ABSTRACT
BACKGROUND: Painful diabetic neuropathy (PDN) is closely linked to inflammation, which has been demonstrated to be associated with pyroptosis. Emerging evidence has implicated TANK-binding kinase 1 (TBK1) in various inflammatory diseases. However, it remains unknown whether activated TBK1 causes hyperalgesia via pyroptosis. METHODS: PDN mice model of type 1 or type 2 diabetic was induced by C57BL/6J or BKS-DB mice with Lepr gene mutation. For type 2 diabetes PDN model, TBK1-siRNA, Caspase-1 inhibitor Ac-YVAD-cmk or TBK1 inhibitor amlexanox (AMX) were delivered by intrathecal injection or intragastric administration. The pain threshold and plantar skin blood perfusion were evaluated through animal experiments. The assessments of spinal cord, dorsal root ganglion, sciatic nerve, plantar skin and serum included western blotting, immunofluorescence, ELISA, and transmission electron microscopy. RESULTS: In the PDN mouse model, we found that TBK1 was significantly activated in the spinal dorsal horn (SDH) and mainly located in microglia, and intrathecal injection of chemically modified TBK1-siRNA could improve hyperalgesia. Herein, we described the mechanism that TBK1 could activate the noncanonical nuclear factor κB (NF-κB) pathway, mediate the activation of NLRP3 inflammasome, trigger microglia pyroptosis, and ultimately induce PDN, which could be reversed following TBK1-siRNA injection. We also found that systemic administration of AMX, a TBK1 inhibitor, could effectively improve peripheral nerve injury. These results revealed the key role of TBK1 in PDN and that TBK1 inhibitor AMX could be a potential strategy for treating PDN. CONCLUSIONS: Our findings revealed a novel causal role of TBK1 in pathogenesis of PDN, which raises the possibility of applying amlexanox to selectively target TBK1 as a potential therapeutic strategy for PDN.
Subject(s)
Diabetic Neuropathies , Microglia , Protein Serine-Threonine Kinases , Pyroptosis , Animals , Male , Mice , Aminopyridines/pharmacology , Aminopyridines/therapeutic use , Diabetic Neuropathies/pathology , Disease Models, Animal , Hyperalgesia/pathology , Mice, Inbred C57BL , Microglia/metabolism , Microglia/pathology , Microglia/drug effects , NF-kappa B/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/antagonists & inhibitors , Protein Serine-Threonine Kinases/metabolism , Protein Serine-Threonine Kinases/antagonists & inhibitors , Protein Serine-Threonine Kinases/genetics , Pyroptosis/drug effects , RNA, Small Interfering/metabolism , RNA, Small Interfering/geneticsABSTRACT
The Orchidaceae is a large family of perennial herbs especially noted for the exceptional diversity of specialized flowers. Elucidating the genetic regulation of flowering and seed development of orchids is an important research goal with potential utility in orchid breeding programs. Auxin Response Factor (ARF) genes encode auxin-responsive transcription factors, which are involved in the regulation of diverse morphogenetic processes, including flowering and seed development. However, limited information on the ARF gene family in the Orchidaceae is available. In this study, 112 ARF genes were identified in the genomes of 5 orchid species (Apostasia shenzhenica, Dendrobium catenatum, Phalaenopsis aphrodite, Phalaenopsis equestris and Vanilla planifolia,). These genes were grouped into 7 subfamilies based on their phylogenetic relationships. Compared with the ARF family in model plants, such as Arabidopsis thaliana and Oryza sativa, one group of ARF genes involved in pollen wall synthesis has been lost during evolution of the Orchidaceae. This loss corresponds with absence of the exine in the pollinia. Through mining of the published genomic and transcriptomic data for the 5 orchid species: the ARF genes of subfamily 4 may play an important role in flower formation and plant growth, whereas those of subfamily 3 are potentially involved in pollen wall development. the study results provide novel insights into the genetic regulation of unique morphogenetic phenomena of orchids, which lay a foundation for further analysis of the regulatory mechanisms and functions of sexual reproduction-related genes in orchids.
Subject(s)
Orchidaceae , Orchidaceae/genetics , Transcriptome , Phylogeny , Plant Breeding , Gene Expression Profiling , Gene Expression Regulation, Plant , Flowers/genetics , Flowers/metabolism , Plant Proteins/genetics , Plant Proteins/metabolismABSTRACT
Studies have shown that poly (ADP-ribose) polymerase 1 (PARP1) is involved in the pathological process of diabetes. Mitophagy is widely acknowledged to be a key regulatory process in maintaining reactive oxygen species homeostasis via lysosome degradation of damaged mitochondria. However, the regulatory role of PARP1 in mitophagy-related mitochondrial oxidative injury and progression of painful diabetic neuropathy (PDN) is unclear. In this study, we studied the in vitro and in vivo mechanisms of PARP1-mediated mitophagy blockade in a leptin gene-mutation (db/db) mouse model of PDN. Db/db mice models of PDN were established by assessing the sciatic nerve conduction velocity (SNCV), mechanical withdrawal threshold (MWT), and thermal withdrawal latency (TWL). The results showed that PARP1 activity and mitochondrial injury of dorsal root ganglion (DRG) neurons were increased, and mitophagy was impaired in PDN mice. PARP1 was found to mediate the impairment of mitophagy in DRG neurons isolated from PDN mice. PARP1 inhibitors (PJ34 or AG14361) attenuated diabetes-induced peripheral nerve hyperalgesia, restored DRG neuron mitophagy function and decreased mitochondrial oxidative injury. Mitophagy impairment induced by lysosome deacidificant (DC661) aggravated diabetes-induced DRG neuron mitochondrial oxidative stress and injury. Taken together, our data revealed that PARP1-induced defective mitophagy of DRG neurons is a key mechanism in diabetes-induced peripheral neuropathic injury. Inhibition of PARP1 and restoration of mitophagy function are potential therapeutic targets for PDN.
Subject(s)
Diabetes Mellitus , Diabetic Neuropathies , Adenosine Diphosphate/metabolism , Animals , Diabetes Mellitus/metabolism , Diabetes Mellitus/pathology , Ganglia, Spinal/metabolism , Mice , Mitophagy , RiboseABSTRACT
Current evidence suggests that hyperactivated or impaired autophagy can lead to neuronal death. The effect of local anesthetics on painful diabetic neuropathy (PDN) and the role of autophagy in the above pathological process remain unclear, warranting further studies. So, PDN models were established by assessing the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) in leptin gene-mutation (db/db) mice. Wild type (WT) and PDN mice received intrathecal 0.75% bupivacaine or/with intraperitoneal drug treatment (rapamycin or bafilomycin A1). Subsequently, the PWT and PWL were measured to assess hyperalgesia at 6 h, 24 h, 30 h, and 48 h after intrathecal bupivacaine. Also, sensory nerve conduction velocity (SNCV) and motor nerve conduction velocity (MNCV) were measured before and 48 h after intrathecal bupivacaine treatment. The spinal cord tissue of L4-L6 segments and serum were harvested to evaluate the change of autophagy, oxidative stress, oxidative injury, and apoptosis. We found that bupivacaine induced the activation of autophagy but did not affect the pain threshold, SNCV and MNCV in WT mice at predefined time points. Conversely, bupivacaine lowered autophagosome generation and degradation, slowed SNCV and aggravated spinal dorsal horn neuron oxidative injury and hyperalgesia in PDN mice. The autophagy activator (rapamycin) could decrease spinal dorsal horn neuron oxidative injury, alleviate the alterations in SNCV and hyperalgesia in bupivacaine-treated PDN mice. Meanwhile, the autophagy inhibitor (bafilomycin A1) could exacerbate spinal dorsal horn neuron oxidative injury, the alterations in SNCV and hyperalgesia in bupivacaine-treated PDN mice. Our results showed that bupivacaine could induce defective autophagy, slowed SNCV and aggravate spinal dorsal horn neuron oxidative injury and hyperalgesia in PDN mice. Restoring autophagy may represent a potential therapeutic approach against nerve injury in PDN patients with local anesthesia and analgesia.
Subject(s)
Diabetes Mellitus , Diabetic Neuropathies , Macrolides , Rats , Mice , Humans , Animals , Hyperalgesia/metabolism , Rats, Sprague-Dawley , Diabetic Neuropathies/chemically induced , Diabetic Neuropathies/drug therapy , Diabetic Neuropathies/metabolism , Bupivacaine/toxicity , Sirolimus , AutophagyABSTRACT
Background: Paphiopedilum armeniacum (P. armeniacum), an ornamental plant native to China, is known for its distinctive yellow blossoms. However, the mechanisms underlying P. armeniacum flower coloration remain unclear. Methods: We selected P. armeniacum samples from different flowering stages and conducted rigorous physicochemical analyses. The specimens were differentiated based on their chemical properties, specifically their solubilities in polar solvents. This key step enabled us to identify the main metabolite of flower color development of P. armeniacum, and to complete the identification by High-performance liquid chromatography (HPLC) based on the results. Additionally, we employed a combined approach, integrating both third-generation full-length transcriptome sequencing and second-generation high-throughput transcriptome sequencing, to comprehensively explore the molecular components involved. Results: We combined physical and chemical analysis with transcriptome sequencing to reveal that carotenoid is the main pigment of P. armeniacum flower color. Extraction colorimetric method and HPLC were used to explore the characteristics of carotenoid accumulation during flowering. We identified 28 differentially expressed carotenoid biosynthesis genes throughout the flowering process, validated their expression through fluorescence quantification, and discovered 19 potential positive regulators involved in carotenoid synthesis. Among these candidates, three RCP2 genes showed a strong potential for governing the PDS and ZDS gene families. In summary, our study elucidates the fundamental mechanisms governing carotenoid synthesis during P. armeniacum flowering, enhancing our understanding of this process and providing a foundation for future research on the molecular mechanisms driving P. armeniacum flowering.
Subject(s)
Gene Expression Profiling , Transcriptome , Transcriptome/genetics , Gene Expression Profiling/methods , Carotenoids/metabolism , Flowers/genetics , ChinaABSTRACT
Plants of the genus Narcissus are well-known for their characteristic corona morphology, which structural origins have been a bone of contention among scholars. With "Jinzhanyintai" (JZ) and "Yulinglong" (YLL)-two major close-originated cultivars of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem)-as materials, anatomic observation was made on floral organs during corona morphogenesis by dissection with hands under a stereomicroscope, paraffin section, scanning electron microscopy, and high-resolution X-ray tomography. It was uncovered that corona primordia of both cultivars appeared following the end of the differentiation of other floral organs, with differentiation sites located at the inner wall of the juncture of the base of tepals and the upper margin of the hypanthium. Affected by staminal filaments, the corona primordia of JZ experienced a three-stage differentiation process, namely blockage from the second whorl of stamens, blockage from the first whorl of stamens, and healing of corona primordia. However, the expanded spatial structure of the first whorl of petal-like stamens blocked the path of differentiation of YLL corona primordia, giving rise to slow differentiation of the corona primordia at the base of the first whorl of petal-like stamens and malformed differentiation of the corona primordia in the interval between the two whorls of petal-like stamens. Thus, a fragmented structure consisting of typical and fragmented coronas was formed. Furthermore, petal-like stamens of YLL in the lower part had a corona-like morphology. The spatio-temporal specificity of corona differentiation convincingly demonstrates that the corona is a structure independent of and different from the typical four whorls of floral organs, but also highly correlated with stamen.
ABSTRACT
Mesenchymal stem cells (MSCs) have been shown to improve cardiac electrophysiology when administered in the setting of acute myocardial infarction. However, the electrophysiological phenotype of MSCs in situ is not clear. We hypothesize that MSCs delivered intramyocardially to cryoinjured myocardium can engraft, but will not actively generate, action potentials. Cryoinjury-induced scar was created on the left ventricular epicardial surface of adult rat hearts. Within 30 min, hearts were injected with saline (sham, n = 11) or bone marrow-derived MSCs (2 × 10(6)) labeled with 1,1'-dioctadecyl-3,3,3,3'-tetramethylindocarbocyanine percholate (DiI; n = 16). At 3 wk, optical mapping and cell isolation were used to measure optical action potentials and calcium transients, respectively. Histological analysis confirmed subepicardial scar thickness and the presence of DiI-positive cells that express connexin-43. Optical action potential amplitude within the scar at MSC-positive sites (53.8 ± 14.3%) was larger compared with sites devoid of MSCs (35.3 ± 14.2%, P < 0.05) and sites within the scar of shams (33.5 ± 6.9%, P < 0.05). Evidence of simultaneous action potential upstroke, the loss of action potential activity following ablation of adjacent viable myocardium, and no rapid calcium transient response in isolated DiI+ cells suggest that the electrophysiological influence of engrafted MSCs is electrotonic. MSCs can engraft when directly injected into a cryoinjury and are associated with evidence of action potential activity. However, our results suggest that this activity is not due to generation of action potentials, but rather passive influence coupled from neighboring viable myocardium.
Subject(s)
Heart Conduction System/physiopathology , Heart Ventricles/physiopathology , Mesenchymal Stem Cell Transplantation , Myocardial Infarction/surgery , Myocardium/pathology , Voltage-Sensitive Dye Imaging , Action Potentials , Animals , Calcium Signaling , Carbocyanines , Cell Communication , Connexin 43/metabolism , Cryosurgery , Disease Models, Animal , Fluorescent Dyes , Heart Conduction System/metabolism , Heart Conduction System/pathology , Heart Ventricles/metabolism , Heart Ventricles/pathology , Male , Myocardial Infarction/diagnosis , Myocardial Infarction/etiology , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Myocardial Infarction/physiopathology , Myocardium/metabolism , Rats , Rats, Inbred Lew , Regeneration , Time FactorsABSTRACT
Src homology region 2 domain-containing phosphatase 1 (SHP-1) has been implicated as a potential cancer therapeutic target by its negative regulation of immune cell activation and the activity of the SHP-1 inhibitor sodium stibogluconate that induced IFN-gamma(+) cells for anti-tumor action. To develop more potent SHP-1-targeted anti-cancer agents, inhibitory leads were identified from a library of 34,000 drug-like compounds. Among the leads and active at low nM for recombinant SHP-1, tyrosine phosphatase inhibitor-1 (TPI-1) selectively increased SHP-1 phospho-substrates (pLck-pY394, pZap70, and pSlp76) in Jurkat T cells but had little effects on pERK1/2 or pLck-pY505 regulated by phosphatases SHP-2 or CD45, respectively. TPI-1 induced mouse splenic-IFN-gamma(+) cells in vitro, approximately 58-fold more effective than sodium stibogluconate, and increased mouse splenic-pLck-pY394 and -IFN-gamma(+) cells in vivo. TPI-1 also induced IFN-gamma(+) cells in human peripheral blood in vitro. Significantly, TPI-1 inhibited ( approximately 83%, p < 0.002) the growth of B16 melanoma tumors in mice at a tolerated oral dose in a T cell-dependent manner but had little effects on B16 cell growth in culture. TPI-1 also inhibited B16 tumor growth and prolonged tumor mice survival as a tolerated s.c. agent. TPI-1 analogs were identified with improved activities in IFN-gamma(+) cell induction and in anti-tumor actions. In particular, analog TPI-1a4 as a tolerated oral agent completely inhibited the growth of K1735 melanoma tumors and was more effective than the parental lead against MC-26 colon cancer tumors in mice. These results designate TPI-1 and the analogs as novel SHP-1 inhibitors with anti-tumor activity likely via an immune mechanism, supporting SHP-1 as a novel target for cancer treatment.
Subject(s)
Antineoplastic Agents/administration & dosage , Neoplasms, Experimental/drug therapy , Protein Tyrosine Phosphatase, Non-Receptor Type 6/antagonists & inhibitors , Administration, Oral , Animals , Blotting, Western , Drug Screening Assays, Antitumor , Female , Humans , Jurkat Cells , Lymphocyte Activation/drug effects , Mice , Mice, Inbred C57BLABSTRACT
Paeonia × lemoinei 'High Noon' is one of the most important cultivars in tree peony (Paeonia sect. Moutan), a traditional horticultural plant in China, with a re-blooming characteristic which was quite different from other cultivars. So, the genetic resources in 'High Noon' were incredibly valuable in flowering-time-modified molecular breeding in tree peony. However, the molecular mechanism underlying the floral bud formation of 'High Noon' was not clear yet. To explore the molecular mechanism in this process, the transcriptomes of three stages during floral bud development were deeply analyzed in this study. As a result, a total of 5816 differentially expressed genes (DEGs) were identified between the three developmental stages, and pathways including ''DNA replication'', ''metabolic pathways'', ''circadian rhythm'', and ''plant hormone signal transduction'' were mostly enriched in the functional enrichment and expression pattern analysis. Furthermore, a total of 584 genes related to the photoperiod pathway were further identified and a novel CO homolog gene PlCO was identified to be a stable hydrophilic protein, which contained both CCT domain and B-box domain. Over-expression of PlCO in Arabidopsis resulted in early flowering, which suggested a promotion role of flowering. The PlCO protein localized in nucleus and possessed a transcription activity ability, which implied that PlCO might function as a transcription factor. The transcriptome analysis revealed pathways involved in floral bud development in tree peony and provided new insight into the regulatory network underlying the floral bud development. The gene identification in 'High Noon' provided new valuable candidate genes for flowering-time-modified molecular breeding in tree peony.
Subject(s)
Paeonia , DNA/metabolism , Flowers , Gene Expression Profiling , Gene Expression Regulation, Plant , Paeonia/metabolism , Plant Growth Regulators/metabolism , Transcription Factors/metabolism , TranscriptomeABSTRACT
OBJECTIVE: To investigate an improved culturing method for karyotyping analysis, and increase the detection rate of cytogenetic abnormalities in patients with multiple myeloma (MM), so as to provide more powerful information for the clinical diagnosis, prognosis stratification, and individualized treatment of MM patients. METHODS: Eighty newly-diagnosed MM patients were enrolled and divided into two groups. In observation group, IL-6 (10 ng/ml) and GM-CSF (30 ng/ml) were supplemented in the culture medium, while no stimulating factor was added in control group. The samples from both groups were cultured for 72 hours under the same conditions, and their karyotypes were analyzed by G-banding. The detection rate of the cytogenetic abnormalities, as well as the corresponding characteristics were compared between the two groups. RESULTS: The detection rate of the chromosome aberrations was greatly increased in the observation group compared with the control group, the overall detection rate was 72.5% and 22.5%, respectively, as well as 80.0% and 19.2% in the subgroup of ≤60 years old, 68.0% and 28.6% in the subgroup of > 60 years old, which showed significant statistical differences (P<0.05). CONCLUSION: The modification of the culturing method with the addition of IL-6 (10 ng/ml) and GM-CSF (30 ng/ml) dual stimulating factors followed by incubation for 72 hours can effectively increase the detection rate of abnormal karyotypes in MM patients.
Subject(s)
Granulocyte-Macrophage Colony-Stimulating Factor , Multiple Myeloma , Chromosome Aberrations , Humans , Interleukin-6 , Karyotype , Karyotyping , Middle Aged , Multiple Myeloma/diagnosis , Multiple Myeloma/geneticsABSTRACT
Drought is a complicated and costly natural hazard and identification of critical drought factors is critical for modeling and forecasting of droughts and hence development of drought mitigation measures (the Standardized Precipitation-Evapotranspiration Index) in both space and time. Here we quantified relationships between drought and 23 drought factors using remote sensing data during the period of 2002-2016. Based on the Gradient Boosting Algorithm (GBM), we found that precipitation and soil moisture had relatively large contributions to droughts. During the growing season, the relative importance of Normalized Difference Water Index (NDWI-7) for SPEI3, SPEI6, SPEI9, and SPEI12 reached as high as 50%. However, during the non-growing season, the Snow Cover Fraction (SCF) had larger fractional relative importance for short-term droughts in the Inner Mongolia and the Loess Plateau which can reach as high as 10%. We also compared Extremely Randomized Trees (ERT), H2O-based Deep Learning (Model developed by H2O.deep learning in R H2O.DL), and Extreme Learning Machine (ELM) for drought prediction at various time scales, and found that the ERT model had the highest prediction performance with R2 > 0.72. Based on the Meta-Gaussian model, we quantified the probability of maize yield reduction in the North China Plain under different compound dry-hot conditions. Due to extreme drought and hot conditions, Shandong Province in North China had the highest probability of >80% of the maize yield reduction; due to the extreme hot conditions, Jiangsu Province in East China had the largest probability of >86% of the maize yield reduction.
Subject(s)
Droughts , Zea mays , China , Seasons , SoilABSTRACT
The present work is intended to investigate the morphological instability of lipid membrane induced by peroxyl radical (ROOâ¢) and the underlying mechanism. To this end, the giant unilamellar vesicle (GUV) made from phosphatidylcholine was employed as a membrane model, and the azo compounds 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN) and 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) were used as the precursors of ROOâ¢. Upon mild pyrolysis, the GUV immobilized in agarose gel was followed by conventional optical microscopy in real time, and the morphological variation was quantified by the image heterogeneity, perimeter and area all as a function of time for up to an hour. Lipid oxidation initiated from lipid phase with AMVN and from aqueous phase with AAPH led to different types of morphological changes, i.e. membrane coarsening and vesicle deformation/budding, respectively. Based on the compositional analysis of lipid oxidation products, we propose that ROO⢠as the primary radical initiator is responsible for the morphological changes of the GUV-AMVN while both ROO⢠and RO⢠are responsible for the morphological changes of the GUV-AAPH system. Lipophilic ß-carotene and amphipathic plant phenols as antioxidants are found to be able to stabilize the membrane integrity effectively, in corroboration with the proposed mechanisms for membrane destruction.
Subject(s)
Amidines , Unilamellar Liposomes , Amidines/pharmacology , Nitriles/pharmacology , Oxidation-Reduction , Peroxides , Phosphatidylcholines/chemistryABSTRACT
Selenium (Se) deficiency in soil is linked to its low content in edible crops, resulting in adverse impacts on the health of 15% of the global population. The crop mainly absorbs oxidized selenate and selenite from soil, then converts them into organic Se. However, the role of Se-oxidizing bacteria in soil Se oxidation, Se bioavailability and Se absorption into plants remains unclear. The strain Agrobacterium sp. T3F4, isolated from seleniferous soil, was able to oxidize elemental Se into selenite under pure culture conditions. The green fluorescent protein (gfp)-gene-marked strain (T3F4-GFP) and elemental Se or selenite (5 mg·kg-1) were added to pak choi (Brassica campestris ssp. chinensis) pot cultures. Observation of the fluorescence and viable counting indicated that GFP-expressing bacterial cells steadily colonized the soil in the pots and the leaves of the pak choi, reaching up to 6.6 × 106 and 2.0 × 105 CFU g-1 at 21 days post cultivation, respectively. Moreover, the total Se content (mostly organic Se) was significantly increased in the pak choi under T3F4 inoculated pot culture, with elemental Se(0) being oxidized into Se(IV), and soil Se(IV) being dissolved before being absorbed by the crop. After strain T3F4 was inoculated, no significant differences in microbial diversity were observed in the soils and roots, whereas the abundance of Rhizobium spp. was significantly increased. To our knowledge, this is the first time that Se-oxidizing Agrobacterium sp. T3F4 has been found to steadily colonize soil and plant tissues, and that its addition to soil increases the absorption of Se in plants. This study provides a potential strategy for Se biofortification.
Subject(s)
Brassica , Selenium , Soil Pollutants , Agrobacterium/genetics , Soil , Soil Pollutants/analysisABSTRACT
Double minute chromosomes (DMs) are rare in hematologic malignancies. We presented the cytogenetic characteristics and clinical features of the largest single-center cohort of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) patients with DMs. A total of 2576 AML patients and 1642 MDS patients were investigated, and 30 patients (AML = 19; MDS = 11) who had DMs were followed up. DMs were more common in primary AML (94.7%) and MDS (90.9%). Monosomal karyotypes (MK) were also the main cytogenetic characteristics, like complex karyotypes (CK). AML with myelodysplasia-related changes (AML-MRC) and MDS-refractory anemia with excess blasts (MDS-RAEB) was common in this cohort. We conclude that DMs-positive AML and DMs-positive MDS are associated with older age, complex karyotypes, monosomal karyotypes, TP53 deletion and TP53 mutations. DMs are a type of chromothripsis, which can be observed by the karyotype analysis. MYC and KMT2A were the most commonly amplified genes in DMs. Most patients with DMs presented an extremely poor prognosis.
Subject(s)
Leukemia, Myeloid, Acute , Myelodysplastic Syndromes , Aged , Humans , Karyotype , Karyotyping , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Mutation , Myelodysplastic Syndromes/diagnosis , Myelodysplastic Syndromes/genetics , Prognosis , Tumor Suppressor Protein p53/geneticsABSTRACT
Ca2+/calmodulin dependent protein kinase2α (CaMK2α) is a serine/threonine protein kinase in neurons and leads to neuronal injury when it is activated abnormally. Bupivacaine, a local anesthetic commonly used in regional nerve block, could induce neurotoxicity via apoptotic injury. Whether or not CaMK2α is involved in bupivacaine-induced neurotoxicity and it is regulated remains unclear. In this study, bupivacaine was administered for intrathecal injection in C57BL/6 mice for building vivo injury model and was used to culture human neuroblastoma (SH-SY5Y) cells for building vitro injury model. The results showed that bupivacaine induced mitochondrial oxidative stress and neurons apoptotic injury, promoted phosphorylation of CaMK2α and cAMP-response element binding protein (CREB), and elevated mitochondrial Ca2+ uniporter (MCU) expression. Furthermore, it induced CaMK2α phosphorylation at Thr286 which phosphorylated CREB at Ser133 and up-regulated MCU transcriptional expression. Inhibition of CaMK2α-MCU signaling with knock-down of CaMK2α and MCU or with inhibitors (KN93 and Ru360) significantly mitigated bupivacaine-induced neurotoxic injury. Over-expression of CaMK2α significantly enhanced above oxidative injury. Activated MCU with agonist (spermine) reversed protective effect of siCaMK2α on bupivacaine-induced mitochondrial oxidative stress. Our data revealed that CaMK2α-MCU-mitochondrial oxidative stress pathway is a major mechanism whereby bupivacaine induces neurotoxicity and inhibition of above signaling could be a therapeutic strategy in the treatment of bupivacaine-induced neurotoxicity.
Subject(s)
Bupivacaine , Neurotoxicity Syndromes , Animals , Bupivacaine/toxicity , Calcium/metabolism , Cell Line, Tumor , Mice , Mice, Inbred C57BL , Oxidative StressABSTRACT
The Himalayan-Tibetan Plateau (HTP), often known as the "Third Pole" and the "Asian Water Tower", is the source of water resources for many Asian rivers and in turn for hundreds of millions of people living downstream. The HTP has direct impacts on the establishment and maintenance of Asian monsoon, and therefore on the climate of its surrounding areas. Besides, soil moisture plays a critical role in the hydrological cycle and is a critical link between land surface and atmosphere. Hence, soil moisture was greatly emphasized by Global Climate Observing System Programme as an Essential Climate Variable. However, little is known about soil moisture changes on the HTP from a long-term perspective. By comparing remotely sensed and modelled soil moisture datasets against in-situ observations from 100 observation stations, here we find that Noah performed better than other soil moisture datasets. In past years, soil moisture first decreased and then increased obviously. In most regions on HTP, precipitation changes can be taken as the major cause behind soil moisture variations. In future, there is persistently decreasing soil moisture trend since ~2010 with a decreasing rate of -0.044â¯kg/m2/10a, -0.031â¯kg/m2/10a and -0.0p 88â¯kg/m2/10a under RCP2.6, RCP4.5 and RCP8.5 scenarios, respectively, in CMIP5 (Coupled Model Intercomparision Project Phase 5). Specifically, a sudden decrease of soil moisture with a rate of -0.372â¯kg/m2/10a can be expected after ~2080 under RCP8.5 scenario. Amplifying terrestrial aridity due to increasing precipitation but more significant increasing potential evapotranspiration potentially results in drying HTP. Potential water deficiency for Asian rivers due to drying HTP should arouse considerable concerns.
ABSTRACT
The Himalayan Tibet Plateau (HTP) is regarded as the third pole of the globe and is highly sensitive to global climate change. The hydrothermal properties of HTP greatly impacts the water cycle of the HTP and climate change in its surrounding regions. Using the NCEP-CFSR dataset, this study investigated the spatiotemporal pattern of soil moisture (SM) during different seasons considering vegetation types. The response of the evaporation fraction (EF) to SM and the impact of SM on air temperature through evapotranspiration were analyzed. Results showed that the spatial distribution of SM across the HTP was persistent during different seasons. A decreasing SM trend was observed from southeastern to northwestern HTP. Further, results of this study indicated a wetting tendency in past thirty years, espcially in desert region. In addition, the majority of the HTP regions were dominated by persistent transitional SM conditions which could be identified in the Himalayas and the southeastern HTP, whereas a persistent SM deficit in the Qaidam basin. The sensitivity of temperature response to EF was the strongest during spring and summer. Moreover, the spatial distribution of sensitivity was highly consistent with the vegetation regionalization, indicating the remarkable impact of vegetation type on the sensitivity of temperature to EF changes in summer.
ABSTRACT
OBJECTIVE: To verify the role of the newly identified mitotic regulator candidate pre-mRNA processing factor 19 (Prp19) in mitosis and to clarify its underlying mechanism. METHODS: FACS analyses with propidium iodide (PI) staining were performed to evaluate the effect of Prp19 knockdown on cell cycle distribution. To further clarify the role of Prp19 in mitosis, the effect of Prp19 depletion was monitored by time-lapse imaging of HeLa/GFP-H2B cells. Cold treatment experiment was used to examine the effect of Prp19 knockdown on the attachment of microtubules and kinetochores. To evaluate the effect of Prp19 knockdown on cell apoptosis, the control and Prp19-knockdown cells were analyzed by FACS with annexin V-FITC/-PI double staining. Furthermore, Western blot analysis of cleaved caspase-3 and PARP was also performed. RESULTS: Prp19 knockdown causesd mitotic arrest. Time-lapse imaging analysis showed that depletion of Prp19 in HeLa cells results in prometaphase arrest and chromosome misalignment. Cold treatment experiment showed that attachment between kinetochore and microtubule was impaired by Prp19 knockdown. Moreover, the depletion of Prp19 leaded to cell apoptosis in cancer cells. CONCLUSION: Prp19 is a key regulator of mitotic progression, and its inhibition may provide a new strategy for anti-cancer therapy.
Subject(s)
Prometaphase , Annexin A5 , Caspase 3 , Cell Cycle Proteins , DNA Repair Enzymes , Fluorescein-5-isothiocyanate/analogs & derivatives , HeLa Cells , Humans , Kinetochores , Microtubules , Mitosis , Nuclear Proteins , RNA Precursors , RNA Splicing FactorsABSTRACT
Assessing the concentration of potentially harmful heavy metals in the soil of urban parks is imperative in order to evaluate the potential risks to residents and tourists. To date, little research on soil pollution in China's urban parks has been conducted. To identify the concentrations and sources of heavy metals, and to assess the soil environmental quality, samples were collected from 30 urban parks located in the city of Beijing. Subsequently, the concentrations of Cu, Ni, Pb and Zn in the samples were analyzed. The investigation revealed that the accumulations of Cu and Pb were readily apparent in the soils. The integrated pollution index (IPI) of these four metals ranged from 0.97 to 9.21, with the highest IPI in the densely populated historic center district (HCD). Using multivariate statistic approaches (principal components analysis and hierarchical cluster analysis), two factors controlling the heavy metal variability were obtained, which accounted for nearly 80% of the total variance. Nickel and Zn levels were controlled by parent material in the soils, whereas Cu, Pb and, in part, Zn were accounted for mainly by anthropogenic activities. The findings presented here indicate that the location and the age of the park are important factors in determining the extent of heavy metal, particularly Cu and Pb, pollution. In addition, the accumulation of Zn did not appear to reach pollution levels, and no obvious pollution by Ni was observed in the soils of the parks in Beijing.
Subject(s)
Metals, Heavy/analysis , Soil Pollutants/analysis , China , Cities , Environmental Monitoring , RecreationABSTRACT
Drug resistance is a major obstacle in cancer treatments and diminishes the clinical efficacy of biological, cytotoxic, or targeted therapeutics. Being an antiapoptotic mediator of chemoresistance in breast and lung cancer cells, MKP1 phosphatase might be targeted for overcoming chemoresistance and improving therapeutic efficacy. In this work, tyrosine phosphatase inhibitor-3 (TPI-3) was identified as a novel small molecule inhibitor of MKP1 and was capable of sensitizing tumors to bio- and chemotherapeutics in mice as a tolerated oral agent. Effective against recombinant MKP1, TPI-3 selectively increased MKP1 phosphosubstrates in Jurkat cells and induced cell death via apoptosis at nanomolar concentrations. TPI-3 also increased MKP1 phosphosubstrates in WM9 human melanoma cells and synergized with biotherapeutic IFN alpha 2b in the growth inhibition of melanoma cells in vitro (combination index, <1). WM9 xenografts unresponsive to individual agents were significantly inhibited (62%, P = 0.001) in mice by a tolerated combination of oral TPI-3 (10 mg/kg, 5 d/wk) and IFN alpha 2b. MKP1 expression was detected in human melanoma cell lines and tissue samples at levels up to six times higher than those in normal or nonmalignant melanocytes. TPI-3 also interacted positively with chemotherapeutics, 5-fluorouracil/leucovorin, against MC-26 colon cancer cells in vitro and in mice. Altogether, our data show the preclinical activities of TPI-3 in overcoming cancer resistance to bio- and chemotherapeutics, implicate MKP1 as a drug-resistant molecule in melanoma, and support the targeting of MKP1 for improving cancer therapeutic efficacy.