Morphological, molecular, and functional characterization of mouse glutamatergic myenteric neurons.

The enteric nervous system (ENS) functions largely independently of the central nervous system (CNS). Glutamate, the dominant neurotransmitter in the CNS and sensory afferents, is not a primary neurotransmitter in the ENS. Only a fraction (∼2%) of myenteric neurons in the mouse distal colon and rectum (colorectum) are positive for vesicular glutamate transporter type 2 (VGLUT2), the structure and function of which remain undetermined. Here, we systematically characterized VGLUT2-positive enteric neurons (VGLUT2-ENs) through sparse labeling with adeno-associated virus, single-cell mRNA sequencing (scRNA-seq), and GCaMP6f calcium imaging. Our results reveal that the majority of VGLUT2-ENs (29 of 31, 93.5%) exhibited Dogiel type I morphology with a single aborally projecting axon; most axons (26 of 29, 89.7%) are between 4 and 10 mm long, each traversing 19 to 34 myenteric ganglia. These anatomical features exclude the VGLUT2-ENs from being intrinsic primary afferent or motor neurons. The scRNA-seq conducted on 52 VGLUT2-ENs suggests different expression profiles from conventional descending interneurons. Ex vivo GCaMP6f recordings from flattened colorectum indicate that almost all VGLUT2-EN (181 of 215, 84.2%) are indirectly activated by colorectal stretch via nicotinic cholinergic neural transmission. In conclusion, VGLUT2-ENs are a functionally unique group of enteric neurons with single aborally projecting long axons that traverse multiple myenteric ganglia and are activated indirectly by colorectal mechanical stretch. This knowledge will provide a solid foundation for subsequent studies on the potential interactions of VGLUT2-EN with extrinsic colorectal afferents via glutamatergic neurotransmission.NEW & NOTEWORTHY We reveal that VGLUT2-positive enteric neurons (EN), although constituting a small fraction of total EN, are homogeneously expressed in the myenteric ganglia, with a slight concentration at the intermediate region between the colon and rectum. Through anatomic, molecular, and functional analyses, we demonstrated that VGLUT2-ENs are activated indirectly by noxious circumferential colorectal stretch via nicotinic cholinergic transmission, suggesting their participation in mechanical visceral nociception.

Utilizing serum metabolomics for assessing postoperative efficacy and monitoring recurrence in gastric cancer patients.

OBJECTIVE: (1) This study aims to identify distinct serum metabolites in gastric cancer patients compared to healthy individuals, providing valuable insights into postoperative efficacy evaluation and monitoring of gastric cancer recurrence; (2) Methods: Serum samples were collected from 15 healthy individuals, 16 gastric cancer patients before surgery, 3 months after surgery, 6 months after surgery, and 15 gastric cancer recurrence patients. T-test and analysis of variance (ANOVA) were performed to screen 489 differential metabolites between the preoperative group and the healthy control group. Based on the level of the above metabolites in the recurrence, preoperative, three-month postoperative, and six-month postoperative groups, we further selected 18 significant differential metabolites by ANOVA and partial least squares discriminant analysis (PLS-DA). The result of hierarchical clustering analysis about the above metabolites showed that the samples were regrouped into the tumor-bearing group (comprising the original recurrence and preoperative groups) and the tumor-free group (comprising the original three-month postoperative and six-month postoperative groups). Based on the results of PLS-DA, 7 differential metabolites (VIP > 1.0) were further selected to distinguish the tumor-bearing group and the tumor-free group. Finally, the results of hierarchical clustering analysis showed that these 7 metabolites could well identify gastric cancer recurrence; (3) Results: Lysophosphatidic acids, triglycerides, lysine, and sphingosine-1-phosphate were significantly elevated in the three-month postoperative, six-month postoperative, and healthy control groups, compared to the preoperative and recurrence groups. Conversely, phosphatidylcholine, oxidized ceramide, and phosphatidylglycerol were significantly reduced in the three-month postoperative, six-month postoperative, and healthy control groups compared to the preoperative and recurrence groups. However, these substances did not show significant differences between the preoperative and recurrence groups, nor between the three-month postoperative, six-month postoperative, and healthy control groups; (4) Conclusions: Our findings demonstrate the presence of distinct metabolites in the serum of gastric cancer patients compared to healthy individuals. Lysophosphatidic acid, triglycerides, lysine, sphingosine-1-phosphate, phosphatidylcholine, oxidized ceramide, and phosphatidylglycerol hold potential as biomarkers for evaluating postoperative efficacy and monitoring recurrence in gastric cancer patients. These metabolites exhibit varying concentrations across different sample categories.

The role of microbiota in gastric cancer: A comprehensive review.

BACKGROUND: Gastric cancer (GC) continues to pose a significant global threat in terms of cancer-related fatalities. Despite notable advancements in medical research and therapies, further investigation is warranted to elucidate its underlying etiology and risk factors. Recent times have witnessed an escalated emphasis on comprehending the role of the microbiota in cancer development. METHODS: This review briefly delves into recent developments in microbiome-related research pertaining to gastric cancer. RESULTS: According to studies, the microbiota can influence GC growth by inciting inflammation, disrupting immunological processes, and generating harmful microbial metabolites. Furthermore, there is ongoing research into how the microbiome can impact a patient's response to chemotherapy and immunotherapy. CONCLUSION: The utilization of the microbiome for detecting, preventing, and managing stomach cancer remains an active area of exploration.

A pomegranate seed-structured nanozyme-based colorimetric immunoassay for highly sensitive and specific biosensing of Staphylococcus aureus.

Staphylococcus aureus (S. aureus) infections are a serious threat to human health. The development of rapid and sensitive detection methods for pathogenic bacteria is crucial for accurate drug administration. In this research, by combining the advantages of enzyme-linked immunosorbent assay (ELISA), we synthesized nanozymes with high catalytic performance, namely pomegranate seed-structured bimetallic gold-platinum nanomaterials (Ps-PtAu NPs), which can catalyze a colorless TMB substrate into oxidized TMB (oxTMB) with blue color to achieve colorimetric analysis of S. aureus. Under the optimal conditions, the proposed biosensor could quantitatively detect S. aureus at levels ranging from 1.0 × 101 to 1.0 × 106 CFU mL-1 with a limit of detection (LOD) of 3.9 CFU mL-1. Then, an integrated color picker APP on a smartphone enables on-site point-of-care testing (POCT) of S. aureus with LOD as low as 1 CFU mL-1. Meanwhile, the proposed biosensor is successfully applied to the detection of S. aureus in clinical samples with high sensitivity and specificity.

A predictive nomogram developed and validated for gastric cancer patients with triple-negative tumor markers.

Aim: To predict the prognosis of gastric cancer patients with triple-negative tumor markers. Materials & methods: Prognostic factors of the nomogram were identified through univariate and multivariate Cox regression analyses. Calibration and receiver operating characteristic curves were used to assess accuracy. Decision curve analysis and concordance indexes were utilized to compare the nomogram with the pathological tumor, node, metastasis stage. Results: A nomogram incorporating log odds of positive lymph nodes, tumor size and lymphocyte-to-monocyte ratio was constructed. The calibration and receiver operating characteristic curves (area under the curve >0.85) showed high accuracy in predicting overall survival. The concordance indexes (0.832 vs 0.760; p < 0.001) and decision curve analysis demonstrated that the nomogram was superior to the pathological tumor, node, metastasis stage. Conclusion: A prediction and risk stratification nomogram has been developed and validated for gastric cancer patients with triple-negative tumor markers.

Effects of carbon emission trading system on corporate green total factor productivity: Does environmental regulation play a role of green blessing?

Extant studies focus on the impact of environmental regulation on regional economic growth or environmental pollution, and a lot of research outcomes have been made. However, from the perspective of corporate green sustainable development, the question of whether carbon emission trading represents a "green blessing" remains unclear. To address this issue, we employ a staggered difference-in-differences model to investigate the effects and mechanisms of the carbon emissions trading pilot policy (CETPP) on the green total factor productivity (GTFP) of listed manufacturing companies in China. Our results demonstrate that: a) CETPP can effectively promote corporate GTFP, and the robustness of this result is verified through a series of checks; b) the mediating role of environmental, social, and governance (ESG) performance is critical in the relationship between CETPP and corporate GTFP, with environmental and governance performance serving as two key transmission channels; and c) CEO green experience and public environmental concern both play the moderating roles on the relationship between CETPP and GTFP; d) CETPP has a stronger positive impact on GTFP of private enterprises and enterprises in the maturity life cycle; and e) CETPP has a spatial spillover effect on GTFP, and the effect will decay as spatial distance increases. Our study offers both theoretical and practical implications for enterprises to achieve their green economic development objectives, so as to promote China's high-quality development.

Association of levels of metabolites with the safe margin of rectal cancer surgery: a metabolomics study.

BACKGROUND: Rectal cancer is one of the most lethal of gastrointestinal malignancies. Metabonomics has gradually developed as a convenient, inexpensive and non-destructive technique for the study of cancers. METHODS: A total of 150 tissue samples from 25 rectal cancer patients were analyzed by liquid chromatography-mass spectrometry (LC-MS), and 6 tissue samples were collected from each patient (group 1: tumor; group 2: 0.5 cm from tumor; group 3:1 cm from tumor; group 4:2 cm from tumor; group 5:3 cm from tumor and group 6:5 cm from tumor). The differential metabolites of tumor tissues and 5 cm from the tumor (normal tissues) were first selected. The differential metabolites between tumor tissues and normal tissues were regrouped by hierarchical clustering analysis, and further selected by discriminant analysis according to the regrouping of clustering results. The potential safe margin of clinical T(cT)1,cT2 stage rectal cancer and cT3,cT4 stage rectal cancer at the metabolomic level was further identified by observing the changes in the level of differential metabolites within the samples from group 1 to group 6. RESULTS: We found 22 specific metabolites to distinguish tumor tissue and normal tissue. The most significant changes in metabolite levels were observed at 0.5 cm (cT1, cT2) and 2.0 cm (cT3, cT4) from the tumor, while the changes in the tissues afterwards showed a stable trend. CONCLUSIONS: There are differential metabolites between tumor tissues and normal tissues in rectal cancer. Based on our limited sample size, the safe distal incision margin for rectal cancer surgery in metabolites may be 0.5 cm in patients with cT1 and cT2 stage rectal cancer and 2.0 cm in patients with cT3 and cT4 stage rectal cancer.

Structural characterization and biological activities of a new polysaccharide isolated from Morchella Sextelata.

Morchella is the famous medicinal fungi in the ascomycetes. In this study, a new water-soluble polysaccharide (MSP-3-1) with an average molecular weight of 2.35 × 107 Da was extracted and purified from fruiting bodies of cultivated M. Sextelata. The structural characterization and biological activities of purified polysaccharide was further investigated. The results indicated that MSP-3-1 was mainly a α-glucan, mainly consisting of mannose (Man), glucose (Glc) and galactose (Gal) in a ratio of 5.10: 91.39: 3.51. Its surface morphology exhibited irregular lamellar structures with small voids. And the particle size analysis showed that MSP-3-1 was the homogeneous nanoparticle in water solution. Furthermore, the antioxidant activity analysis showed that MSP-3-1 possessed certain scavenging activity against hydroxyl radicals, DPPH radicals and ABTS radicals in a dose-dependent manner. Immunological tests suggested that MSP-3-1 could significantly promote the proliferation, phagocytosis and nitric oxide (NO) production of macrophage RAW264.7. Thus, our results will provide a theoretical basis for the development and utilization of Morchella Sextelata polysaccharides as an immunmodulatory component in functional foods.

Discovery of 4-phenylindolines containing a (5-cyanopyridin-3-yl)methoxy moiety as potent inhibitors of the PD-1/PD-L1 interaction.

With the great success of anti-programmed cell death-1 (PD-1)/programmed cell death ligand-1 (PD-L1) monoclonal antibodies in clinical applications, blocking the PD-1/PD-L1 pathway has become the most compelling strategy in the field of tumor immunotherapy. In this study, a novel series of 4-phenylindolines containing a (5-cyanopyridin-3-yl)methoxy moiety were developed, and their structure-activity relationships were preliminarily discussed. Among them, compounds M17 and M23 exhibited the most potent ability to disrupt the PD-1/PD-L1 interaction, demonstrating IC50 values of 60.1 nM and 53.2 nM, respectively. The binding mode of M23 was further explored by molecular docking analysis with dimeric PD-L1. Therefore, M17 and M23 are promising lead compounds for developing potent inhibitors of the PD-1/PD-L1 axis.

Characterization of chikusetsusaponin IV and V induced apoptosis in HepG2 cancer cells.

BACKGROUND: Chikusetsusaponin IV and V (CsIV and CsV), two typical oleanolic acid saponins, are mainly derived from the rhizome of Panax japonicus C.A. Mey. To reveal the anti-cancer effect of CsIV and CsV on liver cancer cells, human hepatic cancer cell lines (HepG2) were exposed to these saponins, and various physiological responses of HepG2 were investigated. METHODS AND RESULTS: HepG2 cells were treated with CsIV, CsV and 5-fluorouracil (5-FU). Cell proliferation was measured by CCK-8 assay. The cell cycle arrest, cell apoptosis and intracellular Ca2+ levels were respectively identified by flow cytometry. The mitochondrial membrane potential was detected by fluorescence microscopy. And, the levels of apoptosis-related proteins were analyzed by western blotting. Both CsIV and CsV were demonstrated to inhibit cell viability, and induce cell cycle arrest and apoptosis of HepG2 in a dose-dependent manner. They also enhanced the intracellular Ca2+ level and decreased the mitochondrial membrane potential in HepG2 cells. Furthermore, p53 and p21 were found up-regulated in HepG2 cells treated by CsIV and CsV. The apoptotic proteins, bax, cytochrome c, cleaved caspase-3/-9, were all found activated in HepG2 cells after CsIV and CsV treatment. The anti-apoptotic protein, bcl-2, was significantly down-regulated in all treated HepG2 cells. CONCLUSION: Our data demonstrated that CsIV and CsV exerted significant cytotoxic effects on HepG2 cells without affecting normal liver cells. And, these chikusetsusaponins, especially for CsIV, showed a potent effect on promoting cell apoptosis in HepG2 cells, which was associated with the activation of p53-mediated apoptosis pathway.

Association of perfusate oxygenation with cardiovascular disorder in tetralogy of fallot children: A nested case-control study.

BACKGROUND: Little is known regarding whether hyperoxic reoxygenation was associated with higher risk of cardiovascular disorder following tetralogy of Fallot repair. METHODS: We performed a nested case-control study among patients aged 1 month-18 years undergoing complete repair of tetralogy of Fallot in 2012-2018. We measured the highest perfusate oxygenation (PpO2) during aortic occlusion in 107 cardiovascular disorder cases and in 321 controls matched 1:3 to the cases on date of surgery, sex, and area of residence. We analyzed the association between PpO2 and outcome using multivariable conditional logistic regression adjusted for covariates. We further identified and integrated the risk covariates to build prediction nomograms. RESULTS: Cases had higher percentage of exposure to PpO2 > 200 mmHg (86.0% vs. 76.1%, p = .019) than controls. Patients with PpO2 > 200 mmHg had an increased risk of cardiovascular disorder compared to those with PpO2 ≤ 200 mmHg (odd ratio [OR] = 2.075, 95% confidence interval [CI] = 1.035, 4.158, p = .039) adjusted for matching, clinical and procedural covariates. Categorical PpO2, lower body mass index, lower SpO2, untreated minor aortopulmonary collateral arteries, high immediately postoperative central venous pressure, and longer cardiopulmonary bypass time were independent risk factors for cardiovascular disorder (all p < .05). Combining PpO2 nomogram slightly improved discrimination compared with covariate-based nomogram alone for training cohort (area under receiver operating characteristic curve [AUC] = 0.768 vs. 0.761) and for internal validation (AUC = 0.759 vs. 0.753). CONCLUSION: Our findings suggest association exists between high PpO2 during aortic occlusion and cardiovascular disorder risk, and nomogram integrating clinical and procedural factors may be useful in management of patients with tetralogy of Fallot.

Comparative transcriptome analysis of rhizome nodes and internodes in Panax. japonicus var. major reveals candidate genes involved in the biosynthesis of triterpenoid saponins.

The rhizome of P. japonicus var. major, one of the important herbs in Traditional Chinese medicine (TCM), has been used as tonic and hemostatic drugs in Tujia and Miao ethnic groups of China for thousand years. In this study, comparative metabolite and transcriptome analysis of rhizome nodes and internodes of wild P. japonicus var. major was performed to reveal their different roles in the biosynthesis of triterpene saponins. The results showed that the node was the crucial section for the synthesis of ginsenosides in the rhizome. The content of oleanane-type ginsenosides in the node was much higher than those in the internode. Most isoprenoid biosynthesis-related genes were highly expressed in the node. And, candidate UDP-glycosyltransferase (UGT) genes were also found to be differentially expressed between node and internode. Our study will provide a better understanding of the metabolism of ginsenosides in the rhizome of P. japonicus var. major.

Full-length transcriptome sequencing and modular organization analysis of oleanolic acid- and dammarane-type saponins related gene expression patterns in Panax japonicus.

The saponins found in Panax japonicus, a traditional medicinal herb in Asia, exhibit high degrees of structural and functional similarity. In this study, metabolite analysis revealed that oleanolic acid-type and dammarane-type saponins were distributed unevenly in three tissues (rhizome_Y, rhizome_O, and secRoot) of P. japonicus. Single-molecule real-time (SMRT) sequencing and next generation sequencing (NGS) data revealed distinct and tissue-specific transcriptomic patterns relating to the production of these two types of saponins. In the co-expression network and hierarchical clustering analyses, one 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and two 1-deoxy-D-xylulose-5-phosphate synthase (DXS) etc. transcripts were found to be key genes associated with the biosynthesis of oleanolic acid and dammarane-type saponins in P. japonicus, respectively. In addition, cytochrome p450 (CYP) and UDP-glucuronosyltransferase (UGT) family proteins that serve as regulators of saponin biosynthesis-related genes were also found to exhibit tissue-specific expression patterns. Together these results offer a comprehensive metabolomic and transcriptomic overview of P. japonicus.

High phosphate removal using La(OH)3 loaded chitosan based composites and mechanistic study.

Our present study was to prepare a biomass-supported adsorbents with high adsorptive capacity and high selectivity to prevent the accelerated eutrophication in water body. To this end, different metal hydroxide (La, Zr and Fe) first was successfully loaded on chitosan microspheres. Then the quaternary ammonium group with different content was introduced into the adsorbent by polymerization. By comparison of adsorption properties, chitosan-La(OH)3-quaternary ammonium-20% (CS-La-N-20%) has strong adsorption to phosphate (160 mg/g) by immobilizing nano-sized La(OH)3 within a quaternary-aminated chitosan and it maintain high adsorption in the presence of salt ions. The pH results indicated that the CS-La-N-20% would effectively sequestrate phosphate over a wide pH range between 3 and 7 without significant La3+ leaching. What's more, adsorption capacity on the introduce of positively charged quanternary-aminated groups was significantly higher than that of the unmodified adsorbents at alkaline conditions. The column adsorption capacity reached 1300 bed volumes (BV) when phosphate concentration decreased until 0.5 mg/L at 6 BV/hr. The column adsorption/desorption reveals that no significant capacity loss is observed, indicating excellent stability and repeated use property. Characterizations revealed that phosphate adsorption on CS-La-N-20% through ligand exchange (impregnated nano-La(OH)3) and electrostatic attraction (positively charged quanternary-aminated groups). All the results suggested that CS-La-N-20% can serve as a promising adsorbent for preferable phosphate removal in realistic application.

Mannosylated structures of mycobacterial lipoarabinomannans facilitate the maturation and activation of dendritic cells.

Lipoarabinomannan (LAM) is an important virulent factor secreted by mycobacteria, which generally elicit a strong immune response in the host. In this study, the structural difference of LAMs from three mycobacterial strains, Mycobacterium tuberculosis H37Rv, Mycobacterium smegmatis mc2155 and a newly discovered clinical isolate, M. sp. QGD101, was analyzed and further evaluated whether these LAMs can induce DC maturation and promote the immunomodulatory properties. The results reveal that the major structural difference of these LAMs is the amount of mannosyl residues, especially at the terminal end of LAM, which play a key role in determining the divergent response of DCs after mycobacterial infection. Also, this study indicates an important relevance between the glycosylated structure of LAM and its immunomodulatory property, which is helpful to develop a potential approach for identification of different mycobacteria and also lays a foundation for the development of a novel polysaccharide immunological strategy against tuberculosis.

Human gingiva-derived mesenchymal stem cells alleviate inflammatory bowel disease via IL-10 signalling-dependent modulation of immune cells.

Current evidence indicates that inflammatory bowel disease (IBD) is caused primarily by impaired mucosal immunity, resulting in an imbalance between epithelial barrier function and tissue inflammation. Human gingiva-derived mesenchymal stem cells (GMSCs) exhibit immunomodulatory and anti-inflammatory effects in a variety of immunity- and inflammation-associated diseases. However, the role of GMSCs in treating IBD has not been elucidated. Our study, therefore, examined the therapeutic effect and mechanism of GMSCs in a murine colitis model of IBD. Our results indicate that the infusion of GMSCs significantly prolonged survival and relieved symptoms. Phenotype analyses showed that the frequencies of NK1.1+ and CD11b+ cells, as well as CD4 T cells in the spleen, were suppressed in GMSC-treated mice compared with the PBS- or fibroblast-treated control groups. Additionally, GMSC treatment markedly increased the numbers of interleukin (IL)-10+ regulatory T cells, reduced the secretion of pro-inflammatory cytokines, and increased production of anti-inflammatory cytokines. A mechanistic study revealed that anti-IL-10R antibody abolished the protective effect of GMSCs compared with mice treated with anti-IgG antibody. Thus, our results indicate that GMSCs play a critical role in alleviating colitis by modulating inflammatory immune cells via IL-10 signalling.

Interaction between tissue factor pathway inhibitor-2 gene polymorphisms and environmental factors associated with coronary atherosclerosis in a Chinese Han.

To investigate the association of single nucleotide polymorphisms (SNPs) within tissue factor pathway inhibitor-2 (TFPI-2) gene polymorphisms and additional gene-environment interaction with coronary atherosclerosis risk. Generalized multifactor dimensionality reduction (GMDR) was used to screen the best interaction combination among 4 SNPs, smoking and alcohol drinking. Logistic regression was performed to investigate association between 4 SNPs within TFPI-2 gene and coronary atherosclerosis risk. Coronary atherosclerosis risk was significantly higher in carriers with the A allele of rs34489123 within TFPI-2 gene than those with GG genotype (GA+AA versus GG), adjusted OR (95% CI) = 1.70 (1.20-2.31), and was also higher in carriers with the G allele of rs4264 within TFPI-2 gene than those with AA genotype (AG+GG versus AA), adjusted OR (95% CI) = 1.62 (1.21-2.11). GMDR model shown the best models for gene-environment interaction were rs34489123 and smoking after adjusting the covariates, which scored 10 out of 10 for cross-validation consistency and 0.0010 for the sign test. Heavy LD was found for SNPs rs34489123 and rs59805398 (D' value was more than 0.8). Compared to control individuals, the AG haplotypes appeared to be significantly associated with increased coronary atherosclerosis risk, OR (95% CI) = 1.73 (1.22-2.32). We found that the A allele of rs34489123 and the G allele of rs4264 within TFPI-2 gene, interaction between rs34489123 and smoking and AG haplotypes were all associated with increased coronary atherosclerosis risk.

[Study on interaction between ginsenosides Rg_1,Rb_1 and Ro and bovine serum albumin].

Small molecules with physiological or pharmacological activities need to interact with biological macromolecules in order to function in the body. As the protein with the highest proportion of plasma protein,serum albumin is the main protein binding to various endogenous or exogenous small molecules. Serum albumin interacts with small molecules in a reversible non-covalent manner and transports small molecules to target sites. Bovine serum albumin( BSA) is an ideal target protein for drug research because of its low cost and high homology with human serum albumin. The research on the interaction between drugs and BSA has become a hotspot in the fields of pharmacy,medicine,biology and chemistry. In this research,molecular docking method was used to study the interaction between three small ginsenosides with high pharmacological value( Rg_1,Rb_1,Ro) and bovine serum albumin( BSA),and the binding mode information of three ginsenosides interacting with BSA was obtained. The results of molecular docking showed that ginsenosides and amino acid residues in the active pocket of proteins could be combined by hydrophobic action,hydrogen bonding and electrostatic action. The interaction between small ginsenosides and bovine serum albumin is not the only form,and their interaction has many forms of force. The interaction between these molecules and various weak forces is the key factor for the stability of the complex. The results of this study can provide the structural information of computer simulation for the determination of the interaction patterns between active components and proteins of ginseng.

Lineage specification revealed by single-cell gene expression analysis in porcine preimplantation embryos.

After zygotic genome activation and lineage specification, zygotes develop into late blastocysts comprising three distinct cell types. The molecular mechanisms underlying this progress are largely unknown in pigs. Here, we intended to analyze an extensive set of regulators at the single-cell level to define the events involved in the development of the porcine blastocysts. Using a quantitative microfluidics approach in single cells, we detected mRNA levels of 96 genes known to function in early embryonic development and maintenance of stem cell pluripotency simultaneously in 480 individual cells derived from porcine preimplantation embryos. The developmental transitions can be distinguished based on distinctive gene expression profiles, and we identified paired box 6 (PAX6) and aquaporin 3 (AQP3) expressed in early and late developmental stages, respectively. Two lineages can be segregated in porcine early and late blastocysts by the expression patterns of lineage-specific genes such as DAB2, clathrin adaptor protein (DAB2) for trophectoderm (TE), platelet derived growth factor receptor alpha (PDGFRA), Nanog homeobox (NANOG), fibronectin 1 (FN1), hepatocyte nuclear factor 4 alpha (HNF4A), goosecoid homeobox (GSC), nuclear receptor subfamily 5 group A member 2 (NR5A2), and lysine acetyltransferase 6A (KAT6A; previously known as MYST3) for inner cell mass (ICM). However, the epiblast and primitive endoderm cannot be identified in late blastocysts, and those TE or ICM lineage-specific genes were low expressed in blastomeres from the morula. Our results shed light on early cell fate determination in porcine preimplantation embryos and offer theoretical support for deriving porcine embryonic stem cells.

The Phosphatome of Medicinal and Edible Fungus Wolfiporia cocos.

Wolfiporia cocos is an important medicinal and edible fungus that grows in association with pine trees, and its dried sclerotium has been used as a traditional medicine in China for centuries. However, the commercial production of W. cocos sclerotia is currently limited by shortages in pine wood resources. Since protein phosphatases (PPs) play significant roles in growth, signal transduction, development, metabolism, sexual reproduction, cell cycle, and environmental stress responses in fungi, the phosphatome of W. cocos was analyzed in this study by identifying PP genes, studying transcript profiles and assigning PPs to orthologous groups. Fifty-four putative PP genes were putatively identified in W. cocos genome based on homologous sequences searching using BLASTx program against the Saccharomyces cerevisiae, Fusarium graminearum, and Sclerotinia sclerotiorum databases. Based on known and presumed functions of orthologues of these PP genes found in other fungi, the putative roles of these W. cocos PPs in colonization, hyphal growth, sclerotial formation, secondary metabolism, and stress tolerance to environment were discussed in this study. And the level of transcripts from PP genes in the mycelium and sclerotium stages was also analyzed by qRT-PCR. Our study firstly identified and functional discussed the phosphatome in the medicinal and edible fungus W. cocos. The data from our study contribute to a better understanding of PPs potential roles in various cellar processes of W. cocos, and systematically provide comprehensive and novel insights into W. cocos economically important traits that could be extended to other fungi.