Objective:To investigate the clinicopathological characteristics and prognostic factors of pancreatic cancer patients with liver metastasis.Methods:The clinical data of 67 pancreatic cancer patients with liver metastases who received first treatment in Department of Oncology of the First Affiliated Hospital of Naval Medical University between July 2012 and June 2016 were retrospectively analyzed. The relationship between patient survival time and the clinicopathological characteristics including patients' age, gender, tumor location, physical activity status score, tumor markers, number of distant metastatic organs, radiotherapy at the primary site, first-line chemotherapy regimen, number of cycles of first-line chemotherapy received, and liver metastases undergoing transcatheter arterial chemoembolization (TACE) was analyzed. Kaplan-Meier curves were plotted to reveal survival time in pancreatic cancer patients with liver metastases, and univariate and multifactorial COX proportional risk regression models were used to analyze independent prognostic risk factors for pancreatic cancer patients with liver metastases.Results:All patients were followed up until December 31, 2018, and all 67 patients died. The results of univariate analysis showed that patients with positive tumor marker, number of distant metastatic organs ≥2, number of cycles of first-line chemotherapy ≤2, no radiotherapy to the primary site and no TACE had shorter survival than those with negative tumor marker, one distant metastatic organ, number of cycles of first-line chemotherapy ≥3, with radiotherapy to the primary site and TACE, and all the differences were statistically significant (all P values <0.05). The results of multifactorial analysis showed that positive tumor markers ( HR=0.567, 95% CI 0.332-0.954, P=0.031), number of distant metastatic organs ≥2 ( HR=0.581, 95% CI 0.353-0.977, P=0.039), number of first-line chemotherapy cycles ≤2( HR=1.890, 95% CI 1.155-3.121, P=0.013) and primary foci without radiotherapy ( HR=0.414, 95% CI 0.231-0.732, P=0.002) were the independent prognostic risk factors for pancreatic cancer patients with liver metastasis. Conclusions:The prognosis of pancreatic cancer patients with liver metastasis is affected by multiple factors, among which positive tumor markers, more distant metastatic organs, no radiotherapy at the primary site and fewer first-line chemotherapy cycles are independent prognostic risk factors for pancreatic cancer with liver metastasis.

消化系统肿瘤是人类常见的恶性肿瘤，其中胃癌、结直肠癌、胰腺癌的发病率和病死率均较高。免疫治疗作为一种新 的治疗方法，正逐步成为多种肿瘤的有效治疗策略之一。CTLA-4和PD-1是通过不同机制负调控T细胞活化的关键免疫检查点 分子。针对这些免疫检查点抑制剂，已经显示出临床疗效，并已获美国FDA批准用于多种实体瘤的治疗。其中，纳武单抗在延长 晚期肝癌患者生存期方面超过了索拉非尼，派姆单抗在PD-L1阳性晚期食管癌有效率可达30%，纳武单抗与伊匹单抗联合治疗 dMMR/MSI-H晚期结直肠癌的客观缓解率达到55%。本文就近年来免疫检查点抑制剂在消化系统肿瘤治疗中的研究进展进行 综述。

Objective： ：To detect the distribution of gene mutations in pancreatic mucinous cystadenocarcinoma (PMCC) by highthroughput sequencing and to explore its clinical significance. Methods: Four cases of paraffin-embedded cancer tissues and paracancerous tissues from PMCC patients, who underwent surgical resection from January 2012 to December 2016, received NGS (next generation sequencing) examination using Illumina Hiseq 2500 platform. The characteristics of gene mutation in PMCC patients were analyzed with sequencing results and clinicopathological data. Results: Seven significantly mutated genes (SMGs) were detected in all four PMCC samples, namely KRAS, AHNAK2, MUC16, MUC17, MUC19, MUC3A and MUC4. Twenty-four SMGs were detected in 3 of the 4 samples, namely ADAMTS9, ALDH3B1, CARD14, CSMD3, MKI67, OR1N2, PKHD1, PLCE1, RTL1, SIGLEC12, CCDC168, CEP295, CUBN, DST, HRNR, LAMA5, OR10G4, OR2T4, PLEKHG4B, RP1L1, SLC15A5, SVEP1, TAS1R1 and TNRC18. KRAS-driven gene mutations were detected in all 4 samples, including K12 hot spot mutation in 3 cases and D33E non-hot spot mutation in 1 case. Conclusion: The high mutation of KRAS and MUC family in PMCC may be a potential target and biomarker for precise treatment of PMCC.

Due to the particularity of oncology,which is not included in undergraduate coume,most of the physicians gain the oncology knowledge in a fragmented way.The process of diagnosis and treatment of cancer needs to be concluded and systematization.According to characteristics of standardized training in the Department of Oncology,combined with the characteristics of internal medicine and oncology,we should increase the training of strengthening concept of multidisciplinary team cooperation for students,clinical thinking of cancer subjects,the training of transitional medical ideas and humanistic care knowledge with tumor characteristic,on the basis of strengthening skill training on Oncology.We should also encourage the general physicians of non cancer majors to participate in the standardized training of cancer specialties and explore more teaching methods and forms,as well as the positive significance of cross professional training.

Over the last few decades progress in the field of stem cell biology makes a new kind of 3-D human cells in vitro culture techniques available, which is termed organoids because of its space structure similar to organs. Organoids derived from tumor tissues largely retained the biological characteristics of tumor tissue in vivo, and both the advantages of low cost and easy operation make up the defects of the previous conventional cancer models. Organoids can also be used for translational medical research, formulating individual therapeutic schedules including drug sensitive tests, and combining with many kinds of technologies. This article focus on the application and prospectsoforganoidsintumorresearch.

Objective: To construct a hsa-microRNA-224(miR-224) lentiviral expression vector and to establish pancreatic mucinous cystadenocarcinoma MCC1 cell line with stable miR-224 over-expression. Methods: Pri-miR-224 gene fragment was designed and amplified by quantitative real-time polymerase chain reaction (qRT-PCR), and then loaded into GV369 lentiviral vectors (GV369-miR224) by gene recombination technology. GV369-miR-224 lentivrial expression vectors were then identified by PCR and DNA sequencing. The GV369-miR-224 vector fluid was then used to infect pancreatic mucinous cystadenocarcinoma MCC1 cell line to establish the MCC1 cell line stably over-expressing miR-224. The transfection efficiency of GV369-NC and GV369-miR-224 was observed under fluorescence microscopy; and the expression levels of miR-224 in MCC1, GV369-miR-224-MCC1 and GV369-NC-MCC1 cell lines were detected by RT-PCR. Results: The GV369-miR-224 lentiviral vectors were successfully constructed. GV369-miR-224-MCC1 and GV369-NC-MCC1 cells all emit green fluorescence under the fluorescence microscope. The expression level of miR-224 in GV369miR-224-MCC1 cell group was significantly higher than that in negative control GV369-miR-224-MCC1 group and blank control MCC1 cell group (23.45±1.94， 1.46±0.1 and 2.11±0.38, P<0.01), however, there was no significant difference between the two control groups (P>0.05). Conclusion: A pancreatic mucinous cystadenocarcinoma MCC1 cell line with stable miR-224 over-expression was successfully established, and this will provide a new cell model for exploring the function and pathogenesis of miR-224 in pancreatic mucinous cystadenocarcinoma.

Objective To investigate the effect of apatinib on the proliferation,apoptosis and migration of pancreatic cancer cell line AsPC-1 in vitro.Methods Pancreatic cancer AsPC-1 cells were treated by apatinib in different concentrations.Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry,and the effect of apatinib on cell migration ability was observed by wound healing assay.Results In control and 10,20,30,40 and 50umol/L apatinib treatment group,the inhibitory rates of AsPC-1 cells were 0,(1.45 ±0.68)％,(16.92±0.70)％,(23.84±0.84)％,(34.35±1.55)％ and (37.33± 0.81) ％,respectively.Cell proliferation was obviously inhibited by apatinib as the concentration increased,and the differences were statistically significant (P ＜ 0.05).In control and 20,40 umol/L apatinib treatment group,the apoptotic rates were (9.44 ± 0.18) ％,(16.62 ± 0.19) ％ and (25.42 ± 0.41) ％,respectively.Number of apoptotic cells was obviously increased by apatinib as the concentration increased,and the differences were statistically significant (P ＜ 0.05).In control and 20,40 umol/L apatinib treatment group,the migration ability was (29.5 ± 0.7) ％,(17.4 ± 0.9) ％ and (6.6 ± 0.5) ％,which was greatly decreased as the concentration increased,and the differences were statistically significant (P ＜ 0.05).Conclusions Apatinib can effectively inhibit the proliferation and migration of pancreatic cancer AsPC-1 cells and induce apoptosis.

Objective To silence the beclin1 gene expression by using RNA interference technology in AR42J rat pancreatic acinar cells,and build a stable AR42J line silencing beclin1.Methods Three kinds of shRNA targeting rat beclin1 mRNA and negative control shRNA were designed and synthesized,and were inserted into the plasmids GV112,respectively.The recombinant plasmids were named as p-sh-Beclin1-1,psh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC.Lipo3000 was used to transfect the recombinant plasmid into AR42J cells,the expression of beclin1 mRNA were detected by RT-PCR to screen for the most efficient silencing plasmid,and then it was packaged into lentiviral (LV).AR42J cells were infected with LV and screened by puromycin.Beclin1 mRNA and protein expression was determined by RT-PCR and Western blot.Results The recombinant plasmid was confirmed by agarose gel electrophoresis and sequencing showed that shRNA sequences were in line with expectations.The beclin1 mRNA inhibition rates of AR42J cells after p-sh-Beclin1-1,p-sh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC transfection were (17.8 ± 4.0) ％,(30.6 ± 2.8) ％,(45.8 ± 7.7) ％,(7.0 ± 11.8) ％,respectively.The inhibition rates of three p-sh-Beclin1 transfection cells were significantly higher than that in non-transfection cells,and the difference was statistically significant (P ＜ 0.05).While the inhibition rate of p-shRNA-NC transfection cells was not significantly different from that of non-transfection cells,p-sh-Beclin 1-3 with highest rate of inhibition was packaged by LV,and infected AR42J cells,then puromycin was applied to screen,inhibition rate of beclin mRNA expression in LV infection cells was (86.1 ± 1.2) ％,and the protein expression inhibition rate was (87.9 ± 2.8) ％,and the difference between infection and non-infection groups was statistically significant (P ＜ 0.05).Conclusions The stable AR42J line silencing beclin1 is successfully established,which can provide a new cell model for future research of the role of beclin1 in the pathogenesis of acute pancreatitis.

Objective To investigate the changes and significance of autophagy in rats with experimental acute necrosis pancreatitis (ANP).Methods According to method of random number,18 rats were randomly divided into control group,ANP group,ANP+rapamycin (RAP) group.The ANP rat model was established by intraperitoneal injection of 20％ L-arginine.The rats of ANP+RAP group were intraperitoneal injected with RAP 1.2 mg/kg at 30 minutes before modeling.The rats of control group were intraperitoneal injected with 0.9％ NaCl solution.The blood was drawed from the hearts nine hours after modeling for subsequent experiments.Serum levels of trypsinogen activation peptide (TAP),interleukin (IL-1),IL-6 and tumor necrosis factor (TNF) α were measured with enzyme-linked immunosorbent assay.The pancreatic tissues were pathologically scored.Autophagy-related structures in rat pancreatic acinar cells were observed by transmition electron microscopy.The expression of autophagy marker microtuble assciated protein 1 light chain 3 (LC3)-Ⅱ and Beclin-1 at mRNA and protein level were measured by quantitative real-time polymerase chain reaction (qRT-PCR),Western bloting and immunohistochemistry.The single factor analysis of variance was used for mean comparison among groups.Results A rat model of ANP was successfully established.Histopathological score of pancreas acinar cell necrosis of ANP+RAP group (2.19±1.38) was higher than that of ANP group (0.97±0.68),and the difference was statistically significant(F=33.75,P＜0.05).The results of Western blotting indicated that the protein expression of LC3-Ⅱ and Beclin-1 in ANP group (35.25±2.68 and 49.40±5.28)were higher than those in control group (1.54±0.16 and 0.78±0.06),furthermore the expressions in ANP+RAP group(123.53±3.21 and 76.41±3.80) were higher than those in ANP group,and the differences were statistically significant(F=2 045.54,326.87,both P＜0.01).Immunohistochemistry results also indicated that the LC3Ⅱ and Beclin-1 expression at protein level of ANP+RAP group (7 570.63±4 357.67 and 3 418.09±2 035.78) were higher than those of ANP group (1 926.53±1 414.44 and 536.11±403.10),and the differences were statistically significant (F=39.83,41.58,both P＜0.01).The expression of Beclin-1 at mRNA level of ANP group (107.12±29.10) was statistically higher than that of control group(7.01 ±3.39),and the difference was statistically significant (F=3.61,P＜0.05),but the expression of ANP+RAP group (97.63 ± 65.38)was no significant difference compared with ANP group.However,the expression of LC3-Ⅱ at mRNA level of ANP+ RAP group (4.37 ± 1.67) was statistically higher than that of ANP group (1.76 ± 1.59),and the difference was statistically significant(F=16.10,P＜0.05),but the expression of ANP group was no significant difference compared with control group (1.51 ±0.95).The result of electron microscopy showed that autophagy related structures increased in ANP group compared with that of control group,which of ANP+RAP group was more.The serum levels of TAP,IL-1 and IL-6 of ANP + RAP group were (36.47 ± 1.71) pmol/L,(122.88± 26.67) pg/mL and (107.39±13.95) pg/mL,which were all higher than those of ANP group ((25.63 ± 6.05) pmol/L,(98.06 ±9.29) pg/mL and (86.16± 7.20) pg/mL),and the differences were statistically significant (F=116.71,50.45,79.67; all P＜0.01).There was no significant difference in TNFα between ANP+ RAP group ((140.80±60.82) pg/mL) and ANP group ((105.23±6.95) pg/mL,F=14.76,P＞0.05).Conclusions Autophagy increased in rats with ANP.Promoting autophagy could significantly activate trypsinogen,aggravate pancreatic injury and increase inflammation reaction,which indicated that autophagy might involve in the pathogenesis of ANP through trypsinogen activation.

Background:Damage of intestinal mucosal barrier is a key factor in the development and progress of acute pancreatitis(AP),and is closely related with the prognosis of the disease. Aims:To investigate the protective effect and possible mechanism of mucoprotective agent teprenone on intestinal mucosal barrier in rats with experimental AP. Methods:Forty-five adult male Sprague-Dawley rats were randomly divided into normal control group(n = 5),AP model group(n = 20)and teprenone treated group(n = 20). AP model was established by subcutaneous injection of cerulein at abdominal wall. Rats in treated group were intervened with teprenone intragastrically before and after model establishment. ELISA was used for measurement of serum interleukin-1(IL-1),IL-6,tumor necrosis factor-α(TNF-α)and amylase;histopathological and ultrastructural changes of small intestinal mucosa were observed by light microscope and transmission electron microscope;Western blotting was used to detect the expressions of tight junction protein occludin and ZO-1. Results:Serum levels of IL-1,IL-6,TNF-α and amylase in AP model group were significantly higher than those in normal control group(P < 0. 05),accompanied by necrosis and exfoliation of small intestinal villus,widening of intercellular tight junctions and downregulation of occludin and ZO-1 expression. While in teprenone treated group,serum levels of proinflammatory cytokines and amylase were significantly decreased as compared with AP model group(P < 0. 05),the villus of small intestine remained intact,and dense tight junctions were observed. Expressions of occludin and ZO-1 in teprenone treated group were upregulated. Conclusions:Teprenone may protect against intestinal mucosal barrier injury in AP model rats by upregulating tight junction protein expression.