RESUMO
In this work, we report the syntheses of three Pt(II) emitters, namely, Pt4N1, Pt4N2, and Pt4N3, to which their tetradentate chelates were assembled by linking two pyrazolate chelates with a single xylenylamino entity. Functionalization of Pt4N1 was achieved upon addition of electronegative CF3 substituent on pyridinyl groups and switching to more electron deficient pyrazinyl groups in giving Pt4N2 and Pt4N3, respectively. The vertical arranged xylenylamino entity has effectively suppressed the inter-molecular π-π stacking and Pt···Pt interaction, as shown by the single crystal X-ray structural analyses. Upon fabrication of OLED devices, Pt4N2 and Pt4N3 based devices delivered efficient cyan and green emission, with an EQEmax of 15.2% and 11.2%, respectively, affirming the successfulness of the tetradentate chelating strategy.
RESUMO
Artemisia stolonifera is a relative of A. argyi. The two species are difficult to be distinguished due to the similarity in leaf shape and have even less distinctive features after processing. This study aims to establish a method to quickly distinguish between them. At the same time, we examined the reasonability and applicability of the specific polymerase chain reaction(PCR) method. The C/T single nucleotide polymorphism was detected at the position 202 of the sequence, based on which specific primers were designed to identify these two species. The PCR with the specific primer JNC-F and the universal primer ITS3R produced a specific band at 218 bp for A. argyi and no band for A. stolonifera, which can be used to detect at least 3% of A. argyi samples mixed in A. stolonifera samples. The PCR with the specific primer KY-F and the universal primer ITS3R produced a specific band at 218 bp for A. stolonifera and no band for A. argyi, which can be used to detect at least 5% of A. stolonifera samples mixed with A. argyi. The limit of detection of the established method was 5 ng DNA. The established PCR method can accurately distinguish between A. stolonifera and A. argyi, which provides an experimental basis for the quality control of A. stolonifera and determines whether the herbs are adulterated.
Assuntos
Artemisia , Artemisia/genética , Tricomas , Reação em Cadeia da Polimerase , Técnicas de Amplificação de Ácido Nucleico , Folhas de Planta/genéticaRESUMO
Severe leaf spot on Polygonum cuspidatum was found in the planting base of P. cuspidatum in Fangxian county, Shiyan of Hubei province. To clarify the types of pathogens and their pathogenesis, the present study isolated and purified the pathogen of leaf spot disease of P. cuspidatum according to Koch's postulates, determined the pathogenicity of the pathogen, and investigated its biological characteristics. Meanwhile, the inhibitory effects of 11 types of fungicides on the bacteria were determined according to the mycelium growth rate, and suitable prevention and control drugs were selected. The results showed that the pathogen isolated from the diseased leaves of P. cuspidatum was Phoma rhei by morphological and molecular identification. The colony morphology and microscopic characteristics were the same as those of Ph. rhei. The homology of rDNA-ITS and TEF gene sequences with Ph. rhei reached 99.96% and 99.43%, respectively. The optimal growth temperature of Ph. rhei was 25 â, and the optimal pH was 7-10. Furthermore, Ph. rhei grew faster under dark or light conditions. In fungicides, 0.3% eugenol, 250 g·L~(-1) propiconazole, and 33.5% quinoline copper had significant inhibitory effects on the pathogen with EC_(50) values of 57.54, 59.58, 88.69 µg·mL~(-1), respectively. Eugenol is a botanical fungicide, which can be used as a green and environmentally friendly fungicide in the prevention and control of P. cuspidatum. This study reported for the first time that the pathogen of P. cuspidatum leaf spot was Ph. rhei. investigated the biological characteristics of the pathogen, and screened the indoor chemicals, which provided a theoretical basis for the prevention and control of P. cuspidatum leaf spot in production.
Assuntos
Fallopia japonica , Fungicidas Industriais , Ascomicetos , Eugenol , Fungicidas Industriais/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controleRESUMO
In this paper,the fingerprint of different varieties of chrysanthemum were established with " Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" and the content of chlorogenic acid,galuteolin and 3,5-O-dicaffeoylquinic acid in 29 batches of different varieties of chrysanthemum in Futianhe town,Huangtugang town and Wuhan city were compared. At the same time,similarity evaluation and common peak clustering analysis were carried out. There were 11 common peaks in the fingerprints of 29 batches of different varieties of chrysanthemum,and the similarity ranged from 0. 802 to 0. 975. Hangju and Gongju were divided into one group by cluster analysis,and Huangju into another category. The established fingerprint method provides a basis for the identification of chrysanthemum cultivars. The content of 29 batches of chlorogenic acid was between 4. 092 and 11. 723 mg·g-1,luteolin was between 1. 010 and 11. 713 mg·g-1,and 3,5-O-dicaffeoylquinic acid was between 8. 828 and 33. 435 mg·g-1,both reach the pharmacopoeia standard,but the effective components of different varieties of chrysanthemum were quite different. Based on the contents of three active ingredients and the diversity of fingerprint peaks,the quality of the characteristic germplasm resource of local Fubaijuin Macheng is superior,and the protection of local characteristic germplasm resource should be strengthened in production.
Assuntos
Chrysanthemum/química , Compostos Fitoquímicos/análise , Ácido Clorogênico/análogos & derivados , Ácido Clorogênico/análise , Cromatografia Líquida de Alta Pressão , Luteolina/análiseRESUMO
To study the effects of fresh-cut drying methods on the appearance and internal components of Panax notoginseng, and explore the feasibility of fresh-cut drying methods of P. notoginseng, so as to provide more effective processing methods for the production of P. notoginseng slices and Chinese patent medicines. In this study, we have compared the effects of 6 different drying methods on drying time, drying rate, density, appearance and internal components of P. notoginseng roots. It takes about 453 h to dry by whole-root drying in the sun, with a long constant speed period and a slow drying rate, the time of whole-root drying at 50 â was shortened by 61.6% compared with whole-root drying in the sun, which resulted in the decrease of density and poor appearance of the medicinal material with hollow and crack appeared in the xylem, while the drying time of fresh-cut drying method was reduced by 61.82% to 91.58% and the drying rate increased greatly, due to the relatively slow drying process in the sun or in the shade after fresh-cut, salting-out and whitening appeared on the surface, and the internal components were all decreased to some extent. The drying time of fresh-cut drying at 50 â was 91.58% and 68.83% shorter than that of whole-root drying in the sun and at 50 â, respectively. When drying at 50 â after fresh-cut, the appearance and content of internal components of the medicinal materials were better, the appearance was yellowish green, the cut sections were clear with uniform pore distribution, and the content of saponin components was increased by 7.24% compared with that of the whole-root drying at 50 â, When drying at 40 â, the surface of slices has salting-out and whitening spots, and the loss of dencichine and total sugar was large, but at 60 â, this high temperature made the rate of dehydration of slices was extremely fast, which led to severe cracking and fragmentation, and the loss of total sugar and alcohol extract was large. By vacuum freeze drying after fresh-cut, the structure of medicinal materials slices was loose, the density was greatly reduced, and the appearance was different from those recorded in traditional books. The contents of total saponin components and dencichine were increased by 16.51% and 22.54%, respectively, compared with traditional whole-root drying. The fresh-cut process method is feasible in the production of P. notoginseng slices. In production, it is recommended that drying at 50 â after fresh-cut can make the medicinal materials better in appearance and content of internal components, which is convenient for the subsequent processing and industrial feeding extraction. For the purpose of internal contents, it is better to adopt freeze-drying after fresh-cut processing method.
Assuntos
Dessecação , Medicamentos de Ervas Chinesas/normas , Panax notoginseng , Saponinas/análise , Liofilização , Raízes de Plantas , Controle de QualidadeRESUMO
Objective: To study the taxonomy and distribution of Chinese medicinal centipedes. Methods: The species of Chinese medicinal centipedes were investigated in the light of their morphology. According to the feature of life, the distribution of centipedes were explored. Results: There were 12 centipede species in China, and seven of them were used for medical, the species could be effectively distinguished by the identification key. It was suggested that their characteristics were related to the climatic factors such as temperature, humidity, altitude and air pressure. Conclusion: The distribution of medicinal centipede is characteristics of "three river system distribution belts" and "three geographical distribution areas". The results provide the basis for the development and application of medicinal centipedes in China.
Assuntos
Artrópodes , Altitude , Animais , China , Umidade , Plantas Medicinais , TemperaturaRESUMO
Kruppel-like factor 2 (KLF2) is a crucial anti-angiogenic factor. However, its precise role in hepatic angiogenesis induced by liver sinusoidal endothelial cells (LSECs) remain unclear. This study was aimed to evaluate the effect of KLF2 on angiogenesis of LSECs and to explore the corresponding mechanism. Cultured human LSECs were infected with different lentiviruses to overexpress or suppress KLF2 expression. The CCK-8 assay, transwell migration assay and tube formation test, were used to investigate the roles of KLF2 in the proliferation, migration and vessel tube formation of LSECs, respectively. The expression and phosphorylation of ERK1/2 were detected by western blot. We discovered that the up-regulation of KLF2 expression dramatically inhibited proliferation, migration and tube formation in treated LSECs. Correspondingly, down-regulation of KLF2 expression significantly promoted proliferation, migration and tube formation in treated LSECs. Additionally, KLF2 inhibited the phosphorylation of ERK1/2 pathway, followed by the function of KLF2 in the angiogenesis of LSECs disrupted. In conclusion, KLF2 suppressed the angiogenesis of LSECs through inhibition of cell proliferation, migration, and vessel tube formation. These functions of KLF2 may be mediated through the ERK1/2 signaling pathway.
Assuntos
Células Endoteliais/fisiologia , Fatores de Transcrição Kruppel-Like/metabolismo , Fígado/citologia , Fígado/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Neovascularização Fisiológica/fisiologia , Células Cultivadas , HumanosRESUMO
The aim of this paper is to apply Raman spectroscopy technique to develop rapid quantitative models for five kinds of Traditional Chinese Medicine containing CaCO3. In the experiment, Raman spectras of 67 batch of sample including Otolithum Sciaenae, Galaxeae Os, Ophicalcitum, Calcite, Stalactite and their mixture which had different content of CaCO3 were collected, and the quantitative models were established by using an improved siPLS to optimize the characteristic spectral bands and using the CaCO3 contents which were measured by EDTA titration method as references. Compared with the results by EDTA titration, the established quantitative model for CaCO, content showed a prediction result that the average relative deviation of the prediction results is 2. 71% and the average recovery rate was 100.46%, when the content is between 0.465 4-0.999 7, and when the characteristic spectral bands of 1 290-1 280, 730-714, 700-690, 660-650, 465-460, 455-445, 405-385 cm(-1) had been optimized. The result also showed that the model using Raman spectroscopy and based on an improved siPLS can get a rapid determination for contents of 5 kinds of Traditional Chinese Medicine containing CaCO3.
Assuntos
Carbonato de Cálcio/química , Medicamentos de Ervas Chinesas/química , Plantas Medicinais/química , Análise Espectral Raman/métodos , Análise dos Mínimos Quadrados , Modelos EstatísticosRESUMO
BACKGROUND: Human epidermal growth factor receptor 2 (HER2) is an important proto-oncogene of prognostic use in gastric cancer (GC). Fluorescence in-situ hybridization (FISH) and immunohistochemistry (IHC) are the main clinical methods of detection of HER2, but consistency between the methods is poor and the cause of the discrepancy is unclear. AIM: To investigate the involvement of HER2 mRNA status in the disparity between gene amplification and protein overexpression. METHODS: We investigated HER2 gene, mRNA, and protein profiles in gastric precancer and cancer tissues by use of the molecular approaches FISH, real-time polymerase chain reaction, and IHC. The relationships between HER2 and matrix metalloproteinase 9 (MMP9) and Smad7 expression were analyzed and the involvement of HER2 in the interaction between tumor cells and lymphocytes was investigated by coculturing GC cell lines with peripheral blood mononuclear cells (PBMCs). RESULTS: HER2 protein expression was significantly increased in cancer compared with precancer (P = 0.003), and the corresponding mRNA levels were significantly lower in precancer and cancer tissues than in normal tissues (κ = 0.290, P = 0.025). HER2 mRNA levels were significantly higher in tumor than in peritumor tissue (P = 0.028), and were positively correlated with MMP9 and Smad7 mRNA levels in tumor tissues. HER2 mRNA expression in GC cell lines was increased by coculture with PBMCs. CONCLUSIONS: Different HER2 mRNA profiles, possibly in relation to contact between tumor cells and lymphocytes, might help to explain the discrepancy between gene amplification and protein overexpression results.
Assuntos
Biomarcadores Tumorais/genética , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , RNA Mensageiro/metabolismo , Receptor ErbB-2/genética , Neoplasias Gástricas/genética , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Estudos de Casos e Controles , Linhagem Celular Tumoral , Técnicas de Cocultura , Feminino , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Leucócitos Mononucleares/metabolismo , Masculino , Metaloproteinase 9 da Matriz/genética , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Proto-Oncogene Mas , Reação em Cadeia da Polimerase em Tempo Real , Receptor ErbB-2/metabolismo , Proteína Smad7/genética , Neoplasias Gástricas/enzimologia , Regulação para CimaRESUMO
This paper reviewed the emergence process of the subject and methodology of Chinese Medicines' Authentication. Based on the research progress and major achievements acquired in research of each methodology including identification of origin, description, microscopic, physical, chemical and biological characteristics of Chinese medicines, it is expounded that the development process of each methodology combined modem digital technology, information science and its own characteristics. And the development direction is further described for methodology of Chinese Medicines' Authentication towards systematization and informationization.
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Química Farmacêutica/história , Química Farmacêutica/métodos , Contaminação de Medicamentos/prevenção & controle , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/história , Medicamentos de Ervas Chinesas/farmacologia , História do Século XX , História do Século XXI , Controle de QualidadeRESUMO
The aim of this paper is to apply near infrared spectroscopy techniques to construct a rapid identification method for 8 kinds of mineral Chinese Medicines containing carbonates. The qualitative model using clustering analysis method in OPUS software can identify accurately 8 kinds of carbonate-containing mineral Chinese medicines. The near-infrared quantitative model was established by using partial least squares method (PLS) for 7 mineral Chinese Medicines in which main component is calcium carbonate. Compared with the results by EDTA titration, the established quantitative analysis model for calcium carbonate content showed a good prediction result that when the content is between 47.61% -99.17%, the average relative deviation of the prediction result is 0.24% and the average recovery rate was 100.3%. The results also showed that the model using near infrared spectroscopy can get not only a rapid identification of the 8 mineral Chinese medicines containing carbonates, but also an accurate and reliabe content determination of calcium carbonate for the 7 mineral Chinese medicines which contain the component.
Assuntos
Carbonatos/análise , Medicina Tradicional Chinesa , Minerais/química , Espectrofotometria Infravermelho/métodos , Software , Fatores de TempoRESUMO
OBJECTIVE: To identify Hibiscus syriacus and its adulterants using DNA barcoding technique. METHODS: Nine samples of five species were PCR amplified and sequenced, and twelve samples were downloaded from the GenBank. The intra-specific and interspecific K2P distances were calculated, and neighbor-joining( NJ) tree was constructed by MEGA 5.0. RESULTS: The results showed the intra-specific genetic distances of Hibiscus syriacus were ranged from 0.009 to 0.056, which were far lower than inter-specific genetic distances between Hibiscus syriacus and its adulterants (0.236 - 0.301). Variable sites within Hibiscus syriacus ranged from 2 to 9 which were far less than the adulterants (45 - 52); Different samples of Hibiscus syriacus were gathered together and could be distinguished from its adulterants by NJ tree. CONCLUSION: ITS2 can discriminate Hibiscus syriacus from its adulterants correctly. The ITS2 region is an efficient barcode for authentication of Hibiscus syriacus and its adulterants.
Assuntos
Código de Barras de DNA Taxonômico/métodos , DNA de Plantas/genética , DNA Espaçador Ribossômico/genética , Medicamentos de Ervas Chinesas/classificação , Hibiscus/genética , Contaminação de Medicamentos/prevenção & controle , Medicamentos de Ervas Chinesas/química , Flores/classificação , Flores/genética , Hibiscus/classificação , Dados de Sequência Molecular , Filogenia , Folhas de Planta/classificação , Folhas de Planta/genética , Controle de QualidadeRESUMO
Forkhead Box Protein 3 (FoxP3) was identified as a key transcription factor to the occurring and function of the regulatory T cells (Tregs). However, limited evidence indicated its function in tumor cells. To elucidate the precise roles and underlying molecular mechanism of FoxP3 in gastric cancer (GC), we examined the expression of FoxP3 and the consequences of interfering with FoxP3 gene in human GC cell lines, AGS and MKN45, by multiple cellular and molecular approaches, such as immunofluorescence, gene transfection, CCK-8 assay, clone formation assay, TUNEL assay, Flow cytometry, immunoassay and quantities polymerase chain reaction (PCR). As a result, FoxP3 was expressed both in nucleus and cytoplasm of GC cells. Up-regulation of FoxP3 inhibited cell proliferation and promoted cell apoptosis. Overexpression of FoxP3 increased the protein and mRNA levels of proapoptotic molecules, such as poly ADP-ribose polymerase1 (PARP), caspase-3 and caspase-9, and repressed the expression of antiapoptotic molecules, such as cellular inhibitor of apoptosis-1 (c-IAP1) and the long isoform of B cell leukemia/lymphoma-2 (Bcl-2). Furthermore, silencing of FoxP3 by siRNA in GC cells reduced the expression of proapoptotic genes, such as PARP, caspase-3 and caspase-9. Collectively, our findings identify the novel roles of FoxP3 in inhibiting proliferation and inducing apoptosis in GC cells by regulating apoptotic signaling, which could be a promising therapeutic approach for gastric cancer.
Assuntos
Apoptose/fisiologia , Proliferação de Células , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Apoptose/genética , Caspases/genética , Caspases/metabolismo , Linhagem Celular Tumoral , Fatores de Transcrição Forkhead/genética , Humanos , Proteínas Mitocondriais/metabolismo , RNA Interferente Pequeno/genética , Transdução de Sinais , Regulação para CimaRESUMO
OBJECTIVE: To study the quality standard of Selaginella moellendorffii. METHODS: The macroscopic and microscopic characteristics were observed, amentoflavone was used as reference substance in the TLC identification and HPLC method was used to determine the content of amentoflavone. RESULTS: The TLC method used GF254 taking toluene-ethyl formate-formic acid (5: 4: 0. 5) as the developer and ethanol solution of ferric chloride as the chromogenic reagent with the results that it appeared four clear spots. The HPLC method took amentoflavone as the reference substance, and acetonitrile-water (containing 3% tetrahydrofuran and 3% trifluoroacetic acid) as the mobile phase; Detection wavelength was 330 nm. The contents of samples from 3 different batches were determined, and the lowest content of amentoflavone was 0.4%. CONCLUSION: This method is reliable and accurate, which can be used for the quality control of Selaginella moellendorfii.
Assuntos
Biflavonoides/análise , Cromatografia em Camada Fina/métodos , Medicamentos de Ervas Chinesas/química , Selaginellaceae/anatomia & histologia , Selaginellaceae/química , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/normas , Folhas de Planta/anatomia & histologia , Folhas de Planta/química , Caules de Planta/anatomia & histologia , Caules de Planta/química , Controle de Qualidade , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To identify Peucedani Radix and its adulterants using DNA barcoding technique. METHODS: Total genomic DNA was isolated from Peucedani Radix and its adulterants. Nuclear DNA ITS2 sequences were amplified and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.0. The Kimura 2-Parameter(K2P) distances were calculated using software MEGA 4. 0. Identification analyses were performed using BLAST1, Nearest Distance and Neighbor-Joining (NJ) methods, and the secondary structure of the ITS2 sequence differences between species were analyzed. RESULTS: Different samples of Peucedani Radix were gathered together and distinguished from its adulterants by NJ tree. The ITS2 secondary structure showed that Peucedani Radix could be differentiated obviously from its adulterants. CONCLUSION: ITS2 sequence is able to identify Peucedani Radix and its adulterants correctly, which provides a scientific basis for fast and accurate identification of the herb.
Assuntos
Apiaceae/genética , Código de Barras de DNA Taxonômico , DNA de Plantas/genética , DNA Espaçador Ribossômico/genética , Contaminação de Medicamentos , Apiaceae/classificação , Sequência de Bases , Dados de Sequência Molecular , Filogenia , Raízes de Plantas/genética , Plantas Medicinais/classificação , Plantas Medicinais/genética , Reação em Cadeia da Polimerase , Controle de Qualidade , Especificidade da EspécieRESUMO
Sepsis is a syndrome with poor prognosis. Nucleotide-binding domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome and T helper 17 (Th17) cells are involved in the pathogenesis of inflammatory diseases. This study aims to explore their roles and underlying mechanisms in sepsis. The blood and bronchoalveolar lavage fluid are collected from sepsis patients and healthy donors. A sepsis mice model is established by cecal ligation puncture (CLP). The contents of cytokines are detected by ELISA. The amounts of Th17 cells, IL-17A, IL-1ß, IL-18, and lipopolysaccharide is significantly elevated in sepsis patients. The increased differentiation of Th17 cells can promote lung cell pyroptosis and induce hyperpermeability via activating NLRP3 inflammasome and p38 pathway. The inhibitors targeting Th17 cells, NLRP3 inflammasome, and p38 pathway can significantly alleviate lung injury in sepsis mice. Th17 cells can secrete IL-17A to activate NLRP3 inflammasome via p38 signaling pathway, which contributes to the development of sepsis-induced acute lung injury.
Assuntos
Células Epiteliais Alveolares , Inflamassomos , Sepse , Células Th17 , Humanos , Sepse/imunologia , Sepse/metabolismo , Sepse/patologia , Células Th17/imunologia , Células Th17/patologia , Lipopolissacarídeos/sangue , Interleucina-17/sangue , Interleucina-1beta/sangue , Interleucina-18/sangue , Piroptose , Permeabilidade da Membrana Celular , Transdução de Sinais , Células A549 , Inflamassomos/metabolismo , Animais , Camundongos , Modelos Animais de Doenças , Células Epiteliais Alveolares/metabolismo , Células Epiteliais Alveolares/patologiaRESUMO
Kidney transplantation (KT) is an ultimate treatment of end-stage chronic kidney disease, which can meet a lot of complications induced by immune system. With under-controlled immunosuppression, the patient will obtain a good prognosis. Otherwise, allograft disfunction will cause severe organ failure and even immune collapse. Acute or chronic allograft dysfunction after KT is related to Th17, Treg, and Th17/Treg to a certain extent. Elevated Th17 levels may lead to acute rejection or chronic allograft dysfunction. Treg mainly plays a protective role on allografts by regulating immune response. The imbalance of the two may further aggravate the balance of immune response and damage the allograft. Controlling Th17 level, improving Treg function and level, and adjusting Th17/Treg ratio may have positive effects on longer allograft survival and better prognosis of receptors.
Assuntos
Transplante de Rim , Humanos , Linfócitos T Reguladores , Células Th17 , Imunidade , ImunomodulaçãoRESUMO
BACKGROUND: To study the expression of the RIZ1 (Retinoblastoma protein-interacting zinc-finger gene 1) gene and investigate the promoter region methylation status of RIZ1 gene in the human esophageal squamous cell carcinoma (ESCC) cell lines of KYSE150, KYSE510, TE13, EC9706, CaEsl7, and EC109. To investigate the influence of DNMT (DNA methyltransferase) 5-aza-CdR(5-aza-2'-deoxycytidine) on the transcription of the RIZ1 gene in one cell line whose RIZ1 gene promoter region methylation was detected, and to investigate its influence on the cell proliferation. METHODS: Real-time PCR (Real-time quantitative PCR) and an immunohistochemistry technique was used to get the expression of RIZ1 in specimens from 6 human ESCC cell lines and 28 ESCC patients (tumor tissues and adjacent non-cancerous tissues). MSP (Methylation-specific PCR) was used to investigate the promoter region methylation status of the RIZ1 gene in the 6 ESCC cell lines. One cell line, whose RIZ1 gene promoter region methylation was detected, was chosen for the next studies in which it was treated it by with 5-aza-CdR. Real-time PCR was used to investigate its influence on the transcription of RIZ1 gene and MTT (methyl thiazolyl tetrazolium) was used to detect if 5-aza-CdR inhibits the proliferation of the cell line. RESULTS: In the 28 ESCC patient samples, RIZ1 expression was significantly lower in the tumor tissues than that in their adjacent non-cancerous tissues (p < 0.05). Consistently, immunohistochemistry analyses of RIZ1 protein expression showed that in the ESCC tissues RIZ1 protein expression was also significantly lower than in the adjacent tissues. In the human ESCC tissues the rate of expression accounts for 0% (0/12), and in the adjacent noncancerous tissues the rate of expression was 66.7% (8/12), the correlation was highly significant (chi2 = 12.000, p < 0.05). Promoter methylation of the RIZ1 gene was detected in TE13, CaEsl7, EC109. The cell line TE13 was chosen for the next studies. The expression of RIZ1 mRNA in TE-13 was up-regulated after having been treated with 5-aza-CdR. 5-aza-CdR inhibited cell proliferation of TE-13 in a time and concentration-dependent manner. CONCLUSIONS: Promoter methylation may play an important role in the epigenetic silencing of RIZ1 gene expression. Methylation of the RIZ1 promoter and loss of RIZ1 expression in human ESCC are independent biomarkers. Their determination may offer guidance for selecting appropriate diagnoses and treatments. RIZ1 may be a potential tumor suppressor in human ESCC.
Assuntos
Carcinoma de Células Escamosas/genética , Metilação de DNA , Proteínas de Ligação a DNA/genética , Neoplasias Esofágicas/genética , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Histona-Lisina N-Metiltransferase/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Antimetabólitos Antineoplásicos/farmacologia , Azacitidina/análogos & derivados , Azacitidina/farmacologia , Sequência de Bases , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Metilação de DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Decitabina , Ensaios de Seleção de Medicamentos Antitumorais , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Histona-Lisina N-Metiltransferase/metabolismo , Humanos , Dados de Sequência Molecular , Proteínas Nucleares/metabolismo , RNA Mensageiro/metabolismo , Fatores de Transcrição/metabolismoRESUMO
To investigate the chemical constituents of Chamaecyparis obtusa var. breviramea f. crippsii, various column chromatography and spectroscopic methods were used for the isolation and elucidation of compounds. One new monoterpenoid glucoside, (4S)-4-isopropylcyclohex-l-enecarboxylic acid 4-O-beta-D-glucopyranoside (1), together with five known compounds, (4R)-p-menth-l-ene-7, 8-diol 7-O-beta-D-glucopyranoside (2), skimmin (3), 7-[[6-O-(6-deoxy-alpha-L-mannopyranosyl)-beta-D-glucopyranosyl]oxy]-2H-1-benzopyran-2-one (4), stigmast-4-en-3-one (5) and 1, 4-benzenedicarboxylic acid 1-butyl-4-(2-methylpropyl) ester (6) were isolated and identified from the twigs of this plant. All compounds were isolated from this plant for the first time. The methanol extract of this plant showed cytotoxicity on cancer cell lines A549, BGC-823, Du145 and MDA-MB-231 with IC50 values of 0.94, 1.07, 0.95 and 0.96 microg x mL(-1), respectively. Yet, compounds 1, 2 and 3 showed no cytotoxicity on cancer cell lines HeLa, BGC-823 and A549.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Chamaecyparis/química , Glucosídeos/isolamento & purificação , Monoterpenos/isolamento & purificação , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Cumarínicos/química , Cumarínicos/isolamento & purificação , Cumarínicos/farmacologia , Glucosídeos/química , Glucosídeos/farmacologia , Humanos , Estrutura Molecular , Monoterpenos/química , Monoterpenos/farmacologia , Folhas de Planta/química , Caules de Planta/química , Plantas Medicinais/química , Estigmasterol/análogos & derivados , Estigmasterol/química , Estigmasterol/isolamento & purificação , Estigmasterol/farmacologiaRESUMO
OBJECTIVE: To compare the effect of combination of intradermal needling with oral motor therapy and simple oral motor therapy on salivation in children with cerebral palsy. METHODS: A total of 60 children with salivation in cerebral palsy were randomized into an observation group and a control group, 30 cases in each group. The observation group was treated with intradermal needling (kept for 24 hours each time at Jiache [ST 6], Dicang [ST 4], tongue three needles, etc. ) and oral motor therapy, while the control group was only given oral motor therapy. The intradermal needling was performed 3 times a week, and oral motor therapy was performed 5 times a week, 4 weeks as a course, totally 3 courses of treatment were required. The classification of teacher drooling scale (TDS), drooling severity and Kubota water swallow test, dysphagia disorders survey (DDS) score were compared before treatment and after 4, 8 and 12 weeks of treatment in both groups, and the clinical efficacy was evaluated. RESULTS: After 8 weeks of treatment in the observation group and after 12 weeks of treatment in the two groups, the classification of TDS and drooling severity were improved (P<0.05), and the observation group was better than the control group after 12 weeks of treatment (P<0.05). After 8 and 12 weeks of treatment, the DDS scores of oral period in the observation group were lower than those before treatment (P<0.05). The total effective rate in the observation group was 83.3% (25/30), which was higher than 53.3% (16/30) in the control group (P<0.05). CONCLUSION: The combination of intradermal needling with oral motor therapy can improve salivation symptoms and swallowing function in children with cerebral palsy, the effect is better than oral motor therapy alone, and the effect is earlier.