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Objective: To observe the antitumor effect of triptolide in the human pancreatic cancer tumor-bearing nude mice, and to explore its molecular mechanism. Methods: The healthy BALB/ c nude mice were selected and transplanted with the human pancreatic cancer SW1990 cells into the skin of the right hind limb to establish the tumor-bearing model of nude mice. The successful modeling nude mice were divided into model group, gemcitabine group and triptolide group, 10 in each group; another 10 healthy nude mice were selected as control group. The tumor tissue of SW1990 pancreatic cancer tumor-bearing nude mice were weighed and the inhibitory rate of tumor was calculated. The expressions of apoptosis-related proteins Bax and bcl-2 in the pancreas tissue of the nude mice in each group were detected by rmmunohistochemistry. The expression levels of Bax, bcl-2, Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in each group were detected by Western blotting method. Results: Compared with gemcitabine group, the inhibitory rate of tumor of SW1990 pancreatic cancer in triptolide group was significantly increased (P < 0. 05). The HE staining results showed that compared with model group, the proliferation of pancreatic cancer tumor cells in triptolide group was inhibited. The immunohistochemistry results showed that the expression level of Bcl-2 in the pancreas tissue and the Bcl-2 /Bax ratio of the nude mice in tripptolide group were lower than those in model group (P < 0 . 05). The Western blotting results showed that compared with model group, the expression levels of Bax, Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in triptolide group were significantly increased (P < 0 . 05), while the expression level of Bcl-2 was significantly decreased (P < 0 . 05). Conclusion: Triptolide can inhibit the growth of tumor tissue in the nude mice; its mechanism may be related to inhibiting the activation of Bcl-2 in pancreatic tumor tissue, promoting the expression of Bax, reducing the ratio of Bcl-2 /Bax, promoting the activation of Caspase-9 and Caspase-3, and inducing the apoptosis of pancreatic cancer transplanted tumor cells.
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Objective To investigate the expression of miRNA-182 in hepatocellular carcinoma (HCC) cell lines and tissues, to verify the targeted regulation relationship between miRNA -182 and forkhead box transcription factor 2 (FOXF2) and to explore whether miRNA-182 can promote the invasion, metastasis and angiogenesis of HCC by targeting FOXF2.Methods From January to October 2017, a total of 41 patients with primary HCC admitting to Cancer Hospital Affiliated to Zhengzhou University and receiving operation were enrolled.The expression of miRNA-182 in tumor tissues and normal paracancerous tissues was detected by quantitative real-time fluorescence polymerase chain reaction ( qRT-PCR).The effects of overexpression of miRNR-182 and FOXF2 on invasion and metastasis of HCC were detected by Transwell transfer experiment and the Boyden invasion experiment.The effects of miRNA-182 and FOXF2 on angiogenesis of HCC were determinded by in vitro angiogenesis experiments.The targeted regulation relationship between miRNA -182 and FOXF2 was investigated by dual luciferase reporter gene .The expression of FOXF2 at mRNA and protein level after miRNA-182 and miRNA-182 scramble transfection was detected by qRT -PCR and Western blotting method.T test was performed for statistical analysis.Results The expression in miRNA-182 in HCC tissue was higher than that in normal tissues adjacent to cancer (5.41 ±1.72 vs.1.80 ±0.76), and the difference was statistically significant (t =-5.764, P <0.01).The results of Transwell transfer experiment and the Boyden invasion experiment showed that the number of cells passing through the membrane of the small chamber of miRNA -182 over-expression group was higher than that of miRNA-182 scramble group (85.65 ±4.86 vs.31.43 ±2.41, 55.34 ±5.66 vs. 19.13 ±2.12), the number of cells passing through the membrane of the small chamber of miRNA -182 +FOXF2 over expression group was less than that of simple miRNA -182 over expression group (11.21 ±3.16 vs. 31.43 ±2.41, 8.67 ±2.83 vs.19.13 ±2.12), and the differences were statistically significant (t =15.110, 9.220, 5.443 and 4.410; all P <0.05).The activity of luciferase of wild FOXF2 and miRNA-182 cotransfection group was lower than that of miRNA -182 scramble group (0.43 ±0.10 vs.0.97 ±0.05), and the difference was statistically significant (t =8.042, P =0.012).However the activity of luciferase of mutated FOXF2 and miRNA-182 cotransfection group was higher than that of miRNA -182 scramble group (0.97 ±0.47 vs.1.06 ±0.52), and the difference was not statistically significant (t =0.934, P =0.402).The results of in vitro angiogenesis experiments demonstrated that the number of constituent vessels in miRNA -182 over expression group was higher than that in miRNA-182 scramble group (14.27 ±1.77 vs.5.91 ±1.42), and after FOXF2 over-expressing plasmids cotransfected, the angiogenesis ability restored (1.78 ±0.71 vs.14.27 ±1.77), and the differences were statistically significant (t =6.530 and 4.570, both P <0.05).The results of Western blotting indicated that the expression level of FOXF 2 in miRNA-182 transfection group was lower than that of miRNA-182 scramble group (1.01 ±0.13 vs.4.32 ±0.46), and the difference was statistically significant (t =7.420, P =0.012).The results of qRT-PCR showed that the expression level of FOXF2 mRNA of miRNA-182 transfection group was lower than that in miRNA-182 scramble group (0.591 ±0.284 vs.1.534 ±0.345), and the difference was statistically significant (t =3.465, P =0.014).Conclusion miRNA-182 can promote the invasion and metastasis of HCC cell line by targeting regulation of FOXF 2.
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Objective To select the best preoperative biliary drainage (PBD) method for patients with hilar cholangiocarcinoma.Methods The PubMed,EMBASE,Web of Science,CNKI,and Wanfang Database were systematically searched for prospective or retrospective studies on biliary drainage for patients with hilar cholangiocarcinoma with obstructive jaundice.The drainage-related cholangitis,pancreatitis,hemorrhage,and the success rate in relieving jaundice were analyzed.The meta-analysis was performed using the Review Manager 5.3 and the stata 12.0 using a fixed or random effects model.Results This meta-analysis included 12 studies with 1567 patients.The results showed a lower risk of cholangitis with PTBD than EBD (RR=0.60,95％CI:0.39～0.95,P＜0.05).PTBD also resulted in a lower risk of pancreatitis than EBD (RR=0.30,95％CI:0.15～0.59,P＜0.05),and a higher rate of successful relief of cholestatic jaundice (RR=2.77,95％CI:1.79～4.28,P＜0.05).However,the risk of bleeding for PTBD was higher (RR=2.38,95％ CI:1.12～5.05,P＜0.05),the risk of intraoperative blood transfusion increased (RR=1.59,95％ CI:1.05-2.42,P＜0.05),and the risk of celiac metastasis was also increased (PR=3.24,95％CI:1.15～9.12,P＜0.05) when compared with EBD.The incidence of celiac metastasis was as high as 4.2％.There were no significant differences between PTBD and EBD in the rates of bile leakage,intra-abdominal abscesses,hemorrhage,R0 resection,postoperative hospital stay postoperative complications and in-hospital mortality,what's more,there were no significant differences in the incidence of cholangitis,pancreatitis,and liver abscess between ENBD and EBS.Conclusions The postoperative hospital stay was similar between the two groups.ENBD was a better choice than PTBD for patients who required PBD.PTBD could be used after the failure of ENBD.
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Objective: To establish the pancreatic carcinoma models of rats with dimethybenzanthracene (DMBA)embedding method,to discuss the treatment effect of Da Huang Mu Dan Tang (DHMDT)in the rats with pancreatic carcinoma and the protective effects on the liver and kidney function,and to clarify the mechanisms. Methods:Sixty-five SD male rats were divided into blank control (n=10)and modeling group (n=55).DMBA was embedded in the capsule of pancreas to establish the pancreatic carcinoma models of rats. After 3 months, 50 successfully modeling rats were randomly divided into model,5-FU (intravennous injection,12 mg· kg-1 ), low,middle and high doses of DHMDT (intragastric administration,20,40 and 80 mg· kg-1 )groups.After 10 weeks of administration,the levels of alanine aminotransferase (ALT),aspartate transaminase (AST),urea nitrogen (BUN),creatinine (Cr)and total bilirubin (TBiL)in serum of the rats in various groups were measured;the weights and the inhibitory rates of the pancreatic tumor were detected;TUNEL method was used to analyze the apoptotic rates of tumor tissue;the expression levels of SST2R,Bax and Bcl-2 genes in pancreatic tumor tissue of the rats were detected by Q-PCR method.Results:Compared with model group,after treated for 10 weeks,the levels of ALT,AST,BUN,Cr and TBiL in serum of the rats in low,middle and high doses of DHMDT groups were significantly decreased (P <0.05 or P <0.01);the weights of tumor of the rats in middle and high doses of DHMDT groups were decreased (P <0.05 or P <0.01);the apoptotic rates of the cells in pancreatic tumor tissue of the rats in low,middle and high doses of DHMDT groups were increased (P <0.05 or P <0.01);the expression levels of SST2R gene in pancreatic tumor tissue of the rats in low,middle and high doses of DHMDT groups were significantly increased (P <0.05 or P <0.01);the expression levels of Bax gene in pancreatic tumor tissue of the rats in middle and high doses of DHMDT groups were significantly increased (P < 0.05 or P < 0.01),while the expression levels of Bcl-2 in pancreatic tumor tissue of the rats were significantly decreased (P < 0.05 ). Conclusion:DHMDT has the treatment effect in the rats with pancreatic carcinoma and can promote the apoptosis of tumor cells;DHMDT can protect the liver and kidney function of the rats with pancreatic carcinoma.
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Objective To investigate the anti-tumor efficiency of cytotoxic T-lymphocyte (CTL) induced by activated B lymphocyte after hepatocellular carcinoma (HCC) alpha fetoprotein (AFP) mRNA transfection.Methods B lymphocytes were fractionated,purified and activated by recombinant human soluble sCD40L.PGEM4Z/AFP/A64-EGFP plasmid was established in vitro,mixed with polymerase T7RNA,and then transcribed into AFP mRNA with Poly (A) sequence.B lymphocytes electrotransfected with AFP mRNA were in the experimental group,B lymphocytes electrotransfected with GAPDH mRNA were in the negative control group,and untreated B lymphocytes were in the blank control group.The expressions of antigen-presenting cell (APC)markers (CD19,CD20,CD21,CD40,CD80,CD83) and major histocompatibility complex (MHC) of the 3 groups were detected.B lymphocytes of the 3 groups were cultured with T lymphocytes at ratios of 1∶40,1 ∶ 20,1∶10 and 1∶5 to induce and ampify CTL,and then the absorbance values were detected to evaluate the proliferation ability of T lymphocytes.The killing activity of CTL was investigated with HCC cell line SMMC7721 as the target cells.All data were analyzed using the paired t test,one-way analysis of variance or Tamhane's T2 test.Results The expressions of CD19,CD20,CD21,CD40,CD80 and CD83 of the experimental group were 74 ± 11,78 ±8,80 ± 10,90 ± 11,82 ± 6,56 ± 5,which were significantly higher than 51 ± 5,60 ± 7,53 ± 5,73 ± 8,50 ± 5 and 49 ± 6 of the negative control group,and 46 ± 3,54 ± 5,41 ± 3,56 ± 5,52 ± 6 and 21 ± 4 of the blank control group (t =5.302,4.812,7.627,5.932,9.142,7.813; 11.581,7.036,13.592,12.873,9.235,14.619,P ＜ 0.01).The proliferation of CTL of the experimental group was significantly higher than that in the negative control group and blank control group (t =18.203,23.714,15.062,9.417 ; 16.833,19.392,13.871,6.592,P ＜0.01).When the T lymphocytes were mixed with the HCC cell line SMMC7721 at the ratios of 40∶ 1,20∶1and 10∶1,the killing rates of HCC cells by CTL of the exprimental group were 43％ 4％,32％ ± 4％ and 22％ ±3％,which were significantly higher than 15％ ± 5％,7％ ± 3％ and 6％ ±2％ of the negative control group,and 7％±3％,8％±3％ and 9％±4％ of the blank control group (t =9.141,13.272,11.901; 14.372,12.835,9.507,P ＜ 0.01).Conclusion Activated B lymphocytes after HCC AFP mRNA transfection may effectively induce CTL to kill HCC cells.
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To explore the relations of Morphology Immnuophe-notype Cytogenetics Molecular biology(MICM) detection on diagnosis andtreat,emt of leukemia． Methods: 68 cases of leukemia patients had beenanalyzed by morphology(FAB)． Immunohistochemistry(Flow Cytometry, FCM)． chromosome G banding technique and dual-color fluorescence insitu hybridization (D-FISH)．Technique:All patients were treated bychemotherapy． T test and X2 test of significance． Results: 7 cases have acute leukema aberration antigen expression． 5 out of 47 cases acutemyeliod leukemia patients accompany lymhocytic interrelated antigenexpression． 2/l5 cases acute lymphoid leukemia accompany myelocyteinterrelated antigen expression． 2 cases acute lmphoid leukemia are T cell and B cell interrelated antigen mingle expression． had been examined46,XY,t(8,2l) translocation of chromosome and bcr/abl fusion genes inthe acute leukemia patients． 16 out of 20 chronic myeloid leukemia patientshad philadelphia chromosome． 7 out of 20 patients had complicate karyotype． 5 out of 20 patients had bcr/abl fusion gene, l out of 4 patient had bcr/abl fusion gene that Ph chro mosome showed negative in CML． 3/4 cases patients had complicate chromosome． The ratio of CR use l time chemotherapy and the total ratio of CR using 2 times chemotherapywith aberration antigen expression in acute leukemia was significantly lessthan those of the acute leukemia patient had single system antigenexpression(P<0.05)． The time of CML-BC with complicate c hromosome karyotype was significantly short than those of Ph showed negative in CML(P<0.05)． Conclusion: The MICM classification is more acc urate for diagnosis of leukemia and more significant in guiding the leukemiatreamen．