Objective To compare the influence degree of laparoscopic operation and open operation on peripheral blood T lymphocyte subsets and Th1Th2 in patients with severe acute pancreatitis.Methods 54 patients who underwent surgical treatment in our hospital from February 2013 to December 2014 were selected as the subjects, 27 patients with severe acute pancreatitis who were treated with open operation as group A , 27 cases who were treated with laparoscopic operation at the same time were selected as group B , and then the peripheral blood T lymphocyte subsets and serum Th1Th2 indexes of two groups before the operation and at first,third and seventh day after the operation were respectively detected, then the detection results of two groups were compared.Results The peripheral blood T lymphocyte subsets and serum Th1Th2 indexes of group B at first, third and seventh day after the operation were all significantly better than those of group A , the difference was statistically significant ( P<0.05 ) , and the detection results of group A and group B at third day after the operation were worse than those at other time , the difference was statistically significant (P<0.05).Conclusion The influence of laparoscopic operation and open operation on T lymphocyte subsets and Th1Th2 in peripheral blood of patients with severe acute pancreatitis, the adverse effects of laparoscopic operation on the above indicators were relatively small.

Objective To study the antitumor effectivity of special cytotoxic T lymphocytes(CTLs) induced by B lymphocytes in mice.Methods B lymphocytes were collected,isolated and purfied.Cells were initially activated by CD40L and rmIL-4,then cocultivated with T lymphocytes.T lymphocyte proliferation was examined.Total RNA,which was extracted from Hepal-6(a hepatocelluar carcinoma cell line),were transfected into B lymphocytes,as experimental group;while transfected with RNA of mice liver cells,liposimes and 1640 were as control groups,The expression of antigen presenting cell(APC) markers(CD40,CD80 and CD86) and major histocomability complex(MHC) on B cell surface after transfection were deteced.CTL were obtained by stimulating T lymphocytes with transfected B lymphocytes.Hepal-6 was cell-targeted and examined as index of CTL killing activity.The IFN-r secretion of stimulated CTL was quantified.Results T cell proliferation in experimental group had a higher degree than that in RNA control group(P

Objective:To investigate the clinical efficacy of bilateral route minimal- incision necrosectomy combined with continuous lavage for the treatment of infected necrotizing pancreatitis (INP).Methods:The retrospective and descriptive study was conducted. The clinical data of 20 patients with IPN who were admitted to Daping Hospital, Army Medical University from April 2016 to July 2019 were collected. There were 11 males and 9 females, aged (42±9)years. All the 20 patients underwent bilateral route minimal-incision necrosectomy, and then be continuous perfused and drainage within the purulent cavity postoperatively. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up using outpatient examination and telephone interview was performed to detected patients fever, abdominal pain, abdominal distension, diarrhea, peripancreatic residual infection and survival up to January 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers. Results:(1) Surgical situations: of the 20 patients who underwent surgery successfully, 14 patients underwent upper abdomen combined with left retroperitoneal approach, 1 patient underwent upper abdomen combined with right retroperitoneal approach, and the other 5 patients underwent upper abdomen combined with bilateral retroperitoneal approach. Fourteen of the 20 patients underwent additional surgery including 10 cases undergoing jejunostomy, 2 cases undergoing gastrostomy combined with jejunostomy, 1 case undergoing laparoscopic cholecystectomy combined with jejunostomy, and 1 case undergoing cholecystectomy. The operation time and volume of intraoperative blood loss of 20 patients were (228±41) minutes and 100 mL (range, 50-700 mL), respectively. (2) Postoperative situations: 20 patients began continuous perfused with 0.9% sodium chloride solution within the purulent cavity at postoperative day 2 (range, day 1-14). Six of the 20 patients had postoperative complications including 1 case with postoperative gastric fistula combined with intraperitoneal hemorrhage who underwent laparotomy hemostasis combined with gastrostomy at day 13 postoperatively, 1 case with postoperative duodenal fistula who underwent gastrointestinal anastomosis and jejunostomy at day 111 postoperatively, 1 case with postoperative retroperitoneal residual tissue necrosis and infection who underwent peripancreatic necrotic tissue debridement and drainage at day 11 postoperatively, 1 case with postoperative gallbladder fistula who underwent cholecystectomy at day 71 postoperatively, and 2 cases with postoperative pancreatic fistula who were cured with conservative treatment. The duration of hospital stay after 1st operation of the 20 patients were 42 days (range,20-178 days). (3) Follow-up: all 20 patients were followed up for 6.0 to 45.0 months, with a median follow-up time of 14.5 months. During the follow-up, 1 case developed secondary diabetes, and none of patient showed clinical manifestation such as fever, abdominal pain, abdominal distension and diarrhea. The peripancreatic residual tissue of all 20 patients absorbed well, and none of patient died.Conclusion:Bilateral route minimal-incision necrosectomy combined with continuous lavage is safe and feasible for the treatment of INP.

AIM: To compare the methods of two currently employed isolation methods for endothelial progenitor cells (EPCs): from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133~+ cells, by defining the cell morphology, phenotype, reproductive activities and function in vitro, providing a reference for clinic application. METHODS: PBMCs from the healthy subjects were used for CD133~+ sorting or not. The two groups of isolated cells were suspended in complete medium M199 for 7 d to 14 d. EPCs phenotype were characterized by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and VEGF concentration was measured using an ELISA kit. Matrigel experiment and migration assay were imitated vascularization in vivo. RESULTS: PBMCs produced more colony-forming units (CFU) than CD133~+ cells from the same volume of blood (P<0.01). From 7 d to 14 d, the two groups show decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144~+ cells in CD133~+ group were lower than those in PBMCs groups (P<0.01). Cells in PBMCs group secreted more VEGF than that in CD133~+ group on 7 d (P<0.01). Compared to CD133~+ group, PBMCs group showed more potential of proliferation and vascularization in vitro. CONCLUSION: CD133~+ sorted cells show a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, which is unable to differentiate to mature endothelial cells, indicating that it's not a preferential way to obtain EPCs for clinic therapy.

Two isolation methods for sorting of endothelial progenitor cells (EPCs): from peripheral blood mononuclear cells (PBMCs) and CD133(+) enriched cells were compared, by defining the cell morphology, phenotype, reproductive activities and function in vitro, to provide a reference for clinical application of EPCs. PBMCs from healthy subjects were used either directly for cell culture or for CD133(+) sorting. The two groups of cells were cultured in complete medium 199 (M199) for 7 to 14 days and the phenotypes of EPCs were analyzed by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and the VEGF concentration was measured using an ELISA kit. ECM gel experiment and migration assay were performed in vivo. The results showed that PBMCs produced more colony-forming units (CFU) than CD133(+) enriched cells from the same volume of blood (P<0.01). From day 7 to 14, the two groups showed decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144(+) cells in CD133(+) group were less than in PBMCs group (P<0.01). PBMCs group secreted more VEGF than CD133(+) group on the day 7 (P<0.01). As compared with CD133(+) group, PBMCs group had more potent potential of proliferation and vascularization in vitro. It was concluded that CD133(+) sorted cells showed a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, suggesting that CD133-negative cells may be a preferential way to get EPCs for clinical therapy.

Objective: To investigate the expression of long noncoding RNA (lncRNA) LINC01296 in hepatocellular carcinoma (HCC) tissues and cells, and to explore its effects on the proliferation, migration and invasion of Hep3B cells. Methods: The expression of LINC01296 in liver cancer cells, HCC tissues and para-cancerous tissues was analyzed using Gene Expression Profiling Interactive Analysis (GEPIA) database and real-time quantitative PCR. Moreover, the expression of LINC01296 in HCC cells and the normal hepatocyte cells was detected by real-time fluorescent quantitative PCR. After the lentivirus carrying LINC01296-overexpression or-silencing vector was infected into Hep3B cells, the cell proliferation, migration and invasion were detected by CCK-8 assay and Transwell chamber assay, respectively. The effect of LINC01296 on the expressions of epithelial-mesenchymal transition (EMT)-related proteins in Hep3B cells was detected by Western blotting. Results: The expression level of LINC01296 was significantly up-regulated in HCC tissues and cells (both P < 0.05). The overexpression of LINC01296 significantly promoted the proliferation, migration, invasion and EMT activities of Hep3B cells (all P < 0.01), whereas the results were opposite when LINC01296 was silenced (all P < 0.01). Conclusion: The expression level of LINC01296 is up-regulated in HCC tissues and cells, and it can promote the proliferation, migration and invasion of Hep3B cells by enhancing the activities of EMT.

Objective To study the effect of combined laparoscopic and percutaneous nephroscopic necrosectomy in the treatment of peripancreatic abscesses.Methods The clinical data of 8 patients with peripancreatic abscesses treated by combined laparoscopic and percutaneous nephroscopic necrosectomy in our hospital from June 2015 to June 2017 were retrospectively analyzed.Results 8 patients were treated with percutaneous puncture and drainage under Ultrasonic / Computed Tomography guidance.Combined laparoscopic and percutaneous nephroscopic necrosectomy was then carried out.Two patients underwent percutaneous nephroscopic surgery twice and laparoscopic surgery once,and 3 patients underwent percutaneous nephroscopic surgery thrice,and 3 patients underwent percutaneous nephroscopic surgery 4 times and laparoscopic surgery once.One patient after percutaneous nephroscopic necrosectomy was complicated with sinus tract hemorrhage,which was treated by haemostasis through a small incision.Two patients who developed postoperative colonic fistula were treated successfully by conservative treatment.The average length of hospital stay was 80 d (60 ～ 153 d),and there was no death.Conclusion Combined laparoscopic and percutaneous nephroscopic necrosectomy was a minimally invasive and efficacious method to treat peripancreatic abscesses.

Two isolation methods for sorting of endothelial progenitor cells(EPCs): from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive activities and function in vitro,to provide a reference for clinical application of EPCs.PBMCs from healthy subjects were used either directly for cell culture or for CD133+ sorting.The two groups of cells were cultured in complete medium 199(M199)for 7 to 14 days and the phenotypes of EPCs were analyzed by FACS.The proliferation of differentiated EPCs was studied by MTT assay,and the VEGF concentration was measured using an ELISA kit.ECM gel experiment and migration assay were performed in vivo.The results showed that PBMCs produced more colony-forming units(CFU)than CD133+enriched cells from the same volume of blood(P<0.01).From day 7 to 14,the two groups showed decreased expression of hematopoietic stem cell markers and increased level of endothelial markers,but CD144+cells in CD133+ group were less than in PBMCs group(P<0.01).PBMCs group secreted more VEGF than CD133+group on the day 7(P<0.01).As compared with CD133+ group,PBMCs group had more potent potential of proliferation and vascularization in vitro.It was concluded that CD133+sorted cells showed a lower capacity of differentiation,secretion,proliferation and vascularization in vitro,suggesting that CD133-negative cells may be a preferential way to get EPCs for clinical therapy.

OBJECTIVECellular senescence as one of the important steps against tumor is observed in many cancer patients receiving chemotherapy and is related to chemotherapeutic response. To investigate the effect of cisplatin on hepatocellular carcinoma, we treated HepG2 cells exhibiting wild-type TP53 with gradient concentrations of cisplatin.

METHODSThe inhibitory effects of cisplatin on human hepatoma HepG2 cells were detected by MTT assay and colony formation test. The changes in cell cycle were analyzed by flow cytometry, and cellular senescence was detected with senescence associated β-galactosidase (SA β-gal) staining. The relative mRNA expression levels of TP53, P21 and P19 was estimated using semi-quantitative real-time RT-PCR, and the protein expressions of P53 and P21 were detected using Western blotting.

RESULTSCisplatin induced irreversible proliferation inhibition and G1 phase arrest of HepG2 cells. Elevated levels of senescence-associated β-galactosidase was observed in HepG2 cells exposed to low doses of cisplatin. P19 expression immediately increased following cisplatin exposure and reached the maximum level at 48 h, followed then by a rapid decrease to the baseline level, whereas the expressions levels of TP53 and P21 mRNA increased continuously. Western blotting confirmed P53 and P21 expression changes similar to their mRNA expressions during cisplatin-induced cellular senescence in HepG2 cells.

CONCLUSIONOur results revealed a functional link between cisplatin and hepatocellular senescence. Cellular senescence induced by cisplatin as a stabile senescent cellular model can be used for further research.

Cell Cycle ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cellular Senescence ; Cisplatin ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p19 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Hep G2 Cells ; Humans ; Tumor Suppressor Protein p53 ; metabolism ; Up-Regulation

BACKGROUNDAccumulating evidence indicates that systemic inflammation response is associated with the prognosis of various cancers. The aim of this study was to investigate the neutrophil-lymphocyte ratio (NLR), which is one of the systemic inflammation markers, in the prognosis of hepatocellular carcinoma (HCC) after treatment of transcatheter arterial chemoembolization (TACE).

METHODSThe clinical data of 178 HCC patients who received TACE were retrospectively analyzed. The optimal NLR cutoff was determined according to the receiver operating characteristic (ROC) analysis. All patients were divided into NLR-normal group and NLR-elevated group according to the cutoff, and the clinical features of these two groups were comparatively analyzed. Meanwhile, the overall survival and disease free survival (DFS) were analyzed using the Kaplan-Meier method. The risk factors of postoperative survival were investigated using univariate and multivariate Cox regression analyses.

RESULTSThe optimal NLR cutoff was defined at 1.85 and 42 (23.6%) patients had an elevated NLR (NLR>1.85). The median survival time was 9.5 months (range 1-99 months). The clinical data between the two groups were comparable, except for a-fetoprotein. Follow-up results showed that the median survival of patients with normal NLR was 17.5 months (range: 1-99 months) compared with 8 months (range: 8-68 months) of patients with elevated NLR. The 1, 3 and 5-year overall survival of patients in the NLR-normal group and NLR-elevated group were 57.3%, 44.1%, and 27.2% and 42.1%, 19.6%, and 9.5% respectively (χ(2) = 194.2, P < 0.001). Similarly, the disease free survival also has a significant difference (χ(2) = 39.3, P < 0.001). Multivariate Cox regression analysis showed that a high NLR was an independent factor affecting the survival rate of HCC after TACE (P = 0.04).

CONCLUSIONPreoperative NLR was an important prognostic factor to predict the prognosis of patients with intermediate HCC treated with TACE.

Adult ; Aged ; Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Lymphocytes ; metabolism ; physiology ; Male ; Middle Aged ; Neutrophils ; metabolism ; physiology