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Objective:To investigate clinical efficacy and safety of cultured autologous melanocyte transplantation for the treatment of non-segmental vitiligo accompanied by autoimmune thyroid diseases.Methods:From May 2008 to December 2018, a total of 2 284 patients with non-segmental vitiligo were retrospectively collected, who received cultured autologous melanocyte transplantation in Hangzhou Third People′s Hospital. Among these patients, 75 were also diagnosed with autoimmune thyroid diseases, including hyperthyroidism (42 cases) , hypothyroidism (18 cases) and Hashimoto′s thyroiditis (15 cases) . Efficacy and safety were compared between the vitiligo patients with autoimmune thyroid diseases (concomitant group) and those without (non-concomitant group) . Chi-square test was used to compare enumeration data.Results:Among the 2 284 patients, 1 085 were males and 1 199 were females, with an age of 25.0 ± 1.2 years and a disease duration of 5.1 ± 2.3 years. Six months after transplantation, 1 873 out of 2 209 patients in the non-concomitant group achieved favorable clinical response, with a response rate of 84.8%, including 1 162 achieving complete clinical response (52.6%) ; 46 out of 75 patients in the concomitant group achieved favorable clinical response, with a response rate of 61.3%, including 20 achieving complete clinical response (26.7%) ; the response rate and recovery rate were both significantly lower in the concomitant group than in the non-concomitant group ( χ2 = 29.72, 19.54, respectively, both P < 0.001) . Moreover, the response rate was significantly lower in the hypothyroidism group than in the hyperthyroidism group ( χ2 = 6.61, P = 0.010) . The incidence of isomorphic response at the donor site was significantly higher in the concomitant group than in the non-concomitant group (9.3% vs. 4.3%, χ2 = 4.31, P = 0.038) , so were the recurrence rates of vitiliginous patches at the recipient site after 1, 3, 5 and 10 years (concomitant group: 6.7%, 14.7%, 17.3%, 8.7%, respectively; non-concomitant group: 0.7%, 1.4%, 2.1%, 3.6%, respectively; χ2 = 29.96, 70.69, 67.23, 41.61, respectively, all P < 0.001) . Conclusion:Concomitant autoimmune thyroid diseases negatively affect the efficacy of cultured autologous melanocyte transplantation in the treatment of vitiligo, so effective measures should be taken to prevent isomorphic response and recurrence at the recipient site for patients with non-segmental vitiligo complicated by autoimmune thyroid diseases.
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Objective:To study the characteristics of intestinal microflora in patients with vitiligo, and to analyze the relationship between the changes of intestinal microflora and the incidence of vitiligo, so as to provide new ideas for clinical treatment.Methods:Fecal specimens from 30 patients with vitiligo and 30 healthy adults were collected and analyzed qualitatively by Roche/45 high-throughput sequencing platform. At the same time, macrogenomics was used to analyze the feces of 5 patients with vitiligo and 5 healthy adults to identify the potential regulatory pathways.Results:The bacterial species in the feces of patients with vitiligo were similar to those of healthy people, but the intestinal microbial diversity of patients with vitiligo was significantly reduced ( P<0.01); the abundance of Proteus and Clostridium was significantly reduced at phylum level; at genus level, 7 of them were Bacteroides, Escherichia coli Shigella, Rochella, Bacillus anthracis, Clostridium clostridium, Jordani bacteria. The abundance of RF9 and Prunella-7 decreased significantly ( P<0.01), while the abundance of 4 genera (Rumen Coccus-1, Rumen Coccus UCG, Trichomonas and Streptococcus) increased significantly ( P<0.01). The expression of Streptococcus and Phase Anthraceae in vitiligo patients was significantly different: the former increased by 10.8 times, the latter decreased by 6.517 times, and an intestinal microorganism based on 11 vitiligo-related genera was constructed. The random forest model of bacterial flora showed that AUC of the discriminant model was 0.89 in ROC, and macrogenomic analysis showed that the disorders of vitiligo-related bacterial flora were mainly related to immune-related pathways (such as WNT pathway, Notch pathway), energy metabolism, mitochondrial function and amino acid metabolism (such as phenylalanine metabolism). Conclusions:The diversity of bacterial community in intestinal microecological environment of vitiligo patients is significantly different from those in normal people. The imbalance of bacterial community may be involved in the pathogenesis and development of vitiligo. Supplementation of probiotics may be beneficial to the treatment of vitiligo.
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Objective:To preliminarily evaluate the effect of Fam114A1 on the biological function of melanocytes.Methods:A375 human melanoma cells was used to construct stably Fam114A1-overexpressing or -inhibited cell line by lentiviral transfection, namely overexpression group and expression inhibition group respectively, and A375 cells transfected with an empty lentivirus served as control group. Real-time fluorescence-based quantitative PCR was performed to evaluate effect of Fam114A1 on the mRNA expression of melanin synthesis-related genes tyrosinase (TYR) , tyrosinase-related protein 1 (TYRP1) , premelanosome protein (PMEL) , microphthalmia-associated transcription factor (MITF) and dopachrome isomerase (DCT) , Western blot analysis was conducted to determine the protein expression of TYR and MITF, methyl thiazol tetrazolium (MTT) assay, Transwell migration and adhesion assays were conducted to assess the effect of Fam114A1 on cellular proliferative activity, migratory and adhesive ability of A375 cells respectively. Statistical analysis was carried out by using one-way analysis of variance and Dunnett- t test. Results:Fluorescence microscopy showed that lentivirus-based transfection efficiency was about 90% in the 3 groups. Compared with the control group (0.850 ± 0.120) , the protein expression of Fam114A1 significantly increased in the overexpression group (1.507 ± 0.170, t = 5.888, P = 0.001) , but significantly decreased in the expression inhibition group (0.397 ± 0.120, t = 4.065, P = 0.007) , suggesting that the stably Fam114A1-overexpressing or -inhibited A375 cell line was successfully constructed. Real-time fluorescence-based quantitative PCR and Western blot analysis showed that the mRNA and protein expression of TYR and MITF were significantly lower in the expression inhibition group than in the control group (all P < 0.01) , but did not differ between the overexpression group and control group (all P > 0.05) . Compared with the control group, the expression inhibition group showed significantly increased cellular proliferative activity and adhesive ability ( P = 0.009, 0.001, respectively) , but significantly decreased migratory ability ( P = 0.005) , while the overexpression group only showed significantly increased migratory ability ( P = 0.021) . Conclusions:Fam114A1 can affect the proliferative activity, migratory and adhesive abilities of A375 cells, and the expression of melanin synthesis-related proteins TYR and MITF in A375 cells. Fam114A1 may be a functional protein involved in regulating the biological activity of melanocytes.
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Objective To assess the clinical features and diagnostic index of progressive macular hypomelanosis(PMH).Methods Eight patients with PMH were recruited into this study.Wood's lamp and in vivo confocal laser scanning microscopy(CLSM)were utilized to observe the lesions of all patients.Microbiological culture of lesion specimens from 2 patients was performed.Tissue specimens from 4 patients underwent immunohistochemieal staining with anti-S-100 and anti-TRP-1 antibodies for the detection of melanocyte quantity.Electron microscopy wag utilized to observe ultrastructural features of lesions.Primary culture of melanocytes was carried out with lesional epidermis.Resnits Under Wood's lamp.the lesions of PMH showed punctiform red fluorescence.CLSM revealed complete pigmented tings in lesions with decreased melanin granules compamd with those surrounding normal skin.Microbiological culture grew red fluorescence-producing,gram-positive bacillus which was identified as Propionibacterium acnes.Immunohistochemistry showed no significant difference in the number of S-100-postive cells or TRP-1-positive cells per high power field (× 400)between lesions and surrounding normal skin (8.25±0.96 vs 8.75±1.71,4.25±0.96 vs 4.50±1.29,both P>0.05).Ultrastructural studies showed a large reduction in the number of type Ⅳ melanosomes in lesions of PMH,along with numemus membrane bound bodies and clusteredly distributed,small type Ⅱ-Ⅳ melanosomes.Melanocytes,with morphological similarity to normal melanocytes,were successfully isolated from the lesional tissue,cuhured and identified.Conclusion A primary diagnostic criteria is pro-posed for PMH according to the clinical and experimental studies.
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Objective To establish the rep-PCR DNA fingerprinting technique for typing K. pneumoniae strains and apply it in the epidemiological investigation.Methods The 39 strains of K. pneumoniae were typed by plasmid profile analysis, then their chromosomal DNA were extracted and purified by NaI-lyses-glass-powder absorption method for rep-PCR. Results Different strains of K.pneumoniae showed different rep-PCR fingerprint patterns. Phylogenetic analysis showed that they could be classified into six types, mainly type 1, type 2, and type 3. Otherwise, plasmid profile analysis suggested that there were four types; mainly type 1 which contained only one plasmid.Conclusions (1) Rep-PCR DNA fingerprinting technique developed in this study is enough for epidemiological studies with its high typeability, strong discrimination, simplicity and rapidness. (2) There were three predominant K. pneumoniae transmitted between patients in Taihe Hospital during the last two years, and there might be serious cross-infection among different types.