This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.

This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.

Objective To analyze the effect of long-term wearing of hard corneal contact lens and its effect on the stability of tear film.Methods From April 2016 to April 2017,150 patients of myopia in Yongkang Hospital were selected in this study and randomly divided into study group and control group,with 75 cases in each group.The patients in the study group wore hard corneal contact lens for a long time before operation,the patients in the control group had no hard corneal contact lens before operation.The basal tear secretion amount,tear film break-up time(TBUT) and the incidence of postoperative complications were compared between the two groups.Results One year after operation,the basal tear secretion amount of the study group [(11.1 ± 1.5) mL] was significantly lower than that of control group[(18.5 ± 3.2) mL] (t =12.235,P ＜ 0.05).The TBUT of the study group [(15.8 ± 1.5) min] was significantly shorter than that of the control group [(25.1 ± 4.8) min] (t =11.935,P ＜ 0.05).The incidence rates of dry eye,corneal staining in the study group were 22.7％,18.7％,respectively,which were significantly lower thanthose in the control group (33.3％,34.7％),there were statistically significant differences between the two groups (x2 =5.931,6.425,all P ＜ 0.05).Conclusion Long-term wearing hard contact lens seriously affects the tear secretion and TBUT,and reduces the stability of tear film.Although the incidence of postoperative complications islow,tear film damage can not be avoided.In order to protect the stability of the tear film,the patients with myopia should try to avoid the wear of the hard corneal contact lens and choose to wear frame glasses.

Diabetes mellitus( DM)affects almost the whole digestive tract. Approximately 75% of the diabetic patients suffer from gastrointestinal motility disorders. The precise mechanism of diabetic gastrointestinal motility disorders is not fully clear. Its pathogenesis is considered to be multifactorial and related with autonomic neuropathy,smooth muscle myopathy and lesions of interstitial cells of Cajal. Advanced glycation end products( AGEs ) are derived from the nonenzymatic reaction of glucose with proteins,lipids or nucleic acids in vivo. Hyperglycemia in DM is a favourite for formation of AGEs, and excessive accumulation of AGEs contributes to the complications of DM such as diabetic nephropathy and diabetic vascular lesions. However,the correlation of AGEs with diabetic gastrointestinal motility disorders is seldom reported. In this review article,the roles and possible mechanisms of AGEs in diabetic gastrointestinal motility were summarized.