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PURPOSE: This study was designed to evaluate the diagnostic accuracy of computed tomography (CT)-guided biopsy for small lung nodules (SLNs) (≤20 mm) and to assess related complication rates. METHODS: We reviewed the Pubmed, Embase, and Cochrane Library databases to identify all relevant studies published as of April 2020. Random effects modeling were then used to evaluate pooled data pertaining to technical success rates, diagnostic accuracy, pneumothorax rates, and rates of hemoptysis. The meta-analysis was conducted using Stata v12.0. RESULTS: In total, we identified 25 relevant studies for incorporation into this meta-analysis, incorporating 2922 total CT-guided lung biopsy. Pooled technical success rates, diagnostic accuracy, pneumothorax rates, and hemoptysis rates were 94% (95% confidential interval [CI], 0.91-0.98), 90% (95% CI, 0.88-0.93), 19% (95% CI:, 0.15-0.24), and 12% (95% CI, 0.08-0.15), respectively. We observed significant heterogeneity among these studies for all 4 of these parameters (I = 90.0%, 82.7%, 88.6%, and 88.4%, respectively). When we conducted a meta-regression analysis, we did not identify any variables that influenced diagnostic accuracy or technical success, pneumothorax, or hemoptysis rates. Publication bias risk analyses suggested that there was relatively little risk of publication bias pertaining to pneumothorax rates (P = 0.400) or hemoptysis rates (P = 0.377). In contrast, we detected a high risk of publication bias pertaining to reported technical success rates (P = 0.007) and diagnostic accuracy (P = 0.000). CONCLUSIONS: A CT-guided biopsy can be safely and effectively used to diagnose SLNs.
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Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/patologia , Radiografia Intervencionista/métodos , Nódulo Pulmonar Solitário/diagnóstico por imagem , Nódulo Pulmonar Solitário/patologia , Tomografia Computadorizada por Raios X/métodos , Humanos , Biópsia Guiada por Imagem , Pulmão/diagnóstico por imagem , Pulmão/patologia , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To ivestigate the effect of daily walking number on clinical, inflammatory and nutritional indexes for patients with chronic kidney disease. METHODS: 90 non-dialysis patients with chronic kidney disease were selected and randomly divided equally into three groups, for groups A, B and C. 30 patients for group A were asked for the number of daily walk should less than 5 000 steps, while group B patients were asked for 5 000-9 999 steps of walk and group C for more than 10 000 steps. Basic treatments were given for each group of patients and basic information, clinical, inflammatory and nutritional datas of patients were collected. RESULTS: 87 patients with chronic kidney disease completed the study, with baseline data between group A, B, C (n=30, 29, 28) consistently. After 3 months of exercise, there were no significant changes on blood lipids, serum uric acid and parathyroid hormone (PTH) for three groups, with serum creatinine of three groups stably. However, in group C, hemoglobin, ferritin, transferrin saturation were found increased significantly (P<0.05). For nutritional indexes, increasing of albumin and prealbumin level were found in three groups, while significant differences were only found in group B and C (P<0.05) and group C increased most. There were no significant change on body mass index (BMI), upper arm skinfold thickness and SGA score in three groups. For inflammatory data, significant decrease of C-reactive protein (CRP) and interleukin-6 (IL-6) were only seen in group C (P<0.05). CONCLUSION: Walking does not increase the burden of the kidney, but can improve the nutrition and clinical indicators of patients, reduce inflammation.
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Inflamação/terapia , Estado Nutricional , Insuficiência Renal Crônica/sangue , Caminhada , Proteína C-Reativa/análise , Humanos , Inflamação/sangue , Interleucina-6/sangue , Lipídeos/sangue , Hormônio Paratireóideo/sangue , Diálise Renal , Ácido Úrico/sangueRESUMO
The incidence of sexual dysfunction is higher in men with chronic kidney disease (CKD) than in those without, of which ED is the most common clinical manifestation. Sexual dysfunction is closely related to malfunction of the endocrine system in CKD males, mainly including the disorder in the hypothalamus-pituitary-gonadal axis and hyperprolactinemia. Besides, blood vessels, the nervous system, psychological status, trace elements, and drugs are also contributory factors. At present, sexual dysfunction in CKD males is diagnosed chiefly by scale assessment, clinical manifestation and special examination, and its treatment focuses on the correction of the endocrine system malfunction, as by kidney transplantation or by drug therapy with recombinant human erythropoietin, 1,25- (OH)2 D3, bromocriptine, losartan, or zinc preparation. In addition, conventional treatment with phosphodiesterase-5 inhibitors can be used as a supplement. This paper outlines the recent progress in the studies of sexual dysfunction in CKD males, covering its risk factors, etiology, pathophysiology, diagnosis and treatment.
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Insuficiência Renal Crônica , Disfunções Sexuais Fisiológicas , Suplementos Nutricionais , Humanos , Incidência , Masculino , Inibidores da Fosfodiesterase 5/uso terapêutico , Insuficiência Renal Crônica/complicações , Disfunções Sexuais Fisiológicas/complicaçõesRESUMO
Recent researches have reported the extensive pharmacological activities of Ginsenoside Rg1 including antioxidant, anti-inflammatory, and anticancer properties. Furthermore Rg1 was also shown to protect various kinds of cells from self-digestion by its anti-autophagy activity. In previous studies, angiotensin II (Ang II), a key mediator of renin-angiotensin system, has been demonstrated to contribute to the progression of renal injury including abnormal autophagy. However, whether Rg1 can relieve Ang II-induced autophagy in podocyte as well as the underlying molecular mechanism remains to be elucidated. Here, we employed Ang II-treated podocyte as a model to investigate the effect of Rg1 on autophagy and the involved signal pathways. In the present study, we found that Ang II strongly promoted autophagy in immortalized mouse podocyte cells by observing the formation of autophagosomes and detecting the expression of autophagic marker, for example, LC3-II. Notably, compared to the Ang II-treated cells, treatment with Rg1 significantly inhibited the formation of autophagosomes and expression of autophagy-related proteins in Ang II pre-treated podocyte. Meanwhile, Rg1 downregulated the activity of AMPK and GSK-3ß and upregulated the activity of P70S6K in Ang II-treated podocyte. In conclusion, these findings demonstrate that Ang II promotes autophagy in podocyte, and Rg1 effectively attenuates this process through AMPK/mTOR/PI3K pathway, suggesting that Rg1 may be beneficial to alleviate podocyte injury.
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Proteínas Quinases Ativadas por AMP/metabolismo , Angiotensina II/metabolismo , Autofagia/efeitos dos fármacos , Ginsenosídeos/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Podócitos/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Podócitos/citologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
Objective To observe the effect of Modified Dihuang Yinzi Recipe (MDYR) and Hua- tan Tongluo Decoction (HTD) on neurological function of middle cerebral artery occlusion (MCAO) model rats. Methods Forty SD rats were randomly divided into 5 groups, i.e., the sham-operation group, the model group, the nimodipine (NMDP) group, the MDYR group, and the HTD group, 8 in each group. NMDP liquid was administered to rats in the NMDP group by gastrogavage at the daily dose of 12 mg/kg. MDYR liquid was administered to rats in the MDYR group by gastrogavage at the daily dose of 7. 9 mg/kg. HTD liquid was administered to rats in the HTD group by gastrogavage at the daily dose of 6. 5 mg/kg. Equal volume of distilled water as administered to rats in the sham-operation group and the model group by gastrogavage. All intervention lasted for 7 successive days. MCAO rat model was established. The Zealonga neurology score was measured. Neurological function was scored at 3 and 6 h, day 1 , 6, and 7, respectively. Levels of corticotropin releasing hormone ( CRH) in plasma, adrenocorticotropic hormone (ACTH) and cortisone (CORT) in serum were detected using radioimmunoassay. The expression of ma- trix metalloproteinase-9 ( MMP9 ) in brain tissue was detected using immunohistochemical staining. Results Compared with the sham-operation group, the Zea-longa neurological score increased; levels of CRH and ACTH decreased (P <0. 05, P <0. 01) , expression levels of CORT and MMP-9 (in brain tis- sue) increased in the model group (P <0. 01). Compared with the model group, the Zea-longa neurologi- cal score obviously decreased, levels of CRH and ACTH increased, expression levels of CORT and MMP-9 (in brain tissue) decreased (P <0. 05, P <0. 01) in the MDYR group and the HTD group. Com- pared with the NMDP group, serum CORT decreased in the MDYR group (P <0. 05) ; MMP-9 expression level decreased in the HTD group (P <0. 01). Conclusions MDYR and HTD could obviously improve neurologic function of MCAO rats. Its mechanism might be related to MDYR regulating disordered HPA ax- is and HTD inhibiting MMP-9 expression.
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Medicamentos de Ervas Chinesas , Infarto da Artéria Cerebral Média , Doenças do Sistema Nervoso , Animais , Hormônio Liberador da Corticotropina , Medicamentos de Ervas Chinesas/uso terapêutico , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/tratamento farmacológico , Doenças do Sistema Nervoso/tratamento farmacológico , Doenças do Sistema Nervoso/etiologia , Ratos , Ratos Sprague-DawleyRESUMO
While Helicobacter pylori (Hp) infection is closely associated with IgA nephropathy (IgAN), the underlying molecular mechanisms remain to be elucidated. This study was to investigate the effect of cytotoxin associated gene A protein (CagA), a major virulence factor of Hp, on the production and underglycosylation of IgA1 in the B cell line DAKIKI cells. Cells were cultured and treated with recombinant CagA protein. We found that CagA stimulated cell proliferation and the production of IgA1 in a dose-dependent and time-dependent manner. Moreover, CagA promoted the underglycosylation of IgA1, which at least partly attributed to the downregulation of ß1,3-galactosyltransferase (C1GALT1) and its chaperone Cosmc. In conclusion, we demonstrated that Hp infection, at least via CagA, may participate in the pathogenesis of IgAN by influencing the production and glycosylation of IgA1 in B cells.
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Antígenos de Bactérias/farmacologia , Linfócitos B/efeitos dos fármacos , Proteínas de Bactérias/farmacologia , Imunoglobulina A/metabolismo , Antígenos de Bactérias/genética , Antígenos de Bactérias/metabolismo , Linfócitos B/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Galactosiltransferases/genética , Expressão Gênica/efeitos dos fármacos , Glomerulonefrite por IGA/etiologia , Glomerulonefrite por IGA/genética , Glomerulonefrite por IGA/metabolismo , Glicosilação/efeitos dos fármacos , Infecções por Helicobacter/complicações , Infecções por Helicobacter/microbiologia , Helicobacter pylori/fisiologia , Interações Hospedeiro-Patógeno , Humanos , Chaperonas Moleculares/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
A sensitive and reliable ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the simultaneous determination of four active components of Semen Cassiae extract (aurantio-obtusin, chrysoobtusin, obtusin and 1-desmethylobtusin) in rat plasma after oral administration. Chromatographic separation was achieved on an Agilent Poroshell 120 C18 column with gradient elution using a mobile phase that consisted of acetonitrile-ammonium acetate in water (30 mm) at a flow rate of 0.4 mL/min. Detection was performed by a triple-quadrupole tandem mass spectrometer in multiple reaction monitoring mode. The calibration curve was linear over a range of 3.24-1296 ng/mL for aurantio-obtusin, 0.77-618 ng/mL for chrysoobtusin, 34.55-1818 ng/mL for obtusin and 1.86-1485 ng/mL for 1-desmethylobtusin. Inter- and intra-day assay variation was <15%. All analytes were shown to be stable during all sample storage and analysis procedures.
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Antraquinonas/sangue , Cassia/química , Extratos Vegetais/química , Sementes/química , Animais , Antraquinonas/química , Antraquinonas/farmacocinética , Cromatografia Líquida de Alta Pressão/métodos , Modelos Lineares , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrometria de Massas em Tandem/métodosRESUMO
The objective of the study was to evaluate the effects of trace mineral supplementation in sows during gestation and lactation on the performance and health status of sows and their offspring. Sows (n = 30; Landrace × Yorkshire; avg parity = 3.9) were randomly allocated into two dietary treatments. Sows received a basal diet supplemented with 12 mg/kg Cu, 30 mg/kg Fe, 90 mg/kg Zn, 70 mg/kg Mn, 0.30 mg/kg Se, and 1.5 mg/kg I from an inorganic trace mineral source (ITM) or a blend of hydroxychloride and organic trace mineral source (HOTM) from day 1 of gestation until the end of the lactation period at day 21. Compared to the ITM, the HOTM supplementation increased (P < 0.05) both litter birth weight and individual piglet birth weight. Although not statistically significant, HOTM tended to increase (P = 0.069) the level of lactose in colostrum. HOTM increased (P < 0.05) the concentration of Mn and Se in the colostrum, milk, and serum of sows and/or piglets. Notably, the Zn concentration in the serum of sows was higher in sows supplemented with ITM compared to HOTM. Moreover, HOTM increased (P < 0.05) the activities of GPX and SOD in gestating sows and piglets, as well as increased (P < 0.05) cytokines (IL-1ß, TNF-α, and IL-10) in the serum of sows. The immunoglobulins (IgA, IgG, and IgM) also increased in sows and/or piglets at certain experimental time points. In conclusion, HOTM supplementation positively affected piglet development and improved the health status of sows and piglets potentially by regulating redox homeostasis and immunity.
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PURPOSE: The polarization of macrophages with the resulting inflammatory response play a crucial part in tissue and organ damage due to inflammatory. Study has proved Lian Hua Qing Wen capsules (LHQW) can reduce activation of inflammatory response and damage of tissue derived from the inflammatory reactions. However, the mechanism of LHQW regulates the macrophage-induced inflammatory response is unclear. Therefore, we investigated the mechanism of LHQW regulated the inflammatory response of M1 macrophages by cellular experiments and computer simulations. METHODS: This study has analysed the targets and mechanisms of macrophage regulating inflammatory response at gene and protein levels through bioinformatics. The monomeric components of LHQW were analyzed by High Performance Liquid Chromatography (HPLC). We established the in vitro cell model by M1 macrophages (Induction of THP-1 cells into M1 macrophages). RT-qPCR and immunofluorescence were used to detect changes in gene and protein levels of key targets after LHQW treatment. Computer simulations were utilized to verify the binding stability of monomeric components and protein targets. RESULTS: Macrophages had 140,690 gene targets, inflammatory response had 12,192 gene targets, intersection gene targets were 11,772. Key monomeric components (including: Pinocembrin, Fargesone-A, Nodakenin and Bowdichione) of LHQW were screened by HPLC. The results of cellular experiments indicated that LHQW could significantly reduce the mRNA expression of CCR5, CSF2, IFNG and TNF, thereby alleviating the inflammatory response caused by M1 macrophage. The computer simulations further validated the binding stability and conformation of key monomeric components and key protein targets, and IFNG/Nodakenin was able to form the most stable binding conformation for its action. CONCLUSION: In this study, the mechanism of LHQW inhibits the polarization of macrophages and the resulting inflammatory response was investigated by computer simulations and cellular experiments. We found that LHQW may not only reduce cell damage and death by acting on TNF and CCR5, but also inhibit the immune recognition process and inflammatory response by regulating CSF2 and IFNG to prevent polarization of macrophages. Therefore, these results suggested that LHQW may act through multiple targets to inhibit the polarization of macrophages and the resulting inflammatory response.
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Simulação por Computador , Medicamentos de Ervas Chinesas , Macrófagos , Humanos , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/química , Macrófagos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Inflamação , Anti-Inflamatórios/farmacologia , Células THP-1 , Biologia Computacional , Cromatografia Líquida de Alta PressãoRESUMO
The coexistence of wild boars and domestic pigs across Eurasia makes it feasible to conduct comparative genetic or genomic analyses for addressing how genetically different a domestic species is from its wild ancestor. To test whether there are differences in patterns of genetic variability between wild and domestic pigs at immunity-related genes and to detect outlier loci putatively under selection that may underlie differences in immune responses, here we analyzed 54 single-nucleotide polymorphisms (SNPs) of 19 immunity-related candidate genes on 11 autosomes in three pairs of wild boar and domestic pig populations from China, Iberian Peninsula, and Hungary. Our results showed no statistically significant differences in allele frequency and heterozygosity across SNPs between three pairs of wild and domestic populations. This observation was more likely due to the widespread and long-lasting gene flow between wild boars and domestic pigs across Eurasia. In addition, we detected eight coding SNPs from six genes as outliers being under selection consistently by three outlier tests (BayeScan2.1, FDIST2, and Arlequin3.5). Among four non-synonymous outlier SNPs, one from TLR4 gene was identified as being subject to positive (diversifying) selection and three each from CD36, IFNW1, and IL1B genes were suggested as under balancing selection. All of these four non-synonymous variants were predicted as being benign by PolyPhen-2. Our results were supported by other independent lines of evidence for positive selection or balancing selection acting on these four immune genes (CD36, IFNW1, IL1B, and TLR4). Our study showed an example applying a candidate gene approach to identify functionally important mutations (i.e., outlier loci) in wild and domestic pigs for subsequent functional experiments.
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Imunidade/genética , Polimorfismo de Nucleotídeo Único , Sus scrofa/genética , Suínos/genética , Animais , Antígenos CD36/genética , Frequência do Gene , Variação Genética , Genótipo , Interferons/genética , Interleucina-1beta/genética , Modelos Genéticos , Seleção Genética , Especificidade da Espécie , Receptor 4 Toll-Like/genéticaRESUMO
OBJECTIVE: To study the effects of silencing CLIC1 gene expression on the proliferation and invasion of Hca-F cells. METHODS: The mouse CLIC1 cDNA sequence was retrieved from NCBI. Three shRNA sequences were designed and cloned into pGPU6/GFP/Neo plasmids. The plasmids were transfected into Hca-F cells with Lipofectamine 2000. Cell Counting-8 (CCK-8) kit and transwell chamber were used to study the effects of CLIC1 on the proliferation and invasion of Hca-F cells. RESULTS: The pGPU6/GFP/Neo-shRNA-3 plasmid effectively repressed the expression of CLIC1 mRNA. Inhibition of CLIC1 gene expression led to decreased cell proliferation and reduced invasion. CONCLUSION: CLIC1 is essential for the proliferation and invasion of Hca-F cells.
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Carcinoma Hepatocelular/patologia , Proliferação de Células , Canais de Cloreto/genética , Neoplasias Hepáticas/patologia , Interferência de RNA , Animais , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Canais de Cloreto/antagonistas & inibidores , Canais de Cloreto/metabolismo , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Neoplasias Hepáticas/metabolismo , Camundongos , Invasividade Neoplásica , Plasmídeos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , TransfecçãoRESUMO
OBJECTIVE: To investigate the feasibility of monitoring therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on a rabbit VX2 liver tumor model by using phosphorous-31 magnetic resonance spectroscopy (31P MRS). METHODS: A total of 18 healthy New Zealand White rabbits were used to generate animal models by implanting VX2 tumor chips into livers through laparotomy. Tumor-bearing animals were randomly divided into three groups and were injected with AdCMVIL12-IRES-CKb, AdCMV-Empty and saline respectively via ear veins. 31P MRS scan was performed after animals were fed with creatine solution for five days. Animals were euthanized thereafter and tumors were removed for pathological examination, immunohistochemistry (IHC) staining and protein analysis (Western blot). RESULTS: The intrahepatic and seral expressions of creatine kinase (CKb) and IL-12 were detected only in AdCMVIL12-IRES-CKb group. Tumor diameters pre- and post- treatment in three groups were 1.63+/-0.04 vs 1.62+/-0.03 in AdCMVIL12-IRES-CKb group (P = 0.229), 1.59+/-0.05 vs 1.84+/-0.11 in AdCMV-Empty group (P = 0.003) and 1.60+/-0.02 vs 2.07+/-0.12 in saline group (P = 0.001), respectively. Pcr Changes between pre- and post- treatment among the three groups were compared (F = 6.235, P value is less than 0.05). PCr increased significantly in AdCMVIL12-IRES-CKb group as compared to AdCMV-Empty (P = 0.004) and saline group (P = 0.049), whereas no change found between AdCMV-Empty and saline group (P = 0.153). CONCLUSION: 31P MRS, an effective and non-invasive functional imaging method, can be used to monitor the therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on rabbit VX2 liver tumor model through detecting metabolic product of imaging reporter gene CKb (pCr).
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Creatina Quinase/metabolismo , Interleucina-12/genética , Neoplasias Hepáticas Experimentais/genética , Adenoviridae/genética , Animais , Creatina Quinase/genética , Expressão Gênica , Vetores Genéticos , Neoplasias Hepáticas Experimentais/patologia , Espectroscopia de Ressonância Magnética , CoelhosRESUMO
Lung cancer is the most common cause of cancer-related mortality worldwide. It is necessary to improve the understanding of the molecular mechanisms involved in lung cancer in order to develop more effective therapeutics for the treatment of this fatal disease. The canonical Wnt signaling pathway has been known to be important in a number of cancer types, including lung cancer. Pygopus (Pygo) is a recently identified downstream component of the Wnt signaling pathway required for ß-catenin/T-cell factor (TCF)-dependent transcription. However, the role of Pygo in lung cancer remains to be elucidated. The present study showed that Pygo2 is overexpressed in human lung cancer tissue samples and cell lines. Expression levels of Pygo2 were found to correlate with cytosolic ß-catenin protein levels in the samples examined. Co-immunofluorescent staining showed that Pygo2 protein accumulated in the nuclei and colocalized with nuclear ß-catenin in lung cancer cell lines expressing Pygo2. To investigate the functional importance of the Pygo2 overexpression in lung cancer, short hairpin RNA (shRNA) was used to knockdown Pygo2 mRNA in lung cancer cells expressing the gene. Pygo2 shRNA was observed to inhibit cell proliferation and decrease ß-catenin/TCF-dependent transcriptional activity in vitro. Furthermore, Pygo2 shRNA significantly suppressed lung cancer xenograft models in vivo (P<0.05). These results suggested that Pygo2 is a putative therapeutic target for human lung cancer and overexpression of Pygo2 may be important for aberrant Wnt activation in lung cancer.
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Trace constituents are widely present in complex mixtures, and trace analysis is challenging because of the unpredictable matrix. In this work, a high-component filtering strategy was developed for improved analysis of trace constituents in complex sample by liquid chromatography-mass spectrometry (LC-MS). Using a specifically designed chromatographic apparatus, the high-abundant fractions were filtered prior to LC-MS analysis. The samples complexity was reduced and the sample-loading amount for the rest low-level fractions can be considerably increased. The application of this approach was illustrated with an analytically challenging sample, a traditional Chinese herbal medicine named Compound Danshen Sample. We observed that the loss rate for 12 analytes during the filtering procedure ranged from 6.54 to 26.11%, but showed a stable repeatability with RSD<3.79%. The proposed filtering method with quadrupole time-of-flight mass spectrometritry (Q-TOF/MS) enhanced the detection capacity, offering a comprehensive characterization of 133 compounds in Compound Danshen Samples. The quantification sensitivity was also improved in trace analysis, allowing six low compounds that cannot be quantified by the traditional methods to be tested by the filtering method. It can be predicted that the qualitative and quantitative trace analysis will be greatly improved when the loading samples is increased resulting from the filtration of high-level targets. The proposed strategy is promising to monitor trace constituents in diverse complex mixtures in the analytical field of pharmaceutics, metabonomics and environments.
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Cromatografia Líquida , Misturas Complexas/análise , Medicamentos de Ervas Chinesas/análise , Filtração/métodos , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Misturas Complexas/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Plantas Medicinais , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
c-Jun N-terminal kinase (JNK) is located in focal adhesion plaque (FAP). JNK is necessary to growth, morphogenesis, and differentiation of cells; especially JNK1 has a close relation with tumors. In this study, we silenced JNK1 by using short hairpin RNA (shRNA) and examined the effect on migration and invasion of mouse hepatocellular carcinoma (HCC) cell line Hca-F in vitro. Three shRNA expression vectors (JNK1shRNA-1, JNK1shRNA-2, and JNK1shRNA-3) were constructed and transfected to Hca-F cells stably. The most effective shRNA was selected by detecting the expression levels of mRNA and protein. Transwell assay was performed to detect the ability of migration and invasion of cells. A negative control sequence (JNK1shRNA control) and non-transfected normal Hca-F cells were treated as control groups. The "Results" showed that the expression vectors of pSilencer-JNK1shRNA were constructed and transfected to Hca-F cells successfully. The most effective shRNA was JNK1shRNA-2. The expressions of mRNA and protein of JNK1 in Hca-F cells after transfection of JNK1shRNA-2 were decreased significantly compared with the other groups (all, P<0.01; all, P<0.05). The ability of migration and invasion was decreased after down-regulation of JNK1 expression (all, P<0.05). These results suggest that the inhibition of JNK1 expression can decrease ability of migration and invasion of mouse hepatocellular carcinoma cell line in vitro. JNK1 plays an important role in lymphatic metastasis of HCC. It may be a new target for gene therapy of lymphatic metastasis of HCC.