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Objective To investigate the effect of fractionated radiotherapy on the immune system of mice with subcutaneously transplanted hepatocellular carcinoma.Methods Logarithmic growth of mouse hepatocellular carcinoma cells Hepa 1-6 were inoculated subcutaneously on the right side of C57BL/6 J mice (1×107 cells/mice).The tumor-bearing mice were randomly divided into control group (Ctrl) and irradiation group (IR),20 mice in each group.Additionally,10 healthy mice were set as normal control group.Local fractionated X-ray irradiation of 8 Gy×3 fraction was given to the subcutaneous tumors,and the dose rate was 0.883 Gy/min.At 7 and 14 d after irradiation,the tumor organ index,spleen organ index,spleen pathological changes,and splenic T lymphocyte subsets,B lymphocyte subsets,and NK cells were detected.Results Compared with Ctrl,at 7 and 14 d after irradiation,the tumor organ index decreased (t =4.649,26.34,P＜0.05),and the percentage of NK cells increased significantly (t =3.952,3.633,P＜0.05).The percentages of CD3+,CD4+,CD3+CD4+ lymphocytes and the ratio of CD4+/CD8+ lymphocyte decreased at 7 d after irradiation (t=3.193,3.656,3.219,2.641,P＜0.05),and the percentage of CD3+lymphocyte decreased at 14 d after irradiation (t =3.031,P＜0.05).But after irradiation,there were no significant changes in spleen organ index,B lymphocyte,CD3 + CD4+ lymphocyte and CD8 + lymphocyte.Conclusions Local hepatoma radiotherapy causes imbalance of lymphocytes in distal spleen of mice and hence reduces immunity,which provides a novel mechanism of radiological immunity damage.
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Objective@#To investigate the effect of fractionated radiotherapy on the immune system of mice with subcutaneously transplanted hepatocellular carcinoma.@*Methods@#Logarithmic growth of mouse hepatocellular carcinoma cells Hepa 1-6 were inoculated subcutaneously on the right side of C57BL/6 J mice (1×107 cells /mice). The tumor-bearing mice were randomly divided into control group (Ctrl) and irradiation group (IR), 20 mice in each group. Additionally, 10 healthy mice were set as normal control group. Local fractionated X-ray irradiation of 8 Gy×3 fraction was given to the subcutaneous tumors, and the dose rate was 0.883 Gy/min. At 7 and 14 d after irradiation, the tumor organ index, spleen organ index, spleen pathological changes, and splenic T lymphocyte subsets, B lymphocyte subsets, and NK cells were detected.@*Results@#Compared with Ctrl, at 7 and 14 d after irradiation, the tumor organ index decreased (t=4.649, 26.34, P<0.05), and the percentage of NK cells increased significantly (t=3.952, 3.633, P<0.05). The percentages of CD3+ , CD4+ , CD3+ CD4+ lymphocytes and the ratio of CD4+ /CD8+ lymphocyte decreased at 7 d after irradiation (t=3.193, 3.656, 3.219, 2.641, P<0.05), and the percentage of CD3+ lymphocyte decreased at 14 d after irradiation (t=3.031, P<0.05). But after irradiation, there were no significant changes in spleen organ index, B lymphocyte, CD3+ CD4+ lymphocyte and CD8+ lymphocyte.@*Conclusions@#Local hepatoma radiotherapy causes imbalance of lymphocytes in distal spleen of mice and hence reduces immunity, which provides a novel mechanism of radiological immunity damage.
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OBJECTIVE：To study the effects of gin seng root extract on autophagy ，proliferation and cytokine of splenic lymphocyte of mice ，and to study its mechanism. METHODS ：The splenic lymphocyte of mice were divided into blank control group，positive control group （concanavalin A ，5 μg/mL），different concentration of ginseng root extract groups （32，125，500 μg/mL，by lyophilized powder ）. After 48 h of culture with the corresponding medicine ，acridine orange staining method was used to detect autophagy of splenic lymphocyte ；CCK-8 assay was used to detect the cell proliferation ；ELISA assay was used to determine the levels of IL- 4，IL-6 and TNF-α in cell culture；Western blotting method was used to detect the expression of LC 3B and Beclin- 1，as well as the phosphorylation of AMPK ，AKT and mTOR. RESULTS ：Compared with blank control group ，32， 125，500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice（P＜0.05 or P＜0.01）；125，500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes，the levels of IL- 4，IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ，significantly increased the protein expression of Beclin- 1， quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ，2015，23（3）：232-243. the phosphorylation of AMPK protein ，and significantly reduced the phosphorylation level of AKT and mTOR proteins ，with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS ：Ginseng root extract can induce autophagy of mice splenic lymphocytes，activate the activity of splenic lymphocytes ，regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ，so as to exert immune enhancement effect.
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Objective To explore the immune mechanism in the systemic lupus erythematosus MRL/lpr mice at different months of age, and to provide the basis for research of its pathogenesis. Methods 3-,4-,5- and 6-month old female MRL/lpr mice, and wild type C57 female mice were used in this study, 10 mice per each group. Their organ coefficients were determined. ELISA was performed to detect the serum levels of double stranded DNA(ds-DNA) antibody. The interleukins IL-2,IL-4,IL-17 and tumor necrosis factor-α(TNF-α)in spleen tissue were detected. Flow cytometry was used to assess the content of spleen lymphocyte CD3 cells and CD4/CD8 cell ratio. Results Compared with the 3-month old wild-type C57 mice,the spleen coefficient,the blood concentration of ds-DNA antibody,IL-2 and TNF-α in the 3- to 6-month old MRL/lpr mice were significantly increased(P < 0.05). There was no significant difference between the concentrations of interleukin IL-4(P> 0.05). The blood concentration of IL-17 in the 5- and 6-month old MRL/lpr mice was significantly lower(P < 0.05 or P < 0.01)than that in the 3-month old MRL/lpr mice. The rest indexes of MRL/lpr mice showed no obvious changes or significant difference in the mice at different ages. Compared with the 3-month old C57 mice,the spleen CD3 lymphocyte concentration in the MRL/lpr mice was significantly decreased(P< 0.01). With the increasing age, the CD3 lymphocyte concentration and D4 +/CD8 +cell ratio in the MRL/lpr mice were decreased, however, showing a non-significant difference(P ﹥0.05). Conclusions The data obtained in this study indicate that 3-month old lupus MRL/lpr mice have already immune injury,increasing with the increase of age of the mice.
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Objective:To probe e ect of a Chinese compound recipe of nourishing yin and clearing heat and prednisone on nuclear protein of splenic lymphocytes in mice.Methods:MRL/LPR female lupus mice were divided into four groups respectively treated with isotonic Na chloride,traditional Chinese medicine,Western medicine and integrated Chinese and Western medicine,then nucleoproteins of splenic lymphocytes were extracted and detected by SELDI-TOF-MS method.Result:There were some proteins in control group which aggravated autoimmune course,and some proteins in Western medicine group which rivalried immunological activity;integrated Chinese and Western medicine and Western medicine showed similarly e ect on nucleoprotein of splenic lymphocytes.Conclusion:The recipe of nourishing yin and clearing heat and prednisone can treat the disease by uencing in the di erentiation of splenic lymphocytes and the expression of proteins.
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Objective To explore the molecule mechanisms of migraine with hyperactivity of liver-yang related proteins. Methods Animal model of migraine with hyperactivity of liver-yang was established through electrical trigeminal ganglion stimulation,and by administering orally with Fuzi decoction. The total proteins of splenic lymphocytes in the rats were separated by immobilized pH gradient-based two-dimensional gel electrophoresis. With coomassie blue staining,the 2-DE images were analyzed by PDQuest 7.0 software. Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) and database were used to identify the differential proteins. Result Ten differential spots protein were identified,and they were chloride intracellular channel 1,thioredoxin peroxidase Ⅰ,thioredoxin peroxidase Ⅱ,sequence 7 from patent WO9709348,Rho GDP dissociation inhibitor (GDI) alpha,actin related protein 2/3 complex neurofilament light polypeptide,proteasome subunit beta type and heat shock protein-27,annexin A1. Conclusion These protein may be involved in the development of migraine with hyperactivity of liver-yang. These data provide useful clues for elucidating the mechanisms of migraine with hyperactivity of liver-yang.
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The water-soluble polysaccharide PNM isolated from the submerged culture of Phellinus nigiricans by hot water extraction and ethanol precipitation, was purified by freeze thawing, methods of Sevag, and then fractionated by Sepharose CL-6B gel filtration chromatography, giving a polysaccharide fraction named as PNMⅠ. HPLC analysis showed that PNMⅠwas a homogeneous polysaccharide, with a weight-average molecular weight of 29 kDa. GC analysis indicated that PNMⅠis a heteropolysaccharide composed of mannose, galactose and glucose with a ratio of 0.35:0.44:1.00. Pharmaceutical experiments showed both PNM and PNMⅠ have a significant effect on the immunomodulatory activities of murine splenic lymphocytes in vitro. Also PNMⅠ could inhibit the growth of sarcoma 180 solid tumor in vivo.
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The method for measuring glucocorticoid hormone receptors in rat splenic lymphocytes was developed. The results showed that there were specific binding sites of 3H-Dexamethasone with high affinity (Kd : 5,05?1.70 nmol/L), low binding capacity (Bm3s or R0 : 6773?585 specific binding sites/cell) and tempera- ture-dependent nuclear translocation in the rat splenic lymphocytes. This paper provides a convenient method for study of the molecular mechanisms of glucocor-ticoid hormone action and pathophysiology of glucocorticoid hormone receptors.