Objective To prepare humanized monoclonal antibodies(Mabs)targeting Acinetobacter baumannii(Ab)based on single memory B cell sequencing technology,construct the immune repertoire of the core protein of Ab,hemolysin-coregulated protein(Hcp),and express its Mabs with binding activity.Methods E.coli BL21 harboring the recombinant plasmid pGEX-6p-1-Hcp was constructed.Hcp protein was obtained using protein expression and affinity chromatography.Female SPF BALB/c mice(6～8 weeks old,weighing 18～20 g)were immunized intramuscularly with antigen Hcp to generate specific memory B cells.Single antigen-specific memory B cells were sorted using flow cytometry.The immune repertoire of Hcp was constructed using single-cell sequencing technology,and bioinformatics analysis was performed on the sequencing results.Mabs were obtained using antibody humanization techniques.The in vitro binding activity of the antibodies was detected by ELISA.Results The target protein Hcp with a purity>95%was obtained after expression and purification.The immune repertoire of Hcp was successfully constructed,and the results of BCR clonotype identification and analysis,CDR3 region characteristic analysis,and V-J gene pairing characteristic analysis were achieved.Antibody humanization got 7 Mabs,that is,IgG1-1,IgG1-2,IgG2-1,IgG2-2,IgG3-1,IgG4-1 and IgG4-2.ELISA results showed IgG1-1,IgG3-1,IgG4-1,and IgG4-2 had an antibody binding titer of 1∶1 280,IgG2-2 of 1∶10 240,IgG2-1 of 1∶5 120,and IgG1-2 of 1∶160.Conclusion Single-cell sequencing technology enables rapid,accurate,and efficient construction of an Hcp protein immune repertoire containing extensive antibody information.Utilizing this immune repertoire allows for the expression of Mabs with binding activity.

Objective To explore the feasibility of treating the microtia aided with the digital technology,harvesting the rib cartilage precisely,and carving fabric the ear framework in more details.Methods From July 2015 to January 2017,22 microtia patients accepted clinical therapy aided with digital technology.Preoperatively,the three-dimensional model of the costal cartilage and ear were made to design the cartilage scaffold fabrication.After the operation,the difference of the three dimensional morphology between the reconstructed ear and the normal ear was measured by digital technology.Results With average 7 months follow up,19 cases were satisfied with the outcome of the reconstructed ear including three dimensional position and subunit appearance.The outcomes were acceptable in 3 cases.There were no postoperative complications related to chest wall deformity or affected normal physical function.Conclusions Digital technology can achieve accurate harvesting and reasonable utilization of autogenous rib cartilage,provide accurate guidance for the sculpture of auricle scaffold and objective evaluation for the three-dimensional structure of the reconstructed ear.

Objective Based on the different physiological characteristics of the mastoid skin and soft tissue in congenital microtia malformation ,the different ear reconstructivemethod were respectively applied for the individuals with microtia .And the feasibility of personalized treatment to microtia patients was explored in this study .Method Considering different thickness and tightness in mastoid hairless skin and soft tissue,2129 microtia patients were received the skin expansionmethod of ear reconstruction surgery , while 1321 cases were treated with Nagata ' s technique, and 330 cases with skin and fascia expansionmethod . Result With average 10-month follow-up,1944 ( 91.31％) cases using the mastoid skin expansionmethod were satisfied with the outcome of the reconstructed ear including three dimensional position and subunit appearance.305(92.42％)cases using the skin and fascia expansionmethod were satisfied with theresult .1264 (95.69％) cases using Nagata ' s two—stagemethod received satisfactory outcomes .49 ( 3.71％) cases showed partial exposure of the cartilage framework .There were no postoperative complications , such as chest wall deformity or affected normal physical function .Conclusion Based on the different physiological characteristics of the mastoid skin and soft tissue ,it's beneficial to select individualized operationalmethod in ear reconstruction for congenital microtia among Chinese patients .

Objective Based on the different physiological characteristics of the mastoid skin and soft tissue in congenital microtia malformation ,the different ear reconstructivemethod were respectively applied for the individuals with microtia .And the feasibility of personalized treatment to microtia patients was explored in this study .Method Considering different thickness and tightness in mastoid hairless skin and soft tissue,2129 microtia patients were received the skin expansionmethod of ear reconstruction surgery , while 1321 cases were treated with Nagata ' s technique, and 330 cases with skin and fascia expansionmethod . Result With average 10-month follow-up,1944 ( 91.31％) cases using the mastoid skin expansionmethod were satisfied with the outcome of the reconstructed ear including three dimensional position and subunit appearance.305(92.42％)cases using the skin and fascia expansionmethod were satisfied with theresult .1264 (95.69％) cases using Nagata ' s two—stagemethod received satisfactory outcomes .49 ( 3.71％) cases showed partial exposure of the cartilage framework .There were no postoperative complications , such as chest wall deformity or affected normal physical function .Conclusion Based on the different physiological characteristics of the mastoid skin and soft tissue ,it's beneficial to select individualized operationalmethod in ear reconstruction for congenital microtia among Chinese patients .