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Objective:To observe the inhibitory effects of Huangqi Jiedu Decoction on lung metastasis of breast cancer in nude mice; To explore the mechanism of intervening epithelial mesenchymal transformation (EMT) induced by Wnt/β-catenin signaling pathway.Methods:Totally 30 nude mice were divided into model group, adriamycin group and Huangqi Jiedu Decoction low-, medium-, and high-dosage groups according to random number table method. Each group was injected subcutaneously with mouse breast cancer 4T1 cells to construct tumor - bearing nude mice model. Huangqi Jiedu Decoction low-, medium- and high-dosage groups were intragastrically administrated with Huangqi Jiedu Decoction 17.82, 35.64 and 71.28 g/kg; adriamycin group was injected intraperitoneally adriamycin 0.05 g/kg; model group was intragastrically administrated with normal saline of the same volume for 21 d. Tumor volume was measured at 9, 15, and 21 days after modeling. After the end of administration, the tumor tissue was separated, the tumor weight was measured, and the tumor inhibition rate was calculated. The lung tissue was Isolated,, the number of lung metastatic nodules and the inhibition rate of lung metastasis was counted. HE staining was used to observe the tissue morphology and evaluate the effectiveness of the model. The protein expressions of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by Western Blot. The mRNA levels of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the model group, the tumor volume and mass of Huangqi Jiedu Decoction low-, medium- and high-dosage groups decreased ( P<0.01); the number of pulmonary metastasis nodules in Huangqi Jiedu Decoction high-dosage group significantly decreased ( P<0.01); the mRNA and protein expressions of β-catenin and Vimentinm decreased in the Huangqi Jiedu Decoction low-, medium- and high-dosage groups ( P<0.01), and the protein and mRNA expressions of E-Cadherin increased in the Huangqi Jiedu Decoction high-dosage group ( P<0.01). Conclusion:Huangqi Jiedu Decoction can effectively inhibit the growth and lung metastasis of breast cancer transplanted tumor, and the mechanism may be to down-regulate the expression of key molecules in the Wnt/β-catanin signaling pathway, thereby inhibiting the EMT process, so as to inhibit the lung metastasis of breast cancer.
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Background:Fusobacterium nucleatum (Fn)is a common oral pathogen. Studies have shown that Fn is closely related to the occurrence and development of colorectal cancer,especially the inflammation-related colorectal cancer. Aims:To investigate the mechanism of Fn in forming an inflammatory microenvironment in colon cancer cells. Methods:An inflammation model of Caco-2 cells infected by Fn was constructed,and miRNA sequencing was performed. miR-181b mimics or inhibitor was transfected into Fn infected Caco-2 cells. mRNA and protein expressions of TNF-α were determined by qRT-PCR and Western blotting,respectively,and concentration of TNF-α in supernatant was measured by ELISA, number of lymphocyte penetrating the membrane was measured by Transwell chamber. Results:Compared with control group,mRNA and protein expressions of TNF-α were significantly increased (P < 0. 05),concentration of TNF-α in supernatant was significantly increased (P < 0. 05),and number of lymphocyte penetrating the membrane was significantly increased in Fn group (P < 0. 05). miRNA sequencing and qRT-PCR results showed that expression of miR-181b was significantly decreased in Fn group than in control group (P < 0. 05). Compared with control group,mRNA and protein expressions of TNF-α were significantly decreased in miR-181b mimics + Fn group (P < 0. 05),however,mRNA and protein expressions of TNF-α were significantly increased in miR-181b inhibitor group (P < 0. 05). Bioinformatics tools and Luciferase assay confirmed that TNF-α might be the target gene of miR-181b in Caco-2 cells. Conclusions:Fn can up-regulate the expression of TNF-α by inhibiting miR-181b in Caco-2 cells and recruiting lymphocytes to form an inflammatory microenvironment.
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Background:Fusobacterium nucleatum (Fn)is a common oral pathogen. Studies have shown that Fn is closely related to the occurrence and development of colorectal cancer,especially the inflammation-related colorectal cancer. Aims:To investigate the mechanism of Fn in forming an inflammatory microenvironment in colon cancer cells. Methods:An inflammation model of Caco-2 cells infected by Fn was constructed,and miRNA sequencing was performed. miR-181b mimics or inhibitor was transfected into Fn infected Caco-2 cells. mRNA and protein expressions of TNF-α were determined by qRT-PCR and Western blotting,respectively,and concentration of TNF-α in supernatant was measured by ELISA, number of lymphocyte penetrating the membrane was measured by Transwell chamber. Results:Compared with control group,mRNA and protein expressions of TNF-α were significantly increased (P < 0. 05),concentration of TNF-α in supernatant was significantly increased (P < 0. 05),and number of lymphocyte penetrating the membrane was significantly increased in Fn group (P < 0. 05). miRNA sequencing and qRT-PCR results showed that expression of miR-181b was significantly decreased in Fn group than in control group (P < 0. 05). Compared with control group,mRNA and protein expressions of TNF-α were significantly decreased in miR-181b mimics + Fn group (P < 0. 05),however,mRNA and protein expressions of TNF-α were significantly increased in miR-181b inhibitor group (P < 0. 05). Bioinformatics tools and Luciferase assay confirmed that TNF-α might be the target gene of miR-181b in Caco-2 cells. Conclusions:Fn can up-regulate the expression of TNF-α by inhibiting miR-181b in Caco-2 cells and recruiting lymphocytes to form an inflammatory microenvironment.
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Objective To evaluate the prospective electrocardiograph (ECG)-gated 128-slice spiral CT angiography in the diagnosis of complex congenital heart disease and airway abnormality in children.Methods Eighty children with congenital heart disease received prospective (n =40) or retrospective ECG-gated 128-slice spiral CT angiography (n =40).All of the patients underwent ultrasound cardiogram (UCG) within one week before CTA exmination.The image quality and the effective radiation dose were statistically analyzed using independent samples t test and x2 test.The surgical results were taken as the diagnostic standards,and the preoperative prospective and retrospective CT angiography data were compared with UCG results using x2 test.Results The effective dose of prospective ECG-gated CT angiography[(1.00 ± 0.26)mSv] was reduced by 86％ as compared with the retrospective ECG-gated CT angiography[(7.37 ± 0.52) mSv] (t =69.296,P ＜ 0.01).Image quality was graded 0 in none patients,graded 1 in 2 patients who underwent prospective ECG-gated CT angiography,graded 2 in 8 and 1 patient who received prospective and retrospective ECG-gated CT angiography respectively,and graded 3 in 30 and 39 patients who underwent prospective and retrospective ECG-gated CT angiography,respectively.There was no significant difference in image quality between prospective and retrospective ECG-gated CT angiography (x2 =0.044,P ＞ 0.05).The definitive diagnosis of extracardiac vascular anomalies was made in 98.3％(59/60) of patients by using prospective and retrospective ECG-gated CT angiography,which were significantly superior to UCG results[(76.7％ (46/60),76.7％ (45/59),respectively] (x2 =13.144,12.644,P ＜ 0.01),and there was no significant difference in the definitive diagnosis between prospective and retrospective ECG-gated CT angiography (x2 =0.001,P ＞ 0.05).The definitive diagnosis of the cardiac/large-vessel malformations was made in 100％ of the patients by both prospective and retrospective ECG-gated CT angiography,and there was no significant difference between 128-slice spiral CT angiography results and UCG results [97.2％ (35/36),97.3％ (36/37),respectively] (x2 =1.014,0.001,P ＞0.05).In the cardiac malformations,the definitive diagnosis by prospective and retrospective ECG-gated CT angiography was 88.7 ％ (47/53) and 92.5 ％ (49/53),and there was no significant difference between 128-slice spiral CT angiography results and UCG results [98.1％ (52/53),98.1％ (52/53),respectively](x2 =3.824,1.889,P ＞ 0.05),and no significant difference in the definitive diagnosis was observed between prospective and retrospective ECG-gated CT angiography (x2 =0.442,P ＞ 0.05).In addition,the 128-slice spiral CT angiography detected airway abnormality in 9 cases who underwent prospective or retrospective ECG-gated CT angiography.Conclusion The prospective ECG-gated 128-slice spiral CT angiography exhibits great application value in the preoperative diagnosis of complex congenital heart disease,which helps to develop surgical program and evaluate surgical risks.