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Objective To investigate the inhibitory effect and mechanism of Metformin (Met) combined with irradiation in CT26WT cell lines or mouse models with transplanted tumors.Methods CT26WT cell line was treated with 0.5 μmol/L,1.0 μmol/L,5.0 μmol/L and 10.0 μmol/L Met,and CellTiter Glo kit was used to detect the inhibitory effect of Met at different concentrations on the viability of CT26WT cells.CT26WT cell line was treated with the control,Met (10 pmol/L),15 Gy irradiation and 15 Gy irradiation+Met (10 μmol/L).Clone formation assay was employed to detect the cell proliferation activity.Bablc mouse models of transplanted tumors (tumor size＞ 150 mm3) were established and randomly divided into the control,15 Gy irradiation,Met and 15Gy irradiation+Met groups.Mice were given with 750 mg/kg Met at 24 h before irradiation.Transplanted tumor volume was measured regularly to delineate the growth curve of transplanted tumors and survival curve.The expression levels of P-H2AX and Sting proteins in CT26WT cells and transplanted tumors were detected by Western blot.The infiltration of CD8a (+) T cells in transplanted tumor tissues was detected by immunohistochemistry.Results The relative cell survival rate was 100％,87.9％,87.8％,87.3％ and 76.5％ in the 0,0.5,1.0,5.0 and 10.0μmol/L Met groups,respectively (all P＜0.05).The inhibitory effect of 10.0 μmol/L was significantly stronger than that of 5.0 μmol/L (P＜0.001).The colone formation rate 34.0％,24.0％,22.3％ and 14.0％ in the control,Met,15 Gy irradiation,Met+ 15Gy irradiation groups,respectively (all P＜0.001).Western blot showed that compared with the control group,the expression of Sting protein was increased by 2.99-fold after Met treatment (P＜0.001),and increased by 1.37-fold and 4.41-fold in the 15 Gy irradiation and 15Gy irradiation+Met groups (both P＜0.01).Compared with the 15 Gy irradiation group,the expression of P-H2AX protein was significantly increased by 1.43 times after treatment with 15Gy+Met (P＜0.001).The transplanted tumor growth curve showed that the transplanted tumor growth in the 15 Gy+Met group was slower than that in the control group[(1007.0± 388.5) mm3 vs.(2639.0± 242.9) mm3,P＜ 0.05)].The overall survival time in the 15 Gy irradiation+Met group was 48 d,significantly longer than 32 d in the control group (P＜0.001).Compared with the control group,the expression of P-H2AX and Sting proteins in the 15 Gy+ Met group was increased by 8.8-fold and 1.6-fold (both P＜0.001).Immunohistochemical staining showed that the infiltration of CD8a (+) T cells in the 15 Gy irradiation+Met group was significantly higher than that in the control group (P＜0.01).Conclusions Met combined with radiotherapy can inhibit the proliferation and clone formation of colon cancer cells,probably by aggravating DNA damage and activating the Sting signaling pathway,eventually leading to the increase of CD8a (+) T cells in tumor tissues and enhancing the killing effect upon transplanted tumor cells.
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Objective:To establish an HPLC method for the simultaneous determination of minoxidil and tretinoin in compound minoxidil gel. Methods:A Shim-Pack VP-ODS C18(250 mm × 4. 6 mm,5 μm)column was used with the mobile phase of methanol and 2% acetic acid solution by gradient eleution . The detection wavelength was respectively set at 280 nm and 350 nm,and the flow rate was 1. 0 ml·min -1 . The column temperature was 35℃ and the injection volume was 10 μl. Results:Minoxidil and tretinoin had a good linear relationship within the range of 4. 0- 240. 0 μg·ml-1 and 0. 05- 3. 00 μg·ml-1(r =0. 999 6 and 0. 999 3),respectively. The average recovery was 99. 56%(RSD = 0. 42% ,n = 9)and 99. 36%(RSD = 0. 50% , n = 9),respectively. Conclusion:The established method is simple,and suitable for the simultaneous determination of the two constituents in compound minoxidil gel.
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ObjectiveTo explore the effect of celecoxib on the tendon adhesion and healing after anastomosis.Methods54 New Zealand white rabbits were randomly assigned to one of 3 ( celecoxib,ibuprofen,and saline) groups.The deep flexor tendon was transected,followed by a primary repair.The care was begun the day after surgery and was continually provided for 14 days.Celecoxib was given[20 mg/( kg · d) ],ibuprofen was given [75 mg/( kg · d) ],or the same volume of saline solution was given respectively.At the 4th and 8th week the animals were killed and assessed by general observation,histologically observation,and biomechanical testing.ResultsGeneral observation,celecoxib group and ibuprofen group showed good shape of tendons,smooth surface,and covered with a film - like tissue,and it was easy to be separated.Saline group showed that tendon and surrounding tissue medium had dense adhesions,and it was difficult to be separated.Histologically observation showed between tendon and paratenon had clear gap in celecoxib and ibuprofen group.The mature fiber cells gradually arranged rules and in the same direction.Saline between the tendon and paratenon gradually emerged a gap,but more fibroblasts and arranged in irregular.Biomechanical testing showed that tendon sliding resistance appeared in celecoxib and ibuprofen group.Compared with saline group,the differences were statistically significant (0.354 ± 0.078/0.382 ±0.121 vs 0.521 ±0.126,P ＜0.05;0.075 ±0.035/0.097 ±0.043 vs 0.414 ±0.110,P ＜0.01).UTS (ultimate tensile strength had statistical significance among celecoxib,saline groups and ibuprofen group.(36.812 ±6.388 vs 24.899 ±4.667,P ＜0.05;34.297 ±8.132 vs 24.899 ±4.667,P ＜0.01 ;54.515 ±4.688/59.037 ± 6.606 vs 42.418 ± 5.594,P ＜ 0.01 ).ConclusionsCelecoxib can effectively prevent tendon adhesion,and does not affect the tendon healing.
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Objective To detective the carcinogenesis and operation principle of cyst canceration after internal drainage(ID) operation for congenital choledocal cyst (CCC).Methods The clinical data of 25 patients with cyst carcinoma after ID operation for CCC in the past 28 years were analysed retrospectively.Results The total canceration rate after internal drainage of CCC were 30.49%(25/82); after cystoduodenostomy was 35.29%(14/51),after cystojejunostomy was 22.58%(11/31), respectively. In the 25 cases, three of them were operated with Wipple operation , 4 with tumour resection plus biliary reconstrustion operation, 4 local resection with external drainage ,14 with external drainage only. Conclusions Internal drainage of CCC should be aborted becaus of the high canceration rate after the operation.