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OBJECTIVE:To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC- 3 cells. METHODS:Using human pancreatic cancer BxPC- 3 cells with natural deletion of Smad4 gene as reaserch objects ,CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of BxPC- 3 cells after treated with different doses of metformin(5,10,20 mmol/L)for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test the migration of cells after treated with different doses of metformin (10,20 mmol/L)for 24 h. The number of migrating cells was recorded. qRT-PCR and Western blotting assay were performed to determine mRNA and protein expression of E-cadherin ,Vimentin and RGC- 32 in cells. RESULTS :Compared with control group and 5 mmol/L metformin group ,survival rate of cells were decreased significantly in 10,20 mmol/L metformin groups ,while apoptosis rate was increased significantly ;the apoptosis rate in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group (P<0.05). Compared with control group , the number of migrating cells was decreased significantly in 10,20 mmol/L metformin groups ,and the 20 mmol/L metformin group was significantly lower than 10 mmol/L metformin group (P<0.05). Relative mRNA and protein expression of E-cadherin were increased significantly in 10,20 mmol/L metformin groups ,and relative mRNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L metformin group ,relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group ,relative mRNA and protein expression of RGC- 32 in 10,20 mmol/L metformin groups were decreased significantly ;relative mRNA and protein expression of Vimentin as well as mRNA expression of RGC- 32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L metformin group (P<0.05 or P<0.01). CONCLUSIONS :Metformin can inhibit the proliferation and migration of pancreatic cancer cells through smael-independent pathways in a dose- dependent manner ,and promote their apoptosis ,which is associated with the inhibition epithelial- mesenchymal transition and the expression of RGC- 32 of pancreatic cancer.
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Objective To establish a method for determination of the content of Forsythoside A in Yinqiaosan granules . Methods HPLC method was adopted to carry on the determination .Welch Ultimate XB‐C18 column (250 mm × 4 .6 mm ,5μm) was used with the mobile phase of methanol‐0 .2% Glacial acetic acid solution by gradient elution . The flow rate was 1 .0 ml/min ,the detection wavelength was 330 nm ,and the column temperature was 20 ℃ .Results Forsythoside A showed a good linear relationship at the range from 0 .080 3~3 .211 μg (r=1 .000) .The average recovery rates were 103 .78% ,and RSD=0 .82% .Conclusion The method was convenient and accurate .Meanwhile ,it also has good specificity and reproducibili‐ty .It can be used effectively for the quality control of Yinqiaosan granules .
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Objective To introduce the application and assess the outcome of ProPlan CMF software in virtual surgery of mandibular resection and reconstruction with vascularized fibular graft.Methods Nineteen patients were performed mandibular resection,and immediate reconstruction with free fibular graft by application of ProPlan CMF software between March 1,2013 and January 31,2014,including 7 cases of osteoradionecrosis,6 cases of adamantoblastoma,4 cases of carcinoma of gingiva,2 cases of intra-osseous carcinoma of mandible.There were 12 males and 7 females,with an age range of 21-73 years (median,54 years).In each case,three-dimensional spiral CT scan of mandible and fibula was obtained before operation.The CT data were imported into the ProPlan CMF software and the virtual surgical planning was performed.After that,the mandibular rapid prototyping and surgical guide plates were made according to customized design.The segmental resection of mandible,titanium plate bending,fibular molding,insetting and fixing were then carried out by using these surgical guide plates.Results Operations were undertaken successfully according to preoperative design in all nineteen patients and negative margin was detected by intraoperative frozen section.The fibula was cut,shaped and fixed accurately.The operations took 4.5-7.0 hours (mean,5.5 hours).Primary healing of incision was obtained without complications except fistula in 2 cases of osteoradionecrosis,which were resolved with local wound care after 4 weeks.19 patients were followed up between 3 months to 1 year and all patients were alive with disease free.All patients were satisfactory with their maxillofacial appearance,occlusion and function.The ranges of mouth opening was 2.0-4.0 cm.The postoperative panoramic radiographs or CT scan showed perfect situation and contour of fibular graft.Healing of the donor site occurred without significant complications.All patients started early postoperative ambulation,and walked normally after 2 or 3 months.Conclusion The application of ProPlan CMF software in virtual surgical design and guide plates creation can simplify the operation,and improve the accuracy of mandibular resection and reconstruction,especially the spatial relationship of the mandible and the fibula graft.It is worth promotion widely in clinical practice.