Objective To investigate the proteomic changes among normal skin tissues in young and elderly people and cutaneous squamous cell carcinoma (cSCC) tissues in elderly patients with cSCC, find proteins associated with skin aging and cSCC, and provide a new basis for target screening for cSCC therapy. Methods Five cSCC tissue samples from 5 elderly patients with cSCC and 10 normal skin tissue samples from 5 young and 5 elderly people removed during surgery between January 2019 and December 2020 in West China Hospital of Sichuan University were selected. The differences in tissue morphology and structure were observed by hematoxylin-eosin staining, and the whole protein group was qualitatively and quantitatively analyzed by pressure cycle technique. Results With aging, the structure of skin tissues underwent corresponding changes, including thinning of the skin, increased collagen fiber density, and more organized arrangement. Compared to normal skin tissue, cSCC tissue exhibited epithelial cell dysplasia, atypical mitoses, nest-like distribution of cancer cells, infiltration of inflammatory cells, and formation of keratin pearls. Proteomic analysis identified 3008 specific proteins, and there were 37 proteins with common differential expression. Further screening of databases identified 8 proteins derived from the extracellular matrix, primarily involved in morphological structure formation, tensile strength, interaction with platelet-derived growth factors and receptors, collagen degradation metabolism, and cell adhesion. Conclusions Aging leads to changes in skin structure. The changes of tissue structure caused by aging lead to the weakening of skin barrier function. At the same time, aging leads to the down-regulated expression of protein with the function of inhibiting tumor progression and the up-regulated expression of protein with the function of promoting tumor progression in extracellular matrix. These changes may affect the occurrence and development of cSCC by affecting the regulation mechanism of tumor extracellular microenvironment.
Objective To detect the expression of heat shock protein 47 mRNA in pathological scar tissue by using real-time fluorescent quantitative reversetranscription-polymerase chain reaction (RT-PCR). Methods The tissues of normal skin(n=6), hypertrophic scar(n=6) and keloid(n=6) were adopted, which were diagnosised by Pathology Department. Based on fluorescent TaqMan methodology, the real-time fluorescent quantitative RT-PCR were adopted to detect the expression ofheat shock protein 47 mRNA. Results Compared with normal skin tissue（0.019±0.021）×105, the expressions of heat shock protein47 cDNA of hypertrophic scar tissue（1.233±1.039）×105 and keloid tissue（1.222±0.707）×105 were higher， being significant differences（Plt;0.05）. Conclusion A fluorescent quantitative method was successfully applied to detecting the expression of heat shock protein 47 mRNA. Heat shock protein 47 may play an important role in promoting the formation of pathological scar tissue.
Objective To investigate the effectiveness of nasolabial flap and ear cartilage in repairing defects after nasal ala basal cell carcinoma resection. Methods Between January 2012 and August 2014, 8 patients with nasal ala basal cell carcinoma underwent tumor resection and defect repair with nasolabial flap and ear cartilage. Among the 8 patients, 5 were male and 3 were female, with an average age of 65 years (range, 45-76 years). The left side and right side were involved in 3 cases and 5 cases respectively. Carcinoma confirmed by pathological examination in all patients. The time between first biopsy and resection was 7-14 days (mean, 10 days). The defect ranged from 1.5 cm×1.5 cm to 2.0 cm×1.5 cm after tumor resection, and the size of nasolabial flaps ranged from 4.0 cm×1.5 cm to 5.0 cm×2.0 cm. The operations of cutting off the pedicle and thinning skin flap were performed at 6 months after first operation. Results All flaps survived. Incisions healed by first intention, and no related complication occurred. No carcinoma recurred after cutting off the pedicle. All patients were followed up for 6 months. All patients were satisfied with the nasal contour, symmetrical projection of the alar dome, and no obvious scar. Conclusion Nasolabial flap transfer and ear cartilage transplant method not only can repair the nasal ala defects, but also can avoid obvious scar and obtain good nasal ala contour profile. The shortcoming is that patients have to receive two operations.
ObjectiveTo investigate the expression and significance of peroxisome proliferator activated receptor γ(PPAR-γ) in human keloid. MethodsTwenty-three keloid samples were harvested from the patients undergoing keloid and auto-skin grafting operation as the experimental group (keloid group), and the residual normal skin after auto-skin grafting operation was collected as the control group. The expression of PPAR-γ protein was examined by immunohistochemistry staining in both keloid and normal skin. Referring to Shimizu immunohistochemical standard, the result was graded; the positive rate of samples and the rate of positive cells were calculated. ResultsImmunohistochemistry staining showed that PPAR-γ protein was expressed in both keloid and normal skin. In keloid, it located in the pricle cell layer, and granular layer of epidermis, and the dermal vessel; the degree of dyeing was very light. However, in normal skin, it located in the base layer of epidermis, dermal vessel walls, sweat glands and sebaceous glands; the dyeing degree was deeper. Immunohistochemical staining score in the keloid group (2.65±0.78) was significantly lower than that in the control group (3.65±1.19) (t=5.030, P=0.000). The positive rate of samples in the keloid group (52.17%, 12/23) was significantly lower than that in the control group (82.61%, 19/23) (χ2=4.847, P=0.028). The rate of positive cells was 46.04%±8.61% in the keloid group, which was significantly lower than that in the control group (59.39%±11.26%) (t=5.974, P=0.000). ConclusionCompared with normal skin, the expression of PPAR-γ protein in keloid is down-regulated in in human keloid, indicating that PPAR-γ may be related to the formation of keloid.
Objective To investigate the possibility of enhancing the inducing rate of adipose-derived stem cells (ASCs) into epidermal cells in the medium containing all-trans retinoic acid (ATRA) by supplementing with HaCaT condition medium. Methods ASCs were isolated and identified by detecting the expression of CD34, CD45, CD73, CD90, and CD105 with flow cytometry and differentiating into adipose and osteoblast lineage in the induction medium. The air-liquid interface cell culture model was established with the Transwell Room. The induction medium A contained ATRA, epidermal growth factor (EGF), and keratinocyte growth factor (KGF), while the induction medium B contained ATRA, EGF, KGF, and HaCaT condition medium. Experiment was divided into three groups cultured for 12 days: induction medium A (group A), induction medium B (group B), basic medium (group C). The epidermal cell surface markers: cytokeratin (CK) 14, 15, 16, 19 (Pan-CK) were detected by flow cytometry and CK14 were identified by immunofluorescence stain. Results After induction for 12 days, flow cytometry showed that the positive rate of Pan-CK in group B [(22.0±3.5)%] was higher than that in group A [(11.9±2.7)%], which were both higher than that in group C [(1.1±0.3)%], and the differences were statistical significantly (P<0.01). Immunofluorescence stain showed that the positive rate of CK14 in group B was higher than that in group A [(19.5±7.0)%vs. (10.8±5.7)%, P<0.01], and the expression of CK14 was negative in group C. Conclusion HaCaT condition medium can enhance the ability of ASCs differentiation into epidermal cells in the culture medium containing ATRA.
Objective To evaluate the effectiveness and advantages of the wide local excision for Paget’s disease involing the penis and scrotum by comparing with the radical excision. Methods A retrospective analysis was made on the clinical data of 41 patients with Paget’s disease involving penis and scrotum who met the inclusion criteria between November 2010 and August 2015. Among them, 14 patients received wide local excision (group A), and 27 patients received radical excision (group B). No significant difference was found in age, course of disease, and lesion site between two groups (P>0.05). The recurrence rate, operative time, times of intraoperative frozen section pathology, hospitalization time, grade of wound healing, appearance and functions satisfaction were recorded and compared between two groups. Results The operative time and hospitalization time in group A were significantly shorter than those in group B (P<0.05); the times of intraoperative frozen section pathology in group A were significantly less than that in group B (P<0.05). All patients were followed up 13 to 67 months (mean, 35.5 months) in group A and 11 to 70 months (mean, 38.8 months) in group B. Grades A, B, and C wound healing was obtained in 11 cases, 2 cases, and 1 case of group A and in 12 cases, 7 cases, and 8 cases of group B respectively, showing significant difference between two groups (Z=–2.102, P=0.036). The 5-year recurrence rate was 28.6% (4/14) in group A and 22.2% (6/27) in group B, showing no significant difference (χ2=0.202, P=0.654). The score of satisfaction in appearance and functions in group A was significantly higher than that in group B (t=–2.810, P=0.008). Conclusion Paget’s disease involving penis and scrotum has a slow disease progression and good prognosis. Wide local excision can relieve symptoms effectively and obviously decrease perioperative risk in elderly patients, with no significant increase of the recurrence rate.
【摘要】 目的 建立兔耳中期瘢痕动物模型，寻找兔耳瘢痕形成的最佳位点。 方法 选用日本大耳白兔20只，在兔耳腹侧选定6个位点，作直径1 cm直达软骨表面的皮肤全层及软组织缺损240个。创面暴露，于伤后7 d去除软骨上面的肉芽及血浆痂壳一次。术后连续3个月观察创面自然愈合及瘢痕增生情况;用HE及苦味酸-天狼星红染色观察瘢痕形成及胶原分布情况;用计算机图像分析系统测定胶原含量。 结果 兔耳腹侧可制作类似人的增生性瘢痕模型，瘢痕的发生率42.5%~56.7%，瘢痕增生的高峰在造创后30~50 d。不同位点瘢痕增生程度不同，胶原含量也不同。 结论 兔耳腹侧可建立中期瘢痕动物模型，兔耳腹侧的中分和耳尖外侧部分是制作兔耳增生性瘢痕的理想位点。【Abstract】 Objective To establish an animal model of intermediate stage hypertrophic scar on the rabbit ears and to find out the best sites of scar formation. Methods A total of 240 full-thickness skin and tissue defect directing access to the cartilage surface was created on the ventral side in 20 Japan white rabbits and each ear contain 6 defect sites.The wound was treated by exposure method.On the 7th day after operation, the granulation tissue and plasma shell were removed on the cartilage.Wound healing and scar proliferation under natural condition were observed continuously for 3 months.The scar formation and collagen distribution were observed by HE and Sirius red staining, and the collagen content was analyzed by using computer image analysis system. Results The ventral wound of rabbit’s ears produced hypertrophic scar similar to human hypertrophic scar, the incidence of scar was between 42.5% to 56.7%.The peak of scar proliferation was in 30 days to 50 days after operation.The degree of scar proliferation and collagen content varied at different sites. Conclusion The ventral wound of rabbit’s ears can produce intermediate stage hypertrophic scar model, the middle sites and the lateral ear tip are ideal site for madding animal model of hypertrophic scar on the rabbit ears.