Myeloid-Mas Signaling Modulates Pathogenic Crosstalk among MYC+CD63+ Endothelial Cells, MMP12+ Macrophages, and Monocytes in Acetaminophen-Induced Liver Injury.

Acetaminophen overdose is a leading cause of acute liver failure (ALF). Despite the pivotal role of the inflammatory microenvironment in the progression of advanced acetaminophen-induced liver injury (AILI), a comprehensive understanding of the underlying cellular interactions and molecular mechanisms remains elusive. Mas is a G protein-coupled receptor highly expressed by myeloid cells; however, its role in the AILI microenvironment remains to be elucidated. A multidimensional approach, including single-cell RNA sequencing, spatial transcriptomics, and hour-long intravital imaging, is employed to characterize the microenvironment in Mas1 deficient mice at the systemic and cell-specific levels. The characteristic landscape of mouse AILI models involves reciprocal cellular communication among MYC+CD63+ endothelial cells, MMP12+ macrophages, and monocytes, which is maintained by enhanced glycolysis and the NF-κB/TNF-α signaling pathway due to myeloid-Mas deficiency. Importantly, the pathogenic microenvironment is delineated in samples obtained from patients with ALF, demonstrating its clinical relevance. In summary, these findings greatly enhance the understanding of the microenvironment in advanced AILI and offer potential avenues for patient stratification and identification of novel therapeutic targets.

Development and comparative analysis of initiation ability in large-scale Heuchera propagation using tissue culture versus cuttings.

The Heuchera genus, a member of the Saxifragaceae family, encompasses a wide array of varieties and hybrids, serving both traditional medicinal and ornamental purposes. However, a significant knowledge gap persists in achieving efficient mass propagation of diverse Heuchera cultivars creating a substantial market void. To address this, our study focuses on expedited seedling regeneration by investigating leaf cutting and tissue culture techniques to offer novel insights to cultivators. Herein, we successfully rooted thirteen distinct cultivars from the Heuchera and Heucherella (Heuchera × Tiarella) genera through cutting. Moreover, in vitro culture experiments led to the successful induction of calli and shoots from petiole samples. Notably, variations in measured parameters were observed across cultivars in both cutting and tissue culture methodologies. When petiole explants were exposed to cytokinin 6-benzylaminopurine (BA) at concentrations of 0.5, 1.0, and 2.0 mg/L along with auxin α-naphthaleneacetic acid (NAA) at 0.5 mg/L, shoots were produced either directly or indirectly during the primary culture. Exposure to darkness and the application of 2,4-dichlorophenoxyacetic acid (2,4-D) did not promote shoot formation but were beneficial for callus stimulation. Interestingly, a negative correlation was observed between the ease of initiating cutting recovery and inducting tissue culture regeneration, suggesting that cultivars that easily regenerate through cutting might encounter difficulties during induction by tissue culture. In light of these findings, we devised a streamlined and effective protocol for rapid Heuchera propagation. This protocol involves micropropagation, directly acquiring adventitious shoots from primary cultures supplemented by cutting-based propagation methods.

An immunogenic cell death-related classification predicts prognosis and response to immunotherapy in kidney renal clear cell carcinoma.

Introduction: Immunogenic cell death (ICD) is a form of regulated cell death that activates an adaptive immune response in an immunocompetent host and is particularly sensitive to antigens from tumor cells. Kidney clear cell carcinoma (KIRC) is an immunogenic tumor with extensive tumor heterogeneity. However, no reliable predictive biomarkers have been identified to reflect the immune microenvironment and therapeutic response of KIRC. Methods: Therefore, we used the CIBERSORT and ESTIMATE algorithms to define three ICD clusters based on the expression of ICD-related genes in 661 KIRC patients. Subsequently, we identified three different ICD gene clusters based on the overlap of differentially expressed genes (DEGs) within the ICD clusters. In addition, principal component analysis (PCA) was performed to calculate the ICD scores. Results: The results showed that patients with reduced ICD scores had a poorer prognosis and reduced transcript levels of immune checkpoint genes regulated with T cell differentiation. Furthermore, the ICD score was negatively correlated with the tumor mutation burden (TMB) value of KICD. patients with higher ICD scores showed clinical benefits and advantages of immunotherapy, indicating that the ICD score is an accurate and valid predictor to assess the effect of immunotherapy. Discussion: Overall, our study presents a comprehensive KICD immune-related ICD landscape that can provide guidance for current immunotherapy and predict patient prognosis to help physicians make judgments about the patient's disease and treatment modalities, and can guide current research on immunotherapy strategies for KICD.

A case-control evaluation of Spasm control and Tolerability of the Modified Atkins diet versus classic ketogenic diet in Chinese Children with infantile epileptic spasms syndrome.

OBJECTIVE: This study was conducted to compare the efficacy, tolerability and safety of the modified Atkins diet (MAD) and the classic ketogenic diet (KD) in Chinese children with infantile epileptic spasms syndrome. METHODS: We retrospectively recruited and analyzed 56 children with infantile epileptic spasms syndrome started on the MAD (n = 21) and classic KD (n = 35) at our institution from June 2016 to July 2022. RESULTS: The MAD group had exhibited comparable rates of spasm reduction (>50%) as the classic KD group at the time point of 3 months (66.7% for MAD, 75% for KD; p = 0.510), 6 months (75% for MAD, 82.6% for KD; p = 0.694), and 12 months (84.6% for MAD, 90.9% for KD; p = 1.000). The probability of patients remaining on the MAD was higher (p = 0.048) than those consuming the classic KD. By analyzing discontinuation reasons, we found that the MAD patients had a lower rate of poor compliance than the classic KD patients (p = 0.014). Response to the diet at 1 month and fewer anti-seizure medicines (ASMs) had tried before KD initiation were possible factors in regard to likelihood of spasm-free response to the diet therapy at 3 months (p = 0.001 and p = 0.014, respectively). CONCLUSIONS: The efficacy of spasm control was similar in the MAD, with better tolerability and higher compliance compared to the classic KD. Therefore, MAD could be the primary treatment for children in China with- infantile epileptic spasms syndrome. Additionally, an earlier beginning of the diet treatment may have significant advantages.

CsMYB15 positively regulates Cs4CL2-mediated lignin biosynthesis during juice sac granulation in navel orange.

'Lane Late', a late-maturing navel orange cultivar, is mainly distributed in the Three Gorges Reservoir area, which matures in the late March of the next year and needs overwintering cultivation. Citrus fruit granulation is a physiological disorder, which is characterized by lignification and dehydration of juice sac cells, seriously affecting the commercial value of citrus fruits. The pre-harvest granulation of late-maturing navel orange is main caused by low temperature in the winter, but its mechanism and regulation pattern remain unclear. In this study, a SG2-type R2R3-MYB transcription factor, CsMYB15, was identified from Citrus sinensis, which was significantly induced by both juice sac granulation and low temperature treatment. Subcellular localization analysis and transcriptional activation assay revealed that CsMYB15 protein was localized to the nucleus, and it exhibited transcriptional activation activity in yeast. Over-expression of CsMYB15 by stable transformation in navel orange calli and transient transformation in kumquat fruits and navel orange juice sacs significantly increased lignin content in the transgenic lines. Further, Yeast one hybrid, EMSA, and LUC assays demonstrated that CsMYB15 directly bound to the Cs4CL2 promoter and activated its expression, thereby causing a high accumulation of lignin in citrus. Taken together, these results elucidated the biological function of CsMYB15 in regulating Cs4CL2-mediated lignin biosynthesis, and provided novel insight into the transcriptional regulation mechanism underlying the juice sac granulation of late-maturing navel orange.

Study on wetting mechanism of nonionic silicone surfactant on coal dust.

Coal dust disasters are serious in coal mining. The use of nonionic surfactants can effectively improve the wettability of coal dust and reduce the content of suspended coal dust in the air. For the problem of low wettability of ordinary surfactants, this paper selects silicone surfactants with high surface activity and low surface tension to improve the wetting ability of coal dust. To explore the wettability of nonionic silicone surfactants on coal dust, the effects of six nonionic silicone surfactants on the wettability of coal dust surfaces were studied by experiments. The test objects were four kinds of coal samples with different metamorphic degrees. The surface tension, wetting time, and contact angle experiments were carried out, and the critical micelle concentration and the expansion coefficient of the coal surface were calculated. The wetting time of the compound solution was measured to verify the synergistic effect of the compound solution. The results show that: 6 # has the best wetting effect on coal dust, followed by 4 # and 2 #; The order of surface tension is: 1 # < 3 # < 4 # < 6 # < 5 # < 2 #, the surface tension of 1 # is the lowest (19.962 mN/m); 1 # and 4 # are easier to spread on the surface of coal dust, the spreading coefficient of coking coal is the largest and the contact angle is the smallest, which is 18.8°. The 4 # and 6 # with a mass ratio of 8:2 were compounded. The compound surfactant solution had a significant synergistic effect. Compared with the monomer surfactant solution, the wettability of long-flame coal and coking coal increased by 15.14% and 10.00%, respectively. The results of this study can provide reference and experimental support for the development of high-efficiency dust suppressants based on silicone surfactants.

ATP6V0C gene variants were identified in individuals with epilepsy, with or without developmental delay.

The cause of epilepsy with or without developmental disorders was unidentified in a significant proportion of patients. Whole exome sequencing was performed in three unrelated patients with early-onset epilepsy, with or without developmental delay and intellectual disability. We identified de novo heterozygous variants (p.Arg119Trp, p.Val99_Ser102del, c.260_263 + 11delinsGCCCA) in the ATP6V0C gene, which encodes a subunit of vacuolar ATPase. Three-dimensional protein modeling showed that the variant p.Arg119Trp in ATP6V0C affected the hydrogen bonds with the 115th and 123rd residues, and the protein stability. The p.Val99_Ser102del and c.260_263 + 11delinsGCCCA variants in the other two patients resulted in a loss of function with microdeletion or splicing effects. Their seizures and psychomotor developmental outcomes were different, and all patients had a good prognosis. Our study provides evidence that de novo heterozygous ATP6V0C variants are related to epilepsy and associated with or without developmental delay.

Mass spectrometry imaging and single-cell transcriptional profiling reveal the tissue-specific regulation of bioactive ingredient biosynthesis in Taxus leaves.

Taxus leaves provide the raw industrial materials for taxol, a natural antineoplastic drug widely used in the treatment of various cancers. However, the precise distribution, biosynthesis, and transcriptional regulation of taxoids and other active components in Taxus leaves remain unknown. Matrix-assisted laser desorption/ionization-mass spectrometry imaging analysis was used to visualize various secondary metabolites in leaf sections of Taxus mairei, confirming the tissue-specific accumulation of different active metabolites. Single-cell sequencing was used to produce expression profiles of 8846 cells, with a median of 2352 genes per cell. Based on a series of cluster-specific markers, cells were grouped into 15 clusters, suggesting a high degree of cell heterogeneity in T. mairei leaves. Our data were used to create the first Taxus leaf metabolic single-cell atlas and to reveal spatial and temporal expression patterns of several secondary metabolic pathways. According to the cell-type annotation, most taxol biosynthesis genes are expressed mainly in leaf mesophyll cells; phenolic acid and flavonoid biosynthesis genes are highly expressed in leaf epidermal cells (including the stomatal complex and guard cells); and terpenoid and steroid biosynthesis genes are expressed specifically in leaf mesophyll cells. A number of novel and cell-specific transcription factors involved in secondary metabolite biosynthesis were identified, including MYB17, WRKY12, WRKY31, ERF13, GT_2, and bHLH46. Our research establishes the transcriptional landscape of major cell types in T. mairei leaves at a single-cell resolution and provides valuable resources for studying the basic principles of cell-type-specific regulation of secondary metabolism.

Genome-wide identification and expression analysis of TCP family genes in Catharanthus roseus.

Introduction: The anti-tumor vindoline and catharanthine alkaloids are naturally existed in Catharanthus roseus (C. roseus), an ornamental plant in many tropical countries. Plant-specific TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) transcription factors play important roles in various plant developmental processes. However, the roles of C. roseus TCPs (CrTCPs) in terpenoid indole alkaloid (TIA) biosynthesis are largely unknown. Methods: Here, a total of 15 CrTCP genes were identified in the newly updated C. roseus genome and were grouped into three major classes (P-type, C-type and CYC/TB1). Results: Gene structure and protein motif analyses showed that CrTCPs have diverse intron-exon patterns and protein motif distributions. A number of stress responsive cis-elements were identified in promoter regions of CrTCPs. Expression analysis showed that three CrTCP genes (CrTCP2, CrTCP4, and CrTCP7) were expressed specifically in leaves and four CrTCP genes (CrTCP13, CrTCP8, CrTCP6, and CrTCP10) were expressed specifically in flowers. HPLC analysis showed that the contents of three classic TIAs, vindoline, catharanthine and ajmalicine, were significantly increased by ultraviolet-B (UV-B) and methyl jasmonate (MeJA) in leaves. By analyzing the expression patterns under UV-B radiation and MeJA application with qRT-PCR, a number of CrTCP and TIA biosynthesis-related genes were identified to be responsive to UV-B and MeJA treatments. Interestingly, two TCP binding elements (GGNCCCAC and GTGGNCCC) were identified in several TIA biosynthesis-related genes, suggesting that they were potential target genes of CrTCPs. Discussion: These results suggest that CrTCPs are involved in the regulation of the biosynthesis of TIAs, and provide a basis for further functional identification of CrTCPs.

Integrated mass spectrometry imaging and single-cell transcriptome atlas strategies provide novel insights into taxoid biosynthesis and transport in Taxus mairei stems.

Taxol, which is a widely used important chemotherapeutic agent, was originally isolated from Taxus stem barks. However, little is known about the precise distribution of taxoids and the transcriptional regulation of taxoid biosynthesis across Taxus stems. Here, we used MALDI-IMS analysis to visualize the taxoid distribution across Taxus mairei stems and single-cell RNA sequencing to generate expression profiles. A single-cell T. mairei stem atlas was created, providing a spatial distribution pattern of Taxus stem cells. Cells were reordered using a main developmental pseudotime trajectory which provided temporal distribution patterns in Taxus stem cells. Most known taxol biosynthesis-related genes were primarily expressed in epidermal, endodermal, and xylem parenchyma cells, which caused an uneven taxoid distribution across T. mairei stems. We developed a single-cell strategy to screen novel transcription factors (TFs) involved in taxol biosynthesis regulation. Several TF genes, such as endodermal cell-specific MYB47 and xylem parenchyma cell-specific NAC2 and bHLH68, were implicated as potential regulators of taxol biosynthesis. Furthermore, an ATP-binding cassette family transporter gene, ABCG2, was proposed as a potential taxoid transporter candidate. In summary, we generated a single-cell Taxus stem metabolic atlas and identified molecular mechanisms underpinning the cell-specific transcriptional regulation of the taxol biosynthesis pathway.

Pinellia genus: A systematic review of active ingredients, pharmacological effects and action mechanism, toxicological evaluation, and multi-omics application.

The dried tuber of Pinellia ternata (Thunb.) Breit, Pinelliae Rhizoma (PR, also named 'Banxia' in Chinese), is widely used in traditional medicine. This review aims to provide detail summary of active ingredients, pharmacological effects, toxic ingredients, detoxification strategies, and omic researches, etc. Pharmacological ingredients from PR are mainly classified into six categories: alkaloids, amino acids, polysaccharides, phenylpropanoids, essential oils, and glucocerebrosides. Diversity of chemical composition determines the broad-spectrum efficacy and gives a foundation for the comprehensive utilization of P. ternata germplasm resources. The pharmacological compounds are involved in inhibition of cancer cells by targeting various pathways, including activation of immune system, inhibition of proliferation and cycle, induction of apoptosis, and inhibition of angiogenesis. The pharmacological components of PR act on nervous system by targeting neurotransmitters, activating immune system, decreasing apoptosis, and increasing redox system. Lectins, one major class of the toxic ingredients extracted from raw PR, possess significant toxic effects on human cells. Inflammatory factors, cytochrome P450 proteins (CYP) family enzymes, mammalian target of rapamycin (mTOR) signaling factors, transforming growth factor-ß (TGF-ß) signaling factors, and nervous system, are considered to be the target sites of lectins. Recently, omic analysis is widely applied in Pinellia genus studies. Plastome genome-based molecular markers are deeply used for identifying and resolving phylogeny of Pinellia genus plants. Various omic works revealed and functional identified a series of environmental stress responsive factors and active component biosynthesis-related genes. Our review summarizes the recent progress in active and toxic ingredient evaluation, pharmacological effects, detoxification strategies, and functional gene identification and accelerates efficient utilization of this traditional herb.

First report of Neopestalotiopsis rosae causing leaf blight on Shatangju in southern China.

Shatangju (Citrus reticulata Blanco cv. Shatangju) belongs to genus Citrus and was cultivated extensively in southern China. In April 2022, a leaf blight-like symptom (firstly brown spots appeared on infecting leaves, then these brown spots extended, finally the whole leaves displayed blight-like symptom) was observed on 5%~10% of Shatangju seedlings (around five hundreds in total) in an orchard located in Wuhan city, Hubei, China. Diseased leaves from three seedlings were collected and cut into pieces (0.2 to 0.5 cm). These pieces were surface-sterilized using 75% ethanol for 3 min, rinsed with sterile distilled water for several times, then placed on potato dextrose agar (PDA) and incubated at 26°C with 12-h light/dark cycle. Over 20 pieces plated, wherein 30% were identified as Colletotrichum fructicola, 60% as Neopestalotiopsis spp., and 10% developed saprophytes. C. fructicola was a known pathogen on citrus, thus Neopestalotiopsis spp. was further investigated. Eight single-conidium colonies of the Neopestalotiopsis spp. were obtained, wherein STJ-8 was chosen as a representative for further study. The average growth rate of STJ-8 was 15.1±0.5 mm/day (n=5). Fungal colonies produced white cottony mycelium with abundant black acervuli distributed in concentric rings 6-8 days after planting, which ranged from 342.3 to 710.5 µm in diameter (n=100). Conidia were fusoid, five cells, four septa with average dimensions of 25.36×5.47 µm (n=100). Basal and apical cells were hyaline, wherein three middle cells were brown with darker septa. The apical cell was cylindrical with two to three transparent accessory filaments (13.7 to 30.5 µm in length, n=80). Basal cell was conic with an appendage (4.1 to 8.8 µm in length, n=40). Partial sequences of internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF-1α), and ß-tubulin (TUB2) were amplified with reported primers (White et al. 1990; Lee et al. 2006; Maharachchikumbura et al. 2014), sequenced, and submitted to GenBank (accession nos. ITS: OP236541; TEF-1α: OP250124; TUB2:OP263094). BLASTn results showed 100% identity with the corresponding sequences of Neopestalotiopsis rosae. A multilocus phylogenetic analysis showed STJ-8 was closest to N. rosae. Thus, STJ-8 was identified as N. rosae. Pathogenicity tests were performed on one-year-old Shatangju seedlings and detached primary leaves by inoculating needle-wounded leaves with seven days old 5-mm mycelial plugs/acervuli (about 5000 spores) of STJ-8. Control seedlings/leaves were inoculated with 5-mm PDA plugs/sterile water drops. All inoculated detached leaves were cultured at same the place with STJ-8 cultured, while inoculated seedlings were put in a growth chamber at 26°C under a 16-h light/dark cycle (60% humidity). Symptoms developed on all inoculated leaves (except healthy control) 2 and 4 days post-inoculation by mycelial plugs and acervuli, respectively. N. rosae was re-isolated from the inoculated leaves, confirming Koch's postulates. N. rosae has been reported to cause diseases on various plants worldwide (Rebollar-Alviter et al. 2020; Xavier et al. 2021; Lawrence et al. 2022). In China, N. rosae has been reported to cause leaf spot/blight on pecan and strawberry (Wu et al. 2021; Gao et al. 2022), which caused great loss on these crops. To our knowledge, this is the first report of N. rosae causing leaf disease on citrus. Our study is important for developing control strategies against N. rosae in future.

Diagnosis of Neurological Infections in Pediatric Patients from Cell-Free DNA Specimens by Using Metagenomic Next-Generation Sequencing.

Central nervous system (CNS) infections can cause significant morbidity and mortality, especially in children. Rapid and accurate pathogenic detection in suspected CNS infections is essential for disease control at the early stage of infection. To evaluate the performance of metagenomic next-generation sequencing (mNGS) of cell-free DNA (cfDNA) in cerebrospinal fluid (CSF) in pediatric patients, we retrospectively analyzed the efficiency of cfDNA mNGS in children with CNS infections (n = 257) or noninfectious neurological disorders (n = 81). The CSF samples of 124 random subjects were used to evaluate the accuracy between mNGS of cfDNA and whole-cell DNA (wcDNA). In total, cfDNA mNGS detected a wide range of microbes with a detection rate of 71.0%, and the sensitivity and total coincidence rate with clinical diagnosis reached 68.9% and 67.5%, respectively. Compared with wcDNA mNGS, cfDNA mNGS had a higher efficacy in detecting viruses (66 versus 13) and Mycobacterium (7 versus 1), with significantly higher reads per million. The dominant causative pathogens were bacteria and viruses in CNS infections, but these presented with different pathogen spectra in different age categories. The best timing for the mNGS test ranged from 1 to 6 days after the start of anti-infection therapy, and the earlier mNGS started, the better was identification of bacterial CNS infections. This study emphasized that cfDNA mNGS had overall superiority to conventional methods on causative pathogen detection in pediatric CNS infections, and it was even better than wcDNA mNGS. Furthermore, research needs to be better validated in large-scale clinical trials to improve the clinical applications of cfDNA mNGS. IMPORTANCE Our study emphasized that cfDNA mNGS had overall superiority to conventional methods on causative pathogen detection in CNS-infected children, and it was even better than wcDNA mNGS. cfDNA mNGS detected a wide range of pathogens with a high total coincidence rate (67.5%) against clinical diagnosis. The best timing for cfDNA mNGS detection ranged from 1 to 6 days, rather than 0 days, after the start of empirical anti-infection therapy. The earlier mNGS started, the better the identifications of bacterial CNS infections. To the best of our knowledge, this research is the first report evaluating the clinical utility of mNGS with different methods (cfDNA versus wcDNA) of extracting DNA from CSF specimens in diagnosing pediatric CNS infections. Meanwhile, this is the largest cohort study that has evaluated the performance of mNGS using cfDNA from CSF specimens in pediatric patients with CNS infections.

Identification and functional analysis of calcium sensor calmodulins from heavy metal hyperaccumulator Noccaea caerulescens.

Noccaea caerulescens (J. Presl & C. Presl) F. K. Mey. is a heavy metal hyperaccumulator exhibiting extreme tolerance to various environmental stresses. To date, the functional role of Ca2+ -binding protein in this plant is largely unknown. To investigate the function of calmodulins (CaMs) in N. caerulescens , CaM2 , a Ca2+ sensor encoding gene, was identified and functionally characterised. Protein structure analysis showed that NcCaM2 contains four classic exchange factor (EF)-hand motifs with high sequence similarity to the CaM proteins from model plant Arabidopsis thaliana L. Tissue specific expression analysis showed that NcCaM2 is constitutively expressed in stems, leaves, and roots. Expression level of NcCaM2 was significantly upregulated under various environmental stimulus, indicating a potential involvement of NcCaM2 in the tolerance to abiotic stresses. The heterologous expression of NcCaM2 in a yeast mutant strain increased the heavy metal tolerance in yeast cells. Furthermore, the constitutive expression of NcCaM2 enhanced the heavy metal tolerance capability of transgenic tobacco (Nicotiana tabacum L.) plants. Our data suggested an important role of NcCaM2 in the responses to environmental stresses and provided a potential target gene to enhance of the ability to hyperaccumulate metals.

Inhibition of secretory leukocyte protease inhibitor (SLPI) promotes the PUMA-mediated apoptosis and chemosensitivity to cisplatin in colorectal cancer cells.

BACKGROUND: Aberrant expression of Secretory Leukocyte Protease Inhibitor (SLPI) has been associated with human cancer growth and its suppression was identified as a potential target for anti-cancer drugs, particularly in colorectal cancer. However, the underlying mechanism by which SLPI affected the development of drug resistance in CRC remains unclear. OBJECTIVE: This study investigated the role of SLPI in the p53-up-regulated modulator of apoptosis (PUMA)-mediated CRC cells' apoptosis and their chemosensitivity to Cisplatin. METHODS: A series of qRT-PCR and western blot analyses were performed to characterize the expressions of SLPI, PUMA, and Akt in CRC lines. Tunel, transwell, and CCK-8 analyses were monitored to define the impacts of the siRNA-mediated knockdown of SLPI on CRC cell development. Furthermore, in vivo development of CRC was evaluated in nude mice infected with siSLPI or Cisplatin alone or both, and Ki67 and caspase-3 immunohistochemistry assay was monitored on multiple tissue microarray from the same cohort. RESULTS: Our results showed that SLPI inhibition strongly promoted the expressions of the pro-apoptotic protein PUMA, cleaved-caspase3 and Bax and reduced the cell viability of HT29 and HT116 cell lines in vitro. In addition, siSLPI knockdown effectively suppressed both Akt and FoxO3 proteins and improved the sensitivity to cisplatin chemotherapy. Xenograft tumor assay revealed a lowered growth in mice treated with Cisplatin, while combined treatment of siSLPI achieved more significant anticancer effects than Cisplatin alone. CONCLUSIONS: Taken together, these findings demonstrated that suppression of SLPI might repress the growth of human colorectal cancer cells both in vitro and in vivo. These results suggested SLPI as a novel resistance factor to Cisplatin, and a combination of Cisplatin and SLPI inhibitor be beneficial for colorectal cancer therapy.

Hepatocyte-specific Mas activation enhances lipophagy and fatty acid oxidation to protect against acetaminophen-induced hepatotoxicity in mice.

BACKGROUND & AIMS: Acetaminophen (APAP) is the most common cause of drug-induced liver injury (DILI); however, treatment options are limited. Mas is a G protein-coupled receptor whose role in APAP-induced hepatotoxicity has not yet been examined. METHODS: Intrahepatic Mas expression was determined in both human and mouse DILI models. Mas1-/-, AlbcreMas1f/f, Ppara-/-, Mas1-/-Ppara-/- and wild-type mice were challenged with APAP for the in vivo analyses of Mas-AKT-FOXO1 axis-dependent lipophagy and fatty acid oxidation (FAO), using pharmacological compounds and genetic tools. Liver samples were collected for RNA-sequencing, proteomics, metabolomics, lipidomics, and metabolic flux analysis. Live-imaging of liver and histological, biochemical, and molecular studies were performed to evaluate APAP-induced hepatotoxicity in mice. Primary hepatocytes and hepatic cell lines were exposed to APAP for in vitro analysis. RESULTS: Intrahepatic Mas expression was significantly upregulated in human and mouse DILI models. Mice with systemic, liver-specific, or hepatocyte-specific Mas1 deficiency were vulnerable to APAP-induced hepatotoxicity. They exhibited substantially impaired lipophagy and downstream FAO, which was accompanied by the activation of AKT and suppression of FOXO1. In addition, the prophylactic activation of Mas conferred strong protection against APAP challenge in mice, with remarkably enhanced lipophagy and FAO dependent on the AKT-FOXO1 axis. Moreover, the protective effects of AVE0991 were substantially diminished by the inhibition of either lipophagy or FAO. CONCLUSIONS: The activation of Mas on hepatocytes enhanced AKT-FOXO1-dependent lipophagy and downstream FAO, protecting mice from APAP-induced hepatotoxicity and indicating that hepatocyte-specific Mas might be a novel therapeutic target for DILI. IMPACT AND IMPLICATIONS: Mas signalling arises as a novel therapeutic target for patients with APAP-induced liver injury. The Mas-AKT/FOXO1-fatty acid degradation pathway could be critical for the development of treatment strategies for APAP overdose. When Mas signalling is targeted, the extent of liver injury should be considered at the time of administration. These findings obtained from APAP-challenged mice still need to be confirmed in a clinical context.

The effects of fucoidan as a dairy substitute on diarrhea rate and intestinal barrier function of the large intestine in weaned lambs.

This paper explores the effects of fucoidan on the frequency of diarrhea, colon morphology, colon antioxidant status, cytokine content, short-chain fatty acids, and microflora of cecal contents in early weaned lambs in order to provide a reference for the intestinal health of young ruminants. Fucoidan is a natural active polysaccharide extracted from kelp and other large brown algae. It has many biological effects, such as improving immunity, nourishing the stomach and intestines, and anti-tumor properties. This study investigated the effects of fucoidan supplementation in milk replacer on the large intestine's ability to act as an intestinal barrier in weaned lambs. With six duplicate pens and one lamb per pen, a total of 24 weaned lambs (average starting body weight of 7.32 ± 0.37 kg) were randomly assigned to one of four milk replacer treatments. Four concentrations of fucoidan supplementation (0, 0.1, 0.3, and 0.6% dry matter intake) were employed to investigate the effects of fucoidan on cecal fermentation and colon microbial organization. The test period lasted 37 days (1 week before the test and 1 month after the test), and lamb cecal contents and colon organization were collected for examination. In addition, the fecal status of all lambs was observed and recorded daily, allowing us to calculate the incidence of diarrhea in weaned lambs. The findings demonstrated that fucoidan may significantly increase the concentration of short-chain fatty acids (propionic acid and butyric acid) in the cecal digesta of weaned lambs. In weaned lambs, 16S rDNA testing showed that fucoidan at 0.3-0.6% (dry matter intake) was beneficial for boosting the variety of the intestinal bacteria and modifying the relative abundance of a few bacterial strains. In addition, fucoidan enhanced colon antioxidant and immune functions and decreased the diarrhea rate to relieve weaning stress. This result demonstrates that milk replacer supplementation with fucoidan contributes to the improvement in the large intestinal health of weaned lambs.

Emerging starch composite nanofibrous films for food packaging: Facile construction, hydrophobic property, and antibacterial activity enhancement.

Polymers synthesized from green resources have many advantages in food packaging and hence their development is very important. Herein, starch/polyvinyl alcohol (PVA) nanofibrous composite films were fabricated by electrospinning technology. Steam-induced cross-linking reaction with glutaraldehyde (GTA) and silver sodium zirconium phosphate (Ag-ZrP) was employed to improve the hydrophobic and antibacterial properties of the constructed nanofibrous films, respectively. The effects of starch/PVA ratio on the micro-morphology and mechanical properties of the binary composite film were investigated. The composite film showed optimal uniformity, bead-free electrospun nanofibers, with enhanced mechanical strength for the 60/40 (v/v) starch/PVA composite. Moreover, the crystallinity of PVA was reduced during the electrospinning process, whereas the introduction of PVA strengthened the hydrogen interactions and improved the thermal stability of the composite films. After the cross-linking with GTA, the starch/PVA films became more hydrophobic. Furthermore, the starch/PVA films embedded with Ag-ZrP had outstanding antibacterial property against both Gram-negative and Gram-positive bacteria. This work demonstrated the potential prospects of electrospun starch nanofibrous films in the food packaging field.

First report of Fusarium oxysporum and Fusarium solani causing root rot on trifoliate orange rootstock in China.

Trifoliate orange (Poncirus trifoliata L) is a thorny tree of the Rue family, which is extensively used as citrus rootstock in China. In January 2021, several leaf yellowing, declining, and wilting citrus seedlings grafted on trifoliate orange rootstock with rotted main roots were observed in orchards located in Wuhan city, Hubei, China. In old orchards, the incidence of diseased roots was approximately 90%. Diseased roots from seven plants were collected and cut into small pieces (0.2 to 0.5 cm). These pieces were then surface-sterilized using 0.1% mercury bichloride for 3 min, 75% ethanol for 3 min, rinsed with sterile distilled water for several times, and then placed on potato dextrose agar (PDA) supplemented with 0.05% lactic acid (v/v), and incubated at at 25±2°C in dark. Fifty-threesingle-conidium isolates with morphological characteristics similar to Fusarium spp. were obtained (Leslie and Summerell 2006), which displayed two kinds of colony morphology. Thirty isolates showed white to orange-white abundant aerial mycelium in rings and acquired a yellow to orange pigmentation, tweenty-three isolates showed white to pink, fluffy aerial mycelium in rings and acquired an orange to red pigmentation. Isolate WG-1 and HrmY-9 from each group were used for future identification. The average colony growth rate of WG-1 and HrmY-9 on PDA was 0.95±0.06 and 0.69±0.11 mm/day, n=4, respectively. WG-1 produced numerous oval, unicellular microconidia without septa, 4.03-9.87×1.01-5.13 µm, n=80 and very few macroconidia with two to four septa, narrowed at both ends, 11.08-22.64×1.67-4.91 µm, n=30. HrmY-9 produced numerous curved macroconidia with three to four septa, 18.03-37.33×2.16-7.8 µm, n=80, microconidia were unicellular, oval, and 5.33-16.19×1.74-6.51 µm, n=50. Sequences of internal transcribed spacer (ITS), translation elongation factor 1-alpha (EF-1α), and DNA-directed RNA polymerase largest subunit (RPB1) genes were amplified with the primers ITS1/ITS4, EF1a-F/EF1a-R, and RPB1-F5/RPB1-R8, respectively (White et al. 1990, O'Donnell et al. 1998, O'Donnell et al. 2010), sequenced and deposited in GenBank. Sequences of isolate WG-1 (GenBank accession No. ON045437, ON063232 and ON089664) and HrmY-9 (GenBank accession No. ON045438, ON063233 and ON089665) were 100% identical with the corresponding sequences of Fusarium oxysporum (OM876904, JF430180, and MT568959) and F. solani (MT605584, MK617767, and MT305110), respectively. Based on above results, WG-1 and HrmY-9 was identified as F. oxysporum and F. solani, respectively. Pathogenicity test were performed on healthy one-year-old trifoliate orange seedlings by dipping their injured roots into conidial suspension (50 ml, 1×106 conidia/mL) for 1 h and the rest of conidial suspension was added to the pot after replanting to make sure the inoculum was in contact with the roots. Roots of control plants were inoculated with sterilized water. All experiments were repeated twice. All plants were cultured at 26°C under a 16-h light/dark cycle. Typical symptoms developed on most of inoculated seedlings two months post inoculation. No disease symptoms appeared on control plants. Same colonies were reisolated from the inoculated roots, confirming Koch's postulates. To our knowledge, this is the first report of F. oxysporum and F. solani causing root rot on trifoliate orange rootstock in China. The identification of F. oxysporum and F. solani as the causal agents of the observed root rot on trifoliate orange rootstock is critical to the prevention and control of this disease in the future.

Comparative study on the pyrolysis behavior and pyrolysate characteristics of Fushun oil shale during anhydrous pyrolysis and sub/supercritical water pyrolysis.

Injected steam can be converted to the sub/supercritical state during the in situ exploitation of oil shale. Thus, the pyrolysis behavior and pyrolysate characteristic of Fushun oil shale during anhydrous pyrolysis and sub/supercritical water pyrolysis were fully compared. The results revealed that the discharged oil yields from sub/supercritical water pyrolysis were 5.44 and 14.33 times that from anhydrous pyrolysis at 360 °C and 450 °C, which was due to the extraction and driving effect of sub/supercritical water. Also, sub/supercritical water could facilitate the discharge and migration of shale oil from the pores and channels. The H2 and CO2 yields in sub/supercritical water pyrolysis were higher than that in anhydrous pyrolysis, resulting from the water-gas shift reaction. The component of shale oil was dominated by saturated hydrocarbons in anhydrous pyrolysis, which accounted for 50-65%. In contrast, a large amount of asphaltenes and resins was formed during pyrolysis in sub/supercritical water due to the solvent effect and weak thermal cracking. The shale oil from anhydrous pyrolysis was lighter than that from sub/supercritical water pyrolysis. Sub/supercritical water reduced the geochemical characteristic indices and lowered the hydrocarbon generation potential and maturity of solid residuals, which can be attributed to the fact that more organic matter was depolymerized and released. The pyrolysate characteristic of oil shale in sub/supercritical water pyrolysis was controlled by multiple mechanisms, including solvent and driving effect, chemical hydrogen-donation and acid-base catalysis.