Résumé
Abstract Background The development of rosacea is suggested to be closely associated with lipid metabolism, inflammation, and anxiety/depression. Gamma linolenic acid (GLA) is a key factor participating in lipid metabolism, which is also confirmed to regulate the inflammatory response. However, the associations of serum GLA levels with rosacea severity and psychological status still remain unclear. Objective and limitations of the study The present study aimed to investigate the associations of gamma linolenic acid (GLA), a key factor participating in lipid metabolism and the inflammatory response, with rosacea severity and psychological status. The present study still had some limitations. First, this study is a cross-sectional study and does not provide longitudinal evidence about the relationship between GLA and rosacea; Second, the cohort in this study is also relatively small, and a larger cohort is needed in further investigation to reveal the potential role of lipid metabolism in the pathogenesis of rosacea. Methods A total of 62 rosacea patients were consecutively recruited. Patient's Self-Assessment (PSA) scale and Clinician Erythema Assessment (CEA) as well as 7-item Generalized Anxiety Disorder (GAD-7) and 9-item Patient Health Questionnaire (PHQ-9) were conducted to evaluate the degree of erythema severity and anxiety/depression, respectively. Serum GLA levels were determined by gas chromatography mass. Results Lower levels of serum GLA in rosacea patients were observed (p < 0.001), and subgroup analysis revealed that patients with higher-level GLA had lower scores of PSA, CEA, GAD-7 and PHQ-9. Moreover, Spearman correlation analysis uncovered that serum GLA levels were negatively associated with PSA, CEA, GAD-7 as well and PHQ-9 scores, respectively. Linear regression model found that serum GLA levels at baseline were a predictive factor for prognosis of clinical outcomes after 1-month conventional treatment. Conclusion The present study indicates that lower levels of serum GLA in rosacea patients are negatively associated with the degree of erythema and anxiety/depression status.
Résumé
The purpose of this study was to evaluate the effect of bone marrow mesenchymal stem cells(MSCs)transfected with the basic fibroblast growth factor(bFGF)-expressing recombinant adeno-associated virus vector(rAAV2-bFGF),on early angiogenesis of calvarial defects in rats.The MSCs were cultured and transfected with rAAV2-bFGF after differential adherence isolation.The transfection efficiency was detected by RT-PCR and Western blotting.The transfected MSCs were compounded with poly-DL-lactide/hydroxyapatite(PDLLA/HA)in vitro.The cranial defect models in36 malc SD rats were created.Nothing(group A),PDLLA/HA alone(group B),PDLLA/HA combined with MSCs(group C),and PDLLA/HA combined with rAAV2-bFGF transfected MSCs(group D)were implanted in rat calvarial defects.The specimens were harvested for hematoxylin-eosin staining on the day 1,3 and 7 after implantation.Factor Ⅷ immunohistochemical staining and histomorphometric analysis were carried out to evaluate neovascularization around the implantation.The results indicated that MSCs could indeed be successfully transfected with the rAAV2-bFGF vector.Histological and histomorphometric analysis revealed that the angiogenesis in group D was significantly enhanced as compared with the rest groups(P<0.05).These results strongly suggest that MSCs transfected with rAAV2-bFGF in combination with PDLLA/HA can effectively promote the early angiogenesis of calvarial defects in rats,which played an important role in creating an environment suitable for the survival and activity of transplanted cells for further applications in cranio-maxillofacial bone regeneration.
Résumé
To construct the recombinant adeno-associated virus (rAAV) vector with human bone morphogenetic protein 7 (BMP7) and observe the BMP7 mRNA expression in vitro, BMP7 CDS se- quence was cloned into expression plasmid pAAV-MCS of AAV Helper Free System. The recombi- nant plasmid was identified with enzyme digestion and sequencing. The recombinant plasmid, pAAV-RC, pHelper were co-transfected into AAV-293 cells according to the calcium phosphate-based protocol. The viral stock was collected by 4 rounds of freeze/thaw. After purified and concentrated,the recombinant virus titer was determined by dot-blot assay. HEK293 cells were transfected with the recombinant virus at different MOI, and the expression of BMP7 mRNA was detected by RT-PCR. The results showed rAAV-BMP7 was constructed and packaged successfully. The physical particle titer was 2.5×1011 vector genomes/mL. There was different expression level of BMP7 mRNA after transfecton. These data suggested that recombinant AAV mediated a stable expression of hBMP7 mRNA in 293 cells. The AAV production method may pave the way of an effective strategy for the jaw bone defection around dental implants.