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Gemcitabine (GEM) resistance is a major challenge for chemotherapy of pancreatic cancer. Previous studies have reported on the role of long noncoding RNA (lncRNA) in tumorigenesis of pancreatic cancer, however, the involvement of lncRNA in the development of GEM resistance of pancreatic cancer remains unclear. In the present study, we demonstrated that the antisense RNA1 of HIF1α (HIF1A-AS1) was significantly elevated in the GEM-resistant pancreatic cancer cells. Gain- and lost-of-function experiments validated that HIF1A-AS1 promoted GEM resistance of pancreatic cancer cells both in vitro and vivo. We further revealed that HIF1A-AS1 upregulated HIF1α expression and thus promoted glycolysis to enhance GEM resistance of pancreatic cancer cells. Mechanistically, HIF1A-AS1 facilitated the interaction between serine/threonine kinase AKT and Y-box–binding protein 1 (YB1), which promoted phosphorylation of YB1 (pYB1). Meanwhile, HIF1A-AS1 recruited pYB1 to HIF1α mRNA that consequently promoted translation of HIF1α. Furthermore, HIF1α promoted HIF1A-AS1 transcription by directly binding to the HIF1α response element in the promoter area of HIF1A-AS1 to form a positive feedback. Consistently, both HIF1A-AS1 and HIF1α were upregulated in pancreatic cancer tissues and associated with poor overall survival. Together, our results underline a reciprocal loop of HIF1A-AS1 and HIF1α that contributes to GEM resistance of pancreatic cancer and indicate that HIF1A-AS1 might serve as a novel therapeutic target for GEM resistance of pancreatic cancer.Significance:These findings show that a reciprocal feedback of HIF1A-AS1 and HIF1α promotes gemcitabine resistance of pancreatic cancer, which provides an applicable therapeutic target.

Supplementary Methods

Cancer cells are known to undergo metabolic reprogramming, such as glycolysis and glutamine addiction, to sustain rapid proliferation and metastasis. It remains undefined whether long noncoding RNAs (lncRNA) coordinate the metabolic switch in pancreatic cancer. Here we identify a nuclear-enriched antisense lncRNA of glutaminase (GLS-AS) as a critical regulator involved in pancreatic cancer metabolism. GLS-AS was downregulated in pancreatic cancer tissues compared with noncancerous peritumor tissues. Depletion of GLS-AS promoted proliferation and invasion of pancreatic cancer cells both in vitro and in xenograft tumors of nude mice. GLS-AS inhibited GLS expression at the posttranscriptional level via formation of double stranded RNA with GLS pre-mRNA through ADAR/Dicer-dependent RNA interference. GLS-AS expression was transcriptionally downregulated by nutrient stress–induced Myc. Conversely, GLS-AS decreased Myc expression by impairing the GLS-mediated stability of Myc protein. These results imply a reciprocal feedback loop wherein Myc and GLS-AS regulate GLS overexpression during nutrient stress. Ectopic overexpression of GLS-AS inhibited proliferation and invasion of pancreatic cancer cells by repressing the Myc/GLS pathway. Moreover, expression of GLS-AS and GLS was inversely correlated in clinical samples of pancreatic cancer, while low expression of GLS-AS was associated with poor clinical outcomes. Collectively, our study implicates a novel lncRNA-mediated Myc/GLS pathway, which may serve as a metabolic target for pancreatic cancer therapy, and advances our understanding of the coupling role of lncRNA in nutrition stress and tumorigenesis.Significance: These findings show that lncRNA GLS-AS mediates a feedback loop of Myc and GLS, providing a potential therapeutic target for metabolic reprogramming in pancreatic cancer.See related commentary by Mafra and Dias, p. 1302

Supplementary Figures

Supplementary Figure S1. Downregulation of GLS-AS facilitates pancreatic cancer proliferation and invasion. Supplementary Figure S2. Downregulation of GLS-AS apparently increased the GLS expression. Supplementary Figure S3. GLS-AS exert its biological function mainly through GLS. Supplementary Figure S4. The transcriptional activity of GLS promoter is not affected by GLS-AS. Supplementary Figure S5. GLS-AS inhibits GLS expression by ADAR1/Dicer-dependent RNA silencing in PANC-1 cells. Supplementary Figure S6. GLS-AS inhibits GLS expression via ADAR1/Dicer-dependent RNA silencing in BxPC-3 cells. Supplementary Figure S7. GLS-AS inhibits GLS expression by ADAR1/Dicer-dependent RNA silencing in BxPC-3 cells. Supplementary Figure S8. Schematic diagram of RNA pulldown by MS2-GST. Supplementary Figure S9. ADAR1 and Dicer are not significantly varied in pancreatic cancer cells under glucose or glutamine deprivation. Supplementary Figure S10. Downregulation of GLS-AS in pancreatic cancer might attributed to energy stress through Myc-depending regulation. Supplementary Figure S11. GLS and GLS-AS level does not affect Myc mRNA expression in pancreatic cancer cells. Supplementary Figure S12. GLS-AS is conversely correlated with Myc and GLS expression in pancreatic cancer. Supplementary Figure S13. GLS-AS suppresses pancreatic cancer cell growth, invasion and migration in vitro. Supplementary Figure S14. GLS-AS inhibits the progression of the pancreatic cancer in vivo proven with PANC-1 cells. Supplementary Figure S15. GLS-AS inhibits the progression of the pancreatic cancer in vivo proven with BxPC-3 cells.

Supplementary Tables

LncRNAs take extensive effects in immune cells to mediate various autoimmune diseases. However, its potential role in development of SLE remains uncertain. We screened out candidate lncRNA in exosomes from SLE patients and the healthy, and was further validated in mouse model by histological examinations in vivo. Additional morphological verification was exploited to validate its biological functions by construction of plasmids in vitro. H19 in serum exosomes of patients was escalated, and could also promote the proliferation and migration of macrophages. Additionally, H19 mainly recruited miR-145-5p and promotes the expression of downstream targeted protein PAI-1. It further promoted the release of SLE-associated proinflammatory factors via Jak2-STAT3 axis, and in turn facilitated the polarization of M1 macrophages. Our study uncovered a potential mechanism of lncRNA H19 in mediating macrophage polarization and leading to the release of proinflammatory factors, providing a new intervention target for treating SLE.

Research has indicated that hypoxia profoundly contributes to chemoresistance of pancreatic cancer (PC), while the precise mechanism has not been fully elucidated. In this study, we report a hypoxic exosomal circular RNA (circRNA)-mediated mechanism of conferred chemoresistance in PC cells. Gemcitabine (GEM) resistance was enhanced in normoxic PC cells incubated with exosomes derived from hypoxic PC cells. CircRNA microarray displayed that circZNF91 was remarkably increased in hypoxic exosomes of PC cells compared with normoxic exosomes. Overexpression of circZNF91 obviously stimulated chemoresistance in PC cells, while knockdown of circZNF91 retarded the hypoxic exosome-transmitted chemoresistance. Mechanistically, the hypoxic-induced exosomal circZNF91 transmitted into normoxic PC cells could competitively bind to miR-23b-3p, which deprives the inhibition of miR-23b-3p on expression of deacetylase Sirtuin1 (SIRT1). Consequently, the upregulated SIRT1 enhanced deacetylation-dependent stability of HIF-1α protein, leading to glycolysis and GEM chemoresistance of recipient PC cells. In addition, we revealed that the increased circZNF91 in hypoxic exosome was attributed to the transcriptional regulation by HIF-1α. Coincidently, transmission of hypoxic exosomes into subcutaneous xenografts in nude mice obviously facilitated the chemoresistance of transplanted PC tumor, which could be reversed by depletion of circZNF91 or upregulation of miR-23b-3p. Furthermore, clinical data showed that circZNF91 was significantly upregulated in PC tissues and correlated with overexpression of glycolytic enzymes and short overall survival time. Collectively, exosomal circZNF91 can function as a cargo mediating the signal transmission between hypoxic and normoxic tumor cells to promote GEM chemoresistance of PC and may potentially serve as a therapeutic target.

Rationale: Previous studies have reported on the role of extracellular acidity in the metastasis of numerous cancers. However, the involvement of long noncoding RNA (lncRNA) in the extracellular acidity-induced cancer metastasis of pancreatic cancer (PC) remains unclear. Methods: Different expression levels of lncRNAs in PC cells under normal and acidic conditions were compared by RNA sequencing (RNA-seq). The effects of antisense lncRNA of metastasis suppressor 1 (MTSS1-AS) on acidic PC cells were assessed by gain- and loss-of-function experiments. Fluorescence in situ hybridization, RNA immunoprecipitation, RNA pull-down, Western blot, luciferase reporter, and Chromatin immunoprecipitation assays were employed to determine the regulatory mechanisms of MTSS1-AS in the acidity-induced metastasis of PC cells. The expression of MTSS1-AS and associated pathways were compared in PC samples and peritumoral normal tissues. Results: RNA-seq demonstrated that MTSS1-AS was significantly downregulated in pancreatic cells cultured with the acidic medium. The overexpression of MTSS1-AS remarkably inhibited the acidity-promoted metastasis of PC cells by upregulating the expression of its sense gene metastasis suppressor 1 (MTSS1). Mechanistically, MTSS1-AS scaffolded the interaction between E3 ubiquitin-protein ligase STIP1 homology and U-box containing protein 1 (STUB1) and transcription regulator myeloid zinc finger 1 (MZF1), leading to ubiquitination-mediated degradation of MZF1. Further, MZF1 inhibited the expression of MTSS1 by binding to the MTSS1 promoter. Thus, the acidity-reduced MTSS1-AS facilitated the stability of MZF1 and its inhibitory effect on MTSS1 transcription, thereby promoting the metastasis of PC cells under acidic conditions. Moreover, MTSS1-AS was transcriptionally repressed by the binding of MYC proto-oncogene (Myc) with initiator (Inr) elements of the MTSS1-AS promoter. Meanwhile, MTSS1-AS mutually repressed the expression of Myc by impairing the MZF1-mediated transcription activation of Myc, thereby forming a negative feedback loop between MTSS1-AS and Myc in acidic PC cells. In accordance with the experimental results, MTSS1-AS and MTSS1 were downregulated in PC and correlated with poor overall survival. Conclusions: The results implicated that a reciprocal feedback loop between Myc and MTSS1-AS contributed to the extracellular acidity-promoted metastasis of PC, and indicated that MTSS1-AS was a valuable biomarker and therapeutic target for PC.

Poor prognosis in pancreatic cancer (PanCa) is partially due to chemoresistance to gemcitabine (GEM). Glucose metabolism has been revealed to contribute to the therapeutic resistance and pluripotent state of PanCa cells. However, few studies have focused on the effects of GEM on cancer cell metabolism, stemness of tumor cells, and molecular mechanisms that critically influence PanCa treatment. We demonstrate that GEM treatment induces metabolic reprogramming, reducing mitochondrial oxidation and upregulating aerobic glycolysis, and promotes stem-like behaviors in cancer cells. Inhibiting aerobic glycolysis suppresses cancer cell stemness and strengthens GEM’s cytotoxicity. GEM-induced metabolic reprogramming is KRAS dependent, as knockdown of KRAS reverses the metabolic shift. GEM-induced metabolic reprogramming also activates AMP-activated protein kinase (AMPK), which promotes glycolytic flux and cancer stemness. In addition, GEM-induced reactive oxygen species (ROS) activate the KRAS/AMPK pathway. This effect was validated by introducing exogenous hydrogen peroxide (H2O2). Taken together, these findings reveal a counterproductive GEM effect during PanCa treatment. Regulating cellular redox, targeting KRAS/AMPK signaling, or reversing metabolic reprogramming might be effective approaches to eliminate cancer stem cells (CSCs) and enhance chemosensitivity to GEM to improve the prognosis of PanCa patients. Keywords: ROS, KRAS, AMPK, aerobic glycolysis, cancer stem cells, pancreatic cancer, chemotherapy

Rationale: Hypoxia has been proved to contribute to aggressive phenotype of cancers, while functional and regulatory mechanism of long noncoding RNA (lncRNA) in the contribution of hypoxia on pancreatic cancer (PC) tumorigenesis is incompletely understood. The aim of this study was to uncover the regulatory and functional roles for hypoxia-induced lncRNA-MTA2TR (MTA2 transcriptional regulator RNA, AF083120.1) in the regulation of PC tumorigenesis. Methods: A lncRNA microarray confirmed MTA2TR expression in tissues of PC patients. The effects of MTA2TR on proliferation and metastasis of PC cells and xenograft models were determined, and the key mechanisms by which MTA2TR promotes PC were further dissected. Furthermore, the expression and regulation of MTA2TR under hypoxic conditions in PC cells were assessed. We also assessed the correlation between MTA2TR expression and PC patient clinical outcomes. Results: We found that metastasis associated protein 2 (MTA2) transcriptional regulator lncRNA (MTA2TR) was overexpressed in PC patient tissues relative to paired noncancerous tissues. Furthermore, we found that depletion of MTA2TR significantly inhibited PC cell proliferation and invasion both in vitro and in vivo. We further demonstrated that MTA2TR transcriptionally upregulates MTA2 expression by recruiting activating transcription factor 3 (ATF3) to the promoter area of MTA2. Consequentially, MTA2 can stabilize the HIF-1α protein via deacetylation, which further activates HIF-1α transcriptional activity. Interestingly, our results revealed that MTA2TR is transcriptionally regulated by HIF-1α under hypoxic conditions. Our clinical samples further indicated that the overexpression of MTA2TR was correlated with MTA2 upregulation, as well as with reduced overall survival (OS) in PC patients. Conclusions: These results suggest that feedback between MTA2TR and HIF-1α may play a key role in regulating PC tumorigenesis, thus potentially highlighting novel avenues PC treatment.

BACKGROUND Menopausal syndrome (MPS) is common in pre/peri-menopausal breast cancer patient receiving luteinizing-hormone releasing hormone analogue (LHRHa). According to the results of TEXT & SOFT study, it could affect patients' life quality seriously and even caused 8%˜23% treatment break. Cimicifuga racemosa (Remifemin) is previously proved effective for MPS in nature menopause patients. We aim to design a prospective randomized research for investigating the effect of Remifemin on MPS which caused by LHRHa in breast cancer (NCT03339882). METHODS The pre/peri-menopausal patients diagnosed as early breast cancer planning for LHRHa were identified and randomly divided into two groups after surgery and chemotherapy if necessary: the Remifemin group (Remifemin 20mg twice a day for 12 weeks) and the control group. The primary endpoint is Kupperman menopause index (KMI). The estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH) levels in blood were also evaluated. RESULTS From January 2017 to December 2017, totally 85 patients (42 in Remifemin group and 43 in control group) were enrolled in Zhejiang Cancer Hospital and there were no significant differences in baseline characteristics including age, BMI, tumor stage, ER, PR, HER2, KMI and hormone levels between two groups (Table 1). At the time of 4, 8, 12 weeks after treatment, the KMI of Remifenmin group were all significantly lower than control group (Table 2, P0.05).There was no significant difference in endometrial thickness, ovarian cyst and Uterine fibroids except cervical cyst (P=0.02). CONCLUSION The use of Remifemin is oncological safe and reliable in preventing MPS which caused by LHRHa in breast cancer. The clinical characteristics of two groups Remifemin groupControl groupP n=42n=43 N(%)N(%) Age(years) 37.74±6.6037.16±6.280.68BMI (body mass index) 21.90±2.8321.65±3.010.69T stage02(4.76)2 (4.65)0.16 113 (30.95)22 (51.16) 227 (64.29)19 (44.19) N stage021 (50.00)24 (55.82)0.63 110 (23.81)12 (27.91) 26 (14.29)5 (11.63) 35 (11.90)2 (4.65) ERPositive30 (71.43)35 (81.40)0.28 Negative12 (28.57)8 (18.60) PRPositive30 (71.43)32 (74.42)0.76 Negative12 (28.57)11 (25.58) HER2Positive8 (19.05)10 (23.26)0.64 Negative34 (80.95)33 (76.74) Category of LHRHaGoserelin32 (76.19)28 (65.12)0.26 Leuprorelin10 (23.81)15 (34.88) Category of endocrine drugNone10 (23.81)6 (13.95)0.20 SERM5 (11.90)11 (25.58) AI27 (64.29)26 (60.47) The endpoints of two groups Remifemin groupControl groupP n=42n=43 KMI*Base line2.45±1.452.30±1.360.62 4th week3.83±1.868.37±4.19 Citation Format: Yu X, Wang C, Liang C, Xuan H, Yang H. Cimicifuga racemosa extract prevents menopausal syndrome in LHRHa treatment of breast cancer: A prospective randomized research [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-11-11.

Gemcitabine (GEM) resistance is a major challenge for chemotherapy of pancreatic cancer. Previous studies have reported on the role of long noncoding RNA (lncRNA) in tumorigenesis of pancreatic cancer, however, the involvement of lncRNA in the development of GEM resistance of pancreatic cancer remains unclear. In the present study, we demonstrated that the antisense RNA1 of HIF1α (HIF1A-AS1) was significantly elevated in the GEM-resistant pancreatic cancer cells. Gain- and lost-of-function experiments validated that HIF1A-AS1 promoted GEM resistance of pancreatic cancer cells both in vitro and vivo. We further revealed that HIF1A-AS1 upregulated HIF1α expression and thus promoted glycolysis to enhance GEM resistance of pancreatic cancer cells. Mechanistically, HIF1A-AS1 facilitated the interaction between serine/threonine kinase AKT and Y-box–binding protein 1 (YB1), which promoted phosphorylation of YB1 (pYB1). Meanwhile, HIF1A-AS1 recruited pYB1 to HIF1α mRNA that consequently promoted translation of HIF1α. Furthermore, HIF1α promoted HIF1A-AS1 transcription by directly binding to the HIF1α response element in the promoter area of HIF1A-AS1 to form a positive feedback. Consistently, both HIF1A-AS1 and HIF1α were upregulated in pancreatic cancer tissues and associated with poor overall survival. Together, our results underline a reciprocal loop of HIF1A-AS1 and HIF1α that contributes to GEM resistance of pancreatic cancer and indicate that HIF1A-AS1 might serve as a novel therapeutic target for GEM resistance of pancreatic cancer. Significance: These findings show that a reciprocal feedback of HIF1A-AS1 and HIF1α promotes gemcitabine resistance of pancreatic cancer, which provides an applicable therapeutic target.

Metastasis-associated protein 2 (MTA2) is overexpressed in certain malignancies, and plays important roles in tumour metastasis and progression. The present study highlights the function of MTA2 in pancreatic carcinoma through its role as a deacetylator of hypoxia-inducible factor-1α (HIF-1α) and a cotranscriptional factor for E-cadherin expression. We found that overexpression of MTA2 promoted, and knockdown of MTA2 inhibited, the invasion and proliferation of pancreatic carcinoma cells both in vitro and in xenograft models in vivo. We also found that MTA2 is transcriptionally upregulated by HIF-1α through a hypoxia response element (HRE) of the MTA2 promoter in response to hypoxia. Reciprocally, MTA2 deacetylates HIF-1α and enhances its stability through interacting with histone deacetylase 1 (HDAC1). Consequently, HIF-1α recruits MTA2 and HDAC1 to the HRE of the E-cadherin promoter, by which E-cadherin transcription is repressed. In agreement with these experimental results, MTA2 is positively associated with HIF-1α, but inversely correlated with E-cadherin, in pancreatic carcinoma samples. Moreover, data from The Cancer Genome Atlas on 172 pancreatic carcinomas indicate an association between high expression of MTA2 and short overall survival. Taken together, our study identifies MTA2 as a critical hub and potential therapeutic target to inhibit the progression and metastasis of pancreatic carcinoma. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Background To explore the impact of age on all-cause death and cancer-specific death in patients with pancreatic duct adenocarcinoma (PDAC) undergoing surgery. Materials and methods Individuals with PDAC undergoing surgery between 2004 and 2013 (N = 11,138) were retrospectively studied from the Surveillance, Epidemiology, and End Results (SEER) cancer registry database. The impact of age on all-cause death and cancer-specific death was assessed using Cox regression model and competing risk model respectively. Results Multivariate Cox regression analysis indicated that the risks of all-cause death increased with age: hazard ratios (95% confidence interval, 95%CI) were 1.10 (1.04–1.17), 1.31 (1.23–1.38), 1.47 (1.35–1.61) for groups 61–70 years, 71–80 years, and >80 years, respectively, compared with ≤60 years. Multivariate competing risk analysis indicated that the risk of cancer-specific death was similar between patients ≤60 years and 61–70 years (subhazard ratio 0.93; 95% confidence interval 0.87–1.00), but decreased in patients 71–80 years (subhazard ratio 0.84; 95%CI 0.79–0.90) and >80 years (subhazard ratio 0.76; 95%CI 0.68–0.85). Conclusion Age at diagnosis appeared to be an independent predictor of prognosis, with reverse impacts on all-cause death and cancer-specific death.

Although several staging systems have been proposed for pancreatic neuroendocrine tumors (pNETs), the optimal staging system remains unclear. Here, we aimed to assess the application of the newly revised 8th edition American Joint Committee on Cancer (AJCC) staging system for exocrine pancreatic carcinoma (EPC) to pNETs, in comparison with that of other staging systems. We identified pNETs patients from the Surveillance, Epidemiology, and End Results (SEER) database (2004–2014). Overall survival was analyzed using Kaplan–Meier curves with the log‐rank test. The predictive accuracy of each staging system was assessed by the concordance index (c‐index). Cox proportional hazards regression was conducted to calculate the impact of different stages. In total, 2424 patients with pNETs, including 2350 who underwent resection, were identified using SEER data. Patients with different stages were evenly stratified based on the 8th edition AJCC staging system for EPC. Kaplan–Meier curves were well separated in all patients and patients with resection using the 8th edition AJCC staging system for EPC. Moreover, the hazard ratio increased with worsening disease stage. The c‐index of the 8th edition AJCC staging system for EPC was similar to that of the other systems. For pNETs patients, the 8th edition AJCC staging system for EPC exhibits good prognostic discrimination among different stages in both all patients and those with resection.

Background:Immunoglobulin G4-related disease (IgG4-RD) is an autoimmune disease with chronic systemic inflammation and fibrosis. The main feature of the disease was diffuse swell of the affected organs, and the serum IgG4 level was increased. Histopathology of the lesions showed infiltration of IgG4+ plasma cells. However, the pathogenesis of IgG4-RD is still unclear[1].Objectives:To explore the clinical characteristic of lymphocyte subsets of IgG4-related disease patients, and make comparisons with healthy controls.Methods:A total of 31 patients with IgG4-RD who were admitted to the Rheumatic Immunology Department of the Second Hospital of Shanxi Medical University from January 2016 to June 2020 were included. We collected their Clinical and laboratory data, and selected 30 age and sex matched healthy people as the control group. Flow cytometry was used to detect the percentage and absolute number of lymphocyte subsets (T, B, NK, CD4+T, CD8+T) and CD4+T subsets (Th1, Th2, Th17, CD4+CD25+Foxp3+Treg) in peripheral blood of IgG4-RD patients and healthy controls.Results:(1)The percentage of CD4+T cells in peripheral blood of IgG4-RD patients was higher than that of healthy controls [45.00(33.97-51.48) vs. 39.36(33.78-43.30), PTable 1.Comparation of absolute number and percentage of peripheral blood lymphocyte subsets between IgG4-RD patients(n=31) and healthy controls (n=30).cell subsetsIgG4-RD (n=31)HC(n=30)P valueB150.59(120.14-212.38)203.27(152.90-244.27)0.089B%8.74(6.46-11.45)10.03(8.26-13.21)0.059NK261.98(178.82-303.08)290.83(179.93-451.45)0.175NK%13.14(9.92-18.10)16.50(11.24-21.75)0.105CD3+T1357.44(992.00-1844.82)1305.81(978.24-1597.94)0.708 CD3+T%72.62(69.32-76.96)71.62(64.97-75.25)0.135CD8+T436.40(342.71-596.86)513.50(359.73-620.53)0.665CD8+T%24.26(19.48-31.27)26.50(20.67-32.90)0.535CD4+T741.00(562.78-1095.52)664.50(585.52-789.97)0.428CD4+T%45.00(33.97-51.48)39.36(33.78-43.30)0.032Th1162.32(108.11-216.61)144.27(81.52-161.66)0.094Th1%19.00(15.24-25.54)18.46(14.86-24.27)0.644Th27.82(5.35-11.78)8.25(5.32-10.87)0.817Th2%1.00(0.76-1.27)1.24(0.89-1.64)0.399Th177.90(5.20-12.23)5.60(3.12-8.47)0.010Th17%1.13(0.70-1.55)0.77(0.43-1.07)0.026Treg24.45(19.76-44.79)34.55(27.29-46.57)0.076Treg%3.37(2.82-5.65)4.96(4.18-6.34)0.003Th1/Th220.00(13.78-36.36)14.97(10.31-21.58)0.135Th1/Treg5.72(2.92-8.86)3.68(2.53-4.77)0.021Th2/Treg0.27(0.16-0.52)0.22(0.15-0.32)0.199Th17/Treg0.29(0.16-0.46)0.15(0.08-0.23)0.002Conclusion:Th17/Treg immune disorder exists in IgG4-RD patients, and it is related to the disease activity, indicating that Th17/Treg imbalance may be an important pathogenesis of IgG4-RD.References:[1]Kamisawa T, Zen Y, Pillai S, et al. IgG4-related disease[J]. Lancet, 2015, 385(9976): 1460-1471.Disclosure of Interests:None declared

Decreased level of serum calcium was commonly seen in critical illness. Hypocalcemia was significantly more frequent in patients with severe form of acute pancreatitis (AP), and a negative correlation was observed between endotoxemia and serum calcium in AP. AP patients with persistent organ failure (POF) show an extremely high mortality. The association underlying calcium and POF in AP has not been characterized.We conducted a retrospective cohort study of adult patients who presented within 72hours from symptom onset of AP at our center between January 2014 and May 2015. Demographic parameters on admission, organ failure assessment, laboratory data and in-hospital mortality were compared between patients with and without POF. Uni-and multi-variate logistic regression analyses were utilized to evaluated the predictive ability of serum calcium.A total of 128 consecutive AP patients, including 29 with POF, were included. Compared to patients without POF, patients with POF showed a significantly lower value of serum calcium on admission (2.11±0.46 vs. 1.55±0.36mmol/L, P0.001). After multivariate logistic analysis, serum calcium remained an independent risk factor for POF (Hazard ratio 0.21, 95% confident interval: 0.08-0.58; P=0.002). A calcium value of 1.97mmol/L predicted POF with an area under the curve (AUC) of 0.888, a sensitivity with 89.7% and specificity with 74.8%, respectively.Our results indicate that serum calcium on admission is independently associated with POF in AP and may serve as a potential prognostic factor.

Centrosomal protein 55 (CEP55) is a microtubule-bundling protein that participants in cell mitosis. It is overexpressed in several solid tumours and promotes the growth and invasion of cancer cells. However, the role of CEP55 in pancreatic cancer (PANC) remains unclear. Herein, upregulated expression of CEP55 (associated with poor prognosis) was detected in PANC using quantitative real-time reverse transcription PCR, western blotting, and immunohistochemistry. Cell migration, colony formation, wound-healing, and Transwell matrix penetration assays, revealed that upregulation of CEP55 promoted PANC cells proliferation, migration, and invasion in vitro, whereas knockdown of CEP55 attenuated it. In an in vivo murine model, CEP55 overexpression accelerated PANC cells tumourigenicity, together with upregulation of the protein levels of invasion-related proteins matrix metalloproteinase (MMP)2, MMP9, and proliferation-related protein Cyclin D1. Downregulation of CEP55 had the reverse effect. Moreover, the nuclear factor κB (NF-κB)/IκBα signalling pathway, which was activated in CEP55-transduced PANC cells and inhibited in CEP55-silenced PANC cells, contributed to CEP55-mediated PANC cell aggressiveness. This study provided new insights into the oncogenic roles of CEP55 and the mechanism by which the NF-κB pathway is hyperactivated in patients with PANC, indicating that CEP55 is a valuable prognostic factor and a potential therapeutic target in PANC.

Gastric cancer is one of the most common human malignant neoplasms, especially in China, its regulatory mechanism is important to develop new therapy approaches. miRNAs have been demonstrated to regulate gastric cancer progression. We found miR-532 was overexpressed in gastric cancer tissues and cells. Wound healing and transwell assay revealed that its overexpression promoted gastric cancer cell migration and invasion, its knockdown inhibited gastric cancer cell migration and invasion. Wnt/β-catenin antagonist naked cuticle homolog 1 (NKD1) was the target of miR-532, miR-532 inhibited NKD1 expression. TOP/FOP luciferase activity analysis suggested miR-532 also increased Wnt/β-catenin pathway activity. Overexpression miR-532 and NKD1 inhibited gastric cancer cell migration and invasion, consistent with miR-532 knockdown. These findings revealed miR-532 promoted gastric cancer cell migration and invasion through inhibiting NKD1 and activated Wnt/β-catenin pathway. We provide a potential target for gastric cancer therapy.

Rationale and Objectives Some deep pelvic abscesses are not accessible through anterior or lateral approaches because of the presence of organs and structures. The objective of this study was to assess the feasibility, safety, tolerability, and efficacy of a percutaneous presacral space approach by reviewing our clinical experience and the literature. Materials and Methods The outcomes of 12 patients, who have undergone computed tomography (CT)-guided percutaneous presacral space drainage, were retrospectively reviewed, including demographic, clinical, and morphological data in the medical records. Results From August 2010 to June 2015, 98 patients underwent CT-guided percutaneous drainage of pelvis abscesses in our institution. A percutaneous presacral space approach was adopted in 12 cases. The fluid collections were related to postoperative complications in nine patients (75%) and inflammatory or infectious intraabdominal disease in the remaining three patients (acute diverticulitis: n = 1; appendicitis: n = 1; Crohn disease: n = 1) (25%). The mean duration of drainage was 9.5 days (range 3–33). Escherichia coli was the most frequently present microorganism (in 50.0% of the all samples). No procedure-related complications were observed, either during or after the procedure. Drainage was successful in 10 patients (83.3%). Drainage failed in one patient because of massive anastomotic dehiscence. The other one died from pulmonary embolus 10 days after drainage. Conclusions When an anterior or lateral transabdominal approach is inaccessible, CT-guided transperineal presacral space approach drainage is a safe, well-tolerated, and effective procedure, except for patients with massive anastomotic dehiscence.

Long noncoding RNA (lncRNA) has been considered as potentially critical regulators in pancreatic ductal adenocarcinoma (PDAC). In this study, we prospectively investigate the effect and mechanism of lncRNA integrin subunit beta 2-anti-sense RNA 1 (ITGB2-AS1) on regulation of PDAC progression. The expression of ITGB2-AS1 and its target were analyzed by quantitative real-time polymerase chain reaction and in situ hybridization. 3-(4,5-Dimethylthiazol-z-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, wound healing, and transwell assays were used to investigate the influence of ITGB2-AS1 on cell proliferation, cell cycle, migration, and invasion, respectively. The interaction between ITGB2-AS1 and its target was determined via luciferase activity assay and RNA immunoprecipitation. The subcutaneous xenotransplanted tumor model was established and employed to detect the tumorigenic function of ITGB2-AS1, which was evaluated by western blot analysis, immunohistochemistry, and hematoxylin and eosin staining. The results showed that ITGB2-AS1 was elevated in both PDAC tumor tissues and cell lines, predicting a poor prognosis in PDAC patients. Knocking down of ITGB2-AS1 suppressed PDAC cell proliferation, invasion, and migration but induced cell apoptosis in vitro. Moreover, ITGB2-AS1 could target and inhibit the expression of miR-4319 and miR-4319-targeted and -suppressed serine/threonine kinase RAF1. ITGB2-AS1 promoted PDAC progression via inhibition of miR-4319. Interference of ITGB2-AS1 could suppress in vivo tumorigenic ability of PDAC via downregulation of RAF1. In conclusion, ITGB2-AS1 promoted PDAC progression via sponging miR-4319 to upregulate RAF1, suggesting the potential therapeutic target ability of ITGB2-AS1 in PDAC.

Trial Design: The aim of this study was to identify independent risk factors for post-pancreatoduodenectomy (post-PD) abdominal fluid collections (AFCs) and evaluate our management protocol on it. Methods: A retrospective analysis of consecutive 2064 cases who underwent PD over the past decade in 1 single center was conducted. The patients were divided into AFCs and non-AFCs group. Univariable and multivariate logistic regression analysis was performed to identify independent risk factors of AFCs. The AFCs group was compared with the non-AFCs group with respect to the incidence of postoperative outcomes. The characteristics of AFCs were further analyzed in terms of clinical manifestations. Results: Two thousand sixty-four cases with pancreaticoduodenectomy were recruited and 15% of them were found AFCs. Diameter of main pancreatic duct ≤3 mm was found to be an independent predictor of AFCs (P 800 mL (P

Background Modulation of cell surface expression of MHC class I chain-related protein A/B (MICA/B) has been proven to be one of the mechanisms by which tumor cells escape from NK cell-mediated killing. Abnormal metabolic condition, such as high glucose, may create a cellular stress milieu to induce immune dysfunction. Hyperglycemia is frequently presented in the majority of pancreatic cancer patients and is associated with poor prognosis. In this study, we aimed to detect the effects of high glucose on NK cell-mediated killing on pancreatic cancer cells through reduction of MICA/B expression. Methods The lysis of NK cells on pancreatic cancer cells were compared at different glucose concentrations through lactate dehydrogenase release assay. Then, qPCR, Western Blot, Flow cytometry and Immunofluorescence were used to identify the effect of high glucose on expression of MICA/B, Bmi1, GATA2, phosphorylated AMPK to explore the underlying mechanisms in the process. Moreover, an animal model with diabetes mellitus was established to explore the role of high glucose on NK cell-mediated cytotoxicity on pancreatic cancer in vivo. Results In our study, high glucose protects pancreatic cancer from NK cell-mediated killing through suppressing MICA/B expression. Bmi1, a polycomb group (PcG) protein, was found to be up-regulated by high glucose, and mediated the inhibition of MICA/B expression through promoting GATA2 in pancreatic cancer. Moreover, high glucose inhibited AMP-activated protein kinase signaling, leading to high expression of Bmi1. Conclusion Our findings identify that high glucose may promote the immune escape of pancreatic cancer cells under hyperglycemic tumor microenvironment. In this process, constitutive activation of AMPK-Bmi1-GATA2 axis could mediate MICA/B inhibition, which may serve as a therapeutic target for further intervention of pancreatic cancer immune evasion. Electronic supplementary material The online version of this article (10.1186/s13046-019-1209-9) contains supplementary material, which is available to authorized users.