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Li X.-L.,Shenzhen Nan Shan Hospital | Yi W.-H.,Shenzhen Nan Shan Hospital | Jin A.-M.,Southern Medical University | Min S.-X.,Southern Medical University
Chinese Journal of Tissue Engineering Research | Year: 2015

BACKGROUND: In literatures, the recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded microcapsules can be fabricated by double emulsion solvent evaporation method with different organic solvents, such as methylene chloride, ethyl acetate or their mixture. But so far it is not determined yet which one is better. OBJECTIVE: To optimize the preparation method of microcapsules encapsulating rhBMP-2 and to compare the effects of different organic solvents on the microcapsules. METHODS: Polylactic acid-polyethylene glycol-polylactic acid copolymer as microcapsules was used to prepare rhBMP-2 loaded microcapsules with double emulsion solvent evaporation method. Four kinds af arganic solvents, methylene chloride (group A), mixture of methylene chloride and ethyl acetate (group B), ethyl acetate (group C) and acetyl acetone (group D) were chosen as ail phases tt compare their effects an microcapsule’s morphology, diameter, and encapsulation efficiency. Passage 3 bone marrow mesenchymal stem cells from rats were co-cultured with prepared microcapsules for 14 days, and then alkaline phosphatase activity was detected. RESULTS AND CONCLUSION: Compared with the other organic solvents, dichloromethane could cause microcapsules with the smaller and more uniform shape (4-10 microns) and the highest encapsulation efficiency; the microcapsules prepared by mixture of methylene chloride and ethyl acetate had relatively wide size distribution and moderate ancapsulation efficiincy; the microcapsuls prepared by acetylacetone were difficult to form and keep the bioactivity of rhBMP-2. After cultured with rat bone marrow mesenchymal stem cells for 14 days, the alkaline phosphatase activity in groups A, IB and C was significantly higher than that in group D and there was no significant difference between group A and group IB; the alkaline phosphatse activity in groups A and B was significantly higher than that in group C (P < 0. 05). The results show the rhBMP-2-loaded microcapsules prepared by methylene chloride as organic solvent have good shape, high ancapsulation efficiency, and good biological activity. © 2015, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.

Yu Z.-Q.,Huazhong University of Science and Technology | Hu F.-L.,Shenzhen Nan Shan Hospital | Cheng Q.,Shenzhen Nan Shan Hospital | Hao J.-H.,Shenzhen Nan Shan Hospital | And 6 more authors.
Journal of Huazhong University of Science and Technology - Medical Science | Year: 2015

This study looked into a family involving a rare mother-child ABO blood type inconsistency and explored its genetic and molecular basis. In the family, the mother had type AB blood and the father was blood type B and they gave birth to a baby of blood type O. Their blood types were phenotypically identified by using different techniques, including micro-column gel test, immune inhibition test, absorption and elution tests. The sequences of all 7 exons of ABO allele from the core family members were determined by using PCR and clone-based sequencing. The loci of mutated gene were compared against normal human genes. The result showed that the mother’s erythrocytes were agglutinable with monoclonal anti-A antibody (2+) and had agglutination reaction with anti-B antibody (4+). The mother’s serum registered agglutination action with standard blood type A cells. The findings showed an ABO inconsistency. When domestic antibodies were used, the mother’s erythrocytes yielded agglutination reaction with humanized anti-B serum (4+) and anti-B monoclonal antibody but were non-agglutinable with humanized anti-A serum and anti-A monoclonal antibody. Upon absorption and elution, the titer of anit-A antibody was 128 both before and after the absorption test, with no significant difference found between pre- and post-absorption values. Our results confirmed that the mother’s allelic gene was type B and contained type A. The father’s blood type was type B, and son’s blood type was type O. Clone-based sequencing revealed that the mother carried a heterozygous gene of B101.01 (ntA640→G)/O01, which contained an M214→V mutation that could express a weak expression of antigen A, resulting in blood type AB. However, their son did not have the M214→V mutation, which yielded a false ABO-inconsistency between him and his mother. We were led to conclude that type B gene with a M214→V mutation can encode both antigen B and weak antigen B that can lead to false ABO-inconsistencies. © 2015, Huazhong University of Science and Technology and Springer-Verlag Berlin Heidelberg.

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