ObjectiveTo establish a new model of biliary peffusion in vitro,and to explore the feasibility of comparative biliary studies with the model.MethodsA set of ten-text-area model of biliary peffusion was designed in vitro.Saline,glycerol and pig bile were put into this model.The outflow of the first tube was set at 8 drops/min,and these liquids were collected through the ten sample test areas and measured at 0.5 hour,1.0 hour,1.5 hours,2.0 hours,12.0 hours and 24.0 hours.Equality of liquid amount was evaluated from each sample test area.Rods containing delayed release EDTA of ten different levels were placed in the test areas.The liquid was collected and EDTA concentrations were evaluated from the ten tubes at 24.0 hours.EDTA concentrations of the same rod in the three liquids were compared.ResultsA(1) the same time point,the amounts of saline,glycerol,pig bile flowing through the ten sample test areas showed no significant difference (P ＞ 0.01 ) ; but EDTA concentrations of the same liquid at 24.0 hours were significantly different (P ＜ 0.01 ) ; and EDTA concentrations of the same rod in the three liquids were also obviously different ( P ＜ 0.01 ).ConclusionThe same liquid flow rates and the same experimental environment can be achieved.Different test results can be obtained from different test areas in different experiments.The test results of the same kind of sample change with the changes of experimental conditions,which simulates biliary duct.

Pancreatic cancer is characterized by high invasiveness,high malignancy and poor prognosis. Therefore,it is urgent to find biomarkers for early diagnosis and to develop targeting drugs for treatment of pancreatic cancer. MicroRNAs (miRNAs)are single-stranded,non-coding small RNA molecules with a length of 18-25 nucleotides and are highly expressed in pancreatic cancer. MiRNAs play important roles in cell proliferation,differentiation,apoptosis,invasion and metastasis,and are associated with adverse prognosis and resistance to chemotherapy in pancreatic cancer. As a new target for diagnosis and treatment of pancreatic cancer,miRNA-21 has become the hot spot of clinical research. This article reviewed the advances in studies on miRNA-21 in the pathogenesis,diagnosis and treatment of pancreatic cancer.

In order to investigate the strength, structure and cell cytocompatibility of injectable thermosensitive chitosan (CS)/poly(vinyl alcohol) (PVA) composite hydrogel, chitosan hydrochloride solution was transferred to a neutral pH and mixed with different proportions of PVA, then the gelation time and strength of these different hydrogels were tested and spatial structures were observed under a scanning electron microscopy (SEM) after freeze-drying. The cytocompatibility of the hydrogels was evaluated through cytotoxicity test and three-dimensional culture with bone marrow mesenchymal stem cells. The results showed that the CS/PVA solution kept in liquid state at low temperature (0-4 degrees C) and turned into transparent elastomer about 15-20 min at 37 degrees C. Gelation time was prolonged, the strength increased and porous structure became dense with the PVA content increased in the mixed hydrogel. The cytotoxicity grades of these gels were from 0 to 1. Rabbit bone marrow mesenchymal stem cells could survive and proliferate in the gel within 3 weeks, and the gel had good cytocompatibility. It was concluded that thermosensitive CS/PVA composite hydrogel not only has interpenetrating network structure and better mechanical strength, but also has good cytocompatibility, and may be used as an injectable scaffold for tissue engineering.

Objective To evaluate the therapeutic effect of endoscopy for patients with anastomotic or nonanastomotic biliary strictures after liver transplantation.Methods Clinical and follow-up data of 56 patients,38 (67.9％) with anastomotic strictures (AS),and 18 (32.1％) with nonanastomotic strictures (NAS) after liver transplantation,who underwent endoscopic therapy were collected and analyzed.Results Compared to AS patients,NAS patients showed shorter time from liver transplantation to initial presentation (4.45 ± 1.47 vs.8.00 ±2.31 months,P =0.000),underwent more ERCP procedures (6.20 ±1.28 vs.4.11 ± 1.51,P =0.000) and received more stents (10.20 ±3.59 vs.7.53 ±2.12,P =0.001).Although the success rate was not statistically different,patients with NAS needed longer treatment duration and had higher recurrence rate (50.0％ vs.18.2％,P =0.033) as well as shorter duration from treatment to recurrence.Among patients with AS,the treatment failure group had longer stricture length (13.00 ±3.61 vs.6.63 ±2.09,P =0.000) than the success group and the patients who experienced recurrence had longer stricture length (10.48 ± 1.07 vs.5.86 ± 1.55,P =0.000) and narrower stricture diameter (1.52 ± 0.38 vs.1.94 ± 0.32,P =0.001).Among NAS patients,the multiple biliary strictures group had lower success rate and higher recurrence rate,but with no statistical significance (62.5％ vs.90.0％,P =0.275 and 66.7％ vs.37.5％,P =0.592,recpectively).Conclusion NAS patients have lower response to endoscopic treatment than AS patients.The length and diameter of biliary stricture in AS as well as the number of stricture in NAS may influent the effect of endoscopic treatment.

Objective To investigate the expression of the ternary complex factor Net in human pancreatic carcinoma cell line BxPC3 and its effect on cell proliferation and the expression of c-fos.Methods pEGFP-Net prokaryotic expression plasmid and empty vector pEGFP were transfed into BxPC3 cens by using lipofectamine 2000,then monoclonal cell which stably expressing Net was established.Human pancreatic carcinoma cells proliferation was detected by MTT and flow cytometry.The tuRNA and protein expression of Net and c-fos in BxPC3 cells were detected by real.time PCR and Western blot.Results Net was low expressed in BxPC3 cells.After pEGFP-Net transfection,Net wag stably expressed and the expression of c-fos was inhibited,cell proliferation was also inhibited after pEGFP-Net transfection,the inhibitory rates at the 3rd, 5th,7th day was 38.81%,55.34%and 56.92%respectively,which were significantly higher than those in the empty vector group(5.09%,12.42%,8.6%,P<0.05).G_0/G_1 phase cell was(61.79±5.67)%,which were significantly higher than(45.14±3.37)%in the empty vector group(P<0.05).Conclusions The ternary complex factor Net could inhibit pancreatic carcinoma cell line BxPC3 proliferation.Its mechanism was possibly repressing expression of oncogene c-fos.

Objective To observe the change of invasion and migration of the pancreatic carcinoma cell line SW1990 transfeeted with EEF1A2 gene.Methods Pancreatic carcinoma cell line SW1990 was transfected with EEF1A2 by recombinant adenovirus vector.The alteration of motility、invasion and adhesion property of SW1990 was evaluated by wound healing assay,transwell With or without Matrigel basement membrane and adhesion assay.Results Wound healing assay revealed that EEF1A2 enhanced cell motility and transwell assay with Matrigel indicated that the average numbers of transwell cells with EEFlA2 was increased from 23.25±5.23 to 65.42±8.24(P＜0.05).The adhesive rate was substantially increased in EEF1A2 transfected SW1990 cells compared with control cells.Conclusions EEF1A2 gene can promote the migration.invasion and adhesion ability of pancreatic cancer cell in vitro.It is indicated that EEF1A2 may involve in the development of human pancreatic cancer by influencing cell biological characteristics.

Objective To investigate the expressions of miR-21 in carcinoma tissue and serum of the patients with papillary thyroid carcinoma (PTC), and to clarify its clinical significance.Methods Real-time PCR was used to detect the expression levels of miR-21 in carcinoma tissue and serum of 46 PTC patients,thyroid tissue and serum of 35 cases of nodular goiter and 18 healthy people. The correlations of miR-21 expression level with the clinicopathological features of PTC patients were analyzed.Results The relative expression levels of miR-21 in carcinoma tissue and serum of the PTC patients were 2.74±1.53 and 0.21±0.08,and they were 0.92±0.81 and 0.08±0.05 in carcinoma tissue and serum of the patients with nodular goiter,0.86 ± 0.39 and 0.07 ± 0.03 in normal papillary thyroid tissue and serum of 18 healthy people.The expression levels of miR-21 in carcinoma tissue and serum of the patients with PTC were higher than those of the patients with nodular goiter and healthy people (P<0.05).The expression level of miR-21 in serum and carcinoma tissue of PTC patients had a positive correlation with the extraglandular invasion,distant metastasis,lymph node metastasis,and TNM stages (P<0.05);and it had no correlation with gender and tumor sizes.Conclusion The expression levels of miR-21 in carcinoma tissue and serum of the PTC patients are increased. The up-regulated expression of serum miR-2 1 is correlated with the clinicopathological features of PTC patients, which indicates that miR-2 1 could be used as a potential marker for early detection of PTC.

Objective To summarize the clinical experience of treating long bone defect with vascularized fibular graft.Methods From January,2008 to January,2015,31 cases of long bone defect were treated with vascularized fibula composite or not composite tissue flap graft.The length of transplanted fibula was 9-20 cm,and the flap area was 5 cm×3 cm to 21 cm×14 cm.All patients were followed up regularly.Limb function was assessed 12 months after surgery.Results Thirty-one cases of vascularized fibular flap survived after surgery.Thirty patients were followed up for 1.5 to 6 years (average,2.5 years).One patient was lost to follow-up.The bone defects of patients followed up were healed.There was one case fracture occurred for trauma,was treated with plaster cast for 6 months and healed.The transplanted fibular thickened for 1.3 to 2.5 years(average,1.6 years).Conclusion Vascularized fibular graft can reconstruct long bone defect for single use and shorten the duration of treatment with a good limb function.For cases combined soft tissue defect,vascularized fibula composite tissue flap can be applied to repair at the meantime.

In order to investigate the strength,structure and cell cytocompatibility of injectable thermosensitive chitosan(CS)/poly(vinyl alcohol)(PVA)composite hydrogel,chitosan hydrochloride solution was transferred to a neutral pH and mixed with different proportions of PVA,then the gelation time and strength of these different hydrogels were tested and spatial structures were observed under a scanning electron microscopy(SEM)after freeze-drying.The cytocompatibility of the hydrogels was evaluated through cytotoxicity test and three-dimensional culture with bone marrow mesenchymal stem cells.The results showed that the CS/PVA solution kept in liquid state at low temperature(0-4℃)and turned into transparent elastomer about 15-20 min at 37℃.Gelation time was prolonged,the strength increased and porous structure became dense with the PVA content increased in the mixed hydrogel.The cytotoxicity grades of these gels were from 0 to 1.Rabbit bone marrow mesenchymal stem cells could survive and proliferate in the gel within 3 weeks,and the gel had good cytocompatibility,it was concluded that thermosensitive CS/PVA composite hydrogel not only has interpenetrating network structure and better mechanical strength,but also has good cytocompatibility,and may be used as an injectable scaffold for tissue engineering.

Objective: To compare the safety and efficacy of the retrograde and the antegrade techniques in laparoscopic left hemihepatectomy. Methods: Of the 65 patients who underwent laparoscopic left hemihepatectomy between January 2016 to June 2018 at the Ningbo Li Huili Hospital of Medical Center, retrograde left hemihepatectomy was carried out in 31 patients, and antegrade left hemihepatectomy in 34 patients. The perioperative data, duration of operation, intraoperative blood loss, postoperative complications (including major bleeding, abdominal abscess and bile leakage), and post-operative hospital stay were retrospectively compared between the two groups. Results: There were no significant differences in the perioperative general status between the two groups (P>0.05). Both the duration of operation [(121.5±22.1) min vs. (190.9±48.9) min, P<0.05] and the amount of blood loss [(118.9±84.1) ml vs. (195.2±85.4) ml, P<0.05] were significantly less in the retrograde than the antegrade group. There were no statistical differences in the incidences of major complications such as post-hepatectomy hemorrhage, abdominal abscess, or bile leakage, and in the postoperative hospital stay [(10.0±2.8) d vs. (12.2±3.4) d, P>0.05]. Conclusions Occlusion of hepatic vascular inflow and outflow combined with retrograde left hemihepatectomy was safe and feasibly. The retrograde approach was superior to the antegrade approach in laparoscopic left hemihepatectomy.