Effect of Dichotomous Conchal Cartilage Transplantation on Correction of Unilateral Cleft Lip Nasal Deformity.

BACKGROUND: Autologous conchal cartilage is becoming increasingly popular as a source of material for secondary reconstruction. The aim of this study was to verify the effect of dichotomous autologous conchal cartilage transplantation in the treatment of unilateral cleft lip nasal deformity. METHODS: Eighteen patients (10 males and 8 females) with unilateral cleft lip nasal deformity treated from August 2018 to August 2019 were selected for the study. The cut C-shaped conchal cartilage was trimmed into a strip shape and a shield shape and transplanted into the alar cartilage and the tip of the nose, respectively. The effect of the operation was evaluated in terms of patient satisfaction, two-dimensional linear quantitative results, and three-dimensional spatial differences after the operation. RESULTS: During follow-up from 6 months to 2 years, the nasal appearance of 18 patients was significantly improved. The postoperative patient satisfaction survey revealed more than 93% satisfaction for each research index. Two-dimensional linear quantitative analysis revealed that the height of the nasal columella and nostril was significantly increased and that the nasal base and breadth were significantly decreased after the operation. Evaluation of the three-dimensional spatial difference between the unaffected side and the affected side before and after the operation revealed a significant decrease in the difference in the soft tissue volume between the unaffected side and the affected side (P < 0.05). CONCLUSIONS: Dichotomous autologous conchal cartilage transplantation is an ideal method for the treatment of unilateral cleft lip nasal deformity.

Downregulation of miR-10a inhibits cutaneous squamous cell carcinoma cell proliferation, migration, and invasion by targeting Syndecan-1.

BACKGROUND: Cutaneous squamous cell carcinoma (cSCC) is a malignancy of epidermal keratinocytes which accounts for approximately one-third of skin cancer-related death yearly. In this study, we aim to investigate the mechanism of miR-10a in regulating cellular function of cSCC cells and its possible role in prognosis of cSCC. METHODS: The expression of miR-10a was detected by qRT-PCR. Target mRNA candidates were detected by bioinformatic analysis. Proliferative and migration capability of cSCC cell were examined by MTT assay, wound healing assay, and invasion assay, respectively. miR-10a expression was monitored in cSCC patients to elucidate the relationship between miR-10a expression and outcomes of cSCC. RESULTS: In our study, we found that expression of miR-10a was significantly down-regulated in cSCC cell in vitro and in vivo. Moreover, our results revealed that SDC-1 was a likely target of miR-10a in regulating biologic function of cSCC cell. Additionally, miR-10a expression level was inversely correlated with cSCC cell differentiation and tumor progression. CONCLUSION: These findings in this study indicate the importance of miR-10a in cSCC cell hallmarks and its use as a novel target for cSCC treatment.

The cleft palate candidate gene BAG6 supports FoxO1 acetylation to promote FasL-mediated apoptosis during palate fusion.

BACKGROUND: Cleft palate is a common craniofacial defect, which occurs when the palate fails to fuse during development. During fusion, the palatal shelves migrate towards the embryonic midline to form a seam. Apoptotic elimination of medial edge epithelium (MEE) cells along this seam is required for the completion of palate fusion. METHODS: Whole exome sequencing (WES) of six Chinese cleft palate families was applied to identify novel cleft palate-associated gene variants. Palatal fusion and immunofluorescence studies were performed in a murine palatal shelf organ culture model. Gene and protein expression were analyzed by qPCR and immunoblotting in murine MEE cells during seam formation in vivo. Mechanistic immunoprecipitation studies were performed in murine MEE cells in vitro. RESULTS: WES identified Bcl-2 associated anthanogene 6 (BAG6) as a novel cleft palate-associated gene. In murine MEE cells, we discovered upregulation of Bag6 and the transcription factor forkhead box protein O1 (FoxO1) during seam formation in vivo. Using a palatal shelf organ culture model, we demonstrate that nuclear-localized Bag6 enhances MEE cell apoptosis by promoting p300's acetylation of FoxO1, thereby promoting transcription of the pro-apoptotic Fas ligand (FasL). Subsequent gain- and loss-of-function studies in the organ culture model demonstrated that FasL is required for Bag6/acFoxO1-mediated activation of pro-apoptotic Bax/caspase-3 signaling, MEE apoptosis, and palate fusion. Palatal shelf contact was shown to enhance Bag6 nuclear localization and upregulate nuclear acFoxO1 in MEE cells. CONCLUSIONS: These findings demonstrate that nuclear-localized Bag6 and p300 co-operatively enhance FoxO1 acetylation to promote FasL-mediated MEE apoptosis during palate fusion.

Long non-coding RNA PANDAR promotes melanoma cell invasion through regulating epithelial-mesenchymal transition.

Melanoma is a malignant skin tumor and has a poor cure rate because of its high metastatic potential. Overexpression of long non-coding (lnc) RNA PANDAR has been observed in several kinds of cancer, but the function of PANDAR on melanoma is still unclear. Therefore, this study was to explore the mechanism of PANDAR on the occurrence and progression in malignant melanoma. We detected expression of PANDAR in malignant melanoma tissues and cell lines by qRT-PCR and analyzed correlation of PANDAR expression with the patients' prognosis. Furthermore, we investigated the effects of PANDAR on cell viability, migration, invasion, tumorigenesis, and epithelial-mesenchymal transition (EMT) using CCK-8, Transwell, and nude mouse subcutaneous tumor formation model assays and Western blotting analysis, respectively. From the results, we discovered that the PANDAR expression is strikingly upregulated in melanoma tissues compared with paired-adjacent non-tumorous tissues and elevated PANDAR is positively correlated with short overall survival time. The results also demonstrate that knockdown of PANDAR inhibits cell viability, migration, invasion, tumorigenesis, and EMT, whereas overexpression of PANDAR gave opposite results by promoting cell viability, migration, invasion, tumorigenesis, and EMT of melanoma cells. These new findings all illustrate that PANDAR might play a pivotal oncogenic role in the occurrence and development of melanoma, and PANDAR might promote melanoma cell invasion through regulating EMT, providing a potential diagnostic and therapeutic target for melanoma.

Fibroproliferative effect of microRNA-21 in hypertrophic scar derived fibroblasts.

Hypertrophic scar (HS) is a fibroproliferative disorder caused by abnormal wound healing, which is characterized by excessive deposition of extracellular matrix (ECM) secreted by fibroblasts. We previous have found that expression of microRNA-21(miR-21) was increased in tissues and fibroblasts of HS. However, the underlying molecular mechanism remains to be further elucidated. In this study, we identified the miR-21 was a marker for the phenotype of HS fibroblasts, as anti-miR-21 reduced expression of fibrosis markers such as Col1A1, Col3A1, Fn and α-SMA in fibroblasts and overexpression of miR-21 promoted fibroproliferative expression in fibroblasts. Furthermore, we also found that miR-21 promoted TGF-ß1 induced fibroproliferative expression by repressing Smad7 expression in vitro. In addition, the miR-21 inhibitor inhibited the growth of hypertrophic scar tissue in vivo (nude mice experimental model). These results indicated that miR-21 was a critical regulator for HS formation and TGF- ß1/miR-21/Smad7 pathway could be a useful therapeutic target for the treatment of HS.

Bone regeneration in a rabbit ulna defect model: use of allogeneic adipose-derivedstem cells with low immunogenicity.

Tissue engineering provides new potential treatments for the repair of bone defects. Bone-marrow-derived mesenchymal stem cells (BMSCs) represent an attractive cell source for therapeutic applications involving tissue engineering, although disadvantages, such as pain of harvest and low proliferation efficiency, are major limitations to the application of BMSCs in the clinic. Adipose-derived stem cells (ASCs) with their multilineage potential and satisfactory proliferation potential can be induced into the osteogenic lineage in vitro and can be anchored onto suitable scaffolds as seed cells to repair bone defects successfully in an autologous setting. Previous studies have indicated that both undifferentiated BMSCs and ASCs exhibit immunosuppression and immunoprivilege properties. We compare the immuno-function of undifferentiated and osteo-differentiated ASCs in vitro and explore the feasibility of applying allogeneic ASCs to the repair of ulnar bone defects in the rabbit model. Our study demonstrates that allogeneic osteogenic differentiated ASCs maintain low immunogenicity and negative immunomodulation. The allogeneic osteogenic differentiated ASCs combined with demineralized bone matrix successfully regenerate ulnar bone defects in rabbits without immunosuppressive therapies.

Local injection of deferoxamine improves neovascularization in ischemic diabetic random flap by increasing HIF-1α and VEGF expression.

BACKGROUND: Although the systemic administration of deferoxamine (DFO) is protective in experimental models of normal ischemic flap and diabetic wound, its effect on diabetic flap ischemia using a local injection remains unknown. OBJECTIVE: To explore the feasibility of local injection of DFO to improve the survival of ischemic random skin flaps in streptozotocin (STZ)-induced diabetic mice. METHODS: Ischemic random skin flaps were made in 125 mice. Animals were divided into the DFO-treated (n = 20), PBS-treated (n = 16) and untreated (n = 16) groups. Surviving area, vessel density, and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) were evaluated on the seventh day after local injection. RESULTS: The viability of DFO-treated flap was significantly enhanced, with increased regional blood perfusion and capillary density compared with those in the two control groups. Fluorescence-activated cell sorting (FACS) analysis demonstrated a marked increase in systemic Flk-1+/CD11b- endothelial progenitor cells (EPCs) in DFO-treated mice. Furthermore, the expression of VEGF and HIF-1α was increased not only in diabetic flap tissue, but also in dermal fibroblasts cultured under hyperglycemic and hypoxic conditions. CONCLUSIONS: Local injection of DFO could exert preventive effects against skin flap necrosis in STZ-induced diabetic mice by elevating the expression of HIF-1α and VEGF, increased EPC mobilization, which all contributed to promote ischemic diabetic flap survival.

Effects of human adipose-derived stem cells on the viability of rabbit random pattern flaps.

BACKGROUND AIMS: Flap necrosis is the most commonly encountered outcome influencing the effect of operations in clinical practice. The advent of cytotherapy and regenerative medicine with stem cells, especially adipose-derived stem cell therapy, appears to be a promising approach in providing multi-lineage differentiating cells. However, autologous stem cells are limited in both quantity and quality in aging individuals. Hence, xenogenic stem cell therapy was used in this study. METHODS: Random pattern flaps (6 cm × 2 cm) were prepared in a rabbit model transplanted either with 4 × 10(5) human adipose-derived stem cells at five sites or equal volumes of Dulbecco's modified Eagle's medium. At 7 days after operation, the viability of the flaps from both groups was evaluated. We determined the numbers of locally infiltrating T cells, whereas the CD4/CD8 ratio, interferon, interleukin (IL)-2, IL-4 and IL-10 in the serum were determined to evaluate the immunological response of the rabbit. Moreover, Dil labeling was administrated to trace the homing of the transplanted stem cells. RESULTS: Both the survival areas and the capillary number of the flaps that were injected with human adipose-derived stem cells significantly increased as compared with the control group (P < 0.05). Additionally, no significant difference in the immune response was detected between the groups. Dil-labeled stem cells were found to participate in the formation of tubular structures, which were further shown to be CD31+, although not predominantly. CONCLUSIONS: Human adipose-derived stem cells could be used therapeutically to improve the viability of random pattern flaps without detection of serious immune rejection of stem cells.

The distally based lateral sural neuro-lesser saphenous veno-fasciocutaneous flap: anatomical basis and clinical applications.

BACKGROUND: Soft tissue management around the lower third of the leg and foot presents a considerable challenge to the plastic surgeon. The aim of this research was to investigate the anatomical relationships of artery, nerve, vein and other adjacent structures in the posterolateral region of the calf, and our experience with using a distally based island flap pedicled with the lateral sural nerve and the lesser saphenous vein for soft tissue reconstruction of lower third of leg, foot, and ankle defects in 15 patients. MATERIALS AND METHODS: Five fresh cadavers (ten lower limbs) were infused with colored red latex. The origin of the nutrient vessel of the lesser saphenous vein and the lateral sural nerve was identified. Based on the anatomical studies, an island flap supplied by the vascular axis of the lesser saphenous vein and the lateral sural nerve was designed for clinical reparative applications in 15 cases. RESULTS: The nutrient vessel of the lesser saphenous vein and the lateral sural nerve originates from the superficial sural artery, musculocutaneous perforators of the posterior tibial artery, and septocutaneous perforators of the peroneal artery in different segment of the calf. Meanwhile, these vessels have many sub-branches nourishing subcutaneous tissue and skin, form a favorable vascular chain around the nerve and the vein, and also communicate with vascular plexus of superficial and deep fascia. Among 15 flaps, 13 showed complete survival (86.66 %), while marginal flap necrosis occurred in one patient (6.67 %) and distal wound dehiscence in another (6.67 %). Their appearance and function were satisfactory, with feeling maintained in the heel and lateral side of the foot. CONCLUSIONS: The distally based flap pedicled with the lateral sural nerve and lesser saphenous vein was a reliable source for repairing soft tissue defects in the lower leg and foot due to its advantages of infection control, high survival rate, and sufficient blood supply without the need to sacrifice a major blood vessel.

Clinical and anatomical study of the distally based lesser saphenous veno-lateral sural neurocutaneous flap for lower extremity coverage.

BACKGROUND: The distally based sural flap has been widely and successfully used to reconstruct soft tissue defects of the distal third of the lower leg and foot. Sensory loss and venous congestion are possible complications of this treatment, but there has been limited research focused on improving the sensory loss and veneous congestion. This study aimed to determine the spatial relationship between the lesser saphenous vein and the cutaneous nerves, the venous anatomy in the lower leg, and the nerve distribution in the lateral dorsum of the foot, and we presented our clinical experience. MATERIALS AND METHODS: Twenty freshly amputated lower limbs were dissected in the 2 h following amputation. The lesser saphenous vein, medial/lateral sural nerve, and sural nerve were identified. Based on the anatomical studies, an island flap supplied by the vascular axis of the lesser saphenous vein and the lateral sural nerve was designed for clinical reparative applications in 24 cases. RESULTS: We indicated the spatial relationship between the lesser saphenous vein and the cutaneous nerves and the venous anatomy in the lower leg. Among 24 flaps, 21 showed complete survival (87.5%), while marginal flap necrosis occurred in two patients (8.33%) and distal wound dehiscence in another (4.17%). No symptomatic neuromas were observed. Their appearance and functioning were satisfactory, with filling maintained in the heel and lateral side of the foot. CONCLUSION: The distally based lesser saphenous veno-lateral sural neurocutaneous flap provides effective coverage of variable-sized soft tissue defects on the lower third of the lower leg and foot, without sensory loss and venous congestion.

[Imaging study on supratrochlear artery and its application in nasal reconstruction].

OBJECTIVE: To study the image of the muscular and cutaneous branches of supratrochlear artery by internal and external carotid angiography and to design the frontal flap for nasal reconstruction. METHODS: The muscular and cutaneous branches of supratrochlear artery of 30 adults were investigated through internal and external carotid angiography and three-dimensional reconstruction of vessels to explore the courses of branches of supratrochlear artery and their communication with supraorbital artery. According to image observation, the frontal flap was designed and used to perform nasal reconstruction in 11 cases of nasal defects, including 7 males and 4 females with an average age of 23 years (range, 15-48 years). The nasal defect ranged from 3.0 cm x 2.5 cm to 5.0 cm x 3.5 cm. RESULTS: Internal carotid angiography and three-dimensional imaging of the vessels showed that supratrochlear artery was found in 30 patients, with a diameter of (0.9 +/- 0.6) mm. The superficial cutaneous branch appeared constantly with a diameter of (0.7 +/- 0.3) mm. Deep muscular branch traveled among frontal muscle, with a diameter of (0.5 +/- 0.5) mm and a length of (32.0 +/- 6.2) mm, and it was missing in 4 patients, the branches communicated with each other and with supraorbital artery. All flaps survived with good appearance and without cartilage exposure. After 4 months to 3 years of follow-up, the postoperative aesthetic appearance and function of nasal tip, alar, and columella were satisfactory; the height and slope length of the external nose were moderate; and the reconstructed nose had good texture and normal ventilation function without constriction of naris. CONCLUSION: Imaging studies support the fact that supratrochlear artery has constant cutaneous branch. The application of frontal flap pedicled with cutaneous branch can improve the effectiveness of nasal reconstruction.

[The effect of adipose-derived stem cells on viability of random pattern skin flap in rabbits].

OBJECTIVE: To explore the effect of the adipose derived stem cells (ADSCs) on the survival of random pattern skin flap in rabbits. METHODS: ADSCs were isolated from fresh human fat and expanded in vitro for five passages. Then the characterization of ADSCs were determined by their CD marker profile and their ability to differentiate into osteogenic, adipogenic and chondrogenic lineages. On the back of the rabbits, two symmetric cephalic-based random pattern skin flaps were designed (6 cm x 2 cm). The right flaps were used as experimental groups with the contra-side flaps as control group. Human ADSCs were pre-labeled before seeding with fluorescent 3, 30-dioctadecyloxacarbocyanine perchlorate (DiO) dye. The experimental flaps evenly received 5-point injection of 2 x 10(6)Dio-labeled ADSCs resuspended in 0.5 ml of serum-free DMEM, while only 0.5 ml medium was injected into the control flaps. Seven days later, the survival rate of flaps was evaluated. The flaps underwent frozen section and were observed under the laser scanning confocal microscope to detect the fluorescence imaging. Flaps also underwent HE staining and were observed under light microscope to detect the vascular density. RESULTS: Compared with control group, there was a significant increase of flap survival rate in the experimental group (P < 0.01). Histological analysis also demonstrated a statistically significant increase in capillary density in the experimental group. CONCLUSIONS: It suggests that ADSCs have a better immune compatibility and potential for enhancing the blood supply of random pattern skin flaps.

[Evaluation of the role of serum vascular endothelial growth factor and urinary basic fibroblast growth factor in differential diagnosis of vascular anomalies].

OBJECTIVE: To evaluate the role of serum vascular endothelial growth factor (VEGF) and urine basic fibroblast growth factor (bFGF) in the differential diagnosis of vascular anomalies by receiver operating characteristic curve. METHODS: Using the method of enzyme linked immunosorbent assay (ELISA), 364 cases of various vascular anomalies (proliferating hemangiomas, n = 146; hemangiomas, n = 106; vascular malformations, n = 112) and 440 cases of various vascular anomalies (proliferating hemangiomas, n = 154; involuting hemangiomas, n = 148; vascular malformations, n = 138) subjects were examined for the serum levels of VEGF and the urine bFGF respectively. Nonparametric statistical tests were used for data analysis. The VEGF levels of proliferating hemangioma, involuting hemangioma, vascular malformation and control groups were analyzed by the Kruskal-Wallis test. When there was significance (P < 0.05), a subgroup analysis was performed by the Mann-Whitney U test. A receiver operator characteristic (ROC) curve was constructed for both the serum levels of VEGF and urine bFGF to determine the optimal diagnostic cut-off to differentiate proliferating hemangioma from involuting hemangioma, vascular malformation and controls. RESULTS: The serum level of VEGF and the urine bFGF in proliferating hemangiomas were significantly higher than those in involuting hemangiomas, vascular malformations and negative controls (P < 0.01). And the differences among the latter three groups were not statistically significant (P > 0.05). The optimal diagnostic cut-off point of serum VEGF and urine bFGF to differentiate proliferating hemangioma from involuting hemangioma, vascular malformation and controls was 99.6 pg/mg and 0.16 ng/mmol respectively. The area under ROC curve, the sensitivity and specificity were 99.7%, 99.3%, 99.7% and 91.9%, 98.7%, 71.1% respectively. The differences of the area under ROC curve of serum VEGF and urine bFGF showed no statistical significance (P > 0.05). CONCLUSION: Serum VEGF and urine bFGF are helpful for differentiating proliferating hemangioma from involuting hemangioma, vascular malformation and controls. Both parameters have higher values of clinical application.

[Experience of vaginal reconstruction by using a pudendal-thigh island flaps].

OBJECTIVE: To evaluate a pudendal-thigh island flap for vaginal reconstruction. METHODS: Forty-seven patients with congenital absence of vagina were undergoing the treatment. Based on the pedicle including the posterior labial neurovascular bundle, a pudendal-thigh island flap was designed and raised in the groin crease just lateral to the labia majora under the deep fascia. It was then transferred to the tunnel between the urethra and the anus for reconstruction of the vagina. RESULTS: From May of 1993 to July of 2001, 47 patients were successfully treated for the vaginal reconstruction with the flap. The results were satisfactory without complications. CONCLUSION: The above mentioned technique could be a safe and effect method for vaginal reconstruction with the advantages of reliable blood supply, good sensation and few complications. The areaes with the bilateral pudendal-thigh could be large enongh for the vaginal reconstruction without problem of the donor closure.

[Repair of the facial tissue defects with reversed narrow pedicle lateral maxillocervical fasciocutaneous flap].

OBJECTIVE: To introduce a method of repairing facial tissue defects caused by various factors. METHODS: The reverse narrow pedicle lateral maxillocervical fasciocutaneous flap was designed with its pedicle located in front of auricle. The size of the flaps ranged from 5 cm x 6 cm to 10 cm x 11 cm. The length and width of the pedicle ranged 2 cm-5 cm and 1 cm-1.5 cm. RESULTS: This fasciocutaneous flap was used in 21 cases. All flaps survived except 1 flap showing distal skin necrosis. CONCLUSION: This fasciocutaneous flap didn't include any major blood vessel. The ratio of the pedicle width to flap length broke that of traditional flap and was reverse transfer. The transposition of the narrow pedicle flap was easy. The postoperative appearance (color, texture, cosmetic aspect) was satisfactory. This method was a new design and was ideal for large facial tissue defect repair.