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1.
Sci Rep ; 9(1): 12673, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31481699

RESUMO

MADS-box transcription factors possess many functions in plant reproduction and development. However, few MADS-box genes related to secondary metabolites regulation have been identified. In Hevea brasiliensis, natural rubber is a representative cis-polyisoprenoids in secondary metabolism which occurs in the rubber laticifer cells, the molecular regulation basis of natural rubber biosynthesis is not clear. Here, a total of 24 MADS-box genes including 4 type I MADS-box genes and 20 type II MADS-box genes were identified in the transcriptome of rubber tree latex. The phylogenetic analysis was performed to clarify the evolutionary relationships of all the 24 rubber tree MADS-box proteins with MADS-box transcription factors from Arabidopsis thaliana and Oryza sativa. Four type I MADS-box genes were subdivided into Mα (3 genes) and Mß (1 gene). Twenty type II MADS-box genes were subclassified into MIKC* (8 genes) and MIKCc (12 genes). Eight MADS-box genes (HblMADS3, 5, 6, 7, 9, 13, 23, 24) were predominant expression in laticifers. ABA up-regulated the expression of HblMADS9, and the expression of HblMADS3, HblMADS5, HblMADS24 were up-regulated by MeJA. The function of HblMADS24 was elucidated. HblMADS24 bound HbFPS1 promoter in yeast and HblMADS24 activated HbFPS1 promoter in tobacco plants. Moreover, we proposed that HblMADS24 is a transcription activator of HbFPS1 which taking part in natural rubber biosynthesis.

2.
Genet Mol Biol ; 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31441928

RESUMO

Histone methylation plays a crucial role in various biological processes including from heterochromatin formation to transcriptional regulation. Currently, no information is available regarding histone methylation modifiers in the important rubber-producing plant Hevea brasiliensis. Here, we identified 47 histone methyltransferase (HMT) genes and 25 histone demethylase (HDM) genes, as possible members of the histone methylation modifiers in the rubber tree genome. According to the structural features of HMT and HDM, the HbHMTs were classified to two groups (HbPRMs and HbSDGs), the HbHDMs have two groups (HbLSDs and HbJMJs). The expression patterns of HbHMTs and HbHDMs were analyzed in five different tissues and at different phases of somatic embryogenesis. HbSDG10, 21, 25, 33, HbJMJ2, 18, 20 were with high expression at different phases of somatic embryogenesis. HbSDG10,14, 20, 21, 3 and HbPRMT4 were expressed highly in anther, HbSDG14, 20, 21, 22, 23, 33, 35 and HbPRMT1 HbJMJ7 and HbLSD1, 2, 3, 4 showed a high expression level in callus. HbSDG1, 7, 10, 13, 14, 18, 19, 21, 22, 23, 35, HbPRMT1, 8, HbJMJ5, 7, 11, 16, 20 and HbLSD2, 3, 4 were expressed highly in somatic embryo. HbSDG10, 21, 25, 33, HbLSD2, 3 3 were expressed highly in bud of regenerated plant. The expression analyses reveal that HbHMTs and HbHDMs exhibit different expression patterns at different phases during somatic embryogenesis and imply that some HbHMTs and HbHDMs play important roles during somatic embryogenesis. This study will provide fundamental to further study on regulations of histone metylation in Hevea brasiliensis.

3.
J Exp Bot ; 69(8): 1903-1912, 2018 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-29432591

RESUMO

Hevea brasiliensis is a key commercial source of natural rubber (cis 1,4-polyisoprene). In H. brasiliensis, rubber transferase is responsible for cis-1,4-polymerization of isoprene units from isopentenyl diphosphate and thus affects the yield of rubber. Little is known about the regulatory mechanisms of the rubber transferase gene at a molecular level. In this study we show that the 5'UTR intron of the promoter of the rubber transferase gene (HRT2) suppresses the expression of HRT2. A H. brasiliensis RING zinc finger protein (designated as HbRZFP1) was able to interact specifically with the HRT2 promoter to down-regulate its transcription in vivo. A 14-3-3 protein (named as HbGF14a) was identified as interacting with HbRZFP1, both in yeast and in planta. Transient co-expression of HbGF14a and HbRZFP1-encoding cDNAs resulted in HbRZFP1-mediated HRT2 transcription inhibition being relieved. HbGF14a repressed the protein-DNA binding of HbRZFP1 with the HRT2 promoter in yeast. We propose a regulatory mechanism by which the binding of HbGF14a to HbRZFP1 interferes with the interaction of HbRZFP1 with the HRT2 promoter, thereby repressing the protein-DNA binding between them. This study provides new insights into the role of HbGF14a in mediating expression of the rubber transferase gene in Hevea brasiliensis.


Assuntos
Proteínas 14-3-3/metabolismo , Regulação Enzimológica da Expressão Gênica , Hevea/metabolismo , Proteínas de Plantas/metabolismo , Transferases/genética , Proteínas 14-3-3/química , Proteínas 14-3-3/genética , Sequência de Aminoácidos , Regulação da Expressão Gênica de Plantas , Hevea/química , Hevea/classificação , Hevea/genética , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Regiões Promotoras Genéticas , Ligação Proteica , Domínios RING Finger , Borracha/metabolismo , Alinhamento de Sequência , Transferases/química , Transferases/metabolismo , Dedos de Zinco
4.
J Plant Res ; 131(3): 555-562, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29234988

RESUMO

Dracaena cambodiana is a traditional medicinal plant used for producing dragon's blood. The plants and dragon's blood of D. cambodiana contain a rich variety of steroidal saponins. However, little is known about steroidal saponin biosynthesis and its regulation in D. cambodiana. Here, 122 genes encoding enzymes involved in steroidal saponin biosynthesis were identified based on transcriptome data, with 29 of them containing complete open reading frames (ORF). Transcript expression analysis revealed that several genes related to steroidal saponin biosynthesis showed distinct tissue-specific expression patterns; the expression levels of genes encoding the key enzymes involved in the biosynthesis and early modification of steroidal saponins were significantly down-regulated in the stems in response to the inducer of dragon's blood, exhibiting positive correlations with the content of steroidal saponins. These results provide insights on the steroidal saponins biosynthetic pathway and mechanisms underlying induced formation of dragon's blood in D. cambodiana.


Assuntos
Dracaena/genética , Saponinas/biossíntese , Transcriptoma , Vias Biossintéticas , Dracaena/química , Dracaena/metabolismo , Perfilação da Expressão Gênica , Anotação de Sequência Molecular , Extratos Vegetais/biossíntese , Extratos Vegetais/química , Saponinas/química
5.
Front Plant Sci ; 8: 1974, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29187861

RESUMO

MYB transcription factors hold vital roles in the regulation of plant secondary metabolic pathways. Laticifers in rubber trees (Hevea brasiliensis) are of primary importance in natural rubber production because natural rubber is formed and stored within these structures. To understand the role of MYB transcription factors in the specialized cells, we identified 44 MYB genes (named HblMYB1 to HblMYB44) by using our previously obtained transcriptome database of rubber tree laticifer cells and the public rubber tree genome database. Expression profiles showed that five MYB genes were highly expressed in the laticifers. HblMYB19 and HblMYB44 were selected for further study. HblMYB19 and HblMYB44 bound the promoters of HbFDPS1, HbSRPP, and HRT1 in yeast. Furthermore, the transient overexpression of HblMYB19 and HblMYB44 in tobacco plants significantly increased the activity of the promoters of HbFDPS1, HbSRPP, and HRT1. Basing on this information, we proposed that HblMYB19 and HblMYB44 are the regulators of HbFDPS1, HbSRPP, and HRT1, which are involved in the biosynthesis pathway of natural rubber.

6.
Sci Rep ; 7: 45157, 2017 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-28332623

RESUMO

Abscisic acid (ABA) is an essential phytohormone involved in diverse physiological processes. Although genome-wide analyses of the ABA receptor PYR/PYL/RCAR (PYL) protein/gene family have been performed in certain plant species, little is known about the ABA receptor protein/gene family in the rubber tree (Hevea brasiliensis). In this study, we identified 14 ABA receptor PYL proteins/genes (designated HbPYL1 through HbPYL14) in the most recent rubber tree genome. A phylogenetic tree was constructed, which demonstrated that HbPYLs can be divided into three subfamilies that correlate well with the corresponding Arabidopsis subfamilies. Eight HbPYLs are highly expressed in laticifers. Five of the eight genes are simultaneously regulated by ABA, jasmonic acid (JA) and ethylene (ET). The identification and characterization of HbPYLs should enable us to further understand the role of ABA signal in the rubber tree.


Assuntos
Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Hevea/genética , Hevea/metabolismo , Família Multigênica , Reguladores de Crescimento de Planta/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Sequência Conservada , Evolução Molecular , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Estudo de Associação Genômica Ampla , Hevea/classificação , Filogenia , Reguladores de Crescimento de Planta/farmacologia , Regiões Promotoras Genéticas , Borracha/metabolismo , Transcriptoma
7.
Sci Rep ; 6: 38315, 2016 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-27922066

RESUMO

Dragon's blood is a red resin mainly extracted from Dracaena plants, and has been widely used as a traditional medicine in East and Southeast Asia. The major components of dragon's blood are flavonoids. Owing to a lack of Dracaena plants genomic information, the flavonoids biosynthesis and regulation in Dracaena plants remain unknown. In this study, three cDNA libraries were constructed from the stems of D. cambodiana after injecting the inducer. Approximately 266.57 million raw sequencing reads were de novo assembled into 198,204 unigenes, of which 34,873 unique sequences were annotated in public protein databases. Many candidate genes involved in flavonoid accumulation were identified. Differential expression analysis identified 20 genes involved in flavonoid biosynthesis, 27 unigenes involved in flavonoid modification and 68 genes involved in flavonoid transport that were up-regulated in the stems of D. cambodiana after injecting the inducer, consistent with the accumulation of flavonoids. Furthermore, we have revealed the differential expression of transcripts encoding for transcription factors (MYB, bHLH and WD40) involved in flavonoid metabolism. These de novo transcriptome data sets provide insights on pathways and molecular regulation of flavonoid biosynthesis and transport, and improve our understanding of molecular mechanisms of dragon's blood formation in D. cambodiana.


Assuntos
Dracaena/genética , Flavonoides/biossíntese , Regulação da Expressão Gênica de Plantas , Extratos Vegetais/química , Proteínas de Plantas/genética , Transcriptoma , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Dracaena/metabolismo , Flavonoides/genética , Biblioteca Gênica , Ontologia Genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Medicina Tradicional , Anotação de Sequência Molecular , Extratos Vegetais/biossíntese , Extratos Vegetais/genética , Extratos Vegetais/isolamento & purificação , Proteínas de Plantas/metabolismo , Caules de Planta/genética , Caules de Planta/metabolismo , Repetições WD40
8.
Front Plant Sci ; 7: 1709, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27895659

RESUMO

In plants MADS-box transcription factors (TFs) play important roles in growth and development. However, no plant MADS-box gene has been identified to have a function related to secondary metabolites regulation. Here, a MADS-box TF gene, designated as HbMADS4, was isolated from Hevea brasiliensis by the yeast one-hybrid experiment to screen the latex cDNA library using the promoter of the gene encoding H. brasiliensis small rubber particle protein (HbSRPP) as bait. HbMADS4 was 984-bp containing 633-bp open reading frame encoding a deduced protein of 230 amino acid residues with a typical conserved MADS-box motif at the N terminus. HbMADS4 was preferentially expressed in the latex, but little expression was detected in the leaves, flowers, and roots. Its expression was inducible by methyl jasmonate and ethylene. Furthermore, transient over-expression and over-expression of HbMADS4 in transgenic tobacco plants significantly suppressed the activity of the HbSRP promoter. Altogether, it is proposed that HbMADS4 is a negative regulator of HbSRPP which participates in the biosynthesis of natural rubber.

9.
Front Plant Sci ; 7: 1204, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27555864

RESUMO

Rubber tree (Hevea brasiliensis) self-rooting juvenile clones (JCs) are promising planting materials for rubber production. In a comparative trial between self-rooting JCs and donor clones (DCs), self-rooting JCs exhibited better performance in rubber yield. To study the molecular mechanism associated with higher rubber yield in self-rooting JCs, we sequenced and comparatively analyzed the latex of rubber tree self-rooting JCs and DCs at the transcriptome level. Total raw reads of 34,632,012 and 35,913,020 bp were obtained from the library of self-rooting JCs and DCs, respectively, by using Illumina HiSeq 2000 sequencing technology. De novo assemblies yielded 54689 unigenes from the library of self-rooting JCs and DCs. Among 54689 genes, 1716 genes were identified as differentially expressed between self-rooting JCs and DCs via comparative transcript profiling. Functional analysis showed that the genes related to the mass of categories were differentially enriched between the two clones. Several genes involved in carbohydrate metabolism, hormone metabolism and reactive oxygen species scavenging were up-regulated in self-rooting JCs, suggesting that the self-rooting JCs provide sufficient molecular basis for the increased rubber yielding, especially in the aspects of improved latex metabolisms and latex flow. Some genes encoding epigenetic modification enzymes were also differentially expressed between self-rooting JCs and DCs. Epigenetic modifications may lead to gene differential expression between self-rooting JCs and DCs. These data will provide new cues to understand the molecular mechanism underlying the improved rubber yield of H. brasiliensis self-rooting clones.

10.
Plant Physiol Biochem ; 104: 304-11, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27208821

RESUMO

Dragon's blood is a traditional medicine widely used in the world, and the main components of which are flavonoids. However, little is known about its formation mechanism. Previous studies indicate that plant glutathione S-transferase (GST) genes are involved in transportation of flavonoids from cytosolic synthesis to vacuolar accumulation. In this study, 20 Dracaena cambodiana GST genes (DcGSTs) were identified based on transcriptome database. Phylogenetic analysis revealed that 20 DcGSTs belonged to seven different classes. Tissue-specific expression analysis suggested that DcGSTs displayed differential expressions either in their transcript abundance or expression patterns under normal growth conditions. The transcript profiles of three DcGSTs in response to the inducer of dragon's blood were strongly correlated with flavonoids biosynthetic genes, consistent with dragon's blood accumulation. Our survey provides useful information for future studies on GST genes involved in dragon's blood formation in D. cambodiana.


Assuntos
Dracaena/enzimologia , Dracaena/genética , Flavonoides/metabolismo , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Glutationa Transferase/genética , Transcriptoma/genética , Flavonoides/biossíntese , Perfilação da Expressão Gênica , Glutationa Transferase/metabolismo , Especificidade de Órgãos/genética , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
11.
Int J Mol Sci ; 16(2): 4136-49, 2015 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-25690030

RESUMO

Ethylene is an important factor that stimulates Hevea brasiliensis to produce natural rubber. 1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is a rate-limiting enzyme in ethylene biosynthesis. However, knowledge of the ACS gene family of H. brasiliensis is limited. In this study, nine ACS-like genes were identified in H. brasiliensis. Sequence and phylogenetic analysis results confirmed that seven isozymes (HbACS1-7) of these nine ACS-like genes were similar to ACS isozymes with ACS activity in other plants. Expression analysis results showed that seven ACS genes were differentially expressed in roots, barks, flowers, and leaves of H. brasiliensis. However, no or low ACS gene expression was detected in the latex of H. brasiliensis. Moreover, seven genes were differentially up-regulated by ethylene treatment. These results provided relevant information to help determine the functions of the ACS gene in H. brasiliensis, particularly the functions in regulating ethylene stimulation of latex production.


Assuntos
Hevea/genética , Liases/genética , Sequência de Aminoácidos , Clonagem Molecular , Etilenos/farmacologia , Genes de Plantas , Hevea/enzimologia , Liases/classificação , Liases/metabolismo , Dados de Sequência Molecular , Filogenia , Regiões Promotoras Genéticas , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Alinhamento de Sequência , Regulação para Cima/efeitos dos fármacos
12.
Zhongguo Zhong Yao Za Zhi ; 39(7): 1260-4, 2014 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-25011265

RESUMO

The latest findings of our laboratory showed that Angelica sinensis polysaccharide (ASP) showed a definite effect in regulating the aging of hematopoietic stem cells. Leukemia is a type of malignant hematopoietic tumor in hematopoietic stem cells. There have been no relevant reports about ASP's effect in regulating the aging of leukemia cells. In this study, human acute myeloid leukemia (AML) KG1alpha cell lines in logarithmic growth phase were taken as the study object, and were divided into the ASP group, the cytarabine (Ara-C) group, the ASP + Ara-C group and the control group. The groups were respectively treated with different concentration of ASP, Ara-C and ASP + Ara-C for different periods, with the aim to study the effect of ASP combined with Ara-C in regulating the aging of human acute myeloid leukemia KG1alpha cell lines and its relevant mechanism. The results showed that ASP, Ara-C and ASP + Ara-C could obviously inhibit KG1alpha cell proliferation in vitro, block the cells in G0/G1 phase. The cells showed the aging morphological feature. The percentage of positive stained aging cells was dramatically increased, and could significantly up-regulate the expression of aging-related proteins P16 and RB, which were more obvious in the ASP + Ara-C group. In conclusion, the aging mechanism of KG1alpha cell induced by ASP and Ara-C may be related to the regulation of the expression of aging-related proteins, suggesting that the combined administration of ASP and anticancer drugs plays a better role in the treatment of leukemia .


Assuntos
Envelhecimento/efeitos dos fármacos , Angelica sinensis/química , Leucemia/fisiopatologia , Polissacarídeos/farmacologia , Envelhecimento/genética , Envelhecimento/metabolismo , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Inibidor p16 de Quinase Dependente de Ciclina/genética , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Humanos , Leucemia/tratamento farmacológico , Leucemia/genética , Leucemia/metabolismo , Proteína do Retinoblastoma/genética , Proteína do Retinoblastoma/metabolismo , Células Tumorais Cultivadas
13.
Int J Cancer ; 135(11): 2604-11, 2014 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-24740547

RESUMO

Baseline human papillomavirus (HPV) prevalence and type distribution were evaluated in young Chinese women enrolled in a clinical trial of an HPV vaccine (ClinicalTrials.gov registration NCT00779766). Cervical specimens and blood samples were collected at baseline from women aged 18-25 years (n = 6,051) from four sites across Jiangsu province. Cervical specimens were tested for HPV DNA by SPF10 PCR-DEIA-LiPA25 version 1, and HPV-16/18 type-specific polymerase chain reaction. Anti-HPV-16 and anti-HPV-18 antibody titres were quantified by enzyme-linked immunosorbent assay. At baseline, 15.3% of women were DNA positive for any of 14 HPV high-risk (hr) types (HPV-16/18/31/33/35/39/45/51/52/56/58/59/66/68). The most commonly detected hrHPV types in cervical specimens were HPV-52 (4.0%) and HPV-16 (3.7%). High-risk HPV DNA-positivity increased with severity of cytological abnormalities: 39.3% in atypical squamous cells of undetermined significance, 85.0% in low-grade squamous intraepithelial lesions and 97.8% in high-grade squamous intraepithelial lesions (HSIL). The hrHPV types most frequently detected in HSIL were HPV-16 (63.0%), HPV-18 (17.4%), HPV-52 (17.4%), HPV-58 (15.2%) and HPV-33 (15.2%). The hrHPV types most frequently detected in cervical intraepithelial neoplasia 2+ were HPV-16 (66.1%), HPV-33 (16.1%), HPV-52 (16.1%), HPV-58 (14.5%) and HPV-51 (11.3%). Multiple hrHPV infections were reported for 24.4% of hrHPV DNA positive women. Regardless of baseline HPV DNA status, 30.5% and 16.0% of subjects were initially seropositive for anti-HPV-16 and anti-HPV-18, respectively. In conclusion, the high baseline seropositivity rate and intermediate prevalence of cervical hrHPV types in Chinese women aged 18-25 years underlines the importance of early HPV vaccination in this population.


Assuntos
Neoplasia Intraepitelial Cervical/epidemiologia , Papillomaviridae/classificação , Infecções por Papillomavirus/epidemiologia , Displasia do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/epidemiologia , Saúde da Mulher , Adolescente , Adulto , Neoplasia Intraepitelial Cervical/prevenção & controle , Neoplasia Intraepitelial Cervical/virologia , Colo do Útero/patologia , China/epidemiologia , Ensaios Clínicos Fase II como Assunto , Ensaios Clínicos Fase III como Assunto , DNA Viral/genética , Método Duplo-Cego , Ensaio de Imunoadsorção Enzimática , Feminino , Seguimentos , Humanos , Estadiamento de Neoplasias , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/prevenção & controle , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/uso terapêutico , Reação em Cadeia da Polimerase , Prevalência , Prognóstico , Estudos Prospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto , Displasia do Colo do Útero/prevenção & controle , Displasia do Colo do Útero/virologia , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/virologia , Esfregaço Vaginal , Adulto Jovem
14.
Zhongguo Zhong Yao Za Zhi ; 39(1): 121-5, 2014 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-24754180

RESUMO

Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.


Assuntos
Angelica/química , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Leucemia/tratamento farmacológico , Fígado/efeitos dos fármacos , Animais , Linhagem Celular Tumoral , Citarabina/administração & dosagem , Humanos , Células K562 , Masculino , Camundongos , Camundongos SCID , Polissacarídeos/administração & dosagem
15.
Zhongguo Zhong Yao Za Zhi ; 39(22): 4442-7, 2014 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-25850282

RESUMO

Neurodegenerative disease is common and frequently occurs in elderly patients. Previous studies have shown that ginsenoside Rg1 was able to inhibit senescent of brain, but the mechanism on the brain during the treatment remains elucidated. To study the mechanism of ginsenoside Rg1 in the process of anti-aging of brain, forty male SD rats were randomly divided into normal group, Rg1 normal group, brain aging model group and Rg1 brain aging model group, each group with 10 rats (brain aging model group: subcutaneous injection of D-galactose (120 mg kg(-1)), qd for 42 consecutive days; Rg1 brain aging model group: while copying the same test as that of brain aging model group, begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Rg1 normal group: subcutaneous injection of the same amount of saline; begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Normal: injected with an equal volume of saline within the same time. Perform the related experiment on the second day after finishing copying the model or the completion of the first two days of drug injections). Learning and memory abilities were measured by Morris water maze. The number of senescent cells was detected by SA-beta-Gal staining while the level of IL-1 and IL-6 proinflammatory cytokines in hippocampus were detected by ELISA. The activities of SOD, contents of GSH in hippo- campus were quantified by chromatometry. The change of telomerase activities and telomerase length were performed by TRAP-PCR and southern blotting assay, respectively. It is pointed that, in brain aging model group, the spatial learning and memory capacities were weaken, SA-beta-Gal positive granules increased in section of brain tissue, the activity of antioxidant enzyme SOD and the contents of GSH decreased in hippocampus, the level of IL-1 and IL-6 increased in hippocampus, while the length of telomere and the activity of telomerase decreased in hippocampus. Rats of Rg1 brain aging group had their spatial learning and memory capacities enhanced, SA-beta-Gal positive granules in section of brain tissue decreased, the activity of antioxidant enzyme SOD and the contents of GSH increased in hippocampus, the level of IL-1 and IL-6 in hippocampus decreased, the length contraction of telomere suppressed while the change of telomerase activity increased in hippocampus. Compared with that of normal group, the spatial learning and memory capacities were enhanced in Rg1 normal group, SA-beta-Gal positive granules in section of brain tissue decreased in Rg1 normal group, the level of IL-1 and IL-6 in hippocampus decreased in Rg1 normal group. The results indicated that improvement of antioxidant ability, regulating the level of proinflammatory cytokines and regulation of telomerase system may be the underlying anti-aging mechanism of Ginsenoside Rg1.


Assuntos
Envelhecimento/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Ginsenosídeos/farmacologia , Animais , Masculino , Ratos , Ratos Sprague-Dawley
16.
Biosci Biotechnol Biochem ; 74(11): 2183-8, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21071842

RESUMO

Calcium-dependent protein kinases (CDPKs), as major primary Ca(2+) sensors, have been implicated in the regulation of stress and developmental signals in plants. In this study, a novel CDPK gene, designated HbCDPK1, was isolated from Hevea brasiliensis. The HbCDPK1 cDNA had 2,400 bp with an open reading frame of 1,671 bp encoding 556 amino acids, and the deduced HbCDPK1 protein contained four characteristic domains identified in CDPKs, showing a high level of sequence similarity to CDPKs from other plants. Expression analysis revealed more significant accumulation of the transcripts of HbCDPK1 in latex than in the leaves, bark, and roots in H. brasiliensis. In addition, transcription of HbCDPK1 was strongly induced by mechanical wounding, jasmonic acid (JA), and ethephon. Recombinant HbCDPK1 was expressed in E. coli, and its activity was assayed. The assay indicated that HbCDPK1 had the kinase and Ca(2+)-binding activity in vitro as a calcium-dependent protein. The potential roles of the HbCDPK1 are discussed as to latex production and rubber biosynthesis.


Assuntos
Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Hevea/enzimologia , Compostos Organofosforados/farmacologia , Proteínas Quinases/genética , DNA Complementar/isolamento & purificação , Perfilação da Expressão Gênica , Genes de Plantas , Hevea/genética , Látex/biossíntese , Fases de Leitura Aberta/genética , Reguladores de Crescimento de Planta , Estruturas Vegetais/química , RNA Mensageiro/análise , Borracha
17.
Zhonghua Liu Xing Bing Xue Za Zhi ; 30(6): 611-5, 2009 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-19957630

RESUMO

OBJECTIVE: Feasibility of using MNA cell-culture inoculation test to detect and isolate the street rabies virus. METHODS: Using MNA cell-culture inoculation test, fluorescent antibody test (FAT) and sandwich ELISA with double-antibodies to detect 33 specimens of street rabies virus, 20 specimens of negative canine brains and 4 specimens of healthy mice brains. RESULTS: 33 specimens of street rabies virus were positive to the cell-culture inoculation test but the others were negative. The concordances of MNA cell-cultured inoculation test with FAT and sandwich ELISA with double-antibodies were both 100%. CONCLUSION: MNA cell-culture inoculation test appeared to be both highly sensitive and specific in detecting the street rabies virus, and could be used in detection and isolation of the virus.


Assuntos
Técnicas de Cultura de Células/métodos , Vírus da Raiva/isolamento & purificação , Animais , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Estudos de Viabilidade , Imunofluorescência/métodos , Camundongos , Sensibilidade e Especificidade
18.
Plant Physiol Biochem ; 47(10): 954-9, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19577934

RESUMO

Hsp70s have been shown to play important roles in helping cells to cope with adverse environments, especially in response to temperature. In this study a novel ethephon-induced Hsp gene, designated as HbHsp70, was isolated from Hevea brasiliensis. The HbHsp70 cDNA contained a 1965 bp open reading frame encoding 655 amino acids. The deduced HbHsp70 protein showed high identities to Hsp70s from other plants. Expression studies revealed more significant accumulation of HbHsp70 transcripts in leaves and stems than in roots, barks and latex. The transcription of HbHsp70 was induced by ethephon, heat treatment and low temperature stress, whereas jasmonic acid had little effects. Recombinant HbHsp70 was expressed in Escherichia coli and purified by Ni-NTA affinity chromatography. Measuring the light scattering of luciferase (Luc) revealed that HbHsp70 prevents the aggregation of luc during high-temperature stress. In vitro experiments showed that HbHsp70 had protective functions not only against heat stress but also against chilling stress. All these data suggest that HbHsp70 may play roles in responses to heat shock and low temperature in H. brasiliensis.


Assuntos
Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/genética , Hevea/genética , Compostos Organofosforados/farmacologia , Proteínas de Plantas/genética , Sequência de Aminoácidos , Clonagem Molecular , Temperatura Baixa , DNA Complementar/química , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Perfilação da Expressão Gênica , Proteínas de Choque Térmico HSP70/metabolismo , Hevea/metabolismo , Temperatura Alta , Chaperonas Moleculares/metabolismo , Dados de Sequência Molecular , Reguladores de Crescimento de Planta/farmacologia , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
19.
Artigo em Chinês | MEDLINE | ID: mdl-19105347

RESUMO

OBJECTIVE: To understand the evolution of HIV-1 CRF07_BC envelope, we performed a longitudinal study on two patients during their early HIV-1 infection. METHODS: RNA was extracted from the plasma of the individuals and the C2-C5 fragments of the gp120 gene of HIV-1 were amplified by RT-PCR. Purified DNA segments were inserted into T easy vector and transformed into E. coli Top 10 competent cells. Positive clones were identified by blue-white screening, confirmed by PCR and sequenced by ABI 3700. RESULTS: The samples were collected from the patients every 6 months from seroconversion time. The genetic diversity and divergence in env gene showed consistent increases over time. Our sequence analysis also revealed obvious non-synonymous change in env C1, C3 and V4 regions among these samples. CONCLUSION: The results support the concept that the consistent pattern of viral evolution existed during early phase of HIV-1 infection. C1, C3 and V4 region of env gene may be mainly immunological target during AIDS progression.


Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Adulto , HIV-1/isolamento & purificação , Humanos , Estudos Longitudinais , Masculino
20.
J Biosci ; 33(5): 681-90, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19179756

RESUMO

The full-length cDNA encoding a cysteine protease,designated HbCP1, was isolated for the first time from Hevea brasiliensis by the rapid amplification of cDNA ends (RACE) method. HbCP1 contained a 1371 bp open reading frame encoding 457 amino acids.The deduced HbCP1 protein,which showed high identity to cysteine proteases of other plant species,was predicted to possess a putative repeat in toxin (RTX) domain at the N-terminal and a granulin (GRAN) domain at the C-terminal.Southern blot analysis indicated that the HbCP1 gene is present as a single copy in the rubber tree.Transcription pattern analysis revealed that HbCP1 had high transcription in laticifer,and low transcription in bark and leaf.The transcription of HbCP1 in latex was induced by ethylene and tapping.Cloning of the HbCP1 gene will enable us to further understand the molecular characterization of cysteine protease and its possible function in the rubber tree.


Assuntos
Clonagem Molecular , Cisteína Endopeptidases/genética , Hevea/enzimologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Cisteína Endopeptidases/metabolismo , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Hevea/genética , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/química
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