The aim is to explore and build the treatment-care and four linkage combination integrated care model,so as to provide methods and ideas for the medical service practice.By exploring the mechanism and hierarchical management mode of the treatment-care and four linkage combination integrated care model,we have set up a whole process management closed loop in four combined linkage,including hospital,medical care,community and family,raised to solve the medical combination of acute medical treatment and the problems existing in the continuing care.After more than a year of preparation and practice,we have build the social service centered medical combined with four linkage information platform,and have realized the online and offline medical combination of care services in hospital,medical care,community and family.Based on Intemet technology,the building of the treatment-care and four linkage combination integrated care model is the efficient method to solve the difficulties and problems of the health care and the continuation demands among elderly under the current conditions,and the feasible way to realize theslight illness in the community,disease the hospital,care in institutions,pension in the family.

To construct TeI3c/4c-based and temperature-inducible gene inactivation system (Thermotargetron) and to apply it to gene inactivation of mesophilic bacteria. The subunit of flagellum (fliC) and C4 dicarboxylate orotate:H⁺ symporter (dctA) genes were chosen as targets in the genome of Escherichia coli HMS174 (DE3) strain. According to recognition roles of TeI3c/4c intron, the fliC489a, fliC828s, fliC1038s and dctA2a sites were chosen as target sites. Gene-targeting plasmids were constructed based on pHK-TT1A by using overlap PCR method and transformed into HMS174 cells. An aliquot mid-log phase cultures of the transformants were shocked at 48 °C and plated on LB plate (containing chloramphenicol). Afterwards, gene mutants were screened by using colony PCR and DNA sequencing. After the mutants were obtained, the phenotypes of ΔfliC and ΔdctA gene mutants were characterized by using agar puncture and carbon metabolism experiments. Colony PCR and sequencing results show that TeI3c/4c intron was inserted in the designed sites of fliC and dctA genes. The gene-targeting efficiency of Thermotargetron system was 100%. Phenotype verification experiments of the mutants demonstrated that the cell motility of all ΔfliC mutants was damaged and the malate assimilation ability of ΔdctA mutant was deprived comparing to wild-type HMS174 strain. In our study, a temperature-inducible and high-efficiency gene inactivation system was established for mesophilic bacteria. This system could achieve high efficiency and precise gene inactivation by modulation of the incubation duration of the transformants at 48 °C.