The aim of this study was to assess changes of Hsp70 and HSF-1 protein and mRNA expression in stress-sensitive organs of pigs during transportation for various periods of time. Twenty pigs were randomly divided into four groups (0 h, 1 h, 2 h, and 4 h of transportation). A significant increased activity of AST and CK was observed after 1 h and 2 h of transportation. Histopathological changes in the heart, liver, and stomach indicated that these organs sustained different degrees of injury. Hsp70 protein expression in the heart and liver of transported pigs did not change significantly while it increased significantly (p < 0.05) in the stomach. Hsp70 mRNA levels decreased significantly (p < 0.05) in the heart after 4 h of transportation. However, mRNA expression increased significantly in the liver after 1 (p < 0.05) and 4 h (p < 0.01) of transportation, and increased significantly in the stomach of the transported pigs after 1, 4 (p < 0.01), and 2 h (p < 0.05). HSF-1 levels were reduced at 1 and 4 h (p < 0.05) only in the hearts of transported pigs. These results indicate that Hsp70 mediates distinct stress-related functions in different tissues during transportation.
Animals ; Creatine Kinase/blood ; DNA-Binding Proteins/*metabolism ; Enzyme-Linked Immunosorbent Assay/veterinary ; HSP70 Heat-Shock Proteins/*metabolism ; Liver/*metabolism ; Myocardium/*metabolism ; RNA, Messenger/metabolism ; Random Allocation ; Real-Time Polymerase Chain Reaction/veterinary ; Stomach/*metabolism ; Stress, Physiological ; Swine/blood/*metabolism ; Time Factors ; Transaminases/blood ; Transcription Factors/*metabolism ; *Transportation

Objective:To investigate the effect of repeated freezing and thawing on the clinical outcome of embryo transfer.Methods:A total of 48 cycles of twice frozen-thawed embryo (blastocyst) transfer in the reproductive medicine department, Shenzhen Luohu People′s Hospital from 2015 to 2020 were collected as the observation group, and 98 cycles of one frozen-thawed blastocyst transfer in the same period were randomly selected as the control group. The patient′s age, endometrial thickness, average number of transferred embryos, average number of high-quality embryos transferred, embryo recovery rate, implantation rate, clinical pregnancy rate, abortion rate, ectopic pregnancy rate, and live birth rate were compared.Results:There was no significant difference in age, endometrial thickness, number of embryos transferred and high-quality embryos transferred between the two groups (all P>0.05). There was no significant difference in embryo recovery rate, ectopic pregnancy rate, abortion rate and live birth rate between the two groups (all P>0.05). The embryo implantation rate and clinical pregnancy rate in the observation group were significantly lower than those in the control group (32.47% vs 55.70%, 39.58% vs 66.33%, all P<0.05). Conclusions:The clinical pregnancy rate was significantly lower than that of the control group after repeated vitrification of frozen-thawed embryo transfer, which may affect the subsequent developmental potential of embryos.

Binding sites of five monoclonal antibodies were obtained by reinforceable method of overlapping recombinant prion protein and synthetic peptide. Overlapping peptides of PrP core were expressed in Escherichia coli by insertion of serial PCR amplicons of ovine PrP gene fragments into pET32a. The expressed fusion peptides were then tested for the binding activity to PrP monoclonal antibodies in Western blotting. The binding sites of 5 monoclonal antibodies of ovine PrP were located respectively as follows: 2H3 in 199 aa-213 aa, 4C6, 5F11 and 7F11 in 139 aa-168 aa and 7F1 in 214 aa-227 aa. There oligo peptides were synthesized and used in ELISA test for more accurate localization of the binding sites. The binding sites of 4C6, 5F11 and 7F11 were further confirmed to be in 149 aa-158 aa. This conclusion may contribute to the research for pathogenesis and diagnostic method of scrapie and bovine transmissible spongiform encephalopathy.
Animals ; Antibodies, Monoclonal ; immunology ; metabolism ; Binding Sites, Antibody ; immunology ; Epitopes ; immunology ; Escherichia coli ; genetics ; metabolism ; Prion Diseases ; diagnosis ; Prions ; genetics ; immunology ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Scrapie ; diagnosis ; Sheep