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Objective @#The CRISPR / Cas9 technology was applied to construct PDE4D homozygous knockout mice to provide a basis for in-depth investigation of PDE4D gene function and mechanism of action．@*Methods@#A vector was constructed for PDE4D gene exon 4，5 microinjected into fertilized eggs of C57BL /6J mice，and PDE4D －/ - mice were obtained after maternal breeding and offspring mating，and the mice genotypes were determined by PCR product sequencing and genotype identification techniques．Changes in morphology and function of the major organs of the mice were detected using an ultrasound imaging system and H＆E staining，and the expression of PDE4D protein in the mice was verified by Western blot assay. @*Results @#The PDE4D －/ － mouse genotype was stably inherited， the mice were small，and there were no obvious morphological and histological changes in the major organs in vivo. The PDE4D expression was reduced or largely absent in the major tissues of PDE4D heterozygous or pure knockout mice，and the knockout effect was better．@*Conclusion @#PDE4D －/ － mice were successfully established using CRISPR / Cas9 technology，and no significant physiological abnormalities were found，which could be used for disease pathogenesis and drug research using PDE4D as the target.
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Objective@#To retrospectively investigate the long-term swallowing function of patients with laryngeal carcinoma, who underwent partial laryngectomy, discuss the effectiveness and reliability of Kubota drinking test in the assessment of patients with dysphagia, who underwent partial laryngectomy, and analyze the influence of different ways of operation on swallowing function.@*Methods@#Clinical data were retrospectively analyzed on 83 patients with laryngeal carcinoma, who underwent partial laryngectomy between September 2012 and August 2015. Questionnaire survey, Kubota drinking test and video fluoroscopic swallowing study (VFSS) were conducted for patients during a scheduled interview. Patients were grouped by two ways: the one was whether epiglottis was retained, and the other was whether either arytenoids or both were reserved. The influence of different surgical techniques on swallowing function was analyzed according to the results of Kubota drinking test. The agreement and reliability of Kubota drinking test were statistically analyzed with respect to VFSS treated as the gold standard. SPSS23.0 software was used to analyze the data.@*Results@#Questionnaire results revealed that among 83 patients underwent partial laryngectomy 32.53% suffered from eating disorder, and 43.37% experienced painful swallowing. The incidence of dysphagia was 40.96% according to the results of Kubota drinking test. There was statistical difference between the group with epiglottis remained and that having epiglottis removed in terms of the absence of dysphagia and severity. The statistical values of normal, moderate and severe dysphagia were in the order of 18.160, 7.229, 12.344(P<0.05). Also, statistical difference existed between the groups with either and both arytenoids reserved in terms of the absence of dysphagia as well as that of intermediate severity, and their statistical values were 4.790 and 9.110(P<0.05). A certain degree of agreement and reliability was present between the results of Kubota drinking test and VFSS(Kappa=0.551, r=0.810).@*Conclusions@#It was of considerable significance to reserve epiglottis and arytenoids for the retention of swallowing function for patients post partial laryngectomy. There are certain degree of agreement and reliability between the results of Kubota drinking test and VFSS. The test, therefore, could be used as a tool for screening patients suffering from dysphagia post partial laryngectomy.
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OBJECTIVE@#To study the effect of MDR1 (P-gp) and ABCG2 on the drug resistance in Hep 2 cells.@*METHOD@#Flow cytometry was used to detect the variations of the antitumor drugs accumulation and discharging, and activity variations when MDR1 and ABCG2 inhibitors were used in Hep-2.@*RESULT@#The accumulation and discharging of mitoxantrone was significantly higher than the control group when ABCG2 inhibitor FTC was used in Hep-2 (P<0. 05). In contrast, P-gp did not appear similar case; To the mitoxantrone and cisplatin, there was no statistical correlation about activity of Hep-2 between P-gp or ABCG2 antagonist and the control; To the doxorubicin, combining FTC and P-gp, the activity of Hep-2 was higher than the control and difference was significant (P<. 05), In contrast, FTC and P-gp did not appear similar case when used alone; To the 5-FU, when PGP used, the activity of Hep-2 was higher than that in the control and difference was significant (P<0. 05), In con- trast, FTC and FTC+P-gp did not appear similar case; To the paclitaxel, when P-gp or FTC+P-gp used, the activity of Hep-2 was higher than that in the control and difference was significant(P<0. 05).@*CONCLUSION@#ABCG2 may lead to drug resistance mainly by changing the ability of cell in accumulating and discharging chemotherapy drugs. P-gp has other way. P-gp and ABCG2 play different roles in different drug resistance.