晚期糖基化终末产物及其受体在瘢痕疙瘩中的表达

目的 研究晚期糖基化终末产物（AGE）及其受体（AGER）在瘢痕疙瘩中的表达。方法 瘢痕疙瘩、增生性瘢痕和正常人群血清、皮肤组织标本各20份,以荧光分光光度计检测三组人群血清中AGE含量,分别采用免疫组化法、Western印迹分析检测三组人群皮肤组织标本中AGE和AGER表达情况。结果 瘢痕疙瘩组血清中AGE含量为（0.713 ± 0.098） AU/ml,增生性瘢痕组为（0.699 ± 0.077） AU/ml,明显高于正常人群组（0.179 ± 0.056） AU/ml,三组差异有统计学意义（F = 283.82,P < 0.01）。免疫组化显示,AGE、AGER在瘢痕疙瘩、增生性瘢痕皮肤组织标本中表达阳性,正常人群组织表达则为阴性。Western印迹检测显示,瘢痕疙瘩、增生性瘢痕组织中AGE和AGER蛋白表达均明显高于正常人群,差异有统计学意义（F值分别为18.04、42.80,P < 0.05）,而瘢痕疙瘩和增生性瘢痕组之间差异无统计学意义（P > 0.05）。结论 AGE和AGER在瘢痕疙瘩中表达升高,在其发病过程中可能发挥一定的促进作用。     

Rising levels of serum S100 protein precede other evidence of disease progression in patients with malignant melanoma

BACKGROUND: Several serum markers may be useful in the detection of metastatic melanoma, but none is in routine clinical use. OBJECTIVE: To assess the validity of S100 protein as a serum marker of melanoma progression. METHODS: Serum S100 protein levels were measured in 496 serum samples from 214 melanoma patients, using the Sangtec luminescence immunoassay. There were 75 patients with stage 1 melanoma, 66 initially with stage 2 melanoma, 49 initially with stage 3 melanoma and 24 with stage 4 melanoma. RESULTS: Serum S100 protein levels were < 0.2 microg L-1 in 71 of 75 (95%) stage 1 patients. One patient who had a normal level developed local recurrence. Fifty-eight of 66 (88%) stage 2 patients also had normal serum S100 protein levels. One with elevated levels progressed to stage 3 melanoma and five with elevated levels progressed to stage 4 disease. The remaining two with elevated serum S100 protein remained well. Thirty-five of 49 (71%) stage 3 patients had normal levels and, of these, two have progressed to stage 4 disease. Three patients with stage 3 disease had an elevated serum S100 protein level on one occasion but remained well. Eleven of 13 patients who developed stage 4 melanoma during the study had rising levels of serum S100 protein > 0.2 microg L-1 5-23 weeks before detection of melanoma progression by conventional means. Twenty-two of 24 patients with stage 4 disease throughout the study had consistently elevated serum S100 protein levels, and the two patients with normal levels were clinically disease free after surgery and chemotherapy. None of 14 control subjects with atypical naevi had elevated S100 protein levels, and only one of 11 healthy normal controls had an elevated level. CONCLUSIONS: Thus, rising levels of serum S100 protein are a specific and sensitive clinically relevant marker of tumour progression in melanoma patients, which precedes other evidence of melanoma recurrence.     

In vivo quantitative analysis of advanced glycation end products in atopic dermatitis—Possible culprit for the comorbidities?

Advanced glycation end products (AGEs) interact with the membrane-bound receptor for AGEs (RAGE), consequently amplifying the inflammatory response. Soluble receptor for AGE (sRAGE) and endogenous secretory RAGE (esRAGE) act as decoys for AGE and competitively sequester RAGE ligands, thereby serving a cytoprotective role. Our objective was to investigate AGE expression and their receptors in the serum and skin of patients with atopic dermatitis (AD). In this case-control study, the levels of AGE, sRAGE and esRAGE were measured in the blood samples and corneocytes of 29 adult patients with AD and 12 healthy controls by ELISA. Corneocyte AGE levels increased in the AD group (P = .002). Higher corneocyte AGE levels were observed in the severe AD than in the milder form of AD. No significant difference in serum AGE level was observed in patients with AD and healthy controls. Serum sRAGE markedly decreased in patients with AD (P = .007) and serum esRAGE followed a similar trend. In conclusion, dermal accumulation of AGE in AD may have a role in fuelling skin inflammation. The potential after-effects of reduced neutralizer on systemic risk need further evaluation.     

Serum S100 concentrations are not useful in predicting micrometastatic disease in cutaneous malignant melanoma

BACKGROUND: S100 protein is an acidic calcium binding protein that is expressed by melanoma cells. Elevated serum values of S100 have been described in metastatic disease and it has been suggested that it may be used as an adjunct to staging and monitoring of treatment. Micrometastatic disease in the sentinel lymph node can be demonstrated by sentinel node biopsy (SNB) and the sentinel node status is known to be the most important predictor of relapse. OBJECTIVES: To determine whether serum S100 concentrations could predict the presence of micrometastatic disease. METHODS: Thirty-one patients with primary cutaneous melanoma > 1 mm were recruited from referrals to the Melanoma clinic. All patients had serum S100 concentrations evaluated prior to undergoing SNB. Serum S100 concentrations were established using an immunoluminometric method. Sentinel nodes were identified using a dual technique with both radiolabelled colloid (residual from preoperative lymphoscintigraphy) and blue dye according to the MD Anderson Cancer Center protocol. Results Nine of these 31 patients had evidence of micrometastatic disease on SNB. The mean serum S100 concentration of those with positive SNBs was 0.027 microg L-1 compared with 0.045 microg x L(-1) in patients with negative SNBs (normal < 0.14 microg x L(-1)). No patient in the study demonstrated raised concentrations of serum S100. CONCLUSIONS: We conclude that serum S100 concentrations do not predict the presence of micrometastatic melanoma in sentinel nodes in primary cutaneous melanoma.     

组织蛋白酶D调控皮肤成纤维细胞降解晚期糖基化终末产物的研究

目的 探究组织蛋白酶D（CatD）对皮肤成纤维细胞降解吞入胞内的晚期糖基化终末产物（AGE）的调控作用。方法 分别以CatB + CatL、CatD以及20S蛋白酶体的酶活性抑制剂CA074Me、天冬氨酸酶抑制剂（pepstatin A）、MG-132处理皮肤成纤维细胞,CCK8和荧光法检测细胞活性及蛋白酶活性。CA074Me、pepstatin A、MG-132组分别加入AGE-BSA孵育8 h,而对照组加入磷酸盐缓冲液（PBS）。孵育8 h后,置换含相应抑制剂的新鲜培养基继续培养24 h,流式细胞仪检测各时间点胞内AGE-BSA平均荧光强度。以不同浓度pepstatin A（37.5、75、150 μmol/L）或PBS处理皮肤成纤维细胞,加入AGE-BSA孵育,方法同前,ELISA检测各时间点胞内AGE-BSA平均浓度,计算降解率。慢病毒转染皮肤成纤维细胞,设空白对照组、空载病毒转染组、CatD高表达慢病毒转染组,荧光显微镜观察,RT-PCR、Western印迹法和荧光法检测各组CatD mRNA和蛋白表达及酶活性。分别加入AGE-BSA孵育,处理及检测方法同前,计算降解率。结果 1 μmol/L CA074Me组、75 μmol/L pepstatin A组及1 μmol/L MG-132组细胞增殖活性均在90%以上,与对照组（100%）差异无统计学意义（F = 1.525,P > 0.05）。去除AGE-BSA 24 h后,CA074Me + AGE-BSA组和MG-132 + AGE-BSA组胞内AGE-BSA荧光强度（275.00 ± 10.15、259.00 ± 11.14）均较其相应8 h点荧光强度（295.00 ± 6.56、285.67 ± 8.74）显著下降（配对t值分别为4.778、6.154,均P < 0.05）,而pepstatin A + AGE-BSA组8 h和32 h点细胞内AGE-BSA荧光强度差异无统计学意义（均P > 0.05）。37.5、75和150 μmol/L pepstatin A组24 h内对吞入胞内AGE-BSA的降解率分别为9.64% ± 1.27%、5.62% ± 0.47%和3.21% ± 0.73%,各组间差异有统计学意义（F = 45.876,P < 0.05）,且该降解率随pepstatin A浓度增加而降低（P < 0.05）。荧光显微镜下观察,空白对照组细胞未见荧光,空载病毒转染组和CatD高表达慢病毒转染组细胞荧光阳性率均在80%以上。CatD高表达慢病毒转染组CatD mRNA、蛋白表达及活性均较另2组显著上调（均P < 0.05）。CatD高表达慢病毒转染 + AGE-BSA组24 h内AGE-BSA的降解率显著高于AGE-BSA组和空载病毒转染 + AGE-BSA组（均P < 0.05）。结论 CatD可促进皮肤成纤维细胞降解吞入胞内的AGE-BSA。     

Predictive value of serum S100B for monitoring patients with metastatic melanoma during chemotherapy and/or immunotherapy

In the immunohistology of malignant melanoma the use of polyclonal antibodies against protein S100 is well established. Recently, it was shown that S100B, a subunit of the S100 protein family, is detectable in the serum of melanoma patients and correlates with the stage of the disease in patients with metastatic melanoma. In the present study, the first evaluation of a large number of treatment observations (n = 77) in 64 different patients during chemotherapy and/or immunotherapy for advanced metastatic melanoma (stage IV) is presented. All patients received treatment according to standardized protocols comprising 8 weeks of treatment followed by routine staging procedures to evaluate therapeutic outcome. In 13 patients with tumour enlargement after first-line therapy, a second-line treatment was subsequently given. S100B immunoradiometric assay (IRMA) tests were performed before, during and after treatment at scheduled time points. In the interim analysis at 4 weeks 29 of 37 (78%) patients with tumour progression during treatment showed a raised S100B level. In the final analysis at 8 weeks, 31 of these 37 patients (84%) demonstrated rising S100B values (P < 0.001). Patients who responded to treatment (stable or regressing metastatic disease) showed constant or declining S100B levels in 38 of 40 patients (95%) at the interim analysis, at 8 weeks this was further increased to 39 of 40 patients (98%; P < 0.001). Thus, the use of S100B for monitoring treatment is adequate in the majority of cases. Our observations are of great interest for therapeutic trials of adjuvant and palliative therapies as the rise of S100B levels might indicate that re-staging and/or changes in therapy strategies should be chosen.     

晚期糖基化终末产物对UVA照射皮肤成纤维细胞组织蛋白酶D表达及其活性的影响

目的：研究晚期糖基化终末产物（AGE）对长波紫外线（UVA）照射皮肤成纤维细胞组织蛋白酶D （CatD）表达和活性的影响。方法原代培养来自儿童包皮成纤维细胞。CCK?8法筛选对成纤维细胞无细胞毒性的AGE?牛血清白蛋白（BSA）浓度。分别以50、100、200 mg/L AGE?BSA孵育细胞24 h,以未处理细胞作为对照组,采用RT?PCR、Western印迹及荧光法检测AGE?BSA对细胞CatD表达和活性影响。将部分成纤维细胞分为6组,即对照组（正常皮肤成纤维细胞,不接受任何处理）、AGE?BSA组、BSA组、UVA组、UVA?AGE?BSA组、UVA?BSA组。AGE?BSA组加入最大无细胞毒性浓度AGE?BSA孵育24 h,BSA组加入相同浓度BSA孵育24 h,后3组除分别接受上述处理外再接受10 J/cm2 UVA照射。处理结束后,收集细胞mRNA和蛋白,采用RT?PCR、Western印迹及荧光法检测细胞CatD表达和活性。结果50、100、200 mg/L AGE?BSA对皮肤成纤维细胞增殖活性无显著影响。50 mg/L组、100 mg/L组、200 mg/L组细胞CatD mRNA水平分别为0.267±0.007、0.348±0.007、0.418±0.006,CatD蛋白水平分别为1.403±0.181、2.233±0.090、2.477±0.111,CatD活性分别为1.760±0.080、2.330±0.060、2.890±0.080,较相应对照组CatD mRNA（0.161±0.006）、CatD蛋白（0.903±0.200）以及CatD活性水平（1.100±0.090）均显著升高,均P<0.05。AGE?BSA呈剂量依赖性地刺激细胞CatD表达和活性。对照组、UVA组和UVA?AGE?BSA组细胞中CatD mRNA表达水平分别为0.155±0.005、0.480±0.005、0.394±0.008,蛋白表达水平分别为0.920±0.235、2.583±0.199、2.070±0.125,CatD活性分别为1.110±0.040、2.970±0.110、2.560±0.060;UVA组CatD mRNA水平、蛋白表达水平、酶活性均明显高于对照组（P<0.05）,但UVA?AGE?BSA组3种指标水平均显著低于UVA组（P<0.05）。结论 AGE可升高未接受UVA照射的皮肤成纤维细胞CatD表达和活性,但AGE却抑制UVA照射上调的CatD表达和酶活性。     

皮肤组织蛋白酶D表达与晚期糖基化终末产物堆积的相关性研究

目的 研究组织蛋白酶D（CatD）与晚期糖基化终末产物（AGE）在不同年龄、不同曝光程度皮肤中的表达及其相关性,初步探讨CatD在光老化皮肤AGE降解和堆积中的作用。方法 15 ~ 20岁、35 ~ 40岁、55 ~ 60岁及75 ~ 80岁患者曝光和非曝光部位皮肤组织,共8组,每组6份。采用免疫组化和免疫荧光双染法分别检测CatD和AGE在各组皮肤中的表达。采用析因设计方差分析、Wilcoxon秩和检验和Kruskal?Wallis秩和检验分析CatD和AGE表达与年龄、曝光的关系,并用Pearson相关系数分析CatD和AGE两者表达的相关性。结果 免疫组化显示,CatD表达随年龄增加而明显下降,而AGE的沉积则随年龄增加而逐渐增多;曝光部位CatD表达较同年龄组非曝光部位明显降低,而AGE沉积比同年龄组非曝光部位明显升高。析因设计方差分析显示,曝光会降低CatD的表达（F = 58.70,P < 0.001）,但会增加AGE的表达（F = 158.18,P < 0.001）。年龄的增长亦会引起CatD表达降低（F = 79.49,P < 0.001）,但AGE表达随年龄的增长而增加（F = 106.06,P < 0.001）。除了15 ~ 20岁年龄组,其他年龄组曝光组和非曝光组间CatD（35 ~ 40岁组：0.020 ± 0.005比0.032 ± 0.005;55 ~ 60岁组：0.012 ± 0.004比0.026 ± 0.002;75 ~ 80岁组：0.002 ± 0.001比0.013 ± 0.004）和AGE平均吸光度值（35 ~ 40岁组：0.030 ± 0.008比0.010 ± 0.003 ;55 ~ 60岁组：0.066 ± 0.010比0.021 ± 0.004 ;75 ~ 80岁组：0.085 ± 0.015比0.035 ± 0.009 ）差异均有统计学意义（均P < 0.001）。在固定曝光因素各水平条件下,不同年龄组间CatD和AGE表达差异亦均有统计学意义（均P < 0.001）。免疫荧光双染结果与免疫组化结果相似。Pearson相关分析显示,曝光部位皮肤CatD表达与AGE沉积呈高度负相关（r = -0.915,P < 0.05）,在非曝光皮肤中两者呈中度负相关（r = -0.730,P < 0.05）。结论 随年龄增长皮肤组织中CatD表达水平下降,而AGE表达水平上升。非曝光部位皮肤CatD和AGE表达呈中度负相关,曝光部位皮肤CatD和AGE表达呈高度负相关,CatD很可能在光老化皮肤AGE降解和堆积中起重要作用。     

S100A7 (psoriasin) inhibits human epidermal differentiation by enhanced IL‐6 secretion through IκB/NF‐κB signalling

Psoriasin (S100A7), a member of the S100 protein family, is a well‐known antimicrobial peptide and a signalling molecule which regulates cellular function and is highly expressed in hyperproliferative skin conditions such as atopic dermatitis (AD) and psoriasis with disrupted skin barrier function. However, its role in epidermal differentiation remains unknown. We examined the effect of S100A7 on epidermal differentiation in normal human keratinocytes (NHKs) and on a reconstituted human epidermis model. When NHKs were exposed to disruptive stimuli such as Staphylococcus aureus, ultraviolet irradiation and retinoic acid, the secretion of S100A7 into the culture medium increased and the expression of epidermal differentiation markers decreased. Treatment of NHKs with S100A7 significantly inhibited epidermal differentiation by reducing the expression of keratin 1, keratin 10, involucrin and loricrin and by increasing the expression of abnormal differentiation markers (keratin 6 and keratin 16). We verified that the MyD88‐IκB/NF‐κB signal cascade was activated via RAGE after S100A7 treatment, resulting in the upregulation of interleukin‐6. Finally, we confirmed that S100A7 is a negative regulator of epidermal differentiation using a reconstituted human epidermis model. This study suggests that S100A7‐related signalling molecules could be potent targets for recovering skin barrier function in AD and psoriasis where S100A7 is accumulated excessively.     

Serum levels of leptin receptor in patients with malignant melanoma as a new tumor marker

Leptin is known to be abnormally expressed in a variety of cancers, and leptin receptors have been reported to be expressed on human melanoma cells. In this study, we evaluated the possibility that the serum levels of leptin receptor could be a tumor marker of malignant melanoma (MM). Serum samples were obtained from 71 patients with MM, and the serum levels of leptin receptor were measured by double‐determinant ELISA. Interestingly, serum levels of leptin receptor decreased gradually with the stages of MM, being highest at in situ and lowest at stage IV. There was also a trend of reverse correlation between tumor thickness and serum levels of leptin receptor. To our knowledge, this is the first report investigating the serum levels of leptin receptor in MM, and serum leptin receptor levels may be used as a useful tumor marker of MM.     

Dacarbazine, nimustine hydrochloride, cisplatin and tamoxifen combination chemotherapy for advanced malignant melanoma.

Melanoma is an uncommon disease in Japan. The incidence, however, has been gradually increasing in the last two decades, as in many other countries worldwide. Ten patients with metastatic malignant melanoma were treated between March of 1997 and April of 1998 in the Department of Dermatology, National Cancer Center Hospital, with a combination chemotherapy consisting of dacarbazine (DTIC), nimustine hydrochloride (ACNU), cisplatin (CDDP), and tamoxifen (TAM). The patients characteristics were as follows: four were males and six females; the age range was 33-70 years; all were Japanese; sites of primary disease: extremities 4, primary unknown 3, nasal cavity 1, anus 1, scalp 1; sites of metastases: lymph nodes 6, pulmonary system 5, skin 2, liver 3, gall bladder 1, adrenal gland 1. The chemotherapy regimen included DTIC 220 mg/m2/i.v. on days 1 through 3, ACNU 60 mg/m2/i.v. on day 1, cisplatin 25 mg/m2/i.v. on days 1 through 3, and tamoxifen 10 mg p.o. twice daily. One patient achieved a complete response and 3 showed partial responses. The response rate was 40%. The four responders included those with metastases to the nodes, lung, and liver. The main toxicities were nausea, vomiting, leucopenia, anemia, and thrombocytopenia. This regimen is a fairly effective combination against metastatic melanoma.     

寻常型银屑病患者及正常人S100A6、S100A8、S100A9基因的研究

目的：通过测定家族性寻常型银屑病患者和正常人中S100A6、S100A8和S100A9基因的外显子编码区序列，探索其与银屑病发病的关系。方法：从家族性寻常型银屑病患者和正常人的外周血中提取基因组DNA，用自动测序法测定S100A6、S100A8和S100A9基因的外显子编码区序列。结果：患者和正常对照组的S100A6、S100A8、S100A9基因外显子编码区序列均相同，未发现基因多态性。结论：S100A6、S100A8、S100A9基因的外显子编码区序列与家族性寻常型银屑病发病无相关性。     

皮肤肿瘤中的雌激素受体及皮肤恶性黑素瘤中雌激素和孕激素受体及C－erbB2癌基因的表达

通过对８个病种的２１例皮肤恶性肿瘤进行的雌激素受体的免疫组化标记（ＡＢＣ法），发现有２例皮肤恶性黑素瘤呈阳性表达。在１５例皮肤恶性黑素瘤中进行的免疫组化染色中（ＬＳＡＢ），雌激素受体阳性７例、孕激素受体阳性３例、Ｃ－ｅｒｂＢ２癌基因阳性７例。作者认为通过进一步研究有望获得恶性黑素瘤激素治疗和预后观察的生物学依据     

VEGF-C及其受体在皮肤恶性黑素瘤中的表达及其临床意义

目的：研究皮肤恶性黑素瘤（MM）组织中血管内皮生长因子C（vascular endothelial growth factor-C,VEGF-C）及其受体VEGFR-3（flt-4）的表达情况,分析其在肿瘤淋巴管的生长及淋巴转移中的作用。方法：采用免疫组化SP法分析29例MM石蜡标本中VEGF-C/flt-4蛋白的表达情况,并与20例色素痣石蜡标本中VEGF-C/flt-4蛋白表达情况相对比,统计分析其与临床病理特点之间的关系。结果：VEGF-C蛋白的阳性率为96.6%（28/29）,flt-4的阳性率为86.2%（25/29）,明显高于色素痣（P〈0.01）,与肿瘤的病理分级、临床分期和肿瘤的淋巴转移显著相关（P〈0.05）;VEGF-C和flt-4的阳性表达相一致。结论：VEGF-C/flt-4在皮肤恶性黑素瘤中的表达明显高于色素痣,与肿瘤的预后密切相关,在肿瘤淋巴转移中可能有重要作用。     

The risk for cutaneous malignant melanoma, melanoma in situ and intraocular malignant melanoma in relation to tobacco use and body mass index

BACKGROUND: The incidence of cutaneous malignant melanoma (CMM) and melanoma in situ (MIS) has been increasing during the last 50 years. Malignant melanoma (MM) is also the most common intraocular malignancy (IMM). Besides ultraviolet radiation, the cause of these tumours is largely unknown. OBJECTIVES: We designed a study to examine the effect of body mass index (BMI) and tobacco use on the risk for MM and MIS. METHODS: Analyses were performed on a nationwide cohort of 339 802 Swedish construction workers. Exposure information was collected prospectively by questionnaires combined with personal interviews. RESULTS: Follow up yielded a total of 7 663 400 person-years during which 1639 workers developed MM/MIS. The risk for MM/MIS was reduced in current or previous smokers compared with those who had never smoked, both when analysing all smoking tobacco products combined and when analysing cigarette and pipe smokers separately. The risk was further diminished with longer duration of smoking and greater quantity of tobacco smoked. The effect was more evident in CMM/MIS than in IMM. Snuff taking conferred a decreased risk for CMM/MIS, and a BMI over normal weight range conferred an increased risk for CMM. CONCLUSIONS: Tobacco smoking was found to be inversely associated with the risk for CMM and MIS. The mechanism of action is unknown but it has been suggested to be due to the immune suppressive effect that tobacco exerts which would be protective against deleterious immune reactions caused by, for example, the sun. Neither is the mechanism behind the higher risk for CMM due to being overweight known. One hypothesis is that it is an effect of a hormonal imbalance. Further studies are required to elucidate these mechanisms.     

Modeling clinical efficacy of the S1P receptor modulator ponesimod in psoriasis

Background

Ponesimod is currently the only S1P receptor modulator studied in psoriasis. In a dose-finding study, the active doses showed similar efficacy.

寻常型银屑病患者皮损中白介素-22和S100A7,A8,A9mRNA的表达及关系

目的研究白介素-22(IL-22)和S100A7,A8,A9mRNA在寻常型银屑病皮损中的表达及关系。方法采用逆转录聚合酶链反应(RT-PCR)技术对35例寻常型银屑病患者皮损和16例正常人皮肤中IL-22,S100A7,A8,A9mR-NA进行检测。结果①IL-22和S100A8,A9mRNA在寻常型银屑病皮损中呈阳性表达,而在正常皮肤中为阴性;S100A7mRNA在寻常型银屑病皮损中的表达水平为1.133±0.0403,较正常对照组0.744±0.0371明显升高,差异有显著性(P<0.01)。②IL-22mRNA与S100A7,A8,A9mRNA的表达呈显著正相关r1=0.543,r2=0.774,r3=0.621,P均<0.01。结论IL-22和S100A7,A8,A9与银屑病的发生和发展中明确相关,可能成为银屑病治疗新的靶位点。