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Objective:To investigate differences in intestinal microbiome between adult patients with psoriasis vulgaris and healthy individuals.Methods:Fecal samples were collected from 22 patients with confirmed psoriasis vulgaris and 23 healthy controls in Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College from September 2017 to February 2018. The total DNA of intestinal flora was extracted and amplified, and the next-generation 16S rRNA gene-targeted sequencing was performed to analyze the diversity and distribution of intestinal flora. Species annotation and classification were performed according to Silva database, and rank sum test was used to analyze species differences among samples at different taxonomic ranks; QIIME software (Version 1.9.1) was used to calculate the number of operational taxonomic units (OTUs) and main indices of α diversity (Chao1 index, Shannon index and Simpson index) , and t test to analyze differences in indices; PCoA analysis was performed to analyze the difference in β diversity, and differences in microbial community composition structure were analyzed between the two groups by using permutational multivariate analysis of variance; rank sum test and linear discriminant analysis effect size (LEfSe) analysis were used to evaluate the species difference. Results:No significant difference in the number of OTUs was observed between the psoriasis group (147.55 ± 57.07) and healthy control group (148.96 ± 50.45, t = 0.088, P = 0.930) . In addition, there were no significant differences in the Shannon index, Chao1 index or Simpson index between the psoriasis group (4.08 ± 0.80, 169.52 ± 63.17, 0.87 ± 0.07, respectively) and healthy control group (4.11 ± 0.94, 175.36 ± 53.59, 0.86 ± 0.90, respectively; t = 0.12, 0.34, 0.27, all P > 0.05) . PCoA analysis showed that the first and second principal components explained 49.8% and 15.62%, respectively, of the total variance between the psoriasis group and healthy control group, and permutational multivariate analysis of variance revealed that the β diversity significantly differed between the two groups ( P = 0.011) . Different microbes between the psoriasis group and healthy control group included Firmicutes, Clostridia, Clostridiales, Erysipelotrichales and Erysipelotrichaceae, whose abundance significantly increased in the psoriasis group, as well as Epsilonproteobacteria, Campylobacterales, Campylobacteraceae, Campylobacter, Bacteroidales and Bacteroidaceae, whose abundance significantly increased in the healthy control group. Conclusion:The intestinal microbiome differs between patients with psoriasis vulgaris and healthy individuals, which may serve as potential biomarkers for psoriasis vulgaris.
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<p><b>OBJECTIVE</b>To detect mutation of adenosine deaminase acting on RNA1 (ADAR1) gene in a pedigree affected with dyschromatosis symmetrical hereditaria (DSH).</p><p><b>METHODS</b>Clinical data and peripheral blood samples of the patients from the pedigree were collected. Potential mutations of the ADAR1 gene were screened among 2 patients, 2 unaffected individual from the pedigree as well as 50 unrelated healthy controls by PCR amplification and direct sequencing.</p><p><b>RESULTS</b>A c.3463C>T (p.R1155W) missense mutation of the ADAR gene was identified in the 2 patients, which was absent in the 2 healthy relatives and 50 unrelated controls. The mutation has been previously identified among 5 Chinese families and was the most common mutation site.</p><p><b>CONCLUSION</b>The c.3463C>T missense mutation of the ADAR gene probably underlies the disease in this pedigree.</p>
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Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.
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Objective To compare the performance of 2％ (w/v) agarose gel,2％ (w/v) Metaphor agarose gel and 10％(w/v) nondenaturating polyacrylamide gel in the PCR-restriction fragment length polymorphism (RFLP) analysis of mycobacterial heat shock protein 65 (hsp65) gene.Methods This study included 8 Mycobacteria strains,including clinical isolates and standard strains of Mycobacteria tuberculosis and Mycobacterium intracellulare.Bacterialsuspension of these strains was prepared with the concentration of bacterial cells varying from 10 to 106per milliliter.PCR was performed to amplify the hsp65 gene with a pair of universal primers followed by the digestion of amplicons with two restriction endonucleases,BstE Ⅱ and Hae Ⅲ.Then,the restriction enzyme-digested fragments were subjected to electrophoresis in 2％ agarose gel,2％ Metaphor agarose gel and 10％ nondenaturating polyacrylamide gel respectively.Results As analysis of variance showed,the three gel electrophoresis methods were statistically different in sensitivity for the RFLP analysis of mycobacterial hsp65 gene (F =36.379,P ＜ 0.01).Least significance difference (LSD) procedure revealed that the 2％ agarose gel-based electrophoresis was less sensitive than the 2％ Metaphor agarose gel-and 10％ non-denaturing polyacrylamide gel-based electrophoresis (both P ＜ 0.01),and no significant differences were observed between the 2％ Metaphor agarose gel-and 10％ non-denaturing polyacrylamide gel-based electrophoresis (P ＞ 0.05).Conclusion The 2％ Metaphor agarose gel-and 10％nondenaturating polyacrylamide gel-based electrophoresis methods appear to be more sensitive than the 2％ agarose gel-based electrophoresis method for the PCR-RFLP analysis of mycobacterial hsp65 gene.
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ObjectiveTo investigate the correlation between the methylation status of HLA class Ⅰ genes(HLA-A, -B and -C) in psoriatic epidermis and disease severity in patients with psoriasis vulgaris. MethodsDNA specimens were obtained from the lesional and nonlesional epidermis of 46 patients with psoriasis vulgaris and from the normal skin of 28 human controls. Methylation specific PCR (MSP) was conducted to detect the methylation status of CpG islands in the promoter region of HLA-A, -B and -C genes. The severity of psoriasis was evaluated by psoriasis area and severity index(PASI) scores. ResultsThe percentage of promoter methylation of HLA-B and HLA-C genes was 4.35％(2/46) and 21.74％(10/46), respectively in nonlesional epidermis, 4.35％ (2/46) and 4.35％ (2/46), respectively in lesional epidermis from these patients. No methylation was observed for the promoter of HLA-A, -B or -C gene in the normal control epidermis or for that of HLA-A gene in the nonlesional or lesional epidermis from the patients. The frequency of HLA-C gene promoter methylation in the nonlesional epidermis was significantly higher than that in the lesional epidermis and control epidermis, but was uncorrelated to the disease severity. No significant difference was observed for the methylation frequency of HLA-A or -B gene promoter among the three groups of specimens. Conclusion Abnormal methylation of HLA-C gene promoter is observed in patients with psoriasis vulgaris.
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Objective To assess the clinical and histopathological features as well as treatment of Wells syndrome.Methods The clinical and pathological findings from 7 patients with Wells syndrome were retrospectively reviewed.Results Lesions were located on both lower extremities in 4 patients,on the back in 1 patient,on the face and trunk in 1 patient,and on the buttocks in 1 patient.Clinical manifestations included cellulitis (n =3),urticaria (n =1 ),annular plaques (n =1 ) and papulonodules (n =2).Histopathological examination of skin biopsies showed an infiltrate of numerous eosinophils with occasional flame figures in the dermis of all the patients.Leucocytoclastic vasculitis was found in 3 cases.No triggering factors were found in any of the 7 cases.The lesions nearly subsided in 3 patients after 2-week treatment with oral small-dosage prednisone and tripterygium glycosides.Conclusions Wells syndrome shows a wide diversity of clinical manifestations with distinct histological features.Systemic glucocorticoids and tripterygium glycosides are effective for the control of this condition.
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Objective To evaluate the safety and efficacy of single and local use of a China-made compound betamethasone injection in the treatment of lichen simplex chronicus. Methods A multi-center,randomized, parallel controlled study was conducted. Patients with lichen simplex chronicus were divided into test and control groups to receive a single dose of intralesional compound betamethasone injection made in China or Schering-Plough Labo N.V. Belgium. Patients were visited for the evaluation of efficacy and safety of the China-made injection at the beginning of the treatment (DO), on week 2 (D14) and 4 (D28) after the initiation of treatment. Results A total of 144 patients were enrolled, among which, 68 in the control group and 71 in the test group completed the trial. FAS analysis on week 4 revealed that the response rate and healing rate were 86.11% and 59.72% in the control group, respectively, 86.11% and 54.17% in the test group, respectively (χ2=0.00,0.45,respectively,both P>0.05).There was no severe adverse event in either group after the treatment, and only mild atrophoderma occurred in one patient in the control group, which was improved spontaneously within several weeks of follow-up. There was no statistically significant difference in the occurrence of side reactions between the two groups (P> 0.05). Conclusion The China-made compound betamethasone injection is effective and safe for the treatment of lichen simplex chronicus.
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Objective To assess the efficacy and safety of various spectrum lasers in the treatment of acne vulgaris. Methods Based on the principles and methods of Cochrane systematic reviews, we searched the Cochrane Library (2009, 6 issues), PubMcd, Embase, China Biomedical Literature Database, China Journal Full Text Database, Chinese Scientific Journals Full Text Database. Retrieval time was up to June, 2009. Randomized controlled trials (RCTs) of lasers for acne vulgaris were included. Results Twelve RCTs totaling 367 patients were included. Because the lack of clinical homogeneity, only descriptive analysis was conducted. Acne lesion counts improved significantly with laser therapy. Adverse effects were limited to transient erythema and edema at treatment sites. Treatment-related pains were well tolerated. Conclusions Current evidence demonstrates that all type lasers in treating acne vulgaris is safe and efficacy. However, higher quality RCT research would be needed to verify the effects and status of lasers on acne vulgaris.
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Objective To investigate the role of AP-1 in the pathogenesis of chronic idiopathic urticaria (CIU). Methods By using immunomagnetic separation technology, peripheral blood basophils were isolated from 10 CIU patients and 10 normal human controls followed by the extraction of nuclear protein from the basophils. TransAMTM AP-1 family kit was used to detect the DNA binding activity changes of AP-1 family transcription factors in basophils, and Western blotting to detect the expression of P-c-jun protein. Results There were some differences in the DNA binding activity of AP-1 family transcription factors in basophils between CIU patients and normal controls. The DNA binding activity of Phospho-c-jun, c-fos, Fos-B, Jun-B and Jun-D factors was increased in CIU patients compared with the controls, and the increase in that of P-c-jun and Jun-D was statistically significant (both P < 0.05). There was an insignificant decrease in the DNA binding activity of Fra-1 factor in the CIU patients compared with the controls (P > 0.05). The P-c-jun (Ser73) protein expression was higher in CIU patients than that in the controls (0.527 ± 0.312 vs. 0.435 ± 0.042, P < 0.05),whereas there was no significant difference in the P-c-jun (Ser63) protein expression level. Conclusion Some changes in DNA binding activity of AP-1 and overexpression of P-c-jun (Ser73) protein in basophils may be involved in the pathogenesis of CIU.
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Objective To test the drug susceptibility of 16 Mycobactena tuberculosis isolates from patients with cutaneous tuberculosis,and to provide a basis for the treatment of this entity.Methods Sixteen strains of mycobacterium were isolated from patients with cutaneous tuberculosis.and they were consistently identified as M.tuberculosis by biochemistry and molecular biology.Absolute concentration method was used to test the susceptibility of these isolates to isoniazid.streptomycin.rifampicin and ethambutol.For two strains resistant to streptomycin,PCR and sequencing were performed to analyse the mutation of rpsL gene.Results Out ofthe 16 strains of M tuberculosis.2 were resistant to streptomycin but sensitive to isoniazid,rifampicin and ethambutol,and 14 were sensitive to isoniazid,streptomycin,rifampicin and ethambutol.Sequencing of the rpsL gene revealed a mutation of AAG to AGG at codon 43 in one streptomycinresistant strain and a substitution of CGC bv CAC at codon 54 in another resistant strain.Conclusions In past five years,the resistance ratio of M tuberculosis was low in patients with cutaneous tuberculosis,and streptomycin resistance predominated in these strains.
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Objective To investigate the mRNA expressions of IFN-gamma receptor and TNF-alpha receptor in peripheral blood mononuclear cells (PBMCs) of patients with psoriasis vulgaris and their role in the pathogenesis of psoriasis. Methods Fifty patients with psoriasis vulgaris (37 cases of active psoriasis and 13 cases of stable psoriasis) and 24 healthy human controls were included in this study. PBMCs were isolated from blood samples obtained from all patients and controls. The mRNA expressions of IFN-gamma receptor and TNF-aipha receptor in PBMCs were detected by RT-PCR. The disease severity in patients was evaluated by psoriasis area and severity index (PASI). Results The mRNA expressions of IFN-gamma receptor and TNF-alpha receptor were observed in the PBMCs of all subjects. The mRNA expression levels of IFN-gamma receptor were 0.72 ± 0.17 in healthy controls, 1.11 ± 0.55 in all patients with psoriasis, 1.13 ±0.57 in patients with active psoriasis and 1.03 ± 0.52 in patients with stable psoriasis, respectively. A signifi-cant increase was observed in the expression levels of IFN-gamma receptor mRNA in all psoriatic patients and in patients with active psoriasis compared with those in healthy controls (both P < 0.05), but there was no significant difference between the healthy controls and patients with stable psoriasis (P ＞ 0.05). The expres-sion levels of TNF-alpha receptor mRNA were 2.05 ± 1.34 in healthy controls, 2.70 ± 3.80 in all psoriatic patients, 2.90 ± 4.40 in patients with active psoriasis, 2.14 ± 1.05 in patients with stable psoriasis, respectively;there was no significant difference between psoriatic patients and healthy controls (P ＞ 0.05). However, no correlation was found between the mRNA expression of IFN-gamma receptor, that of TNF-alpha receptor,and disease severity in psoriatic patients. Conclusions The mRNA expression of IFN-gamma receptor in PBMCs is up-regulated in patients with psoriasis vulgaris, which is unrelated to the activity of psoriasis.
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Objective To assess the association between the amino acid polymorphism (Arg64Gln)within the interferon-γ receptor 2 gene (IFN-γR2) and psoriasis vulgaris in Chinese Hans. Methods Blood samples were collected from 182 patients with psoriasis vulgaris and 114 healthy human controls in Jiangsu and Anhui provinces. The amino acid polymorphism (Arg64Gin) within the IFN- γR2 was examined by PCR-restriction fragment length polymorphism (RFLP) and DNA sequencing. Results No significant difference was observed in the amino acid polymorphism (Arg64GIn) within the IFN-γR2 between the psoriatic patients and healthy controls (P > 0.05 ). There was a significant difference between patients with nail involvement and those without in the frequency of Gln64/Gln64 genotype (57.5% vs 38.1%, X~2= 5.33, P < 0.05),andArg64 (Gln64)allele [19.3% (80.7%)vs30% (70%), X~2=5.03, P < 0.05]. The frequencies of Gln64/Arg64 genotype and Gln64/Gln64 genotype in psoriatic patients with nail involvement significantly differed from those in the controls (29.8% vs 49.1%,X~2 = 5.48, P < 0.05; 57.5% vs 35.1%, X~2= 6.23, P <0.05 ), while no significant difference was found between the psoriatic patients without nail involvement and controls. Moreover, significant difference was noted between patients with prior upper respiratory tract infection (as inducements) and those without in the frequency of Arg64/Arg64 genotype (33.3% vs 15.5%, X~2 =4.94, P < 0.05) and Gln64 (Arg64) allele [51.9% (48.1%) vs 35.2% (64.8%), X~2= 5.46, P < 0.05]. Condusion The amino acid polymorphism (Arg64Gln) within the IFN-γR2 may be associated with the nail involvement and upper respiratory tract infection in patients with psoriasis vulgaris.
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Objective To develop a PCR-RFLP method for the identification of eight mycobacterial species. Methods PCR was performed targeting the gene encoding 65-kDa heat shock protein which was common to all mycobacteria. Two restriction enzymes, BstE Ⅱ and Hae Ⅲ, were used to digest the PCR products, and specific restriction patterns of different mycobacteria were obtained. Results The specific restriction patterns of different mycobacteria were identical to the data previously reported. Conclusion We could differentiate M. avium, M. intracellulare, M. kansasii, M. tuberculosis, M. scrofulaceum, M. marinum, M. fortuitum and M. chelonae in one experiment by PCR-RFLP.
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Objective To study the methylation status of p16 gene in peripheral blood mononuclear cell (PBMC) of patients with plaque psoriasis, and to study its significance in the pathogenesis of the disease. Methods Thirty-four cases of psoriatic patients and 35 healthy controls were measured for the status of promoter methylation in p16 gene by methylation specific PCR (MSP). A region containing CpG site in p16 gene promoter was amplified by MSP. The severity of psoriasis was evaluated by PASI scores. Results The methylation rates of p16 gene in PBMC were significantly higher (P
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Objective To study the corelation of mucosal involvement and corticosteroid dosage in the control of bullous pemphigoid(BP). Methods One hundred and three in-patients with bullous pemphigoid hospitalized during 1988 - 2002 were retrospectively analyzed for their mean corticosteroid dosage for controlling the disease and the duration for complete relieving the skin lesions, as well as the duration to start corticosteroid tapering. All the patients were divided into two groups: group A (37 cases) with mucosal involvement and group B (66 cases) without mucosal involvement. Results The mean corticosteroid dosage used to control the disease in group A was much higher than that in group B (t = 3.488, P 0.05). Conclusion Patients with mucosal involvement require a higher dosage of corticosteroids to control the disease.
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Objective To study the effects of tanshinone on the cultured immortalized human sebocytes and explore the mechanism of action of tanshinone in the treatment of acne. Methods MTT assay was applied to determine the effects of cryptotanshinone and tanshinone Ⅱ? at different concentrations on the proliferation of SZ95 sebocytes in 24 h, 48 h and 72 h after incubation. Lipid contents labeled with Nile red in SZ95 cells were analyzed by flow cytometry technique, and the expression of androgen receptor (AR) mRNA of SZ95 cells were detected by RT-PCR. Results Both cryptotanshinone and tanshinoneⅡ? inhibited the proliferation of SZ95 cells in a dose- and time-dependent mode, with the 50% inhibition concentration (IC50) of 7.473 ?mol/L (in 48 h) and 2.146?mol/L (in 72 h) for cryptotanshinone and 6.021 ?mol/L (in 48 h) and 2.25 ?mol/L (in 72 h) for tanshinone Ⅱ?. Additionally, as compared with the control group, the lipid content of SZ95 cells exposed to tanshinone Ⅱ? at 0.125?mol/L was decreased in 48h (P
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Objective To develop a rapid method with high sensitivity and specificity to detect 4 mycobacterial species(M. tuberculosis, M. avium, M. intracellulare and M. kansasii) which are the most common opportunistic Mycobacteria in AIDS patients. Methods The sensitivities and specificities of PCR were determined with different primer pairs targeting various mycobacterial genes. Multiplex PCR with combination of 4 primer pairs was used to detect the template mixtures of either 1, 2 or 3 mycobacterial DNA. Sensitivities of multiplex PCR were measured. Results Specific DNA fragments of 4 mycobacterial species mentioned above could be detected by PCR and sensitivities ranged from 1 ? 101 ~ 1 ? 102 cells/mL, while the other 17 mycobacterial strains were all PCR-negative. Multiplex PCR could amplify the corresponding 1, 2 or 3 DNA fragments, depending on the number of template DNA added, and sensitivities of multiplex PCR ranged from 1 ? 102 ~ 1 ? 103 cells/mL. Conclusions Multiplex PCR is a rapid, sensitive and specific method for differentiation and detection of Mycobacteria.