石蜡包埋组织中麻风菌基因扩增试验初探

目的建立用石蜡包埋组织中麻风菌DNA进行基因扩增的试验.方法用Texpat试盒将石蜡切片脱蜡、破壁和释放麻风菌DNA并用100%乙醇沉淀纯化DNA,用引物RPOT(F)和RPUT (R)进行PCR.结果细菌指数大于1+的33份石蜡包埋标本,28份PCR阳性,细菌指数为0的标本PCR阴性.阴性对照呈阴性反应,阳性对照呈阳性反应,方法敏感度达0.04pgDNA水平.结论用本法可从福尔马林固定的石蜡包埋组织切片中释放麻风菌DNA,并获得满意基因扩增结果.     

巢式PCR检测石蜡包埋组织中着色霉、孢子丝菌和马尔尼菲青霉

目的 评价巢式PCR对不同深部真菌感染组织石蜡包埋标本检测的价值。方法 收集着色芽生菌病、孢子丝菌病、马尔尼菲青霉病及其他深部真菌病石蜡组织标本共44份,行组织病理观察并提取石蜡组织标本中DNA。使用针对着色霉、孢子丝菌及马尔尼菲青霉核糖体DNA特定区域的特异性巢式PCR引物,分别对所提取的真菌DNA进行扩增。分析巢式PCR对这3种病原真菌扩增的敏感性和特异性,并与组织病理检查方法比较。结果 20例着色芽生菌病组织蜡块中8例扩增阳性,10例孢子丝菌病组织蜡块中7例扩增阳性,10例马尔尼菲青霉病组织蜡块均扩增阳性,其余对照深部真菌病组织蜡块扩增均为阴性,巢式PCR检测3种真菌的敏感性分别为40%、70%和100%,特异性均达到100%。组织病理检查3种真菌的阳性率分别为95%、70%、80%。结论 巢式PCR扩增石蜡包埋组织中的真菌DNA是诊断深部真菌病的一种方法,尤其适用于诊断马尔尼菲青霉病。     

以S2-R2为引物PCR检测石蜡切片中的孢子丝菌

目的 探讨应用特异性引物PCR方法检测石蜡切片中孢子丝菌的可行性。方法 选取30份大连及10份长春地区临床疑诊孢子丝菌病的石蜡切片标本,采用改良的微波脱蜡、液氮研磨-CTAB破壁法提取DNA,以特异性引物S2-R2进行PCR扩增,与真菌培养结果进行比对。结果 30份大连地区标本中22份见阳性PCR扩增产物（73.33%）。真菌培养阳性的22份标本中20份PCR出现阳性扩增产物（91%）,真菌培养阴性的8份标本中2 份PCR出现阳性扩增产物。10份长春地区标本7份见阳性PCR扩增产物（70%）。结论 以S2-R2为引物的 PCR适用于孢子丝菌病石蜡切片中病原菌的检测。     

T细胞受体γ引物组合检测蕈样肉芽肿基因重排的研究北大核心CSCD

目的探讨BIOMED-2系统T细胞受体（TCR）Y引物组合在蕈样肉芽肿（MF）患者不同来源标本中TCRY基因重排检测中的价值。方法收集28例MF患者15份石蜡组织样本、14份新鲜皮损及18份全血组织样本,提取DNA,利用BIOMED-2系统TCR7引物组合进行TCRY基因重排检测,比较3组不同来源样本的检出率并进行统计学分析。用SPSSl3．0软件进行统计分析。各组之间阳性率采用r检验或Fisher确切概率法检验。结果15份石蜡包埋组织标本3份TCRY基因重排阳性,18份血液标本11份阳性,14份新鲜皮损标本12份阳性,3组之间阳性率差异有统计学意义（r=13．047,P〈0．01）,新鲜皮损TCRY基因重排阳性率显著高于石蜡组织。2011年的6例石蜡标本3份基因重排阳性,阳性率显著高于其余9例2011年之前的石蜡标本（Fisher精确概率法,P=0．044）,与14例新鲜组织标本TCRY基因重排阳性率（12／14）相比差异无统计学意义（Fisher精确概率法,P=0．131）。血液标本TcRY基因重排阳性率低于新鲜皮损,两组之间差异无统计学意义（r=2．358,P〉0．05）。结论BIOMED-2系统TCRY引物组合适用于检测MF患者不同组织样本的TCR基因重排,更适用于新鲜的组织标本。     

T细胞受体γ引物组合检测蕈样肉芽肿基因重排的研究

【摘要】 目的 探讨BIOMED-2系统T细胞受体（TCR）γ引物组合在蕈样肉芽肿（MF）患者不同来源标本中TCRγ基因重排检测中的价值。方法 收集28例MF患者15份石蜡组织样本、14份新鲜皮损及18份全血组织样本,提取DNA,利用BIOMED-2系统TCRγ引物组合进行TCRγ基因重排检测,比较3组不同来源样本的检出率并进行统计学分析。用SPSS13.0软件进行统计分析。各组之间阳性率采用χ2检验或Fisher确切概率法检验。结果 15份石蜡包埋组织标本3份TCRγ基因重排阳性,18份血液标本11份阳性,14份新鲜皮损标本12份阳性,3组之间阳性率差异有统计学意义（χ2 = 13.047,P < 0.01）,新鲜皮损TCRγ基因重排阳性率显著高于石蜡组织。2011年的6例石蜡标本3份基因重排阳性,阳性率显著高于其余9例2011年之前的石蜡标本（Fisher精确概率法,P = 0.044）,与14例新鲜组织标本TCRγ基因重排阳性率（12/14）相比差异无统计学意义（Fisher精确概率法,P = 0.131）。血液标本TCRγ基因重排阳性率低于新鲜皮损,两组之间差异无统计学意义（χ2 = 2.358,P > 0.05）。结论 BIOMED-2系统TCRγ引物组合适用于检测MF患者不同组织样本的TCR基因重排,更适用于新鲜的组织标本。 【关键词】 基因重排,γ链T细胞抗原受体; 肉芽肿,蕈样; 聚合酶链反应     

荆州地区2014～2016年艾滋病初筛的检测与分析

目的:探讨荆州地区2014~2016年艾滋病(AIDS)初筛检测情况,以更好了解本地区艾滋病传播情况,为预防和控制作参考。方法:选取2014年1月至2016年12月来自本院门诊及住院、个体诊所、妇科医院、乡镇医院(4所)、部分综合医院和兵检等525例病例为研究对象,对其中艾滋病抗体初筛阳性标本的初筛实验和确认实验结果进行分析。结果:2014~2016年检测数分别为99例、177例、249例。2014年初筛阳性率为10.1%,确认阳性率10%;2015年初筛阳性率11.9%,确认阳性率19.1%;2016年初筛阳性率15.7%,确认阳性率28.2%,2016年确认阳性率和其他两组比较明显偏高,比较差异显著(P<0.05)。2014~2016年双阳初筛阳性数为39例,一阴一阳数31例,确认阳性中双阳数为16例(41.0%),一阴一阳为0例,两者比较差异显著(P<0.05);70份初筛阳性标本S/CO均≥6初筛12例,确认阳性数11例,比率为91.7%;另外一种试剂实验2~6确认阳性率33.3%;另外一种试剂实验<2确认阳性率12.5%,两种试剂实验均≥6确认阳性率和其他比较差异显著(P<0.05)。结论:对临床上S/CO比值在6.0以上标本,要进行确认实验才能判定。     

实时荧光定量PCR检测石蜡标本中麻风菌DNA

目的：评价实时定量荧光PCR(Real-time PCR)法检测石蜡标本中麻风菌DNA的应用价值。方法：根据基因库(GenBank)发表的M. leprae的全基因组序列,以一段重复序列(repetitive ele￣ment,RLEP)为扩增靶序列,合成引物和探针,构建质粒pGEMT-101作为标准品,用Real-time PCR对石蜡标本进行麻风菌DNA检测,并评价其敏感性。结果：对52例麻风患者的石蜡包埋组织标本麻风菌进行了检测,不同型别麻风(LL：8;BL：10;BT：28;TT：6)的石蜡标本检测阳性率分别为100％(8/8)、80％(8/10)、78.57％(22/28)、50％(3/6)。结论： Real-time PCR方法可在石蜡标本中快速灵敏的检测到麻风菌DNA。     

女性尖锐湿疣和宫颈上皮内瘤变HPV16型E2、E6、E7基因突变及E6、E7抗原表达

目的研究女性尖锐湿疣(CA)和宫颈上皮内瘤变(CIN)中人乳头瘤病毒(HPV)亚型分布,以及高危型HPV16的E2、E6、E7基因突变及致癌抗原E6、E7表达。方法从24例女性CA、11例2级CIN(CIN2)和23例3级CIN(CIN3)共58份石蜡包埋组织中提取HPV基因组DNA,用PCR-膜杂交法检测HPV亚型,DNA测序法检测HPV16型E2、E6、E7基因突变,免疫组化S-P法检测HPV16型E6、E7抗原表达。结果女性CA、CIN2和CIN3患者58份标本中共检出10个HPV亚型,高危型HPV检出率分别为20.8%、45.5%和60.9%,HPV16型检出率分别为12.5%、18.2%和34.8%。24例女性CA中高危型和低危型HPV检出率为20.8%和70.8%,宫颈CA高危型HPV检出率60.0%。HPV16型E2、E6、E7基因分别检出10、4和4个突变位点。首次在1例女性外阴CA和3例CIN3中一致检出HPV16型E2基因9个突变位点。HPV16型E6抗原在CA、CIN2和CIN3均有表达,但E7抗原仅在CIN3表达。结论女性宫颈CA和CIN多为高危型HPV感染,女性外阴CA的HPV16型E2基因突变可能与CIN3发生有一定关联。     

子宫内膜取样器获取两种标本筛查子宫内膜病变的临床应用研究

目的评价子宫内膜取样器Pipelle获取两种子宫内膜标本筛查子宫内膜病变的临床应用价值。方法采用前瞻性、自身对照的方法,选取2020年12月至2022年3月梧州市工人医院收治的316例存在明确手术指征需进行宫腔镜检查和子宫内膜活检的患者作为研究对象。采用子宫内膜取样器Pipelle获取子宫内膜标本,随后进行宫腔镜检查+常规诊刮术,Pipelle获取的子宫内膜标本同时进行液基细胞学制片及细胞模块石蜡包埋切片,以宫腔镜检查+常规刮宫术的组织学病理作为金标准进行比较。结果(1)常规诊刮术行组织石蜡切片满意率为97.78%(309/316),Pipelle获取标本行液基细胞学制片满意率为93.99%(297/316),行细胞模块石蜡包埋切片满意率为98.40%(311/316),两种石蜡切片的取材满意率均高于液基细胞学制片的取材满意率,差异具有统计学意义(P<0.05);两种石蜡切片的取材满意率比较,差异无统计学意义(P>0.05);Pipelle获取标本时无需麻醉和扩宫,患者无疼痛感。(2)Pipelle获取细胞模块组织病理学与液基细胞学筛查和诊断子宫内膜病变的灵敏度分别为68.75%、40.63%,特异度分别为99.27%、98.10%,阳性预测值分别为91.67%、72.22%,阴性预测值分别为96.45%、93.14%,符合率分别为96.08%、91.86%,Youden指数分别为0.61、0.39,Kappa值分别为0.76、0.47。结论Pipelle获取子宫内膜标本满意率高,标本进行细胞模块包埋石蜡切片组织病理学检查的准确性高于液基细胞学,Pipelle取样操作简便,无需麻醉,无疼痛感,适用于基层医院筛查子宫内膜病变。     

阴茎尿道口内生殖器疣病人精液中的人类乳头瘤病毒DNA的检测

生殖道的人类乳头瘤病毒(HPV)感染是通过性传播的,有研究表明这种病毒感染与良性、恶性肿瘤前及恶性肿瘤损害有关.为了研究HPV传播的可能性,作者用Southern印迹杂交试验检测了阴茎尿道口内生殖器疣病人的HPV DNA.15份精液标本取自泌尿生殖科门诊10例未经治疗的患者阴茎尿道口内生殖器疣的病人,标本采集后放入灭菌的塑料器皿中.DNA提取和杂交试验:每份标本取0.4ml放入0.1%SDS溶液中用蛋白酶K(100μg/ml)消化,放置过夜.标本用等量的酚/氯仿混合液萃取DNA两次,然后再用氯仿/异戊     

Outcomes and adverse effects of ablative vs nonablative lasers for skin resurfacing: A systematic review of 1093 patients

It is generally believed that ablative laser therapies result in prolonged healing and greater adverse events when compared with nonablative lasers for skin resurfacing. To evaluate the efficacy of ablative laser use for skin resurfacing and adverse events as a consequence of treatment in comparison to other modalities, a PRISMA‐compliant systematic review (Systematic Review Registration Number: 204016) of twelve electronic databases was conducted for the terms “ablative laser” and “skin resurfacing” from March 2002 until July 2020. Studies included meta‐analyses, randomized control trials, cohort studies, and case reports to facilitate evaluation of the data. All articles were evaluated for bias. The search strategy produced 34 studies. Of 1093 patients included in the studies of interest, adverse events were reported in a total of 106 patients (9.7%). Higher rates of adverse events were described in nonablative therapies (12.2% ± 2.19%, 31 events) when compared with ablative therapy (8.28% ± 2.46%, 81 events). 147 patients (13.4%) reported no side effects, 68 (6.22%) reported expected, transient self‐resolving events, and five (0.046%) presented with hypertrophic scarring. Excluding transient events, ablative lasers had fewer complications overall when compared with nonablative lasers (2.56% ± 2.19% vs 7.48% ± 3.29%). This systematic review suggests ablative laser use for skin resurfacing is a safe and effective modality to treat a range of pathologies from photodamage and acne scars to hidradenitis suppurativa and posttraumatic scarring from basal cell carcinoma excision. Further studies are needed, but these results suggest that ablative lasers are a superior, safe, and effective modality to treat damaged skin.     

New fillers for the new man

ABSTRACT:  Two new collagen-based lidocaine-containing dermal fillers, ArteSense™/ArteFill™ (Artes Medical, San Diego, CA) and Evolence® (Colbar LifeScience Ltd., Herzliya, Israel), have proved to be of particular interest to men, many of whom seek a long-lasting or permanent correction. ArteFill™ has been available in the United States since 2006, and it is expected that Evolence® will reach the American market in 2008. The properties of the two products will be described, and experience based on the administration of many hundreds of syringes of both products by a Canadian dermatologist will be detailed here, with tips and precautions to optimize patient outcomes.     

Genetic alterations in RAS‐regulated pathway in acral lentiginous melanoma

Studies integrating clinicopathological and genetic features have revealed distinct patterns of genomic aberrations in Melanoma. Distributions of BRAF or NRAS mutations and gains of several oncogenes differ among melanoma subgroups, while 9p21 deletions are found in all melanoma subtypes. In the study, status of genes involved in cell cycle progression and apoptosis was evaluated in a panel of 17 frozen primary acral melanomas. NRAS mutations were found in 17% of the tumors. In contrast, BRAF mutations were not found. Gains of AURKA gene (20q13.3) were detected in 37.5% of samples, gains of CCND1 gene (11q13) or TERT gene (5p15.33) in 31.2% and gains of NRAS gene (1p13.2) in 25%. Alterations in 9p21 were identified in 69% of tumors. Gains of 11q13 and 20q13 were mutually exclusive, and 1p13.2 gain was associated with 5p15.33. Our findings showed that alterations in RAS‐related pathways are present in 87.5% of acral lentiginous melanomas.     

Deutliche Einschränkung der Lebensqualität bei Patienten mit Pemphigus vulgaris: Ergebnisse der deutschen Bullous Skin Disease (BSD)‐Studiengruppe

Background: Pemphigus vulgaris is a potentially life‐threatening autoimmune disorder of the skin and mucous membranes characterized by antibodies against epidermal adhesion molecules. Clinically characteristic are painful chronic blisters or erosions of mucous membranes and skin. There are no published studies on the impact o this disease on quality of life. Patients and methods: This registration was performed within the scope of the German BSD (Bullous Skin Disease) study group, from November 1997 until January 2002. A total of 36 patients with the first diagnosis of pemphigus vulgaris were registered at the university hospitals of Dresden, Erlangen, Kiel, Mannheim, München and Würzburg. Thirty of the 36 (83 %) patients participated in the quality of life questionnaire utilizing the German version of ‘Dermatology Life Quality Index’ (DLQI) provided by A. Y. Finlay. The DLQI varies from 0 to 30 with an increased DLQI score indicating a decrease in quality of quality. Results: The overall DLQI total score of 10 ± 6,7 in the investigated pemphigus patients was significantly increased in comparison to other skin diseases. Conclusions: These results suggest that the DLQI can be a very useful additional outcome criteria for clinical studies with pemphigus vulgaris and in the treatment of these patients.     

Mutations in mevalonate pathway genes in patients with familial or sporadic porokeratosis

Porokeratosis comprises heterogeneous keratinization disorders that are characterized by one or more atrophic patches surrounded by a ridge‐like cornoid lamella. In this study, we evaluated seven families affected by porokeratosis and five sporadic patients of the disease in a Chinese population. We performed Sanger sequencing of exons and flanking intron–exon boundaries of mevalonate pathway genes (MVD, MVK, PMVK and FDPS) and of SLC17A9. In five familial and three sporadic patients, we detected six variations, including four novel mutations (MVD c.1A>G; p.Met1?, c.916G>A; p.Ala306Thr, c.1013+1G>A, and PMVK c.65A>G; p.Lys22Arg) and two recurrent mutations (MVD c.746T>C; p.Phe249Ser, and MVK c.1028T>C; p.Leu343Pro). We then applied I‐TASSER and iGEMDOCK to assess these variants for probable functional impacts. The findings of this study extend the mutation spectrum of porokeratosis and provide further evidence for the genetic basis of this disease.     

Occupational allergic contact dermatitis caused by decorative plants

12 cases of occupational allergic contact dermatitis caused by decorative plants were diagnosed in a 14-year period. The patients were middle-aged, and their average exposure time was 13 years. The plant families and plants causing occupational contact dermatitis were Compositae (5 patients: chrysanthemum, elecampane, gerbera, feverfew), Alstroemeriaceae (5 patients, Alstroemeria ), Liliaceae (4 patients; tulip, hyacinth). Amaryllidaceae (2 patients: narcissus) and Caryophyllaceae (2 patients; carnation, cauzeflower). The known chemical allergens causing dermatitis were tuliposide-A and sesquiterepene lactones, such as alantolactones and parthenolide, in the Liliaceae and Compositae families. 7 of the 12 patients were able to continue their work; 5 were not because of severe relapses of skin symptoms. The plant allergen and extract series currently available are of great help in the diagnosis.     

[Translated article] Pruritus in Dermatology: Part 1—General Concepts and Pruritogens

Pruritus is the most common symptom of dermatologic and systemic diseases. The diagnosis of pruritus is clinical, although additional tests may be necessary to identify or confirm the cause. Translational medicine has led to the discovery of new mediators of itch, or pruritogens, as well as new receptors. Knowing how to properly recognize the main pathway that mediates itch in each patient is the key to successful treatment. Although the histaminergic pathway predominates in conditions like urticaria or drug-induced pruritus, it is the nonhistaminergic pathway that predominates in nearly all other skin diseases covered in this review. Part 1 of this 2-part review discusses the classification of pruritus, additional testing, the pathophysiology of itch and the pruritogens implicated (including cytokines and other molecules), and central sensitization to itch.