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1.
Clin Res Hepatol Gastroenterol ; 47(5): 102125, 2023 05.
Article in English | MEDLINE | ID: mdl-37062356

ABSTRACT

BACKGROUND: Traditional bismuth-containing quadruple therapy, as a first-line eradication treatment for Helicobacter pylori (H. pylori), has several disadvantages, including drug side effects, low medication adherence, and high costs. Trials of high-dose dual treatment have demonstrated its advantages, which include good safety and adherence profiles. In this study, we investigated the efficacy, safety, and compliance of a high-dose dual therapy when compared with bismuth-based quadruple treatment for the initial eradication of H. pylori infection on Hainan Island, China. METHODS: We randomized 846 H. pylori-infected patients into two groups. A bismuth-containing quadruple therapy group was administered the following: esomeprazole 20 mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily, and colloidal bismuth pectin in suspension 150 mg three times/day for 2 weeks. A high-dose dual therapy group was administered the following: esomeprazole 20 mg four times/day and amoxicillin 1000 mg three times/day for 2 weeks. Patients were given a 13C urea breath test at 4 weeks at treatment end. Adverse effects and compliance were evaluated at follow-up visits. RESULTS: Eradication rates in the high-dose dual therapy group were: 90.3% (381/422, 95% confidence interval [CI]: 87.1%-92.9%) in intention-to-treat (ITT) and 93.6% (381/407, 95% CI: 90.8%-95.8%) in per-protocol (PP) analyses. Eradication rates were 87.3% in ITT (370/424, 95% CI: 83.7%-90.3%) and 91.8% in PP analyses (370/403, 95% CI: 88.7%-94.3%) for quadruple therapy, with no statistical differences (P = 0.164 in ITT and P = 0.324 in PP analyses). Adverse effects were 13.5% (55/407) in the dual group and 17.4% (70/403) in the quadruple group (P = 0.129). Compliance was 92.4% (376/407) in the dual group and 86.6% (349/403) in the quadruple group (P = 0.007). CONCLUSIONS: High-dose dual therapy had high eradication rates comparable with bismuth-based quadruple treatment, with no differences in adverse effects, however higher adherence rates were recorded.


Subject(s)
Helicobacter Infections , Helicobacter pylori , Humans , Helicobacter Infections/drug therapy , Helicobacter Infections/etiology , Bismuth/therapeutic use , Bismuth/adverse effects , Anti-Bacterial Agents , Esomeprazole , Drug Therapy, Combination , Amoxicillin/adverse effects , Treatment Outcome , Proton Pump Inhibitors/adverse effects
2.
Mol Med Rep ; 23(2)2021 02.
Article in English | MEDLINE | ID: mdl-33313943

ABSTRACT

Pulmonary hypertension (PH) is a life­threatening disease that often involves vascular remodeling. Although pulmonary arterial smooth muscle cells (PASMCs) are the primary participants in vascular remodeling, their biological role is not entirely clear. The present study analyzed the role of enhancer of zeste homolog 2 (EZH2) in vascular remodeling of PH by investigating the behavior of PASMCs. The expression levels of EZH2 in PASMCs in chronic thromboembolic pulmonary hypertension (CTEPH), a type of PH, were detected. The role of EZH2 in PASMC migration was investigated by wound­healing assay following overexpression and knockdown. Functional enrichment analysis of the whole­genome expression profiles of PASMCs with EZH2 overexpression was performed using an mRNA Human Gene Expression Microarray. Quantitative (q)PCR was performed to confirm the results of the microarray. EZH2 expression levels increased in CTEPH cell models. The overexpression of EZH2 enhanced PASMC migration compared with control conditions. Functional enrichment analysis of the differentially expressed genes following EZH2 overexpression indicated a strong link between EZH2 and the immune inflammatory response and oxidoreductase activity in PASMCs. mRNA expression levels of superoxide dismutase 3 were verified by qPCR. The results suggested that EZH2 was involved in the migration of PASMCs in PH, and may serve as a potential target for the treatment of PH.


Subject(s)
Enhancer of Zeste Homolog 2 Protein/biosynthesis , Enhancer of Zeste Homolog 2 Protein/genetics , Hypertension, Pulmonary/genetics , Hypertension, Pulmonary/metabolism , Muscle, Smooth, Vascular/metabolism , Pulmonary Artery/metabolism , Cell Line , Cell Movement/genetics , Female , Gene Expression Regulation , Gene Knockdown Techniques , Gene Ontology , Humans , Hypertension, Pulmonary/chemically induced , Inflammation/genetics , Inflammation/immunology , Male , Middle Aged , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , NADPH Oxidase 1/metabolism , Oxidoreductases/metabolism , Superoxide Dismutase/metabolism , Tissue Array Analysis , Transcriptome , Vascular Remodeling/genetics
3.
Int J Nanomedicine ; 15: 2269-2276, 2020.
Article in English | MEDLINE | ID: mdl-32280219

ABSTRACT

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrotic interstitial pneumonia. And, oxidation/antioxidant imbalance plays an important role in the progress of IPF. Fullerene is considered to be a novel "structural" antioxidant. This study aimed to explore if water-soluble C60 (C60(OH)22) can exhibit antifibrotic activity in its antioxidant role. METHODS: Healthy C57BL/6J mice were randomly grouped and induced pulmonary fibrosis by intratracheal injection of bleomycin. RESULTS: The survival rate of mice was observed and found that 10mg/kg was the optimal dose of water-soluble C60 for pulmonary fibrosis. We observed that water-soluble C60 can alleviate the severity of pulmonary fibrosis by observing the chest computed tomography, pulmonary pathology, and content of collagen, alpha smooth muscle actin and fibronectin in lung. Compared with bleomycin group, ROS, the content of TNF-α in BALF, and the number of fibroblasts was significantly decreased and the number of type Ⅱ alveolar epithelial cells was increased after treatment with C60. CONCLUSION: Therefore, thanks to its powerful antioxidant action, water-soluble C60 can reduce the severity of pulmonary fibrosis induced by bleomycin in mice.


Subject(s)
Antioxidants/pharmacology , Bleomycin/adverse effects , Fullerenes/pharmacology , Idiopathic Pulmonary Fibrosis/drug therapy , Animals , Antioxidants/administration & dosage , Antioxidants/chemistry , Collagen , Disease Models, Animal , Fibroblasts/drug effects , Fibroblasts/pathology , Fullerenes/administration & dosage , Fullerenes/chemistry , Idiopathic Pulmonary Fibrosis/chemically induced , Idiopathic Pulmonary Fibrosis/pathology , Lung/drug effects , Lung/metabolism , Lung/pathology , Male , Mice, Inbred C57BL , Solubility , Water/chemistry
4.
J Cell Biochem ; 121(4): 2938-2949, 2020 04.
Article in English | MEDLINE | ID: mdl-31692072

ABSTRACT

BACKGROUND: Calcium-binding tyrosine phosphorylation-regulated protein (CABYR) is a group of isoforms produced by alternative splicing and is overexpressed in human malignancies including hepatocellular carcinoma (HCC). However, the prognostic value and biological functions of its major protein isoforms, named CABYR-a/b (combined CABYR-a and CABYR-b), in HCC remain to be established. METHODS: CABYR-a/b expression was detected in HCC tissues and cell lines by quantitative real-time polymerase chain reaction and Western blot analysis. The correlation of CABYR-a/b expression with clinical characteristics and its prognosis impact were determined by statistical analysis. Finally, the biological functions and molecular mechanism of CABYR-a/b were also investigated using molecular biology approaches. RESULTS: The present research found that CABYR-a/b was markedly elevated in HCC specimens and cell lines. Upregulated CABYR-a/b level had positive association with tumor size and differentiation in patients. Moreover, cases with elevated CABYR-a/b level had poorer overall survival (OS) and disease-free survival (DFS) than those with reduced CABYR-a/b level. Multivariate analysis and prognostic nomograms demonstrated that CABYR-a/b overexpression was an independent predictive indicator for OS and DFS. The calibration curve for the odds of OS and DFS demonstrated that the prediction by nomograms was in excellent accordance with actual situation. CABYR-a/b downregulation suppressed cell proliferation and induced G1-phase arrest via decreasing cyclin D1 and cyclin dependent kinase 4, while promoted apoptosis by reducing B-cell lymphoma 2 (Bcl-2) and increasing Bcl-2-associated death promoter. CONCLUSION: Our research indicates that CABYR-a/b exerts an oncogenic effect on HCC development and may become a new prognostic indicator for patients with HCC.


Subject(s)
Apoptosis , Calcium-Binding Proteins , Calcium/metabolism , Carcinoma, Hepatocellular/metabolism , Liver Neoplasms/metabolism , Tyrosine/chemistry , Aged , Alternative Splicing , Biomarkers, Tumor/metabolism , Calcium-Binding Proteins/metabolism , Carcinoma, Hepatocellular/diagnosis , Cell Differentiation , Cell Line, Tumor , Cell Proliferation , Cyclin D1/metabolism , Cyclin-Dependent Kinase 4/metabolism , Disease-Free Survival , Female , Gene Expression Regulation, Neoplastic , Humans , Liver Neoplasms/diagnosis , Male , Middle Aged , Phosphorylation , Prognosis , Protein Binding , Protein Isoforms , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Small Interfering/metabolism , Treatment Outcome
5.
Biomed Pharmacother ; 112: 108611, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30797148

ABSTRACT

Circular RNAs (circRNAs) are a novel class of non-coding RNAs with distinct properties and diverse physiological and pathological functions. However, the functions of circRNAs in colorectal cancer (CRC) remain elusive. This study aimed to investigate the functional roles of circVAPA in CRC. High-throughput RNA sequencing was performed in 4 paired CRC tissues, and circVAPA (hsa_circ_0006990), was identified as a potential functional circRNA. Using quantitative real-time polymerase chain reaction (qRT-PCR), circVAPA was found to be up-regulated in CRC patients' tissues and plasma. Furthermore, circVAPA level was associated with unfavorable clinicopathologic features in CRC. The area under curve (AUC) of ROC was 0.724, suggesting that plasma level of circVAPA could serve as a promising biomarker for CRC detection. Sanger sequencing confirmed the back-splice junction sequences of circVAPA. Actinomycin D and RNase R treatments suggested that circVAPA was highly stable compared with its linear counterpart, and qRT-PCR for the circVAPA level in nuclear and cytoplasmic fractions indicated that circVAPA was predominantly localized in the cytoplasm. Gain-of-function and loss-of-function studies in CRC cell lines indicated that circVAPA could promote CRC cell proliferation, migration, invasion, and inhibit apoptosis. miRanda software (v3.3a) was used to predict target miRNAs of circVAPA. Moreover, target miRNAs associated with the KEGG pathway of COLORECTAL CANCER (Entry: map05210; https://www.kegg.jp/) were screened using DIANA-miRPath v.3 platform (Reverse Search module; TarBase v7.0 method). The analyses by miRanda and miRPath suggested that circVAPA could potentially bind to hsa-miR-101-3p (miR-101) associated with the COLORECTAL CANCER pathway. Luciferase reporter assay confirmed a direct interaction between circVAPA and miR-101. Furthermore, circVAPA had no effect on the expression level of miR-101, and miR-101 over-expression had the similar tumor-suppressing effects as circVAPA silencing. The tumor-promoting effect of circVAPA over-expression could be reversed by the up-regulation of miR-101. These data demonstrated that circVAPA promoted CRC progression by sponging miR-101. In conclusion, we have verified that circVAPA is up-regulated in CRC patients' tissues and plasma, and exerts oncogenic properties by sponging miR-101 in CRC. CircVAPA could serve as a promising biomarker and a therapeutic target for CRC.


Subject(s)
Biomarkers, Tumor/biosynthesis , Colorectal Neoplasms/metabolism , MicroRNAs/biosynthesis , RNA/biosynthesis , Up-Regulation/physiology , Vesicular Transport Proteins/biosynthesis , Aged , Biomarkers, Tumor/genetics , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Female , HCT116 Cells , HEK293 Cells , HT29 Cells , Humans , Male , MicroRNAs/genetics , Middle Aged , RNA/genetics , RNA, Circular , Vesicular Transport Proteins/genetics
6.
Biomed Pharmacother ; 105: 1147-1154, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30021351

ABSTRACT

microRNA-19a-3p (miR-19a-3p) has been reported to regulate cell proliferation in hepatocellular carcinoma (HCC), but its role in HCC metastasis remains unknown. In this study, miR-19a-3p was noted to be upregulated in HCC specimens and cell lines. Aberrant expression of miR-19a-3p stimulated HCC cell metastasis, and phosphatase and tensin homolog (PTEN) was shown to be a direct target of miR-19a-3p. miR-19a-3p-mediated HCC metastasis was reversed by restoration of PTEN or could be imitated by silencing of PTEN. Modulation of miR-19a-3p also altered expression of phosphorylated Akt, a downstream mediator of PTEN. Moreover, aberrant expression of miR-19a-3p induced sorafenib resistance by regulating the PTEN/Akt pathway. In conclusion, ectopic expression of miR-19a-3p contributes to HCC metastasis and chemoresistance by modulating PTEN expression and the PTEN-dependent pathways.


Subject(s)
Carcinoma, Hepatocellular/metabolism , Drug Resistance, Neoplasm/physiology , Liver Neoplasms/metabolism , MicroRNAs/biosynthesis , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Hepatocytes/metabolism , Hepatocytes/pathology , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , MicroRNAs/genetics , Signal Transduction/physiology
7.
Am J Physiol Lung Cell Mol Physiol ; 314(2): L276-L286, 2018 02 01.
Article in English | MEDLINE | ID: mdl-29122755

ABSTRACT

Pulmonary arterial smooth muscle cell (PASMC) migration plays a key role in vascular remodeling, which occurs during development of chronic thromboembolic pulmonary hypertension (CTEPH). Activation of the renin-angiotensin system (RAS) contributes to vascular remodeling observed in many diseases, including idiopathic pulmonary arterial hypertension. However, the role of RAS imbalance in CTEPH has not been characterized. Here, we hypothesize that RAS imbalance regulates vascular remodeling by promoting PASMC migration in CTEPH. Serum renin and angiotensin II levels in patients with CTEPH were quantified by ELISA. The pulmonary endarterectomy tissues were stained and analyzed by immunohistochemistry. PASMCs were isolated and verified by immunofluorescence staining. PASMC migration was determined by Transwell assay. Phosphorylation and protein level were detected by Western blotting. Serum levels of renin and angiotensin II were increased in patients with CTEPH {renin [median (25th percentile, 75th percentile) in pg/ml], 1,199.94 [690.85, 1,656.90] vs. 595.43 [351.48, 936.43], P < 0.001; angiotensin II [in pg/ml], 63.97 [45.97, 345.24] vs. 56.85 [11.20, 90.37], P < 0.05}. The migration of PASMCs isolated from patients with CTEPH was enhanced compared with control. Angiotensin II promoted the migration of PASMCs via activation of angiotensin II receptor 1 and phosphorylation of ERK1/2, whereas angiotensin-(1-7) counteracted this effect through activation of the Mas receptor and ERK1/2. These results demonstrate that the renin-angiotensin system regulates migration of PASMCs from patients with CTEPH via the ERK1/2 pathway. Our findings suggest that angiotensin-(1-7) or reagents targeting the renin-angiotensin system will be beneficial in the development of novel therapies for CTEPH.


Subject(s)
Cell Movement , Hypertension, Pulmonary/pathology , Muscle, Smooth, Vascular/pathology , Pulmonary Artery/pathology , Thromboembolism/pathology , Angiotensin II/metabolism , Cell Proliferation , Cells, Cultured , Female , Humans , Hypertension, Pulmonary/metabolism , MAP Kinase Signaling System , Male , Middle Aged , Mitogen-Activated Protein Kinases , Muscle, Smooth, Vascular/metabolism , Phosphorylation , Pulmonary Artery/metabolism , Renin/metabolism , Renin-Angiotensin System , Signal Transduction , Thromboembolism/metabolism
8.
Oncol Lett ; 14(3): 2703-2708, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28928813

ABSTRACT

The overexpression of heat shock protein 70 (HSP70), a major stress-inducible heat shock protein, has been identified to enhance the proliferation, survival, invasion and metastasis of diverse types of human cancer. However, its role in hepatocellular carcinoma (HCC) remains poorly understood. The present study demonstrated that HSP70 expression was higher in tested HCC cell lines, compared with the normal hepatocyte LO2, and the suppression of HSP70 significantly inhibited the proliferation of SMMC-7721 and Hep3B cells. The growth inhibitory effect was mediated by cell cycle arrest at the G1/S phase with reduced cyclin D1 and increased p27Kip1 expression. Furthermore, HSP70 knockdown significantly inhibited the migration and invasion abilities of HCC cells. In conclusion, HSP70 is a key regulator involved in the proliferation, migration and invasion of HCC, and it may be used as a potential therapeutic target for HCC.

9.
J Cancer Res Clin Oncol ; 142(11): 2309-17, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27601163

ABSTRACT

PURPOSE: Aberrant expression of eukaryotic initiation factor 4E (eIF4E) has been observed in human malignancies. However, its role in hepatocellular carcinoma (HCC) remains to be established. The purpose of this study was to detect eIF4E expression and to evaluate its clinical relevance. METHODS: The eIF4E expression was studied in ninety HCC and randomly selected thirty-one non-tumor tissues from the same patient cohort, as well as in normal hepatic and HCC cell lines. The relation between its expression and clinicopathological parameters was also analyzed. RESULTS: eIF4E expression was higher in HCC samples and cell lines compared with that in non-tumor tissues (P < 0.001) and hepatocyte LO2, respectively. eIF4E overexpression was significantly associated with tumor number (P = 0.005) and incomplete encapsulation (P = 0.001). The 5-year overall survival rate and disease-free survival rate for patients with high eIF4E expression were 32.5 and 31.2 %, respectively; and for low eIF4E expression, it was 67.9 and 64.4 %, respectively (P < 0.001). Furthermore, subgroup analysis showed that high eIF4E level predicted poorer overall survival only for incomplete encapsulation (P = 0.001) and cirrhosis (P < 0.001), but not for complete encapsulation (P = 0.804) and non-cirrhosis (P = 0.359). Multivariate analysis revealed that eIF4E overexpression was an independent indicator for both overall survival (hazard ratio, 2.015; P = 0.043) and disease-free survival (hazard ratio, 2.666; P = 0.006). CONCLUSIONS: eIF4E protein might result in the malignant progression of HCC, and its overexpression may be a powerful prognostic biomarker and therapeutic target for HCC patients.


Subject(s)
Carcinoma, Hepatocellular/metabolism , Liver Neoplasms/metabolism , Nucleocytoplasmic Transport Proteins/metabolism , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Disease Progression , Female , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Liver Neoplasms/pathology , Male , Middle Aged , Neoplasm Recurrence, Local/metabolism , Neoplasm Recurrence, Local/pathology , Nucleocytoplasmic Transport Proteins/biosynthesis , Prognosis , Survival Rate , Tissue Array Analysis , Up-Regulation
10.
Oncol Rep ; 36(3): 1483-90, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27460949

ABSTRACT

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. Src is involved in multiple processes of cancer metastasis; however, its significance in HCC is not well defined. In the present study, overexpression of Src phosphorylation (Y416) was observed in the highly metastatic MHCC97H cell line; additionally, through inhibition of Src kinase activation, HCC cell proliferation, migration, invasion and colony formation were significantly reduced in vitro. Tumour growth was not affected in the orthotopic xenograft HCC model, but the metastasic potential was inhibited as revealed by reduced lung metastasic foci after administration of saracatinib. Phosphorylation level of Src pathway signalling molecules, such as Src, FAK and Stat3, were also reduced in vitro and in vivo, as a result of the anti-metastasic effects caused by saracatinib treatment. In conclusion, we demonstrated the pro-metastasic role of Src in HCC, and further experiments suggest the use of the Src inhibitor in combination with cytotoxic agents and other anticancer treatments to improve HCC prognosis.


Subject(s)
Benzodioxoles/pharmacology , Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , Lung Neoplasms/drug therapy , Neoplasm Metastasis/drug therapy , Quinazolines/pharmacology , Animals , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Humans , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness/pathology , Neoplasm Metastasis/pathology , Phosphorylation/drug effects , Signal Transduction/drug effects , Xenograft Model Antitumor Assays/methods , src-Family Kinases/metabolism
11.
Talanta ; 147: 207-12, 2016 Jan 15.
Article in English | MEDLINE | ID: mdl-26592597

ABSTRACT

We report a simple one-step hydrothermal method for the synthesis of hydrophilic luminescent upconversion nanoparticles (UCNPs) using malonic acid as the stabilizer and functional agent. Using this method, two UCNPs with different colors of upconversion luminescence were synthesized. The surface of the as-prepared UCNPs was capped with carboxyl groups, which not only resulted in the UCNPs having good dispersity in water, but also allowed further conjugation with other functional molecules, thus indicating the potential applications in biosensing and bioimaging. To demonstrate this, amino-labeled single-stranded DNA (ssDNA) was conjugated on the surface of the UCNPs. Based on the different absorption and luminescence quenching abilities of graphene oxide (GO) to ssDNA-modified UCNPs before and after exonuclease I (Exo I)-triggered hydrolysis of ssDNA, a detection platform was developed for the detection of Exo I activity with a detection limit of 0.02U mL(-1). The prepared hydrophilic UCNPs were also used successfully for in vivo upconversion luminescence imaging of nude mice.


Subject(s)
Biosensing Techniques/methods , Luminescent Agents/chemistry , Nanoparticles/chemistry , Optical Imaging/methods , Animals , Chemistry Techniques, Synthetic , DNA, Single-Stranded/metabolism , Exodeoxyribonucleases/metabolism , Graphite/chemistry , Hydrolysis , Hydrophobic and Hydrophilic Interactions , Mice , Molecular Imaging , Oxides/chemistry
12.
Nucleic Acids Res ; 42(13): 8719-31, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24939896

ABSTRACT

Ligands that can interact specifically with telomeric multimeric G-quadruplexes could be developed as promising anticancer drugs with few side effects related to other G-quadruplex-forming regions. In this paper, a new cationic porphyrin derivative, m-TMPipEOPP, was synthesized and characterized. Its multimeric G-quadruplex recognition specificity under molecular crowding conditions was compared to its isomer p-TMPipEOPP. The slight structural difference accounts for different multimeric G-quadruplex recognition specificity for the two isomers. p-TMPipEOPP can barely discriminate between multimeric and monomeric G-quadruplexes. By contrast, m-TMPipEOPP can bind with multimeric but not with monomeric G-quadruplexes. p-TMPipEOPP might bind to multimeric G-quadruplexes by two modes: sandwich-like end-stacking mode and pocket-dependent intercalative mode. Increasing the pocket size between adjacent two G-quadruplex units is beneficial for the latter mode. m-TMPipEOPP might bind to multimeric G-quadruplexes by a side binding mode, which confers m-TMPipEOPP with higher multimeric G-quadruplex recognition specificity compared to p-TMPipEOPP. m-TMPipEOPP increases the stability of multimeric G-quadruplex under both dilute and molecular crowding conditions but its G-quadruplex-stabilizing ability is a little weaker than p-TMPipEOPP. These results provide important information for the design of highly specific multimeric G-quadruplex ligands. Another interesting finding is that pocket size is an important factor in determining the stability of multimeric G-quadruplexes.


Subject(s)
G-Quadruplexes , Porphyrins/chemistry , DNA/chemistry , DNA/metabolism , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/metabolism , Fluorescence , Isomerism , Piperidines
13.
PLoS One ; 8(9): e73012, 2013.
Article in English | MEDLINE | ID: mdl-24039849

ABSTRACT

Label-free metal ion detection methods were developed. To achieve these, a reconstructed Cu(2+)-specific DNA-cleaving DNAzyme (Cu(2+)-specific DNAzyme) with an intramolecular stem-loop structure was used. G-quadruplex-forming G-rich sequence(s), linked at the ends of double-helix stem of an intramolecular stem-loop structure, was partly caged in an intramolecular duplex or formed a split G-quadruplex. Cu(2+)-triggered DNA cleavage at a specific site decreased the stability of the double-helix stem, resulting in the formation or destruction of G-quadruplex DNAzyme that can effectively catalyze the 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS)-H2O2 reaction. Based on these, two label-free, cost-effective and simple Cu(2+) sensors were designed. These two sensors followed different detection modes: 'turn-on' and 'turn-off'. As for the 'turn-on' sensor, the intramolecular stem-loop structure ensured a low background signal, and the co-amplification of detection signal by dual DNAzymes (Cu(2+)-specific DNAzyme and G-quadruplex DNAzyme) provided a high sensitivity. This sensor enabled the selective detection of aqueous Cu(2+) with a detection limit of 3.9 nM. Visual detection was possible. Although the 'turn-off' sensor gave lower detection sensitivity than the 'turn-on' one, the characteristics of cost-effectiveness and ease of operation made it an important implement to reduce the possibility of pseudo-positive or pseudo-negative results. Combining the ability of Hg(2+) ion to stabilize T-T base mismatch, above dual DNAzymes-based strategy was further used for Hg(2+) sensor design. The proposed sensor allowed the specific detection of Hg(2+) ion with a detection of 4.8 nM. Visual detection was also possible.


Subject(s)
Biosensing Techniques/methods , Copper/chemistry , DNA, Catalytic/chemistry , G-Quadruplexes , Mercury/chemistry , Colorimetry/methods , Sensitivity and Specificity , Spectrophotometry, Ultraviolet , Staining and Labeling
14.
Biosens Bioelectron ; 42: 225-8, 2013 Apr 15.
Article in English | MEDLINE | ID: mdl-23208090

ABSTRACT

A previously reported Cu²âº-dependent DNAzyme/substrate complex was reconstructed in this work, which makes possible the use of an intramolecular stem-loop structure and is, therefore, a good choice for the design of Cu²âº sensors. To demonstrate this, a fluorescent sensor was designed on the basis of the reconstructed complex. In this sensor, the fluorophore/quencher pair was caged tightly in an intramolecular double-helix structure; thus, the background signal was greatly suppressed. Cu²âº-dependent cleavage of the complex could cause the release of the fluorophore, leading to restoration of the fluorescence signal. High quenching efficiency provides the sensor with three important characteristics: high sensitivity, high temperature variation tolerance and high ionic strength tolerance. The proposed sensor allows specific detection of aqueous Cu²âº down to a limit of 0.6 nM, and the performance is independent of temperature and ionic strength in the range of 4-40 °C and 0.8-3.0 M NaCl, respectively. This work identifies a good choice for sensor design on the basis of DNAzymes containing triple-helix structures.


Subject(s)
Biosensing Techniques/methods , Copper/isolation & purification , DNA, Catalytic/chemistry , Copper/chemistry , Fluorescence , Osmolar Concentration , Substrate Specificity
15.
Asian Pac J Cancer Prev ; 13(12): 6455-61, 2012.
Article in English | MEDLINE | ID: mdl-23464474

ABSTRACT

OBJECTIVE: E-cadherin has been identified as a tumor suppressor in many types of carcinoma. However, some studies recently suggested that the role and expression of E-cadherin might be more complex and diverse. In the present study, we evaluated the prognostic value of E-cadherin expression with reference to levels in membranes and cytoplasm, and the membrane/cytoplasm ratio, in hepatocellular carcinomas (HCCs) after curative hepatectomy. METHODS: The expression of E-cadherin was assessed by immunohistochemistry in HCC tissue microarrays from 125 patients, and its prognostic values and other clinicopathlogical data were retrospectively analyzed. Patients were followed for a median period of 43.7 months (range 1 to 126 months). RESULTS: Univariate analysis demonstrated that a high membrane/cytoplasm (M/C) ratio of E-cadherin expression was associated with poor overall survival (OS) (P =0.001) and shorter time to recurrence (TTR) (P =0.038), as well as tumor size, intrahepatic metastasis, and TNM stage. In contrast, neither membrane nor cytoplasmic expression of E-cadherin was related with OS and TTR. Furthermore, multivariate analysis confirmed the M/C ratio to be an independent predictor of OS (P =0.031). ?2 tests additionally showed that the M/C ratio of E-cadherin expression was related with early stage recurrence (P =0.012), rather than later stage recurrence. CONCLUSION: The M/C ratio of E-cadherin expression is a strong predictor of postoperative survival and is associated with early stage recurrence in patients with HCC.


Subject(s)
Cadherins/genetics , Cadherins/metabolism , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/metabolism , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Adolescent , Adult , Aged , Carcinoma, Hepatocellular/pathology , Cell Membrane/genetics , Cell Membrane/metabolism , Cytoplasm/genetics , Cytoplasm/metabolism , Female , Hepatectomy/methods , Humans , Liver Neoplasms/pathology , Male , Middle Aged , Neoplasm Staging/methods , Prognosis , Retrospective Studies , Young Adult
16.
Parasit Vectors ; 4: 168, 2011 Aug 28.
Article in English | MEDLINE | ID: mdl-21871123

ABSTRACT

BACKGROUND: Toxoplasma gondii has been shown to trigger strong cellular immune responses to heterologous antigens expressed by the parasite in the inbred mouse model. We studied the immune response induced by T. gondii as an effective vaccine vector in chickens and rabbits. RESULTS: T. gondii RH strain was engineered to express the yellow fluorescent protein (YFP) in the cytoplasm. A subcutaneous injection of the transgenic T. gondii YFP in chickens afforded partial protection against the infection of transgenic E. tenella YFP. T. gondii YFP induced low levels of antibodies to YFP in chickens, suggesting that YFP specific cellular immune response was probably responsible for the protective immunity against E. tenella YFP infection. The measurement of T-cell response and IFN-γ production further confirmed that YFP specific Th1 mediated immune response was induced by T. gondii YFP in immunized chickens. The transgenic T. gondii stimulated significantly higher YFP specific IgG titers in rabbits than in chickens, suggesting greater immunogenicity in a T. gondii susceptible species than in a resistant species. Priming with T. gondii YFP and boosting with the recombinant YFP can induce a strong anti-YFP antibody response in both animal species. CONCLUSIONS: Our findings suggest that T. gondii can be used as an effective vaccine vector and future research should focus on exploring avirulent no cyst-forming strains of T. gondii as a live vaccine vector in animals.


Subject(s)
Drug Carriers , Genetic Vectors , Protozoan Vaccines/immunology , Toxoplasma/immunology , Animals , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Chickens , Immunoglobulin G/blood , Injections, Subcutaneous , Luminescent Proteins/genetics , Luminescent Proteins/immunology , Protozoan Vaccines/administration & dosage , Rabbits , Recombinant Proteins/genetics , Recombinant Proteins/immunology , T-Lymphocytes/immunology , Toxoplasma/genetics , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/immunology
17.
Zhonghua Nei Ke Za Zhi ; 50(5): 370-3, 2011 May.
Article in Chinese | MEDLINE | ID: mdl-21624216

ABSTRACT

OBJECTIVE: To evaluate the effect of enteral nutrition (EN) versus total parenteral nutrition (TPN) on gut barrier function in patients with severe acute pancreatitis (SAP). METHODS: Sixty-three patients with SAP enrolled from 4 hospitals were randomly assigned into EN group (29 cases) and TPN group (34 cases). EN group patients were fed via a spiral nasojejunal feeding tube placed routinely by endoscopy or fluoroscopy, and TPN group patients were nourished intravenously with TPN during the same period. The changes of serum endotoxin, diamine oxidase, and urinary excretion of lactulose and mannitol ratio (L/M) were observed. RESULTS: Plasma concentration of endotoxin were markedly decreased in EN group as compared with that in TPN group at the 7(th), 14(th), 21(th) day of entry trial [(39.30 ± 15.82) EU/L vs (73.05 ± 21.16) EU/L, (22.64 ± 14.31) EU/L vs (49.34 ± 24.54) EU/L, (14.81 ± 10.93) EU/L vs (30.08 ± 14.10) EU/L, P < 0.05]. Plasma concentration of diamine oxidase were markedly decreased in EN group as compared with that in TPN group at the 7(th), 14(th) day of entry trial [(9.97 ± 3.84) U/L vs (19.89 ± 9.89) U/L, (5.42 ± 1.84) U/L vs (8.79 ± 4.08) U/L, both P < 0.05]. The urinary L/M decreased significantly in EN group than those in TPN group at the 7(th), 14(th), 21(th) day of entry trial (0.28 ± 0.25 vs 0.65 ± 0.45, 0.21 ± 0.18 vs 0.54 ± 0.41, 0.08 ± 0.04 vs 0.29 ± 0.06, all P < 0.05). CONCLUSION: EN has better effect on improving intestinal barrier function than TPN in treatment of patients with SAP.


Subject(s)
Enteral Nutrition , Pancreatitis, Acute Necrotizing/therapy , Parenteral Nutrition, Total , Adult , Aged , Female , Gastrointestinal Tract , Humans , Male , Middle Aged , Pancreatitis, Acute Necrotizing/physiopathology , Treatment Outcome , Young Adult
18.
Zhonghua Yi Xue Za Zhi ; 88(34): 2407-9, 2008 Sep 09.
Article in Chinese | MEDLINE | ID: mdl-19087716

ABSTRACT

OBJECTIVE: To investigate the effects of continuous early enteral nutrition (EEN) supplemented with glutamine and arginine on gut barrier function in patients with severe acute pancreatitis (SAP). METHODS: Thirty two patients with a diagnosis of acute pancreatitis predicted to develop severe disease were randomized into 2 groups: EEN group (n = 18) and EEN + glutamine and arginine group (enteral immunonutrition group, n = 14). EEN was initiated when homeostasis was achieved within 72 hours after attack, and both group received isocaloric isonitrogenous nutrition. Glutamine and arginine were administered into jejunum in the enteral immunonutrition group. Serum amylase, plasma diamine oxidase (DAO), C-reactive protein (CRP), plasma endotoxin, urinary excretion of lactulose (L), and mannitol (M) were measured, and APACHE-II scores were recorded on days 1, 7, and 14. Complications, and length and cost of hospitalization were recorded as well. RESULTS: EEN and enteral immunonutrition were both tolerated well. There was no difference in APACHE-IIscore between the two groups (P > 0.05). The DAO, CRP, plasma endotoxin, and urinary L/M levels decreased with the course of SAP. However, the plasma endotoxin and urinary L/M on day 7 of the enteral immunonutrition group were (10.0 +/- 3.8) EU/ml and 0.29 +/- 0.15 respectively, both significantly higher than those of the EEN group [(7.9 +/- 2.8) EU/ml and 0.16 +/- 0.08 respectively, both P < 0.05]. The length of hospital stay and cost showed no differences between the two groups. CONCLUSION: EEN is safe and feasible in treatment of SAP. Enteral immunonutrition containing glutamine and arginine improves the gut barrier function by reducing the gut permeability and decreasing plasma endotoxin level in the early stage of SAP.


Subject(s)
Enteral Nutrition , Pancreatitis, Acute Necrotizing/physiopathology , Pancreatitis, Acute Necrotizing/therapy , Adult , Arginine/administration & dosage , Female , Gastrointestinal Tract , Glutamine/administration & dosage , Humans , Male , Middle Aged , Prospective Studies
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