Abstract Background : To study the epidemiological characteristics of atypical Kashin-Beck disease cases without characteristic hand lesions such as interphalangeal joint enlargement and brachydactyly and the characteristics of ankle joint lesions. Methods : According to monitoring data, 4 types of regions (including none, mild, moderate and high Kashin-Beck disease endemic areas) in Heilong jiang and Jilin provinces were selected. All local residents over 40 years old were undergone clinical examanation and X-ray images of hands and feet.Kashin-Beck disease was diagnosed in accordance with the "Kashin-Beck disease diagnosis" standard (WS / T 207–2010) for the diagnosis of Kashin-Beck disease. Osteoarthritis was diagnosed in accordance with the American College of Rheumatology 1995 diagnostic criteria. Results : 119 residents over 40 years old were surveyed in a natural village in the non-endemic area. 1,190 residents over 40 years old were surveyed in 38 endemic KBD areas.A total of 710 patients with Kashin-Beck Disease were detected, including 245 patients with grade I, 175 patients with grade II, 25 patients with grade III, and 265 atypical patients. Among all investigated patients , 92.0% (653/710) had ankle joint changes, and it was 80.0% (196/245) in grade I patients and 95.4% (167/175) in grade Ⅱ. Varying degrees of ankle joint changes were found in both grade III and atypical patients. The grade of Kashin-Beck disease was correlated with the degree of ankle joint change ( P <0.001), and the correlation coefficient r s =0.376. Atypical Kashin-Beck disease patients in mild and severe endemic area of Kashin-Beck Disease were younger than those with typical Kashin-Beck disease. Conclusions : There is a correlation between the degree of ankle joint change and the grade of Kashin-Beck disease. The higher the grade of Kashin-Beck disease, the more serious the change of the ankle joint.
With the advent of intensive combination regimens, an increasing number of patients with unresectable pancreatic cancer (UPC) have regained the opportunity for surgery. We investigated the clinical benefits and prognostic factors of conversion surgery (CS) in UPC patients.We retrospectively enrolled patients with UPC who had received CS following first-line systemic treatment in our center between 2014 to 2022. Treatment response, safety of the surgical procedure and clinicopathological data were collected. We analyzed the prognostic factors for postoperative survival among UPC patients who had CS.Sixty-seven patients with UPC were enrolled (53 with locally advanced pancreatic cancer (LAPC) and 14 with metastatic pancreatic cancer (MPC)). The duration of preoperative systemic treatment was 4.17 months for LAPC patients and 6.52 months for MPC patients. All patients experienced a partial response (PR) or had stable disease (SD) preoperatively according to imaging. Tumor resection was unsuccessful in four patients and, finally, R0 resection was obtained in 81% of cases. Downstaging was determined pathologically in 87% of cases; four patients achieved a complete pathological response. Median postoperative-progression-free survival (PO-PFS) was 9.77 months and postoperative overall survival (PO-OS) was 31.2 months. Multivariate logistic regression analyses revealed that the resection margin and postoperative changes in levels of tumor markers were significant prognostic factors for PO-PFS. No factors were associated significantly with PO-OS according to multivariate analyses.CS is a promising strategy for improving the prognosis of UPC patients. The resection margin and postoperative change in levels of tumor markers are the most important prognostic factors for prolonged PFS. Multidisciplinary treatment in high-volume centers is strongly recommended. Prospective studies must be undertaken to resolve the various problems regarding optimal regimens, the duration of treatment, and detailed criteria for CS.
Pancreatic ductal adenocarcinoma (PDAC) has been widely considered as one of the most lethal malignancies with a 5-year overall survival (OS) rate of only 11%.1 Lots of efforts have been devoted into its early diagnosis, treatment strategies, and pathogenesis, etc. PDAC is characterized as excessive desmoplastic tumor with abundant immune cells infiltration, mainly including macrophages, T cells, immature myeloid cells, and neutrophils. Neutrophils are the most abundant white blood cells in blood and have been regarded as a homogeneous group in the past, which belongs to innate immune system and participates in defensing against pathogens. Recently, accumulating evidences determined that neutrophils had an important role in PDAC and accounted for a substantial proportion of tumor-infiltrating immune and inflammatory cells. Jiang et al.2 found that increased neutrophil infiltration was discovered as a central and prominent affected feature, which occurred in the liver, lung, and stomach at the PanIN stage. Importantly, serum leukotriene B4 (LTB4), derived from neutrophils, was validated for the early detection of PDAC. Considering its prognostic role, Ino et al.3 described relationships between prognosis and infiltrating immune cells in a cohort enrolled 212 PDAC patients who received radical surgical resection and found that tumor-infiltrating neutrophils were positively correlated with macrophages and regulatory T cells infiltrations and closely associated with shorter OS and disease-free survival (FS). This indicated that high tumor-infiltrating neutrophils always present boost immunosuppressive microenvironment and contribute to poor survival. Apart from tumor-infiltrating neutrophils, higher circulating neutrophils have also been found a negative association with outcomes. In our previous report, we noted that circulating neutrophil counts, whether 3 days within preoperation or 1 day after operation, were associated with shorter recurrence-FS (RFS) but not with OS, which may hinted that neutrophils exerted a pro-tumor effect in PDAC.4 Tumor-infiltrating neutrophils were often referred as tumor-associated neutrophils (TANs) in most studies. Wang et al.5 identified a pro-tumor subcluster of neutrophils in PDAC and uncovered the pro-tumor mechanisms of TANs in PDAC microenvironment and revealed the association between high glycolytic activity and pro-tumor functions in TANs. Similar to M1/M2 nomenclature of macrophages, recent studies also suggested that TANs could be divided into N1 and N2 categories based on their distinctive phenotypes: antitumorigenic N1 neutrophils and pro-tumorigenic N2 neutrophils. According to current in vivo/vitro studies, the N1/N2 neutrophil polarization may be depended on the special cytokine milieu, mainly including interferon-β (IFN-β) and TGF-β. However, little is known about the prognostic value of tumor-associated N1/N2 neutrophils in PDAC. We preliminarily collected the PDAC tissues after radical surgery from January 2012 to December 2015 in our institute and stained for tumor-associated N1 and N2 neutrophils as refered.6, 7 The typical cell markers of tumor-associated N1 neutrophils were MPO+CD11b+CD206−, and those of tumor-associated N2 neutrophils were MPO+CD11b+CD206+ by three-color immunofluorescence (IF) staining (Figure 1A). Studies have reported that N1 neutrophils may possess powerful antitumor properties through antibody-dependent or direct cytotoxicity8, ROS-mediated coupling9, etc. On the contrary, N2 neutrophils contributed to tumor angiogenesis by secreting vascular endothelial growth factor and matrix metallopeptidase 9, etc. and suppressing CTLs function by arginase.10 In this study, a total of 77 PDAC patients were preliminarily enrolled. Their median age was 65.0 [interquartile range (IQR) 59.5–70.0] years old, and 63.6% of patients (49 of 77) were male. Approximately 25% and 40% of PDAC patients have reached advanced T stages (T3 and T4) and aggressive N stages (N1 and N2) when surgery, respectively. Other clinicopathological characteristics are displayed in Table S1. The median number of tumor-associated N1 and N2 neutrophils were 10.7 (IQR 7.2–25.2) and 21.0 (IQR 13.7–29.0), respectively. The unpaired analysis indicated that the mean number of tumor-associated N2 neutrophils was 23.0 higher than that of tumor-associated N1 neutrophils (16.7, p = 0.002, Figure 1B). Additionally, the discrepancy between the tumor-associated N1 and N2 neutrophils were also observed in the paired analysis (p = 0.011, Figure 1C). According to the optimal cutoffs of tumor-associated N1 and N2 neutrophils, 22 and 18 cases were respectively classified into the high N1 and N2 neutrophil infiltration group. As shown in Table S1, the lower tumor-associated N1 neutrophil infiltration was significantly associated with easier lymph node metastasis (p = 0.012) and higher TNM stage (p = 0.024), but not with other factors. The higher tumor-associated N2 neutrophil infiltration was significantly correlated with distal location (p = 0.039) and easier lymph node metastasis (p = 0.039), similar to the previous study that more aggressive PDAC preferentially recruited more neutrophils. The median OS and RFS of this cohort were 20 and 15 months, respectively. There were 81.6%, 34.1%, and 26.5% OS rates at 1, 3, and 5 years, respectively, while the 1-, 3-, and 5-year RFS rates were 57.9%, 24.9%, and 19.6%, respectively. The Kaplan–Meier survival analyses showed a significant difference in median OS and RFS between the high tumor-associated N1 neutrophil group and the lower group [OS: not reached versus 18 months, Log-rank p < 0.001, HR = 0.153, 95% confidence interval (CI) 0.089–0.262, Figure 1D; RFS: 84 versus 12 months, Log-rank p < 0.001, HR = 0.209, 95% CI 0.125–0.348, Figure 1E]. Accordingly, the median OS (12 versus 32 months, Log-rank p < 0.001, HR = 5.352, 95% CI 2.066–13.870, Figure 1F) and RFS (8.5 versus 22 months, Log-rank p < 0.001, HR = 3.852, 95% CI 1.660–8.941, Figure 1G) of the more markable tumor-associated N2 neutrophil infiltration group were both statistically shorter. Meanwhile, the univariate and multivariate Cox analyses together with conventional clinicopathological variables demonstrated that both tumor-associated N1 and N2 neutrophils were independent prognostic factors for OS (Table S2) and RFS (Table S3) with the opposite HR values. Accumulative evidences showed that neutrophils had complex interaction with tumor microenvironment and played an important role in PDAC progression. The predictive values of neutrophil relevant biomarkers have been found in PDAC patients and regarded as potential therapeutic target. High density of total TANs infiltration always indicated poor outcome in PDAC, and current strategies of anti-TANs mainly focused on depleting TANs entirely or blocking chemokines that functioned as TANs recruitment. Series of preclinical studies used Ly6G or CXCR2 to block TANs and had encouraging outcomes in different mouse models. However, these therapeutic strategies are fraught with difficulties for further clinical trials because of the vital function in defense pathogens of neutrophils. It is worth to consider that TANs have the opposite phenotype between N1 and N2. Our study successfully found that both tumor-associated N1 and N2 neutrophils displayed significant and opposite prognostic values within PDAC. The N1/N2 polarization may depend on different stimuli, including TGF-β induces N2 phenotype whereas IFN-β signaling polarizes neutrophils to N1 phenotype, and the causes of opposite effect in N1 and N2 neutrophil may include the secretions of different cytokines that orchestrate immune cells recruitment, different capability of arginase and proteases synthesis, and direct or indirect cytotoxicity, etc. The aforementioned results hinted that precisely eliminating N2 by specific markers or modified/polarized TANs into N1 would be a more efficacious strategy with less toxicity. In summary, the exploration of neutrophil polarization and its correlation to clinical features in PDAC would significantly enhance our understanding of its pathophysiology and enable us to develop better treatment options. Study conception and design: NP and HY. Acquisition of data: HY, SG, QC, SL, and NP. Analysis and interpretation: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. Drafting of manuscript: NP and HY. Critical revision: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. Statistical analysis: HY, SG, QC, and NP. Study supervision: NP and WW. Read and approved the final manuscript: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. The research was supported by grants from the National Natural Science Foundation of China (grant number: 82103409), China Postdoctoral Science Foundation (grant number: 2021M690037), Shanghai Sailing Program (grant numbers: 21YF1407100 and 21YF1443500), Health Science Popularization Project of Pancreatic Cancer of Zhongshan Hospital (grant number: ZX2021-002), Youth Fund of Zhongshan Hospital Fudan University (grant number: LCBSHZX010), and Shanghai ShenKang Hospital Development Centre Project (grant number: SHDC2020CR2017B). The authors declare no conflict of interest. The study protocol was reviewed and approved by the Ethics Committee of Zhongshan Hospital, Fudan University. Data included in this study are available upon request by contact with the corresponding author. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
AIM:To investigate c-met expression during early pancreatic carcinogenesis. METHODS:We used 46 bulk tissues and 36 microdissected samples, including normal pancreas, chronic pancreatitis, and pancreatic cancer, for quantitative realtime reverse transcription-polymerase chain reaction. RESULTS:In bulk tissue analyses, pancreatic cancer tissues expressed significantly higher levels of c-met than did chronic pancreatitis and normal pancreas tissues.c-met levels did not differ between chronic pancreatitis and normal pancreas tissues.In microdissection-based analyses, c-met was expressed at higher levels in microdissected pancreatic cancer cells and pancreatitisaffected epithelial cells than in normal ductal epithelial cells (both, P < 0.01).Interestingly, pancreatitis-affected epithelial cells expressed levels of c-met similar to those of pancreatic cancer cells.CONCLUSION: Overexpression of c-met occurs during the early stage of pancreatic carcinogenesis, and a single alteration of c-met expression is not sufficient for progression of chronic pancreatitis-affected epithelial cells to pancreatic cancer cells.
Abstract Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy characterized by an immunosuppressive tumor microenvironment enriched with cancer-associated fibroblasts (CAF). This study used a convergence approach to identify tumor cell and CAF interactions through the integration of single-cell data from human tumors with human organoid coculture experiments. Analysis of a comprehensive atlas of PDAC single-cell RNA sequencing data indicated that CAF density is associated with increased inflammation and epithelial–mesenchymal transition (EMT) in epithelial cells. Transfer learning using transcriptional data from patient-derived organoid and CAF cocultures provided in silico validation of CAF induction of inflammatory and EMT epithelial cell states. Further experimental validation in cocultures demonstrated integrin beta 1 (ITGB1) and vascular endothelial factor A (VEGFA) interactions with neuropilin-1 mediating CAF-epithelial cell cross-talk. Together, this study introduces transfer learning from human single-cell data to organoid coculture analyses for experimental validation of discoveries of cell–cell cross-talk and identifies fibroblast-mediated regulation of EMT and inflammation. Significance: Adaptation of transfer learning to relate human single-cell RNA sequencing data to organoid-CAF cocultures facilitates discovery of human pancreatic cancer intercellular interactions and uncovers cross-talk between CAFs and tumor cells through VEGFA and ITGB1.
// Audrey Vincent 1 , Seung-Mo Hong 1 , Chaoxin Hu 1 , Noriyuki Omura 1 , Angela Young 1 , Haeryoung Kim 1 , Jun Yu 1 , Spencer Knight 1 , Michael Ayars 1 , Margaret Griffith 1 , Isabelle Van Seuningen 4,5,6 , Anirban Maitra 1 and Michael Goggins 1,2,3 1 Department of Pathology, the Sol Goldman Pancreatic Cancer Research Center, Johns Hopkins Medical Institutions, Johns Hopkins University, Baltimore, MD, USA 2 Department of Oncology, the Sol Goldman Pancreatic Cancer Research Center, Johns Hopkins Medical Institutions, Johns Hopkins University, Baltimore, MD, USA 3 Department of Medicine, the Sol Goldman Pancreatic Cancer Research Center, Johns Hopkins Medical Institutions, Johns Hopkins University, Baltimore, MD, USA 4 Inserm, UMR837, Jean-Pierre Aubert Research Center, Lille Cedex, France, 5 Université Lille Nord de France, Lille Cedex, France, 6 Centre Hospitalier Régional et Universitaire de Lille, Lille Cedex, France. Correspondence: Michael Goggins, email: // Keywords : EYA2, pancreatic cancer, epigenetic Received : February 14, 2014 Accepted : March 20, 2014 Published : March 22, 2014 Abstract To identify potentially important genes dysregulated in pancreatic cancer, we analyzed genome-wide transcriptional analysis of pancreatic cancers and normal pancreatic duct samples and identified the transcriptional coactivator, EYA2 (Drosophila Eyes Absent Homologue-2) as silenced in the majority of pancreatic cancers. We investigated the role of epigenetic mechanisms of EYA2 gene silencing in pancreatic cancers, performed in vitro and in vivo proliferation and migration assays to assess the effect of EYA2 silencing on tumor cell growth and metastasis formation, and expression analysis to identify genes transcriptionally regulated by EYA2. We found loss of tumoral Eya2 expression in 63% of pancreatic cancers (120/189 cases). Silencing of EYA2 expression in pancreatic cancer cell lines correlated with promoter methylation and histone deacetylation and was reversible with DNA methyltransferase and HDAC inhibitors. EYA2 knockdown in pancreatic cancer cell lines increased cell proliferation. Compared to parental pancreatic cancer cells, pancreatic cancers stably-expressing EYA2 grew more slowly and had fewer metastases in orthotopic models. The transcriptional changes after stable expression of EYA2 in pancreatic cancer cells included induction of genes in the TGFbeta pathway. Epigenetic silencing of EYA2 is a common event in pancreatic cancers and stable expression EYA2 limits the growth and metastases of pancreatic adenocarcinoma.
'/%3e%3cuse xlink:href='%23a' transform='rotate(144 450 300)'/%3e%3cuse xlink:href='%23a' transform='rotate(216 450 300)'/%3e%3cuse xlink:href='%23a' transform='rotate(288 450 300)'/%3e%3c/svg%3e)
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)