RESUMO
Paphiopedilum orchids (Orchidaceae) have attracted much attention from botanists and horticulturists because of their peculiar leaves and beautiful flowers. Furthermore, the dry roots of Paphiopedilum plants have well-known medicinal uses. However, it is unknown how sensitive and plastic the root genes are to environmental changes or how these environmental changes regulate the biosynthesis of active ingredients. In this study, we chose Paphiopedilum concolor for root sequencing, as it is widely used as a parent in breeding experiments. A total of 3.77 Gb of sequence data were generated by Illumina paired-end sequencing. De novo assemblies yielded 72,952 contigs, 67,434 scaffolds, 64,304 unigenes with average lengths of 937, 1022, and 1047 bp, respectively. Based on Basic Local Alignment Search Tool with known protein sequences, 40,815 (63.5%) unigenes were annotated with an E-value cutoff of 1.0E-5. Among the unigenes, 24,605 were classified in the Gene Ontology database, 17,361 were assigned to Cluster of Orthologous Groups, and 14,170 were annotated in Kyoto Encyclopedia of Genes and Genomes. Among these annotations, over 1195 unigenes related to secondary metabolic pathways, as well as 609 unigenes involved in plant hormone synthesis and signal transduction, were identified. In addition, 5322 potential simple sequence repeats (SSRs) were identified, and 4989 primer pairs for 3975 sequences containing SSRs were obtained. This study provides valuable insights into the mechanisms of genes that regulate root growth and development and provides a comprehensive resource for genes related to secondary metabolism in roots and for marker-assisted studies in Paphiopedilum.
Assuntos
Repetições de Microssatélites , Orchidaceae/genética , Raízes de Plantas/genética , Análise de Sequência de DNA/métodos , Mapeamento Cromossômico , Marcadores Genéticos/genética , Anotação de Sequência Molecular , Proteínas de Plantas/genética , TranscriptomaRESUMO
Zhi-Long-Huo-Xue-Tong-Yu (ZLHXTY) is a defined mixture of 5 herbs developed by Professor S.J. Yang according to the Buyang Huanwu decoction method, which has been recorded in the Yilingaicuo. This study investigated the renoprotective effects of ZLHXTY on mitochondrial dysfunction induced by diabetic kidney injury in a diabetic rat model. Diabetes was induced by a single intravenous injection of streptozotocin. Rats were daily fed either ZLHXTY or vehicle beginning in the 1st week after injection. Levels of mitofusin 2 (mfn2), dynamin-related protein 1 (Drp1), caspase-9, and rho-associated, coiled-coil-containing protein kinase 1 (ROCK1) were detected using Western blotting. Levels of intracellular calcium and adenosine triphosphate (ATP) were examined using an enzyme-linked immunosorbent assay. An electron microscopic examination of kidney tissue was performed. The levels of mfn2 and ATP in the diabetes and ZLHXTY groups decreased from the 4th week after modeling. The expression levels of Drp1, ROCK1, and caspase-9 increased in the diabetes group but decreased in the ZLHXTY group from the 4th week after modeling. Compared with the diabetes group, ZLHXTY treatment decreased the mesangial expansion index and proteinuria levels, and improved the pathological changes typical of diabetic kidney injury. Furthermore, ZLHXTY treatment inhibited the activation of ROCK1 and expression of Drp1 and caspase-9, but did not affect the expression of mfn2. This study indicates that ZLHXTY treatment could protect kidney tissue from diabetic injury through the ROCK1 pathway response to mitochondrial dysfunction induced by diabetes.
Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Hipoglicemiantes/farmacologia , Mitocôndrias/efeitos dos fármacos , Dinâmica Mitocondrial , Quinases Associadas a rho/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Cálcio/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Medicamentos de Ervas Chinesas/uso terapêutico , Dinaminas/genética , Dinaminas/metabolismo , GTP Fosfo-Hidrolases , Hipoglicemiantes/uso terapêutico , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
We examined the expression of ezrin and moesin in laryngeal squamous cell carcinoma (LSCC) and their correlation with patient clinicopathological characteristics and overall survival. Immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) for ezrin and moesin were applied to 60 carcinoma tissues, adjacent normal tissues, and 33 metastatic lymph nodes. Survival functions were estimated using the Kaplan-Meier method and compared by the log-rank test. RT-PCR demonstrated that the intensity ratios of ezrin and moesin to ß-actin were higher in LSCC than in adjacent normal mucous membrane (P < 0.05). Furthermore, intensity ratios were higher in cervical metastatic lymph nodes than in LSCC (P < 0.05). Immunohistochemical staining showed that ezrin and moesin were well distributed in the cell membrane and cytoplasm. Expression was significantly different between LSCC and adjacent normal tissues (P < 0.05); moreover, expression in the cervical metastatic lymph nodes was higher than in LSCC (P < 0.05). Expression of ezrin and moesin was significantly related to clinical stage, T stage, and cervical lymph node metastasis (P < 0.05), except that moesin showed no significant relationship with clinical stage (P > 0.05). Patients with negative ezrin and moesin expression had a significantly longer overall survival time compared to patients with moderate and intense ezrin and moesin expression (P < 0.001, P < 0.05). Ezrin and moesin expression is related to LSCC invasion and metastasis, and may be important molecular markers for predicting prognosis and therapeutic targets in LSCC patients.
Assuntos
Carcinoma de Células Escamosas/patologia , Proteínas do Citoesqueleto/metabolismo , Neoplasias Laríngeas/patologia , Proteínas dos Microfilamentos/metabolismo , Invasividade Neoplásica , Metástase Neoplásica , Carcinoma de Células Escamosas/metabolismo , Proteínas do Citoesqueleto/genética , Feminino , Humanos , Neoplasias Laríngeas/metabolismo , Masculino , Proteínas dos Microfilamentos/genética , Pessoa de Meia-Idade , Prognóstico , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Aralia elata is an important medicinal plant in China; it produces large amounts of oleanane type triterpene saponins. A full-length cDNA encoding ß-amyrin synthase (designated as AeAS) was isolated from young leaves of A. elata by reverse transcription-PCR. The full-length cDNA of AeAS was found to have a 2292-bp open reading frame, encoding a protein with 763 amino acid residues. The deduced amino acid sequence of AeAS showed the highest identity (97%) to Panax ginseng ß-amyrin synthase. When AeAS cDNA was expressed in Escherichia coli, an 87.8-kDa recombinant protein was detected by SDS-PAGE and Western blotting. The sequence was also heterologously expressed in the yeast Pichia pastoris, and production of ß-amyrin was detected by HPLC. Tissue expression pattern analysis by real-time reverse transcription-PCR revealed that AeAS is strongly expressed in leaves and stems, and weakly expressed in roots and flowers.