Overall Estimation of Anti-Oxidant Activity by Mammal Macrophage

Super oxide anion had been known as a critical role in a phagocytic cell when microorganisms invade inside. However, this radical seldom need for a clean environment such in a developed country. Moreover, this agent sometimes triggered as serious factor in the site of inflammation. There are many papers which concerned newly found anti-oxidative substances. However, many reports only focused on the molecular basis in vitro, how oxidative stress damaged the human tissue. The key factor for oxidative stress needs to be discussed first for immunological standpoint, such as armed reduction treaty. In other words, minimum armed force is necessary for an invasion by outer or inner irregular cells. On the basis of phagocytic function of the cell in vivo, we tried to access anti-oxidative activity by employing peritoneal murine macrophage and human monocyte. So, the purpose of this study is to establish a conventional but tailor made system for which is useful for evaluating the anti-oxidative activity to different constition. Especially in human peripheral monocyte, the sample was tested each constitution such in granulocyte rich type and lymphocyte rich typ. In order to establish the direct estimation by phagocytic cell, a murine peritoneal exudates cell suspension was obtained after one week of oral administration with anti-oxidative sample. The samples we selected were traditional Chinese medicine, Shi-Quan-Da-Bu-Tang (SDT), Bu-Zong-Ye-Qi-Tang (BYT) and Shao-Chin-Rong-Tang sCRT), Agaricus, Chlorella, and Propolis. The amount of generated super oxide anion was calculated through measuring the optical density change of this supernatant using cytochrome-C which was reduced by generated super oxide anion in the phagocytic cell. The generated super oxide anion after administration of Agaricus, Chlorella, SDT, BYT, sCRT and Propolis were 2.64, 1.95, 1.24, 1.25, 2.88, 2.55 × 10﹣5 mmol/ml, respectively, whereas 2.85 × 10﹣5 mmol/ml in control group. The fermented sample from SDT, BYT, sCRT and microfied Propolis were also tested and the value of super oxide anion were 0.62, 0.84, 1.50 and 2.55 × 10﹣5 mmol/ml, respectively. In this report, we discussed the direct method for measuring the generated super oxide anion of macrophages, especially from different constitution.

DETERMINATION OF SERUM SOLUBLE MACROPHAGE COLONY-STIMULATING FACTOR RECEPTOR LEVELS IN PATIENTS WITH HEMATOLOGICAL DISEASES

Objective: To investigate the serum levels of soluble macrophage colony-stimulating factor receptor (M-CSFsR) in normal subjects and patients with hematological diseases and its clinical implications in hematological diseases. Methods: The concentration of M-CSFsR was determined by ELISA. The serum M-CSFsR was identified and characterized by immunoprecipitation and Western blotting. Results: The mean serum level of M-CSFsR of 123 normal individuals was 0.48 ng/ml ± 0.41 ng/ml. Immunoprecipitation and Western blotting assay revealed a ～ 90kD band of serum M-CSFsR. The mean serum M-CSFsR level of 60 patients with acute lymphoblastic leukemia (ALL), 36 patients with acute myeloblastic leukemia (AML), 13 patients with myelodysplastic syndrome (MDS) and 42 patients with aplastic anemia (AA) .were 0.22 ng/ml±0.23 ng/ml, 0.17 ng/ml±0.16 ng/ml, 0.19 ng/ml±0.16 ng/ml and 0.23 ng/ml±0.21 ng/ml, respectively, which were significantly lower than that of normal subjects (P=0.002 ,P<0.0001,P<0.0001 andP<0.0001). The mean serum M-CSFsR level of 51 idiopathic thrombocytopenic purpura (ITP) patients was significantly higher than that of normal subjects (2.05 ng/ml±2.75 ng/ml,P<0.0001). Conclusion: The serum M-CSFsR levels of patients with ALL, AML, MDS and AA were significantly lower, while the level of patients with ITP was significantly higher than that of normal individuals. Patients with severe ITP (platelet count<30×l09/L) had the highest M-CSFsR level. It suggested that the abnormal levels of serum M-CSFsR may associate with some hematological diseases and may contribute to the pathological process.

Rodent Macrophage Select Vin Blank Together with Vin Rouge According to SO Level <i>in Situ</i>

Phagocytic cells are known as multi potent activities for its ontogenical and phylogenetical aspect. One of the strike actions is super oxide anion known as critical role of the cell when microorganisms invade inside the cytoplasm. This agent sometimes triggered as a serious factor in the site of inflammation. There are many papers which concerned newly accessed anti-oxidative substances. However, many reports only focused on the molecular basis in vitro, suggesting vin rouge for their rich content of polyphonol rather than vin blank with a French paradox. We had been suggested that the key factor for oxidative stress needs to be discussed first for immunological standpoint, such as in phagocytic cell in situ. With this assay system in rodent macrophage, SO level was about the same for both groups administered vin blank as well as vin rouge.

Bone marrow-derived mesenchymal stem cells modulate autophagy in RAW264.7 macrophages via the phosphoinositide 3-kinase/protein kinase B/heme oxygenase-1 signaling pathway under oxygen-glucose deprivation/restoration conditions

Background: Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.Methods: We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditionsin vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.Results: The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20vs. 0.44 ± 0.08,t = 6.67,P < 0.05), while the expression of p62 was decreased (0.77 ± 0.04vs. 0.95 ± 0.10,t = 2.90,P < 0.05), and PI3K (0.40 ± 0.06vs. 0.63 ± 0.10,t = 3.42,P < 0.05) and p-Akt/Akt ratio was also decreased (0.39 ± 0.02vs. 0.58 ± 0.03,t = 9.13,P < 0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68 ± 0.14vs. 1.27 ± 0.20,t = 4.12,P < 0.05), up-regulated p62 expression (1.10 ± 0.20vs. 0.77 ± 0.04,t = 2.80,P < 0.05), and up-regulated PI3K (0.54 ± 0.05vs. 0.40 ± 0.06,t = 3.11,P < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05vs. 0.39 ± 0.02,t = 9.13,P < 0.05). A whole-genome microarray assay screened the differentially expressed geneHO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration ofHO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway.Conclusions: Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstancesin vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.

Aspect of QOL Assessment and Proposed New Scale for Evaluation

According to the reports from Ministry of Health in each country, the average life span is elongated over the past decade. This trend is expected to be keeping constant further in the future and at the same time continued efforts should be made. Next, it is necessary for us to consent about improving the quality of life. In an effort of improving the quality of life, other than the western medicine, we have attempted to introduce many traditional medical practices, including the oriental medicine, from various parts of the world into the medical field as a complementary & alternative medicine (CAM). Judging both positive and negative aspects by the evaluation standard of the Western medicine, we try to obtain numerical aspects in termed of QOL. Some methods of alternative medicine involved in this review are acupuncture, moxibustion, TCM and other traditional medicine. Above all, WHO suggests selecting the suzerain nation of acupuncture moxibustion, which has been developed in the oriental countries as a specialized cure, by identifying merits & demerits and ascertaining which nation can be a the best model for special field. At the moment for evaluating CAM, for example, what kind of methods is/are suitable for evaluating each CAM. We have been trying to propose the peripheral leukocyte, that is one of the best marker for evaluating CAM. Our trial is a hot spring hydrotherapy, acupuncture and moxibution, light exercise and Ondle/floor heating etc. In these trials, we find the common results from leukocyte effects which exhibit a strong correlation for the regulation in number and function, as a new scale for evaluating QOL. The contents are the result from the data showing the much in number of volunteer tend to down regulate, on the other hand, lower number of volunteer is up-regulated at next day by each menu of CAM as a constitution dependent manner. However, even in western medical methodology, double blind and cross over system are the better way to evaluation even in CAM. The responsibility in human to each CAM therapy is different in individually, so to say according to constitution. In other words, there are so many vectors within the group, including at least three groups, up-regulating, down regulating and stand stilled one. So we have no positive results by the method by sum-up and made mean. According to this system, individual vector is cancelled each other and real effect is not exhibited as a final result. So, we would like to present more smart way to access the efficacy for CAM menu. That is trial to plot each individual effect on the linear function and making slant/co-efficiency that plot each variable vector which is derived from the relative value compared to that of the day before. This procedure is easy to compare between each impact to the volunteer. We would like to propose this evaluation system as Super Constitution System: SCS. This kind of regulation showed within a 24 hours, for the leukocyte subsets, granulocyte and lymphocyte are changed even if they are under control of the circadian rhythm. So, for the purpose of evaluating leukocyte deviation, we set the point for evaluation at the same time-zone from the first set of evaluation to the next time-zone of evaluation. Under these conditions, we get a same result reproductively that the whole number of leukocyte, and its subset, granulocyte and lymphocyte also regulate within a 24 hours. This kind of phenomenon is the case that we confirm a lot of kind of CAM with repetition. This kind of regulation is confirmed in the content of the emotional hormone regulation. Within a hormonal change, adrenalin and dopamine are just under the regulation in the mode of SCS. However, the life span of leukocyte are at lease 3 - 4 days in rapid group of leukocytes, and no such a drastic apoptosis is induced by such a stimulation. For the purpose of scientific explanation, we also propose that the emotional hormone is concerned this change of leukocytes population. For the results of this hypothesis, there are reasonable changes in the peripheral blood about emotional hormone, adrenaline and dopamine. Other hormone concern thyroid grand is not changed significantly. In the final part of this chapter, we also propose that the suitable approach for anti-oxidative assessment and the effective pathway of complement for enabling some historical food supplement activate the whole body cell that expresses a complement receptor. This is not a case of infection but another stage of complement activation by fragmented polysaccharides for historical human use as foods. The protein materials are also fermented and served as digestive food around the world. Almost all the products have some distinctive smell but accompanying some effects through independent digestive rout for intact protein itself. With these aspects, we try to emphasize SCS and discuss by showing evident based manner with various menu of CAM.

Reconstruction of hematopoietic function in irradiated mice promoted by polysaccharide peptide

Objective: To study the effect of polysaccharide peptide (PSP) on the reconstruction of the hematopoietic function in mice irradiated by lethal dose. Methods: The characteristics of proliferation of colony forming unit granulocyte macrophage (CFU GM) and colony forming unit spleen (CFU S) were measured after continuous injection of PSP in mice for seven days with different doses. Results: Injection of 25 mg/kg PSP in mice could promote and increase CFU GM proliferation and cell mitosis, markedly enhance survival ratio, survival time and CFU S. Conclusions: PSP has significant regulative effects on the reconstruction of the hematopoietic and the immunologic functions in mice irradiated by lethal dose.