Experimental study of chondrogenesis in vitro by co-culture of bone marrow stromal cells and chondrocytes / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of chondrogenesis in vitro with bone marrow stromal cells (BMSCs) induced by the co-cultured chondrocytes.</p><p><b>METHODS</b>The BMSCs and chondrocytes were separated from pig and cultured. The supernatant of chondrocytes was used as the inducing solution for BMSCs from the 2nd generation. 7 days later, samples were taken and underwent immunohistochemistry and RT-PCR for detection of the expression of specific type II cartilage collagen, type II collagen and aggrecan mRNA. The cultured BMSCs and chondrocytes were mixed at a ratio of 8:2 (BMSC: cartilage cell) and were inoculated into a polyglycolic acid/polylactic acid (PGA/PLA) scaffold at the final concentration of 5.0 x 10(7)/ml. The cartilage cells and BMSCs were also inoculated separately at the same concentration as the positive and negative control. Pure cartilage cells at 20% of the above mentioned concentration (1.0 x 10(7)/ml) were used as the low concentration cartilage cell control group. Samples were collected 8 weeks later. General observations, wet weight, glycosaminoglycans (GAGs) determination and histological and immunohistochemistry examinations were performed.</p><p><b>RESULTS</b>The expression of type II collagen, type II collagen and aggrecan mRNA were positive in induced BMSCs. In the co-cultured group and the positive control group, pure mature cartilage was formed after 8 weeks of culture in vitro, and the size and shape of the scaffold were maintained. The newly formed cartilage in the two groups were almost the same in appearance and histological properties. The immunohistochemistry results indicated that the cartilage cells of the two groups all expressed ample cartilage-specific type II collagen. The average wet weight and GAG content in the co-cultured group reached more than 70% of those in positive control group. Only an extremely small amount of immature cartilage tissues was formed in local regions in pure BMSC group, and the scaffold was obviously shrunk and deformed. Although the wet weight of newly generated cartilage tissue in the low concentration cartilage cell group reached 30% of that in positive control group, the scaffold was obviously shrunken and deformed. Only regional and discontinuous cartilage tissues were formed, and the amount of newly formed cartilage was obviously less than that in the co-culture group and the positive control group.</p><p><b>CONCLUSIONS</b>Chondrocytes can provide a micro-environment for the formation of cartilage, and also effectively induce BMSC to differentiate into chondrocytes and form tissue-engineered cartilage in vitro.</p>

The mutation study of the FOXL2 gene in a big Chinese family with blepharophimosis-ptosis-epicanthus inversus syndrome / 中华整形外科杂志

<p><b>OBJECTIVE</b>We have studied 4 generations 12 patients in a family which has blepharophimosis-ptosis-epicanthus-inversus syndrome (BPES) for the gene, FOXL2, the group also have 12 normal members in this family and other 80 normal individuals for contrast.</p><p><b>METHODS</b>The FOXL2 gene was amplified by polymerase chain reaction and then analyzed by direct genomic sequencing.</p><p><b>RESULTS</b>A 892C > T at nucleotides in FOXL2 was found in the twelve affected patients. No mutations was found in any of the health members in the family.</p><p><b>CONCLUSIONS</b>FOXL2 may be a important pathogenesis for the disease in this Chinese family.</p>

An observation of the effects of the truncated septin2 on mouse epidermal cell and fibroblast / 中华整形外科杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression vector of the truncated septin2 and investigate the influence on the cultured mouse epidermal cell and fibroblast in vitro exerted by the transgenic expression product.</p><p><b>METHODS</b>The short splicing fragment was obtained by amplifying the reverse transcription product of the fetal mouse skin mRNA with PCR. Then its recombinant expression vector pcDNA3.1 (-)/septin2s was constructed and used to transfect the mouse epidermal cell and fibroblast cultured in vitro. The expression of the foreign gene was detected with RT-PCR and the changes of cell proliferation were observed and analysed.</p><p><b>RESULTS</b>RT-PCR results indicated that pcDNA3.1/septin2 was expressed in the cultured mouse epidermal cells and fibroblasts in vitro. We found that the epidermal cells accelerated their reproduction, but the fibroblasts had no obvious changes.</p><p><b>CONCLUSION</b>We successfully constructed eukaryotic expressive vectors of pcDNA3.1/ septin2s and transfected it into mouse epidermal cells and fibroblasts in vitro. The results settle a basis for showing effect of septin2s on fetal mouse skin.</p>

Template design of tissue flaps for covering auricular cage in one-stage total auricular reconstruction / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the template design of tissue flaps for covering auricular cage in order to acquire accurate and reliable design method.</p><p><b>METHODS</b>By the theory of engineering drawing and three-dimensional measuring on CT image, three dimensional configuration of 40 auricular surfaces were expanded approximately, and the character of them was analysed for the template design.</p><p><b>RESULTS</b>It is similar of the expanded graphs of auricular surface three dimensional configuration in healthy persons, and simplified template of tissue flaps is drawn based on the key points of the above graph.</p><p><b>CONCLUSIONS</b>CT three-dimensional measurement of auricular surface configuration can be used to design the template of tissue flaps for covering auricular cage, and can provide accurate and reliable template of tissue flaps for clinics.</p>

The effect of pcDNA 3.1/RPs15 on skin fibroblasts in vitro / 中华整形外科杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression vector of ribosomal protein sl5(RPs15) gene and study its effect on mouse skin fibroblasts in vitro.</p><p><b>METHODS</b>The RPs15 cDNA encoding region of fetal mouse skin was amplified by reverse transcription-polymerase chain reaction (RT-PCR) method and cloned into eukaryotic expression vector pcDNA3.1(-). The recombinant plasmid was transfected into adult mouse skin fibroblasts by FuGENE6 transfection reagents. Then the expression of RPs15 gene, was detected and its biological effect on fibroblasts was measured.</p><p><b>RESULTS</b>The DNA sequencing result of pcDNA3.1/RPs15 was identical with the reported. The RPs15 gene was expressed in transfected fibroblasts. The growth density of fibroblasts decreased with the conformation changing accordingly.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector pcDNA3.1/RPs15 is successively constructed and can be expressed in mouse skin fibroblasts. The results set up a basis for further study of the effect of RPs15 gene on skin fibroblasts.</p>

A novel mutation in the FOXL2 gene in a Chinese family with blepharophimosis, ptosis, and epicanthus inversus syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen mutations in the forkhead transcriptional factor 2 gene (FOXL2) in six Chinese families with blepharophimosis, ptosis, and epicanthus inversus syndrome(BPES).</p><p><b>METHODS</b>PCR amplification and direct sequencing of the FOXL2 coding region in genomic DNA were performed in affected patients and 80 healthy controls. BLAST analysis of the sequence was made on Internet.</p><p><b>RESULTS</b>A novel 951-953(delC) was found in the two affected patients of a Chinese family with BPES. No mutations were found in the healthy controls. The 951-953(delC) may cause a frameshift mutation after codon 238 that exists downstream of the forkhead domain, resulting in the production of truncated proteins.</p><p><b>CONCLUSION</b>These findings indicated that the 951-953(delC) deletion mutation in the two patients resulted in truncated proteins and hence led to their BPES. To the authors' knowledge, the 951-953(delC) in FOXL2 has not been previously reported.</p>