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This study explored the feasibility of mineral element content and ratios of nitrogen isotopes to discriminate the cultivation mode of Dendrobium nobile in order to provide theoretical support for the discrimination of the cultivation mode of D. nobile. The content of 11 mineral elements(N, K, Ca, P, Mg, Na, Fe, Cu, Zn, Mn, and B) and nitrogen isotope ratios in D. nobile and its substrate samples in three cultivation methods(greenhouse cultivation, tree-attached cultivation, and stone-attached cultivation) were determined. According to the analysis of variance, principal component analysis, and stepwise discriminant analysis, the samples of different cultivation types were classified. The results showed that the nitrogen isotope ratios and the content of elements except for Zn were significantly different among different cultivation types of D. nobile(P<0.05). The results of correlation analysis showed that the nitrogen isotope ratios, mineral element content, and effective component content in D. nobile were correlated with the nitrogen isotope ratio and mineral element content in the corresponding substrate samples to varying degrees. Principal component analysis can preliminarily classify the samples of D. nobile, but some samples overlapped. Through stepwise discriminant analysis, six indicators, including δ~(15)N, K, Cu, P, Na, and Ca, were screened out, which could be used to establish the discriminant model of D. nobile cultivation methods, and the overall correct discrimination rates after back-substitution test, cross-check, and external validation were all 100%. Therefore, nitrogen isotope ratios and mineral element fingerprints combined with multivariate statistical analysis could effectively discriminate the cultivation types of D. nobile. The results of this study provide a new method for the identification of the cultivation type and production area of D. nobile and an experimental basis for the quality evaluation and quality control of D. nobile.
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Dendrobium , Minerais , Análise Discriminante , Análise Multivariada , Isótopos de NitrogênioRESUMO
OBJECTIVE@#To ascertain the influence of light intensity and water content of medium on the total dendrobine of Dendrobium nobile (D. nobile).@*METHOD@#The principal component analysis combined with total dendrobine accumulation was conducted to assess the yield and quality of D. nobile in all treatments. In the experiment, D. nobile plants were cultivated in greenhouse as tested materials, and complete test of 9 treatments was adopted with relative light intensities 75.02%, 39.74%, 29.93% and relative water content of medium 50%, 65%, 80%. The plants were treated in June and harvested till December. Indexes including agronomic traits, fresh weight and dry weight of stem and leaf, ash content, extract, and dendrobine were measured.@*RESULTS@#Under the light intensity treatments of 75.02% with 50%, 65%, 80% water content of medium, the basal stems of plants were comparatively thicker with more leaves, and the fresh weight and dry weight of stems and leaves were significantly higher than other 6 treatments. Leaves in all treatments contained dendrobine. Under the light intensity treatments of 75.02% with 50%, 65%, 80% water content of medium, dendrobine content of leaves was lower while dendrobine contents of other treatments were more than 0.60%. After comprehensive assessment through the principal component analysis and total dendrobine accumulation, the results showed that 3 treatments with relative light intensity of 75.02% ranked the top three.@*CONCLUSIONS@#In brief, the moderately strong light intensity and water content of medium from low to medium can facilitate the growth and yield of D. nobile plants, while light intensity from moderately weak to weak can enhance the dendrobine content.
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Objective To ascertain the influence of light intensity and water content of medium on the total dendrobine of Dendrobium nobile (D. nobile). Method The principal component analysis combined with total dendrobine accumulation was conducted to assess the yield and quality of D. nobile in all treatments. In the experiment, D. nobile plants were cultivated in greenhouse as tested materials, and complete test of 9 treatments was adopted with relative light intensities 75.02%, 39.74%, 29.93% and relative water content of medium 50%, 65%, 80%. The plants were treated in June and harvested till December. Indexes including agronomic traits, fresh weight and dry weight of stem and leaf, ash content, extract, and dendrobine were measured. Results Under the light intensity treatments of 75.02% with 50%, 65%, 80% water content of medium, the basal stems of plants were comparatively thicker with more leaves, and the fresh weight and dry weight of stems and leaves were significantly higher than other 6 treatments. Leaves in all treatments contained dendrobine. Under the light intensity treatments of 75.02% with 50%, 65%, 80% water content of medium, dendrobine content of leaves was lower while dendrobine contents of other treatments were more than 0.60%. After comprehensive assessment through the principal component analysis and total dendrobine accumulation, the results showed that 3 treatments with relative light intensity of 75.02% ranked the top three. Conclusions In brief, the moderately strong light intensity and water content of medium from low to medium can facilitate the growth and yield of D. nobile plants, while light intensity from moderately weak to weak can enhance the dendrobine content.
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<p><b>OBJECTIVE</b>To study the content of mineral elements and the principal components in Gastrodia elata.</p><p><b>METHOD</b>Mineral elements were determined by ICP and the data was analyzed by SPSS.</p><p><b>RESULT</b>K element has the highest content-and the average content was 15.31 g x kg(-1). The average content of N element was 8.99 g x kg(-1), followed by K element. The coefficient of variation of K and N was small, but the Mn was the biggest with 51.39%. The highly significant positive correlation was found among N, P and K . Three principal components were selected by principal components analysis to evaluate the quality of G. elata. P, B, N, K, Cu, Mn, Fe and Mg were the characteristic elements of G. elata.</p><p><b>CONCLUSION</b>The content of K and N elements was higher and relatively stable. The variation of Mn content was biggest. The quality of G. elata in Guizhou and Yunnan was better from the perspective of mineral elements.</p>
Assuntos
Medicamentos de Ervas Chinesas , Gastrodia , Química , Minerais , Análise de Componente PrincipalRESUMO
In order to get to know the imitation of wild Gastrodia elata in life history and phenology period, by G. elata f. elata forest wild simulated cultivation in Dafang county, Guizhou province, observing and recording its morphological characteristics of each growth and development stage. This experiment summarized the law of its life history over 24 months, amplified the characteristics of each 5 phenology periods over the sexual and asexual reproduction of wild simulated cultivated G. elata f. elata in Guizhou. Which the results could clear the process of wild simulated cultivated G. elata f. elata in Guizhou, and provide a theoretical support for the standard technical of the simulated wild G. elata.
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Gastrodia , Fisiologia , Estágios do Ciclo de Vida , Fisiologia , ReproduçãoRESUMO
<p><b>OBJECTIVE</b>To investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.</p><p><b>METHODS</b>Affymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.</p><p><b>RESULTS</b>Compared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.</p><p><b>CONCLUSIONS</b>HepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.</p>
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Humanos , Carcinoma de Células de Transição , Genética , Patologia , Proteínas de Ciclo Celular , Metabolismo , Linhagem Celular Tumoral , Ciclina D1 , Metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , Fisiologia , Proteínas Nucleares , Metabolismo , Proteínas Serina-Treonina Quinases , Metabolismo , Proteínas , Genética , Fisiologia , Neoplasias da Bexiga Urinária , Genética , PatologiaRESUMO
<p><b>BACKGROUND</b>Transforming growth factor-β1 (TGF-β1) is known to be a key fibrogenic cytokine in a number of chronic fibrotic diseases, including chronic allograft nephropathy. We examined the effects of inhibition of TGF-β1 expression by RNA interference on renal allograft fibrosis, and explored the mechanisms responsible for these effects.</p><p><b>METHODS</b>A Sprague-Dawley-to-Wistar rat model of accelerated kidney transplant fibrosis was used. Sixty recipient adult Wistar rats were randomly divided into four groups: group J (sham-operated group), group T (plasmid-transfected group), group H (control plasmid group), and group Y (transplant only group). Rats in group T were transfected with 200 µg of TGF-β1 short hairpin RNA (shRNA). Reverse transcription-polymerase chain reaction and Western blotting were used to examine the expression of TGF-β1, Smad3/7, E-cadherin, and type I collagen. The distribution of type I collagen was measured by immunohistochemistry. The pathologic changes and extent of fibrosis were assessed by hematoxylin and eosin and Masson staining. E-cadherin and α-smooth muscle actin immunohistochemical staining were used to label tubular epithelial cells and fibroblasts, respectively.</p><p><b>RESULTS</b>Plasmid transfection significantly inhibited the expression of TGF-β1, as well as that of its target gene, type I collagen (P < 0.05 and P < 0.01, respectively). In addition, the degree of fibrosis was mild, and its development was delayed in plasmid-transfected rats. In contrast, TGF-β1-shRNA transfection maintained the expression of E-cadherin in tubular epithelial cells while it inhibited the transformation from epithelial cells to fibroblasts. Blood urea nitrogen and serum creatinine were lower in the plasmid group than in the control groups (P < 0.05 and P < 0.01, respectively).</p><p><b>CONCLUSIONS</b>This study suggests that transfection of a TGF-β1-shRNA plasmid could inhibit the fibrosis of renal allografts. The mechanism may be associated with the downregulation of Smad3 and upregulation of Smad7, resulting in suppressed epithelial-myofibroblast transdifferentiation and extracellular matrix synthesis.</p>
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Animais , Ratos , Western Blotting , Transdiferenciação Celular , Genética , Fisiologia , Células Epiteliais , Biologia Celular , Fibrose , Rim , Metabolismo , Patologia , Transplante de Rim , Métodos , Miofibroblastos , Biologia Celular , RNA Interferente Pequeno , Genética , Fisiologia , Ratos Sprague-Dawley , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Crescimento Transformador beta1 , Genética , Metabolismo , Transplante HomólogoRESUMO
<p><b>OBJECTIVE</b>To prepare IL-2-anchored and tumor-derived exosomes vaccine, and investigate the antitumor efficiency of the special cytotoxic T-lymphocytes induced by Ex/GPI-IL-2.</p><p><b>METHODS</b>To construct pEGFP-N1-IL2gpi plasmid coding a fusion gene of a DNA oligo encoding GPI-anchor signal sequence attaching to human IL-2 cDNA. Then T24 cell lines stably expressing GPI-IL-2 proteins (T24/GPI-IL-2) were established. Ex/GPI-IL-2 were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and the morphology and molecule markers were analyzed. The mixed lymphocyte reaction study and cytotoxic study were performed to determine the proliferative effect of T lymphocytes and the cytotoxicity induced by Ex/GPI-IL-2.</p><p><b>RESULTS</b>The pEGFP-N1-IL2gpi plasmid was successfully constructed, and cell lines stably expressing GPI-IL-2 fusion proteins were established. Ex/GPI-IL-2 were small vesicular and saucer-shaped in diameter of 30-90 nm, containing heat shock protein 70, intercellular adhesion molecule-1, MAGE-1 and GPI-IL-2. Ex/GPI-IL-2-pulsed could dendritic cells induce proliferation of T cells and cytotoxic immune response more efficiently (P<0.05).</p><p><b>CONCLUSIONS</b>GPI-IL-2 gene-modified tumor cells can make the exosomes containing GPI-IL-2 with an increased anti-tumor effect. Our study provides a feasible approach for exosome-based tumor immunotherapy of bladder transitional cell tumors.</p>
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Humanos , Vacinas Anticâncer , Alergia e Imunologia , Carcinoma de Células de Transição , Alergia e Imunologia , Metabolismo , Patologia , Linhagem Celular Tumoral , Proliferação de Células , Exossomos , Genética , Metabolismo , Glicosilfosfatidilinositóis , Metabolismo , Proteínas de Choque Térmico HSP70 , Metabolismo , Molécula 1 de Adesão Intercelular , Metabolismo , Interleucina-2 , Metabolismo , Antígenos Específicos de Melanoma , Metabolismo , Plasmídeos , Proteínas Recombinantes de Fusão , Metabolismo , Linfócitos T Citotóxicos , Alergia e Imunologia , Transfecção , Neoplasias da Bexiga Urinária , Alergia e Imunologia , Metabolismo , PatologiaRESUMO
<p><b>OBJECTIVE</b>To prepare a vaccine of IL-12-anchored exosomes derived from renal cancer cells and to evaluate its antitumor effect in vitro.</p><p><b>METHODS</b>A mammalian co-expression plasmid of glycolipid-anchor-IL-12 (GPI-IL-12) was constructed by subcloning IL-12A chain gene (P35 subunit) and a fusion gene containing GPI-anchor signal sequence and IL-12B chain gene (P40 subunit) in pBudCE4.1. Confocal laser scanning microscopy and flow cytometry were used to analyze the expression of the fusion proteins. Transmission electron microscopy and Western blot were used to identify the morphology and characteristic molecules of exosomes separated by ultrafiltration and sucrose gradient centrifugation. The function of IL-12-anchored exosomes was determined by IFN-gamma release assay.</p><p><b>RESULTS</b>Mammalian co-expression plasmids were successfully constructed. Confocal laser scanning microscopy and flow cytometric analysis of the RC-2-GPI-IL-12 transfectants showed the expression of IL-12 on the cell surface. Exosomes were purified by ultrafiltration and sucrose gradient centrifugation, which were 30-80 nm in diameter, typically saucer-shaped, and expressing HSP70, ICAM-1, G250 and GPI-IL-12. (80.0 +/- 9.6) pg/ml of IL-12 was detected in 10 microg/ml exosomes and it significantly induced the release of IFN-gamma. Stimulation with EXO-IL-12 could efficiently induce antigen-specific cytotoxic T lymphocytes (CTL), resulting in more significant cytotoxic effects in vitro.</p><p><b>CONCLUSION</b>A vaccine of exosomes-GPI-IL-12 can be obtained from the culture supernatant of renal cancer cells modified to express anchored IL-12. This vaccine expressing IL-12 and tumor associated antigen G250 may become a new strategy for the treatment of renal cancer.</p>
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Humanos , Antígenos de Neoplasias , Metabolismo , Vacinas Anticâncer , Alergia e Imunologia , Anidrase Carbônica IX , Anidrases Carbônicas , Metabolismo , Linhagem Celular Tumoral , Citotoxicidade Imunológica , Exossomos , Genética , Metabolismo , Glicosilfosfatidilinositóis , Genética , Metabolismo , Proteínas de Choque Térmico HSP70 , Metabolismo , Molécula 1 de Adesão Intercelular , Metabolismo , Interferon gama , Secreções Corporais , Interleucina-12 , Genética , Metabolismo , Neoplasias Renais , Metabolismo , Patologia , Plasmídeos , Linfócitos T Citotóxicos , Biologia Celular , Alergia e Imunologia , TransfecçãoRESUMO
<p><b>OBJECTIVE</b>To isolate and purify exosomes derived from human bladder transitional cell carcinoma T24 cells, analyze the morphology and protein composition, and investigate the antitumor effect of specific cytotoxic T lymphocytes induced by exosomes.</p><p><b>METHODS</b>Exosomes were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and characterized by electron microscopy and Western blot. Dendritic cells were amplified and purified from peripheral blood and pulsed with exosomes. Then they were co-cultured with T cells, and divided into 3 groups: exosome-pulsed DC group, unplused DC group and control group. Alamar-Blue assay was used to evaluate the specific cytolytic activity.</p><p><b>RESULTS</b>The exosomes were in size about 30 approximately 90 nm saucer-shaped membranous vesicles. HSP70, ICAM-1 and CK20 were detected by Western blot. The CTL induced by DC pulsed with exosomes had significant cytolytic activity (P < 0.01).</p><p><b>CONCLUSION</b>The exosomes derived from T24 cells are loaded with immunoprotein HSP70 and ICAM-1, and DC pulsed with exosomes can promote the anti-tumor effect of CTLs in vitro.</p>
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Humanos , Carcinoma de Células de Transição , Patologia , Linhagem Celular Tumoral , Técnicas de Cocultura , Citotoxicidade Imunológica , Alergia e Imunologia , Células Dendríticas , Biologia Celular , Alergia e Imunologia , Exossomos , Alergia e Imunologia , Metabolismo , Proteínas de Choque Térmico HSP70 , Metabolismo , Molécula 1 de Adesão Intercelular , Metabolismo , Queratina-20 , Metabolismo , Ativação Linfocitária , Linfócitos T , Biologia Celular , Alergia e Imunologia , Linfócitos T Citotóxicos , Alergia e Imunologia , Neoplasias da Bexiga Urinária , PatologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the proliferation, apoptosis and mechanisms on T24 cell of transitional cell carcinoma of bladder (TCCB) by crocin.</p><p><b>METHOD</b>MTT assay was used to evaluate the proliferation of T24 cells. The changes of cell cycle and cell apoptotic percentage were measured by flow cytometry. T24 cells were inoculated into BALB/c nude mice to establish model of carcinoma of bladder. The mice were randomly divided into control group and experimental group. After treatment with 50 mmol x L(-1) crocin, the inhibited growth of tumor was observed. Electronic microscope was used to observe the morphological changes. The expressions of Bcl-2, Bax, Survivin and Cyclin D1 were detected by immunohistochemistry.</p><p><b>RESULT</b>The growth of T24 cells was remarkably inhibited after treatment of crocin. Flow cytometric profiles revealed that crocin led to the increase of the cells in G0/G1 phase, the percentage of cell apoptosis was also increased. Crocin could inhibit the growth of BALB/c xenograft tumor. The morphology changes of cell apoptosis were observed. Bcl-2, Cyclin D1 and survivin expressions determined by immunohistochemical staining were down-regulated after treatment with Bax expression up-regulated.</p><p><b>CONCLUSION</b>Crocin exerts both in vitro and in vivo anti-cancer effect on TCCB T24 cell line. The mechanisms may change tumour cell cycle and induce tumour cell apoptosis by down-regulating the expression of Bcl-2, Survivin, Cyclin D1 and up-regulating the expression of Bax.</p>
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Animais , Humanos , Camundongos , Apoptose , Carcinoma de Células de Transição , Tratamento Farmacológico , Patologia , Carotenoides , Farmacologia , Usos Terapêuticos , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Ciclina D1 , Metabolismo , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Proteínas Inibidoras de Apoptose , Camundongos Endogâmicos BALB C , Camundongos Nus , Microscopia Eletrônica de Transmissão , Proteínas Associadas aos Microtúbulos , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Proteínas Repressoras , Transplante Heterólogo , Neoplasias da Bexiga Urinária , Tratamento Farmacológico , PatologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the changes of gene expression profile in transitional cell carcinoma of bladder T24 cell after crocin treatment, in order to find the possible crocin targets.</p><p><b>METHOD</b>The bladder cancer T24 cell line was treated with crocin. MTT assay was adopted to determine the inhibition rate for selecting the best effect time and concentration of crocin. Differentially expressed genes on groups with or without treatment of crocin were screened with high throughout cDNA microarray. One up-regulated gene p21(WAF1) and one down-regulated gene cyclinD1 were selected to undergo analysis by the reverse transcription polymerase chain reaction (RT-PCR). Moreover, immunocytochemical method was used to evaluate p21(WAF1) and cyclinD1 protein expression.</p><p><b>RESULT</b>The growth of T24 cells was inhibited remarkably following a marked positive correlation between crocin concentration, time and inhibitor rate. When 3 mmol x L(-1) crocin treated T24 cells for 48h, the difference was significant compared with the control group (P < 0.05). Crocin induced wide changes of the gene expression profile of T24 cells. A total of 836 genes were up-regulated or down-regulated by more than 2 times, which were involved cell cycle controlling, DNA cell apoptosis, replication factor, and so on. The mRNA expression of p21(WAF1) and cyclinD1 detected by RT-PCR were in accordance with cDNA microarray data. The results of immunocytochemical method showed that p21(WAF1) and cyclinD1 protein expression were consistent with those mRNA expression.</p><p><b>CONCLUSION</b>Crocin can induce the significant alteration of gene expression profile of T24 cell. It is suggested that the widly konwn anti-tumor effects of crocin are medicated at least in part by regulating the cell cycle controlling gene expression.</p>
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Humanos , Antineoplásicos , Farmacologia , Carotenoides , Farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Relação Dose-Resposta a Droga , Regulação para Baixo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Regulação para Cima , Neoplasias da Bexiga Urinária , Metabolismo , PatologiaRESUMO
<p><b>OBJECTIVE</b>To study the effect and mechanism of saffor injection on renal ischemia/reperfusion (I/R) injury in rats.</p><p><b>METHOD</b>Seventy-five SD rats were randomly divided into five groups (n = 15, in each), normal control groups, I/R control groups, low-dose treatment groups, middle-dose treatment groups and high-dose treatment groups. After rat's I/R injury model was established, renal function was assessed by measuring serum creatinine, blood urea nitrogen, urine osmotic pressure and urine osmotic pressure/blood osmotic pressure, the apoptosis rate in I/R renal tissure was measured by TUNEL method and caspase-3 concentration was measured by immunohistochemistry.</p><p><b>RESULT</b>Reperfusion of the ischemic kidney induced marked renal dysfunction. Saffor injection significantly inhibited the reperfusion-associated increase in apoptosis rate and caspase-3 protein absorbance value. Moreover, the renal dysfunction at all treatment groups was markedly ameliorated by Saffor injection. (P < 0.01).</p><p><b>CONCLUSION</b>The results show that saffor injection significantly reduces the renal dysfunction and injury caused by I/R of the kidney, And the protective effect of Saffor injection may be related to the inhibition of cell apoptosis and caspase-3 gene expression following renal I/R.</p>
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Animais , Feminino , Masculino , Ratos , Apoptose , Nitrogênio da Ureia Sanguínea , Carthamus tinctorius , Química , Caspase 3 , Metabolismo , Creatinina , Sangue , Medicamentos de Ervas Chinesas , Farmacologia , Injeções , Rim , Pressão Osmótica , Plantas Medicinais , Química , Distribuição Aleatória , Ratos Sprague-Dawley , Traumatismo por Reperfusão , Sangue , PatologiaRESUMO
<p><b>OBJECTIVE</b>To construct deltaNp63 specific small hairpin RNA (shRNA) expressing plasmid,to examine its inhibitory effect to the expression of deltaNp63 protein and mRNA in transitional cell carcinoma of the bladder (TCCB) , its effect on TCCB cells cycle and proliferation.</p><p><b>METHODS</b>DeltaNp63 specific oligonucleotides were designed and synthesized. These oligonucleotides were annealed to form double strand DNA fragments and this fragment was cloned into Pgenesil-1 plasmid. The recombinant deltaNp63-shRNA expression construct was confirmed by using Pst I + Sal I double digestion and by sequencing. Fluorescence staining was used to confirm the success of transfection in TCCB cells under the fluorescence microscope. The inhibitory effect of deltaNp63-shRNA construct was examined with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining assay. The cell cycle of TCCB cells was assayed by flow cytometry (FCM). The cellular proliferation of TCCB cells was assayed by tetrazolium bromide (MTT) colorimetry.</p><p><b>RESULTS</b>The deltaNp63-shRNA expression plasmid was successfully constructed and transfected into TCCB cells. It can effectively reduce the expression of deltaNp63 protein and mRNA. The reduction rate of deltaNp63 mRNA was 63.0%, and the G0/G1 ratio was increased and S phase was decreased in transfected TCCB cells. The cellular proliferation was also lower in transfected 5637 cells in comparrison with that of non-transfected TCCB cells.</p><p><b>CONCLUSION</b>A deltaNp63-shRNA expression plasmid, constructed from Pgenesil-1 plasmid, can successfully be transfected into TCCB cells and can effectively inhibit the expression of deltaNp63 protein and mRNA. It also can take part in regulation of the cell cycling and inhibit the cellular proliferation of TCCB cells.</p>
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Humanos , Carcinoma de Células de Transição , Genética , Metabolismo , Patologia , Ciclo Celular , Genética , Fisiologia , Linhagem Celular Tumoral , Proliferação de Células , Proteínas de Ligação a DNA , Genética , Fisiologia , Imuno-Histoquímica , Microscopia de Fluorescência , Plasmídeos , Genética , Interferência de RNA , RNA Mensageiro , Genética , RNA Interferente Pequeno , Genética , Proteínas Recombinantes , Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transativadores , Genética , Fisiologia , Fatores de Transcrição , Transfecção , Proteínas Supressoras de Tumor , Genética , Fisiologia , Neoplasias da Bexiga Urinária , Genética , Metabolismo , PatologiaRESUMO
"Clinical Laboratory Diagnosis"is a course of trivial and complicated contents,lacking systematization and consisten- cy.Therefore,the students are not interested in acquiring knowledge eagerly.How to improve the quality and efficiency of teach- ing is concluded as clear focus,constant renewed content and attractive class.