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Objective:To investigate the effect of long non-coding RNA 068 (lncRNA 068) on the migration of a melanoma cell line A375, and to explore its mechanism of action.Methods:From December 2015 to November 2020, 21 patients with pathologically confirmed cutaneous melanoma were collected from Department of Dermatology, Affiliated Hospital of Nantong University, and quantitative PCR (qPCR) was performed to determine the expression of lncRNA 068 in melanoma and paracancerous tissues. LncRNA 068 was overexpressed or knocked down via lentiviral transfection in A375 human melanoma cells in the following experiments. Specifically, A375 cells were divided into lentiviral vector (LV) -green fluorescent protein (GFP) group and LV-lncRNA 068 group to be transfected with a GFP-expressing LV and a LV containing lncRNA 068 respectively in the overexpression experiment, and were divided into LV-LacZ short hairpin RNA (shRNA) group and LV-lncRNA 068 shRNA group to be transfected with a LV containing the reporter gene LacZ-specific shRNA and a LV containing the lncRNA 068-targeting shRNA respectively in the low-expression experiment, with the LV-GFP group and LV-LacZ shRNA group serving as the control groups. Transwell and scratch assays were performed to evaluate cell migration, EdU cell proliferation assay and cell counting kit-8 (CCK8) assay to determine the proportion of proliferative cells and cell viability respectively, and immunofluorescence staining was conducted to evaluate epithelial-mesenchymal transformation in the above groups. Lentivirus-transfected A375 cells from the above groups were inoculated into the axillae of BALB/c nude mice, and tumor volume was measured and calculated every 3 days. After 30 days, all mice were sacrificed, and tumor tissues were resected to measure the tumor volume and weight. Cultured B16F10 cells were subcutaneously inoculated into the back and foot of BALB/c nude mice to construct mouse models of subcutaneously transplanted B16F10 melanoma. After 2 weeks, the mice were sacrificed, and qPCR and Western blot analysis were performed to determine the mRNA expression of inflammatory factors in transplanted B16F10 melanoma and paracancerous tissues, and expression of IκB kinase (IKK) /P65 signaling pathway-related proteins, respectively. Comparisons between 2 groups were done by t test, and comparisons of tumor volume and weight at different time points among groups were done by repeated measures analysis of variance. Results:qPCR showed that the relative expression of lncRNA 068 was significantly lower in human melanoma tissues and transplanted B16F10 murine melanoma tissues (0.414 ± 0.109, 0.717 ± 0.041, respectively) than in the corresponding paracancerous tissues (1.050 ± 0.103, 1.011 ± 0.023, t = 19.48, 10.83, respectively, both P < 0.001). Transwell and scratch assays both showed that the cellular migratory ability was significantly lower in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.01), and significantly higher in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Immunofluorescence assay showed significantly increased fluorescence intensity of E-cadherin and decreased fluorescence intensity of N-cadherin in the LV-lncRNA 068 group compared with the LV-GFP group (both P < 0.001), but significantly decreased fluorescence intensity of E-cadherin and increased fluorescence intensity of N-cadherin in the LV-lncRNA 068 shRNA group compared with the LV-LacZ shRNA group (both P < 0.05). In vivo tumor formation experiment in nude mice showed that there were no significant differences in the volume or weight of melanoma between the LV-lncRNA 068 group and LV-GFP group (both P > 0.05), as well as between the LV-lncRNA 068 shRNA group and LV-LacZ shRNA group (both P > 0.05). As qPCR and Western blot analysis showed, the mRNA and protein expression of interleukin-10 (IL-10) and claudin-1 in A375 cells were significantly higher in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.05), but significantly lower in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Compared with the paracancerous tissues, B16F10 melanoma tissues showed significantly decreased mRNA expression of IL-10 ( P < 0.01), but significantly increased mRNA expression of IL-6 and tumor necrosis factor-α, as well as protein expression of phosphorylated P65 and phosphorylated IKK ( P < 0.01) . Conclusion:Overexpression of lncRNA 068 can inhibit the migration of A375 melanoma cells, and may affect the development of inflammation and inhibit the epithelial-mesenchymal transformation by inhibiting the IKK/P65 signaling pathway.
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Objective@#The study presents a new method to prefabricate the nasoalveolar molding appliances for preoperative cleft lip and palate by using three-dimensional technology.@*Methods@#A long term retrospective study of 40 cases of infants with unilateral cleft lip and palate who underwent the preoperative 3D models of alveolar bone acquisition, computer aided design for the rapid prototyping process, gypsum powder printing maxillary three-dimensional entity model and install the appliance for 3-4 months (or alveolar cleft<2 mm). Simultaneously, primary rhinoplasty can be done during cleft lip repair. All patients had clinic visits three times each month.@*Results@#Deformities of infants who underwent this treatment, were significantly improved. The alveolar cleft was significantly reduced (P<0.01), while bilateral alveolar tissue volume was significantly increased (P<0.01). Alveolar bone morphology was more symmetrical than before.@*Conclusions@#The digital design and manufacture system of appliances for preoperative cleft lip and palate can be successfully applied to the preoperative orthodontic treatment. The sequence treatment of cleft lip and palate becomes better improved. It can shorten the time duration and the numbers of clinic visit on the basis of ensuring the accuracy of preoperative orthodontic effect.
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Objective To investigate the efficacy of dermabrasion combined with aminolevulinic acid-based photodynamic therapy (ALA-PDT) for the treatment of nasal nodular basal cell carcinoma (nBCC).Methods Twentyfive patients who were pathologically diagnosed as nasal nBCC with lesion area ＞ 1 cm2 but no bone or cartilage involvement,were included in this study and treated with dermabrasion combined with ALA-PDT.At first,the part of tumor protruding outside the skin was removed by artherectomy,then dermabrasion was carried out.The wound surface was topically treated with 20％ aminolevulinic acid solution for 3-4 hours away from light immediately after surgery,then irradiated with LED light at a mean dose of 100 J/cm2 for 20 minutes.ALA-PDT was performed once a week for 3 consecutive weeks.The degree of and time required for wound healing were assessed,and tumor recurrence,cicatrization and appearance outcomes were observed during 1 year after surgery.Efficacy was assessed comprehensively.Results No postoperative wound infection occurred in these patients,and the average time for wound healing was (11.2 ± 1.3) days.During 1 year after the treatment,no recurrence was found,while cicatricial contracture developed in 1 case,mild proliferative scar in 3 cases,and depressed scar in 4 cases.All the patients were satisfied with the treatment outcomes,except 1 patient who was basically satisfied.Conclusions Dermabrasion combined with ALA-PDT is easy to operate with rapid wound healing,low postoperative recurrence rate and high degree of patient satisfaction,and is worthy of clinical promotion.
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A 70-year-old female patient presented with several cutaneous masses on the left neck and trunk for 40 years. Some masses were removed by surgical operation more than 10 years prior to the presentation, but recurred subsequently along with an increase in lesion number. Persistent dull pain emerged at the lesion sites 2 months prior to the presentation and the masses on the left neck ulcerated 10 days prior to the presentation. Histopathology showed tumor cell clumps in the dermis with benign eccrine spiradenoma components in the centre region and carcinomatous components in the periphery. The carcinomatous components included slightly atypical cells and hyaline degeneration. Immunohistochemically, carcinoembryonic antigen and epithelial membrane antigen were observed in the tumor tissues. The case was diagnosed as eccrine spiradenocarcinoma. The masses were surgically removed, but recurred 1 month later, increased in size and number and ulcerated 3 months later, and the patient died 6 months after the surgery.
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Objective To observe and compare the efficacy and safety of implantation of tissue expanders adjacent to or under keloid tissues for large keloids on anterior chest. Methods Between Mar 2006 and June 2009, a total of 17 patients with large keloid lesions on anterior chest received treatment with 21 tissue expanders,among which 12 were placed under the normal skin adjacent to keloids, and 9 were inserted under the keloid lesions. The scar size varied from 4.5 cm × 3.0 cm to 15.7 cm × 5.5 cm. The capacity was 70 to 400 ml for expanders adjacent to the keloid tissue, 80 to 500 mi for those beneath the keloid tissues. After tissue expansion for 6 to 8 weeks, the expander was removed and keloid lesions were resected followed by the repair of defect with expanded flaps. Further more, the patients received intraoperative local intradermal injection of betamethasone and postoperative superficial electron beam irradiation with divided doses of 7 Gy in 3 consecutive days within 1 week after the surgery. Follow-up varied from 12 to 50 months. Results Twenty expanders, except 1expander pocket which was removed ahead of time due to infection, were implanted successfully during the whole course of treatment. The main complication was expander exposure in 4 patients, including 1 expander adjacent to the keloids and 3 under keloid lesions, which showed no significant influence on secondary operation. Fifteen patients reported relief of symptoms and achieved satisfactory outcomes, while 2 patients, including 1 treated with expanders adjacent to the keloids and 1 with expanders under the keloid tissue, showed great suture tension and experienced delayed stitch removal followed by the recurrence of keloids after the operation.Conclusions The implantation of tissue expanders under the adjacent normal skin or keloid lesions is an ideal treatment option for large keloids on anterior chest. Regional suture tension is a direct contributor to the recurrence of keloid formation after surgical excision.
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Objective To retrospectively analyze the therapeutic effect of wide surgical excision combined with low-dose adjuvant interferon-alpha 2b on cutaneous malignant melanoma.Methods A total of 32 patients with cutaneous malignant melanoma received wide surgical excision after preoperative examination and staging.The excisions were performed with a margin measuring 1-2 cm from the visible lesions or biopsy scars.Surgical modalities included direct suture after excision(4 patients),dactylolysis or toe amputation(6 patients),free skin grafting(15 patients),random skin flap transfer(3 patients)and pedicle skin flap transfer(4 patients).Lymph nodes were selectively dissected in 9 patients with regional transfer of lymph nodes,and inguinal lymph nodes were cleared away in 2 patients.One week after the operation,patients received adjuvant therapy with subcutaneous injection of interferon-alpha 2b(3 million IU,thrice per week)for one to three years.Results Preoperative tumor staging revealed 21 cases of cutaneous malignant melanoma at stage Ⅱ,and 11 cases at stage Ⅲ.The excisions healed by the first stage in all the patients.Up to June 2011,2 patients had been lost to follow up,5 patients with stage Ⅲ melanoma had died.Survival was observed in all of the 4 patients receiving 1-year follow up,12 of 13 patients receiving 1-3 year follow up,5 of 7 patients receiving 3-5 year follow up,and 4 of 6 receiving 5-year follow up.Of the 25 surviving patients,regional lymph node metastasis was observed in 8 patients,which developed within 2 years after the operation in 2 patients.The adjuvant therapy with interferon-alpha 2b lasted 3 years in 8 patients,and more than 1 year in 11 patients.Side effects were mild.Conclusion Wide surgical excision plus low-dose interferon-alpha 2b is effective for the treatment of stage Ⅱ and Ⅲ cutaneous malignant melanoma with lower local recurrence and higher survival rate.
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Objective To study the expression of gravin in the keloid and to investigate its probable role in the pathogenesis of keloid. Methods Skin biopsy was performed and skin samples were obtained from 18 normal human controls and 25 patients with keloid. The mRNA expression of gravin in specimens was detected by real-time fluorescent RT-PCR, and double immunofluorescence analysis was used to investigate the distribution of gravin protein in skin tissues. Results As detected by real time PCR, the relative expression of gravin mRNA was remarkably reduced in comparison with normal skin (0.4565±0.1728 vs 0.0953±0.0664, P<0.01). Results of double staining indicated that, in normal skin, gravin was primarily distributed in the cytoplasm of fibroblasts; while in keloid, it was observed mainly in macrophages and sporadically in fibroblasts. Conclusions Compared with the normal skin, keloid shows a different expression intensity and distribution of gravin, which might contribute to the development of keloid by regulating the proliferation of fibroblasts and activation of macrophages.