Yufeng Chen, Song Xue, Jian Shi, Chunfeng He, Qingchuan Zhang
{"title":"EHD2, a novel HIF target gene, is a promising biomarker in clear cell renal cell carcinoma.","authors":"Yufeng Chen, Song Xue, Jian Shi, Chunfeng He, Qingchuan Zhang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objectives: </strong>The aim of the present study was to determine the clinical value of a novel hypoxia-inducible factor (HIF) target EH domain-containing protein 2 (EHD2) for predicting the outcome of patients with clear cell renal cell carcinoma (ccRCC).</p><p><strong>Materials and methods: </strong>GEPIA public database was searched to determine a possible association between HIF2Α and EHD protein family members, and kidney renal clear cell carcinoma data were used to find the expression profile of EHD proteins in ccRCC samples. A tissue microarray from 70 ccRCC samples was used for immunohistochemical analysis to determine the specific expression pattern of EHD2 in ccRCC samples. In addition, univariate and multivariate analyses were performed to assess the utility of EHD2 as an independent prognostic factor for ccRCC.</p><p><strong>Results: </strong>EHD protein family members were all found to be significantly correlated with HIF2Α expression in ccRCC. However, EHD2 was the only protein that was observed to be overexpressed in ccRCC cancer tissues compared with normal tissues. EHD2 and HIF2Α mRNA expression levels were found to be higher in cancer tissues compared with those in adjacent normal tissue according to reverse transcription-quantitative PCR analysis. Among the 70 patients with ccRCC, EHD2 was overexpressed in 52.8% (37/70). Subsequently, EHD2 was found to be significantly associated with both overall survival (P=0.016) and disease-free survival (P=0.029). Furthermore, by multivariate analysis, EHD2 was an independent prognostic factor for patients with ccRCC.</p><p><strong>Conclusion: </strong>EHD2 is a novel HIF target, based on a relatively large sample of EHD2 research in patients with ccRCC. Furthermore, our study provided evidence that EHD2 can serve as a promising biomarker for predicting ccRCC outcome.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 11","pages":"344-351"},"PeriodicalIF":1.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10695750/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138498362","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}
Zhangfu Li, Jiangbei Yuan, Zilong Yan, Xu Liu, Jikui Liu
{"title":"Long noncoding RNA LINC00665 is a diagnostic biomarker that enhances cell proliferation and migration in hepatocellular carcinoma.","authors":"Zhangfu Li, Jiangbei Yuan, Zilong Yan, Xu Liu, Jikui Liu","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>This study aimed to evaluate the relationship between LINC00665 expression levels and the risk of hepatocellular carcinoma (HCC) in Chinese Han nationality patients and to explore the influence of LINC00665 dysregulation on the proliferation and migration potential of HCC cells.</p><p><strong>Patients and methods: </strong>We investigated the expression of LINC00665 in The Cancer Genome Atlas (TCGA) database. Then, we confirmed the expression of LINC00665 in 54 pairs of surgical tissues from HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction. Furthermore, we manipulated the expression level of LINC00665 and examined the cell proliferation and migration abilities of HCC cells.</p><p><strong>Results: </strong>In the TCGA cohort, a high level of LINC00665 in patients with HCC was significantly associated with tumor stage, tumor differentiation grade, and overall survival. In our HCC patient cohort, overexpression of LINC00665 in patients showed positive correlations with alpha-fetoprotein level, Barcelona Clinic Liver Cancer stage, and tumor differentiation grade. In addition, LINC00665 was upregulated in HCC cells, especially in cells with rapid growth rates and high migration abilities. A new LINC00665 isoform with a length of 1,371 nucleotides was identified in MHCC-97H cells. Interfering with LINC00665 expression weakened the proliferation and migration abilities of HCC cells. In contrast, LINC00665 overexpression enhanced proliferation and migration abilities.</p><p><strong>Conclusion: </strong>LINC00665 was upregulated in HCC tissues and cells and might be used to predict a poor prognosis of HCC patients. In addition, LINC00665 may promote the malignant progression of HCC by enhancing proliferation and migration capacities.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 11","pages":"332-343"},"PeriodicalIF":1.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10695748/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138498364","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}
Jianghui Yang, Zhangtao Shao, Duomi Zhang, Kai Wang
{"title":"Pathogenesis and treatment progress in age-related hearing loss: a literature review.","authors":"Jianghui Yang, Zhangtao Shao, Duomi Zhang, Kai Wang","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Age-related hearing loss (ARHL) is a progressive sensorineural hearing loss caused by age. The pathogenesis of ARHL is not completely clear at present, but it is closely related to auditory nerve degeneration, metabolic disorders, vitamin deficiency, and genetics, especially mitochondrial DNA damage. With the acceleration of industrialization and urbanization in our country, the impact of environmental noise is increasing, and ARHL has become one of the most important factors affecting the quality of life of the elderly in our country. Therefore, hearing intervention for patients with ARHL plays a crucial role in improving their quality of life. At present, the use of hearing aids and cochlear implants are the main means to treat the daily hearing difficulties and communication difficulties of patients with ARHL. However, in China, due to the economy, the concept of not wanting to treat the elderly, and other reasons, the hearing aid wearing rate compared to developed countries has significant differences. Cochlear implant is another option for patients with presbyacusis, and patients can obtain good hearing and speech recognition rate after surgery. At present, there is no definitive conclusion on whether the quality of life of patients after cochlear implantation has been improved, and this study will be reviewed based on previous relevant reports.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 11","pages":"315-320"},"PeriodicalIF":1.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10695747/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138498365","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":"Lenvatinib plus Immune Checkpoint Inhibitors versus Lenvatinib monotherapy as treatment for advanced hepatocellular carcinoma: a meta-analysis.","authors":"Xinlin Yu, Chun Wei, Ran Cui, Ou Jiang","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Lenvatinib, an FDA-approved first-line oral multi-kinase inhibitor for advanced hepatocellular carcinoma (aHCC), has demonstrated promise for treatment. Nevertheless, findings from the Leap-002 study suggest that the addition of anti-vascular drugs to Lenvatinib may not yield significant improvements in survival rate. This meta-analysis aims to comprehensively assess the effectiveness of Lenvatinib, both as a standalone treatment and in combination with immune checkpoint inhibitors (ICIs), in managing advanced aHCC patients. We retrieved relevant studies published up to March 1, 2023, from databases such as PubMed, the Cochrane Library, Web of Science, and Embase. Subsequently, we conducted an analysis using REVMAN 5.3 and Stata MP 14.0 software, following quality assessment and data extraction procedures. A random effects model was employed to calculate the risk ratio (HR) using a 95% confidence interval (CI). The initial literature search yielded 921 results. However, after multiple rounds of exclusion and the removal of unrelated studies, 26 papers met the screening criteria. After a thorough examination of the full texts, we found that 8 studies met the analysis criteria. The combination of Lenvatinib with ICIs demonstrated significant improvement in overall survival (OS) (HR=1.53, 95% CI: 1.34-1.74; P<0.001) and progression-free survival (PFS) (HR=1.51, 95% CI: 1.34-1.72; P<0.001). Furthermore, subgroup analysis, categorized by the duration of follow-up, revealed that for the 3-year combined OS (HR=2.21, 95% CI: 1.79-2.73; Z=7.40, P<0.05), the combination therapy significantly outperformed monotherapy, leading to a 2.21-fold increase in OS for patients during the 3-year follow-up period. Nevertheless, for non-3-year combinations (HR=1.206, 95% CI: 1.020-1.425; Z=2.19, P<0.05), there was merely a 1.206-fold increase in effectiveness compared to single therapy for follow-ups of both longer and shorter durations. This might be attributed to the insufficient representation of HBV-related aHCC cases and the Asian population in the study, along with the increased availability of second-line treatment options for advanced cancer, which can influence the observed effectiveness of immunotherapy.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 11","pages":"321-331"},"PeriodicalIF":1.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10695749/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138498363","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}
Jiejing Yin, Dina Zenezan, Khanh Duy Doan, Alisa Nobee, Shuanzeng Wei, Mehri Mollaee, Daniela M Proca
{"title":"Radiologic and clinicopathologic features of eosinophilic solid and cystic renal cell carcinoma: report of two cases and review of literature.","authors":"Jiejing Yin, Dina Zenezan, Khanh Duy Doan, Alisa Nobee, Shuanzeng Wei, Mehri Mollaee, Daniela M Proca","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Eosinophilic Solid and Cystic Renal Cell Carcinoma (ESC RCC) is a rare entity described in the latest WHO Classification of Urinary and Male Genital Tumours (2022 edition). It is a neoplasm that occurs most often in a sporadic setting, with no association with tuberous sclerosis complex (TSC). It typically presents as a well demarcated, non-encapsulated lesion, with solid and cystic architecture, composed of cells with voluminous eosinophilic cytoplasm and cytoplasmic stippling. Tumor cells are at least focally immunohistochemically (IHC) reactive for CK20. CD10 and Cathepsin K are positive in most cases. Consistent somatic mutually exclusive mutations in the TSC1 and TSC2 genes are detected in ESC RCC. We describe two ESC RCC cases diagnosed at our institution. Both cases occurred in female patients, ages of 33 and 64, respectively. Both patients had no evidence of TSC and both lesions were found incidentally, by imaging studies, at an early stage. Macroscopic and microscopic findings in both neoplasms were classic. One case was analyzed by molecular testing and TSC2 gene mutation was detected. Both cases had focal positivity of CD10 and Cathepsin K by IHC. Both tumors were stage pT1a at diagnosis and the patients remained free of disease after resection. It has been proposed that TSC1/2 can be a molecular marker for ESC RCC and be used to expand the morphologic spectrum of ESC RCC. As a novel rare subtype of renal cell carcinoma, with very limited data on molecular evaluation, it is useful to document these newly diagnosed ESC RCC cases.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"303-308"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641369/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648868","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":"Angiotensin-converting enzyme - human amniotic mesenchymal stem cells improve pulmonary vascular remodeling in rats with pulmonary hypertension by promoting angiogenesis and counteracting inflammation.","authors":"Changfang Wu, Hao Liu, Mingchuan Ba, Jie Zha, Zhen Gao, Lijun Li, Peiyuan Xu, Minfei Li, Fusheng Cai, Mingjie Chen, Xiaona Wu, Lin Guo, Hongzhe Zhang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objectives: </strong>Human Amniotic Mesenchymal Stem Cells (hAMSCs) have strong multidirectional differentiation ability. Studies have found that transfection of target genes into target cells by lentivirus can enhance the differentiation potential of the cells. Angiotensin-Converting Enzyme 2 (ACE2) was found to improve vascular remodeling. Research is lacking on ACE2-hAMSCs. Therefore, this study aimed to investigate the ability to improve pulmonary arterial hypertension using ACE2-hAMSCs.</p><p><strong>Methods: </strong>Lentiviruses overexpressing ACE2 were mixed with hAMSCs. Then, ACE2-hAMSCs and hAMSCs with good growth in logarithmic growth phase were collected. We detected their migration and angiogenesis. RT-qPCR technology was used to detect the expression levels of genes related to angiogenesis, and inflammation in the two cell lines, and western-blotting was used to detect the expression levels of ACE2. As an animal study, 21 rats were randomly divided into four different groups. Right heart hypertrophy, pulmonary angiogenesis, and serum inflammatory factors were measured before dissection. H&E staining was used to observe the inflammatory infiltration of lung tissues.</p><p><strong>Results: </strong>The migration and angiogenesis of ACE2-hAMSCs were strongerthan that of hAMSCs alone. The expressions of genes in ACE2-hAMSCs were higher, and the expression of ACE2 protein in ACE2-hAMSCs was less. H&E staining showed that the inflammatory infilration of lung tissue in ACE2-hAMSCs groups was significantly improved. In addition, the ACE2-hAMSCs group had stronger pro-angiogenesis and anti-inflammatory effects.</p><p><strong>Conclusion: </strong>These results suggest that ACE2-hAMSCs can repair pulmonary vascular endothelial cell injury caused by pulmonary hypertension by promoting angiogenesis and anti-inflammatory ability. This shows that ACE2-hAMSCs have stronger ability to improve pulmonary vascular remodeling than hAMSCs alone.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"282-293"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641367/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648923","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":"Adult cerebral high-grade glioneuronal tumor with perivascular or pseudopapillary growth co-existing with low-grade tumor: a case report.","authors":"Masayuki Shintaku, Makoto Ohta, Hideo Chihara, Hideaki Yokoo, Yuri Noda, Koji Tsuta","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>An unusual, small cell-predominant, high-grade glioneuronal tumor in the occipital lobe of a 49-year-old man that co-existed with a low-grade tumor is reported. The tumor consisted of two distinct components: the major component was a dense proliferation of primitive small cells showing bidirectional neuronal and glial differentiation; and the minor component consisted of a proliferation of well-differentiated astrocytes intermingled with mature neuronal cells. In the former component, perivascular pseudorosette-like or pseudopapillary growth reminiscent of ependymoma or papillary glioneuronal tumor (PGNT), respectively, was prominent, and hypertrophic astrocytic cells were located just outside the central blood vessels. Small cells were immunoreactive for Olig2, synaptophysin, and, less frequently, for glial fibrillary acidic protein. The low-grade component included Rosenthal fibers, hemosiderin deposition, and perivascular lymphocytic infiltration, thus closely resembling ganglioglioma. Cytogenetic studies did not demonstrate any mutations or rearrangements of the genes <i>IDH1, IDH2, H3F3A, BRAF, FGFR1,</i> or <i>TERT</i> promoter. The tumor recurred and spread along the ventricular surface three years after total removal. The small cell-predominant, high-grade component was considered to have evolved from the ganglioglioma-like, low-grade component. The histopathologic resemblance of the high-grade component to PGNT was a special feature.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"294-302"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641370/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134650847","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":"Extrarenal Wilms tumor of the recto-vaginal septum with <i>BRCA2</i> gene mutation: a case report.","authors":"Qijun Chen, Kaixuan Yang, Xiao Tang","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Extrarenal Wilms tumor (ERWT) is rare, and its occurrence in the adult recto-vaginal septum is even more uncommon. Importantly, instances of a BRCA2 gene mutation associated with ERWT have not been documented. In this report, we present an unusual case of ERWT situated in the recto-vaginal septum of a 49-year-old woman, accompanied by a concurrent <i>BRCA2</i> gene mutation. After the tumor's second recurrence, the patient experienced symptomatic relief after administering poly (ADP-ribose) polymerase (PARP) inhibitor therapy. Given the limited exposure and understanding of optimal treatment strategies for this distinct tumor, there is a definite need to accumulate further clinical experiences and insight. Consequently, we propose that genetic testing be considered in cases involving tumor recurrence or metastasis, since this may offer valuable information for identifying targets for therapeutic intervention.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"309-313"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641372/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648924","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":"Immune-related gene prognostic index (IRGPI) for lung adenocarcinoma predicts patient prognosis and immunotherapy response.","authors":"Zheng Zhu, Wei Feng, Xiao-Yan Tan, Pin-Chao Gu, Wei Song, Hai-Tao Ma","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>We searched for a predictive biomarker that also predicts whether patients would benefit from immune checkpoint blockade (ICB) treatment from a few angles, because existing biomarkers no longer wholly replicate the interconnections of distinctive elements in the tumor microenvironment (TME).</p><p><strong>Methods: </strong>We identified 55 pivotal IRGs by performing a WGCNA and univariate Cox regression analysis on a lung adenocarcinoma dataset from the TCGA database. The IRGPI model was then constructed using multivariate Cox regression analysis, which identified 16 genes and verified the use of the GSE68465 database. The AUC of the IRGPI was compared to those of the current biomarkers to determine its predictive potential. Then we examined the molecular and immunological properties of ICB and assessed its effectiveness using CTLA4 expression and TIDE.</p><p><strong>Results: </strong>Patients with a high IRGPI had a later clinical stage, more severe symptoms, and a worse prognosis. Patients with a low IRGPI had a higher immune escape potential and were less responsive to immunotherapy.</p><p><strong>Conclusion: </strong>The IRGPI may be a biomarker for determining the prognosis of patients and whether they respond favorably to ICB therapy.</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"260-281"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641371/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648925","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":"Protective effect of mesenchymal stem cell-conditioned medium on hepatic cell apoptosis after acute liver injury [Retraction].","authors":"","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>[This retracts the article on p. 831 in vol. 6, PMID: 23638214.].</p>","PeriodicalId":13943,"journal":{"name":"International journal of clinical and experimental pathology","volume":"16 10","pages":"314"},"PeriodicalIF":1.4,"publicationDate":"2023-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641368/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648926","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}