Transplantation research最新文献

{"title":"Immunoregulatory properties of rapamycin-conditioned monocyte-derived dendritic cells and their role in transplantation.","authors":"Camila Macedo, Hēth Turquist, Diana Metes, Angus W Thomson","doi":"10.1186/2047-1440-1-16","DOIUrl":"10.1186/2047-1440-1-16","url":null,"abstract":"<p><p> In efforts to minimize the chronic administration of immunosuppression (IS) drugs in transplantation and autoimmune disease, various cell-based tolerogenic therapies, including the use of regulatory or tolerogenic dendritic cells (tolDC) have been developed. These DC-based therapies aim to harness the inherent immunoregulatory potential of these professional antigen-presenting cells. In this short review, we describe both the demonstrated tolerogenic properties, and current limitations of rapamycin-conditioned DC (RAPA-DC). RAPA-DC are generated through inhibition of the integrative kinase mammalian target of rapamycin (mTOR) by the immunosuppressive macrolide rapamycin during propagation of monocyte-derived DC. Consistent with the characteristics of tolDC, murine RAPA-DC display resistance to phenotypic maturation induced by pro-inflammatory stimuli; exhibit the ability to migrate to secondary lymphoid tissue (important for 'cross-presentation' of antigen to T cells), and enrich for naturally-occurring CD4+ regulatory T cells. In rodent models, delivery of recipient-derived RAPA-DC pulsed with donor antigen prior to organ transplantation can prolong allogeneic heart-graft survival indefinitely, especially when combined with a short course of IS. These encouraging data support ongoing efforts to develop RAPA-DC for clinical testing. When compared to murine RAPA-DC however, human RAPA-DC have proven only partially resistant to maturation triggered by pro-inflammatory cytokines, and display heterogeneity in their impact on effector T-cell expansion and function. In total, the evidence suggests the need for more in-depth studies to better understand the mechanisms by which mTOR controls human DC function. These studies may facilitate the development of RAPA-DC therapy alone or together with agents that preserve/enhance their tolerogenic properties as clinical immunoregulatory vectors.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"16"},"PeriodicalIF":0.0,"publicationDate":"2012-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3560974/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Urinary proteomic shotgun approach for identification of potential acute rejection biomarkers in renal transplant recipients.","authors":"Håvard Loftheim,&nbsp;Karsten Midtvedt,&nbsp;Anders Hartmann,&nbsp;Anna V Reisæter,&nbsp;Pål Falck,&nbsp;Hallvard Holdaas,&nbsp;Trond Jenssen,&nbsp;Leon Reubsaet,&nbsp;Anders Asberg","doi":"10.1186/2047-1440-1-9","DOIUrl":"https://doi.org/10.1186/2047-1440-1-9","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>Acute rejection (AR) episodes in renal transplant recipients are suspected when plasma creatinine is elevated and other potential causes out ruled. Graft biopsies are however needed for definite diagnosis. Non-invasive AR-biomarkers is an unmet clinical need. The urinary proteome is an interesting source in the search for such a biomarker in this population.</p><p><strong>Methods: </strong>In this proof of principle study, serial urine samples in the early post transplant phase from 6 patients with biopsy verified acute rejections and 6 age-matched controls without clinical signs of rejection were analyzed by shotgun proteomics.</p><p><strong>Results: </strong>Eleven proteins fulfilled predefined criteria for regulation in association with AR. They presented detectable regulation already several days before clinical suspicion of AR (increased plasma creatinine). The regulated proteins could be grouped by their biological function; proteins related to growth and proteins related to immune response. Growth-related proteins (IGFBP7, Vasorin, EGF and Galectin-3-binding protein) were significantly up-regulated in association with AR (P = 0.03) while proteins related to immune response (MASP2, C3, CD59, Ceruloplasmin, PiGR and CD74) tended to be up-regulated ( P = 0.13).</p><p><strong>Conclusion: </strong>The use of shotgun proteomics provides a robust and sensitive method for identification of potentially predictive urinary biomarkers of AR. Further validation of the current findings is needed to establish their potential clinical role with regards to clinical AR diagnosis.</p><p><strong>Trial registration: </strong>ClinicalTrials.gov number NCT00139009.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"9"},"PeriodicalIF":0.0,"publicationDate":"2012-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31294764","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The pig as a model for translational research: overview of porcine animal models at Jichi Medical University.","authors":"Eiji Kobayashi,&nbsp;Shuji Hishikawa,&nbsp;Takumi Teratani,&nbsp;Alan T Lefor","doi":"10.1186/2047-1440-1-8","DOIUrl":"https://doi.org/10.1186/2047-1440-1-8","url":null,"abstract":"<p><p> To improve the welfare of experimental animals, investigators seek to respect the 3R principle (Replacement, Reduction, and Refinement). Even when large animal studies are essential before moving to clinical trials, it is important to look for ways to reduce the number of experimental animals used. At the Center for the Development of Advanced Medical Technology, we consider 'medical' pigs to be ideal preclinical model systems.We have been using both wild-type and genetically modified pigs. We began using this approach about 10 years ago with a 'total pig system' to model human health and disease for the purposes of both medical skill education and the development of new devices and therapeutic strategies.At our Center, medical students and residents use pigs to gain experience with surgical skills and train for emergency procedures after appropriate simulation training. Senior clinicians have also used these models to advance the development of innovative tools for endo- and laparoscopic procedures. The Center focuses on translational research for organ transplantation and stem cell therapy. Several pig models have been established for liver, intestine, kidney, pancreas, and lung transplantation. Mesenchymal stromal cells have been established in green fluorescent protein- and red fluorescent protein-transgenic pigs and tested to trans-differentiate organogenesis. A program to establish induced pluripotent stem cells in the pig is ongoing at our Center.Here, we review our 10 years of activity in this field. Based on our experience in surgical education and research, experimental pigs are valuable models in translational research.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"8"},"PeriodicalIF":0.0,"publicationDate":"2012-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31296463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Treatment of bone loss in patients with chronic liver disease awaiting liver transplantation.","authors":"Daniel Kaemmerer,&nbsp;Benjamin Schmidt,&nbsp;Gabriele Lehmann,&nbsp;Gunter Wolf,&nbsp;Utz Settmacher,&nbsp;Merten Hommann","doi":"10.1186/2047-1440-1-7","DOIUrl":"https://doi.org/10.1186/2047-1440-1-7","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>Most of the patients awaiting liver transplantation already have osteopenia or even osteoporosis by end-stage liver disease.In a retrospective study, we investigated the effect of pre-treatment with oral monthly ibandronate (150 mg), vitamin D3 (800 IU/day) and calcium (1 g/day) for osteopenia and osteoporosis caused by end-stage liver disease in patients before and after liver transplantation (LT).</p><p><strong>Methods: </strong>The bone mineral density (BMD) of the lumbar spine (LS) and the femoral neck was measured prospectively pre- and post-LT in 31 patients with existing pre-transplant osteopenia. Patients had osteopenia of the LS prior to LT (T-score -1.8 ± 1.5) so that the treatment medication was initiated immediately after the diagnosis.</p><p><strong>Results: </strong>The study group showed a permanently increased BMD with significant differences (g/cm²) from baseline up to 12 months post LT at the lumbar spine (LS: pre-LT 0.80 ± 0.11 g/cm², three months: 0.90 ± 0.08 (P <0.005); six months: 0.95 ± 0.11 (P < 0.008); 12 months: 1.00 ± 0.09 -0.85 (P <0.012).</p><p><strong>Conclusion: </strong>The combined pre- and post-operative treatment with oral ibandronate had significantly improved bone mineral density of the lumbar spine at 3, 6 and 12 months post LT. The immediate post-operative bone loss after LT can be significantly avoided by pre-treatment of liver transplant candidates affected by osteopenia.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"7"},"PeriodicalIF":0.0,"publicationDate":"2012-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A simplified subnormothermic machine perfusion system restores ischemically damaged liver grafts in a rat model of orthotopic liver transplantation.","authors":"Tim A Berendsen,&nbsp;Bote G Bruinsma,&nbsp;Jungwoo Lee,&nbsp;Vincent D'Andrea,&nbsp;Qiang Liu,&nbsp;Maria-Louisa Izamis,&nbsp;Korkut Uygun,&nbsp;Martin L Yarmush","doi":"10.1186/2047-1440-1-6","DOIUrl":"https://doi.org/10.1186/2047-1440-1-6","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>Liver donor shortages stimulate the development of strategies that incorporate damaged organs into the donor pool. Herein we present a simplified machine perfusion system without the need for oxygen carriers or temperature control, which we validated in a model of orthotopic liver transplantation.</p><p><strong>Methods: </strong>Rat livers were procured and subnormothermically perfused with supplemented Williams E medium for 3 hours, then transplanted into healthy recipients (Fresh-SNMP group). Outcome was compared with static cold stored organs (UW-Control group). In addition, a rat liver model of donation after cardiac death was adapted using a 60-minute warm ischemic period, after which the grafts were either transplanted directly (WI group) or subnormothermically perfused and transplanted (WI-SNMP group).</p><p><strong>Results: </strong>One-month survival was 100% in the Fresh-SNMP and UW-Control groups, 83.3% in the WI-SNMP group and 0% in the WI group. Clinical parameters, postoperative blood work and histology did not differ significantly between survivors.</p><p><strong>Conclusion: </strong>This work demonstrates for the first time in an orthotopic transplantation model that ischemically damaged livers can be regenerated effectively using practical subnormothermic machine perfusion without oxygen carriers.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"6"},"PeriodicalIF":0.0,"publicationDate":"2012-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryo-preserved porcine kidneys are feasible for teaching and training renal biopsy: \"the bento kidney\".","authors":"Kenjiro Konno,&nbsp;Koichi Nakanishi,&nbsp;Shuji Hishikawa,&nbsp;Hozumi Tanaka,&nbsp;Norishige Yoshikawa,&nbsp;Yoshikazu Yasuda,&nbsp;Eiji Kobayashi,&nbsp;Alan Lefor","doi":"10.1186/2047-1440-1-5","DOIUrl":"https://doi.org/10.1186/2047-1440-1-5","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>The use of patients as the primary teaching modality for learning procedures is being questioned. While there have been advancements in the technology used for performing needle biopsies in both native and transplanted kidneys, there has been little advancement in teaching and training tools. We have developed a portable ex-vivo kidney, the Bento Kidney, using cryo-preserved porcine kidneys for teaching this procedure.</p><p><strong>Methods: </strong>The kidney is thawed, perfused by a pump, covered with skin for realistic haptic feedback, and then used with existing biopsy technology to teach the technique.</p><p><strong>Results: </strong>Thirty porcine kidneys were used in this pilot research, and nine were shipped to physicians at a distant facility. Renal biopsy was then performed using a core biopsy needle and ultrasound guidance. There was some leakage of fluid from all kidneys noted. All trainees felt that the model was realistic, and judged at a mean score of 8.7 (SD 0.8) on a scale of 1 (not useful) to 10 (very useful).</p><p><strong>Conclusions: </strong>This feasibility study demonstrates that cryo-preserved porcine kidneys can be successfully used to teach and train renal biopsy techniques, and provides haptic feedback as well as realistic real-time ultrasound images. Further large scale studies are needed to demonstrate value from the educational point of view for nephrology and transplantation.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"5"},"PeriodicalIF":0.0,"publicationDate":"2012-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Organ preservation using a photosynthetic solution.","authors":"Ippei Yamaoka,&nbsp;Takeshi Kikuchi,&nbsp;Tomohiro Arata,&nbsp;Eiji Kobayashi","doi":"10.1186/2047-1440-1-2","DOIUrl":"https://doi.org/10.1186/2047-1440-1-2","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>Organs harvested from a body lapsing into circulatory deficit are exposed to low O2/high CO2, and reach a critical point where original functionality after transplantation is unlikely. The present study evaluates the effect of respiratory assistance using Chlorella photosynthesis on preservation of the rat pancreas from the viewpoint of donation after cardiac death (DCD).</p><p><strong>Methods: </strong>Gas was exchanged through the peritoneum of rats under controlled ventilation with or without Chlorella photosynthetic respiratory assistance. A gas permeable pouch containing Chlorella in solution was placed in the peritoneum and then the space between the pouch and the peritoneum was filled with an emulsified perfluorocarbon gas carrier. Rat DCD pancreases procured 3 h after cardiac arrest were preserved for 30 min in a cold or mildly hypothermic environment or in a mildly hypothermic environment with photosynthetic respiratory support. The pancreases were then heterotopically transplanted into rats with STZ-induced diabetes.</p><p><strong>Results: </strong>Levels of blood oxygen (PaO2) and carbon dioxide (PaCO2) increased and significantly decreased, respectively, in rats with mechanically reduced ventilation and rats given intraperitoneal photosynthetic respiratory support when compared with those without such support. Transplantation with DCD pancreases that had been stored under photosynthetic respiratory support resulted in the survival of all rats, which is impossible to achieve using pancreases that have been maintained statically in cold storage.</p><p><strong>Conclusion: </strong>Respiratory assistance using photosynthesis helps to improve not only blood gas status in the event of respiratory insufficiency, but also graft recovery after pancreas transplantation with a DCD pancreas that has been damaged by prolonged warm ischemia.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"2"},"PeriodicalIF":0.0,"publicationDate":"2012-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31296437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Open new possibilities in Transplantation Research.","authors":"Edward K Geissler,&nbsp;Alan Jardine","doi":"10.1186/2047-1440-1-1","DOIUrl":"https://doi.org/10.1186/2047-1440-1-1","url":null,"abstract":"","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"1"},"PeriodicalIF":0.0,"publicationDate":"2012-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31296464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differential activation of human T cells to allogeneic endothelial cells, epithelial cells and fibroblasts in vitro.","authors":"Dmitry Samsonov,&nbsp;Christopher Geehan,&nbsp;Craig B Woda,&nbsp;David M Briscoe","doi":"10.1186/2047-1440-1-4","DOIUrl":"https://doi.org/10.1186/2047-1440-1-4","url":null,"abstract":"<p><strong>Unlabelled: </strong></p><p><strong>Background: </strong>In the direct pathway, T cells recognize intact donor major histocompatability complexes and allogeneic peptide on the surface of donor antigen presenting cells (APCs). Indirect allorecognition results from the recognition of processed alloantigen by self MHC complexes on self APCs. In this study, we wished to evaluate the relative contribution of different intragraft cells to the alloactivation of nave and memory T cells though the direct and the indirect pathway of allorecognition.</p><p><strong>Methods: </strong>The processing of membrane fragments from IFN-treated single donor endothelial cells (EC), fibroblasts or renal epithelial cells (RPTEC) was evaluated by DiOC labeling of each cell type and flow cytometry following interaction with PBMC. Direct pathway activation of nave CD45RA+ or memory CD45RO+ CD4+ T cells was evaluated following coculture with IFN-treated and MHC class II-expressing EC, fibroblasts or RPTEC. Indirect pathway activation was assessed using CD45RA+ or CD45RO+ CD4+ T cells cocultured with autologous irradiated APCs in the absence or presence of sonicates derived from IFN-treated allogeneic EC, fibroblasts or RPTEC. Activation of T cells was assessed by [3H]thymidine incorporation and by ELISpot assays.</p><p><strong>Results: </strong>We find that CD14+ APCs readily acquire membrane fragments from fibroblasts and RPTEC, but fail to acquire membrane fragments from intact EC. However, APCs process membranes from EC undergoing apoptosis.There was a notable direct pathway alloproliferative response of CD45RO+ CD4+ T cells to IFN-treated EC, but not to fibroblasts or RPTEC. Also, there was a minimal direct pathway response of CD45RA+ CD4+ T cells to all cell types. In contrast, we found that both CD45RA+ and CD45RO+ CD4+ T cells proliferated following coculture with autologous APCs in the presence of sonicates derived from IFN-treated EC, fibroblasts or RPTEC. By ELISpot, we found that these T cells stimulated via the indirect pathway also produced the cytokines IFN, IL-2, IL-4 and IL-5.</p><p><strong>Conclusions: </strong>Recipient APCs may readily process membrane fragments from allogeneic intragraft cells, but not from EC unless they are undergoing apoptosis. This processing is sufficient for indirect pathway alloactivation of both CD45RA+ and CD45RO+ CD4+ T cells. Only graft vascular EC mediate direct pathway reactivation of CD4+ T cells.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"4"},"PeriodicalIF":0.0,"publicationDate":"2012-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31203456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnosis and treatment of coronary artery disease in hemodialysis patients evaluated for transplant.","authors":"Jose Jg De Lima,&nbsp;Luis Henrique W Gowdak,&nbsp;Flavio J de Paula","doi":"10.1186/2047-1440-1-3","DOIUrl":"https://doi.org/10.1186/2047-1440-1-3","url":null,"abstract":"<p><p> We present a review of current strategies for the diagnosis and treatment of coronary artery disease (CAD) in patients with advanced chronic kidney disease who are on the waiting list for transplants, based on data from the literature and originated from a single-center cohort of 1,250 patients with maximum follow-up of 12 years. We discuss the best way to select patients to be tested for CAD, how to choose the more adequate screening test for CAD and cardiovascular disease, how to select patients for invasive treatment studies and how to treat patients with significant CAD. We also suggest new research avenues to be explored to resolve some problems in this area.</p>","PeriodicalId":89864,"journal":{"name":"Transplantation research","volume":"1 1","pages":"3"},"PeriodicalIF":0.0,"publicationDate":"2012-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/2047-1440-1-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31203999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}