高效液相色谱法测定免疫功能型保健食品中8种功效成分

目的建立高效液相色谱法同时测定免疫功能型保健食品中8种功效成分(左旋肉碱、L-茶氨酸、天麻素、吡啶甲酸铬、咖啡因、淫羊藿苷、菊苣酸、10-羟基-α-癸烯酸)的分析方法。方法样品用70%甲醇超声提取,采用C18色谱柱分离,以甲醇-磷酸二氢钾(0.1%磷酸)水溶液为流动相,梯度洗脱,外标法定量。结果左旋肉碱、L-茶氨酸在10.0~150.0 mg/L,吡啶甲酸铬、天麻素、咖啡因和淫羊藿苷在1.5~30.0 mg/L,菊苣酸在5.0~100.0mg/L,10-羟基-α-癸烯酸在2.5~50.0mg/L质量浓度与色谱峰面积有良好的线性关系,相关系数r≥0.9990。方法检出限为1~10mg/kg,回收率为93.8%~108%,相对标准偏差(relativestandarddeviations,RSDs)为0.7%~6.8%。结论该方法快速、准确、灵敏,适合保健食品中8种增强免疫力功效成分的同时测定。

Influence of epigallocatechin gallate on the immune function of dendritic cells after ultraviolet B irradiation

To investigate the protective effect of epigallocatechin gallate （EGCG） on the immune function of dendritic cells （DCs） after ultraviolet B irradiation （UVB） and its underlying mechanisms, the monocytes were isolated from peripheral blood and cultivated into DCs with cytokines, such as GM-CSF and IL-4. DCs were harvested after cultivation for 7 d and subjected to irradiation with different dosages of UVB. Then, 200 μg/ml of EGCG were added in certain groups immediately after irradiation. DCs simply treated with UVB or treated with both UVB and EGCG were co-cultured with lymphocytes, and MTT assay was used to detect the ability of DCs to stimulate proliferation of lymphocytes. Surface markers CDS0, CD86, HLA-DR and CD40 were detected by flow cytometry, and the levels of IL-10 and IL-12 secreted from DCs 2d h after cultivation were measured by ELISA. It was demonstrated that UVB irradiation could inhibit the ability of DCs to stimulate the proliferation of lymphocytes and surface expressions of CDS0, CD86, HLA-DR and CD40 on DCs in a dose-dependent manner. The inhibition rate of DCs was improved to some extent after treatment with 200μg/ml of EGCG. When the concentra- tion of EGCG exceeded 100 μg/ml, the enhancing effect of EGCG on the expression of the co-stimulating molecules on DCs could be demonstrated in a dose-dependent manner. UVB showed no significant influence on the secretion of IL-10 and IL-12 from DCs, while EGCG could down-regulate the secretion level of IL-12 and up-regulate that of IL-10. It is concluded that EGCG can antagonize the inhibitory effect on DCs induced by UVB irradiation. This function has some relationship with its protecting effect of the expression of the co-stimulating molecule on the surface of DCs and the secretion level of IL-10 and IL-12.

Experimental-Biological Model of Bartonellosis

Bartonella are known to be important causes ofzooanthroponotic diseases. The range of human infection varies from mild lymphadenopathy and asymptomatic bacteremia to life-threatening systemic disease in immunocompromised patients. Microbiological improvements in isolation methods and PCR amplification of organism-specific DNA sequences have resulted in a dramatic increase in reports describing human patients with bartonellosis. Nevertheless, clearly and successful isolation ofBartonella spp. from bacteremic animals and human patients remains an ongoing challenge. Technology of experimental bartonellosis due to intraperitoneal introduction of biological material samples containing causative agents to laboratory animals is presented in the article. White nonlinear mice with the artificially cyclophosphamide formed immunodeficient state may be used as an experimental model for further investigation of the biological alterations responsible for angiomatosis. On the other hand, we believe that this new method will enhance the diagnostic sensitivity and specificity needed to achieve a diagnosis of bartonellosis.

Inflammatory models drastically alter tumor growth and the immune microenvironment in hepatocellular carcinoma

Initiation and progression of hepatocellular carcinoma （HCC） is intimately associated with a chronically diseased liver tissue. This diseased liver tissue background is a drastically different microenvironment from the healthy liver, especially with regard to immune cell prevalence and presence of mediators of immune function. To better understand the consequences of liver disease on tumor growth and the interplay with its mi- croenvironment, we utilized two standard methods of fi- brosis induction and orthotopic implantation of tumors into the inflamed and fibrotic liver to mimic the liver condition in human HCC patients. Compared to non-diseased con- trols, tumor growth was significantly enhanced under fi- brotic conditions. The immune cells that infiltrated the tumors were also drastically different, with decreased numbers of natural killer cells but greatly increased num- bers of immune-suppressive CDllb^＋ Gr1^hi myeloid cells in both models of fibrosis. In addition, there were model- specific differences： Increased numbers of CD11b^＋ mye- loid cells and CD4^＋ CD25^＋ T cells were found in tumors in the bile duct ligation model but not in the carbon te- trachloride model. Induction of fibrosis altered the cytokine production of implanted tumor cells, which could have far- reaching consequences on the immune infiltrate and its functionality. Taken together, this work demonstrates that the combination of fibrosis induction with orthotopic tumor implantation results in a markedly different tumor mi- croenvironment and tumor growth kinetics, emphasizing the necessity for more accurate modeling of HCC pro- gression in mice, which takes into account the drastic changes in the tissue caused by chronic liver disease.

Effect of cold-dryness on pulmonary and immunologic function in chronic obstructive pulmonary disease model rats

OBJECTIVE： To study the effects of cold-dryness on pulmonary and immunologic function of peripheral T-lymphocytes in chronic obstructive pulmonary disease （COPD） model rats, and to provide references for the prevention and treatment of cold-dryness COPD in the Xinjiang region. METHODS： The COPD model was established with an elastase drip into the trachea combined with smoking. The cold-dryness COPD model was developed by stressing with a cold-dry environment. Success of the model was determined by observation of pathologic lung sections. Rats were sacrificed by exsanguination from the femoral artery and changes of peripheral blood CD4＋, CD8＋, and CD4＋/CD8＋ were detected by flow cytometryo Data were analyzed with SAS 11.5 statistical software. RESULTS： On the ninetieth day after ending the ex- periment, Peak expiratory flow in the cold-dryness COPD group was lower than that in the COPD and normal control groups （P〈0.01）. The time of inspiration in the cold-dryness COPD group was higher than that in the COPD and normal groups （P〈0.05）. Time of expiration （Te） in the cold-dryness COPD group was higher than that in the COPD and normal groups （P〈0.01）. 50% tidal volume expiratory flow （EFS0） in the cold-dryness COPD group was lower than that in the COPD and normal groups （P〈 0.01）, and EFS0 in the COPD group was lower than that in the normal group （P〈0.05）. CD4＋ content of peripheral blood in the cold-dryness COPD group was lower than that in the COPD and the normal groups （P〈0.05）. CD8＋ content in the cold-dryness COPD and COPD groups was higher than that in the normal control group （P〈0.01）, and CD8＋ content in the cold-dryness COPD group was higher than that in the COPD group （P〈0.01）. CD4＋/CD8＋ in the cold-dryness COPD group and the COPD group was lower than that in the normal control group （P〈0.01）, and CD4＋/CD8＋ in the cold-dryness COPD group was lower than that in the COPD group （P〈0.05）. CONCLUSION： In the cold-dryness COPD model, CD8＋ increased and CD4＋/CD8＋ decreased. Moreover, cold-dryness may aggravate this state. The effects of cold-dryness on pulmonary function main- ly manifested as prolongation of Te and decrease of EF50, which could be one of causes of cold-dryness environment in the northwest of China leading to COPD with region characteristics.