Co-Culture of Adipose-Derived Stem Cells and Chondrocytes With Transforming Growth Factor-Beta 3 Promotes Chondrogenic Differentiation.

Tissue engineering cartilage is a promising strategy to reconstruct the craniofacial cartilaginous defects. It demands plenty of chondrocytes to generate human-sized craniofacial frameworks. Partly replacement of chondrocytes by adipose-derived stem cells (ADSCs) can be an alternative strategy.The study aimed at evaluating the chondrogenic outcome of ADSCs and chondrocytes in direct co-culture with transforming growth factor-beta (TGF-ß3). Porcine ADSCs and chondrocytes were obtained from abdominal wall and external ears. Four groups: ADSCs or chondrocytes monocultured in medium added with TGF-ß3; ADSCs and ACs co-cultured with or without TGF-ß3. Cell growth rate was performed to evaluate the cell proliferation. Morphological, histologic and real-time polymerase chain reaction analysis were performed to characterize the chondrogenic outcome of pellets. ADSCs had favorable multi-lineage differentiation potential. Further, when ADSCs were co-cultured with chondrocytes in medium added with TGF-ß3, the cell proliferation was promoted and the chondrogenic differentiation of ADSCs was enhanced. We demonstrate that pellet co-culture of ADSCs and chondrocyte with TGF-ß3 could construct high quantity cartilages. It suggests that this strategy might be useful in future cartilage repair.

MicroRNA­214 promotes the EMT process in melanoma by downregulating CADM1 expression.

Melanoma is a malignant skin cancer type associated with a high mortality rate, but its treatment is currently not ideal. Both microRNA (miR)­214 and cell adhesion molecule 1 (CADM1) are differentially expressed in melanoma, but their role in this cancer type remains unknown. Therefore, the aim of the present study was to investigate the role of CADM1 and miR­214 in melanoma to identify novel targets for its treatment. The expression levels of CADM1 and miR­214 in cells were detected by reverse transcription­quantitative PCR (RT­qPCR). Moreover, cell viability, migration and invasion were measured by MTT, wound healing and Transwell assays, respectively. In addition, the relative expression levels of epithelial­mesenchymal transition (EMT)­related proteins in cells were detected by RT­qPCR and western blotting. It was found that the expression of CADM1 was inhibited in melanoma cells, while miR­214 expression was increased during melanoma tumorigenesis. Furthermore, miR­214 mimics promoted the viability, migration and invasion of melanoma cells. It was also demonstrated that the downregulation of CADM1 reversed the inhibitory effect of the miR­214 inhibitor in melanoma. Moreover, overexpression of CADM1 inhibited the EMT process in melanoma, while the miR­214 inhibitor suppressed the EMT process. The results also indicated that miR­214 promoted the EMT process by downregulating CADM1, which may represent a novel mechanism for the progression of melanoma.

Successful Rescue of the Victim Exposed to a Super High Dose of Iridium-192 during the Nanjing Radiological Accident in 2014.

Here we report on the interventions taken to treat a patient exposed to high-dose radiation and provide a protocol for treating such patients in the future. The patient, Mr. Wang, was a 58-year-old male janitor who was accidentally exposed to a 192Ir source with an activity of 966.4 GBq or 26.1 Ci. The dose estimated to the lower right limb was 4,100 Gy, whereas the whole-body effective dose was 1.51 Gy. The diagnosis was made according to the results of the patient dose estimation and clinical manifestations. Systemic treatment included stimulating bone marrow hematopoietic cells, enhancing immunity, anti-infection and vitamin supplements. The treatment of radiation-induced skin lesions consisted of several debridements, two skin-flap transplantations and application of mesenchymal stem cells (MSCs). Skin-flap transplantations and MSCs play important roles in the recovery of skin wound. A combination of antibiotics and antimycotic was useful in reducing inflammation. The application of vacuum sealing drainage was effective in removing necrotic tissue and bacteria, ameliorating ischemia and hypoxia of wound tissue, providing a fresh wound bed for wound healing and improving skin or flap graft survival rates. The victim survived the accident without amputation, and function of his highly exposed right leg was partially recovered. These results demonstrate the importance of collaboration among members of a multidisciplinary team in the treatment of this patient.

Fluid management in extensive liposuction: A retrospective review of 83 consecutive patients.

Tumescent anesthesia makes it feasible to perform liposuction in an office setting. There are often patients who desire extensive liposuction on approximately 30% of total body surface area, which means the potential of fluid overload. In this study, the charts of 83 patients undergoing extensive liposuction were retrospectively reviewed. The intra-operative fluid ratio was 1.66 for the extensive liposuction. There were no episodes of pulmonary edema, congestive heart failure exacerbation, or other major complications. The average urine output in the operating room, the recovery room, and while on the floors was 1.35, 2.3, and 1.4 mL/kg/hour respectively. Intravenous (IV) fluid administration during operation was minimized to approximately 300 to 500 mL. The total volume of IV injection was also reduced to less than 1500 mL when the patient was in the recovery room and on the hospital floor. Our fluid management strategy in extensive liposuction reflects minimal risk of volume overload. Foley catheters are not applied and patients could resume oral intake in usual, so they can discharge after 6 hours of recovery room stay in our daily practice.

GDC-0084 inhibits cutaneous squamous cell carcinoma cell growth.

GDC-0084 is a novel and potent small-molecule PI3K-mTOR dual inhibitor. The present study examined its potential activity in cutaneous squamous cell carcinoma (cSCC) cells. Our results show that GDC-0084 treatment at nanomole concentrations potently inhibited survival and proliferation of established (A431, SCC-13 and SCL-1 lines) and primary human cSCC cells. GDC-0084 induced apoptosis activation and cell cycle arrest in the cSCC cells. It was more efficient than other known PI3K-Akt-mTOR inhibitors in killing cSCC cells, but was non-cytotoxic to the normal human skin fibroblasts/keratinocytes. In A431 cells and primary cSCC cells, GDC-0084 blocked phosphorylation of key PI3K-Akt-mTOR components, including p85, Akt, S6K1 and S6. GDC-0084 also inhibited DNA-PKcs activation in cSCC cells. Significantly, restoring DNA-PKcs activation by a constitutively active-DNA-PKcs (S2056D) partially inhibited GDC-0084-induced cell death and apoptosis in A431 cells. In vivo, GDC-0084 daily gavage potently inhibited A431 xenograft tumor growth in mice. In GDC-0084-treated tumor tissues PI3K-Akt-mTOR and DNA-PKcs activation were significantly inhibited. In summary, GDC-0084 inhibits human cSCC cell growth in vitro and in vivo through blocking PI3K-Akt-mTOR and DNA-PKcs signalings.

Bromodomain protein BRD4 promotes cell proliferation in skin squamous cell carcinoma.

The present study examined the expression and biological functions of bromodomain-containing protein 4 (BRD4) in skin squamous cell carcinoma (SCC) cells. Our results show that BRD4 mRNA and protein expression was upregulated in human skin SCC cells, as compared to its level in the normal skin keratinocytes and fibroblasts. Treatment with BRD4 inhibitors, JQ1 and CPI203, resulted in proliferation inhibition, apoptosis and cell cycle arrest in both established (A431 cell line) and primary skin SCC cells. Furthermore, BRD4 knockdown (by targeted shRNAs) or knockout (by CRISPR/Cas9) largely inhibited A431 cell proliferation. Reversely, forced-overexpression of BRD4 in A431 cells facilitated cell proliferation. We show that BRD4 is required for the expression of several oncogenes, including cyclin D1, Bcl-2 and MYC. BRD4 inhibition, knockdown or knockout significantly decreased above oncogene expression in SCC cells. In vivo, CRISPR/Cas9-mediated BRD4 knockout significantly suppressed A431 xenograft tumor growth in severe combined immunodeficient (SCID) mice. Together, our results suggest that BRD4 could be a novel and pivotal oncogenic protein of skin SCC.

A method for reconstruction of vermilion tubercle.

BACKGROUND: This article reports on the bilateral lip mucosa flaps (BLMFs) for reconstruction of the vermilion tubercle of the corrected cleft lip deformity. METHODS: From 2005 to 2013, the bilateral mucosa flaps procedures were performed on 34 secondary cleft lip patients (bilateral, n = 25; unilateral, n = 9). We retrospectively reviewed the pre-, intra-, and postoperative data of these 34 cases receiving BLMFs. RESULTS: Satisfactory results were obtained, both cosmetically and functionally, in all 34 patients. Lateral projection measurements of the reconstructive vermilion tubercle showed a mean increase of 151%. There were minimal perioperative complications. The color and texture matched well with the surrounding tissue. The postoperative scars are not remarkable at all. CONCLUSIONS: The technique is relatively simple. The BLMFs provided a versatile and reliable option for the reconstruction of the vermilion tubercle from secondary cleft lip deformities.

[Experimental study of the effect of deferasirox on the micro-angiogenesis in narrow pedicle flap through epithelial-mesenchymal transition].

OBJECTIVE: To investigate the effect of Deferasirox on the micro-angiogenesis in narrow pedicle flap through Epithelial-Mesenchymal Transition. METHODS: 32 male rats were randomly divided into group I and II which were subdivided into Ia and Ib, IIa and IIb, 8 rats in each group. The rats were administrated intragastrically for 7 days with Deferasirox 100 mg/kg in group Ia and IIa, with the same dose of N. S. in group Ib and IIb. After that, narrow pedicle flaps were formed on the rats back. In group I, the subcutaneous vascular network was observed intraoperatively. The flap survival rate was recorded. In group II , specimens were collected at the distal end of flaps 3 days after operation. IHC and Western Blot were done to examine the expression of CD34, E-cadherin, Vimentin. The microvessel density was also calculated. RESULTS: The subcutaneous micro-angiogenesis in group Ia was more exuberant than that in group Ib. The narrow pedicle flaps in group Ia survived completely, while the survival rate was 62.5% in group Ib (P < 0.05). The percentage of flap survival area for Ia and Ib was (100 +/- 0.00) % and (84.06 +/- 4.42)% (P < 0.05). The expression of E-cadherin in IIa was lower than that in IIb, while the expression of Vimentin and CD34 were higher in IIa, showing statistically difference (P < 0.05). CONCLUSION: Deferasirox can improve the flap micro-angiogenesis through inducing epithelial-mesenchymal transition, so as to improve the survival rate of narrow pedicle flap.

[Application of slender narrow pedicle flap in facial skin cancer treatment].

OBJECTIVE: To introduce the application of slender narrow pedicle flap in repairing facial tissue defects after skin carcinoma excision, and investigate its survival mechanism. METHODS: The slender narrow pedicle iateral maxillocevical flap was designed with its pedicle including skin fascia or only the fascia located in front of auricle or behind of it, repaired 26 cases of facial defects, including 5 temporal skin basal cell carcinoma, 6 skin squamous cell carcinoma, and 1 facial skin malignant melanoma, 8 skin basal cell carcinoma, 5 skin squamous cell carcinoma, 1 skin mucinous carcinoma. In 26 cases, 24 cases their pedicles in front of the auricle, 2 cases behind of the auricle; 4 cases their pedicles only including fascia. The size of the flaps ranged from 3.0 cm x 2.5 cm to 10.0 cm x 8.0 cm. The width and length of the pedicle ranged 1.0-1.5 cm and 2-6 cm. RESULTS: 26 cases of the slender narrow pedicle flaps all survived and the results were satisfactory except 5 cases of distal congestion, then gradual recovery. CONCLUSIONS: This slender narrow flap don't include any major blood vessel, without dissecting the blood vessels in operating. Due to its slender pedicle, the whole flap looks like "pingpang bat", the flap rotation is easy and its coverage area is very large, without cat ears. The postoperative appearance (color, texture, cosmetic aspect) is satisfactory. This slender narrow flap is an extraordinary new flap design and is ideal for the repair of the facial tissue defect after skin carcinoma excision.

[Experiment study of SDF-1 expression during the survival process of the narrow pedicle flap with hypoxia and ischemia].

OBJECTIVE: To detect the stromal cell derived factor 1 (SDF-1) expression during the survival process of the narrow pedicle flap with hypoxia and ischemia and to investigate the role of SDF-1/ CXCR4 axis in flap neovascularization. METHODS: The narrow pedicle flaps were formed on the bilateral back of 5 pigs. The pedicle ratio of length to width was 4:2. The flap size was 2 cm x 2 cm (group A), 3 cm x 3 cm (group B), 4 cm x 4 cm (group C), 5 cm x 5 cm(group D), 6 cm x6 cm (group E). The flaps survival rate was observed and HE staining was performed. The SDF-1 expression at the distal end of flaps was detected by ELISA during the operation and 3, 5, 7, 14 days after operation. RESULTS: (1) SDF-1 expression at the same group increased after operation until it reached the peak value at 5 days after operation; then it decreased to basic value. (2) SDF-1 expression in different groups was higher in bigger flaps until the flaps size reached 5 cm x 5 cm. Then partial necrosis happened at the distal end of flaps. CONCLUSIONS: The SDF-1 expression may be related to the blood supply during the survival process of the narrow pedicle flap with hypoxia and ischemia.

The application of bilateral lip mucosa flaps in repairing upper vermilion defects.

BACKGROUND: The lip is a primary aesthetic feature of the human face. Repair of vermilion defects represents a unique challenge to the reconstructive surgeon. The methods of repair are various and have both advantages and disadvantages. Bilateral lip mucosa flaps were applied to the repair of upper vermilion defects, and the effects were observed and reported in this article. METHODS: The bilateral lip mucosa flaps were designed. Two reverse triangular flaps were designed on both sides of the upper vermilion defect. These 2 flaps have the common pedicle in the vermilion depression site. Alternatively, a "λ" incision may be used in the wet vermilion. Then, the mucosa flaps were raised, and the dissection was performed between the orbicularis oris muscles and oral mucosa glands. Next, the bilateral mucosa flaps were rotated down 90° and inserted into the "λ" incision to increase the tissue volume of the middle upper lip and deepen the labiogingival sulcus. The common pedicle of the mucosa flaps was formed to the vermilion tubercle, and the incisions were sutured layer by layer. RESULTS: From 2005 to 2010, the bilateral mucosa flaps procedures were performed on 30 secondary cleft lip patients (bilateral, n=22; unilateral, n=8). Lateral projection measurements of the reconstructive vermilion tubercle showed a mean increase of 149%. Satisfactory results were obtained, both cosmetically and functionally, in all 30 patients. There were minimal perioperative complications. The postoperative scars are not remarkable at all. CONCLUSIONS: The technique is relatively simple. The bilateral lip mucosa flaps provided a versatile and reliable option for the correction of vermilion defects from secondary cleft lip deformities.

[Experimental study on the relationship between the ratio of length to width of slender narrow pedicle and random flap survival area in pig].

OBJECTIVE: To explore the relationship between the ratio of length to width of slender narrow pedicle and random flap survival area, and to provide a theoretical basis for the clinical application of slender narrow pedicle flaps. METHODS: 25 pigs were randomly divided into 5 groups, 5 pigs in each group. The ratio of length to width of slender narrow pedicle in 5 groups respectively was: 0:2, 1:2, 2:2, 3:2, 4:2. Every ratio' s slender narrow pedicle was carrying five different size of random flaps, which were 2 cm x 2 cm(A), 3 cm x 3 cm(B),4 cm x 4 cm(C), 5 cm x 5 cm(D), 6 cm x 6 cm(E), respectively. Flap A was control flap. In each group, flap A, B, C, D and E were created in each pigs' bilateral back. The order in both sides back is contrary. The flaps were evaluated with the general observation, fluorescence examination, blood flow ECT test, pathological expression and computerized analysis of survival area. RESULTS: (1) The living process and pathologic process of traditional flap and slender narrow pedicle flap were consistent. It could not postpone the flap living process when the flap pedicle became long and narrow. (2) When the ratio of the length to width of the slender narrow pedicle was constant, along with the flap area increased, the flap survival area also increased, but when the flap reached a certain area , the distal flap would necrosis,the flap survival area would not reduce. (3) When the flap size remained unchanged, along with the ratio of the length to width of the slender narrow pedicle increased, the flap survival area was not affected, but when the ratio of the length to width of the slender narrow pedicle increased to a certain limit, distal flap would necrosis, the flap survival area would reduce. CONCLUSIONS: (1) Pedicle width of random flap can be much smaller than flap width. The ratio of pedicle width to flap length is far less than traditional ratio. (2) The pedicle of random flap can be designed as slender shape, so that the whole flap looks like "pingpang bat", which makes the narrow pedicle flap rotate easily. (3) A certain ratio of the length to width of a slender narrow pedicle has a maximum flap survival area, and increasing the flap size or ratio of the length to width of a slender narrow pedicle in a certain extent will not lead to flap necrosis.

[Island sternocleidomastoid myocutaneous flap for repairing the buccal composite tissue defect].

OBJECTIVE: To study the application of island sternocleidomastoid myocutaneous flap in repairing the buccal composite tissue defect especially penetrated tissue defect. METHODS: The flap pedicle included upper part OF sternocleidomastoid muscle and occipital artery. The rotation point was located at 2 cm below the mastoid. The distance between the pivot point and distal border of the defect was the length of the muscular flap. The width of the flap was slightly larger than the defect, but should not be more than 7 cm. The lower border of the flap should not exceed 2 cm below the collar bone. The flap was elevated from the starting point of the sternocleidomastoid muscle and beneath it. The pedicle only contained muscle. The flap was transferred to the defect through the tunnel between the pedicle and defect. The wounds at donor sites were closed directly or with skin graft or local flaps. RESULTS: 12 cases were treated, including 10 cases of buccal soft tissue tumors and 2 cases of buccal penetrated defects. All the musculocutaneous flaps survived with good texture, color and thickness. CONCLUSIONS: The island sternocleidomastoid myocutaneous flap is an ideal flap for large buccal composite tissue defect with reliable blood supply. It is easily performed and very practical.

[Clinic application of compound flap pedicled with arterial arch of palpebral margin].

OBJECTIVE: To study the clinic application of compound flap pedicled with arterial arch of palpebral margin in repairing severe full defect of eyelid. METHODS: According to eyelid structure and the defect size, the two compound flaps were designed beside the defect based on the arterial arch of the palpebral margin. If the defective area was too large, the lateral compound flap may be extended to lower or upper eyelid 0.5 cm away from the outer canthus, then cut and propelled the two compound flaps to repair the full eyelid defect. RESULTS: 20 cases had been cured with this method since 1998. In this cases, 4 cases were basal cell carcinoma of eyelid, 2 cases were squamous carcinoma, 3 angiomas, 6 chromatophore nexuses, 3 traumatic defects, 2 congenital defects. The largest length of eyelid full defect was 1.7 cm and the smallest was 0.8 cm. 6 cases were upper eyelid defect and 14 cases were lower eyelid defect. All the compound flaps survived completely without any complications. All cases obtained satisfactory results functionally and esthetically. CONCLUSIONS: Repairing full eyelid defect with the compound eyelid flap is the same kind tissue repairing. It can not only provide enough tissues to primary repair large full defect of the upper or lower eyelid to restore normal anatomical structure and appearance of the eyelid, but also is easy to be operated without severe secondary deformities. The arterial arch of the palpebral margin is constant and the blood supply of the compound flap is reliable. It is an ideal method of repairing the eyelid defect.

[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.