M2-polarized macrophages mediate wound healing by regulating connective tissue growth factor via AKT, ERK1/2, and STAT3 signaling pathways.

BACKGROUND: Timely and sufficient M1 recruitment and M2 polarization are necessary for fibrosis during wound healing. The mechanism of how M2 mediates wound healing is worth exploring. Abnormally up-regulated connective tissue growth factor (CTGF) influences multiple organ fibrosis, including cardiac, pulmonary, hepatic, renal, and cutaneous fibrosis. Previous studies reported that M2 contributed to hepatic and renal fibrosis by secreting CTGF. It is worth discussing if M2 regulates fibrosis through secreting CTGF in wound healing. METHODS AND RESULTS: We established the murine wound model and inhibited macrophages during proliferation phase with clodronate liposomes in vivo. Macrophages depletion led to down-regulation of wound healing rates, collagen deposition, as well as expression of collagen 1/3 and Ki67. M2 was induced by interleukin-4 (IL-4) and measured by flow cytometry in vitro. Secreted pro-fibrotic and anti-fibrotic factors were tested by enzyme-linked immunosorbent assay (ELISA). M2 was polarized, which producing more CTGF, transforming growth factor-beta1 (TGF-ß1), and IL-6, as well as less tumor necrosis factor-α (TNF-α) and IL-10. M2 CTGF gene was blocked using siCTGF. Effects of M2 on fibroblasts activities were detected by cell counting kit 8 (CCK8) and cellular wound healing assay. Expressions of related signaling pathway were assessed by western blotting. Blockade of CTGF in M2 deactivated fibroblasts proliferation and migration by regulating AKT, ERK1/2, and STAT3 pathway. Recombinant CTGF restored these effects. CONCLUSIONS: Our research, for the first time, indicated that M2 promoted wound healing by secreting CTGF, which further mediating proliferation and migration of fibroblasts via AKT, ERK1/2, and STAT3 pathway.

Knockdown of DEAD-box RNA helicase 52 (DDX52) suppresses the proliferation of melanoma cells in vitro and of nude mouse xenografts by targeting c-Myc.

The ATP-dependent protein DEAD-box RNA helicase 52 (DDX52) is an important regulator in RNA biology and has been implicated in the development of prostate and lung cancer. However, its biological functions and clinical importance in malignant melanoma (MM) are still unclear. Understanding the potential mechanism underlying the regulation of MM progression by DDX52 might lead to novel therapeutic strategies. The aim of the present study was to investigate the role of DDX52 in the regulation of MM progression and its clinical relevance. DDX52 expression in normal and MM tissues was evaluated by GEO analysis and immunohistochemistry. The effects of DDX52 on cell growth were evaluated in MM cells with downregulated DDX52 expression. In this study, we found that DDX52 was markedly overexpressed in MM tissues compared with nontumor tissues and was associated with shorter overall survival in patients; therefore, DDX52 might be a prognostic marker in MM. Downregulation of DDX52 expression in the MM cell lines A2058 and MV3 markedly inhibited cell proliferation and colony formation. Additionally, knockdown of DDX52 in MM cells caused significant regression of established tumors in nude mice and delayed the onset time. Moreover, downregulation of DDX52 markedly suppressed c-Myc mRNA and protein expression, and an RNA immunoprecipitation assay confirmed the association between DDX52 and c-Myc. Restoration of c-Myc expression partly rescued the effects of DDX52 deficiency in MM cells. In conclusion, our study found that DDX52 mediated oncogenesis by promoting the transcriptional activity of c-Myc and could be a therapeutic target in MM.

Circular RNA circ_0020710 drives tumor progression and immune evasion by regulating the miR-370-3p/CXCL12 axis in melanoma.

BACKGROUND: Circular RNAs (circRNAs) have been reported to have critical regulatory roles in tumor biology. However, their contribution to melanoma remains largely unknown. METHODS: CircRNAs derived from oncogene CD151 were detected and verified by analyzing a large number of melanoma samples through quantitative real-time polymerase chain reaction (qRT-PCR). Melanoma cells were stably transfected with lentiviruses using circ_0020710 interference or overexpression plasmid, and then CCK-8, colony formation, wound healing, transwell invasion assays, and mouse xenograft models were employed to assess the potential role of circ_0020710. RNA immunoprecipitation, luciferase reporter assay and fluorescence in situ hybridization were used to evaluate the underlying mechanism of circ_0020710. RESULTS: Our findings indicated that circ_0020710 was generally overexpressed in melanoma tissues, and high level of circ_0020710 was positively correlated with malignant phenotype and poor prognosis of melanoma patients. Elevated circ_0020710 promoted melanoma cell proliferation, migration and invasion in vitro as well as tumor growth in vivo. Mechanistically, we found that high level of circ_0020710 could upregulate the CXCL12 expression via sponging miR-370-3p. CXCL12 downregulation could reverse the malignant behavior of melanoma cells conferred by circ_0020710 over expression. Moreover, we also found that elevated circ_0020710 was correlated with cytotoxic lymphocyte exhaustion, and a combination of AMD3100 (the CXCL12/CXCR4 axis inhibitor) and anti-PD-1 significantly attenuated tumor growth. CONCLUSIONS: Elevated circ_0020710 drives tumor progression via the miR-370-3p/CXCL12 axis, and circ_0020710 is a potential target for melanoma treatment.

Hsa-let-7b Suppresses Cell Proliferation by Targeting UHRF1 in Melanoma.

UHRF1 promotes melanoma progression by inducing cell proliferation, and is correlated with poor prognosis of melanoma patients. However, the regulation mechanism has not been fully elaborated. Here, we detected hsa-let-7b expression and its role in melanoma. Through Targetscan and miRanda predication, 30 overlapped miRNAs were found; further survival analysis revealed that hsa-let-7b was the only miRNA that affected the overall survival. Overexpressed hsa-let-7b could significantly inhibit the proliferation ability of A375 and A2058 cells, and this phenomenon was reversed after co-transfection with pLenti-UHRF1. In conclusion, hsa-let-7b regulates melanoma cells proliferation in vitro by targeting UHRF1.

Elevated kindlin-2 promotes tumour progression and angiogenesis through the mTOR/VEGFA pathway in melanoma.

BACKGROUND: In our previous study, kindlin-2 promoted skin wound healing and decreased the permeability of neovascularization during angiogenesis. Herein, we explored the biological function and underlying mechanism of kindlin-2 in cutaneous melanoma. METHODS AND RESULTS: Through a series of in vitro assays, we found that high levels of kindlin-2 promoted migration and invasion of melanoma cells without influencing cell proliferation. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analyses showed that upregulated kindlin-2 promoted the cellular epithelial-mesenchymal transition (EMT). Importantly, we found that melanoma cells overexpressing kindlin-2 promoted angiogenesis and VEGFA secretion in vitro and facilitated tumour growth and lung metastasis in vivo. To unveil the underlying mechanism, we conducted Next-generation sequencing (NGS) and differential expression analyses. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that overlapping differentially expressed genes (DEGs) were primarily enriched in the TGF-ß, mTOR and VEGF signalling pathways. Then, we confirmed that the mTOR/VEGFA pathway was activated during the process of kindlin-2-induced melanoma progression and angiogenesis. Moreover, we demonstrated that kindlin-2 was significantly overexpressed in clinical melanoma samples and that a high level of kindlin-2 predicted a poor prognosis. CONCLUSIONS: Taken together, these findings showed that kindlin-2 promotes angiogenesis and tumour progression via the mTOR/VEGFA pathway.

TRIM44 activates the AKT/mTOR signal pathway to induce melanoma progression by stabilizing TLR4.

BACKGROUND: There is growing evidence that tripartite motif-containing protein 44 (TRIM44) plays crucial role in tumor development. However, the underlying mechanism of this deubiquitinating enzyme remains unclear. METHODS: Large clinical samples were used to detect TRIM44 expression and its associations with clinicopathological features and prognosis. Gain- and loss-of-function experiments in cell lines and mouse xenograft models were performed to elucidate the function and underlying mechanisms of TRIM44 induced tumor progression. Co-immunoprecipitation (Co-IP) assays and mass spectrometric analyses were applied to verify the interacting proteins of TRIM44. RESULTS: We found that TRIM44 was commonly amplified in melanoma tissues compared with paratumoral tissues. TRIM44 expression also positively correlated with more aggressive clinicopathological features, such as Breslow depth (p = 0.025), distant metastasis (p = 0.012), and TNM stage (p = 0.002). Importantly, we found that TRIM44 was an independent indicator of prognosis for melanoma patients. Functionally, overexpression of TRIM44 facilitated cell invasion, migration, apoptosis resistance and proliferation in vitro, and promoted lung metastasis and tumorigenic ability in vivo. Importantly, high level of TRIM44 induced melanoma cell epithelial-mesenchymal transition (EMT), which is one of the most important mechanisms for the promotion of tumor metastasis. Mechanistically, high levels of TRIM44 increased the levels of p-AKT (T308) and p-mTOR (S2448), and a specific AKT inhibitor inhibited TRIM44-induced tumor progression. Co-IP assays and mass spectrometric analyses indicated that TRIM44 overexpression induces cell EMT through activating AKT/mTOR pathway via directly binding and stabilizing TOLL-like receptor 4 (TLR4), and TLR4 interference impeded TRIM44 induced tumor progression. Moreover, we demonstrated that TRIM44 is the target of miR-26b-5p, which is significantly downregulated in melanoma tissues and may be responsible for the overexpression of TRIM44. CONCLUSIONS: TRIM44, regulated by miR-26b-5p, promotes melanoma progression by stabilizing TLR4, which then activates the AKT/mTOR pathway. TRIM44 shows promise as a prognostic predictor and a therapeutic target for melanoma patients.

Downregulation of RNF128 activates Wnt/ß-catenin signaling to induce cellular EMT and stemness via CD44 and CTTN ubiquitination in melanoma.

BACKGROUND: Ring finger proteins (RNFs) were involved in carcinogenesis. Here, we aimed to explore the detailed mechanism of RNF128 in the progression of melanoma. METHODS: We reanalyzed several gene expression profiles from the Gene Expression Omnibus (GEO) database and obtained the overlapped differential expressed RNF genes. Among them, RNF128 was selected to further explore its expression, the biological significance, and the underlying molecular mechanism, as well as the clinical relevance in melanoma patients. RESULTS: RNF128 was found to be significantly downregulated in the selected datasets, which was further verified in our melanoma tissues. Moreover, RNF128 downregulation was shown to correlate with the malignant phenotype of melanoma, and further functional assays demonstrated that low levels of RNF128 promoted melanoma progression via inducing cell epithelial-mesenchymal transition (EMT) and the acquisition of stemness. Mechanistically, RNF128 interference activated the Wnt pathway via simultaneously ubiquitinating CD44/cortactin (CTTN), resulting in CD44 and c-Myc transcription, thus revealed that RNF128 participated in a positive feedback of the Wnt pathway-CD44 loop. Clinically, we found that patients expressing low RNF128 and high CD44/CTTN levels had a poor prognosis. CONCLUSION: Downregulated RNF128 activates Wnt signaling to induce cellular EMT and stemness by ubiquitinating and degrading CD44/CTTN, and RNF128 is a reliable diagnostic and prognostic biomarker, and a deeper understanding of RNF128 may contribute to the treatment of melanoma.

Hair follicle units promote re-epithelialization in chronic cutaneous wounds: A clinical case series study.

Chronic cutaneous wounds are one of the most unfavorable pathophysiological processes in routine practice. However, developments in hair follicle unit therapy may aid the treatment of these wounds. The aim of the present study was to investigate the function of hair follicle units in chronic cutaneous wound re-epithelialization and to develop an effective protocol for wound treatment. A total of 14 patients, of which nine were male and five were female, with a mean age of 60.71 years (range, 19-76 years) and a mean wound area of 74.14 cm2, were treated in the study. The hair follicle units were dissected from a scalp graft and transplanted into the chronic cutaneous wound bed, after which clinical evaluation was performed. Images of the recipient site were captured at 0, 1, 2, 3, 4, 5, 8 and 14 weeks following transplantation. In addition, histological examinations were conducted postoperatively at week 16. Total wound re-epithelialization was observed in all the patients. Histological analysis revealed that the epidermis and papillary dermis were present in the wound area. Adnexal structures and the reticular dermis were also observed. Therefore, the present study demonstrated the ability of hair follicle units to promote chronic cutaneous wound healing.

Bilateral reduction mammoplasty after heart transplantation.

We report a bilateral reduction mammoplasty in a 15 year old female who suffered increasing back and shoulder pain and chest wall discomfort associated with bilateral breast enlargement during a 17 month period following heart transplantation. Cardiologic evaluation confirmed a structurally normal heart with good systolic and diastolic function, and ejection fraction of 80%. We performed a bilateral mammoplasty using dermal suspension flap in vertical-scar reduction. The patient recovered satisfactorily without incident, and breast morphology was excellent at the 2 year 9 month follow-up, with no recurrence of her previous symptoms or further hyperplasia.

Treatment of sternal wound infections after open-heart surgery.

OBJECTIVE: The aim of this study was to investigate the proper treatment of infected median sternotomy wounds. METHODS: A retrospective study was conducted to investigate the proper treatment of infected median sternotomy wounds on patients with sternal wound infections from January 2007 to July 2009. The characteristics of the sternal infections and the treatment outcomes were analysed. RESULTS: Ninety-seven patients with sternal wound infections were treated. A total of 32 patients acquired the infection within one month after open-heart surgery, 10 patients got the infection one to two months after the surgery, and 1 patient died two days after debridement. There were 54 patients who acquired the infection beyond two months post-surgery, while 1 patient died on the day before the operation. One patient received four cycles of wound debridement, 18 patients received two cycles and 78 patients only received one cycle. A total of 14 patients received a vacuum-assisted closure treatment. There were 73 patients who had surgery for repair of muscle flaps, 1 patient for breast tissue flap, 63 patients for pectoralis major muscle flap, and 9 patients with rectus abdominis muscle flap. There were 12 patients who received a transverse plate fixation of the sternum with titanium plating. CONCLUSION: A positive prognosis can be obtained by the algorithm treatment based on the onset and depth of the sternal infection.

[The treatment of sternal wound infection by internal fixation of sternum with titanium plate after cardiac surgery].

OBJECTIVE: To investigate the internal fixation of sternum with titanium plate for the treatment of sternal wound infection after cardiac surgery. METHODS: A retrospective study of 26 patients from Jan 2009 to Dec 2011 was carried out. All the 26 cases with sternal wounds infection after cardiac surgery, were treated by internal fixation with titanium plates. RESULTS: All the patients had chronic infection, lasting for more than 3 months. 3 patients received vacuum-assisted closure treatment after complete debridement. All the wounds were covered with pectoral major muscle flap. All the wounds healed completely with no recurrence of infection. CONCLUSIONS: Internal fixation of sternum with titanium plate can increase the stability. It helps to improve the treatment efficacy of sternal wound infection after cardiac surgery.

Plastic surgery after solid organ transplantations.

BACKGROUND: More patients receive organ transplantation surgeries due to the advancement in immunosuppressive agents and surgical techniques. Some of those patients may need to undergo plastic or reconstructive surgery. Long-term use of immunosuppressive agents raises some serious problems. Therefore, this study aimed to introduce our experience about the safety and effectiveness of plastic surgeries after solid organ allograft transplantation. METHODS: A retrospective review of 17 transplant recipients who underwent different reconstructive or cosmetic operations was carried out. The subjects included 1 heart transplant, 1 liver transplant and 15 kidney transplant recipients. RESULTS: All patients tolerated the plastic surgery procedures well. Flaps and skin grafts were the main constructive methods. There were no postoperative infections and wound dehiscence. Transferred flaps survived completely. Skin grafts took well. Three of the cosmetic surgery patients were satisfied with the results. CONCLUSIONS: Immunosuppressed organ transplant recipients can successfully undergo major reconstructive and cosmetic surgery when given special attention.

[Vertical reduction mammaplasty with wide superior pedicle].

OBJECTIVE: To report the reduction mammaplasty with vertical incision and superior wide pedicle. METHODS: Typical Lejour mosque-dome design was performed. The inferior part of glandular tissue and skin were excised. The nipple-and-areola complex (NAC) was elevated to normal position with superior wide pedicle. The breast morphology was modified with vertical scar left. RESULTS: 46 patients were treated. 4 patients had unilateral breast reduction. 14 breasts had wound dehiscence. 3 breasts received debridement, others were treated conservatively with dressings. No complete NAC necrosis occurred. CONCLUSIONS: The reduction mammaplasty with vertical incision and superior wide pedicle is a safe and effective method with a low risk of NAC necrosis.

[A retrospective study of 129 cases with immediate breast reconstruction after skin-sparing mastectomy for breast cancer].

OBJECTIVE: To evaluate the oncologic safety, indications and aesthetic results for skin-sparing mastectomy (SSM) and immediate breast reconstruction (IBR). METHOD: One hundred and twenty-nine breast cancer patients treated by SSM + IBR from October 1999 to May 2007 were reviewed. Reconstructive techniques included latissimus dorsi flaps (38 patients), implants only (2 patients), latissimus dorsi flaps plus implants (61 patients), pedicled transverse rectus abdominis myocutaneous (TRAM) flaps (25 patients) and deep inferior epigastric artery perforator (DIEP) flaps (3 patients). Aesthetic results were judged by patients' self-evaluation. RESULTS: Mean duration of hospitalization was 18.6 days. Time of first chemotherapy was 5.2 days after operation. Eleven patients (11/63, 17.5%) developed capsular contracture and 24 patients (24/99, 24.2%) developed seroma in the donor site. Nine patients (9/28, 32.1%) developed partial fat necrosis in TRAM and DIEP flaps. The satisfaction with the aesthetic results of the reconstructive breast was significantly lower in irradiated patients than non-irradiated ones. Median follow-up time was 11 months. Five patients developed local recurrence and 7 patients with metastasis. CONCLUSIONS: SSM with IBR can be used for the 0 to II a stage breast cancer patients, with surgical oncologic and aesthetic satisfaction. Radiotherapy has an adverse effect on the reconstructive breast. Delayed or delayed-immediate reconstructions are recommended for patients indicated to postoperative radiotherapy.

[The study on donor site sequelae after autologous breast reconstruction with an extended latissimus dorsi flap].

OBJECTIVE: To study the prevention and treatment on the donor site sequelae after autologous breast reconstruction with extended latissimus dorsi flap. METHODS: 88 patients received breast reconstruction with extended latissimus dorsi flap between May 1999 and Nov 2004 were concerned. We analyzed the donor site sequelae by objective and subjective evaluation and we assessed the functional condition. Results Dorsal hematoma developed in 1 patient. There are 7 patients with dorsal seromas and 2 of them developed capsules; 5 patients with mild skin exfoliate and 1 patient with dry skin necrosis, 2 winged shoulders. No infection and no hypertrophic scars appeared in donor site. There is no significant functional limitation and no influence in daily life. CONCLUSIONS: The main complication after autologous breast reconstruction with extended latissimus dorsi flap was seromas and it can be controlled to decrease. There is no severe complication. It's a worthy method in autologous breast reconstruction in oriental people.

[Vertical reduction mammaplasty].

OBJECTIVE: The inverted "T" operation is a typical and classic method for breast hypertrophy. Although having good results, it leaves significant scar and tends to have bottom-out deformity with time. The purpose of this study is to introduce our experience in vertical mammaplasty. METHODS: The typical Lejour's design was used. The lower part of the breast was excised. The nipple-alveolar complex was elevated to a proper position with the superior dermal-glandular pedicle. Only vertical scar was left after the skin was closed. RESULTS: 24 patients were treated using this method. The results were satisfactory except unilateral nipple-alveolar complex necrosis occurred in one patient. Another patient had a minor revision for the dog-ear 6 months after breast reduction. CONCLUSIONS: Vertical reduction mammaplasty is effective with reduced scar and highly satisfactory results. It deserves recommendation.

[Evaluation of a compound with dan-shen root and azone for scar treatment].

OBJECTIVE: To evaluate the effect of the compound of Dan-shen root and azone for scar treatment. METHODS: The rat skin in vitro and the human skin in vitro and vivo were separately examined their permeability of the mixture of the Dan-shen root and azone. The 301 patients with hypertrophic scar were randomly divided into two groups: one treated with elastic cloth paste (including silicone) contained in Dan-shen root with azone, and the another treated with only elastic cloth paste (including silicone). RESULTS: The permeability of Dan-shen root with azone, passing through the rat skin in vitro and the human skin in vitro and vivo was significantly higher than both the distilled water and the normal saline (P < 0.05). In the clinical study for treatment of the hypertrophic scars, the efficient rate of the group with the Dan-shen root with azone was significantly higher than the control (91.4% vs. 71.3%) (P < 0.01). CONCLUSION: The Dan-shen root with azone could be easier to permeate the skin and more effective to treat the hypertrophic scar.

[Flap transplantation combined with liposuction to treat upper limb lymphedema after mastectomy].

OBJECTIVE: To investigate a new surgical method to treat unilateral limb lymphedem after radical mastectomy. METHODS: 10 cases of upper limb lymphedema after radical mastectomy were treated using flap transfer (the lateral thoracic skin flap or latissimus dorsi musculocutaneous flap combined with liposuction). RESULTS: After the treatment, the upper limb perimeter reduced in varied degrees. Nuclear lymphatic radiography showed notable changes in lymphatic circulation. The effective results were steady during the follow-up of 3-18 months. CONCLUSION: Flap transplantation combined with liposuction is a useful treatment for limb lymphedema from radical mastectomy.