UCA measurement, with overall MAD and SEM no more than 3.5° and 1.7° were demonstrated for both main thoracic and (thoraco)lumbar curvatures. Very good correlations were observed between UCA and Cobb angle for main thoracic (R2=0.893) and (thoraco)lumbar (R2=0.884) curves. The mean (SD) measurements in terms of radiographic Cobb and UCA were 27.2 ​± ​11.6° and 26.3 ​± ​11.4° for main thoracic curves; and 26.2 ​± ​11.4° and 24.8 ​± ​9.7° for (thoraco)lumbar curve respectively. One hundred sixty-four subjects (33 male and 131 female subjects; 11–18 years of age, mean of 15.1 ​± ​1.9 years) were included for the validity session. Excellent inter-method variations (ICC (2,K) ≥0.933) with overall MAD and SEM no more than 3.0° and 1.5° were demonstrated for both main thoracic and (thoraco)lumbar curvatures. In addition, Bland–Altman plots demonstrated an acceptable agreement between ultrasound and radiographic Cobb measurements. Conclusion: In this study, very good correlations and agreement were demonstrated between the ultrasound and X-ray measurements of the scoliotic curvature. Judging from the promising results of this study, patients with AIS with different severity of curves can be evaluated and monitored by ultrasound imaging, reducing the usage of radiation during follow-ups. This method could also be used for scoliosis screening.The Translational potential of this article: Ultrasound curve angle (UCA) obtained from 3D ultrasound imaging system can provide reliable and valid evaluation on coronal curvature for patients with AIS, without the need of radiation.

Indoor temperature is a critical environmental factor for piglets that affects the health, welfare, and production efficiency of domesticated pigs. However, there are limited reports about wireless intelligent thermal control and management system for piglets in large-scale pig farms, and evaluation energy saving. This paper presents an Automatic Thermal Control and Management System (ATCMS) of micro-environment for piglets, that could provide a suitable production environment and reduce the energy consumption. ATCMS includes automatic temperature control system (ATCS), multipoint temperature management system (MTMS) and remote access. The infrared heating lamps are chosen in the ATCS for their good performance in heating rate and heating distribution. In order to meet a scientific temperature requirements of piglet growth, the suitable thermal ranges of each growth-stage (STREG) are employed as intelligent control strategies to generate an automatic and precise heating control. MTMS is responsible for communicating with ATCSs and transmitting information to database server for record based on wireless technology. Remote access offers information services for remote administrators by smart phones and clients via Internet. The experiments of ATCMS were carried out both in winter and summer at Binsheng Breeding Piggery Farm, Acheng City, Heilongjiang Province, China. Results showed that the temperature provided by ATCS had been kept in STREG of piglets during the whole winter period. When the indoor temperature during summer period were in or above STREG, heating lamps were turned off by ATCS for a long time. The energy consumption of ATCS were 63.60% in winter and 39.35% in summer with continuous heating system operation. Therefore, ATCMS could take the suitable thermal ranges of each growth-stage as an intelligent control principle to give an automatic and precise control to heating devices in order to meet a scientific temperature requirements of piglet growth.

SCN8A is linked to early infantile epileptic encephalopathy type 13 (EIEE13). We generate a human induced pluripotent stem cell (iPSC) line from a child diagnosed with EIEE, caused by SCN8A variation. The iPSC line expresses high pluripotency markers, retains SCN8A variation and is able to differentiate into three germ layers in vitro. The iPSC lines will provide useful resources for studying the pathogenesis and drug screening of SCN8A-related epilepsy.

Soil salinity and alkalinity can inhibit crop growth and reduce yield, and this has become a global environmental concern. Combined changes in nitrogen (N) application and hill density can improve rice yields in sodic saline–alkaline paddy fields and protect the environment. We investigated the interactive effects of N application rate and hill density on rice yield and N accumulation, translocation and utilization in two field experiments during 2018 and 2019 in sodic saline–alkaline paddy fields. Five N application rates (0 (control), 90, 120, 150, and 180 kg N ha–1 (N0–N4), respectively) and three hill densities (achieved by altering the distance between hills, in rows spaced 30 cm apart: 16.5 cm (D1), 13.3 cm (D2) and 10 cm (D3)) were utilized in a split-plot design with three replicates. Nitrogen application rate and hill density significantly affected grain yield. The mathematical model of quadratic saturated D-optimal design showed that with an N application rate in the range of 0–180 kg N ha–1, the highest yield was obtained at 142.61 kg N ha–1 which matched with a planting density of 33.3×104 ha–1. Higher grain yield was mainly attributed to the increase in panicles m–2. Nitrogen application rate and hill density significantly affected N accumulation in the aboveground parts of rice plants and showed a highly significant positive correlation with grain yield at maturity. From full heading to maturity, the average N loss rate of the aboveground parts of rice plants in N4 was 70.21% higher than that of N3. This is one of the reasons why the yield of N4 treatment is lower than that of the N3 treatment. Nitrogen accumulation rates in the aboveground parts under treatment N3 (150 kg N ha–1) were 81.68 and 106.07% higher in 2018 and 2019, respectively, than those in the control. The N translocation and N translocation contribution rates increased with the increase in the N application rate and hill density, whereas N productivity of dry matter and grain first increased and then decreased with the increase in N application rate and hill density. Agronomic N-use efficiency decreased with an increase in N application rate, whereas hill density did not significantly affect it. Nitrogen productivity of dry matter and grain, and agronomic N-use efficiency, were negatively correlated with grain yield. Thus, rice yield in sodic saline–alkaline paddy fields can be improved by combined changes in the N application rate and hill density to promote aboveground N accumulation. Our study provides novel evidence regarding optimal N application rates and hill densities for sodic saline–alkaline rice paddies.

Abstract A new type of polyhalite potassium ore (NTPPO) was found in the Lower Triassic Jialingjiang Formation, NE Sichuan Basin, SW China. It is water soluble, therefore can be exploited using the water-solution method, and is of great potential of economic value and research significance. Based on cores, thin sections, energy spectrum and SEM analyses, its microfeatures, macrofeatures and origin are discussed, and a genetic model is established to provide a scientific basis for future evaluation, prediction and exploration of potassium ore in the Sichuan Basin. It is proposed that the NTPPO was caused by storm activities: (1) the storm broke the original sedimentary polyhalite–gypsum beds, whose fragments were transported into the salt basin with high content of K+ and Mg2+; (2) in the basin, the polyhalite continued to be formed from gypsum by metasomatism with K- and Mg-rich brine; (3) during diagenesis, under high temperature and high pressure, K–Mg-rich brine from halite continued to replace anhydrite (or gypsum) to form polyhalite.

Hybrid rice significantly contributes to the food supply worldwide. Backbone parents play important roles in elite hybrid rice breeding systems. In this study, we performed pedigree-based analysis of the elite backbone parent rice variety, namely, Shuhui 527 (SH527, Oryza sativa), to exploit key genome regions during breeding. Twenty-four cultivars (including SH527, its six progenitors and 17 derived cultivars) were collected and analyzed with high-density single nucleotide polymorphism (SNP) array. Scanning all these cultivars with genome-wide SNP data indicated the unique contributions of progenitors to the SH527 genome and identified the key genomic regions of SH527 conserved within all its derivatives. These findings were further supported by known rice yield-related genes or unknown QTLs identified by genome-wide association study. This study reveals several key regions for SH527 and provides insights into hybrid rice breeding.

Long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) play an important role in central nervous diseases; however, the exact expression and co-expressed profiles in human traumatic brain injury (TBI) are still unknown. Therefore, we investigated whole blood in 12 patients with TBI and 4 healthy controls to observe expression characteristics with different severity. We identified 3,035 lncRNAs and 1,204 mRNAs differentially expressed in the severe TBI group, 2,362 lncRNAs and 656 mRNAs in the moderate group, and 433 lncRNAs and 100 mRNAs in the mild group. Enrichment analyses showed 30 signaling pathways such as inflammatory and immune response pathways. Subsequently, a lncRNA-gene co-expression network was generated for 717 lncRNA-mRNA pairs and most of them with a positive correlation. Based on GSEA analysis, we found that TBI caused severe immune abnormality reflected on Th1, Th2, and Th17 cell differentiation deficiency. Finally, the expression of one upregulated and one downregulated lncRNA was validated in all three TBI groups, which was consistent with the microarray results. In summary, our results show that expression profiles of lncRNAs and mRNAs are significantly different in bloods from different severity TBI especially in immune response, providing novel insight for lncRNAs in human TBI.

Objective: To investigate the effects of mirror neuron theory-based visual feedback therapy (VFT) on restoration of upper limb function of stroke patients and motor-related cortical function using functional magnetic resonance imaging (fMRI). Methods: Hemiplegic stroke patients were randomly divided into two groups: a VFT group and a control (CTL) group. Sixteen patients in the VFT group received conventional rehabilitation (CR) and VFT for 8 weeks, while 15 patients in the CTL group received only CR. The Barthel Index (BI) was used to assess the activities of daily living at baseline and the 8th week of the recovery training period. The Fugl–Meyer assessment (FMA) scale, somatosensory evoked potential (SEP), and fMRI were used to evaluate the recovery effect of the training therapies. The latencies and amplitudes of N9 and N20 were measured. Before recovery training, fMRI was performed for all patients in the VFT and CTL groups. In addition, 17 patients (9 in the VFT group and 8 in the CTL group) underwent fMRI for follow-up 2 months after treatment. Qualitative data were analyzed using the χ2 test. The independent sample t-test was used to compare normally distributed data among different groups, the paired sample t-test was used to compare data between groups, and the non-parametric test was used to comparing data without normal distribution among groups. Results: There were no significant differences between the VFT and CTL group in all indexes. However, after 8 weeks of recovery training, these indexes were all significantly improved (P

The application of mainstream anammox process is hampered by its overlong start-up and instability under disturbance. A lab-scale mainstream anammox moving bed biofilm reactor (MBBR) was successfully started in 120 days with stepwise decrement in influent nitrogen concentration from sidestream to mainstream condition. The initial colonization by Candidatus Jettenia and filamentous fermenter Anaerolineaceae were potentially mediated by hydrophobic interaction and type IV pilus. Ca. Kuenenia with higher substrate affinity outcompeted Ca. Jettenia, and the predominant fermenters shifted to fermentative Ignavibacteriaceae in the mature biofilm. A novel mainstream anammox biofilm development (MABD) model was constructed to describe biofilm growth, population dynamics, and nitrogen removal performance. The simulation results suggested that higher inocula biomass (460–690 mgVSS·L−1), relative abundance of low-affinity AnAOB in the inocula (e.g., Ca. Jettenia, 1.3–2%), and the early-stage solids retention time (45–68 days) were desired to form thicker biofilm and improve effluent quality during 120-day mainstream anammox MBBR start-up. The mechanistic insights into biofilm formation and predictive power of the newly developed MABD model are of importance to the design and operation of mainstream anammox processes towards more biofilm biomass and higher nitrogen removal efficiency.

Oceanic subduction is an important trigger for mantle heterogeneity, which further increases melt production and controls the compositions of intraplate basalts. Such a role played by the (Paleo-) Pacific subduction have been extensively studied and well constrained based on the widespread mantle xenoliths and intraplate magmatism in the eastern North China Craton. By contrast, the recycled materials from other Phanerozoic subducted slabs beneath the craton are relatively poorly recognized. Here, a reappraisal is made to the recently reported peridotite xenoliths and ~89 Ma host basalts from the Langshan area in the northwest North China Craton and regional data on xenoliths and basalts, with the emphasis on the mass transfer in the mantle wedge from the subducted Paleo-Asian oceanic slab. The Langshan peridotites are fertile in composition and record complex melt extraction and metasomatism. One episode of metasomatism is likely induced by silicate melts with concomitant enrichments in large ion lithophile elements and high field strength elements and positive Eu anomaly, suggestive of the contribution from recycled materials. This metasomatism should take place in Paleozoic according to the diffusion modelling. The host basalts are comparable with partial melts of pyroxenite under 2–3 GPa and have oceanic island basalts-like trace-element compositions. Positive Sr and Eu anomalies, low Rb/Sr and high Ba/Rb ratios, and the moderated depleted but slightly decoupled Sr–Nd isotopes suggest the involvement of subducted oceanic crustal materials in the mantle source. The Pb isotopic compositions are best modeled by the mixing between depleted mantle, altered oceanic crust with minor young (

With the increasing demands for the miniaturization of ferroelectricity-enabled electronic devices, it is highly desirable that the ferroelectricity would exist in low-dimensional materials. However, those ferroelectrics, especially one-dimensional (1D) ones, are still limited. Herein, based on the first-principles calculations, 1D ferroelectricity in novel InSeI nanowires with noncentrosymmetric P42 space group is explored, which could possess three different paths for the reversal of electric polarization. The minimum energy barrier of 97 meV/f.u. for the polarization reversal, comparable with those of other low-dimensional ferroelectrics, could be achieved through a unique four-step concerted process. According to ab initio molecular dynamics simulations, the Curie temperature of the proposed P42-InSeI nanowires is estimated to about 866 K, above room temperature. The electronic structure and carrier mobility of the nanowires are also studied, demonstrating that they could be promising materials in nanoelectronics such as high-density nonvolatile memories due to their stable array structures formed by van der Waals interactions.

Spinal deformities assessment using 3D ultrasound scanning has limitations in fitting onto different back surface contour as well as fitting within the gaps between subject and their spinal brace during bracing assessments. The study proposed a flexible array ultrasound transducer to overcome these limitations.The results demonstrated the feasibility of spinal deformity assessments with a flexible ultrasound array when arranged in four shapes, namely Linear, Concave, Convex, and S-shaped. For comparisons of imaging performance on spinous process using the four shapes, Convex and S-shaped transducer showed a depth dependence and lateral location dependence of the lateral intensity distribution of spinous process, respectively. S-shaped transducer had the least accurate prediction of the location of spinous process, with measurement error of 4.8 ​± ​3.2 ​mm, it also showed poorer prediction on spinal curvature measurements. This is suggested to be due to the asymmetrical distortion to the spinous process due to the lateral location dependence of the image. However, the coronal curve prediction of spine phantom performed well with R-squared values of >0.97 in all transducer shapes.The results of this study paved the way for further investigation on the improvement of image quality and measurement accuracy under different shapes for the flexible array, mechanism of dynamic shape change during the scanning to fit different body contour, as well as extension from 1D to 2D flexible array.

ABSTRACT: Essential oils (EO) are concentrated hydrophobic liquids containing volatile aromatic compounds obtained from plants, which have properties as withdrawn antibiotic growth promoters. The objective of this study was to explore the effects of EO on growth performance, digestibility, immunity and intestinal health in broilers. A total of 500 1-day-old Arbor Acre broilers were randomly put into five groups with 10 replicate cages containing 10 birds each. Birds in the 5 groups were fed a basal diet (CON), and basal diet with 50, 100, 200 or 400 mg/kg EO (EO0.5, EO1, EO2 and EO4) for 42 d respectively. Birds were euthanized at 21d and 42 d, blood and tissue samples were collected. In the study, the digestibility of DM, GE and EE in groups with EO supplementation were significantly increased compared with CON group (P < 0.05). However, only EO2 and EO4 significantly increased the digestibility of CP compared with CON group (P < 0.05). In contrast to CON group, EO0.5 and EO1 in jejunum at 21 d, and EO1 in jejunum at 42 d markedly increased the activity of sucrase (P < 0.05). In addition, the level of SOD of EO2 and EO4 in serum at 21 d was significantly increased compared with CON group (P < 0.05). What's more, the concentration of intestinal mucosa SIgA in jejunum and ileum at 21 d of groups with EO supplementation was significantly increased compared with CON group (P < 0.05). Moreover, V/C in jejunum at 21 d of groups with EO supplementation, CD in jejunum at 42 d was also significantly increased to compare with CON group (P < 0.05). Furthermore, the expression levels of critical genes associated with nutrient transportation (i.e., GLUT2, SGLT1, SLC38A, SLC79A and SLC27A4) and barrier function (TJP1) were quadratically and linearly up-regulated in jejunum and ileum with EO supplementation (P < 0.05). These results suggest that EO has a positive impact on growth, immunity and intestinal health in broilers, and 200 mg/kg of EO was recommended in broiler diet.

To explored the protective effect of manno-oligosaccharides (MOS) on intestinal epithelium exposure to Enterotoxigenic Escherichia coli (ETEC), thirty-two pigs were assigned into four treatments and fed with basal or MOS-containing (0.3% MOS) diet. Pigs were challenged by ETEC or culture medium. Results show that MOS not only reduced diarrhea incidence and fecal E. coli abundance but decreased serum D-lactate and DAO concentrations upon ETEC-challenge (P

In order to effectively evaluate the diet nutritional value of dairy cows, it is essential to accurately predict the diet nutrients digestibility (DND). Conventional predicting DND methods are usually based on the least squares linear regression analysis (LS-LRA), which often relies on a large amount of training samples to accomplish reliable predictions. However, in real-world applications, it is often extremely difficult, costly and time-consuming to obtain a large number of measured samples, especially for the DND prediction of dairy cows. This paper applies a Gaussian process regression (GPR) technique to predict the DND indicators of dairy cows in small samples. To evaluate prediction accuracy effectively, we compared the GPR technique with the LS-LRA, radial basis function artificial neural network (RBF-ANN), support vector regression (SVR) and least squares support vector regression (LS-SVR) methods, using the required sample data obtained from actual digestion experiments. The prediction results indicate that the GPR technique is superior to other conventional methods (especially the LS-LRA method) in predicting the main DND indicators of dairy cows such as dry matter digestibility (DMD), organic matter digestibility (OMD), neutral detergent fiber (NDFD), acid detergent fiber (ADFD), and crude protein digestibility (CPD). It is worth mentioning that the developed GPR-based prediction technique is more suitable for the prediction problems with small samples, which is often the case in the prediction of DND indicators of dairy cows, and then more coincide with actual needs. Keywords: Precision feeding, Dairy cow diet evaluation, Diet nutrients digestibility, Small sample prediction, GPR algorithm

Background: Schizophrenia is a complex mental disorder resulting in poor life quality and high social and economic burden. Despite the fact that genome-wide association studies (GWASs) have successfully identified a number of risk loci for schizophrenia, identifying the causal genes at the risk loci and elucidating their roles in disease pathogenesis remain major challenges. Methods: The summary data-based Mendelian randomization analysis (SMR) was used to integrate a large-scale GWAS of schizophrenia with brain expression quantitative trait loci (eQTL) data and brain methylation expression quantitative trait loci (meQTL) data, to identify novel risk gene(s) for schizophrenia. We then analyzed the mRNA expression and methylation statuses of the gene hit BTN3A2 during the early brain development. Electrophysiological analyses of CA1 pyramidal neurons were performed to evaluate the excitatory and inhibitory synaptic activity after overexpression of BTN3A2 in rat hippocampal slices. Cell surface binding assay was used to test the interaction of BTN3A2 and neurexins. Findings: We identified BTN3A2 as a potential risk gene for schizophrenia. The mRNA expression and methylation data showed that BTN3A2 expression in human brain is highest post-natally. Further electrophysiological analyses of rat hippocampal slices showed that BTN3A2 overexpression specifically suppressed the excitatory synaptic activity onto CA1 pyramidal neurons, most likely through its interaction with the presynaptic adhesion molecule neurexins. Interpretation: Increased expression of BTN3A2 might confer risk for schizophrenia by altering excitatory synaptic function. Our result constitutes a paradigm for distilling risk gene using an integrative analysis and functional characterization in the post-GWAS era. Fund: This study was supported by the Strategic Priority Research Program (B) of the Chinese Academy of Sciences (XDB02020003 to Y-GY), the National Natural Science Foundation of China (31730037 to Y-GY), and the Bureau of Frontier Sciences and Education, Chinese Academy of Sciences (QYZDJ-SSW-SMC005 to Y-GY). Keywords: Schizophrenia, Integrative analysis, BTN3A2, Brain development, Glutamatergic synapse

Spermatogenic dysfunction is a common complication in men with diabetes and is the most important manifestation of diabetes-related male reproduction damage. Astragalin (AG) is one of the main flavonoids from Cuscuta chinensis, which has rich pharmacological activities. This study aimed to establish whether AG may contribute to the recovery from spermatogenic dysfunction. AG (3.3, 10 and 30 mg/kg) and Clomiphene (5 mg/kg) were orally administered to streptozotocin-induced diabetic male mice for 8 weeks. After the experiments performed, reproductive organs, sperm parameters and histomorphological changes were analysed. Antioxidant and anti-inflammatory capacity were estimated in testicular tissues. The results revealed that AG significantly improved the reproductive organs, sperm parameters and testicular morphology to different degrees in diabetic mice. Nitric oxide (NO) and malondialdehyde (MDA) levels were significantly reduced, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), markedly increased in the testicular tissue after AG was administered. Interestingly, AG also downregulated the protein expressions of tumour necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) in testes. In conclusion, AG is a potential beneficial agent to protect diabetic-induced spermatogenic dysfunction in male mice by increasing antioxidant enzymes activities and inhibiting inflammation.

Background: Tracking the spread of the Neisseria gonorrhoeae strains with decreased susceptibility or resistance to cephalosporins is a major priority for global surveillance programmes. Whole-genome sequencing (WGS) has been widely used by increasing countries in North America, Europe, and Pacific to determine the decreased susceptible or resistance determinants of Neisseria gonorrhoeae, track the spread of these determinants throughout the gonococcal population at national or regional level. However, no studies to date have examined the genomic epidemiology of gonorrhea in Asia where the antimicrobial resistant strains of Neisseria gonorrhoeae appears to have emerged before disseminating the strains globally. Methods: We obtained clinical isolates and data from the China Gonococcal Resistance Surveillance Programme (China-GRSP) from 2012 to 2013. We sequenced the genomes of 435 clinical isolates of Neisseria gonorrhoeae, including 112 (25.6%) isolates with decreased susceptibility to ceftriaxone (Cfx-DS). We assessed the association between antimicrobial resistance genotype and phenotype. We also compared our data with the whole genome data of the isolates from the USA and the UK in the GenBank. Findings: The most prevalent MLST STs in our gonococcal population were MLST ST7827 (n = 74), followed by ST7365 (n = 58), ST1600 (n = 38), ST7367 (n = 35), and ST7363 (n = 29). MLST ST1901 which was reported as the predominant ST in the US was not found in our population. A total of 2512 strains, including additional 2077 published NG strains, were further included for phylogenetic analysis. It generated two distinct lineages - lineage 1 and lineage 2. Analysis of MLST ST1901 in the database indicate that most of MLST ST1901 isolates in the lineage2.6 were Cfx-DS isolates while all isolates in the lineage 2.1 were sensitive to ceftriaxone (77/110 vs. 0/13; p

This study investigated molecular mechanisms of lycopene regulating muscle fiber type transformation in vitro and in vivo. Remarkably, TNNI1 protein level was increased, while TNNI2 protein level was decreased in C2C12 myotubes and in mice after lycopene treatment. Lycopene also enhanced enzyme activities of succinic dehydrogenase and malate dehydrogenase and reduced lactate dehydrogenase activity. Meanwhile, lycopene markedly upregulated the protein expressions of P-Akt and P-FoxO1 and decreased FoxO1 protein level. However, specific blocking Akt/FoxO1 signaling by wortmannin effectively eliminated lycopene-induced upregulation of slow MyHC and downregulation of fast MyHC in C2C12 myotubes. In addition, lycopene effectively inhibited miR-22-3p expression in C2C12 myotubes and in mice, whereas miR-22-3p mimics attenuated transformation from fast glycolytic fiber to slow oxidative fiber caused by lycopene. Our findings provided the first evidence that lycopene promoted muscle fiber type transformation from fast-twitch to slow-twitch through Akt/FoxO1 pathway and miR-22-3p.

Summary: Previous work has shown that the paraventricular nucleus of the thalamus (PVT) is an important region that is involved in the conditioned context-induced retrieval of morphine withdrawal memory. However, the upstream neural circuits that activate the PVT to participate in the conditioned context-induced retrieval of morphine withdrawal memory remain unknown. In the present work, we find that the conditioned context activates projection neurons from the prelimbic cortex (PrL) to the PVT, and the inhibition of PrL-PVT projection neurons inhibits the conditioned context-induced retrieval of morphine withdrawal memory; the conditioned context induces an increase in Arc expression, intrinsic excitability, and glutamate output in PrL-PVT projection neurons in morphine-withdrawn mice. These results suggest that the activity of PrL-PVT projection neurons is necessary for the retrieval of morphine withdrawal memory, and the conditioned context causes a plastic change in the activity in these projection neurons during the withdrawal memory retrieval.