MiR-100-5p-rich small extracellular vesicles from activated neuron to aggravate microglial activation and neuronal activity after stroke.

Ischemic stroke is a common cause of mortality and severe disability in human and currently lacks effective treatment. Neuronal activation and neuroinflammation are the major two causes of neuronal damage. However, little is known about the connection of these two phenomena. This study uses middle cerebral artery occlusion mouse model and chemogenetic techniques to study the underlying mechanisms of neuronal excitotoxicity and severe neuroinflammation after ischemic stroke. Chemogenetic inhibition of neuronal activity in ipsilesional M1 alleviates infarct area and neuroinflammation, and improves motor recovery in ischemia mice. This study identifies that ischemic challenge triggers neuron to produce unique small extracellular vesicles (EVs) to aberrantly activate adjacent neurons which enlarge the neuron damage range. Importantly, these EVs also drive microglia activation to exacerbate neuroinflammation. Mechanistically, EVs from ischemia-evoked neuronal activity induce neuronal apoptosis and innate immune responses by transferring higher miR-100-5p to adjacent neuron and microglia. MiR-100-5p can bind to and activate TLR7 through U18U19G20-motif, thereby activating NF-κB pathway. Furthermore, knock-down of miR-100-5p expression improves poststroke outcomes in mice. Taken together, this study suggests that the combination of inhibiting aberrant neuronal activity and the secretion of specific EVs-miRNAs may serve as novel methods for stroke treatment.

Injectable hydrogel encapsulating siMMP13 with anti-ROS and anti-apoptotic functions for osteoarthritis treatment.

BACKGROUND: Osteoarthritis (OA) is a degenerative joint disease characterized by the progressive degeneration of articular cartilage, leading to pain, stiffness, and loss of joint function. The pathogenesis of OA involves multiple factors, including increased intracellular reactive oxygen species (ROS), enhanced chondrocyte apoptosis, and disturbances in cartilage matrix metabolism. These processes contribute to the breakdown of the extracellular matrix (ECM) and the loss of cartilage integrity, ultimately resulting in joint damage and dysfunction. RNA interference (RNAi) therapy has emerged as a promising approach for the treatment of various diseases, including hATTR and acute hepatic porphyria. By harnessing the natural cellular machinery for gene silencing, RNAi allows for the specific inhibition of target genes involved in disease pathogenesis. In the context of OA, targeting key molecules such as matrix metalloproteinase-13 (MMP13), which plays a critical role in cartilage degradation, holds great therapeutic potential. RESULTS: In this study, we developed an innovative therapeutic approach for OA using a combination of liposome-encapsulated siMMP13 and NG-Monomethyl-L-arginine Acetate (L-NMMA) to form an injectable hydrogel. The hydrogel served as a delivery vehicle for the siMMP13, allowing for sustained release and targeted delivery to the affected joint. Experiments conducted on destabilization of the medial meniscus (DMM) model mice demonstrated the therapeutic efficacy of this composite hydrogel. Treatment with the hydrogel significantly inhibited the degradation of cartilage matrix, as evidenced by histological analysis showing preserved cartilage structure and reduced loss of proteoglycans. Moreover, the hydrogel effectively suppressed intracellular ROS accumulation in chondrocytes, indicating its anti-oxidative properties. Furthermore, it attenuated chondrocyte apoptosis, as demonstrated by decreased levels of apoptotic markers. CONCLUSION: In summary, the injectable hydrogel containing siMMP13, endowed with anti-ROS and anti-apoptotic properties, may represent an effective therapeutic strategy for osteoarthritis in the future.

Integrative leaf anatomy structure, physiology, and metabolome analyses revealed the response to drought stress in sainfoin at the seedling stage.

INTRODUCTION: Sainfoin (Onobrychis viciaefolia) is a vital legume forage, and drought is the primary element impeding sainfoin growth. OBJECTIVE: The anatomical structure, physiological indexes, and metabolites of the leaves of sainfoin seedlings with a drought-resistant line of P1 (DRL) and a drought-sensitive material of 2049 (DSM) were analyzed under drought (-1.0 MPa) with polyethylene glycol-6000 (PEG-6000). METHODS: The leaf anatomy was studied by the paraffin section method. The related physiological indexes were measured by the hydroxylamine oxidation method, titanium sulfate colorimetric method, thiobarbituric acid method, acidic ninhydrin colorimetric method, and Coomassie brilliant blue method. The metabolomics analysis was composed of liquid chromatography tandem high-resolution mass spectrometry (LC-MS/MS). RESULTS: The results revealed that the thickness of the epidermis, palisade tissue, and sponge tissue of DRL were significantly greater than those of DSM. The leaves of DRL exhibited lower levels of superoxide anion (O2 •-) production rate, hydrogen peroxide (H2O2) content, and malondialdehyde (MDA) content compared with DSM, while proline (Pro) content and soluble protein (SP) content were significantly higher than those of DSM. A total of 391 differential metabolites were identified in two samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that the primary differential metabolites were concentrated into the tyrosine metabolism; isoquinoline alkaloid biosynthesis; ubiquinone and other terpenoid quinone biosynthesis; neomycin, kanamycin, and gentamicin biosynthesis; and anthocyanin biosynthesis metabolic pathways. CONCLUSION: Compared with DSM, DRL had more complete anatomical structure, lower active oxygen content, and higher antioxidant level. The results improved our insights into the drought-resistant mechanisms in sainfoin.

Molecular cloning and functional characterization in response to saline-alkali stress of the MhZEP gene in Arabidopsis thaliana.

Soil salinization is one of the major environmental factors that restrict plant growth and development. Zeaxanthin epoxidase (ZEP) functions in ABA biosynthesis and the xanthophyll cycle and has a vital role in plant responses to various environmental stresses. It was found by quantitative real-time PCR (qRT-PCR) that MhZEP responded to saline-alkali stress and showed the highest expression at 48 h of saline-alkali stress, which was 14.53-fold of 0 h. The MhZEP gene was cloned from the apple rootstock begonia (Malus halliana Koehne) and its protein physicochemical properties were analyzed. Subsequently, the functional characterization of MhZEP (ID: 103403091) was further investigated in Arabidopsis thaliana. The MhZEP contained a complete open reading frame with a length of 1998 bp, and encoded 665 amino acids with an isoelectric point of 7.18. Phylogenetic tree analysis showed that MhZEP was the most homologous and closely related to Glycine max. Compared with wild-type, transgenic plants grew better under saline-alkali stress and the MhZEP-OE line showed higher chlorophyll content, carotenoid content, enzyme activities (POD, SOD, CAT and APX) and K+ content, whereas they had lower chlorosis and Na+ content than the wild type (WT), which indicated that they had strong resistance to stress. The expression levels of saline-alkali stress-related genes in A. thaliana MhZEP-OE were examined by qRT-PCR, and it was found that the MhZEP improved the tolerance of A. thaliana to saline-alkali stress tolerance by regulating the expression of carotenoid synthesis genes (MhPSY, MhZDS, MhLYCB and MhVDE) and ABA biosynthesis genes (MhNCED5, MhABI1 and MhCYP707A2). And the potassium-sodium ratio in the cytoplasm was increased to maintain ionic homeostasis by modulating the expression of Na+ transporter genes (MhCHX15 and MhSOS1) and K+ transporter genes (MhHKT1;1, MhNHX1 and MhSKOR1). Moreover, the expression of H+-ATPase genes (MhAHA2 and MhAHA8) was increased to reduce the oxidative damage caused by saline-alkali stress. In summary, MhZEP acted as an essential role in plant resistance to saline-alkali stress, which lays the foundation for further studies on its function in apple.

Loss-of-function missense variant of AKAP4 induced male infertility through reduced interaction with QRICH2 during sperm flagella development.

Sperm fibrous sheath (FS) is closely related to sperm maturation, capacitation and motility, and A-kinase anchor protein 4 (AKAP4) is the most abundant protein in sperm FS. Previous studies found incomplete sperm FSs and abnormal flagella in Akap4 knockout mice. Meanwhile, it was reported that the partial deletion in AKAP4 is highly relevant to the dysplasia of the FS in an infertile man, and so far, there is no report about male infertility caused by hemizygous AKAP4 variant. Furthermore, the specific mechanisms of how the variant is relevant to the phenotype remain elusive. In this study, we investigated three multiple morphological abnormalities of the sperm flagella-affected men from three independent families (including one consanguine family) carried hemizygous c.C1285T variant in AKAP4. The patients carried this variant, which showed dysplastic sperm FS, and the protein expression of AKAP4 was decreased in flagella, which was further confirmed in HEK-293T cells in vitro. In addition, the co-localization and interaction between AKAP4 and glutamine-rich protein 2 (QRICH2) on the molecular level were identified by immunofluorescence and co-immunoprecipitation (CO-IP). The hemizygous c.1285C > T variant in AKAP4 induced decreased protein expression of QRICH2 in spermatozoa. These results suggested that the normal expression of AKAP4 is required for maintaining the expression of QRICH2 and the decreased protein expression of AKAP4 and QRICH2，as well as the interaction between them induced by the hemizygous variant of AKAP4 caused dysplastic fibrous sheath, which eventually led to reduced sperm motility and male infertility.

Transcriptomic analysis reveals GA3 is involved in regulating flavonoid metabolism in grape development for facility cultivation.

Gibberellin, as one of the pivotal plant growth regulators, can improve fruit quality by altering fruit size and secondary metabolite content. Flavonoids are the most abundant secondary metabolites in grapes, which influence the color and quality of the fruit. However, the molecular mechanism of whether and how GA3 affects flavonoid metabolism has not been reported, especially for the 'Red globe' grape with delayed cultivation in Hexi corridor. In the present study, the 'Red globe' grape grown in delayed facilities was sprayed with 20, 40, 60, 80 and 100 mg/L GA3 at berries pea size (BPS), veraison (V) and berries ripe (BR), respectively. The results showed that the berry weight, soluble sugar content and secondary metabolite content (the flavonoid content, anthocyanin content and polyphenol content) at BR under 80 mg/L GA3 treatment were remarkably increased compared with other concentration treatments. Therefore, RNA sequencing (RNA-seq) was used to analyze the differentially expressed genes (DEGS) and pathways under 80 mg/L GA3 treatment at three periods. GO analysis showed that DEGs were closely related to transporter activity, cofactor binding, photosynthetic membrane, thylakoid, ribosome biogenesis and other items. The KEGG enrichment analysis found that the DEGs were mainly involved in flavonoid biosynthesis and phenylpropanoid biosynthesis, indicating GA3 exerted an impact on the color and quality of berries through these pathways. In conclusion, GA3 significantly increased the expression of genes related to flavonoid synthesis, enhanced the production of secondary metabolites, and improved fruit quality. In addition, these findings can provide a theoretical basis for GA3 to modulate the accumulation and metabolism of flavonoids in grape fruit.

Transcription factors dealing with Iron-deficiency stress in plants: focus on the bHLH transcription factor family.

Iron (Fe), as an important micronutrient element necessary for plant growth and development, not only participates in multiple physiological and biochemical reactions in cells but also exerts a crucial role in respiration and photosynthetic electron transport. Since Fe is mainly present in the soil in the form of iron hydroxide, Fe deficiency exists universally in plants and has become an important factor triggering crop yield reduction and quality decline. It has been shown that transcription factors (TFs), as an important part of plant signaling pathways, not only coordinate the internal signals of different interaction partners during plant development, but also participate in plant responses to biological and abiotic stresses, such as Fe deficiency stress. Here, the role of bHLH transcription factors in the regulation of Fe homeostasis (mainly Fe uptake) is discussed with emphasis on the functions of MYB, WRKY and other TFs in the maintenance of Fe homeostasis. This review provides a theoretical basis for further studies on the regulation of TFs in Fe deficiency stress response.

The impact of meaning in life and professional happiness on the turnover intention of health care workers: a cross-sectional study from China.

INTRODUCTION: The turnover and shortage of health care workers (HCWs) have been a worldwide problem for healthcare organizations. The primary aim of this study was to identify the factors influencing the intention of Chinese HCWs to leave their job, especially meaning in life and professional happiness. METHODS: This observational cross-sectional study, conducted among 1125 full-time HCWs, assessed demographic variables, meaning in life, professional happiness, and turnover intention by a survey. The survey was distributed to HCWs in three tertiary hospitals. The data were analyzed by T-tests, ANOVA, Kruskal-Wallis tests and hierarchical linear regression model. RESULTS: There were statistically significant differences in turnover intention of HCWs by gender, age, role, educational level, years in practice, and number of monthly night shifts. HCWs' meaning in life and professional happiness were negatively associated with the turnover intention. Furthermore, after controlling for other factors, meaning in life explained 3.7% of the turnover intention and professional happiness explained 13.4%. CONCLUSION: In our study, positive psychological factors were related to turnover intentions. Professional happiness was the strongest predictor. Thus, health human resource managers should foster positive psychology among HCWs to reduce their turnover.

Time-Course and Tissue-Specific Molecular Responses to Acute Thermal Stress in Japanese Mantis Shrimp Oratosquilla oratoria.

Current understanding of adaptability to high temperatures is increasingly important as extreme weather events that can trigger immediate physiological stress in organisms have occurred more frequently. Here, we tracked transcriptomic responses of gills, hepatopancreas, and muscle to acute thermal exposure at 30 °C for 0.5, 6, and 12 h in an economically important crustacean, Oratosquilla oratoria, to gain a preliminary understanding of the tissue-specific and dynamic physiological regulation process under acute heat stress. The unique physiological responses of muscle, hepatopancreas, and gills to acute thermal stress were associated with protein degradation, lipid transport, and energy metabolism in O. oratoria, respectively. Functional enrichment analysis of differentially expressed transcripts and heat-responsive gene clusters revealed a biphasic protective responsiveness of O. oratoria developed from the early responses of signal transduction, immunity, and cytoskeleton reorganization to the responses dominated by protein turnover and energy metabolism at the mid-late stages under acute heat stress. Noteworthy, trend analysis revealed a consistently upregulated expression pattern of high molecular weight heat shock protein (HSP) family members (HSP60, HSP70, and HSP90) during the entire thermal exposure process, highlighting their importance for maintaining heat resistance in O. oratoria. Documenting the whole process of transcriptional responses at fine temporal resolution will contribute to a far-reaching comprehension of plastic responses to acute heat stress in crustaceans, which is critical in the context of a changing climate.

Relationship between family risk factors and adolescent mental health. / å®¶åº­é£Žé™©å› ç´ ä¸Žé’å°‘å¹´å¿ƒç†å¥åº·ä¹‹é—´çš„å ³ç³».

OBJECTIVES: Family risk factors are crucial to adolescent mental health. Few studies have investigated the complex relationship between family risk factors and adolescent mental health. This study aims to investigate the complex relationship between family cumulative risk (FCR) and adolescent mental health, and to clarify the factors contributing to adolescent mental health problems. METHODS: This study recruited 903 junior high school students and 991 senior high school students in Changsha, Hunan and was conducted through an offline computer-based questionnaire survey using the Middle School Student Mental Health Scale (MSSMHS) and the Family Cumulative Risk Questionnaire (FCRQ) to assess the mental health status and FCR factors, respectively. Statistical analyses were conducted to clarify the demographic factors influencing MSSMHS total and factor scores, and to analyze the relationship between FCRQ and MSSMHS total and factor scores. RESULTS: Females exhibited more mental health problems than males in various MSSMHS factors (all P<0.05); adolescents were prone to different mental health problems at different stages (junior high school first-grade vs. senior high school first-grade); senior high school first-grade students were more likely to experience academic pressure and maladjustment than junior high school first-grade students (P<0.01), and junior high school first-grade students were more likely to exhibit obsessive, paranoia, and hostility symptoms than senior high school first-grade students (all P<0.01); adolescents with low family intimacy and high family conflict reported more symptoms in every dimension of MSSMHS (all P<0.05); adolecents with poor parent-child separation reported higher obsessive-compulsive symptoms, interpersonal sensitivity, anxiety, academic pressure, maladjustment, emotional instability, and unbalanced mind than those with good parent-child separation (all P<0.05). CONCLUSIONS: Female, low family intimacy, high family conflict, and poor parent-child separation are risk factors of adolescent mental health problems. Higher-grade middle school students are prone to exhibit academic pressure and maladjustment, while lower-grade middle school students are prone to exhibit obsessive, paranoia, and hostility symptoms.

Molecular cloning and functional characterization of MhHEC2-like genes in Malus halliana reveals it enhances Fe (iron) deficiency tolerance.

The basic helix-loop-helix (bHLH) family, as one of the largest families of transcription factors (TFs) in plants, plays crucial roles in regulating growth, development, and abiotic stress responses. However, studies on the association of the bHLH genes with apple iron (Fe) deficiency are limited. Here, multiple bHLH genes that responded to Fe deficiency were selected for quantitative real-time PCR in Malus halliana. The results showed that the expression of HEC2-like gene exerted more values compared to other genes under Fe deficiency stress, but the mechanism by which it regulates Fe deficiency stress is unclear. Subsequently, MhHEC2-like gene (ID: 103,455,961) was cloned from M. halliana for functional identification. We found that both transgenic Arabidopsis thaliana and tobacco displayed less chlorosis and more robust growth than wild-type (WT) controls under Fe deficiency stress. At the same time, the overexpressed apple calli grew prominently larger and better under Fe deficiency compared to the wild type. On the other hand, physiological index measurements revealed that overexpressed MhHEC2-like gene enhanced tolerance to Fe deficiency stress in A. thaliana and tobacco by promoting the synthesis of photosynthetic pigments, improving antioxidant enzyme activity, and enhancing Fe reduction, and strengthened tolerance to Fe deficiency stress in apple calli by reducing pH, boosting Fe reduction, and increasing antioxidant enzyme activity. To sum up, the overexpression of MhHEC2-like gene strengthened tolerance to Fe deficiency stress in Arabidopsis thaliana, tobacco, and apple calli.

Genome-wide identification of the CER1 gene family in apple and response of MdCER1-1 to drought stress.

Plant cuticular wax was a major consideration affecting the growth and quality of plants through protecting the plant from drought and other diseases. According to existing studies, CER1, as the core enzyme encoding the synthesis of alkanes, the main component of wax, can directly affect the response of plants to stress. However, there were few studies on the related functions of CER1 in apple. In this study, three MdCER1 genes in Malus domestica were identified and named MdCER1-1, MdCER1-2, and MdCER1-3 according to their distribution on chromosomes. Then, their physicochemical properties, sequence characteristics, and expression patterns were analyzed. MdCER1-1, with the highest expression level among the three members, was screened for cloning and functional verification. Real-time fluorescence quantitative PCR (qRT-PCR) analysis also showed that drought stress could increase the expression level of MdCER1-1. The experiment of water loss showed that overexpression of MdCER1-1 could effectively prevent water loss in apple calli, and the effect was more significant under drought stress. Meanwhile, MdYPB5, MdCER3, and MdKCS1 were significantly up-regulated, which would be bound up with waxy metabolism. Gas chromatography-mass spectrometer assay of wax fraction makes known that overexpression of MdCER1-1 apparently scaled up capacity of alkanes. The enzyme activities (SOD, POD) of overexpressed apple calli increased significantly, while the contents of proline increased compared with wild-type calli. In conclusion, MdCER1-1 can resist drought stress by reducing water loss in apple calli epidermis, increasing alkanes component content, stimulating the expression of waxy related genes (MdYPB5, MdCER3, and MdKCS1), and increasing antioxidant enzyme activity, which also provided a theoretical basis for exploring the role of waxy in other stresses.

Horizontal transfer of large plasmid with type IV secretion system and mosquitocidal genomic island with excision and integration capabilities in Lysinibacillus sphaericus.

We identified a ~30-kb genomic island (named GI8) carrying the binary toxin gene operon binA/binB on both the chromosome and large pBsph plasmid in the mosquitocidal Lysinibacillus sphaericus C3-41 strain. We found that GI8 is related to the occurrence of binA/binB within L. sphaericus and displays excision and integration capability by recognizing the attB region, which consists of a 2-nt target site (AT) flanked by an 11-nt imperfect inverted repeat. pBsph and two pBsph-like plasmids (p2362 and p1593) were found to carry a type IV secretion system (T4SS) and displayed transmissibility within a narrow host range specific to L. sphaericus. GI8 can be co-transferred with pBsph as a composite element by integration into its attB site, then excised from pBsph and re-integrated into the chromosomal attB site in the new host. The potential hosts of GI8, regardless of whether they are toxic or non-toxic to mosquito larvae, share good collinearity at the chromosomal level. Data indicated that the appearance of the mosquitocidal L. sphaericus lineage was driven by horizontal transfer of the T4SS-type conjugative plasmid and GI8 with excision and specific integration capability.

Biotransformation of Betulonic Acid by the Fungus Rhizopus arrhizus CGMCC 3.868 and Antineuroinflammatory Activity of the Biotransformation Products.

Biotransformation of betulonic acid (1) by Rhizopus arrhizus CGMCC 3.868 resulted in the production of 16 new (3, 5, 6, and 9-21) and five known compounds. Structures of the new compounds were established by analysis of spectroscopic data. Hydroxylation, acetylation, oxygenation, glycosylation, and addition reactions involved the C-20-C-29 double bond. Antineuroinflammatory activities of the obtained compounds in NO production were tested in lipopolysaccharides-induced BV-2 cells. Compared with the substrate betulonic acid, biotransformation products 3, 8, 9, 14, and 21 exhibited an improved inhibitory effect, with IC50 values of 10.26, 11.09, 5.38, 1.55, and 4.69 µM, lower than that of the positive control, NG-monomethyl-l-arginine.

Effects of enzymatic hydrolysis on the allergenicity of natural cow milk based on a BALB/c mouse model.

Cow milk allergy is one of the most prevalent food allergies worldwide, particularly in infants and children. To the best of our knowledge, minimal research exists concerning the antigenicity of cow milk (CM). This study was performed to evaluate the allergenicity of enzymatically hydrolyzed cow milk (HM) in a BALB/c mouse model. The mice were randomly divided into 5 groups (n = 12/group), which were sensitized with phosphate-buffered saline, CM, and HM (Alcalase-, or Protamex-, or Flavorzyme-treated cow milk; Novo Nordisk; AT, PT, FT, respectively), respectively, using cholera toxin as adjuvant on d 0, 7, 14, 21. On d 28, the test mice were orally challenged with phosphate-buffered saline, CM, and HM (AT, PT, or FT) alone. Anaphylactic symptoms were monitored in the mice. Antibody, cytokine, histamine, and mouse mast cell protease-1 (mMCP-1) levels were measured using enzyme-linked immunosorbent assays. In addition, the numbers of T helper (Th)1 and Th2 cells, as well as the proportions of CD4+CD25+Foxp3+ Treg cells, in mouse spleens were detected using flow cytometry. Statistical significance was determined by one-way ANOVA. The results revealed significant differences between CM- and HM-challenged mice. Among these, the clinical scores of HM-challenged mice (AT, 1.50; PT, 2.00; FT, 1.92) were lower than those of CM-challenged mice (positive control, 2.83), but body weight and temperature of HM-challenged mice were higher than those of CM-challenged mice. In addition, significant reductions of allergen-specific IgE, IgG, histamine, and mMCP-1 were showed in HM-challenged mice, especially for histamine, ranging from 171.42 ng/mL to 214.94 ng/mL. Remarkable reductions of IL-4, IL-5, and IL-13 levels, as well as elevations of interferon-γ and IL-10 levels in the spleens of HM-challenged mice were also detected. Moreover, the number of Th2 cells decreased in the HM-challenged mice, to 2.36% (AT), 1.79% (PT), and 4.03% (FT), respectively, whereas the numbers of Th1 cells (AT, 6.30%; PT, 6.70%; FT, 6.56%) and the proportions of CD4+CD25+Foxp3+Tregs (AT, 8.86%; PT, 9.21%; FT, 9.16%) increased significantly. Our findings indicate that exposure to HM was sufficient to induce a shift toward a Th1 response, thereby reducing potential allergenicity. Importantly, these results will lay a theoretical foundation for the development of hypoallergenic CM products.

ADReCS-Target: target profiles for aiding drug safety research and application.

Delivering safe and effective therapeutic treatment to patients is one of the grand challenges in modern medicine. However, drug safety research has been progressing slowly in recent years, compared to other fields such as biotechnologies and precision medicine, due to the mechanistic complexity of adverse drug reactions (ADRs). To fill up this gap, we develop a new database, the Adverse Drug Reaction Classification System-Target Profile (ADReCS-Target, http://bioinf.xmu.edu.cn/ADReCS-Target), which provides comprehensive information about ADRs caused by drug interaction with protein, gene and genetic variation. In total, ADReCS-Target includes 66,573 pairwise relations, among which 1710 are protein-ADR associations, 2613 are genetic variation-ADR associations, and 63,298 are gene-ADR associations. In a case study of exploring the mechanism of rash, we find that HLAs, C1QA and APOA1 are the key gene players and thus can be potential targets (or biomarkers) in monitoring or countermining rashes. In summary, ADReCS-Target can be a useful resource for the biomedical scientific community by serving researchers in the fields of drug development, clinical pharmacology, precision medicine, and from web lab to high-throughput computational platform. Particularly, it helps to identify drug with better ADR profile and design safer drug therapy regimen.

Transcriptomic analysis reveals insights into deep-sea adaptations of the dominant species, Shinkaia crosnieri (Crustacea: Decapoda: Anomura), inhabiting both hydrothermal vents and cold seeps.

BACKGROUND: Hydrothermal vents and cold seeps are typical deep-sea chemosynthetically-driven ecosystems that allow high abundance of specialized macro-benthos. To gather knowledge about the genetic basis of adaptation to these extreme environments, species shared between different habitats, especially for the dominant species, are of particular interest. The galatheid squat lobster, Shinkaia crosnieri Baba and Williams, 1998, is one of the few dominant species inhabiting both deep-sea hydrothermal vents and cold seeps. In this study, we performed transcriptome analyses of S. crosnieri collected from the Iheya North hydrothermal vent (HV) and a cold seep in the South China Sea (CS) to provide insights into how this species has evolved to thrive in different deep-sea chemosynthetic ecosystems. RESULTS: We analyzed 5347 orthologs between HV and CS to identify genes under positive selection through the maximum likelihood approach. A total of 82 genes were identified to be positively selected and covered diverse functional categories, potentially indicating their importance for S. crosnieri to cope with environmental heterogeneity between deep-sea vents and seeps. Among 39,806 annotated unigenes, a large number of differentially expressed genes (DEGs) were identified between HV and CS, including 339 and 206 genes significantly up-regulated in HV and CS, respectively. Most of the DEGs associated with stress response and immunity were up-regulated in HV, possibly allowing S. crosnieri to increase its capability to manage more environmental stresses in the hydrothermal vents. CONCLUSIONS: We provide the first comprehensive transcriptomic resource for the deep-sea squat lobster, S. crosnieri, inhabiting both hydrothermal vents and cold seeps. A number of stress response and immune-related genes were positively selected and/or differentially expressed, potentially indicating their important roles for S. crosnieri to thrive in both deep-sea vents and cold seeps. Our results indicated that genetic adaptation of S. crosnieri to different deep-sea chemosynthetic environments might be mediated by adaptive evolution of functional genes related to stress response and immunity, and alterations in their gene expression that lead to different stress resistance. However, further work is required to test these proposed hypotheses. All results can constitute important baseline data for further studies towards elucidating the adaptive mechanisms in deep-sea crustaceans.

Profiling and targeting of cellular mitochondrial bioenergetics: inhibition of human gastric cancer cell growth by carnosine.

L-Carnosine (ß-alanyl-L-histidine) is a naturally occurring dipeptide distributed in various organs of mammalians. We previously showed that carnosine inhibited proliferation of human gastric cancer cells through targeting both mitochondrial bioenergetics and glycolysis pathway. But the mechanism underlying carnosine action on mitochondrial bioenergetics of tumor cells remains unclear. In the current study we investigated the effect of carnosine on the growth of human gastric cancer SGC-7901 cells in vitro and in vivo. We firstly showed that hydrolysis of carnosine was not a prerequisite for its anti-gastric cancer effect. Treatment of SGC-7901 cells with carnosine (20 mmol/L) significantly decreased the activities of mitochondrial respiratory chain complexes I-IV and mitochondrial ATP production, and downregulated 13 proteins involved in mitochondrial bioenergetics. Furthermore, carnosine treatment significantly suppressed the phosphorylation of Akt, while inhibition of Akt activation with GSK690693 significantly reduced the localization of prohibitin-1 (PHB-1) in the mitochondria of SGC-7901 and BGC-823 cells. In addition, we showed that silencing of PHB-1 gene with shRNA markedly reduced the mitochondrial PHB-1 in SGC-7901 cells, and significantly decreased the colony formation capacity and growth rate of the cells. In SGC-7901 cell xenograft nude mice, administration of carnosine (250 mg kg/d, ip, for 3 weeks) significantly inhibited the tumor growth and decreased the expression of mitochondrial PHB-1 in tumor tissue. Taken together, these results suggest that carnosine may act on multiple mitochondrial proteins to down-regulate mitochondrial bioenergetics and then to inhibit the growth and proliferation of SGC-7901 and BGC-823 cells.

First comprehensive analysis of lysine acetylation in Alvinocaris longirostris from the deep-sea hydrothermal vents.

BACKGROUND: Deep-sea hydrothermal vents are unique chemoautotrophic ecosystems with harsh conditions. Alvinocaris longirostris is one of the dominant crustacean species inhabiting in these extreme environments. It is significant to clarify mechanisms in their adaptation to the vents. Lysine acetylation has been known to play critical roles in the regulation of many cellular processes. However, its function in A. longirostris and even marine invertebrates remains elusive. Our study is the first, to our knowledge, to comprehensively investigate lysine acetylome in A. longirostris. RESULTS: In total, 501 unique acetylation sites from 206 proteins were identified by combination of affinity enrichment and high-sensitive-massspectrometer. It was revealed that Arg, His and Lys occurred most frequently at the + 1 position downstream of the acetylation sites, which were all alkaline amino acids and positively charged. Functional analysis revealed that the protein acetylation was involved in diverse cellular processes, such as biosynthesis of amino acids, citrate cycle, fatty acid degradation and oxidative phosphorylation. Acetylated proteins were found enriched in mitochondrion and peroxisome, and many stress response related proteins were also discovered to be acetylated, like arginine kinases, heat shock protein 70, and hemocyanins. In the two hemocyanins, nine acetylation sites were identified, among which one acetylation site was unique in A. longirostris when compared with other shallow water shrimps. Further studies are warranted to verify its function. CONCLUSION: The lysine acetylome of A. longirostris is investigated for the first time and brings new insights into the regulation function of the lysine acetylation. The results supply abundant resources for exploring the functions of acetylation in A. longirostris and other shrimps.

High Glucose Promotes Epithelial-Mesenchymal Transition of Uterus Endometrial Cancer Cells by Increasing ER/GLUT4-Mediated VEGF Secretion.

BACKGROUND/AIMS: Uterus endometrial cancer (UEC) is the common malignancy among gynecologic cancers, and most of them are type I estrogen-dependent UEC. Diabetes is well-known risk factor for the development of UEC. However, the underlying link between high glucose (HG) and the estrogen receptor in UEC remains unclear. Epithelial-mesenchymal transition (EMT) has also been shown to occur during the initiation of metastasis in cancer progression. The aim of this study was to determine the relationships and roles of HG, estrogen receptor and EMT in the growth and migration of UEC. METHODS: The expression of glucose transport protein 4 (GLUT4) in the control endometrium and UEC tissues was detected by immunohistochemistry (IHC); the cell viability and invasion were analyzed through CCK-8 and Matrigel invasion assays; the transcriptional level of EMT-related genes was evaluated through real-time PCR; and the effect of HG and / or GLUT4 on estrogen receptors, vascular endothelial growth factor (VEGF) and its receptor VEGFR was analyzed through western blotting, ELISA and flow cytometry (FCM) assay, respectively. In addition, Ishikawa-xenografted nude mice were constructed and were used to analyze the effect of estrogen and GLUT4 on the growth of UEC in vivo. RESULTS: Here, we found that exposure to HG led to a high level of viability and invasion of UEC cell lines (UECC, Ishikawa and RL95-2 cells). Compared with the normal endometrium, a higher level of GLUT4 was observed in UEC tissues. Silencing GLUT4 obviously inhibited the HG-promoted viability, invasion and expression of EMT-related genes (TWIST, SNAIL and CTNNB1) of UECC promoted by HG. Further analysis showed that HG and GLUT4 promoted the secretion of VEGF and expression of VEGFR in UECC. Treatment with HG led to the increase of estrogen receptor α (ERα) and ß (ERß) in UECC, blocking ERα or ERß resulted in the decreases in GLUT4 expression, TWIST, SNAIL and CTNNB1 transcription, and VEGF and VEGFR expression in UECC. Treatment with anti-human VEGF neutralizing antibody restricted the viability and invasion of UECC that was induced by HG and estrogen. Exposure to estrogen accelerated growth, VEGF production, and TWIST and CTNNB1 expression in UEC in Ishikawa-xenografted nude mice, and silencing GLUT4 restricted these effects. CONCLUSION: These data suggest that HG increases GLUT4 and VEGF/VEGFR expression, further promotes EMT process and accelerates the development of UEC by up-regulating ER.