Coexistence of Subcutaneous Panniculitis-Like T-Cell Lymphoma and Dermatomyositis in a 12-Year-Old Boy

Subcutaneous panniculitis-like T-cell lymphoma (SPTL) is an extremely rare, indolent skin malignancy that can be difficult to distinguish from autoimmune disease-associated panniculitides. Here, we describe a 12-year-old boy who was diagnosed at age 7 years with dermatomyositis with classical manifestations, including poikiloderma, Gottron’s sign, and symmetric muscle weakness. Recently, the boy presented multiple subcutaneous nodules and fever. Histopathological examination and immunohistochemical staining revealed coexistence of SPTL. To our knowledge, this is the first case of dermatomyositis accompanied with SPTL. This case alert clinical physicians of the possibility of SPTL should be considered when a patient with dermatomyositis has new lesions presenting as nodules and unknown fever.

Rare subtypes of epidermolysis bullosa: three case reports and their pedigree analysis / 中华皮肤科杂志

Objective:To report 3 cases of rare subtypes of hereditary epidermolysis bullosa.Methods:Clinical data were collected from the probands and their relatives, whole-exome sequencing was performed to screen disease-causing mutations in the probands, and Sanger sequencing or qPCR was conducted to verify the mutations in patients and their relatives.Results:Case 1 mainly presented with linear red scars on the back, and the proband, her mother with similar clinical manifestations and her asymptomatic daughter all carried a mutation c.4573G>A (p.Gly1525Arg) in the COL7A1 gene. Case 2 presented with generalized reticular pigmentation all over the body and occasional blisters restricted to the hand and foot, and carried a de novo mutation c.74C>T (p.Pro25Leu) in the KRT5 gene. Case 3 presented with pigmentation abnormalities mainly located at the sun-exposed sites and incomplete syndactyly of the left hand, and carried homozygous deletion mutations in exons 2-6 of the FERMT1 gene, which were inherited from her asymptomatic parents. Case 1 was diagnosed with dominant dystrophic epidermolysis bullosa pruriginosa, case 2 was diagnosed with epidermolysis bullosa simplex with mottled pigmentation, and case 3 was diagnosed with Kindler epidermolysis bullosa. Conclusion:The clinical manifestations of epidermolysis bullosa vary greatly, and gene detection is very important for confirmation of diagnosis of its rare types.

Efficacy and safety of 0.03％ tacrolimus ointment in the long-term intermittent maintenance treatment of atopic dermatitis in children: a multicenter randomized controlled clinical trial / 中华皮肤科杂志

Objective To compare the efficacy and safety of the long-term intermittent maintenance treatment with tacrolimus 0.03％ ointment versus traditional treatment in reducing relapses and prolonging the recurrence interval in children with moderate to severe atopic dermatitis (AD).Methods A two-phase randomized,open-labelled,controlled clinical trial was conducted from September 2012 to November 2013.In the first phase,a total of 171 children aged 2-15 years with moderate to severe AD were enrolled from 7 hospitals in China,and received conventional treatment with tacrolimus 0.03％ ointment twice a day for 2-6 weeks.At the end of the treatment,the patients who achieved an investigator's global assessment (IGA) score ≤ 2 (n =125) were randomly classified into 2 groups to receive the second-phase treatment:test group (n =62) receiving intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week (Monday and Thursday),and control group (n =63) receiving no treatment.If the patients in the 2 groups experienced relapse,they received conventional treatment with tacrolimus 0.03％ ointment twice a day.The overall observation period was 6 months.The primary endpoint was the time to the first relapse,which was defined as the number of days from the end of the first-phase treatment to the first relapse.The secondary endpoints included the number of relapses at the second-phase trial,the disease severity at the time of relapse,the duration of relapse,the pruritus score at the time of relapse,the total amount of tacrolimus ointment used,the total response rate at the second-phase trial,and the incidence of adverse events.Results A total of 125 children with AD were enrolled into the second-phase trial,and 121 of them completed the follow-up.Among the 121 patients,the recurrence rate was significantly lower in the test group (25/60,41.7％) than in the control group (46/61,75.4％;x2 =14.20,P ＜ 0.001).The time to the first relapse was significantly longer in the test group (46.9 ± 37.7 d) than in the control group (28.8 ± 32.3 d;Z =1 093.50,P =0.020).The total number of recurrence was 31 and 86 in the test group and control group respectively,and the mean number of recurrence in each patient was significantly lower in the test group (0.52 ± 0.68) than in the control group (1.41 ± 1.23,t =4.96,P ＜ 0.001).There were no significant differences between the two groups regarding disease severity during relapse (eczema area and severity index:Z =971.50,P =0.39),duration of relapse (Z =747.00,P =0.07),and pruritus score during relapse (Z =894.00,P =0.95).The therapeutic drug was tolerated well in all the children,and no tacrolimus-related serious adverse events occurred.Conclusion The intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week for 6 months can effectively and safely prevent and reduce relapses,and prolong the recurrence interval in children with moderate to severe AD.

Comparative analysis of MRI and clinical characteristics of juvenile dermatomyositis / 临床儿科杂志

Objective To explore the relationship of magnetic resonance imaging (MRI) performance in juvenile dermatomyositis (JDM) with clinical characteristics, especially serum muscle enzyme abnormalities. Methods The images of 50 cases of JDM were reviewed, and the affected skin, subcutaneous connective tissue, muscle fascia and muscle were evaluated. The range of muscle edema under MRI images was categorized into 1-3 grades from mild to complete involvement. By comparing the MRI classification with muscle enzyme, we aim to reveal the linkage between the degree of muscle damage and the levels of muscle enzymes. The sequence of MRI examination of the thigh included rapid spin echo sequence FSE T1W, and the fat suppression sequence STIR in both coronary and axial position. Results In the 50 cases of dermatomyositis, there were 49 cases of muscle edema, 28 cases of myofasciitis, 9 cases of subcutaneous connective tissue inflammation, and 4 cases of thickening skin. Of 49 cases of muscle edema, there are 16 cases in grade 1, 24 in grade 2 and 9 in grade 3. The median value of all muscle enzyme in the grade 2 group was higher than that in the other two groups, and the value of AST was statistically significant, P<0.05. Conclusions The detection rate of JDM using MRI is high, which can help judge the severity and range of involvement. The MRI findings of JDM mainly demonstrated muscle edema, mostly moderate degree, followed by muscular fasciitis.

Association of polymorphisms in the filaggrin gene with the occurrence and clinical phenotypes of atopic dermatitis / 中华皮肤科杂志

Objective To investigate the association of polymorphisms in the filaggrin (FLG)gene with the occurrence and clinical phenotypes of atopic dermatitis (AD).Methods A questionnaire survey was carried out to collect data from 261 patients with AD,including the diagnosis of allergic rhinitis and asthma,and the severity of AD.Mixed food allergen screening test and mixed inhaled allergen screening test were performed in a part of patients,so was the detection of total serum IgE and eosinophil cationic protein (ECP).Among the above AD patients and 276 healthy controls,17 polymorphic sites in exon 3 of the FLG gene,including R444G,T454A,P478S,H519N,D836D,S1482Y,A1805V,R1891Q,1961Q,S2166S,Y2194H,H2330H,D2339N,S2366T,E2398Q,K2444E and E2652D,were genotyped by overlapping PCR and DNA sequencing.Results Binary logistic regression analysis and chi-square test showed no correlations between the 17 polymorphic sites in the FLG gene and the occurrence of AD (all P ＞ 0.05).However,the H519N polymorphic site was associated with AD complicated by asthma (x2 =8.680,P =0.011),and the AA genotype of H519N could increase the risk of asthma in the AD patients (P =0.004,OR =1.061,95％ CI:1.016-1.109).The S2366T and K2444E polymorphic sites were associated with food sensitization in the AD patients (x2 =6.520,6.121,P =0.038,0.047,respectively),and the GG + CG genotype of S2366T (P =0.012,OR =1.396,95％ CI:1.054-1.849)and its G allele (P =0.037,OR =1.350,95％ CI:1.008-1.807) both could increase the risk of food sensitization in the AD patients.Similarly,the AA + GA genotype of K2444E (P =0.013,OR =1.393,95％ CI:1.049-1.850)and its G allele (P =0.028,OR =1.380,95％ CI:1.025-1.857) could increase the risk of food sensitization in the AD patients.Conclusions The FLG polymorphisms may be predisposing factors for some AD-related clinical phenotypes in Chinese Han population.The H519N gene may be associated with AD complicated by asthma,and the S2366T and K2444E genes may be related to food sensitization in AD patients.

Molecular analysis of two pediatric cases with sporadic neurofibromatosis type 1 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report on two children manifesting multiple cafe-au-lait spots suspected as neurofibromatosis type 1, and perform NF1 gene mutation analysis.</p><p><b>METHODS</b>Blood samples were collected from the 2 children, their unaffected parents and 100 normal controls. The entire coding region of the NF1 gene was amplified by PCR and subjected to direct sequencing.</p><p><b>RESULTS</b>In patient 1, a novel frameshift mutation c.1948delT (p.Leu650TyrfsX38) was identified in exon 12 of the NF1 gene. And in patient 2, a previously reported nonsense mutation c.541C>T (p.Gln181X) was revealed in exon 4b. The same mutations were not detected in their unaffected parents or 100 normal controls.</p><p><b>CONCLUSION</b>The two patients were diagnosed with neurofibromatosis type 1 by molecular genetic testing. The pathogenic mutations were c.1948delT and c.541C>T, respectively.</p>

Advance in research on causative genes of xeroderma pigmentosum and related diseases / 中华医学遗传学杂志

Ultraviolet light(UV)-sensitive disorders refer to a group of diseases due to damages to the nucleotide excision repair mechanism which cannot effectively repair DNA damage caused by ultraviolet radiation. The inheritance pattern of such diseases, mainly including xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy, is autosomal recessive and known to involve 13 genes. As proteins encoded by such genes are involved in DNA repair and transcription pathways. There is overlap between the symptoms of such diseases, and their genotype - phenotype correlations are quite complex. To facilitate genetic and prenatal diagnosis for such diseases, a summary of the research progress is provided, which mainly focused on mutation research and genotype - phenotype correlation studies. We also propose a strategy for their genetic diagnosis based on recent findings of our group.

Prevalence of atopic dermatitis, ichthyosis and filaggrin mutations in adolescents in a middle school in Shanghai / 中华皮肤科杂志

Objective To investigate the prevalence and progression process of atopic diseases in adolescents, and to assess their relationship with filaggrin(FLG)mutations. Methods Totally, 334 adolescents aged from 11 to 19 years in a middle school in shanghai were enrolled into this study. A clinical interview was carried out to determine the prevalence of atopic diseases (such as ichthyosis, atopic dermatitis (AD), asthma, rhinitis, etc)in these subjects. Peripheral blood samples were collected from 285 out of the 334 adolescents for screening for common FLG mutations, including 3321delA and K4671X. Five years later, these adolescents were followed up for reevaluation of clinical presentations of atopic diseases. Statistical analysis was carried out by the chi-square test with the SPSS 20.0 software. Results As the baseline survey showed, 19 (5.69%)of the 334 adolescents had AD, 14 (4.19%)had ichthyosis vulgaris, 36(10.78%)had allergic rhinitis, and 4(1.20%)had asthma. FLG mutations were observed in 24(8.42%) of the 285 adolescents. Five years later, 265 adoscents completed the follow-up, and 69 (20.66%)were lost to follow-up. Of the 265 adolescents reevaluated, 13(4.89%)had AD, 15(5.64%)had ichthyosis vulgaris, 27(10.15%)had allergic rhinitis, and 1 (0.38%)had asthma. By the time the second survey was performed, 6 out of the 19 patients initially diagnosed with AD had achieved complete regression, 13 had experienced a marked decrease in SCORing atopic dermatitis (SCORAD)score, and symptoms had disappeared in 9 of the 36 patients initially diagnosed with allergic rhinitis. The frequency of FLG mutations was 10.0%in patients with AD, 55.6%in those with ichthyosis, and 40.0%in those with both AD and ichthyosis, and the development of ichthyosis was associated with FLG mutations(P<0.001). Conclusions The frequency of common FLG mutations was 8.42%in these adolescents. FLG gene may be a semidominant gene associated with ichthyosis vulgaris, and multiple factors influence its expression.

Genetic diagnosis of sporadic neurofibromatosis type 1 with café-au-lait spots as the only presentation in a child / 中华皮肤科杂志

Objective To make a genetic diagnosis of sporadic neurofibromatosis type 1 with café-au-lait spots as the only presentation in a child.Methods Blood samples were collected from an 8-year-old child patient,his parents,and 100 healthy human controls.The mutation of NF1 gene was detected by PCR and direct sequencing.Results No mutation was detected in the NF1 gene of the parents or the healthy controls.There was a de novo nonsense mutation c.3520C ＞ T (p.Q1174X) in the NF1 gene of the patient,which leaded to a premature termination codon.Conclusions The child with café-au-lait spots as the only manifestation is diagnosed with sporadic neurofibromatosis type 1 by genetic testing.The mutation c.3520C ＞ T (p.Q1174X) may be an underlying cause of neurofibromatosis type 1.

Efficacy and safety of tacrolimus 0.03 ％ ointment for the treatment of 2-year-old patients with moderate to severe atopic dermatitis / 中华皮肤科杂志

Objective To evaluate the efficacy and safety of tacrolimus 0.03％ ointment for the treatment of 2-year-old patients with moderate to severe AD.Methods An open-labeled,non-comparative,multi-center study was carried out,which included 59 2-year-old children with moderate to severe AD.All the patients were given topical tacrolimus 0.03％ ointment twice daily for 3 weeks.The evaluation of patients was scheduled at the baseline,1,2,and 3 weeks after the start of treatment.Clinical outcome parameters included the total response rate,eczema area and severity index score (EASI score),the percentage of body surface area (BSA％) affected,physician's global evaluation (PGE),children's dermatology life quality index (CDLQI),visual analog scale (VAS) pruritus score.Safety was assessed based on adverse events reported by patients or observed by the physicians.Results At the end of the treatment,the total response rate was 65.85％ with an EASI score of 4.18,and BSA％ of 16.41％.Of these patients,85.10％ achieved a satisfactory outcome,2.13％ achieved a complete cure,and all achieved an improvement,with no exacerbation observed.The 3-week treatment also resulted in a significant decrease in VAS pruritus score (from 6.80 to 3.21) and CDLQI (frown 7.06 to 2.91).Side effects mainly manifested as temporary burning sensation at the application site,and no severe adverse events associated with tacrolimus were observed.Conclusion Tacrolimus 0.03％ ointment seems safe and effective for the treatment of 2-year-old patients with moderate to severe AD.

Rapid identification and sequence typing of genotype VG Ⅱ of Cryptococcus gattii / 中华临床感染病杂志

Objective To establish a PCR method for rapid identification and sequence typing of VG Ⅱ allele of the intergenic spacer region (IGS) of Cryptococcus gattii.Methods Since IGS1 was of high sequence variation,multiple alignments were conducted by ClustalX 2 in IGS1 of Cryptococcus gattii and Cryptococcus neoformans,and then primer sets specific to genotype VG Ⅱ was designed for PCR analysis.The specificity of the primer pair was detected by amplification of the other genotypes in Cryptococcus gattii,Cryptococcus neoformans,and other pathogenic yeasts.The amplified fragments from VG Ⅱ genotype were sequenced and subtyped.Results Using the PCR analysis developed in this study,all VG Ⅱ genotype strains tested were amplified,whereas no amplification was obtained from other genotypes or yeast species involved herein.Three polymorphic nucleotide sites at 72,79 and 104 bp in the fragment amplified could be used to distinguish sub-genotypes within VG Ⅱ genotype.Conclusions The PCR analysis developed in this study can be used for rapid identification of genotype VG Ⅱ of Cryptococcus gattii.The sequence typing based on the amplified fragment from IGS1 may be performed for screening the highly virulent sub-genotype VGⅡ a.

Identification of Cryptococcus neoformans var.grubii, var.neoformans and Cryptococcus gattii by multiplex PCR / 中华皮肤科杂志

Objective To establish a multiplex PCR targeting the intergenic spacer regions (IGS) for the identification of Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii.Methods Primers were designed by using the software ClustalW2 and Oligo 6 based on the sequence of IGS1 region,which shows high sequence variability in the genome of Cryptococcus neoformans and Cryptococcus gattii.,for the multiplex PCR.Then,the developed multiplex PCR was performed to identify 51 Cryptococcus neoformans strains representing genotypes VNI-VNIV and VNB as well as 41 Cryptococcus gattii strains representing genotypes VGI-VGIV.The identification results were compared with those from common PCR by using primers GPA1A,CLA4D and SODlgattii specific to Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii,respectively,as well as with those from the canavanine-glycine-bromothymol blue (CGB) medium-based culture.Results The developed multiplex PCR successfully identified the 92 Cryptococcus neoformans and Cryptococcus gattii.strains,and yielded negative results from the other tested pathogenic yeasts,which revealed a high specificity of the designed primers.False positive results were observed in the identification of two Cryptococcus gattii strains with GPA1A primer-based PCR,one Cryptococcus gattii strain with CLA4D primer-based PCR,one var.grubii strain and one var.neoformans strain with CGB culture,while no false negative results were observed in the detection of these Cryptococcus strains by any of these methods.Conclusions The developed multiplex PCR in this study can rapidly and accurately identify Cryptococcus neoformans var.grubii,var.neoformans,AD hybrid,and Cryptococcus gattii,with superior performance in comparison with common PCR and CGB medium-based culture.

Species profiles of pathogens from 1366 superficial candidiasis cases in Shanghai / 中华皮肤科杂志

Objective To analyze the composition and distribution of pathogens from 1366 superficial candidiasis cases in Shanghai.Methods Candida species identification was carried out for 1366 adults or children with superficial candidiasis by using CHROMagar Candida plates,API20C AUX system,etc.Pal's agar,Xylose assimilation and the test for growth at 45 ℃ were utilized to differentiate Candida dubliniensis.Newly identified pathogenic Candida species including Candida orthopsilosis,Candida metapsilosis,Candida fermentati,Candida nivariensis and Candida bracarensis were differentiated by molecular biological methods.Finally,the composition and distribution of pathogens in superficial candidiasis cases were statistically analyzed.Results A total of 1366 Candida strains,included 2 Candida orthopsilosis strains and 4 Candida metapsilosis strains,were isolated from these cases.Among these isolates,Candida albicans predominated(79.0％),followed by Candida parapsilosis(9.5％),Candida tropicalis(2.9％)and Candida guilliermondii(1.9％).The composition of Candida species was significantly different between child and adult patients(x2 =196.46,P ＜ 0.01),with the isolation rate of non-albicans Candida species being 14.4％ and 45.8％ respectively in child and adult patients.Conclusions Candida albicans is still the dominant pathogen of superficial candidiasis.Candida orthopsilosis and Candida metapsilosis can cause superficial candidiasis.The isolation rote of non-albicans Candida species is higher in adult patients than in child patients.

A preliminary study on the expression and function of μ-opioid system in the epidermis of patients with atopic dermatitis / 中华皮肤科杂志

Objective To investigate the expression of μ-opioid system in the epidermis of patients with atopic dermatitis and its role in the pathogenesis of atopic dermatitis. Methods Thirty-two mice were equally divided into 4 groups, negative control group, pre-treatment group, naloxone group, and physiological saline group. Ovalbumin was used to sensitize mice in pretreatment group, naloxone group, and physiological saline group for 7 weeks, then, mice in naloxone group and physiological saline group were treated with intracutaneous naloxone or physiological saline solution for 1 week, respectively. Mice were killed in negative control group and pre-treatment group at the end of sensitization, and in naloxone group and physiological saline group after 1-week injection with naloxone or physiological saline, skin tissues were obtained from the back of killed mice and subjected to histological examination with HE staining and quantitative fluorescent PCR for the detection of mRNA expression of μ-opioid receptor (MOR) and its ligand (β-endorphin) in epidermis. The atopic dermatitis severity index of lesions and histological changes were assessed before and after the treatment. Results In comparison with the negative control mice, the epidermal expression level of MOR was signifieantly decreased (t = 2.549, P ＜ 0.05 ) in pre-treatment group, but increased in naloxone group and showed no statistical difference from the negative control group (t = 0.671, P ＞ 0.05). No significant difference was observed in the epidermal β-endorphin mRNA expression between negative control group and pre-treatment group or naloxone group (both P ＞ 0.05 ). The improvement of lesions could be visualized after treatment with naloxone (t = 8.338, P ＜ 0.01 ), which was concordant with the histological changes in naloxone group. Conchusions As an antagonist of MOR, naloxone can restore the expression of epidermal MOR in mice model for atopic dermatitis, and shows a certain efficacy in the treatment of atopic dermatitis, which proves that μ-opioid system is somewhat associated with the pathogenesis of atopic dermatitis.

Autologous peripheral hematopoietic stem cell transplantation in the treatment of adult dermatomyositis: a case report / 中华皮肤科杂志

Objective To investigate the efficacy of autologous peripheral hematopoietic stem cell transplantation in the treatment of adult dermatomyositis. Methods A 21-year-old patient with dermatomyositis received autologous peripheral hematopoietic stem cell transplantation and was followed up for 6 years. Autologous peripheral hematopoietic stem cells were mobilized by recombinant human granulocyte colony stimulating factor (rhG-CSF) before the transplantation, and the conditioning regimens consisted of cyclophosphamide,methylprednisolone and cyclosporin. Rabbit anti-human T lymphocyte immunoglobulin began to be applied on day 3 after retransfer of stem cells. The improvement in symptoms, physical signs and biochemical indicators was observed, and hematopoietic restructuration and immunity resurrection were evaluated after the transplantation. Results After the transplantation, skin eruption greatly improved and gradually subsided. The muscle force of extremities restored from level Ⅳ before transplantation to level Ⅴ. The level of creatine kinase declined sharply after transplantation, but gradually returned to previous level. Leucocyte count began to decrease on the day of retransfer, and returned to the normal level on day 8. Immune function remained normal before and after the transplantation. Conclusion Autologous peripheral hematopoietic stem cell transplantation is an alternative treatment for severe and refractory dermatomyositis.

Triterpenoids from leaves of Ilex cornuta / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of Ilex cornuta.</p><p><b>METHOD</b>The chemical constituents were isolated by column chromatographic methods. And the structures were identified by spectral data.</p><p><b>RESULT</b>Seven compounds were isolated and identified as follows: 2alpha-hydroxy ursolic acid (1), arjunolic acid (2), 23-hydroxy ursolic acid (3), 27-0-p-(Z) -coumaroyl ursolic acid (4), 27-O-p-(E)-coumaroyl ursolic acid (5), asiatic acid (6).</p><p><b>CONCLUSION</b>Compound 1, 2 were obtained from this genus for the first time and 3-6 from this plant for the first time.</p>

A case of anhidrotic ectodermal dysplasia with immunodeficiency / 中华皮肤科杂志

To report a case of 16-month-old boy with anhidrotic ectodermal dysplasia with immunodeficiency who experienced disseminated herpes simplex infection. From 2 months of age, the patient experienced multiple pyrexial episodes of undetermined origin, which responded well to anti-inflammatory agents after undressed. Abnormal sweat with dry skin was noted; therefore, the skin biopsy of right axilla was performed at 7 months of age, and suggested a diagnosis of anhidrotic ectodermal dysplasia. Since 6 months of age, he developed recurrent upper respiratory infections and 2 episodes of pneumonia. Twenty days before, several glossal erosions occurred in the patient, supervened by painful and erosive eruptions and numerous blisters around the mouth and both hands with hyperpyrexia. Four days before, the patient was transferred to the department owing to skin lesion exacerbation. Cutaneous examination showed multiple crested or ulcerated plaques distributed eriorificially (mouth and nasal cavity) on the face. Several irregular, demarcated ulcers were scattered on the buttocks, scrotum and lower limbs, surrounded by grouped and umbilicated vesicles arising on erythema. Both hands were swelling, crusting and painful. Dentition was abnormal, and the patient had only 2 upper conical incisors. Routine investigation revealed that white cell count and C-reactive protein extremely elevated. Immunologic profile showed an abnormal distribution of lymphocyte subsets with decreased CD3+ T cells, CD8+ T cells and NK cells. Serum IgM level was slightly low. IgM antibodies to herpes simplex virus type 1 (HSV-1) were detected by serological testing. Based on the above-mentioned features, a diagnosis of anhidrotic ectodermal dysplasia with immunodeficiency and disseminated herpes simplex infection was confirmed. The patient was resolved favourahly after intravenous ganciclovir and antibiotics for 3 weeks without relapse of skin lesions.

Subgenotyping analysis of pathogenic Cryptococcus gattii isolates from China / 中华皮肤科杂志

Objective To assess the subgenotypes of pathogenic Cryptococcus gattii isolates from China and to elucidate the epidemiological links between these domestic isolates and those from other parts of the world. Methods DNA was extracted from 9 clinical isolates of Ctyptococcus gattii from China. The partially variable regions of the three unlinked loci, namely IGS1, PLB1 and GEF1, were amplified and sequenced, and the bioinformation at these loci was obtained from GenBank for multi-locus sequences alignment and phylogenetic analysis. Results Of these 9 clinical isolates, 8 were genotype VG Ⅰ and mating type α with the same sequences at the tested regions as the reference strain WM276, which was a representative isolate of an independent subgenotype; 1 was of genotype VG Ⅱ and mating type α, which was the first report in China, with the tested sequences consistent with those of the referrence strain R272. Sequencing and phylogenetic analysis of GEF1 gene, which was located at mating type locus, successfully identified the genotypes and mating types of all the Cryptococcus gattii isolates involved here. Conclusions Multi-locus sequence analysis shows that causative Cryptococcus gattii isolates of genotype VG Ⅰ in China carry similar sequences at the tested loci in IGS1, PLB1 and GEF1 genes, to a widely distributed subgenotype in the world, and the sequences of the first VG Ⅱ genotype isolate from China resemble the less virulent subgenotype VG Ⅱ b found in Vancouver islands.

Successful prenatal diagnosis following elimination of maternal cell contamination in a family with recessive dystrophic epidermolysis bullosa / 中华皮肤科杂志

Objective To perform a DNA-based prenatal diagnosis in a family with recessive dys-trophic epidermolysis bullosa, and to develop a strategy to eliminate matemal cell contamination in arnniotic fluid samples. Methods Amniocentesis was carried out at gestation week 16, amniotic fluid culture was used to separate fetal cells from maternal blood cells. Peripheral blood was obtained from the proband, and her parents. Genomic DNA was extracted from peripheral blood and aminotic cells. Subsequently, PCR and direct sequencing were performed to detect pathogenic mutations in the COL7A1 gone. Karyotype analysis was used to confirm paternal information in amniotic fluid. Linkage analysis between micro-satellite markers was performed to confirm the fetal genotype. Resulta Centrifugation showed visible contamination of aminotic cells by blood cells. Direct sequencing revealed that the proband was a carrier of both maternal mutation, R525X in exon 12, and paternal mutation, R2610X in exon 105, while the fetus only carried the maternal mutation, R525X. The second direct sequencing and hapiotype analysis after elimination of mater-nal blood cells by amniotic fluid culture confirmed that the fetus was a carrier of maternal mutation with nor-real phenotype. The pregnancy continued and a clinically unaffected girl was born at gestation week 40.Conclusion The accuracy of DNA-based prenatal diagnosis could be improved by the combination of direct sequencing, amniotic fluid culture, karyotype analysis and linkage analysis, etc.

Multi-locus analysis of AD hybrid strains of Cryptococcus neoformans / 中华临床感染病杂志

Objective To identify the AD hybrid strains and its hybrid types within Cryptococcus neoformans.Methods Difierent hybrid types of AD strains were analyzed by PCR 0f STE20 and MF genes within MAT locus and CIA4 and GPal genes out of MAT locus.The PCR-RFLP analysis of g6341 gene was also performed.Results The mating types of 18 AD strains were precisely identified by PCR of STE20 gene,whereas those of H strain were not identified.CL44 gene was better than the GPal gene in PCR identification of the AD hybrids.In the RFLP analysis of g6341 gene,AD strains were grouped into 2 distinct RFLP patterns based on the mating type on serotype A allele.The mating types of AD strains were not identified by the molecular analyses based on the CL44,GPal and g6341 genes.Conclusion It is necessary to use multi-locus analyses of genes within and out of the MAT locus in precise identification of the AD strains and their hybrid types of Cryptococcus neoformans.