RESUMO
RAD23 (RADIATION SENSITIVE23) proteins are a group of UBL-UBA (ubiquitin-like-ubiquitin-associated) proteins that shuttle ubiquitylated proteins to the 26S proteasome for breakdown. Drought stress is a major environmental constraint that limits plant growth and production, but whether RAD23 proteins are involved in this process is unclear. Here, we demonstrated that a shuttle protein, MdRAD23D1, mediated drought response in apple plants (Malus domestica). MdRAD23D1 levels increased under drought stress, and its suppression resulted in decreased stress tolerance in apple plants. Through in vitro and in vivo assays, we demonstrated that MdRAD23D1 interacted with a proline-rich protein MdPRP6, resulting in the degradation of MdPRP6 by the 26S proteasome. And MdRAD23D1 accelerated the degradation of MdPRP6 under drought stress. Suppression of MdPRP6 resulted in enhanced drought tolerance in apple plants, mainly because the free proline accumulation is changed. And the free proline is also involved in MdRAD23D1-mediated drought response. Taken together, these findings demonstrated that MdRAD23D1 and MdPRP6 oppositely regulated drought response. MdRAD23D1 levels increased under drought, accelerating the degradation of MdPRP6. MdPRP6 negatively regulated drought response, probably by regulating proline accumulation. Thus, "MdRAD23D1-MdPRP6" conferred drought stress tolerance in apple plants.
Assuntos
Malus , Ubiquitina , Ubiquitina/metabolismo , Proteínas de Transporte , Malus/genética , Proteínas de Plantas/genética , Secas , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico , Plantas Geneticamente Modificadas/metabolismoRESUMO
Erlotinib is a highly specific and reversible epidermal growth factor receptor tyrosine kinase inhibitor for the targeted therapy of non-small-cell lung cancer (NSCLC) However, the efficacy of erlotinib is limited because the development of drug resistance during chemotherapy. Indoleamine 2,3-dioxygenase-1 (IDO1) is a rate-limiting tryptophan catabolic enzyme that is activated in many human cancers. In this study, we designed a series of erlotinib-based 1,2,3-triazole compounds by combining erlotinib with phenyl or benzyl azide. Attentive FP prediction model was used to predict the bioactivity of those compounds. We discovered that most of the erlotinib-based 1,2,3-triazole compounds are capable of suppressing IDO1 activities in vitro experiments. Among them, compound 14b (IC50 = 0.59 ± 0.05 µM) had the strongest inhibitory effect on IDO1. In addition, compound 14b significantly inhibited tumor growth comparable to the antitumor activity of erlotinib and the IDO1 inhibitor epacadostat in murine tumor models.
RESUMO
Indoleamine 2,3-dioxygenase 1 (IDO1) plays a predominant role in cancer immunotherapy which catalyzes the initial and rate limiting steps of the kynurenine pathway as a key enzyme. To explore novel IDO1 inhibitors, five derivatives of erlotinib-linked 1,2,3-triazole compounds were designed by using a structure-based drug design strategy. Drug-target interactions (DTI) were predicted by DeePurpose, an easy-to-use deep learning library that contains more than 50 algorithms. The DTI prediction results suggested that the designed molecules have potential inhibitory activities for IDO1. Chemical syntheses and bioassays showed that the compounds exhibited remarkable inhibitory activities against IDO1, among them, compound e was the most potent with an IC50 value of 0.32 ± 0.07 µM in the Hela cell assay. The docking model and ADME analysis exhibited that the effective interactions of these compounds with heme iron and better drug-likeness ensured the IDO1 inhibitory activities. The studies suggested that compound e was a novel and interesting IDO1 inhibitor for further development.
RESUMO
Microsporidia are a group of unicellular and opportunistic intestinal parasites in which Enterocytozoon bieneusi is a frequent species causing microsporidial infections in humans. Many domesticated and wild animals have been shown to be hosts of E. bieneusi and other microsporidia. The role of hedgehogs in the ecology of microsporidia is unclear; therefore, we investigated the prevalence and genetic diversity of E. bieneusi, Cryptosporidium, and Blastocystis spp. in hedgehogs (Erinaceus amurensis) collected from Hubei Province in Central China. PCR amplification of the internal transcribed spacer region of the ribosomal DNA indicated that 9.8% (4/41) hedgehogs were positive to E. bieneusi, but none (0/41) was positive to Cryptosporidium and Blastocystis spp. Phylogenetic analysis showed the strains detected from the hedgehogs belong to four novel genotypes (EA1-EA4), which were most closely related to type IV of group 1c. This study demonstrated that hedgehogs are hosts of E. bieneusi and may play a role in the transmission of E. bieneusi to humans in the process of being caught and slaughtered.
Assuntos
Enterocytozoon/isolamento & purificação , Ouriços/microbiologia , Microsporidiose/veterinária , Animais , Blastocystis , China/epidemiologia , Cryptosporidium , DNA Espaçador Ribossômico , Enterocytozoon/genética , Prevalência , Análise de Sequência de DNARESUMO
Tickborne intracellular bacterial pathogens including Anaplasma, Coxiella burnetti, Ehrlichia, and Rickettsia cause emerging infectious diseases worldwide. PCR was used to amplify the genes of these pathogens in Haemaphysalis flava ticks collected from hedgehogs in Central China. Among 125 samples including 20 egg batches, 24 engorged females, and 81 molted male and female adult ticks, the DNA sequences and phylogenetic analysis showed that the minimum infection rate of the ticks was 4% (5/125) for A. bovis, 3.2% (4/125) for C. burnetti, 9.6%, (12/125) for E. ewingii, and 5.6% for Rickettsia including R.japonica (3.2%, 4/125) and R. raoultii (2.4%, 3/125), respectively. The prevalence of these pathogens was significantly higher in dead engorged females (83.3%, 20/24) than in eggs (5%, 1/20) and molted ticks (8.6%, 7/81). Our study indicated that H. flava ticks could be infected with multiple species of tickborne pathogens including Anaplasma, C. burnetti, Ehrlichia, and Rickettsia in Central China, and the prevalence of these pathogens was reduced during transovarial and transstadial transmission in ticks, suggesting that ticks may not be real reservoirs but only vectors for these tickborne pathogens.
RESUMO
The emerging coronavirus diseases such as COVID-19, MERS, and SARS indicated that animal coronaviruses (CoVs) spillover to humans are a huge threat to public health. Therefore, we needed to understand the CoVs carried by various animals. Wild hedgehogs were collected from rural areas in Wuhan and Xianning cities in Hubei Province for analysis of CoVs. PCR results showed that 5 out of 51 (9.8%) hedgehogs (Erinaceus amurensis) were positive to CoVs in Hubei Province with 3 samples from Wuhan City and 2 samples from Xianning City. Phylogenetic analysis based on the partial sequence of RNA-dependent RNA polymerase showed that the CoVs from hedgehogs are classified into Merbecovirus of the genus Betacoronavirus; the hedgehog CoVs formed a phylogenetic sister cluster with human MERS-CoVs and bat MERS-related CoVs. Among the 12 most critical residues of receptor binding domain in MERS-CoV for binding human Dipeptidyl peptidase 4, 3 residuals were conserved between the hedgehog MERS-related CoV obtained in this study and the human MERS-CoV. We concluded that hedgehogs from Hubei Province carried MERS-related CoVs, indicating that hedgehogs might be important in the evolution and transmission of MERS-CoVs, and continuous surveillance of CoVs in hedgehogs was important.
RESUMO
Coxiella burnetii is the causative agent of query fever (Q fever), and distributes broadly in environment. Livestock are identified as main reservoirs, which may infect people through their contaminative urine, feces, milk, and birth products. Wild animals can also be the potential carriers and transmitters of C. burnetii. To understand the geographic distribution and host species of C. burnetii in China, we investigated the prevalence of C. burnetii in hedgehogs (Erinaceus amurensis) in Hubei Province. Hedgehogs were tested for C. burnetii with PCR targeting three genes (com1, rrs, and icd) followed by multispacer sequence typing (MST). We found that 12.2% (5/41) hedgehogs were PCR positive for C. burnetii. MST revealed presence of two novel genotypes and phylogenetic analysis revealed that the strains were similar to a group of isolates from chronic Q fever patients and mammals. This study showed that C. burnetii are highly prevalent in hedgehogs in Hubei Province in central China, suggesting that hedgehogs may play an important role in the ecology and transmission of C. burnetii to humans because it is captured and used as traditional medicine in China.
Assuntos
Coxiella burnetii/isolamento & purificação , Ouriços/microbiologia , Febre Q/veterinária , Animais , China/epidemiologia , Coxiella burnetii/classificação , Coxiella burnetii/genética , DNA Bacteriano , Genótipo , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Febre Q/epidemiologiaRESUMO
Background:Leptospira is the causative agent of leptospirosis, a zoonotic disease of global importance. To have a better understanding on the host species of Leptospira, we investigated the prevalence of Leptospira species in hedgehogs in Central China. Materials and Methods: Hedgehogs were captured in Hubei Province, China in May and October, 2018. Total DNA was extracted from the kidney tissues of hedgehogs for determining the Leptospira species by PCR amplification of the rrs2, secY, and flaB genes with genus-specific primers. Results: PCR amplification indicated that the positive rate of hedgehogs to the rrs2, secY, and flaB genes were 19.5% (8/41), 12.2% (5/41), and 9.8% (4/41), respectively. The homology of the partial sequence of rrs2, secY, and flaB genes were 99.0-100% among the Leptospira strains from hedgehogs. Phylogenetic analysis revealed that Leptospira species detected in this study clustered together with Leptospira interrogans.Conclusions: We detected L. interrogans from hedgehogs in Central China, suggesting hedgehogs are the hosts of L. interrogans.
Assuntos
Ouriços/microbiologia , Leptospira interrogans/isolamento & purificação , Leptospirose/veterinária , Animais , China/epidemiologia , DNA Bacteriano/genética , Rim/microbiologia , Leptospira interrogans/genética , Leptospirose/epidemiologia , Leptospirose/microbiologia , Filogenia , PrevalênciaRESUMO
Conventional treatment fails to completely eliminate highly invasive breast cancer cells, and most surviving breast cancer cells tend to reproliferate and metastasize by forming vasculogenic mimicry (VM) channels. Thus, a type of targeted liposomes was developed by modification with arginine8-glycine-aspartic acid (R8GD) to encapsulate daunorubicin and emodin separately. A combination of the two targeted liposomes was then developed to destroy VM channels and inhibit tumour metastasis. MDA-MB-435S cells, a highly invasive breast cancer, were then evaluated in vitro and in mice. The experiments indicated that R8GD modified daunorubicin liposomes plus R8GD modified emodin liposomes had small particle size, uniform particle size distribution and high drug encapsulation rate. The combination of the two targeted liposomes exerted strong toxicity on the MDA-MB-435S cells and effectively inhibited the formation of VM channels and the metastasis of tumour cells. Action mechanism studies showed that the R8GD modified daunorubicin liposomes plus R8GD modified emodin liposomes could downregulate some metastasis-related proteins, including MMP-2, VE-cad, TGF-ß1 and HIF-1α. These studies also demonstrated that the targeted liposomes allowed the chemotherapeutic drug to selectively accumulate at tumour site, thus exhibiting a distinct antitumor effect. Therefore, the combination of targeted daunorubicin liposomes and targeted emodin liposomes can provide a potential treatment for invasive breast cancer.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Sistemas de Liberação de Medicamentos , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Neoplasias da Mama/patologia , Daunorrubicina/administração & dosagem , Emodina/administração & dosagem , Feminino , Humanos , Lipossomos , Camundongos , Invasividade Neoplásica , Tamanho da PartículaRESUMO
Bats have been identified as the hosts of hepatitis B virus (HBV) in recent years and bats HBV can infect human hepatocyte. We investigated the prevalence and genetic diversity of HBV in bats in China. In this study, a total of 197 insectivorous bats belonging to 10 bat species were captured from karst caves in Mengyin County, Shandong Province and Xianning City, Hubei Province, China. PCR amplification indicated that in total 6.6% (13/197) bats were positive to HBVs. The HBV positive rate in bats was 7.1% (9/127) and 5.7% (4/70) in Shandong Province and Hubei Province, respectively. Phylogenetic analysis indicated that HBV from the two places were in the same cluster with 90.5%-99.5% homology, but distinct from bat HBVs from other places in China and other countries. We concluded that HBV was prevalent and genetic diversified in bats, supporting the hypothesis that bats may be the origin of primate hepadnaviruses.
Assuntos
Quirópteros/virologia , Variação Genética , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Hepatite B/veterinária , Animais , China/epidemiologia , Eletroforese em Gel de Ágar/veterinária , Genótipo , Hepatite B/epidemiologia , Hepatite B/virologia , Vírus da Hepatite B/classificação , Humanos , Fígado/virologia , Filogenia , Reação em Cadeia da Polimerase/veterinária , PrevalênciaRESUMO
Presently, there are no few anticancer drugs that have been used clinically due to their poor targeting ability, short half-life period, non-selective distributions, generation of vasculogenic mimicry (VM) channels, high metastasis, and high recurrence rate. This study aimed to explore the effects of R8 modified epirubicin-dihydroartemisinin liposomes that could target non-small-cell lung cancer (NSCLC) cells, destroy VM channels, inhibit tumor metastasis, and explain the possible underlying mechanism. In vitro assays indicated that R8 modified epirubicin-dihydroartemisinin liposomes with ideal physicochemical characteristics could exhibit not only powerful cytotoxicity on A549 cells, but also the effective suppression of VM channels and tumor metastasis. Mechanistic studies manifested that R8 modified epirubicin-dihydroartemisinin liposomes could down-regulate the levels of VE-Cad, TGF-ß1, MMP-2, and HIF-1α. In vivo assays indicated that R8 modified epirubicin-dihydroartemisinin liposomes could both increase the selective accumulation of chemotherapeutic drugs at tumor sites and show a targeting conspicuous of antitumor efficacy. In conclusion, the R8 modified epirubicin-dihydroartemisinin liposomes prepared in this study provide a treatment strategy with high efficiency for NSCLC.
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Artemisininas/química , Carcinoma Pulmonar de Células não Pequenas/patologia , Epirubicina/química , Epirubicina/farmacologia , Neoplasias Pulmonares/patologia , Células A549 , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/metabolismo , Apoptose/efeitos dos fármacos , Transporte Biológico , Adesão Celular/efeitos dos fármacos , Liberação Controlada de Fármacos , Epirubicina/administração & dosagem , Epirubicina/metabolismo , Humanos , Bicamadas Lipídicas/metabolismo , Lipossomos , Camundongos , Invasividade Neoplásica , Metástase Neoplásica , Polietilenoglicóis/química , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Snakes are popular as food and traditional medicine in China. However, information about parasitic and bacterial infections in snakes from China is scarce. We investigated the prevalence of selected zoonotic agents including Cryptosporidium, Hepatozoon and Spirometra, in snakes in central China from June to October in 2018 by PCR amplification using parasite-specific primers. PCR amplification and DNA sequencing showed that 10.1% (15/149) of snakes were positive for Cryptosporidium spp., while 2.7% (4/149) were positive for Hepatozoon. Additionally, we found 36.9% (55/149) of snakes were infected with Spirometra erinaceieuropaei. The spargana burden per infected snake ranged from 1 to 26. BLAST and phylogenetic analysis of small subunit ribosomal RNA (SSU rRNA) gene and 60-kDa glycoprotein (gp60) gene showed that the parasites belonged to Cryptosporidium parvum genotype IIdA15G1, C. baileyi, C. serpentis and a Hepatozoon species. We conclude that intensively farmed snakes excrete C. parvum and C. baileyi oocysts due to ingestion of infected feeder animals, and that wild snakes in central China were commonly infected with S. erinaceieuropaei, suggesting that eating improperly cooked snakes could be risky to human health.
RESUMO
The contents of elements such as Zn, Fe, Mn, Cu, Na, Mg, K, Ca, As, Pb, Cd, Co, Cr, Sr, and Al in four sorts of Bulbus Fritillariae, namaly from Fritillaria thunbergii Miq. (Panan and Ningbo), Fritillaria thunbergii Miq. Var. chekiangensis Hsiao et K. C. Hsia and Fritillaria cirrhosa D. Don, were determined by flame atomic absorption spectrometry (FAAS) and graphite furnace atomic absorption spectrometry (GFAAS) respectively. The effect of different microwave digestion conditions on the analysis results was reviewed, and optimal condition for using atomic absorption spectrophotometry was determined. Principal component analysis (PCA) was applied taking trace elements contents as indexes. The results revealed that microwave digestion was a simple, rapid, digestion complete, and low blank value method, and the measurement result is satisfactory. The experimental results indicated that the linear relationships for different elements within the limits of working curve were good. The RSDs were all smaller than 3.97%. The addition standard rates of the procedure was between 91.0% and 108.7%. Principal component analysis has the application value of reflecting the differences in trace elements contents in various samples.
Assuntos
Medicamentos de Ervas Chinesas/análise , Fritillaria/química , Oligoelementos/análise , Micro-Ondas , Espectrofotometria AtômicaRESUMO
The PpERS1 gene, which encodes an ethylene receptor and responds to abiotic and biotic stresses, was cloned from peach (Prunus persica L. Batsch cv Okubao). The genomic DNA sequence of PpERS1 comprises seven exons which are separated by six introns, interestingly alternative splicing of the first intron produced three different PpERS1 transcripts. In addition, a 2.8-kb sequence including the promoter of PpERS1 was isolated and analyzed by placing expressing of the GUS reporter gene under its control. Several putative cis-elements were identified in the promoter of PpERS1, including two ethylene-responsive elements (EREs), five W boxes, and four putative binding sites for MYB-type transcription factors. Deletion analysis indicated the presence of an enhancer element in the PpERS1 promoter. Temporal and spatial expression analysis of the PpERS1 promoter using histochemical GUS staining showed GUS activity in all tissues examined throughout the development of transgenic tomato plants. Exposure to various stresses caused similar changes in expression patterns in peach and transgenic tomato plants. Overall, our results suggested that PpERS1 gene might play important roles in response to multiple stresses via signal transduction mediated by ethylene receptors. The characterization of the PpERS1 promoter contributes to our understanding of the transcriptional regulation of this ethylene receptor in peach.
RESUMO
Excessive extracellular matrix degradation caused by the hyperfunction of matrix metalloproteinases (MMPs) has been implicated in the failure of pressure ulcers healing. EMMPRIN, as a widely expressed protein, has emerged as an important regulator of MMP activity. We hypothesize that EMMPRIN affects the process of pressure ulcer healing by modulating MMP activity. In the rat pressure ulcer model, the expression of EMMPRIN in ulcers detected by Western blot was elevated compared with that observed in normal tissue. To investigate the role of EMMPRIN in regulating ulcer healing, specific antibodies against EMMPRIN were used via direct administration on the pressure ulcer. Local blockage of EMMPRIN resulted in a poor ulcer healing process compared with control ulcers, which was the opposite of our expectation. Furthermore, inhibiting EMMPRIN minimally impacted MMP activity. However, the collagen content in the pressure ulcer was reduced in the EMMPRIN treated group. Angiogenesis and the expression of angiogenic factors in pressure ulcers were also reduced by EMMPRIN local blockage. The results in the present study indicate a novel effect of EMMPRIN in the regulation of pressure ulcer healing by controlling the collagen contents and angiogenesis rather than MMPs activity.
Assuntos
Anticorpos/farmacologia , Proteínas Sanguíneas/antagonistas & inibidores , Úlcera por Pressão/metabolismo , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Proteínas Angiogênicas/metabolismo , Animais , Basigina/imunologia , Basigina/metabolismo , Proteínas Sanguíneas/imunologia , Proteínas Sanguíneas/metabolismo , Colágeno/metabolismo , Modelos Animais de Doenças , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Neovascularização Fisiológica , Úlcera por Pressão/imunologia , Úlcera por Pressão/patologia , Ratos Sprague-Dawley , Pele/irrigação sanguínea , Pele/imunologia , Pele/metabolismo , Pele/patologia , Fatores de TempoRESUMO
Citrus canker disease caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating biotic stresses affecting the citrus industry. Meiwa kumquat (Fortunella crassifolia) is canker-resistant, while Newhall navel orange (Citrus sinensis Osbeck) is canker-sensitive. To understand the molecular mechanisms underlying the differences in responses to Xcc, transcriptomic profiles of these two genotypes following Xcc attack were compared by using the Affymetrix citrus genome GeneChip. A total of 794 and 1324 differentially expressed genes (DEGs) were identified as canker-responsive genes in Meiwa and Newhall, respectively. Of these, 230 genes were expressed in common between both genotypes, while 564 and 1094 genes were only significantly expressed in either Meiwa or Newhall. Gene ontology (GO) annotation and Singular Enrichment Analysis (SEA) of the DEGs showed that genes related to the cell wall and polysaccharide metabolism were induced for basic defense in both Meiwa and Newhall, such as chitinase, glucanase and thaumatin-like protein. Moreover, apart from inducing basic defense, Meiwa showed specially upregulated expression of several genes involved in the response to biotic stimulus, defense response, and cation binding as comparing with Newhall. And in Newhall, abundant photosynthesis-related genes were significantly down-regulated, which may be in order to ensure the basic defense. This study revealed different molecular responses to canker disease in Meiwa and Newhall, affording insight into the response to canker and providing valuable information for the identification of potential genes for engineering canker tolerance in the future.