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Dai X.-Y.,Third Peoples Hospital of Hangzhou | Yan Y.-L.,Zhejiang Chinese Medical University | Wu Q.-F.,Zhejiang Chinese Medical University | Yu C.-H.,Zhejiang Academy of Medical science | And 2 more authors.
Journal of Ethnopharmacology | Year: 2014

Ethnopharmacological relevance Quyu Qingre granules (QYQRGs) are useful traditional Chinese composite prescription in the treatment of blood stasis syndrome. Comparing differences of pharmacokinetic properties of compounds in QYQRG between normal and blood stasis syndrome rabbits can provide much helpful information. The primary objective of this study was to compare the pharmacokinetics of rhein and chrysophanol after orally administering 2.0 g/kg b.w. QYQRG in normal and acute blood stasis model rabbits. Materials and methods The blood samples were collected subsequently at 5, 10, 15, 20, 30, 45, 60, 75, 90, 120, 240, 360 and 480 min after orally administrating QYQRG. The concentrations of rhein and chrysophanol in rabbit plasma were determined by HPLC and main pharmacokinetic parameters were obtained. Results The pharmacokinetic parameters AUC0-∞, Tlag, C max and K21 of both rhein and chrysophanol were markedly different in the acute blood stasis model rabbits. It was also found that parameters A, β, MRT and T1/2β of rhein and the parameters α and T1/2α of chrysophanol all exhibited significant difference between the normal and acute blood stasis model rabbits. Conclusions The absorption time of rhein and chrysophanol was accelerated and the absorption amount of these two compounds was increased in rabbits with acute blood stasis, suggesting that rhein and chrysophanol would possibly be the two effective compounds in QYQRG. © 2014 Elsevier Ireland Ltd.

Wang S.,Third Peoples Hospital of Hangzhou | Liu D.,Third Peoples Hospital of Hangzhou | Ning W.,Third Peoples Hospital of Hangzhou | Ning W.,Zhejiang Chinese Medical University | Xu A.,Third Peoples Hospital of Hangzhou
Experimental Dermatology | Year: 2015

Considerable evidence implicates that viral infection might be a participant factor in the pathogenesis of vitiligo. However, it is still unclear how viral infection leads to the melanocyte destruction. To elucidate the effects of viral dsDNA on the viability and cytokine synthesis of normal human melanocytes and to explore the underlying mechanisms, primary cultured normal human melanocytes were transfected with poly(dA:dT). The results demonstrated that poly(dA:dT) triggered apoptosis instead of pyroptosis in melanocytes. Knocking down AIM2 or RIG-I by RNA interference partially reduced the poly(dA:dT)-induced LDH release, suggesting the involvement of both nucleic acid sensors in the process of melanocyte death. Poly(dA:dT) induced the expression of pro-inflammatory cytokine genes including IFN-β, TNF-α, IL-6 and IL-8 as well, whereas the pro-inflammatory cytokine production was suppressed by RIG-I siRNA, but not by AIM2 siRNA. Poly(dA:dT) treatment increased the phosphorylation of p38 and JNK and NFκB. Accordingly, NFκB inhibitor Bay 11-7082 and JNK inhibitor SP600125 blocked the induction of the cytokine genes except IFN-β. The production of IL6 and IL8 was also suppressed by p38 inhibitor SB203580. On the contrary, the Poly(dA:dT)-induced melanocyte death was only decreased by SP600125. This study provides the possible mechanism of melanocyte destruction and immuno-stimulation in vitiligo by innate immune response following viral infection. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Hong W.S.,Third Peoples Hospital of Hangzhou | Hu D.N.,Tissue Culture Center | Qian G.P.,Third Peoples Hospital of Hangzhou | McCormick S.A.,Tissue Culture Center | Xu A.E.,Third Peoples Hospital of Hangzhou
British Journal of Dermatology | Year: 2011

Summary Background Autologous melanocytes can be expanded in vitro, allowing the treatment of large lesions of vitiligo in one session. Theoretically, this procedure could provide a higher donor/recipient size ratio (DR ratio) compared with that in noncultured cell transplantation (with a DR ratio < 1: 10). However, the exact DR ratio obtained from this procedure has not been reported. Objectives To study whether transplantation of cultured pure melanocytes at a high DR ratio is as efficient as that at a low DR ratio. Methods One hundred and two patients with vitiligo were treated by transplantation of cultured pure melanocytes and were divided into two groups: a low DR ratio group, including patients with DR ratio ≤ 1: 10 (mean 1: 8, 35 cases) and a high DR ratio group with DR ratio > 1: 10 (mean 1: 27, 67 cases). The extent of repigmentation between these two groups was compared. Results There was no significant difference in repigmentation between the low DR ratio group (mean ± SD 77·4 ± 22·5%) and the high DR ratio group (77·6 ± 24·8%). Multiple regression analysis showed that even after adjustment for age, sex, type of vitiligo and transplanted cell density, there was no significant correlation between the extent of repigmentation and the DR ratio, indicating that patients treated with high DR ratio obtained a satisfactory result and showed no difference from the low DR ratio group. Conclusions Various surgical procedures for the treatment of vitiligo which involve melanocyte transplantation or skin grafts have different inherent DR ratios. Transplantation of cultured pure melanocytes is an expensive and complicated procedure; however, it provides the highest DR ratio (> 1: 10 and up to 1: 60). Surgeons can select one of these methods for the treatment of vitiligo based on their experience and skill, on the size of lesions, and the availability of laboratory support. © 2011 British Association of Dermatologists.

Hong W.S.,Third Peoples Hospital of Hangzhou | Hu D.N.,Tissue Culture Center | Qian G.P.,Third Peoples Hospital of Hangzhou | McCormick S.A.,Tissue Culture Center | Xu A.E.,Third Peoples Hospital of Hangzhou
Journal of the European Academy of Dermatology and Venereology | Year: 2011

Background Transplantation of autologous cultured pure melanocytes is a well-established procedure for the treatment of refractory and stabilized vitiligo. However, there was no report specifically comparing the efficacy with the regard to defined age groups (children-adolescence-adult). Objective We analysed the efficacy of this procedure in the treatment of vitiligo in children and adolescents and compare it with the results in adults treated during the same period and using identical procedures. Methods Melanocytes were isolated from the roof of suction blister, cultured and expanded with Hu16 medium in vitro, and transplanted to laser-denuded receipt area. A total of 12 children (8-12 years), 20 adolescents (13-17 years) and 70 adults with vitiligo were treated using this procedure. Results The patients obtained satisfactory results (repigmentation of 50% or more) results in children, adolescents and adults were 83.3%, 95.0% and 84.0% respectively. The mean extent of repigmentation in children, adolescents and adults was 80.7%, 78.9% and 76.6% respectively. There was no statistical difference in repigmentation among these three groups. After adjusting for all factors (gender, type of vitiligo, period of stability, location of the lesion or transplanted cell density) individually or totally using multiple regression analysis, age still did not correlate to the extent of repigmentation. Conclusions The satisfactory results obtained in the treatment of vitiligo in children and adolescents by transplantation of cultured autologous pure melanocytes are comparable with the results in adults. Therefore, this procedure can be considered in refractory and stable vitiligo in children and adolescents, especially in patients with large vitiliginous lesions. © 2010 European Academy of Dermatology and Venereology.

Zhu Y.,Zhejiang University | Wang S.,Third Peoples Hospital of Hangzhou | Xu A.,Third Peoples Hospital of Hangzhou
Experimental Dermatology | Year: 2013

The paucity of vitiligo animal models limits the understanding of vitiligo pathogenesis and the development of therapies for the skin disorder. In this study, we developed a new mouse model of vitiligo by topically applying the skin-depigmenting agent monobenzone on mice. We demonstrated that monobenzone-induced skin depigmentation on the non-exposed sites and that the severity of lesions depended on drug dosage. The result of the histological examination of the depigmented skin indicated loss of epidermal melanocytes and perilesional accumulation of CD8+ T cells. Furthermore, the monobenzone-induced depigmentation of the Rag1 gene knockout did not appear on the non-exposed sites, supporting the involvement of infiltrating CD8+ T cells in melanocyte destruction. Resemblance in histological characteristics and pathogenesis between monobenzone-induced depigmentation and active human vitiligo suggests good potential of our mouse model for use in vitiligo research. © 2013 John Wiley & Sons A/S.

Yan Y.,Third Peoples Hospital of Hangzhou | Yuan Y.,Third Peoples Hospital of Hangzhou | Liang L.,Third Peoples Hospital of Hangzhou | Chen T.,Third Peoples Hospital of Hangzhou | And 2 more authors.
Medical Science Monitor | Year: 2014

Background: The aim of this study was to investigate the influence of carotid artery stenting (CAS) on the cognition and quality of life of elderly patients with severe stenosis of the internal carotid artery. Material and Methods: 65 elderly patients with symptomatic severe stenosis of internal carotid artery were recruited into 2 groups: the pharmacotherapy group (n=29) and the CAS group (n=36). Before surgery and 1, 3, 6, and 12 months after surgery, Montreal cognitive assessment (MoCA) was used for the evaluation of cognition and WHOQOL-BREF was used for the assessment of quality of life. Results: At 12 months after surgery, total MoCA score and WHOQOL-BREF score in the pharmacotherapy group was significantly reduced when compared with those before surgery (P<0.05). In the CAS group, the total MoCA score, scores of attention and delayed recall, and WHOQOL-BREF score increased significantly at different time points after surgery when compared with those before surgery (P<0.05). Moreover, in CAS group, the MoCA score and WHOQOL-BREF markedly increased gradually over time (P<0.05). Compared with the pharmacotherapy group, cognition and quality of life in the CAS group were improved dramatically during the follow-up period (P<0.05). Conclusions: Severe stenosis of the internal carotid artery is a cause of cognition impairment, and CAS may improve cognition and quality of life. © Med Sci Monit, 2014.

Qiu J.,Third Peoples Hospital of Hangzhou | Yang G.,Third Peoples Hospital of Hangzhou | Shen Z.,Third Peoples Hospital of Hangzhou | Xie Y.,Third Peoples Hospital of Hangzhou | Wang L.,Third Peoples Hospital of Hangzhou
International Journal of Colorectal Disease | Year: 2013

Purpose: The ability to predict tumor sensitivity toward radiotherapy is important in the personalized application of preoperative radiotherapy for patients with rectal cancer. The aim of the present study was to test the human phosphatidylethanolamine-binding protein 4 (hPEBP4) as a predictive marker of tumor response to preoperative radiotherapy in patients with rectal cancer. Method: A retrospective analysis was conducted, consisting of 86 patients with locally advanced rectal cancer who underwent short-course preoperative radiotherapy (20 Gy in five fractions for 1 week) followed by a radical resection. Both pretreatment biopsy specimens and resected primary tumor tissue were collected. Immunohistochemistry and tumor regression grading system were used to evaluate the expression of hPEBP4 in the pretreatment biopsy specimens and the response of rectal cancer to radiotherapy, respectively. Expression of hPEBP4 was correlated with tumor regression in the resected specimen and the clinical outcome of the patients as well. Results: We found that high expression of hPEBP4 was associated with radioresistance in both univariate and multivariate analyses, and patients with a high hPEBP4 expression had a poorer progression-free survival than those with low hPEBP4 expression. Conclusion: Our study revealed the independent predictive values of hPEBP4 in response of rectal cancer to preoperative radiotherapy, which suggests that upregulating hPEBP4 might be a potential mechanism by which rectal cancer cells avoid the destructive effects of radiotherapy. © 2012 Springer-Verlag.

Wu J.,Third Peoples Hospital of Hangzhou | Zhou M.,Third Peoples Hospital of Hangzhou | Wan Y.,Providence College | Xu A.,Third Peoples Hospital of Hangzhou
Molecular Medicine Reports | Year: 2013

Cell-mediated autoimmunity has been suggested to be involved in the melanocyte apoptosis that occurs in vitiligo. We investigated the cytotoxicity to autologous melanocytes of CD8+ T cells from the perilesional margins and peripheral blood samples of vitiligo patients. CD8+ T cells isolated from skin biopsied from the edges of depigmented skin patches of vitiligo patients or from peripheral blood samples of the same donors were proliferated in culture medium. The primary cultures of CD8+ T cells and autologous melanocytes were mixed at ratios of 1:1, 1:2 or 1:5 and incubated for 3 days. The apoptosis of the melanocytes was analyzed by flow cytometry. Secreted cytokines in selected samples were measured by cytokine arrays. The results show that the CD8+ T cells were successfully isolated from the vitiligo perilesional margins. This cell population showed a significantly higher percentage of CD69 expression (56.13±3.55 versus 29.93±2.35%, p<0.01) and CD137 expression (41.74±1.06 versus 25.97±1.63%, p<0.01) compared with CD8+ T cells in peripheral blood from the same donors. The co-culturing of CD8+ T cells from lesional skin with autologous melanocytes induced apoptosis in the melanocytes (16.63±1.21, 16.71±0.63 and 18.32±1.60% for CD8+ T cells and autologous melanocytes at ratios of 1:1, 1:2 and 1:5, respectively). IL-6 levels were much higher in the co-culture (3.01-fold higher than in a melanocyte monoculture and 17.32-fold higher than in a CD8 + T-cell monoculture). The CD8+ T cells were also demonstrated to secrete more IL-13. Taken together, our data demonstrate that the infiltration of active CD8+ T cells takes place in the vitiligo perilesional margins. Those CD8+ T cells present significantly higher activation levels and higher cytotoxicity to autologous melanocytes than their counterparts from peripheral blood samples. These data suggest that CD8 + T cells are likely to be involved in the pathogenesis of vitiligo. © 2013 Spandidos Publications Ltd.

Zhang D.-M.,Third Peoples Hospital of Hangzhou | Hong W.-S.,Third Peoples Hospital of Hangzhou | Fu L.-F.,Third Peoples Hospital of Hangzhou | Wei X.-D.,Third Peoples Hospital of Hangzhou | Xu A.-E.,Third Peoples Hospital of Hangzhou
Dermatologic Surgery | Year: 2014

Background Vitiligo is an acquired skin disorder with great social impact. It can be successfully treated using cultured autologous melanocytes transplantation. Objective To evaluate the effect of different modalities of narrow-band ultraviolet B (NB-UVB) therapy on the outcome of cultured autologous melanocyte transplantation in treating vitiligo. Methods Patients undergoing cultured autologous melanocyte transplantation were randomly assigned to four different study groups. Group 1 underwent 20 sessions of NB-UVB treatment before transplantation; Group 2 underwent 30 sessions of NB-UVB treatment after transplantation; Group 3 underwent 20 sessions of NB-UVB treatment before transplantation and 30 sessions after transplantation; Group 4 underwent only transplantation. Results Four hundred thirty-seven patients were enrolled. Group 3 responded best, more than 90% repigmentation was achieved in 81.3% of patients, and 94.8% patients experienced 50% or greater repigmentation. Statistical analysis showed that there was a highly significant difference between the four groups (χ2 = 35.56, p <.001). Homogeneous skin color was obtained on the repigmentation areas, and no scarring or other serious side effects were observed. Conclusions Cultured autologous melanocyte transplantation is an effective treatment for stable vitiligo. Combination of NB-UVB therapy with melanocyte transplantation can accelerate repigmentation of transplanted vitiliginous areas, especially if NB-UVB is given before and after transplantation. © 2014 by the American Society for Dermatologic Surgery, Inc. Published by Wiley Periodicals, Inc.

Lai L.-G.,Third Peoples Hospital of Hangzhou | Xu A.-E.,Third Peoples Hospital of Hangzhou
Skin Research and Technology | Year: 2011

Background/purpose: Hypopigmentary skin disorders such as vitiligo, nevus depigmentosus and nevus anemicus are common diseases in clinic. The lesions of these diseases could be similar to some extent, although each of them has its own characteristic clinical appearance and histological features. Clinically, the atypical lesions are often difficult to be differentiated. In vivo reflectance confocal microscopy (RCM) is a non-invasive, repetitive imaging tool that provides real-time images at a nearly cellular histological resolution. Our aim was to investigate the RCM features of vitiligo, nevus depigmentosus and nevus anemicus. Subjects and Methods: A total of 135 patients with a clinical diagnosis of the aforementioned diseases were included in this study. The RCM images from depigmented skin, border of the white macules, adjacent normal-appearing skin and distant normal skin for all patients at the dermo-epidermal junction (DEJ) level were investigated. Results: In the active phase of vitiligo (AVP), the RCM demonstrated a complete loss of melanin in lesional skin in eight (53; 15.1%) patients. In 45 patients (53; 84.9%) of the AVP, part of the bright dermal papillary rings normally seen at the DEJ level disappeared or part of the rings lost their integrity and the content of melanin decreased obviously. In 20 patients (53; 37.7%) of the AVP, highly refractile inflammatory cells could be seen within the papillary dermis in the lesional and adjacent normal-appearing skin, which may indicate the lesion progresses. In addition, part of the dermal papillary rings showed lack of integrity or their brightness decreased in adjacent normal-appearing skin in all the patients of the AVP. It is important to know that the RCM demonstrated an ill-defined border. In the stable phase of vitiligo (SPV), the RCM demonstrates a complete loss of melanin in lesional skin and a clear border in 31 (41; 75.6%) patients; the content of melanin and dermal papillary rings in adjacent normal-appearing skin show no changes. In 10 (41; 24.4%) patients, the dendritic and highly refractile melanocytes arose in the recovery phase of vitiligo, which may indicate the repigmentation of vitiligo. There are three kinds of repigmentation patterns under RCM: marginal, perifollicular and diffuse. Distant normal skin showed no difference from controls in both the active and the SPV. In all the patients with nevus depigmentosus, the content of melanin decreases obviously but the dermal papillary rings are intact. The dermal papillary rings show no differences between lesional skin and adjacent normal-appearing skin of nevus anemicus. Conclusion: Considering our results, RCM may be useful to non-invasively discriminate vitiligo, nevus depigmentosus and nevus anemicus in vivo. © 2011 John Wiley & Sons A/S.

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