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 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 observe the protein expression of c-Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) in normal skin and keloid and to explore their influences on the formation of kloid. Methods Keloid tissues and normal skin tissues were collected from 16 keloid resection patients (experimental group) and 10 voluntary plastic surgery patients (control group). In the experimental group, the keloid formation time ranged from 8 months to 10 years; the keloid tissues were collected from the chest in 6 cases, the ear lobe in 4 cases, the perineum in 2 cases, the shoulder in 3 cases, and the abdomen in 1 case; and all keloid tissues were confirmed by pathological examination. In the control group, normal skin tissues were collected from the abdomen in 4 cases, the thighs in 3 cases, the shoulder in 2 cases, and the back in 1 case. Two-step l ine of Envision immunohistochemical staining was performed to observe the expressions of nonphosphorylated and phosphorylated JNK and ERK; Image Pro Plus 4.5 image analysis system was used to measure the integrated absorbance (IA) and to observe the positive staining strength. Results The immunohistochemical staining showed that no obvious expressions of phosphorylated and non-phosphorylated ERK, JNK were observed in the fibroblasts of the control group, and the expressions of phosphorylated JNK and ERK proteins were significantly higher in the experimental group than in the control group (P lt; 0.05). There was no significant difference in the expressions of non-phosphorylated JNK and ERK proteins between 2 groups (P gt; 0.05). Conclusion Activation of ERK and JNK pathways might be involved in formation of keloid.
Objective To investigate the operative method and efficacy of subcutaneous pedicle scar-band rotation flap in the treatment of cervical postburn scar contracture. Methods Between August 2008 and May 2010, 15 patients with cervical postburn scar contracture were treated with subcutaneous pedicle scar-band rotation flaps, including 9 males and 6 females with an average age of 17.3 years (range, 7-35 years). The disease duration was 1-8 years (mean, 3 years). The locations were the left cervical region in 6 cases, the right cervical region in 8 cases, and mental cervical angle region in 1 case. According to LI Ao’s classification standard for cicartrical contracture, there were 12 cases of grade I and 3 cases of grade II. The area of scar ranged from 8 cm × 5 cm to 25 cm × 12 cm. After scar relaxation, wounds were repaired with the subcutaneous pedicle scar-band rotation flaps of 7 cm × 5 cm to 15 cm × 10 cm at size. In 3 cases of grade II, free split thickness skin grafts (7 cm × 4 cm to 12 cm × 7 cm at size) were used simultaneously. Results After 2 weeks of operation, 2 scar-band flaps had distal partial necrosis and healing was achieved after dressing change with formation of hypertrophic scar; the others survived with healing of incisions by first intention. After 6-12 months of follow-up, all patients possessed good cervical contours, sufficient release of scar contractures, and normal cervico-mandicular or mental cervical angles. The skin’s color and texture were satisfactory. There was no recurrence of cervical scar contracture and other complications. All patients acquired normal cervical movement at last follow-up. Conclusion Subcutaneous pedicle scar-band rotation flap is a simple, efficient, and versatile technique in release of cervical postburn scar contracture. It is an effective method to make use of the lateral excess scar flap to resurface defects caused by scar release.
【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.
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.
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 seek for a method of constructing the tissue microarray which contains keloid, skin around keloid, and normal skin. Methods The specimens were gained from patients of voluntary donation between March and May2009, including the tissues of keloid (27 cases), skin around keloid (13 cases), and normal skin (27 cases). The specimens were imbedded by paraffin as donor blocks. The traditional method of constructing the tissue microarray and section were modified according to the histological characteristics of the keloid and skin tissue and the experimental requirement. The tissue cores were drilled from donor blocks and attached securely on the adhesive platform which was prepared. The adhesive platform with tissue cores in situ was placed into an imbedding mold, which then was preheated briefly. Paraffin at approximately 70℃ was injected to fill the mold and then cooled to room temperature. Then HE staining, immunohistochemistry staining were performed and the results were observed by microscope. Results The constructed tissue microarray block contained 67 cores as designed and displayed smooth surface with no crack. All the cores distributed regularly, had no disintegration or manifest shift. HE staining of tissue microarray section showed that all cores had equal thickness, distinct layer, manifest contradistinction, well-defined edge, and consistent with original pathological diagnosis. Immunohistochemistry staining results demonstrated that all cores contained enough tissue dose to apply group comparison. However, in tissue microarray which was made as traditional method, many cores missed and a few cores shifted obviously. Conclusion Applying modified method can successfully construct tissue microarray which is composed of keloid, skin around keloid, and normal skin. This tissue microarray will become an effective tool of researching the pathogenesis of keloid.
Objective To explore the therapeutic effect of micro-autologous fat transplantation (MAFT) based on anatomical sub-unit principle of mid-face deep fat tissue in mid-face volumetric augmentation operation. Methods From August 2014 to February 2016, 46 patients suffering volumetric defect of mid-face were divided into the research group and the control group with 23 in each. The MAFT method was used in the two groups. Patients in the research group underwent fat transplatation based on anatomical sub-unit principle of mid-face deep fat tissue, of whom 16 accepted fat transplantaion in suborbicularis oculi fat area, 23 in deep media cheek fat area and 13 in buccal fat area. Patients in the control group accept conventional fat transplantation method; the fat granule were uniformly transplanted to the deep and superficial mid-face fat tissue. The curative effiency and patients' satisfaction were compared between the two groups. Results The curative effiency and patients' satisfaction of the research group (91.3%, 82.6%) were better than those of the control group (60.9%, 43.5%), and there were significant differences (P<0.05). All patients were followed up for 6 months to 2 years, with an average of 1 year. No complication occurred in both groups. Conclusion The MAFT operation based on anatomical sub-unit principle of mid-face deep fat tissue could precisely perform mid-face volumetric augmentation and rejuvenation.
ObjectiveTo develop a standardized venous thromboembolism (VTE) prevention program for burn patients and verify its safety and effectiveness by comparing with traditional thrombus prophylaxis.MethodsAll burn patients admitted and met selection criteria betweem April 2017 and September 2018 were included. Patients between January 2018 and September 2018 were included as the interventional group to implement standardized VTE prevention programs, while patients between April 2017 and December 2017 were included as the control group and traditional active and passive exercises were used to prevent VTE. There was no significant difference in the age, gender, ethnic group, marriage, education, occupation, type and site of the injury, burn area, operation time, and hospital stay between the two groups (P>0.05), which was comparable. The incidence of VTE, number of cases of tissue or organ hemorrhage, survival rate of skin grafting, and time of wound healing were compared.ResultsThe incidence of VTE was obviously lower in the interventional group (1.56%, 1/64) than in the control group (10.17%, 6/59) (χ2=−2.05, P=0.04). No bleeding occurred in any tissue or organ in the two groups. The survival rate of skin grafting and the time of wound healing were 89.06% (57/64) and (11.78±3.08) days respectively in the interventional group and 91.53% (54/59) and (11.66±2.30) days respectively in the control group; and the differences between the two groups were not statistically significant (χ2=0.21, P=0.65; t=−0.22, P=0.83).ConclusionThe standardized VTE prevention program can effectively prevent the occurrence of VTE, and its safety is relatively high.