Pancreatic cancer is characterized by nerve invasion and a high mortality rate， and its pathological process depends on the complex interaction network between tumor and the nervous system. Based on the concept of “pancreatic cancer neuroecology”， this article analyzes the mechanism of action of peripheral motor nerve， sensory nerve， and central nerve in tumorigenesis， pain regulation， and cachexia formation and emphasizes the synergistic regulatory role of immune cells， Schwann cells， and extracellular matrix in the microenvironment of perineural invasion. At the same time， this article further elaborates on the metabolic interaction and chemotaxis between neuraxis and tumor， the effect on promoting chemotherapy resistance， and the dynamic relationship between neuroplasticity and tumor adaptability. In clinical practice， this article summarizes the key value of perineural invasion in prognostic evaluation， preoperative evaluation， and the selection of surgical strategy. In addition， this article reviews the basic research advances in the biomarkers and potential targets associated with perineural invasion in pancreatic cancer and points out the limitations of current model and transformation research. In the future， systematically analyzing the nerve-tumor-immune network and targeting its key nodes may provide multi-dimensional strategies and new breakthroughs for the precise intervention of pancreatic cancer， the reversal of drug resistance， and the relief of symptoms.

Objective:To determine the expression of circular RNA-SEC31A(circSEC31A) in pancreatic cancer and investigate the effects on the invasion and migration of pancreatic cancer cells and the underlying molecular mechanism.Methods:Differentially expressed circRNAs between pancreatic cancer cells (BXPC-3, PANC1, CaPan-2, SW1990) and human normal pancreatic cells (HPDE) were identified by qRT-PCR. Then, two cell lines with high circSEC31A expression were selected to conduct next experiments. According to the sequence of the back-splicing site in circSEC31A, siRNAs for downregulation of circSEC31A were designed and transfected by liposome to silence circSEC31A in pancreatic cancer cells, and grouped as followed siR-circSEC31A#1 and siR-circSEC31A#2. Meanwhile, siR-NC group transfected with non-specific siRNA served as control. Transwell assays and wound healing assays were operated to evaluate the functional role of circSEC31A on the invasion and migration of pancreatic cancer cells. RNA Pull-down assay with circSEC31A probe and oligo control probe was used to screen the miRNA combining with circSEC31A and the effects of miRNA on cell invasion and migration of pancreatic cancer cells were validated. The effect of miR-200c-3p and circSEC31A silencing on the expression of PDK1 mRNA was identified by qRT-PCR. The protein expression of PDK1, downstream Akt and p-Akt after circSEC31A silencing was verified by Western blotting assays.Results:The relative expression level of circSEC31A in HPDE (1.000±0.120) was obviously lower than that in BXPC-3 (1.920±0.130), SW1990 (2.93±0.528), PANC1 (4.557±0.692) and CaPan-2 (5.247±0.194), and all the differences were statistically significant ( P<0.001). Compared with the PANC1 siR-NC group (1301.3±94.6) and CaPan-2 siR-NC group (1835.0±70.1) per 100 high power field, transwell assays showed that the numbers of invasive pancreatic cancer cells was highly decreased in PANC1 siR-circSEC31A#1 group (727.3±92.9), siR-circSEC31A#2 group (792.0±18.1), CaPan-2 siR-circSEC31A#1 group (718.0±90.6), siR-circSEC31A#2 group (692.7±84.8). Wound healing assays showed that silencing circSEC31A decreased the wound healing rate of pancreatic cancer cells in PANC1 siR-circSEC31A#1 group (20.667±3.215)%, siR-circSEC31A#2 group (20.000±4.583)%, CaPan-2 siR-circSEC31A#1 group (28.000±8.185)%, siR-circSEC31A#2 group (29.667±5.686)%, compared with the PANC1 siR-NC group (55.000±4.359)% and CaPan-2 siR-NC group (69.000±3.606)%. RNA Pull-down assays showed that compared with PANC1 oligo probe group (1.000±0.091) and CaPan-2 oligo probe group (1.000±0.153), miR-200c-3p was significantly enriched in the PANC1 circSEC31A probe group (2.237±0.175) and CaPan-2 circSEC31A probe group (2.166±0.156). Compared with PANC1 siR-NC group (939.3±57.0) and CaPan-2 siR-NC group (786.7±51.5) per 100 high power field, the numbers of invasive pancreatic cancer cells were up-regulated in PANC1 siR-miR-200c-3p group (1206.0±99.1) and CaPan-2 siR-miR-200c-3p group (1838.0±105.7), while the low numbers of invasive pancreatic cancer cells were observed in PANC1 siR-miR-200c-3p+ siR-circSEC31A group (932.7±116.4) and CaPan-2 siR-miR-200c-3p+ siR-circSEC31A group (785.3±58.8). Compared with PANC1 siR-NC group (1.000±0.103) and CaPan-2 siR-NC group (1.000±0.107), the relative expression of PDK1 mRNA in PANC1 siR-miR-200c-3p group (1.898±0.159) and CaPan-2 siR-miR-200c-3p group (2.102±0.337) was upregulated. Furthermore, the expression of PDK1 mRNA was decreased in the siR-miR-200c-3p+ siR-circSEC31A group (0.980±0.070, 1.015±0.079). Western blot assays showed that the expression of PDK1 protein in PANC1 siR-NC group, siR-circSEC31A#1 group, siR-circSEC31A#2 group was 0.767±0.086, 0.281±0.191, 0.333±0.062 and in CaPan-2 siR-NC group, siR-circSEC31A#1 group, siR-circSEC31A#2 group was 0.712±0.038, 0.353±0.061, 0.308±0.018. The expression of p-Akt protein in PANC1 siR-NC group and siR-circSEC31A group was 0.741±0.050, 0.114±0.027, 0.139±0.041. In addition, p-Akt protein expression in CaPan-2 siR-NC group and siR-circSEC31A group was 0.823±0.052, 0.141±0.045, 0.280±0.089. PDK1 and p Akt expression in siR circSEC31A group was obviously lower than those in sir NC group. All the differences between either groups above were statistically significant ( P<0.05). Conclusions:circSEC31A is upregulated in pancreatic cancer cells, which facilitates the invasion and metastasis of pancreatic cancer cells via miR-200c-3p/PDK1/Akt signaling pathway, supporting that circSEC31A may function as a new diagnostic and therapeutic target for pancreatic cancer patients.

Objective To explore the effects of hepatocyte nuclear factor 1 homeobox A(HNF1A) on drug resistance of PANC1 cells to gemcitabine plus abaraxane and explore the potential mechanism. Methods 78 pancreatic cancer patients with locally advanced or distant metastasis who received gemcitabine plus abaraxane chemotherapy after surgery in Biliary and Pancreatic Surgery Department of Sun Yat-sen Memorial Hospital from March 2012 to May 2017 were enrolled. qPCR was used to detect HNF1A mRNA levels in pancreatic cancer tissue. The patients were divided into high-expression group ( n=39 ) and low-expression group (n=39) according to the median expression level of HNF1A, and the correlation of HNF1A expression with cancer clinicopathologic parameters and survival was analyzed. qPCR was used to detect HNF1A mRNA of 3 drug-sensitive cell lines (BxPC-3, CFPAC-1 and L3. 6pl) and 4 drug-resistant pancreatic cancer cell lines (PANC1, MIA PaCa-2, Hs766T and Mpanc96). Lentivirus with plasmids carrying HNF1AcDNA infection was used to establish HNF1A overexpressing PANC1 cells ( HNF1A group), and lentivirus with empty plasmids were used to infect PANC1 cells to construct the control group. The mRNA and protein expression of HNF1A and ATP binding cassette transporter family ABCC1 in HNF1A group and control group were measured by qPCR and Western Blot, respectively. The half inhibition concentration ( IC50 ) of gemcitabine plus abaraxane was detected by MTT, and cell apoptosis was examined by flow cytometry. Results Pancreatic cancer patients with high HNF1A expression had a better overall survival than those with low HNF1A expression (17. 9 months vs 12.4 months), and the difference was statistically significant (P<0.001). HNF1A low expression in pancreatic cancer tissue was significantly associated with advanced TNM stage, perineural invasion ( PNI) and short overall survival. The expression level of HNF1A was significantly down-regulated in drug-resistant PANC1 cells compared to drug-sensitive BxPC-3 cells by an average fold change of 6. 73, and the difference was statistically significant ( P<0. 001 ). In HNF1A group, the mRNA and protein levels of ABCC1 were significantly decreased compared with those in control group (0. 012 ± 0. 004vs 0. 047 ± 0. 008,0. 281 ± 0. 040 vs 0. 832 ± 0. 046,P=0. 003,P <0. 001). IC50 of HNF1A group to gemcitabine plus abraxane was decreased compared with that of control group [(26. 31 ± 2. 91)μmol/L vs (72. 63 ± 4. 07) μmol/L], and the cell apoptosis rate of HNF1A group was increased compared with that of control group [(40. 18 ± 1. 64)％ vs (21. 31 ± 1. 98)％], and the differences were statistically significant (P<0. 01). Conclusions HNF1A may induce resistance of pancreatic cancer cell to gemcitabine plus abraxane by downregulating ABCC1.

Pancreaticoduodenectomy is the only possible surgical procedure for the cure of pancreatic head carcinoma. With the development of minimally invasive surgery, the minimally invasive surgical treatment of pancreatic head carcinoma has become more mature, and an increasing number of grade A tertiary hospitals have reported the minimally invasive surgery for pancreatic head carcinoma. With reference to related articles and experience of our center for pancreatic surgery, this article elaborates on the current status of minimally invasive surgical treatment of pancreatic head carcinoma and points out the perspectives and development directions of minimally invasive treatment of pancreatic head carcinoma.

Objective To explore the clinicopathological characteristics of pancreatic neuroendocrine neoplasms ( P-NENs) and the prognosis .Methods The clinicopathological data of 72 patients with P-NENs in Sun Yat-Sen Memorial Hospital from December 2011 to September 2017 were analyzed retrospectively , and the gender, age, tumor size, local infiltration, neural invasion and distant metastasis were collected .According to the diagnostic criteria of P-NENs ( Chinese edition 2013 ) , tumor classification was determined by histological mitotic figure count and cell proliferation index ( Ki67 ) proliferation activity .Expressions of Syn , CgA, NSE and CD56 were determined by immunohistochemical staining .Results Seventy-two patients with P-NENs were enrolled, including 33.3％male(n=24) and 66.6％ female (n=48), and the median age was 56 years old (range:12-76).24 patients (33.3％) were NET G1,44 patients (61.1％) were NET G2 and 4 patients (5.6％) were NET G3.The mean size of tumor was 38.2 ±19.2 cm3 ( range:0.1-371 cm3 ). Immunohistochemistry staining positive rates for Syn , CgA, NSE and CD56 were 98.6％, 95.8％, 88.9％and 90.3％ respectively.Tumor classification was correlated with tumor size , local infiltration, perineural infiltration and distant (liver) metastasis (all P<0.05).There was no statistically significant differences on positive rate of Syn, CgA, NSE and CD56 among G1, G2 and G3.Conclusions Tumor classification, size, local invasion , perineural infiltration and distant ( liver ) metastasis were all important clinicopathological features and prognostic factors of P-NENs.Syn, CgA and Ki67 were essential immunohistochemical markers to diagnose P-NENs.

We compared the ways of deproteinization for crude polysaccharides of Coprinus comatus, and finally selected Sevage method as the optimal method. Two main fractions of Ccp-I-A and Ccp-I-B were obtained after DEAE-52 cellulose and Sephadex G-200 chromatography, both were white-floc, soluble in water, insoluble in absolute ethyl alcohol, acetone and other organic solvents. Additionally, Fehling reagent, CTAB, Sulphuric acid-carbazole, I-KI and FeCl₃ reaction were all negative. GC analysis showed Ccp-I-A was composed of mannitose, glucose and galactose in molar ratios of 2.03:9.52:1, whereas Ccp-I-B was composed of fucose and galactose with molar ratios of 1:5.21. Antioxidant activity test showed that Ccp-I-A and Ccp-I-B had good scavenging abilities on DPPH and ·OH. Compared to Ccp-I-B,the scavenging activity of Ccp-I-A was much stronger, and the scavenging rate could reach 72.1% and 55.3% respectively when the concentration was 300 μg/mL.

Objective To investigate the assessed value of tumor-associated macrophages ( TAMs ) and KIT expression for liver metastasis in pancreatic neuroendocrine tumors (PNETs) and patients′outcome. Methods A total of 79 patients who underwent surgical resection and pathologically diagnosed as PNETs in the Department of Hepatopancreatobiliary Surgery in Sun Yat-sen Memorial Hospital from January 1995 to May 2015 were enrolled.The immunohistochemical staining of CD68 and KIT were detected and the correlations with clinicopathological factors were analyzed.Results Of 79 PNETs cases, CD68 and KIT in tumor tissue were overexpressed in 30(38%) and 35(44.3%) cases, respectively.CD68 overexpression was associated with tumor infiltration ( P<0.001 ), AJCC stage 7 ( P<0.001 ), liver metastasis ( P<0.001 ) and early recurrence (P=0.019).Patients with low CD68 level had significantly better survival than those with high CD68 expression ( P=0.0002 ).KIT overexpression was correlated with WHO 2010 and AJCC stage 7 (P<0.001;P=0.002), nonfunctional status of the tumor (P=0.002) and liver metastasis (P=0.026). The survival period of patients with low KIT expression was greatly longer than those with high KIT level (P=0.0013).CD68 and KIT co-overexpression was observed in patients with tumor invasion (P<0.001), advanced WHO and AJCC stage (both P<0.001) and better prognostic survival (P=0.0057).Multivariate analysis showed that CD68 overexpression (HR:2.9;95%CI:1.16~7.23;P=0.033) was an independent prognostic factor for PNETs.Conclusions CD68 and KIT overexpression is correlated with advanced disease stage, higher risk for liver metastasis and worse survival.CD68 is an independent prognostic factor for PNETs.

OBJECTIVETo explore whether hepatitis C virus core protein (HCV C) regulates the expression of NFAT1 to participate in the progression and malignant biological behavior of intrahepatic cholangiocarcinoma cells.

METHODSThe recombinant plasmid pEGFP-N(3)-HCV C and the empty vector pEGFP-N(3) were cotransfected with enhanced green fluorescent protein (EGFP) into RBE cells using liposome. Real-time PCR and Western blotting were used to examine the expression of NFAT1 mRNA and protein in the transfected RBE cells. MTT assay was used to evaluate the changes in the cell proliferation, and the cell cycle changes were analyzed by flow cytometry.

RESULTSHCV C transfection significantly enhanced the expressions of NFAT1 mRNA and protein in RBE cells (P<0.05) and promoted the progression of cell cycle into G(2)/M phase to accelerate the cell proliferation.

CONCLUSIONTransfection with HCV C gene up-regulates NFAT1 expression and promotes the cell cycle progression and proliferation of intrahepatic cholangiocarcinoma cells, suggesting the involvement of HCV C in the progression of intrahepatic cholangiocarcinoma.

Bile Duct Neoplasms ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma ; pathology ; Gene Expression ; Humans ; NFATC Transcription Factors ; genetics ; Plasmids ; Transfection ; Viral Core Proteins ; genetics

ObjectiveTo review our experience in the diagnosis and surgical treatment of solid pseudopapillary neoplasm (SPN) of the pancreas which can be used as a reference for other doctors to avoid misdiagnosis and to provide a better treatment.MethodThe clinical,laboratory,radiological,pathological and operative data of 24 patients with SPN of the pancreas operated between February 2001 to December 2009 were collected and retrospectively analyzed.Results23 of 24 patients were female and the mean age was 31 years.The most common clinical presentations were vague abdominal pain and abdominal mass.In most cases,abdominal imaging showed a solid or a solid-cystic mass in the tail or head of pancreas.All patients received surgery.20 of 22 patients who received curative resection were alive with no evidence of tumour recurrence.One patient who had a R1 resection died 42 months after surgery.The remaining patient was alive after a second operation.ConclnsionsSPN of the pancreas is a tumour with low malignancy.A correct diagnosis of SPN of the pancreas is made on its clinical,radiological and histopathological characteristics.Radical surgical resection is the treatment of choice.For patients with an advanced disease,palliative resection is beneficial.

Nuclear factor-kappa B (NF-κB) is a key regulator in epithelial-mesenchymal transition (EMT) of cancer cells.EMT of cancer,one of the reasons of drug resistance,enhances the infiltration and distant metastases ability of cancer cells.Recent researches show that Snail,Slug,Twist and Zeb play important roles in regulating EMT of cancer cells.Drugs and targeted therapies that inhibit NF-κB activities can reverse the EMT of cancer cells.NF-κB may become an effective therapeutic target in cancers in the future.