Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)and matrix metalloproteinase-9(MMP-9) in MIA PaCa-2 cells blocked by AS-ODN cultured in hypoxia.Methods Heparanase(Hpa) expression of MIA PaCa-2 cells was blocked by AS-ODN and cultured in hypoxia.The expression of MMP-2 and MMP-9 mRNA and proteins in cell lysate was evaluated by RT-PCR and Western blot respectively,and the enzymatic activities of MMP-2 and MMP-9 in supernatants were detected by gelatinase activity assay.Results Hypoxia stimulated mRNA and protein expression of MMP-9 in cultured MIA PaCa-2 cells and elevated at 6h,12 h(P ＜0.05)and 24 h(P ＜ 0.01).When Hpa expression was inhibited by AS-ODN,the expression of MMP-9 mRNA and protein as well as the gelatinase activity in supernatant decreased dramatically at 12 h and 24 h,especially at 24h(P ＜0.01),however,no significant difference of MMP-2 expression and gelatinase activity was observed after AS-ODN transfection.(P ＞ 0.05).Conclusion In hypoxia,MMP-9 expression,either mRNA or protein in cultured MIA PaCa-2 cells,increased gradually accompanied with elevated gelatinase activities.When the heparanase expression was inhibited,the MMP-9 mRNA and protein,as well as the gelatinase B activity in supernatant,were decreased dramatically at 12h and 24h,however,no significantly differences of MMP-2 expression and gelatinase A activity were observed after the AS-ODN transfection.

OBJECTIVE To investigate the expression of SleX and CD24 in nasal inverted papilloma and its pathologic features.METHODS HE staining were conducted to study the pathologic features in specimens of 11 cases with nasal inverted papilloma. Further,immunohistochemistry stain for SleX and CD24 were performed in total specimens.RESULTS One case(9.1%) was diagnosed as severe atypical hyperplasia but tumor cells did not invaded basal membranes.SleX staining located at cell membranes. Positive SleX staining was found in 9 specimens (81.8%) and 1 normal nasal epithelium (16.7%).CD24 staining located in cytoplasm.Positive CD24 staining was found in 8 specimens of nasal inverted papilloma (72.7%). CD24 was negative in nasal epithelial cells and only a few lymphocytes were positive.CONCLUSION Some cases of nasal inverted papilloma are diagnosed with severe atypical hyperplasia.Most of cases express CD24,so nasal inverted papilloma may be a borderline tumor.Expression of SleX and infiltration of inflammatory cells suggest that nasal inverted papilloma may be related to inflammatory reactions.

Objective To establish a model of rat orthotopic liver transplantation and investigate the relationship among cold preservation time, activation of nuclear transcription factor-κB (NF-κB) and donor preservation injury after liver transplantation. Methods Orthotopic liver transplantation was performed in Wistar rats which were randomly divided into the following groups according to the different duration of liver cold storage in UW solution: group A (sham operation, n=10), group B NF-κB in liver before and after transplantation was measured by electrophoretic mobility shift assays; protein expression of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) in the liver was measured by immunohistochemistry; the serum TNFα and IL-1β, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hepatic cell apoptosis were examined. Results With extended cold storage duration, the activity of NF-κB in the donor liver increased (P<0.05, group D vs. groups A, B and C). TNF-α and IL-1β levels also increased (P<0.05, group D vs. groups A, B and C). Donor liver reperfusion injury was gradually aggravated with the prolonging of graft cold preservation. Both the serum TNF-α and IL-1β levels increased highly (P<0.05 group A vs. groups B, C and D),NF-κB in the liver significantly increased (P<0.05, group A vs. groups D, B and C) with gradual prolonging of graft cold preservation time. The serum ALT and AST level and apoptosis index level elevated greatly (P<0.05, group A vs. groups D, B and C). Conclusion NF-κB of donor liver was activated inductively in cold preservation phase and activated explosively in reperfusion phase. The longer cold preservation time was, the higher NF-κB level in donor liver became. NF-κB led to the expression of TNF-α and IL-1β in donor liver. Inflammatory factors are one of the most important mechanisms for the donor liver injury after liver transplantation.

Objective To study the regulation of heparanase expression by hypoxia and its correlation to the invasiveness of tumor cells. Methods BxPC-3 cells were cultured in hypoxia in vitro and the heparanase mRNA and protein expression were detected by reverse transcriptional polymerase reaction chains (RT-PCR) and western blot respectively. Matrigel invasion assay was used to observe the invasive abilities of tumor cells in hypoxia and in the status of heparanase was inhibited by antisense oligodeoxynucleotide (AS-ODN) targeting to the heparanase gene promoter. Results In normoxia, there was a relatively low level of heparanase mRNA and protein expression in cultured BxPC-3 cells. In hypoxia, heparanase expression, mRNA and protein which expressed consistently, were inhibited slightly at 3h and upregulated significantly at 6h, 12h, 24h and 48h. When the heparanase expression was inhibited by AS-ODN, the heparanase mRNA and protein maintained low in hypoxia, however, the nonsense oligodeoxynucleotide (NS-ODN) did not block upregulation of heparanase expression. In matrigel assay, after 48h incubation, number of BxPC-3 cells that penetrated the Matrigel-coated filter of transwell chamber was increased 96.2% in hypoxia (P<0.01), the Hpa AS-ODN (400 nmol/L) inhibited the invasive cells by 37.2% (P<0.05). Conclusions BxPC-3 cells invasion ability is enhanced by hypoxia through upregulation of heparanase mRNA and protein expression in BxPC-3 pancreatic cancer cell lines.

Intestinal transplantation is the only treatment option for irreversible intestinal failure and parenteral nutrition failure.Although the progress of immunosuppressive therapy and the improvement of surgical techniques have improved the prognosis of intestinal transplantation,it is still unsatisfactory compared with other solid organ transplantation,mainly due to the fact that the intestinal mucosa is susceptible to damage due to ischemia.The basic pathophysiological process of ischemia-reperfusion injury(IRI) is that ischemia/reperfusion induces the production of reactive oxygen species (ROS),inflammation and cell apoptosis,leading to tissue damage.In recent years,the pathophysiology of ischemia-reperfusion injury in intestinal transplantation has made some new progress,providing more possibilities for the prevention and treatment of intestinal transplantation IRI.Although most of them are animal experimental results,they have important reference value for clinical application.

Objective To compare salvage liver transplantation (SLT) with othotopic liver transplantation (OLT) in treatment of hepatocellular carcinoma.Methods A systematic literature search of PubMed,Embase,Cochrane Library,CBM,CNKI and Wanfang Med Online was performed from their dates of establishment to October 2017.The results were screened,data extracted and then analyzed with Stata 14.Results 23 studies with 4 161 patients were selected,including 579 patients in the SLT group and 3 582 patients in the OLT group.Compared with OLT,SLT was associated with a longer operative time (SMD =0.56,95％CI:0.29～0.83),higher intraoperative blood loss (SMD=1.56,95％CI:0.63～2.49),an increased risk of postoperative bleeding (OR =1.84,95％CI:1.08 ～ 3.14),a poorer overal survival rate (HR =1.29;95％CI:1.11～1.49) and disease free survival rate (HR=1.88;95％CI:1.26～2.81).The differences were all significant (all P＜0.05).The biliary complications (OR=1.25;95％CI:0.79～1.98),vascular complications (OR=1.41;95％CI:0.69～2.89),sepsis (OR=1.10;95％CI:0.60～ 1.99),acute rejection (OR =1.25;95％ CI:0.69 ～ 2.28) and perioperative mortality (OR =1.60;95 ％ CI:0.94 ～ 2.70) rates were not significantly different (all P＞0.05).Conclusions OLT is a better treatment strategy for patients with transplantable hepatocellular carcinoma (HCC) compared with SLT.However,severe organ limitation,and feasibility and safety of surgery make SLT a better option for patients with HCC recurrence after liver resection.

Objective:To Eveluate the safty and clinical efficacy of combined laparoscopic spleen-preserving distal pancreatectomy and autologous islet transplantation in the treatment of solid pseudopapillary neoplasm.Methods:A 22 years old solid pseudopapillary neoplasm female patient who underwent distal pancreatectomy and an autologous islet transplantation at Tianjin First Central Hospital, clinical date for 6 months follow up was collected and analyzed.Results:The patient was well recovered after surgery, and during the post-operative follow up, the fasting blood glucose was 5.72 mmol/L, HbA1c was 6.1%, remained insulin independent, the liver function was kept well.Conclusions:Combined Laparoscopic spleen-preserving distal pancreatectomy and autologous islet transplantation can effectively prevent diabetes after distal pancreatectomy.

Objective:To investigate the effect of miR-199a on interstitial transdifferentiation (EMT) of human renal tubular epithelial cells stimulated by high glucose.Methods:Human renal tubular epithelial cells were cultured in vitro and divided into low glycemic group (LG), high osmotic group (HM) and high glycemic group (HG) according to different glucose concentrations. They were divided into miR-199a inhibition group (miR-199ai group), negative control group (miR-INC group) and liposome group (Mock group) according to whether they were transfected with miR-199a inhibitors (miR199ai). They were divided into si-hypoxia-inducible factor-1α (si-HIF-1α) group, miR-199ai group, miR-199ai+ si-HIF-1α group and negative control group (NC group) according to whether they converted to si-HIF-1α and miR-199ai. The expressions of miR-199a, HIF-1α, fibronectin (FN) and α-smooth muscle actin (α-SMA) in different groups of cells were detected by real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) and Western blot. Dual luciferase target assay was used to verify the targeting relationship between miR-199a and HIF-1α gene.Results:The mRNA expression level of miR-199a in the HG group was significantly lower than that in the LG group and the HM group (all P<0.01), and the mRNA and protein expression levels of HIF-1α, FN and α-SMA in Hg group were significantly higher than those in LG group and HM group (all P<0.01). The mRNA and protein expression levels of HIF-1α, FN and α-SMA in the miR-199ai group were significantly higher than those in the miR-iNC group and the Mock group (all P<0.01). Target gene prediction by TargetScan and miRanda showed that miR-199a could target the 3′-untranslated region(3′-UTR) that binds HIF-1α. Dual luciferase reported results: Inhibition of miR-199a resulted in a significant increase in luciferase activity ( P<0.01), but there was no significant change in luciferase activity after 3 ′-UTR mutation of HIF-1α ( P>0.05). After transfection with si-HIF-1α, the mRNA and protein expression levels of HIF-1α, FN and α-SMA in si-HIF-1α group were significantly lower than those in NC group (all P<0.05). After transfection with miR-199ai, the mRNA and protein expression levels of HIF-1α, FN and α-SMA in miR-199ai group were significantly higher than those in NC group (all P<0.05). However, when co-transfected with si-HIF-1α and miR-199ai, the expression levels of mRNA and protein in miR-199AI group were significantly higher than those in NC group (all P<0.05). The mRNA and protein expression changes of HIF-1α, FN and α-SMA induced by transfection of si-HIF-1α or miR-199ai alone can be recovered. Conclusions:miR-199a can alleviate hyperglycemia-induced EMT in human renal tubular epithelial cells through targeted regulation of HIF-1α.

Epigenomic imbalance drives abnormal transcriptional processes,promoting the onset and progression of cancer.Although defective gene regulation generally affects carcinogenesis and tumor suppression networks,tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes,which may have significant implications for the development and application of epigenetic therapy,cancer immunotherapy,and their combinations.Herein,we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes,DNA methylation,histone post-translational modification,and chromatin structure in tumor immunogenicity,and introduce these epigenetic research methods.We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immuno-therapy through the complex interaction between cancer epigenetics and cancer immunology.