Objective To investigate the structure and function of cross-links on stereocilia of guinea pig outer hair cell (OHC). Method The ultrastructures of cross-links on stereocilia of guinea pig OHC were observed by scanning electron microscopy and tannic acid procedures. Results The side-links ran horizontally between OHC stereocilia. The stereocilia of the same and different row on each hair cell were joined by horizontally-running links. The side-links were observed in the first turn as well as the fourth turn. There were more side-links on the intact stereocilia than that on the disrupted arrangement. The side-links in the first turn and the fourth turn were more abundant than that in the second turn and the third turn. The side-links between stereocilia were spare if the stereocilia were separated. The number of side-links on stereocilia was proportional to the number of bulb-like structures on stereocilia. Conclusion The side-link is a kind of morphological structure on hair cells stereocilia of cochlea. The results suggest that cross-links play an important role in maintaining the structure and function of the hair cell stereocilia.

This paper presented the hereditary pattern of otosclerosis in families. Among 1500 cases there were 13 patients, belonging to 6 families, and 20 ears were surgically treated. There was often a marked similarity in clinical features of affected siblings within the same family.The clinical characteristics are as following: (1) The age at inset of hearing loss seems similar. There is a difference of 1 or 2 years in two families znc 3 to 10 years in the other four. (2) The side of first hearing loss is the same in affected individuals of 6 families. (3) The propositi are females in 5 families (83.3%). (4) Auditory curve of hearing loss is similar in 4 familiers, and different in the other two. (5) The footplate lesions are similar in macroscopic features, thin footplate in two fimalies and thick footplate in the other four.

Objective:To construct and purify four respiratory syncytial virus (RSV) PreF proteins through gene sequence design and optimization and evaluate their immunogenicity.Methods:Coronin-1A and T4 trimer protein gene sequences were optimized with Human and CHO codons, and then added to RSV F protein sequence. The above plasmids were transfected into Expi293F cells for protein expression. After purification by nickel column, four trimer proteins were prepared. SDS-PAGE and Western blot were performed for protein identification. BALB/c mice were immunized at week 0 and week 3, and blood samples were collected to measure the activities of binding and neutralizing antibodies in serum.Results:SDS-PAGE and Western blot showed that the four proteins had stable trimer structure. Antigen-antibody affinity test showed that the four trimer proteins had strong affinity with RSV-specific monoclonal antibodies 8897, D25, Motavizumab, AM14 and Palivizumab. The titers of antibodies induced by the two T4 trimers were higher after the initial immunization, while there was a substantial increase in the titers of antibodies induced by Human codon-optimized trimer protein after the second immunization.Conclusions:PreF trimer protein can be prepared by adding any of the two different heterotrimer motifs, and induce effective binding and neutralizing antibodies in mice.

Objective:To compare the immunogenicity of the prefusion (PreF) and postfusion (PostF) conformations of the respiratory syncytial virus (RSV) F protein.Methods:The expression of PreF and PostF recombinant proteins was analyzed by SDS-PAGE and Western blot. The binding affinity between F protein and its specific antibodies was detected by Octet. The binding antibodies and neutralizing antibodies in immune serum were detected after immunizing mice with PreF or PostF recombinant protein.Results:PreF protein was stable in the form of a trimer after modification with higher binding affinity with monoclonal antibodies such as D25, 8897, AM14, Palivizumab and Motavizumab. PostF protein lacked the antigenic site ? and showed a monomer conformation. Besides, it was unable to bind to D25, 8897 and AM14 antibodies. Animal experiments showed that AS01 adjuvant was better than aluminum adjuvant in inducing binding antibodies and neutralizing antibodies against RSV Long strains. The binding antibodies induced by PreF and PostF recombinant proteins had similar binding ability to PreF protein, while the binding antibodies induced by PostF recombinant protein showed stronger binding ability to PostF than to PreF.Conclusions:PreF has more epitopes and the trimer form of PreF recombinant protein after modification is more stable and can induce stronger neutralizing antibodies. Moreover, the immunopotentiating effect of AS01 adjuvant is better than that of aluminum adjuvant. Therefore, stabilization-based trimer structure modification of PreF and the development of adjuvants are crucial for the development of RSV vaccines.