The Modified Rectangle Flap Epicanthoplasty: A Novel and Individualized Design.

BACKGROUND: The epicanthal fold is ordinary in the eyelids of Asians, and the aesthetic appearance of eyelid surgery could be reduced and undermined; thus, medial epicanthoplasty is commonly performed to eliminate the effect of the epicanthal fold with less scarring. At present, there are a lot of techniques that have been described for the treatment of epicanthal fold. The potential problems, however, such as visible scar or under correction in the medial canthus area are challenges to surgeons. The purpose of our study was to explore a novel and individualized design using a modified rectangle flap with acceptable functional and aesthetic outcomes. METHODS: From January 2017 to January 2018, epicanthoplasty was performed for 40 patients by using a modified rectangle flap. All patients underwent double-eyelid surgery at the same time when they needed it. The evaluation criteria included the intercanthal distance (ICD), interpupillary distance (IPD), the ratio of ICD to IPD (ICD ratio), scar visibility, and cosmetic results. RESULTS: From January 2017 to January 2018, the modified rectangle flap method was carried out on 40 patients, who were evaluated at follow-up from 7 to 15 months. The average intercanthal length was 36.9 ± 2.2 mm preoperatively and decreased significantly to 31.5 ± 1.8 mm postoperatively, 7 months after the surgery (P < 0.01). The excellent cosmetic results, in terms of an open medial canthus, were observed during follow-up periods, with no definite recurrence, hypertrophic scar, or injury of the lacrimal apparatus. The inner canthus and lacrimal caruncle are fully exposed with an invisible scar. Both the patients and the surgeon judged that the aesthetic outcomes were excellent or good. CONCLUSIONS: This modified rectangular flap is an effective and personalized method of correcting the medial folds that leave no additional scar in the medial canthal area, and the procedure meets the patient's aesthetic expectations. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

BMP2/7 heterodimer enhances osteogenic differentiation of rat BMSCs via ERK signaling compared with respective homodimers.

Bone morphogenetic protein (BMP)2/7 heterodimer shows greater efficacy in enhancing bone regeneration. However, the precise mechanism and the role of mitogen-activated protein kinase (MAPK) signaling network in BMP2/7-driven osteogenesis remain ambiguous. In this study, we evaluated the effects of BMP2/7 heterodimers on osteoblastic differentiation in rat bone marrow mesenchymal stem cells (BMSCs), with the aim to elaborate how MAPKs might be involved in this cellular process by treatment of rat BMSCs with BMP2/-7 with a special signal-pathway inhibitor. We found that BMP2/7 heterodimer induced a much stronger osteogenic response in rat BMSCs compared with either homodimer. Most interestingly, extracellular signal-regulated kinase (ERK) demonstrated a highly sustained phosphorylation and activation in the BMP2/7 heterodimer treatment groups, and inhibition of ERK cascades using U0126 special inhibitor that significantly reduced the activity of ALP and calcium mineralization to a substantial degree in rat BMSCs treated with BMP2/7 heterodimers. Collectively, we demonstrate that BMP2/7 heterodimer shows a potent ability to stimulate osteogenesis in rat BMSCs. The activated ERK signaling pathway involved in this process may contribute partially to an increased osteogenic potency of heterodimeric BMP2/7 growth factors.

Ultrastructural changes in human masseter muscles after botulinum neurotoxin a injection.

INTRODUCTION: Botulinum neurotoxin A (BoNTA) has long been used as a therapeutic agent and has been widely accepted as a cosmetic agent in recent years. It can inhibit function and induce structural changes in skeletal muscle. METHODS: Specimens of fresh dissected human masseter muscle were used to observe the ultrastructural changes that occurred at 6 and 12 months following BoNTA injection. RESULTS: The findings observed were muscle fiber distortion, sarcomere shortening, mitochondrial vacuolar degeneration, glycogen accumulation, and H and M band disruption in the triad of tubules. At 12 months after injection, there was still evidence of degenerative changes in muscle ultrastructure, whereas most organelles exhibited a normal structure. DISCUSSION: Profound ultrastructural and organelle disfiguring changes were observed after BoNTA injection into human masseter muscle. Most changes were transient, however, and were resolved by 12 months after injection. Muscle Nerve 57: 96-99, 2018.

PKM2 Thr454 phosphorylation increases its nuclear translocation and promotes xenograft tumor growth in A549 human lung cancer cells.

Pyruvate kinase M2 (PKM2) is a key enzyme of glycolysis which is highly expressed in many tumor cells, and plays an important role in the Warburg effect. In previous study, we found PIM2 phosphorylates PKM2 at Thr454 residue (Yu, etl 2013). However, the functions of PKM2 Thr454 modification in cancer cells still remain unclear. Here we find PKM2 translocates into the nucleus after Thr454 phosphorylation. Replacement of wild type PKM2 with a mutant (T454A) enhances mitochondrial respiration, decreases pentose phosphate pathway, and enhances chemosensitivity in A549 cells. In addition, the mutant (T454A) PKM2 reduces xenograft tumor growth in nude mice. These findings demonstrate that PKM2 T454 phosphorylation is a potential therapeutic target in lung cancer.

Zygomatic arch reduction and malarplasty with multiple osteotomies: its geometric considerations.

BACKGROUND: The midfacial width is dominated by the lateral protruding degree of the zygomatic arch. The best way of narrowing the midface is to reduce the arch height and the arc length for patients with an overly curved lateral protruding zygomatic arch. The existing techniques for reduction malarplasty cannot change the degree of curvature of the zygomatic arch. We provide a new technique for efficient midfacial width reduction by multiple osteotomies at different sites on the zygomatic complex and bone resection at the most protruding middle part of the zygomatic arch. The amount of bone resection can be calculated with a simplified geometrical solution according to the desired reduction rate of the arch height. METHODS: A digitalized CT image was used to estimate the arch height and the length of bone for removal from the zygomatic arch. A specific piece of bone was removed from the most protruding point of each zygomatic arch. Greenstick fractures were made at the anterior and posterior roots of the zygomatic arch. The open arches were rotated inwardly until both ends met. RESULT: The arch heights of 1,020 sides of the zygomatic arch were reduced in a range from 3 to 11 mm. All the reduced zygomatic arches were reunited properly and healed solidly. The overall satisfaction rate was high. CONCLUSION: This technique reduces the width of the midface by changing the degree of curvature of the zygomatic arch. The simplified geometrical calculation solutions are helpful in assuring the reunion of the zygomatic arch at a pre-designed lower arc height level after a calculated shortening of the arc length. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Highly-expressed lncRNA FOXD2-AS1 in adipose mesenchymal stem cell derived exosomes affects HaCaT cells via regulating miR-185-5p/ROCK2 axis.

The healing of skin wounds is a highly coordinated multi-step process that occurs after trauma including surgical incisions, thermal burns, and chronic ulcers. In this study, the authors investigated lncRNA FOXD2-AS1 function in adipose mesenchymal exosomes from ADMSCs that were successfully extracted. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes accelerated HaCaT cell migration and proliferation. LncRNA FOXD2-AS1 negatively targeted miR-185-5p, and miR-185-5p negatively targeted ROCK2. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes promoted HaCaT cell migration and proliferation via down-regulating miR-185-5p and further up-regulating ROCK2. In conclusion, LncRNA FOXD2-AS1 overexpression in ADMSCs derived exosomes might accelerate HaCaT cell migration and proliferation via modulating the miR-185-5p/ROCK2 axis.

MicroRNA-573 inhibits cell proliferation, migration and invasion and is downregulated by PICSAR in cutaneous squamous cell carcinoma.

The incidence of cutaneous squamous cell carcinoma (cSCC) has been increasing in recent years. Meanwhile, microRNAs (miRNAs) have been found to play vital roles in various cancers, including cSCC. This study aimed to investigate the expression of microRNA-573 (miR-573) in cSCC, its relationship with long non-coding RNA PICSAR and analyze its biological role. The relationship between PICSAR and miR-573 was confirmed by dual-luciferase reporter assay and Pearson's correlation coefficient analysis. The levels of PICSAR and miR-573 were measured using quantitative Real-Time PCR. Cell Counting Kit-8 assay was used to evaluate the cSCC cell proliferation ability. The migration and invasion abilities of cSCC cells were evaluated by Transwell assay. PICSAR expression was increased and miR-573 was decreased in tumor tissues and cSCC cell lines. PICSAR and miR-573 can bind directly, and miR-573 expression was downregulated by PICSAR in cSCC. Overexpression of miR-573 significantly inhibited the proliferation, migration and invasion abilities of A431 and SCC13 cells. Additionally, miR-573 overexpression reversed the promotion effects of PICSAR overexpression on cSCC cell proliferation, migration and invasion abilities. In conclusion, our findings indicated that miR-573 expression was decreased in tumor tissues and cSCC cells and was downregulated by PICSAR in cSCC. Additionally, miR-573 overexpression inhibited cSCC cell proliferation, migration and invasion, and reversed the promotion effects of PICSAR overexpression on cSCC cell biological functions. Thus, miR-573 might function as a tumor suppressor and might be involved in the regulatory effects of PICSAR on tumorigenesis in cSCC.

ITGB5 mutation discovered in a Chinese family with blepharophimosis-ptosis-epicanthus inversus syndrome.

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal-dominant genetic disorder, and mutations in the forkhead box L2 (FOXL2) gene are one of the major genetic causes. As this study shows, there are many patients with BPES who do not have FOXL2 mutations, as the screening results in all family members were negative. Using whole-exome sequence analysis, we discovered another possible mutational cause of BPES in integrin subunit beta 5 (ITGB5). The ITGB5 mutation (c.608T>C, p.Ile203Thr) appears in the base sequence of all BPES+ patients in this family, and it appears to be a three-generation-inherited mutation. It can cause changes in base sequence and protein function, and there may be cosegregation of disease phenotypes. ITGB5 is located on the long arm of chromosome three (3q21.2) and is close to the known pathogenic gene FOXL2 (3q23). This study is the first to report ITGB5 mutations in BPES, and we speculate that it may be directly involved in the pathogenesis of BPES or indirectly through the regulation of FOXL2.

Ultrafast, tough, and adhesive hydrogel based on hybrid photocrosslinking for articular cartilage repair in water-filled arthroscopy.

A hydrogel scaffold for direct tissue-engineering application in water-irrigated, arthroscopic cartilage repair, is badly needed. However, such hydrogels must cure quickly under water, bind strongly and permanently to the surrounding tissue, and maintain sufficient mechanical strength to withstand the hydraulic pressure of arthroscopic irrigation (~10 kilopascal). To address these challenges, we report a versatile hybrid photocrosslinkable (HPC) hydrogel fabricated though a combination of photoinitiated radical polymerization and photoinduced imine cross-linking. The ultrafast gelation, high mechanical strength, and strong adhesion to native tissue enable the direct use of these hydrogels in irrigated arthroscopic treatments. We demonstrate, through in vivo articular cartilage defect repair in the weight-bearing regions of swine models, that the HPC hydrogel can serve as an arthroscopic autologous chondrocyte implantation scaffold for long-term cartilage regeneration, integration, and reconstruction of articular function.

Adipose stem cell secretion combined with biomaterials facilitates large-area wound healing.

Adipose-derived stem cell (ADSC)-based therapeutic strategies are in fast-pace advancement in wound treatment due to their availability and the ability to self-renew, undergo multilineage differentiation and self-renewal. Existing studies have successfully explored ADSCs to facilitate scar-free healing of small wounds, but whether the healing of large-area wounds that exhibit over 50% of skin tissue loss in the entire body could be achieved remains controversial. This study sought to review the mechanism of physiological wound healing, and discuss the roles played by chemokines, biological factors and biomaterial scaffolds. The possibility of applying ADSC-conditioned medium or ADSC-released exosomes as 'off-the-shelf' tissue engineering products, integrated with biomaterial scaffolds to facilitate wound healing, was analyzed.

Effects of chondrogenic microenvironment on construction of cartilage tissues using marrow stromal cells in vitro.

OBJECTIVE: To investigate whether it is feasible to use the chondrogenic microenvironment provided by cartilage cells to construct cartilage tissues in vitro with bone marrow stromal cells (BMSC). MATERIALS AND METHODS: We isolated and cultured BMSC and cartilage cells from Sprague Dawley rats (SD rats). The supernatant of cartilage culture was used as inducing solution to cause differentiation of BMSC from the second generation of cells cultured in vitro. Cells were examined seven days later, using immunohistochemistry to determine the expression of collagen specific to type II cartilage. RT-PCR was used to detect the expression of type II collagen and aggrecan mRNA. BMSC and cartilage cells were isolated from SD rats and cultured in vitro. The BMSC and cartilage cells in culture were mixed evenly in an 8:2 ratio and inoculated into a polyglycolic acid/polylactic acid (PGA/PLA) scaffold to a final concentration of 5.0x10(7) cells/ml. PGA/PLA preparations with pure cartilage cells or pure BMSC served as the positive and negative controls, respectively. The control group of low-concentration cartilage cells consisted of PGA/PLA preparations containing cartilage cells at 20% of the above mentioned concentration (1.0x10(7) cells/ml). Samples were collected eight weeks later, at which time general observations, wet weight, and glycosaminoglycan (GAG) levels were determined, and histological and immunohistochemical examinations were performed. RESULTS: Immunohistochemistry showed the induction of BMSC type II collagen, and RT-PCR indicated the expression of type II collagen and aggrecan mRNA. In the mixed-cell group and the positive control group, pure mature cartilage cells were produced after eight weeks of culture in vitro, and the size and shape of the scaffold were maintained throughout the culture period. The two groups gave rise to newly generated cartilage cells essentially identical in appearance and histological properties. The immunohistochemical results showed that the cartilage cells of both groups expressed abundant cartilage-specific type II collagen. The average wet weight and GAG content were more than 70% of the values in the positive control group. Only an extremely small amount of immature cartilage tissue formed in local regions in the BMSC-only sample, and the scaffold was obviously shrunken and deformed. Although the wet weight of newly generated cartilage tissue in the low-concentration cartilage cell sample reached 30% of the value of the positive control group, the scaffold was obviously shrunken and deformed. Only regional and discontinuous cartilage tissues were formed, and the amount of newly generated cartilage was obviously less than in the co-culture and positive control groups. CONCLUSIONS: Cartilage cells can provide a microenvironment for cartilage formation to some extent, and also effectively induce BMSC to differentiate into cartilage cells and form tissue-engineered cartilage in vitro.

[Analysis of risk factors related to periprosthetic infection after breast augmentation].

OBJECTIVE: To explore the risk factors related to periprosthetic infection after breast augmentation, and to provide a basis for reducing the risk of postoperative infection. METHODS: A total of 1 056 female patients who underwent breast augmentation between January 2010 and January 2018 were analyzed retrospectively. The patients were 20 to 44 years old (mean, 31.6 years). The body mass index (BMI) was 19.0-31.1 kg/m 2, with an average of 24.47 kg/m 2. According to the periprosthetic infection standard of the United States Centers for Disease Control and Prevention (CDC), the patients were divided into infection group and non-infection group. Age, BMI, diabetes, previous history of immunosuppression, history of smoking, previous history of breast surgery, previous history of mastitis, combined with active dermatitis, surgical approach, the type and shape of breast prosthesis, implant in the different layers, combined with mastopexy, operation time, postoperative antibiotic time, postoperative breast crash, and postoperative potential infection surgery were analyzed by univariate analysis. The influencing factors of prosthetic infection were screened by logistic regression. RESULTS: Periprosthetic infection occurred in 60 cases after operation, and the infection rate was 5.68%. Among them, 11 cases were acute infection, 33 cases were subacute infection, 16 cases were delayed infection, and 20 cases were positive in bacterial culture. Postoperative breast crash occurred in 114 cases. Univariate analysis showed that diabetes, previous history of immunosuppression, history of smoking, previous history of mastitis, postoperative breast crash, postoperative potential infection surgery, and combined with breast suspension were the influencing factors of postoperative periprosthetic infection ( P<0.05). Multivariate analysis showed that diabetes, history of smoking, and postoperative breast crash were the risk factors of periprosthetic infection ( P<0.05). CONCLUSION: Diabetes, smoking, and postoperative breast crash are the risk factors of periprosthetic infection after breast augmentation.

[Effectiveness of levator muscle resection combined with Mustarde's double Z-plasty for blepharophimosis-ptosis-epicanthus inversus syndrome].

Objective: To investigate the effectiveness of levator muscle resection combined with Mustarde's double Z-plasty to correct blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). Methods: Between March 2015 and June 2017, one-stage operation of levator muscle resection combined with Mustarde's double Z-plasty were performed on 26 children with bilateral BPES. There were 16 boys and 10 girls with an average age of 7 years (range, 4-14 years). All patients marked the four typical signs of BPES. There were 7 cases accompanied with a low nasal bridge, and 20 cases with amblyopia and strabismus. The length of eye fissure was (19.5±4.5) mm, the width of eye fissure was (2.5±1.6) mm, the diameter of inner canthus was (42.1±6.5) mm, and the muscular strength of levator palpebrae superioris was (5.5±1.3) mm. Results: All the incisions healed by first intention. Twenty-three patients were followed up 2-12 months, with an average of 10 months. Among which, 2 cases were less corrected, 3 cases were over corrected, 6 cases had poor curvature of the eyelid. No eyelid internal and external pronation or keratitis occurred. Amelioration of blepharoptosis and epicanthus was achieved in the other patients, and the double eyelid fold was naturally smooth. At 7 days after operation, the length of eye fissure was (27.2±1.9) mm, the width of eye fissure was (12.5±1.3) mm, and diameter of inner canthus was (29.4±2.6) mm, which were superior to preoperative values ( t=0.127, P=0.042; t=0.341, P=0.029; t=0.258, P=0.038). There was no angular deformity caused by the width and length regressions of eye fissures. Conclusion: The levator muscle resection combined with Mustarde's double Z-plasty can effectively correct BPES and obtain good effectiveness.

Cell yield, chondrogenic potential, and regenerated cartilage type of chondrocytes derived from ear, nasoseptal, and costal cartilage.

Functional reconstruction of large cartilage defects in subcutaneous sites remains clinically challenging because of limited donor cartilage. Tissue engineering is a promising and widely accepted strategy for cartilage regeneration. To date, however, this strategy has not achieved a significant breakthrough in clinical translation owing to a lack of detailed preclinical data on cell yield and functionality of clinically applicable chondrocytes. To address this issue, the current study investigated the initial cell yield, proliferative potential, chondrogenic capacity, and regenerated cartilage type of human chondrocytes derived from auricular, nasoseptal, and costal cartilage using a scaffold-free cartilage regeneration model (cartilage sheet). Chondrocytes from all sources exhibited high sensitivity to basic fibroblast growth factor within 8 passages. Nasoseptal chondrocytes presented the strongest proliferation rate, whereas auricular chondrocytes obtained the highest total cell amount using comparable cartilage sample weights. Importantly, all chondrocytes at fifth passage showed strong chondrogenic capacity both in vitro and in the subcutaneous environment of nude mice. Although some significant differences in histological structure, cartilage matrix content and cartilage type specific proteins were observed between the in vitro engineered cartilage and original tissue; the in vivo regenerated cartilage showed mature cartilage features with high similarity to their original native tissue, except for minor matrix changes influenced by the in vivo environment. The current study provides detailed preclinical data for choice of chondrocyte source and thus promotes the clinical translation of cartilage regeneration approach.

Transfection of adenovirus-mediated mircoRNA-126 gene into infant hemangioma endothelial cells in vitro.

Objective: To investigate the effects of microRNA-126 (miR-126) overexpression on hemangioma endothelial cells (HemECs). Methods: An adenoviral vector containing the miR-126 gene was constructed. HemECs were passaged and expanded and adenovirus-mediated green fluorescent protein (GFP) gene was transfected in vitro. The infection efficiency of adenovirus vector to HemECs was tested by Ad-GFP infection procedure. GFP expression efficiency was observed using a fluorescence microscope and flow cytometry was used to determine the best virus multiplicity of infection (MOI). The experiment was divided into the blank group, AD-GFP group, and AD-miR-126 group. The miR-126 group was transfected into HemECs in vitro with adenovirus-mediated miR-126 gene under optimal MOI conditions. RT-PCR was applied to detect expression of miR-126 gene in cells. The influence of recombinant adenovirus on cell activity was evaluated by CCK-8 assay. Flow cytometry was utilized to detect cell cycle and apoptosis. Results: HemECs could be effectively infected by adenovirus containing GFP gene in vitro, the transfection efficiency had the dose-effect relationship with multiplicities of infection (MOI). When MOI was 400, the infection efficiency was more than 90%. miR-126 expression in HemECs was significantly enhanced in miR-126 group (P<0.05). Compared to the control group, cell proliferation was significantly enhanced (P<0.05) and induced S-phase arrest significantly (P<0.05) when miR-126 was upregulated. In addition, compared with the control group, the early apoptotic rate was significantly decreased by upregulating miR-126 (P<0.05). Conclusion: miR-126 overexpression can successfully promote proliferation and inhibit apoptosis of HemECs. This work will provide the theoretical and experimental basis for further transplantation study in vivo.

Lyophilized Scaffolds Fabricated from 3D-Printed Photocurable Natural Hydrogel for Cartilage Regeneration.

Repair of cartilage defects is highly challenging in clinical treatment. Tissue engineering provides a promising approach for cartilage regeneration and repair. As a core component of tissue engineering, scaffolds have a crucial influence on cartilage regeneration, especially in immunocompetent large animal and human. Native polymers, such as gelatin and hyaluronic acid, have known as ideal biomimetic scaffold sources for cartilage regeneration. However, how to precisely control their structure, degradation rate, and mechanical properties suitable for cartilage regeneration remains a great challenge. To address these issues, a series of strategies were introduced in the current study to optimize the scaffold fabrication. First, gelatin and hyaluronic acid were prepared into a hydrogel and 3D printing was adopted to ensure precise control in both the outer 3D shape and internal pore structure. Second, methacrylic anhydride and a photoinitiator were introduced into the hydrogel system to make the material photocurable during 3D printing. Finally, lyophilization was used to further enhance mechanical properties and prolong degradation time. According to the current results, by integrating photocuring 3D printing and lyophilization techniques, gelatin and hyaluronic acid were successfully fabricated into human ear- and nose-shaped scaffolds, and both scaffolds achieved shape similarity levels over 90% compared with the original digital models. The scaffolds with 50% infill density achieved proper internal pore structure suitable for cell distribution, adhesion, and proliferation. Besides, lyophilization further enhanced mechanical strength of the 3D-printed hydrogel and slowed its degradation rate matching to cartilage regeneration. Most importantly, the scaffolds combined with chondrocytes successfully regenerated mature cartilage with typical lacunae structure and cartilage-specific extracellular matrixes both in vitro and in the autologous goat model. The current study established novel scaffold-fabricated strategies for native polymers and provided a novel natural 3D scaffold with satisfactory outer shape, pore structure, mechanical strength, degradation rate, and weak immunogenicity for cartilage regeneration.

[Mutation analysis of FOXL2 gene and its structure protein in patients of blepharophimosis-ptosis-epicanthus inversus syndrome].

OBJECTIVE: The aim of this study was to investigate the effect of mutations of the forkhead transcription factor 2 (FOXL2) gene on the primary and secondary structure of the coded protein and seek for the molecular mechanism of blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). METHODS: The genomic DNA was extracted from peripheral blood of 7 clinically diagnosed BPES patients, PCR amplification of FOXL2 coding region and 5' untranslated region were performed. Sequence analysis was performed using the PCR or cloning products. The structure of the protein was predicted with PDH and ExPASy software, and the difference between the normal and the mutational protein was analyzed. RESULTS: A 901- 930 dup 30 mutation of FOXL2 was found in two patients from a BPES family of type II and a sporadic case, and no any mutations were detected in normal control. Analysis of the primary structure displayed that the molecular weight of the protein coded by the mutated gene was greater than the normal, but both have the same isoelectric point. Analysis of the secondary structure showed that FOXL2 was a transmembrane protein with a polyalanine tract which contained a alpha-helix. When the polyalanine tract expanded, the helix region extended, as a result, the proportion of alpha-helix increased by 4.1%, but the proportions of beta-pleated sheet and random coil decreased correspondingly. CONCLUSION: Our results suggest that the 901 - 930 dup 30 mutation of FOXL2 is a novel finding. Moreover, this mutation causes great changes in the primary and secondary structure of the coded protein, which may be the molecular pathogenesis of BPES.

Cloning of the promoter region of a human gene, FOXL2, and its regulation by STAT3.

Forkhead box L2 (FOXL2) is a transcription factor, which is involved in blepharophimosis, ptosis, and epicanthus in versus syndrome (BPES), premature ovarian failure (POF), as well as almost all stages of ovarian development and function. FOXL2 has various target genes, which are implicated in numerous processes, including sex determination, cell cycle regulation and apoptosis and stress response regulation in mammals. However, studies regarding the upstream regulation of FOXL2 are limited. In the present study, the promoter of FOXL2 was successfully cloned and registered in Gen Bank, and a dual luciferase reporter (DLR) analysis demonstrated that the luciferase activity was significantly induced by the promoter of FOXL2. Subsequently, bioinformatics analysis indicated that FOXL2 may be regulated by STAT3, and this was confirmed by a DLR analysis and western blotting, using STAT3 inhibitors. Further study using real­time cellular analysis indicated that the viability of He La cells was markedly suppressed by STAT3 inhibitors. The present study demonstrated novel findings regarding the upstream regulation of FOXL2 expression and provide a new perspective for future studies in the field.

Osteogenic Differentiation Capacity of In Vitro Cultured Human Skeletal Muscle for Expedited Bone Tissue Engineering.

Expedited bone tissue engineering employs the biological stimuli to harness the intrinsic regenerative potential of skeletal muscle to trigger the reparative process in situ to improve or replace biological functions. When genetically modified with adenovirus mediated BMP2 gene transfer, muscle biopsies from animals have demonstrated success in regenerating bone within rat bony defects. However, it is uncertain whether the human adult skeletal muscle displays an osteogenic potential in vitro when a suitable biological trigger is applied. In present study, human skeletal muscle cultured in a standard osteogenic medium supplemented with dexamethasone demonstrated significant increase in alkaline phosphatase activity approximately 24-fold over control at 2-week time point. More interestingly, measurement of mRNA levels revealed the dramatic results for osteoblast transcripts of alkaline phosphatase, bone sialoproteins, transcription factor CBFA1, collagen type I, and osteocalcin. Calcified mineral deposits were demonstrated on superficial layers of muscle discs after an extended 8-week osteogenic induction. Taken together, these are the first data supporting human skeletal muscle tissue as a promising potential target for expedited bone regeneration, which of the technologies is a valuable method for tissue repair, being not only effective but also inexpensive and clinically expeditious.