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Objective:To investigate the effect and mechanism of Yiqi Fuzheng Jiedu decoction （YQFZJDD） on autophagy and growth of A549 cells. Method:A549 cells were intervened with YQFZJDD-containing serum prepared in advance. The levels of microtubule-associated protein 1 light chain 3 （LC3）， homologue of yeast autophagy-related gene 6 （Beclin1）， p62， p53， adenosine 5'-monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mammalian target of rapamycin （mTOR）， and phosphorylated mTOR （p-mTOR） were detected by Western blot assay， and microtubule-associated protein 1A/1B light chain 3B （MAP1LC3B） by immunofluorescence （IF） assay. The proliferation， invasion， and senescence of A549 cells were separately measured by 5-ethynyl-2'-deoxyuridine （EDU） staining， Transwell assay， and <italic>β</italic>-galactosidase staining. In the presence of autophagy inhibitor 3-methyladenine （3-MA， 5 mmol·L<sup>-1</sup>）， the cells were divided into the 10% fetal bovine serum （blank） group， 10% control serum （control） group， low-， medium-， and high-dose 10% YQFZJDD-containing serum groups， and high-dose 10% YQFZJDD-containing serum + 3-MA group， followed by the measurement of A549 cell proliferation， invasion， and senescence. In the adoption of p53 inhibitor Pifithrin-<italic>α</italic> （PFT-<italic>α</italic>， 10 μmol·L<sup>-1</sup>）， the cells were divided into the control group， PFT-<italic>α</italic> group， high-dose YQFZJDD-containing serum group， and high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group. Then the LC3-Ⅱ and Beclin1 expression， MAP1LC3B fluorescence intensity， as well as A549 cell proliferation， invasion and senescence were determined. Result:Compared with the blank group and control group， YQFZJDD-containing serum at the medium and high doses up-regulated the protein expression levels of LC3-Ⅱ and Beclin1 in A549 cells after 48-h intervention in a dose-dependent manner （<italic>P</italic><0.01）. Besides， YQFZJDD-containing serum at the low-， medium-， and high-doses down-regulated p62 and p-mTOR/mTOR expression （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated p53 and p-AMPK/AMPK （<italic>P</italic><0.01）， decreased the number of proliferative and invasive cells， increased the number of senescent cells （<italic>P</italic><0.01）， and enhanced the IF intensity of MAP1LC3B， with the optimal effect observed in the high-dose YQFZJDD-containing serum group （<italic>P</italic><0.01）. Compared with the high-dose YQFZJDD-containing serum group， the high-dose YQFZJDD-containing serum + 3-MA group and high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group exhibited significantly increased proliferative and invasive cells but decreased senescent cells （<italic>P</italic><0.05， <italic>P<</italic>0.01）. Meanwhile， the IF intensity of MAP1LC3B in the high-dose YQFZJDD-containing serum + PFT-<italic>α</italic> group was weakened and the LC3-Ⅱ and Beclin1 protein expression levels declined （<italic>P</italic><0.05， <italic>P<</italic>0.01）. Conclusion:YQFZJDD promotes the autophagy of A549 cells through the p53/AMPK signaling pathway to inhibit their proliferation， invasion and migration but accelerate senescence， thus playing a crucial role in inhibiting the progression of lung cancer.
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Objective:To investigate the effect of modified Ditantang on autophagy and relevant proteins in brain cells of rats with cerebral ischemia reperfusion injury. Method:The cerebral ischemic reperfusion (CIR) injury model in rats was built by reversible middle cerebral artery occlusion artery suture of middle cerebral embolism method, and randomly divided into sham group, model group, high-dose modified Ditantang group(H-dose), low-dose modified Ditantang group (L-dose, 0.384 g·kg-1) and PLXT group (0.1 g·kg-1). Sham and model groups were given normal saline by gastric perfusion, H-dose and L-dose groups were given modified Ditantang, and the PLXT group were given Piracetam tablets, intragastric volume 10 mL·kg-1. The treatment lasted for 7 d. Within 24 hours after administration, the histopathological examination was performed, the volume of cerebral infarction, neuronal apoptosis and serum levels of inflammatory factors were compared, and the expressions of autophagy related microtuble-associated protein 1 light chain 3(LC3)Ⅱ, Beclin1, B-cell lymphoma-2(Bcl-2) and p62 in brain tissues were determined. Result:Cells and blood vessel necrosis, neuron swelling and interstitial edema were observed in model group, a few neurons died, edema was reduced, swelling of nerve cells was alleviated in H-dose, L-dose and PLXT groups. The volume of cerebral infarction and neuronal apoptosis in H-dose, L-dose and PLXT groups were lower than those in model group (Pα, intedeukin (IL)-2 and IL-8 in H-dose, L-dose and PLXT groups were lower than those in model group (PPConclusion:Modified Ditantang can improve brain injury and interfere with autophagy after MCAO/R, alleviate inflammation, and regulate autophagic activity, which may be related to the down-regulation of expressions of LC3 Ⅱ, Beclin1 and the up-regulation of expression of p62.
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AIM: To investigate the synergistic antitumor effect of sinomenine(SIN)and cisplatin(DDP) against human fibrosarcoma HT-1080 cells and its possible mechanism.METHODS:CCK-8 assay was used to detect the cytotoxic effect of SIN and DDP in HT-1080 cells,and the combination index(CI)of SIN and DDP was analyzed by Chou-Talalay method.The apoptosis was analyzed by flow cytometry.The protein levels of copper transporter 1(CTR1),gluta-thione S-transferase-π(GST-π),Bcl-2 and Bax were determined by Western blot.RESULTS: At 48 h after treatment with SIN or DDP at various concentrations,the cell viability was remarkably reduced(P<0.05), and the IC50values of DDP and SIN were 6.50 μmol/L and 1.06 mmol/L, respectively.When the inhibition rate of HT-1080 cells exceeded 25%, SIN in combination with DDP produced a synergistic effect(CI<1).Combination treatment of SIN with DDP signi-ficantly induced apoptosis of HT-1080 cells.SIN up-regulated the protein levels of CTR1 and Bax,and down-regulated the protein levels of GST-πand Bcl-2(P<0.05).CONCLUSION:SIN synergistic induces apoptosis with DDP in human fi-brosarcoma HT-1080 cells by up-regulating the protein levels of CTR1 and Bax, and down-regulating the protein levels of GST-πand Bcl-2.
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<p><b>OBJECTIVE</b>To evaluate the value of echocardiography in the diagnosis of noncompaction of the ventricular myocardium (NCVM) and to elucidate the echocardiographic characteristics of NCVM.</p><p><b>METHODS</b>This study included 53 patients (28 boys and 25 girls), with an age for initial diagnosis of 15 days to 18 years, who were diagnosed with NCVM by echocardiography between May 2006 and May 2015. Transthoracic two-dimensional echocardiography and color Doppler were performed for qualitative diagnosis, and the end-diastolic non-compacted layer/compacted layer (N/C) ratio measured in the parasternal ventricular short-axis sectional view was selected as the criterion for quantitative diagnosis.</p><p><b>RESULTS</b>The excessively prominent ventricular trabeculae and deep inter-trabecular recesses were all seen in 53 cases, and the blood flow in the cardiac chambers was connected to the inter-trabecular recesses. The areas involved in NCVM were mainly the apex (100%) and the middle segment of the left ventricular lateral wall (98%), followed by the middle segment of the left ventricular posterior wall (49%) and the middle segment of the left ventricular inferior wall (42%). In 53 children with NCVM, the N/C ratio was 4.3±1.9 (2.1-10.0). Cardiac insufficiency was found in 83% (44/53) of the children with NCVM, and the left ventricular ejection fraction for these children was (43±9)%.</p><p><b>CONCLUSIONS</b>Echocardiography can be used in the qualitative and quantitative diagnosis of NCVM and in the evaluation of cardiac function. The apex and the middle segment of the left ventricular lateral wall are often involved in NCVM, accompanied by decrease in the left ventricular ejection fraction.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Echocardiography , Heart Ventricles , Congenital Abnormalities , Diagnostic Imaging , Stroke Volume , Ventricular Function, LeftABSTRACT
<p><b>BACKGROUND</b>Real-time three-dimensional echocardiography (RT-3DE) has made revolutionized improvements of cardiac imaging during the past few years. However, there is no standard examination method for RT-3DE so far. This study aimed to establish the diagnostic method of RT-3DE and evaluate its application in the diagnosis of complex congenital heart diseases (CHD).</p><p><b>METHODS</b>Fifty patients with complex CHD were examined by RT-3DE with modes of Live 3DE and Full Volume. A series of novel volumetric views combined with Van Praagh sequential segmental approach were introduced to reveal the pathological morphology of the hearts, which were compared with the findings of two-dimensional echocardiography (2DE), angiography and cardiac surgery.</p><p><b>RESULTS</b>In 50 patients, 190 image acquisitions of Full Volume were performed at several acoustic windows including subcostal, apical and parasternal regions. Among them, 94.2% (179/190) of image acquisitions were successful. Most sectional volumetric views could be clearly displayed in 92.6% of the successful image acquisitions. However, sectional volumetric views could not be clearly displayed in 7.4%, which was mainly due to poor perspective conditions of examination location, improper instrument multi-parameter setting and insufficient information of whole heart captured in Full Volume acquisitions. As compared with surgical findings and angiography, RT-3DE made correction to the diagnoses in 2 cases including 1 with corrected transposition of the great arteries and the other with single atrium and mitral cleft. The diagnoses initially made by 2DE for these 2 patients were double outlet right ventricle with transposition of the great arteries and complete atrio-ventricular septal defect.</p><p><b>CONCLUSIONS</b>RT-3DE can clearly display the pathological morphology of complex CHD by a series of novel volumetric views combined with sequential segmental approach through providing more spatial informative cardiovascular structures, which provides a practical method for RT-3DE diagnosis.</p>