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Objective The aims of this study were to construct short hairpin RNA(shRNA)lentiviral vector in breast cancer T47D cells,to carry out RNA interference on lysine acetyltransferase 6B(KAT6B/MORF)gene,to down-regulate its expression and to explore its function.Methods Two pairs of single-stranded short hairpin RNA(shRNA5 and shRNA8)and the corresponding control sequences(Scramble5 and Scramble8)were synthesized based on the CDS of KAT6B gene.Polymerase chain reaction(PCR) was used to amplify double-stranded and ligated with the entry vector(pENTR/pSM2(CMV)GFP),which were subjected to a doub-le digestion(EcoRl and Xhol)linearization and homologous recombination with the entry vector(pENTR/pSM2(CMV)GFP)to obtain an entry clone containing the desired fragment.The target fragment was recombined onto the target vector(pLenti x1 puro DEST)via the LR cloning reaction of the Gateway system.The lentiviral packaging plasmids were co-transfected into HEK-293T cells with two pairs of target plasmids. The supernatant of HEK -293T cells was collected and transformed into T47D cells. The expression of KAT6B protein was detected in T47D cells by Western blot.Results The single colony obtained from the transformation was identi-fied by sequencing,which was consistent with the target sequence,indicating that the lentiviral vector had been successfully construc-ted.The expression of KAT6B protein was significantly lower in the shRNA KAT6B group than that in the control group,which indica-ted that the constructed gene silencing vector could play a role in the KAT6B gene in T47D cells.Conclusion The shRNA lentiviral gene silencing vectors of KAT6B were constructed and identified in T47D cells,which indicated that the foundation for further study
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Objective To study the feasibility of noninvasive detection of unstable plaques with ~(18)F-Fluorodeoxyglu-cose (~(18)F-FDG) PET/CT imaging. Methods Atherosclerotic plaques were induced in male New Zealand white rabbits. Animals were injected with FDG labeled with ~(18)F, then examined with PET/CT. Aorta was explanted for photography with digital camera, and ~(18)F-FDG uptake analysis. Thirty unstable plaques and 30 stable plaques were choosed so as to compare the quantitativly ~(18)F-FDG uptake. The number of macrophages and smooth muscle cells was detected by immunohistochemical technique. Results Experimental group showed inconsistent uptake of ~(18)F-FDG in the abdominal aorta. The results were confirmed in the ex vivo digital photo of the explanted aorta. The target to non target ratio (T/NT) and macrophages of unstable plaques were higher than stable plaques (P<0.01) , but smooth muscle cells obviously reduced (P <0. 01). Correlation analysis showed that there was a positive correlation between T/NT and macrophage content (r=0. 815,P<0. 01), and a negative correlation between T/NT and SMC content(r=-0. 684,P <0. 01). Conclusion ~(18)F-FDG PET/CT can constitute an attractive imaging method for the noninvasive detection of experimental unstable plaques.
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Objective To study the feasibility of noninvasive detection of unstable plaques with 18F-Fluorodeoxyglucose (18F-FDG) PET/CT imaging. Methods Atherosclerotic plaques were induced in male New Zealand white rabbits. Animals were injected with FDG labeled with 18F,then examined with PET/CT. Aorta was explanted for photography with digital camera,and 18F-FDG uptake analysis. Thirty unstable plaques and 30 stable plaques were choosed so as to compare the quantitativly 18F-FDG uptake.The number of macrophages and smooth muscle cells was detected by immunohistochemical technique. Results Experimental group showed inconsistent uptake of 18F-FDG in the abdominal aorta. The results were confirmed in the ex vivo digital photo of the explanted aorta. The target to non target ratio(T/NT) and macrophages of unstable plaques were higher than stable plaques(P
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Objective To research the mechanism of treating cyclomastopathy with Runingshuang by measuring the blood rheology and mammary microcirculation. Methods 70 female Wistar rats of SPF were randomly divided into seven groups:the blank control group, the pathologic model group, the comparative group of tamoxifen ointment, the comparative group of Sanjierupi Glue, three remedial groups with large, middle and small dosage of Runingshuang. Pathologic model of mammary hyperplasia was made by injecting estrogen and progesterone. Except the former two groups, each group was treated with different drugs. The blood rheology and mammary microcirculation were measured after 30 days. Results Compared with the blank control group, blood viscosity, plasma viscosity, packed cell volume and the fibrinogen of the pathologic model group increased evidently (P
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OBJECTIVE:To study the anti-inflammation,analgesia and microcirculation improvement actions of Runingshuang.METHODS:The animals were randomly divided into control group,Sanjierupi glue group and lose dose and high dose Runingshuang groups.The curative effects of the externally applied Runingshuang on dimethylbenzene-induced earlap turgescence and the agar-induced chronic granulomatous inflammation were monitored.And the analgesic effect of the intragastrically administered or externally applied Runingshuang and its antagonistic action in microcirculation disturbance caused by pituitrin were determined by glacial acetic acid writhing method and hot plate method.RESULTS:The inhibitory rate of high and low dosage of Runingshuang on the earlap turgescence caused by dimethylbenzene were 46.2%and 43.1%,respectively,which were significantly different as compared with the control group(P
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Objective To study the relation between mammary hyperplasia and expression of VEGF,bFGF. Methods After the breast mass undergoing core needle biopsy, the pathological type and expression of VEGF and bFGF were observed in 140 cases of breast mass. Results Of the 140 cases, general ~hyperplasia was found in 92 cases( 65.7%), atypical hyperplasia in 48 cases(34.3%) . The expression of VEGF and bFGF were increased with the increase of pathological degree of breast hyperplasia (P