| 1 Objective:In this study,we first evaluated and analysed the curative effect of Shugan Jianpi Huoxue Method Combined with Western Medicine for the treatment of liver cancer.Main outcome measures: including the evaluation of curative effect in the treatment efficiency,the change of KPS score rate,the function of immune cells.On the basis of evidence-based medicine,we investigated the influence of medicated plasma containing Beilong Ruangan decoction on proliferation of peripheral blood dendritic cells from HBV associated hepatocellular carcinoma patients,amied at providing a new way of thinking: “fom evidence-based medicine,basic laboratory research then to clinical application" applied Chinese medicine for the treatment of hepatocellular carcinoma,and providing the exact basis for Chinese in clinical application2 Methods2.1 Meta analysis of curative effect of Traditional Chinese Medicine combined with Western Medicine on liver cancerWe used Cochrane system evaluation method,computer retrieval CNKI Chinese CNKI,cqvip Chinese journal database.The retrieval period is from 1990 to 2016 years.Clinical studies on Shugan Jianpi Huoxue Decoction Combined Western Medicine treatment of primary liver cancer.Using statistical software Rev Man5.3,the curative effect after the treatment,the patient's quality of life score(KPS),and cellular immune function(T cell subsets and NK cells)were statistically analyzed with Meta.2.2 The influence of medicated plasma containing Beilong Ruangan decoction on proliferation of peripheral blood dendritic cells from HBV associated hepatocellular carcinoma patients2.2.1 The preparation of medicated plasma containing Beilong Ruangan decoctionSPF rats were randomly divided into 3 groups,blank plasma group,Beilong Ruangan Decoction low dose group,and Beilong Ruangan medium dose group,rat gastric lavage volume according to the surface area for conversion algorithm,low dose equivalent to 6.562 times of adult clinical dosage,medium dose equivalent to 5 times of low dose,the blank plasma group with equal amount of saline,gavage twice a day for 5 days,at 2h after the last gavage,intraperitoneal injection of 10%chloral hydrate anesthesia(0.35ml/100g),to be completely open after anesthesia,abdominal aortic blood separation,holding the needle into the abdominal aorta blood connection,3000r/min centrifugal 15 min,under normal temperature,0.22 um membrane filtration,the plasma will be divided into 1.5ml EP sterile tube,and-20? the refrigerator spare.2.2.2 Culture and identification of DCs in peripheral bloodSterilely from arm vein peripheral blood 20 ml,heparin sodium anticoagulation(50U/ml),PBS 1:1 diluted,lymphocyte separating medium peripheral mononuclear cells of PBMCs,PBS DMEM,washed 2times with which washing 1 times.The concentration of PBMCs FCS DMEM's 10% solution adjustment 2 x 106 /ml into 6 well culture plates,each hole 2.5ml,37 ?,5%CO2 incubator overnight incubation,the supernatant liquid suction abandon DMEM,37?preheating gently washing plates 2 times,the removal of non adherent cells,obtain adherent mononuclear cells.Granulocyte macrophage cells were added stimulating factor(GM-CSF)shot down 1000U/ml IL-4 500U/ml complete medium,the next day was half changed and cytokines,cultured for fifth days and seventh days with tumor necrosis factor alpha(TNF-alpha)20ng/ml,collected ninth days suspended cells to mature DCs were observed by morphological identification.2.2.3 The detection proliferation of DCs HBV associated hepatocellular carcinoma patients 10 cases as experimental group,each arm venous blood 20 ml,DCs were isolated and cultured,collected and cultured for fifth days with DCs,medicated plasma containing Beilong Ruangan decoction intervention for 12 h,24h and 48 h,by CCK-8 reagent,detection of cell proliferation.3 Result3.1 The result of Meta analysis :Traditional Chinese Medicine treatment of liver and spleen and activating blood circulation combined with Western Medicine treatment of liver cancer compared with pure Western Medicine treatment,the treatment group can obviously improve the short-term effect,quality life of patients and the function of immune cells,and it showed a stastic difference(P<0.01)3.2 Plasma containing Beilong Ruangan decoction can stimulate the proliferation of peripheral blood dendritic cells from HBV associated hepatocellular carcinoma patients,which depended on time and concentrations,24 h low dose concentrations of 5%?10%?20%,48 h low dose concentrations of 5%? 10%? 20% and 48 h low dose concentration of 20% showed the significantly difference compared the blank plasma groups(P<0.01),and the 48 h low dose concentrations of 20% proved the best effect on the proliferation of dendritic cells.4 Conclusions4.1 Through the Meta analysis of Traditional Chinese Medicine treatment of liver and spleen and activating blood circulation combined with Western Medicine treatment of liver cancer compared with pure Western Medicine treatment,can significantly improve the short-term effect after treatment,improve the quality life of patients,enhance the immune function of cells.4.2 Compared with healthy people,the number of DCs in peripheral blood of patients with HBV related liver cancer was significantly reduced and immune dysfunction.4.3 The plasma containing Beilong Ruangan decoction can stimulate proliferation of peripheral blood dendritic cells from HBV associated hepatocellular carcinoma patients,dignosed of liver depression with spleen deficiency syndrome,and the low dose concentration of 20%interved 48 h was the best.4.4 Beilong Ruangan decoction belongs to the Shugan Jianpi method,preliminary clinical follow certificate of medical and basic experiment,the party in improving the quality of life of patients with liver cancer,curative effect of immune function,and is worthy of promotion and use. |
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