RESUMEN
An approach based on the heat stress and microbial stress model of the medicinal plant Sparganium stoloniferum was proposed to elucidate the regulation and mechanism of bioactive phenol accumulation. This method integrates LC-MS/MS analysis, 16S rRNA sequencing, RT-qPCR, and molecular assays to investigate the regulation of phenolic metabolite biosynthesis in S. stoloniferum rhizome (SL) under stress. Previous research has shown that the metabolites and genes involved in phenol biosynthesis correlate to the upregulation of genes involved in plant-pathogen interactions. High-temperature and the presence of Pseudomonas bacteria were observed alongside SL growth. Under conditions of heat stress or Pseudomonas bacteria stress, both the metabolites and genes involved in phenol biosynthesis were upregulated. The regulation of phenol content and phenol biosynthesis gene expression suggests that phenol-based chemical defense of SL is stimulated under stress. Furthermore, the rapid accumulation of phenolic substances relied on the consumption of amino acids. Three defensive proteins, namely Ss4CL, SsC4H, and SsF3'5'H, were identified and verified to elucidate phenol biosynthesis in SL. Overall, this study enhances our understanding of the phenol-based chemical defense of SL, indicating that bioactive phenol substances result from SL's responses to the environment and providing new insights for growing the high-phenol-content medicinal herb SL.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Respuesta al Choque Térmico , Plantas Medicinales , Plantas Medicinales/metabolismo , Fenoles/metabolismo , Fenol/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Rizoma/microbiología , Rizoma/metabolismo , Pseudomonas/metabolismo , Pseudomonas/genética , Espectrometría de Masas en Tándem , ARN Ribosómico 16S/genéticaRESUMEN
The volatile oils are the effective components of Agastache rugosa, which are stored in the glandular scale. The leaves of pulegone-type A. rugosa were used as materials to observe the leaf morphology of A. rugosa at different growth stages, and the components of volatile oils in gland scales were detected by GC-MS. At the same time, qRT-PCR was used to determine the relative expression of key enzyme genes in the biosynthesis pathway of monoterpenes in volatile oils. The results showed that the density of A. rugosa glandular scale decreased first and then tended to be stable. With the growth of leaves, the relative content of pulegone decreased from 79.26% to 3.94%(89.97%-41.44%), while that of isomenthone increased from 2.43% to 77.87%(0.74%-51.01%), and the changes of other components were relatively insignificant. The correlation analysis between the relative content of monoterpenes and the relative expression levels of their key enzyme genes showed that there was a significant correlation between the relative content of menthone and isomenthone and the relative expression levels of pulegone reductase(PR)(r>0.6, P<0.01). To sum up, this study revealed the accumulation rules of the main components of the contents of the glandular scale of A. rugosa and the expression rules of the key enzyme genes for biosynthesis, which provided a scientific basis and data support for determining the appropriate harvesting period and quality control of the medicinal herbs. This study also initially revealed the biosynthesis mechanism of the monoterpenes mainly composed of pulegone and isomenthone in A. rugosa, laying a foundation for further research on the molecular mechanism of synthesis and accumulation of monoterpenes in A. rugosa.
Asunto(s)
Agastache , Monoterpenos Ciclohexánicos , Aceites Volátiles , Aceites Volátiles/análisis , Agastache/metabolismo , Monoterpenos/metabolismoRESUMEN
Terpenoids are important secondary metabolites of plants that possess both pharmacological activity and economic value. Terpene synthases(TPSs) are key enzymes in the synthesis process of terpenoids. In order to investigate the TPS gene family members and their potential functions in Schizonepeta tenuifolia, this study conducted a systematic analysis of the TPS gene family of S. tenuifolia based on the whole genome data of S. tenuifolia using bioinformatics methods. The results revealed 57 StTPS members identified from the genome database of S. tenuifolia. The StTPS family members encoded 285-819 amino acids, with protein molecular weights ranging from 32.75 to 94.11 kDa, all of which were hydrophilic proteins. The StTPS family members were mainly distributed in the cytoplasm and chloroplasts, exhibiting a random and uneven physical localization pattern. Phylogenetic analysis showed that the StTPS genes family were divided into six subgroups, mainly belonging to the TPS-a and TPS-b subfamilies. Promoter analysis predicted that the TPS gene family members could respond to various stressors such as light, abscisic acid, and methyl jasmonate(MeJA). Transcriptome data analysis revealed that most of the TPS genes were expressed in the roots of S. tenuifolia, and qRT-PCR analysis was conducted on genes with high expression in leaves and low expression in roots. Through the analysis of the TPS gene family of S. tenuifolia, this study identified StTPS5, StTPS18, StTPS32, and StTPS45 as potential genes involved in sesquiterpene synthesis of S. tenuifolia. StTPS45 was cloned for the construction of an prokaryotic expression vector, providing a reference for further investigation of the function and role of the TPS gene family in sesquiterpene synthesis.
Asunto(s)
Lamiaceae , Sesquiterpenos , Filogenia , Terpenos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Lamiaceae/genéticaRESUMEN
Ursolic acid (UA) exists in a variety of medicinal plants. UA exhibits antimicrobial activity against several microorganisms; however, little is known regarding the potential antifungal effect of UA on Cryptococcus neoformans (C. neoformans). The antifungal and antibiofilm activities of UA on C. neoformans H99 were evaluated in this study. Minimum inhibitory concentration (MIC) of UA against C. neoformans H99 was determined by microdilution technique, and its action mode was elucidated by clarifying the variations in cell membrane integrity, capsule, and melanin production. Moreover, the inhibition and dispersal effects of UA on biofilm formation and mature biofilms by C. neoformans H99 were evaluated using crystal violet (CV) assay, optical microscopy, field emission scanning electron microscopy and confocal laser scanning microscopy. The results indicated that the MIC value of UA against C. neoformans H99 was 0.25 mg/mL. UA disrupted the cell membrane integrity, inhibited the capsule and melanin production of C. neoformans H99 in a concentration-dependent manner. Further, UA presented the inhibitory effect on biofilm formation and dispersed mature biofilms, as well as compromised the cell membrane integrity of C. neoformans H99 cells within biofilms. Together, these results indicate that UA might be a potential therapeutic option for the treatment of C. neoformans-related infections.
Asunto(s)
Criptococosis , Cryptococcus neoformans , Antifúngicos/metabolismo , Antifúngicos/farmacología , Biopelículas , Criptococosis/tratamiento farmacológico , Cryptococcus neoformans/metabolismo , Melaninas/metabolismo , Melaninas/farmacología , Pruebas de Sensibilidad Microbiana , Triterpenos , Ácido UrsólicoRESUMEN
Hd-Zip, a unique transcription factor in plant kingdom, influences the growth, development, and secondary metabolism of plants. Hd-zip â £ is thought to play an important role in trichome development of Schizonepeta tenuifolia. This study aims to explore the functions of StHD1 and StHD8 in Hd-zip â £ subfamily in peltate glandular trichome development. To be specific, the expression patterns of the two genes and interaction between the proteins encoded by them were analyzed based on transcriptome sequencing and two-hybrid screening. The subcellular localization was performed and functions of the genes were verified in tobacco and S. tenuifolia. The results showed that StHD1 and StHD8 had high similarity to HD-Zip â £ proteins of other plants and they all had the characteristic conserved domains of HD-Zip â £ subfamily. They were located in the nucleus. The two genes mainly expressed in young tissues and spikes, and StHD1 and StHD8 proteins interacted with each other. The density and length of glandular trichomes increased significantly in tobacco plants with the overexpression of StHD1 and StHD8. Inhibiting the expression of StHD1 and StHD8 by VIGS(virus-induced gene silencing) in S. tenuifolia resulted in the reduction in the density of peltate glandular trichomes, the expression of key genes related to mono-terpene synthesis, and the relative content of limonene and pulegone, the main components of monoterpene. These results suggested that StHD1 and StHD8 of S. tenuifolia formed a complex to regulate glandular trichomes and affect the biosynthesis of monoterpenes.
Asunto(s)
Lamiaceae , Tricomas , Tricomas/genética , Tricomas/metabolismo , Lamiaceae/genética , Nicotiana/genética , Monoterpenos/metabolismo , Clonación Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
BACKGROUND: Perilla frutescens (L.) Britt is a medicinal and edible plant widely cultivated in Asia. Terpenoids, flavonoids and phenolic acids are the primary source of medicinal ingredients. Glandular trichomes with multicellular structures are known as biochemical cell factories which synthesized specialized metabolites. However, there is currently limited information regarding the site and mechanism of biosynthesis of these constituents in P. frutescens. Herein, we studied morphological features of glandular trichomes, metabolic profiling and transcriptomes through different tissues. RESULTS: Observation of light microscopy and scanning electron microscopy indicated the presence of three distinct glandular trichome types based on their morphological features: peltate, capitate, and digitiform glandular trichomes. The oil of peltate glandular trichomes, collected by custom-made micropipettes and analyzed by LC-MS and GC-MS, contained perillaketone, isoegomaketone, and egomaketone as the major constituents which are consistent with the components of leaves. Metabolomics and transcriptomics were applied to explore the bioactive constituent biosynthesis in the leaves, stem, and root of P. frutescens. Transcriptome sequencing profiles revealed differential regulation of genes related to terpenoids, flavonoids, and phenylpropanoid biosynthesis, respectively with most genes expressed highly in leaves. The genes affecting the development of trichomes were preliminarily predicted and discussed. CONCLUSIONS: The current study established the morphological and chemical characteristics of glandular trichome types of P. frutescens implying the bioactive constituents were mainly synthesized in peltate glandular trichomes. The genes related to bioactive constituents biosynthesis were explored via transcriptomes, which provided the basis for unraveling the biosynthesis of bioactive constituents in this popular medicinal plant.
Asunto(s)
Perilla frutescens/química , Tricomas/química , Regulación de la Expresión Génica de las Plantas , Microscopía Electrónica de Rastreo , Perilla frutescens/genética , Perilla frutescens/ultraestructura , Reguladores del Crecimiento de las Plantas/metabolismo , Hojas de la Planta/química , Raíces de Plantas/química , Tallos de la Planta/química , Plantas Medicinales/química , Plantas Medicinales/ultraestructura , ARN de Planta , RNA-Seq , Reacción en Cadena en Tiempo Real de la Polimerasa , Transcriptoma , Tricomas/ultraestructuraRESUMEN
To explore posttraumatic stress disorder symptoms (PTSD) after respiratory insufficiency in patients with myasthenia gravis (MG). The investigation was made with 134 adult patients with MG, after respiratory insufficiency, between January 2012 and January 2016 and had a return visit after one year. 134 patients finished this study and 69 patients (51.5%) had PTSD. Anxiety (HADS-A ≥ 8, HADS: Hospital Anxiety and Depression Scale) (OR 2.585,95% CI 1.102-6.061, p = 0.029), and depression (HADS-D ≥ 8) (OR 3.200, 95% CI 1.395-7.342, p = 0.006) were associated with greater probabilities of screening positive for PTSD. Gender, age, intubation, yearly income, marriage, inability to work, number of respiratory insufficiency episodes, education level, Mini-mental state examination (MMSE) (>20), ICU stays, having insurance, and MG-activities of daily living (ADL) (<9) were not significant predictors for PTSD. One year after a respiratory insufficiency episode, patients with PTSD experienced worse anxiety (p = 0.035), depressive disorder (p < 0.001), and 36-Item Short-Form Health Survey (SF-36) showed physical functioning (p = 0.042), role-physical (p = 0.013), social functioning (p = 0.040), and emotional-role (p = 0.034). But there were no differences in ADL, bodily pain, general health and vitality. PTSD in patients with MG is common after a respiratory insufficiency episode; anxiety and depression were both associated with greater probabilities of screening positive for PTSD.
Asunto(s)
Miastenia Gravis/terapia , Insuficiencia Respiratoria/psicología , Trastornos por Estrés Postraumático/epidemiología , Adulto , China/epidemiología , Femenino , Hospitalización , Humanos , Masculino , Persona de Mediana Edad , Miastenia Gravis/complicaciones , Insuficiencia Respiratoria/etiología , Estudios RetrospectivosRESUMEN
Based on the characteristics and ISSR molecular marker technology, the study is aimed to compare and perform genetic diversity analysis on Sparganium stoloniferum from 7 regions. Molecular identification method was established for S. stoloniferum from Hunan province. Differences among Sparganii Rhizoma samples from seven habitats were analyzed via measuring weight, length, width and thickness of them. Genetic diversity of S. stoloniferum from 7 regions was analyzed by screening out primers amplifying clear band and showing rich polymorphism, then a cultivars dendrogram was built. The target primer was screened out, and the specific band was sequenced. Nine ISSR primers were selected to amplified clear band, rich polymorphism. A total of 73 bands were amplified by nine ISSR primers selected from 27 ISSR primers. On average, each primer produced 8.0 bands. A total of 38 bands were polymorphic, which occupied 52.8% of all bands. The cultivars dendrogram showed the genetic similarity was 0.54-0.94. Genetic similarity coefficient of S. stoloniferum from Jiangsu province, Anhui province and Jiangxi province was big, indicating the differences among them were slight on genetic level. S. stoloniferum from Hunan province is quite different from samples from the other six habitats on appea-rance and genetic level. A specific band(327 bp) in S. stoloniferum from Hunan province was obtained via ISSR-857 primer, and was sequenced. According BLASTn database, there were few sequences similar to the gene fragment and had little correlation with the growth process of plant. ISSR molecular marker technology provides a new idea for the identification of S. stoloniferum. This result confirmed the particularity of S. stoloniferum from ancient Jingzhou.
Asunto(s)
Medicamentos Herbarios Chinos , Variación Genética , Polimorfismo Genético , China , Marcadores Genéticos/genética , Repeticiones de Microsatélite , FilogeniaRESUMEN
Leaves of Euryale ferox are rich in anthocyanins. Anthocyanin synthesis is one of the important branches of the flavonoid synthesis pathway, in which flavonoid 3'-hydroxylase(F3'H) can participate in the formation of important intermediate products of anthocyanin synthesis. According to the data of E. ferox transcriptome, F3'H cDNA sequence was cloned in the leaves of E. ferox and named as EfF3'H. The correlation between EfF3'H gene expression and synthesis of flavonoids was analyzed by a series of bioinforma-tics tools and qRT-PCR. Moreover, the biological function of EfF3'H was verified by the heterologous expression in yeast. Our results showed that EfF3'H comprised a 1 566 bp open reading frame which encoded a hydrophilic transmembrane protein composed of 521 amino acid residues. It was predicted to be located in the plasma membrane. Combined with predictive analysis of conserved domains, this protein belongs to the cytochrome P450(CYP450) superfamily. The qRT-PCR results revealed that the expression level of EfF3'H was significantly different among different cultivars and was highly correlated with the content of related flavonoids in the leaves. Eukaryotic expression studies showed that EfF3'H protein had the biological activity of converting kaempferol to quercetin. In this study, EfF3'H cDNA was cloned from the leaves of E. ferox for the first time, and the biological function of the protein was verified. It provi-ded a scientific basis for further utilizing the leaves of E. ferox and laid a foundation for the further analysis of the biosynthesis pathway of flavonoids in medicinal plants.
Asunto(s)
Antocianinas , Proteínas de Plantas , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , TranscriptomaRESUMEN
Depression, anxiety and posttraumatic stress disorder (PTSD) are common complications of cerebrovascular diseases. However, they were seldom explored in Moyamoya Disease (MMD) survivors. In this study, we measured the prevalence of depression, anxiety and PTSD in MMD survivors. We evaluated the association of mental disorders with neurological disability and cognitive impairment, and further find out the independent protective and risk factors of neurological disability and cognitive impairment. In MMD survivors, the prevalence of these three mental disorders is high, 46.7% for depression, 50% for anxiety and 47.5% for PTSD. Anxiety and PTSD were significantly associated with more severe neurological disability (p = 0.039 and < 0.001); depression and anxiety were significantly associated with greater cognitive deficiency (p = 0.004 and 0.002). We further found PTSD was the only risk factor associated with neurological disability, and the corresponding odds ratio (OR) and 95% confidence interval (CI) was 81.74 (9.91-674.17); depression and anxiety were risk factors associated with cognitive impairment, and the corresponding OR and 95%CI were 2.73 (1.10-6.81) and 3.37 (1.29-8.78). Therefore, these three mental disorders were associated with more severe neurological disability and greater cognitive deficiency in MMD survivors.
Asunto(s)
Trastornos de Ansiedad/epidemiología , Disfunción Cognitiva/epidemiología , Trastorno Depresivo/epidemiología , Enfermedad de Moyamoya/epidemiología , Trastornos por Estrés Postraumático/epidemiología , Adulto , Comorbilidad , Femenino , Humanos , Masculino , Índice de Severidad de la Enfermedad , Sobrevivientes/estadística & datos numéricosRESUMEN
The etiology and pathogenesis of Parkinson's disease (PD) are complicated and have not been fully elucidated, but an important association has been identified between inflammation and PD. In this study, we investigated the role of the nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain-containing (NLRP) 3 inflammasome, consisting of NLRP3, caspase-1 and cytokines of the IL-1 family, in lipopolysaccharide (LPS)-induced and 6-hydroxydopamine (6-OHDA)-induced PD rats. Microinjection of different doses of caspase-1 inhibitor (Ac-YVAD-CMK, 300 or 1200 ng/rat) was performed for seven consecutive days. Then, rotational behavior, the number of dopamine (DA) neurons in the substantia nigra pars compacta (SNc), and the mRNA and protein expression levels of NLRP3 inflammasome components were measured 14 days after the microinjection setup was established. Results showed that high mRNA and protein expression levels of NLRP3 inflammasome components were observed in the injected side of the LPS- and 6-OHDA-induced PD rats; Ac-YVAD-CMK inhibited the mRNA and protein expression of NLRP3 inflammasome components in both LPS- and 6-OHDA-induced PD rats. Moreover, the number of rotations was significantly decreased, and the number of DA neurons in the SNc improved. Our data indicate that the NLRP3 inflammasome participates in the pathogenesis of PD and that inhibiting the downstream pathway of the NLRP3/caspase-1/IL-1ß axis can alleviate the occurrence of PD symptoms, providing a new basis for the prevention and treatment of PD.
Asunto(s)
Inflamasomas/biosíntesis , Proteína con Dominio Pirina 3 de la Familia NLR/biosíntesis , Trastornos Parkinsonianos/metabolismo , Trastornos Parkinsonianos/patología , Animales , Masculino , Oxidopamina/toxicidad , Trastornos Parkinsonianos/inducido químicamente , Distribución Aleatoria , Ratas , Ratas Sprague-DawleyRESUMEN
PURPOSE: To compare the post-thymectomy prognosis in different conditions of myasthenia gravis (MG) patients with thymus hyperplasia. MATERIALS AND METHODS: Collecting medical record and carrying out the follow-up study of 123 myasthenia gravis patients with thymus hyperplasia who have underwent thymectomy during the period between 2003 and 2013. Dividing into different groups based on gender, age of onset, duration of disease and Myasthenia Gravis Association of America (MGFA) clinical classification to analyze different prognosis in different groups. RESULTS: Complete stable remission (CSR) was achieved in 71 of 123 patients (59.5%). There is no gender-related difference in achieving CSR. Patients with early onset of MG (≤40 years old) or disease duration less than 12 months had significantly higher CSR rates than those with late onset of MG (>40 years old) or disease duration more than 12 months respectively, while no difference was found in remission rate between MGFA clinical classification I and MGFA II. CONCLUSION: Myasthenia gravis patients with thymus hyperplasia who had thymectomy are proved to possess greater chance of achieving CSR. The onset age of disease and duration are the prognostic factors.
Asunto(s)
Miastenia Gravis/complicaciones , Miastenia Gravis/cirugía , Timectomía/métodos , Hiperplasia del Timo/complicaciones , Hiperplasia del Timo/cirugía , Resultado del Tratamiento , Adolescente , Adulto , Anciano , Autoanticuerpos/metabolismo , Femenino , Estudios de Seguimiento , Humanos , Inmunoglobulinas Intravenosas/uso terapéutico , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Miastenia Gravis/diagnóstico por imagen , Miastenia Gravis/terapia , Pronóstico , Modelos de Riesgos Proporcionales , Receptores Colinérgicos/inmunología , Estudios Retrospectivos , Hiperplasia del Timo/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Adulto JovenRESUMEN
OBJECTIVE: To analyze the mortality and potential risk factors for death in myasthenia gravis (MG) patients. MATERIALS AND METHODS: A total of 2195 adult patients with MG (aged older than 18 years) diagnosed during the period between 2003 and 2013 were followed-up and retrospectively reviewed. RESULTS: During the 10-year follow-up, 129 patients died and the total mortality rate was 5.88%. The risk factors associated with MG-related deaths were duration of the disease, occurrence of myasthenic crisis, severity of disease that included the Myasthenia Gravis Foundation of America (MGFA) grade III and IV at onset, elevation of acetylcholine receptor antibody (AchR-abs) titers, presence of thymic pathology, and failure of administrating immunosuppressants (P < 0.05). In addition, the non-MG related factors, including the history of preceding strokes, and the presence of chronic obstructive pulmonary disease (COPD), diabetes mellitus, atrial fibrillation, hyperlipidemia, myocardial infarction, and malignant tumors, were closely linked with death in the MG population (the hazard ratios [HRs] were 3.251, 4.173, 3.738, 3.886, 1.945, 2.177, and 14.7, respectively; P< 0.05). CONCLUSIONS: The severity of disease at entry, presence of AchRabs, thymic pathology, and duration of the disease predict a higher risk for death. Systemic illnesses including stroke, COPD, diabetes mellitus, atrial fibrillation, hyperlipidemia, myocardial infarction, and malignant tumor, which may also increase the risk of death, should be carefully monitored and managed.
Asunto(s)
Miastenia Gravis/complicaciones , Miastenia Gravis/mortalidad , Adolescente , Adulto , Distribución por Edad , Anciano , Fibrilación Atrial/etiología , Autoanticuerpos/sangre , Diabetes Mellitus/etiología , Femenino , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Miastenia Gravis/epidemiología , Miastenia Gravis/terapia , Enfermedad Pulmonar Obstructiva Crónica/etiología , Receptores Colinérgicos/inmunología , Estudios Retrospectivos , Factores de Riesgo , Estadísticas no Paramétricas , Accidente Cerebrovascular/etiología , Timectomía , Timo/patología , Timo/cirugía , Adulto JovenRESUMEN
A gas chromatography-mass spectrometry(GC-MS)method was established for the analysis of volatile components in Mentha haplocalyx, and seven principal components were quantified by gas chromatography(GC). Based on these analyses, the differences of volatile components in M. haplocalyx from Jiangsu, Anhui and other regions were compared. The results showed that the volatile oil of M. haplocalyx was divided into four chemical typesï¼menthol-menthone type, pulegone-menthone type, piperitone-menthol type, piperitone epoxide type, and menthol-menthone type was the principal type. Menthol was the highest and pulegone was the lowest. The differences of M. haplocalyx from Anhui and other regions were obvious. The major volatile components and the differences of M. haplocalyx from different regions were confirmed and a quantitative method was established for the determination of volatile components, which provided the basis for improving the quality standard of M. haplocalyx.
Asunto(s)
Medicamentos Herbarios Chinos/normas , Mentha/química , Aceites Volátiles/química , Cromatografía de Gases y Espectrometría de Masas , Aceites de Plantas/química , Plantas Medicinales/químicaRESUMEN
Fuzi is a medicine used for rescuing from collapse by restoring yang as well as a famous toxic traditional Chinese medicine. In order to ensure the efficacy and safe medication, Fuzi has mostly been applied after being processed. There have been different Fuzi processing methods recorded by doctors of previous generations. Besides, there have also been differences in Fuzi processing methods recorded in modern pharmacopeia and ancient medical books. In this study, the authors traced back to medical books between the Han Dynasty and the period of Republic of China, and summarized Fuzi processing methods collected in ancient and modern literatures. According to the results, Fuzi processing methods and using methods have changed along with the evolution of dynasties, with differences in ancient and modern processing methods. Before the Tang Dynasty, Fuzi had been mostly processed and soaked. From Tang to Ming Dynasties, Fuzi had been mostly processed, soaked and stir-fried. During the Qing Dynasty, Fuzi had been mostly soaked and boiled. In the modem times, Fuzi is mostly processed by being boiled and soaked. Before the Tang Dynasty, a whole piece of Fuzi herbs or their fragments had been applied in medicines; Whereas their fragments are primarily used in the modern times. Because different processing methods have great impacts on the toxicity of Fuzi, it is suggested to study Fuzi processing methods.
Asunto(s)
Química Farmacéutica/historia , Química Farmacéutica/métodos , Medicamentos Herbarios Chinos/química , Extractos Vegetales/química , China , Diterpenos , Historia del Siglo XV , Historia del Siglo XVI , Historia del Siglo XVII , Historia del Siglo XVIII , Historia del Siglo XIX , Historia Antigua , Historia Medieval , Humanos , Medicina Tradicional China/historia , Medicina Tradicional China/métodosRESUMEN
This study investigates the impact of methyl jasmonate (MeJA) on the volatile oil composition of Schizonepeta tenuifolia and elucidates the function of the StTPS45 gene, a key player in terpenoid biosynthesis. The effect of different concentrations of MeJA (0, 50, 100, 200, and 300 µmol/L) on the growth of S. tenuifolia adventitious bud clusters was analyzed over a 20 d period. Using gas chromatography-mass spectrometry (GC-MS), 17 compounds were identified from the adventitious bud clusters of S. tenuifolia. Significant changes in the levels of major monoterpenes, including increased contents of (+)-limonene and (+)-menthone, were observed, particularly at higher concentrations of MeJA. Analysis of transcriptome data from three groups treated with 0, 100, and 300 µmol/L MeJA revealed significant changes in the gene expression profiles following MeJA treatment. At 100 µmol/L MeJA, most terpene synthase (TPS) genes were overexpressed. Additionally, gene expression and functional predictions suggested that StTPS45 acts as germacrene D synthase. Therefore, StTPS45 was cloned and expressed in Escherichia coli, and enzyme activity assays confirmed its function as a germacrene D synthase. Molecular docking and structural prediction of StTPS45 further suggested specific interactions with farnesyl diphosphate (FPP), aligning with its role in the terpenoid synthesis pathway. These findings provide valuable insights into the modulation of secondary metabolite pathways by jasmonate signaling and underscore the potential of genetic engineering approaches to enhance the production of specific terpenoids in medicinal plants.
RESUMEN
Menthone-type monoterpenes are the main active ingredients of Schizonepeta tenuifolia Briq. Previous studies have indicated that light intensity influences the synthesis of menthone-type monoterpenes in S. tenuifolia, but the mechanism remains unclear. WRKY transcription factors play a crucial role in plant metabolism, yet their regulatory mechanisms in S. tenuifolia are not well understood. In this study, transcriptome data of S. tenuifolia leaves under different light intensities were analyzed, identifying 57 candidate transcription factors that influence monoterpene synthesis. Among these, 7 members of the StWRKY gene family were identified and mapped onto chromosomes using bioinformatics methods. The physicochemical properties of the proteins encoded by these StWRKY genes, their gene structures, and cis-acting elements were also studied. Comparative genomics and phylogenetic analyses revealed that Sch000013479 is closely related to AaWRKY1, AtWRKY41, and AtWRKY53, and it was designated as StWRKY1. Upon silencing and overexpressing the StWRKY1 transcription factor in S. tenuifolia leaves, changes in the expression of key genes in the menthone-type monoterpene synthesis pathway were observed. Specifically, when StWRKY1 was effectively silenced, the content of (-)-pulegone significantly decreased. These results enhance our understanding of the impact of StWRKYs on monoterpene synthesis in S. tenuifolia and lay the groundwork for further exploration of the regulatory mechanisms involved in the biosynthesis of menthone-type monoterpenes.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Luz , Monoterpenos , Proteínas de Plantas , Factores de Transcripción , Monoterpenos/metabolismo , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Lamiaceae/genética , Lamiaceae/metabolismo , Filogenia , Hojas de la Planta/metabolismo , Hojas de la Planta/genéticaRESUMEN
Biosynthetic gene clusters (BGCs) are regions of a genome where genes involved in a biosynthetic pathway are in proximity. The origin and evolution of plant BGCs as well as their role in specialized metabolism remain largely unclear. In this study, we have assembled a chromosome-scale genome of Japanese catnip (Schizonepeta tenuifolia) and discovered a BGC that contains multiple copies of genes involved in four adjacent steps in the biosynthesis of p-menthane monoterpenoids. This BGC has an unprecedented bipartite structure, with mirrored biosynthetic regions separated by 260 kilobases. This bipartite BGC includes identical copies of a gene encoding an old yellow enzyme, a type of flavin-dependent reductase. In vitro assays and virus-induced gene silencing revealed that this gene encodes the missing isopiperitenone reductase. This enzyme evolved from a completely different enzyme family to isopiperitenone reductase from closely related Mentha spp., indicating convergent evolution of this pathway step. Phylogenomic analysis revealed that this bipartite BGC has emerged uniquely in the S. tenuifolia lineage and through insertion of pathway genes into a region rich in monoterpene synthases. The cluster gained its bipartite structure via an inverted duplication. The discovered bipartite BGC for p-menthane biosynthesis in S. tenuifolia has similarities to the recently described duplicated p-menthane biosynthesis gene pairs in the Mentha longifolia genome, providing an example of the convergent evolution of gene order. This work expands our understanding of plant BGCs with respect to both form and evolution, and highlights the power of BGCs for gene discovery in plant biosynthetic pathways.
Asunto(s)
Lamiaceae , Familia de Multigenes , Monoterpenos , CromosomasRESUMEN
BACKGROUND: Venetoclax monotherapy is an effective option for patients with acute myeloid leukemia (AML). Venetoclax has also been used in non-myeloablative conditioning allogeneic hematopoietic stem cell transplantation (allo-HSCT) for high-risk AML with a tolerable toxicity profile. However, the efficacy and safety of a venetoclax-containing myeloablative conditioning (MAC) allo-HSCT regimen for high-risk AML have not been evaluated. OBJECTIVE: To evaluate the safety and efficacy of a MAC regimen containing venetoclax for high-risk AML. STUDY DESIGN: From 25 February 2021 to 4 September 2022, a total of 31 patients with high-risk AML who underwent allo-HSCT and a MAC regimen with venetoclax were analyzed. RESULTS: At the time of transplantation, 21 patients were in first complete remission (CR1), 4 were in a second complete remission (CR2), and 6 in non-remission (NR). Twenty-four patients (77.4%) were minimal residual disease (MRD)-positive before transplant. The FLT3-ITD gene mutation was present in 51.6% of patients. NUP98 rearrangement, MLL rearrangement or MLL-PTD and DEK::CAN fusion genes were found in 5 (16.1%), 7(22.6%) and 2 (6.5%) patients, respectively. Twenty-nine (93.6%) patients underwent haploidentical allo-HSCT. The median follow-up time was 278 days (range: 52-632 days). The 100-day cumulative incidence of grade 3 to 4 acute graft-versus-host disease (aGVHD) was 16.1% (95%CI, 7.2-36.0%). The 180-day cumulative incidence of moderate to severe chronic graft-versus-host disease (cGVHD) was 7.1% (95%CI, 1.9-26.9%). Cumulative incidence of 100-day cytomegalovirus (CMV) viraemia and 100-day Epstein-Barr virus (EBV) viraemia was 61.6% (95%CI, 46.5-81.4%) and 3.2% (95%CI, 0.4-22.2%), respectively. The 600-day overall survival (OS) and leukemia-free survival (LFS) were 80.9% (95%CI, 63.5-93.6%) and 81.3% (95%CI, 64.2-93.7%), respectively. The 600-day relapse incidence (RI) and non-relapse mortality (NRM) was 6.9% (95%CI, 1.8-26.3%) and 11.7% (95%CI, 3.9-35.0%). CONCLUSION: Our study shows that the addition of venetoclax to a MAC allo-HSCT was feasible, safe and effective for high-risk AML patients.
Asunto(s)
Infecciones por Citomegalovirus , Infecciones por Virus de Epstein-Barr , Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Leucemia Mieloide Aguda , Humanos , Infecciones por Virus de Epstein-Barr/complicaciones , Viremia/complicaciones , Estudios Retrospectivos , Trasplante Homólogo/efectos adversos , Herpesvirus Humano 4 , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/terapia , Enfermedad Injerto contra Huésped/epidemiología , Enfermedad Injerto contra Huésped/etiología , Enfermedad Injerto contra Huésped/prevención & control , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Acondicionamiento Pretrasplante/efectos adversos , Proteínas de Unión a Poli-ADP-Ribosa , Proteínas Cromosómicas no Histona , Proteínas OncogénicasRESUMEN
Purpose: Several Escherichia coli pathotypes still constitute an important public health concern owing to its pathogenicity and antimicrobial resistance. Moreover, biofilm formation of E. coli can allow the strains to interfere with host and antimicrobial eradication, thus conferring additional resistance. The association between the formation of biofilm and antimicrobial resistance determinants has been extensively exploited; nevertheless, there is still no definite conclusion. The purpose of this study was to provide additional data to augment the present knowledge about the subject. Methods: Antibiotic resistance/susceptibility profiles of 81 isolates from pediatric individuals in China between 2011 and 2014 against 20 antibiotics were assessed using the VITEK 2 system. Biofilm-forming capacities were evaluated using the crystal violet staining method, confocal laser scanning microscopy (CLSM), and field emission scanning electron microscopy. Biofilm compositions inside the biofilm formed by representative strains were assessed using CLSM. The effects of antibiotics on biofilms generated by E. coli strains of different biofilm-forming ability were examined using CLSM in combination with gatifloxacin. The relationships between antibiotic resistance, biofilm formation, and biofilm-specific resistance in E. coli isolates were investigated. Results: The results showed that 23 isolates were classified as multidrug-resistant, and 57 isolates were classified as extensively drug-resistant (XDR). Among the 69 isolates with the ability to form biofilms, 46 isolates were stronger biofilm formers. Correlation analysis demonstrated that strain populations exhibiting more robust biofilm formation likely contained larger proportions of XDR isolates. Conclusion: Together, our study implies that there was an association between biofilm-formation and resistance to several antibiotics for XDR-E. coli isolates, and would provide novel insights regarding the prevention and treatment against E. coli-related infections.