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OB JECTIVE To study the protective effects of saponins from Gleditsia sinensis on ischemic stroke with phlegm and blood stasis （ISPBS）model rats ，and to explore its mechanism. METHODS Totally 119 rats were randomly divided into normal group （normal saline ），sham operation group （normal saline ），model group （normal saline ），nimodipine group （positive control group ，5 mg/kg），G. sinensis saponin low-dose ，medium-dose and high-dose groups （3.21，6.42 and 12.84 mg/kg），with 17 rats in each group. Except for normal group ，other groups were all given high-fat diet+suture-occluded method to induce ISPBS model. The neurological function score ，water content of brain tissue ，pathological morphology of brain tissue ，the changes of hemorheology indexes （whole blood viscosity ， erythrocyte aggregation index ， Casson-viscosity）， four items of blood lipid [triacylglycerol （TG），total cholesterol （TC），low-density lipoprotein cholesterol （LDL-C），high-density lipoprotein cholesterol（HDL-C）] and inflammatory factors in serum and oxidative stress indexes [malondialdehyde （MDA），nitric oxide （NO），superoxide dismutase （SOD）] in brain tissue were determined or observed in rats. The protein expressions of B lymphocytoma 2（Bcl-2），Bcl-2 associated X protein （Bax）and caspase- 3 in cerebral tissue were also detected. RESULTS Compared with normal group ，the score of nerve function ，5 kinds of serum indexes （TC，TG，LDL-C，TNF-α，IL-1β）， hemorheology indexes ，the contents of MDA and NO and protein expressions of Bax and caspase- 3 in cerebral tissue were all increased significantly in model group （P＜0.01）. The levels of HDL-C and IL- 10 in serum ，SOD activity and protein expression of Bcl- 2 in cerebral tissue were decreased significantly （P＜0.01），and obvious lesions such as nuclear pyknosis and cell membrane fragmentation occurred in brain tissue. Compared with model group ，above indexes of administration groups were improved to different extents ，among which there was statistical significance in above indexes of G. sinensis saponin high-dose group （P＜ 0.01）. CONCLUSIONS Saponin from G. sinensis has a good protective effect on ISPBS model rats. Its mechanism may beassociated with reducing oxidative damage ， reducing the production of pro-inflammatory mediators and resisting neuronal apoptosis.
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Objective:To investigate the effects of long non-coding RNA (lncRNA) FBXL19-AS1 on the proliferation, migration and invasion of pancreatic cancer cells, and to determine the targeting relationship of lncRNA FBXL19-AS1 and microRNA-339-3p (miR-339-3p).Methods:From January 2017 to August 2019, 73 cancer tissues and matched normal pancreatic tissues adjacent to cancer from patients pathologically diagnosed as pancreatic cancer who underwent surgical resection in Yantai Hospital of Yantai were collected. Normal pancreatic epithelial cells (hTERT-HPNE) and 3 pancreatic cancer cell lines (Capan-1, SW1990, PaTu8988) were cultured in vitro. The real-time fluorescent quantitative PCR was used to detect the expression of lncRNA FBXL19-AS1 and miR-339-3p in pancreatic cancer tissues and cell lines. The Capan-1 cells were divided into the NC group (normal control group), si-NC group (transfected with meaningless negative sequence), si-FBXL19-AS1 group (transfected with FBXL19-AS1 small interfering RNA), miR-NC group (transfected with empty plasmid control), miR-339-3p group (transfected with miR-339-3p overexpression lentiviral vector), si-FBXL19-AS1+ anti-miR-339-3p NC group (cotransfected with FBXL19-AS1siRNA and miR-339-3p inhibitor negative control sequence) and si-FBXL19-AS1+ anti-miR-339-3p group (cotransfected with FBXL19-AS1siRNA and miR-339-3p inhibitor). CCK-8 method was used to detect cell proliferation activity. Transwell chamber was used to detect cell migration and invasion ability, and western blotting method was used to detect cell cyclinD1, matrix metalloproteinase 2 (MMP2) and MMP9 protein expression. Bioinformatics and dual luciferase reporter gene experiments were used to analyze the targeting relationship between lncRNA FBXL19-AS1 and miR-339-3p.Results:The expression of lncRNA FBXL19-AS1 in pancreatic cancer tissue was significantly higher than that in normal pancreatic tissue adjacent to cancer (2.96±0.21 vs 1.00±0.13, P<0.05), and the expression of miR-339-3p was significantly lower than that in normal pancreatic tissue adjacent to cancer (0.37±0.05 vs 1.00±0.11, P<0.05). The expression of lncRNA FBXL19-AS1 in Capan-1, SW1990 and PaTu8988 cells were 2.43±0.18, 1.97±0.13 and 1.73±0.14, respectively, which were significantly higher than that of hTERT-HPNE cells 1.00±0.07. The expression of miR-339-3p were 0.42±0.03, 0.54±0.03 and 0.57±0.04, respectively, which were all significantly lower than 1.00±0.05 of hTERT-HPNE cells. Among them, the expression of lncRNA FBXL 19-AS1 in Capan-1 cells was the highest, and the miR-339-3p was the lowest. Compared with the si-NC group, the absorbance value ( A450) of Capan-1 cells in the si-FBXL19-AS1 group, the number of migrating cells, and the number of transmembrane cells were significantly decreased (0.47±0.03 vs 0.94±0.06, 81.00±7.41 vs 187.00±16.13, 67.00±5.41 vs 141.00±9.24), the protein expression of cyclinD1, MMP2 and MMP9 was significantly reduced (0.44±0.03 vs 0.83±0.05, 0.48±0.03 vs 0.92±0.07, 0.38±0.02 vs 0.73±0.05). Compared with the miR-NC group, the A450, the number of migrating cells, and the number of transmembrane cells of Capan-1 cells in the miR-339-3p group were significantly decreased (0.54±0.04 vs 0.94±0.05, 98.00±6.53 vs 193.00±10.02, 83.00±6.58 vs 146.00±7.11, the protein expression of cyclinD1, MMP2 and MMP9 was significantly reduced (0.47±0.03 vs 0.81±0.07, 0.43±0.03 vs 0.94±0.06, 0.32±0.02 vs 0.71±0.06). Compared with the si-FBXL19-AS1+ anti-miR-NC group, the A450, the number of migrating cells and the number of transmembrane cells in the si-FBXL19-AS1+ anti-miR-339-3p group increased significantly (0.96±0.07 vs 0.48±0.03, 204.00±11.25 vs 83.00±5.11, 157.00±8.76 vs 64.00±4.12, P all <0.05), the protein expression of cyclinD1, MMP2 and MMP9 increased significantly (0.84±0.06 vs 0.42±0.03, 0.96±0.08 vs 0.47±0.08, 0.74±0.06 vs 0.37±0.02, P all <0.05). The luciferase activity of Capan-1 cells cotransfected with WT-FBXL19-AS1 and miR-339-3p mimics was significantly lower than that of the cotransfected with WT-FBXL19-AS1 and miR-NC (0.47±0.04 vs 1.00±0.10, P all <0.05). The difference of the luciferase of Capan-1 cells in the cotransfected MUT-FBXL19-AS1 and miR-339-3p mimics group and the cotransfected MUT-FBXL19-AS1 and miR-NC group was not statistically significant. Conclusions:LncRNA FBXL19-AS1 was highly expressed in pancreatic cancer tissues and Capan-1 pancreatic cancer cell lines. Knockdown of lncRNA FBXL19-AS1 can target miR-339-3p to regulate the proliferation, migration and invasion of pancreatic cancer cells, and promote the occurrence and development of pancreatic cancer.
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OBJECTIVE：To establish and optimize a extraction method of Fructus Gleditsiae Abnormallis ，and to analyze and identify chemical components of the extract simultaneously. METHODS ：Fructus Gleditsiae Abnormallis was extracted with CO 2 supercritical fluid extraction （SFE）method. Based on single factor tests ，using extraction yield as index ，extraction temperature ， extraction pressure and extraction time as investigation factors ，SFE technology was optimized with orthogonal test ，and validation test was performed. Chemical components in the extract were identified by GC-MS. Relative percentage of each component was calculated with area normalization method. RESULTS ：The optimal SFE extraction technology of Fructus Gleditsiae Abnormallis was extraction temperature of 60 ℃，extraction pressure of 300 MPa and pression time of 15 min. Average extraction of 3 times of validation tests was 1.73％（RSD＝1.78％，n＝3）. The 48 components in the extracts of Fructus Gleditsiae Abnormallis were identified，which accounted for 98.31％ of the total amount of the extracts. The extracts of Fructus Gleditisae Abnormalis mainly included organic acids ，accounting for 36.99％，followed by alkaloids ，accounting for 12.59％ in total. Main components were palmitic acid （16.62％），oleic acid （14.12％），N-aminotetrahydropyrrole（9.79％），2，6-dimethyloctane-1，7-dien-3-ol（5.95％）， tetrahydropyran（3.83％），vanillin（3.39％），etc. CONCLUSIONS ：SFE method of Fructus Gleditisae Abnormalis is established successfully，and the extract is mainly organic acids.
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ObjectiveTo investigate the treatment strategies and factors influencing the prognosis of patients with primary gallbladder carcinoma.MethodsThe clinical data of 135 patients with primary gallbladder cancer who were admitted to the Cancer Hospital of Tianjin Medical University from January 2000 to December 2009 were retrospectively analyzed.The survival curve was drawn by the Kaplan-Meier method,and the survival rates were analyzed by using the Log-rank test.Factors which may have influences on the prognosis were analyzed by univariate analysis and COX multivariate analysis.ResultsThe overall 1-,3-,5-year survival rates of the 135 patients were 46.7％,10.4％ and 5.2％,respectively.The 1-,3-,5-year survival rates of 74 patients who received radical resection of gallbladder cancer were 68.9％,18.9％ and 9.5％,respectively.The 1-,3-,5-year survival rates of 50 patients who received palliative treatment were 24.0％,0 and 0,respectively.The 1-,3-,5-year survival rates of 11 patients who received conservative treatment were 0,0 and 0,respectively.There was no significant difference in the survival rates among patients who received different treatment methods (x2 =5.642,P ＜ 0.05 ). Of the 9 patients with gallbladder cancer who received reoperation after laparoscopic choledochotomy,the survival time of 1 patient in stage Ⅰ and 1 of the 3 patients in stage Ⅱ who received radical surgery exceeded 5 years,while the survival time of 5 patients in stage Ⅱ who received palliative treatment was shorter than 5 years.There was a significant difference in the survival time among the 3 groups of patients ( x2 =5.642,P＜0.05).Under the condition of same TNM stages ( Ⅱ,ⅢA,ⅢB,ⅣA,ⅣB),the survival rates of patients who received radical resection of gallbladder cancer were significantly higher than those who received palliative or conservative treatment ( x2 =8.971,21.250,44.153,6.696,21.722,P ＜ 0.05 ).The results of univariate analysis showed that age,CA19-9,TNM stages and treatment methods were risk factors influencing the median survival time ( x2 =8.466,3.977,9.837,5.642,P ＜ 0.05 ).The results of multivariate analysis showed that age,TNM stages and treatment methods were the independent risk factors influencing the median survival time ( Wald=5.779,14.724,11.640,P＜0.05).ConclusionThe prognosis of primary gallbladder cancer is poor.Age,TNM stages and treatment methods are the independent factors influencing the prognosis of patients with gallbladder cancer,and patients who receive radical resection have relatively good prognosis.
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ObjectiveTo evaluate the surgical outcomes and prognosis of patients after liver resection for noncolorectal liver metastases.Methods72 patients with liver metastases who underwent liver resection at Tianjin Medical University Cancer Hospital were retrospectively studied.There were 32 males and 42 females,aged between 35～78 years.After liver resection,68 patients had a R0 resection (negative histological margin),and 4 patients had a R1 resection (positive histological margin).The primary tumours were breast,(n =16,22.2 ％),lung (n =14,19.4 ％),gastrointestinal (n=12,16.7％),gynecological (n =8,11.1％),pancreatobiliary (n =8,11.1％),melanoma (n=4,5.6％),sarcoma (n=4,5.6％),and genitourinary (n=2,2.8％).The mean diameter of the main tumour was 4.8 cm (range,1.5- 11.0 cm).The mean number of liver metastases was 1.2 (range,1-5).Liver metastases were synchronous in 6 patients (8.3％) and metachronous in the remaining 66 patients (91.7％).ResultsThe operative mortality was 0％.The mean hospital stay was 14.4 days (range 6-67 days).The median overall survival was 31 months (range,6-127 months).The 1-,3- and 5-year survival rates were 81.9％,37.5％ and 23.6％,respectively.Univariate analysis revealed primary tumour sites (breast vs.others),histological type (adenocarcinoma vs.others),postoperative chemotherapy,number of liver metastases (solitary vs.multiple) and time to liver metastases from diagnosis of primary tumours (≤ 12 months vs.＞ 12 months) were associated with overall survival (all,P＜0.05).In multivariate analysis,factors independently associated with poor survival were nonbreast origin (P =0.012),time to liver metastases from diagnosis of primary tumours ＜12 months (P=0.027) and multiple liver metastases (P=0.008).ConclusionsIn selected patients,liver resection is an effective and safe treatment for noncolorectal liver metastases.The time to liver metastases from diagnosis of primary tumours was independently associated with overall survival.For solitary or liver metastasis of breast origin,surgical resection significantly improved survival.