Genotype-Phenotype Relationship and Follow-up Analysis of a Chinese Cohort With Osteogenesis Imperfecta.

OBJECTIVE: To evaluate the genotype-phenotype relationship and the effect of treatment on the clinical course of osteogenesis imperfecta (OI). METHODS: We established a Chinese hospitalized cohort with OI and followed them up for an average of 6 years. All patients were confirmed as having OI using whole-exome sequencing. We analyzed the genotype-phenotype relationship based on different types, pathogenic mechanisms, and gene inheritance patterns of OI. Additionally, we assessed whether there was a difference in treatment efficacy based on genotype. RESULTS: One hundred sixteen mutations in 6 pathogenic genes (COL1A1, COL1A2, IFITM5, SERPINF1, FKBP10, and WNT1) were identified in 116 patients with type I, III, IV, V, VI, XI, or XV OI. Compared with patients with COL1A1 mutations, patients with COL1A2 mutations were younger at the time of the first fracture, whereas other phenotypes were similar. When 3 groups (helical, haploinsufficiency, and non-collagen I gene mutations) were compared, patients with helical mutations were the shortest and most prone to dentinogenesis imperfecta. Patients with haploinsufficiency mutations were the oldest at the time of the first fracture. Moreover, patients with non-collagen I gene mutations were least susceptible to blue sclerae and had the highest fracture frequency. Furthermore, there were some minor phenotypic differences among non-collagen I gene mutations. Interestingly, pamidronate achieved excellent results in the treatment of patients with OI, and the treatment effect appeared to be unrelated to their genotypes. CONCLUSION: Our findings indicated a genotype-phenotype relationship and a similar effect of pamidronate treatment in patients with OI, which could provide a basis for guiding clinical treatment and predicting OI prognosis.

Identification of proteins associated with bast fiber growth of ramie by differential proteomic analysis.

BACKGROUND: Ramie is an important fiber-producing crop in China, and its fibers are widely used as textile materials. Fibers contain specialized secondary cellular walls that are mainly composed of cellulose, hemicelluloses, and lignin. Understanding the mechanism underlying the secondary wall biosynthesis of fibers will benefit the improvement of fiber yield and quality in ramie. RESULTS: Here, we performed a proteomic analysis of the bark from the top and middle parts of the stem, where fiber growth is at different stages. We identified 6971 non-redundant proteins from bast bark. Proteomic comparison revealed 983 proteins with differential expression between the two bark types. Of these 983 proteins, 46 were identified as the homolog of known secondary wall biosynthetic proteins of Arabidopsis, indicating that they were potentially associated with fiber growth. Then, we proposed a molecular model for the secondary wall biosynthesis of ramie fiber. Furthermore, interaction analysis of 46 candidate proteins revealed two interacting networks that consisted of eight cellulose biosynthetic enzymes and seven lignin biosynthetic proteins, respectively. CONCLUSION: This study sheds light on the proteomic basis underlying bast fiber growth in ramie, and the identification of many candidates associated with fiber growth provides important basis for understanding the fiber growth in this crop.

Comparison of polypropylene mesh and porcine-derived, cross-linked urinary bladder matrix materials implanted in the rabbit vagina and abdomen.

INTRODUCTION AND HYPOTHESES: Our aim was to compare histological and biomechanical effects of polypropylene (PP) mesh and porcine-derived, cross-linked urinary bladder matrix (cUBM) graft materials using a rabbit vaginal and abdominal model. METHODS: Forty rabbits were implanted with PP mesh (n = 20) or cUBM (n = 20) in the vagina and abdomen. Two grafts (PP or cUBM) of the same type were placed into each site, so each rabbit had four grafts. Grafts were harvested 12 weeks later and processed for histologic analysis and biomechanical testing. RESULTS: There were high rates of two types of grafts missing in the vagina. Vaginal PP was associated with erosion reaction (67%), whereas abdominal PP and cUBM showed no sign of erosion. All patches adhered to rectus abdominis or vaginal mucosa and shrank to varying degrees, especially for PP grafts. Compared with vaginal PP, vaginal cUBM induced milder chronic inflammation response, had lower scores (P = 0.000) for inflammation response, and showed higher scores for neovascularization (P = 0.000) and fibroblastic proliferation (P = 0.002). In the abdomen, both histopathological parameters were insignificantly different (P > 0.05) between cUBM and PP. The mechanical properties of UBM did not deteriorate following implantation, whereas the ultimate tensile strength and elastic modulus of vaginal PP increased. PP had higher scores for tensile and break strength than did cUBM (P < 0.05). CONCLUSIONS: The cUBM has good biocompatibility, high ability to integrate with the vagina, and maintains mechanical properties in vivo. It may be a promising material for pelvic floor reconstruction.

Porcine urinary bladder matrix-polypropylene mesh: a novel scaffold material reduces immunorejection in rat pelvic surgery.

INTRODUCTION AND HYPOTHESIS: The present study set out to modify polypropylene vaginal surgical material using porcine urinary bladder matrix (UBM) in order to improve biocompatibility. The aim was to develop a compound scaffold that induced less vaginal erosion and to evaluate host immunoreactivity to this material in vivo. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into four equal groups. One group underwent a sham operation, and the other groups underwent vaginal implantation with different materials: UBM (U); UBM + polypropylene (UP); or polypropylene (P). The host tissue response was determined by macro-observation, and by histological and immunohistochemical methods at 7, 14, 21, or 28 days after surgery. RESULTS: The inflammation reaction was strongest throughout the entire observation time in Group P, but was weaker and had a tendency to decrease with time in Groups U and UP. The presence of the UBM material in the compound scaffold allowed the polypropylene to fuse with newly proliferating surrounding tissue and resulted in less rejection of the material by the host, as indicated by the reduced appearance of CD4-, and CD8-positive cells. CONCLUSIONS: Porcine UBM allowed mechanical isolation of polypropylene, and also reduced the immune reaction to polypropylene. This study suggests that the UBM + polypropylene compound scaffold may be a promising material for clinical use in pelvic reconstruction surgery.

Integration of Quantitative Trait Loci Mapping and Expression Profiling Analysis to Identify Genes Potentially Involved in Ramie Fiber Lignin Biosynthesis.

Ramie fibers, one of the most important natural fibers in China, are mainly composed of lignin, cellulose, and hemicellulose. As the high lignin content in the fibers results in a prickly texture, the lignin content is deemed to be an important trait of the fiber quality. In this study, the genetic basis of the fiber lignin content was evaluated, resulting in the identification of five quantitative trait loci (QTLs). Three genes, whole_GLEAN_10021050, whole_GLEAN_10026962, and whole_GLEAN_10009464 that were identified on the QTL regions of qLC7, qLC10, and qLC13, respectively, were found to be homologs of the Arabidopsis lignin biosynthetic genes. Moreover, all three genes displayed differential expression in the barks located in the top and middle parts of the stem, where lignin was not being synthesized and where it was being biosynthesized, respectively. Sequence comparison found that these three genes had wide variations in their coding sequences (CDSs) and putative promoter regions between the two parents, especially the MYB gene whole_GLEAN_10021050, whose protein had insertions/deletions of five amino acids and substitutions of two amino acids in the conserved domain. This evidence indicates that these three genes are potentially involved in lignin biosynthesis in ramie fibers. The QTLs identified from this study provide a basis for the improvement of lignin content and fiber quality in ramie breeding. The characterization of the three candidate genes here will be helpful for the future clarification of their functions in ramie.

Splice receptor-site mutation c.697-2A>G of the COL1A1 gene in a Chinese family with osteogenesis imperfecta.

Osteogenesis imperfecta (OI) is a genetic disorder characterized by bone fragility and blue sclerae, which are mainly caused by a mutation of the COL1A1 or COL1A2 genes that encode type I procollagen. Mutations in the splice site of type I collagen genes are one of the mutations that cause OI and usually lead to a mild or moderate OI phenotype. A heterozygous A to G point mutation in intron 9 at the -2 position of the splice receptor site of COL1A1 was identified in a family with type I or IV OI. Three affected individuals in four generations of one family all presented with several clinical symptoms. They all had pectus carinatum, flat feet, gray-blue sclerae, and normal stature, teeth, hearing, and vision. Forearm fractures, small joint dislocations, and muscle weakness were all present in the patient's father and grandmother, who presented with a moderate type IV phenotype. The 10-year-old proband with type I OI had suffered a fracture twice, but had no history of joint dislocation or skin hyperextensibility. Charting the family helped to identify clinical symptoms in patients with mutations at the N-terminal of type I collagen genes.

Heterozygous mutation of c.3521C>T in COL1A1 may cause mild osteogenesis imperfecta/Ehlers-Danlos syndrome in a Chinese family.

Osteogenesis imperfecta (OI) is an inheritable connective tissue disorder with a broad clinical heterozygosis, which can be complicated by other connective tissue disorders like Ehlers-Danlos syndrome (EDS). OI/EDS are rarely documented. Most OI/EDS mutations are located in the N-anchor region of type I procollagen and predominated by glycine substitution. We identified a c.3521C>T (p.A1174V) heterozygous mutation in COL1A1 gene in a four-generation pedigree with proposed mild OI/EDS phenotype. The affected individuals had blue sclera and dentinogenesis imperfecta (DI) was uniformly absent. The OI phenotype varied from mild to moderate, with the absence of scoliosis and increased skin extensibility. Easy bruising, joint dislocations and high Beighton score were present in some affected individuals. EDS phenotype is either mild or unremarkable in some individuals. The mutation is poorly conserved and in silico prediction support the relatively mild phenotype. The molecular mechanisms of the mutation that leads to the possible OI/EDS phenotype should be further identified by biochemical analysis of N-propeptide processing and steady state collagen analysis.

Histological response to and immunogenicity of different material patches implanted in rabbit abdominal walls.

The high failure rate of traditional procedures of repairing pelvic organ prolapse by using sutures pushes multiple transvaginal mesh delivery systems into the marketplace; however, these are currently without optimal clinical results. We observed the short-term histological changes and the effects of Th1/Th2 cytokines after the implantation in rabbit abdominal walls of five materials, i.e., porcine urinary bladder matrix (UBM), cross-linked UBM, polypropylene mesh, composite cross-linked UBM, and composite UBM, and explored the feasibility of their application to pelvic reconstructive surgery. Grafts were harvested at 1, 2, 4, 8, and 12 weeks after surgery and were processed for histology in order to evaluate the host inflammatory response and the degree of tissue incorporation. Additionally, graft site cytokine levels were determined by reverse transcriptase polymerase chain reaction. The results demonstrate that the two composite groups were associated with an erosion response, whereas the other groups induced a milder response than the composite groups. The Th1/Th2 cytokine mRNA expression levels of the cross-linked UBM group at each time point were similar to that of the sham group, whereas the other groups elicited a more variable expression. The cross-linked UBM showed slow degradation, mild inflammatory response, and low immunogenicity. This suggests that cross-linked UBM is a better material of choice for pelvic reconstructive surgery.