OBJECTIVETo investigate the effects and mechanisms of cordycepin,an effective component of cordyceps militaris, on renal interstitial fibrosis (RIF) and its related eIF2α/TGF-β/Smad signaling pathway.

METHODFirstly, 15 C57BL/6 mice were randomly divided into 3 groups,the control group (Group A), the model group (Group B) and the cordycepin-treated group (Group C). After renal interstitial fibrotic model was successfully established by unilateral ureteral obstruction (UUO), the mice in Group C were intraperitoneally administrated with cordycepin(5 mg x kg(-1) d(-1)) and the ones in Group A and B were administrated with physiological saline for 5 days. At the end of the study, the obstructed kidneys were collected and detected for the pathological changes of RIF, and the mRNA expressions of collagen type I (Col I) and α-smooth muscle actin (α-SMA) in the kidney by Northern blot. Secondly, after renal tubular epithelial (NRK-52E) cells cultured in vitro were exposed to transforming growth factor (TGF) -β with or without cordycepin, the mRNA expressions of Col I and collagen type IV( Col IV) by Northern blot, and the protein expressions of eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α ( p-eIF2α), Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) were tested by Western blot.

RESULTIn vivo, cordycepin alleviated RIF in model mice, including improving fibrotic pathological characteristics and mRNA expressions of Col I and α-SMA. In vitro, cordycepin induced the high expression of p-elF2α, and inhibited the expressions of p-Smad2/3, Col I and Col IV induced by TGF-β in NRK-52E cells.

CONCLUSIONCordycepin attenuates RIF in vivo and in vitro, probably by inducing the phosphorylation of eIF2α, suppressing the expression of p-Smad2/3, a key signaling molecule in TGF-β/Smad signaling pathway, and reducing the expressions of collagens and α-SMA in the kidney.

Actins ; analysis ; Animals ; Deoxyadenosines ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Protein-Serine-Threonine Kinases ; physiology ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; antagonists & inhibitors ; physiology

Recombinant human granulocyte-colony-stimulating factor (rhG-CSF) can widely regulate human immunologic response. In the protocol of peripheral blood stem/progenitor cell mobilization, rhG-CSF can change the numbers and functions of T cells. Then the results can impact the incidence of graft-versus-host disease after allogeneic peripheral blood stem/progenitor cell transplantation. The regulation of rhG-CSF on T cell is an indirect action which is based on the direct action to monocytes and dendritic cells. The numerous IL-10 secreted by monocytes plays a key role in cytokines production, proliferative response and cytotoxicity of T cells. Endogenous IL-10 can induce high expression of SOCS3 and the SOCS3 is very important for regulating the signal transduction of the activities of T cells. In this review influences of rhG-CSF on T-cells in mobilization process and related mechanisms were elaborated with emphasis.
Blood Donors ; Granulocyte Colony-Stimulating Factor ; pharmacology ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Interleukin-10 ; biosynthesis ; Recombinant Proteins ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; T-Lymphocytes ; cytology ; drug effects ; metabolism

LFA-1/ICAM-1 costimulation plays an important role in immunologic reaction of many different T cell populations. After TCR/CD3 complex cross-linking MHC/peptide, LFA-1, expressed on T cell increases a higher affinity and avidity for ICAM-1 rapidly. LFA-1 is a key molecule in formation of the immune synapse. LFA-1/ICAM-1 costimulation can engage various signaling events of T cell by up-regulating the activities of PI 3-kinase, sphingomyelinase, and c-Jun NH2-terminal kinase. With the costimulation of LFA-1/ICAM-1, engagement of TCR molecules results in a significant increase of T cell activities, including higher Th1 cytokines production, strongly proliferative response and higher T cell cytotoxicity.
Animals ; Cytokines ; biosynthesis ; Cytotoxicity, Immunologic ; Humans ; Intercellular Adhesion Molecule-1 ; physiology ; Lymphocyte Activation ; Lymphocyte Function-Associated Antigen-1 ; physiology ; Signal Transduction ; T-Lymphocytes ; immunology

Objective To investigate the effects of early rehabilitative training program on patients with acute myocardial infarction(AMI).Methods One hundred and twenty-two patients with AMI were randomly divided into early rehabilitation group(n=62)and control group(n=60).In addition to routine treatment,patients in rehabilitation group received early rehabilitative training mainly by walking exercise for two weeks.Results There were no significant differences in ventricular arrhythmia(Lown≥Ⅲ), extension of infarction and heart rate variability(HRV)between the two groups(P＞0.05).Forty of 62 patients(64.5%)in rehabilitation group had their left ventricular ejection fraction(LVEF)more than or equal to 50% in the 3~(rd)～4~(th)week after admission,significantly higher than that in control group(45.0%, 27/60 ;P＜0.01 ).By the end of the 4~(th)week after admission,25.8% of the patients in rehabilitation group showed positive in treadmill test,significantly lower than that in control group(38.3%,P＜0.01). Occurrence of angina pectoris and reinfarction and fatality in rehabilitation group were significantly lower than those in control group(P＜0.05)during their hospitalization and follow-up period.Patients in rehabilitation group stayed at hospital for(16?3)days in average,significantly less than that in control group[(27?4) days],with statistically significant difference(P＜0.05).Conclusion Early rehabilitative training for patients with uncomplicated AMI is not only safe and feasible,but also useful in improvement for their prognosis and quality of life.

Sphingosine 1-phosphate (S1P), a bioactive lipid produced by metabolism of sphingolipid, plays an important roles in the regulation of various biological responses. T cell expresses the S1P receptors, including S1P1, S1P2, S1P3, S1P4 and S1P5. Activation of S1P signal regulates multiple immunological functions of T cell, including proliferation, apoptosis, differentiation, migration and cytokine excretion. FTY720, a sphingosine analog, suppresses the S1P signal resulting in redistribution of lymphocytes from circulation to secondary lymphoid tissues, which has been applied as a potent immunosuppressive drug. In this paper, biosythesis and degradation of S1P, S1P receptor and its mediated signal pathway, S1P receptor expression of T-cells, regulation of S1P on T cell functions and immunosuppresion drugs involving S1P signal pathway were reviewed.
Fingolimod Hydrochloride ; Humans ; Immunosuppressive Agents ; pharmacology ; Lysophospholipids ; metabolism ; physiology ; Propylene Glycols ; pharmacology ; Receptors, Lysosphingolipid ; metabolism ; physiology ; Signal Transduction ; drug effects ; Sphingosine ; analogs & derivatives ; metabolism ; pharmacology ; physiology ; T-Lymphocytes ; drug effects ; immunology ; physiology

Objective To investigate the effect of L-arginine on expression of protein kinase C(PKC)mRNA during pulmonary ischemia and reperfusion injury(PIRI)in the rabbits.Methods Single lung ischemia and reperfusion animal model was used in vivo.The rabbits were randomly divided into three groups(n=9,in each),sham operated group (Sham),PIR group(I-R)and PIR+L-arginine group(L-Arg).Changes of several rariables including malondialdehyde (MDA),superoxide dismutase(SOD),malandialde hyde(MDA),nitril oxide(NO),wet to dry ratio of lung tissue weight(W/D)and index of quantitative assessment(IQA)of histolngic lung injury were recorded at 60 minutes after reperfusion in lung tissue.Meanwhile the location and expression of PKC mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60 minutes after reperfusion.Results In comparison with group I-R,PKC mRNA strongly expressed in intima and extima of small pulmonary artery as well as thin-waU vessels (mostly small pulmonary veins).The average optical density values of PKC-?,?and?mRNA in small pulmonary veins in L-Arg group had significance(all P＜0.01);SOD increased while MDA,W/D and IQA decreased at 60 minutes after reperfusion in lung tissue(P＜0.01 and P＜0.05).A morphologically abnormal changes of the lung tissue,were lessen markedly in L-Arg group.Conclusion L-arginine possess notably protective effects on PIRI in rabbits by activating PKC-?,?and?mRNA expression in lung tissue,raising NO level,dropping oxygen free radical level and decreasing lipid peroxidation.

OBJECTIVETo investigate the expression of nuclear factor kappa B (NF-kB), transforming growth factor beta1 (TGFbeta1), fibronectin (FN) in liver from diabetic rats.

METHODSTwenty male Sprague-Dawley rats were divided randomly into two groups: normal control group (n = 10) and type 2 diabetic group (n = 10). After 4 weeks of high-fat feeding, diabetic group rats were injected with low dosage streptozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat models. The diabetic rats received high-fat feeding for another 12 weeks. At the end of the experiment, the fibrosis lesion was observed under light microscopy after Masson staining. The mRNA levels of NF-kB, TGFbeta1, FN from rats liver were assayed by semi-quantity RT-PCR, the protein levels of NF-kB, TGFbeta1, FN was detected by IHC.

RESULTSFibrosis was found in diabetic rats. The levels of TGFbeta1, FN mRNA in liver tissues increased in diabetic rats compared with normal control rats (0.91+/-0.19 vs 0.47+/-0.20, t = 5.233, P less than 0.05; 1.85+/-0.70 vs 1.22+/-0.39, t = 2.463, P less than 0.05). And the protein levels of NF-kB P65, TGFbeta1, FN in liver tissues from diabetic rats were significantly higher than those in normal control rats (10978.77+/-8782.59 vs 4206.86+/-1430.56, Z = 1.979, P less than 0.05; 8551.00+/-4768.68 vs 4036.85+/-1051.12, Z = 2.303, P less than 0.05; 16980.30+/-11529.29 vs 5701.95+/-9461.75, t = -2.391, P less than 0.05).

CONCLUSIONUpregulation of NF-kB, TGFbeta1, FN in liver tissues may play a role in the hepatic fibrogenesis in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Fibronectins ; metabolism ; Liver ; pathology ; Liver Cirrhosis ; etiology ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. In order to investigate the role of sphingosine kinase-1 (SphK-1)/sphingosine 1-phosphate (S1P) signal pathway in the expression of CML cells, and to explore whether P210(bcr/abl) involved is activating SphK-1/S1P signal pathwey, the expressions of SphK-1 and S1P receptor mRNA in bcr/abl positive K562 cells and bcr/abl positive primary CML cells were detected by RT-PCR, the imatinib mesylate, the specific inhibitor of P210(bcr/abl) was employed to inhibit the P210(bcr/abl) tyrosine kinases of K562 cells and CML primary cells, and then the intracellular SphK-1 activity was assayed. The results indicated that after being cultured with 2.5 micromol/L imatinib mesylate for 0.5, 2, 6, 24 and 48 hours, the intensions of inhibiting SphK-1 activity were 0.007%, 38.9%, 34.6%, 28.1% and 76.1% resepectively. SphK-1 activity in CML cells also was reduced by 2.5 micromol/L imatinib mesylate (16.8% - 41.9% decrease). It is concluded that the CML cells express SphK-1 and different S1P receptor, and P210(bcr/abl) fusion protein in CML cells can activate SphK-1.
Benzamides ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Lysophospholipids ; genetics ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Signal Transduction ; genetics ; Sphingosine ; analogs & derivatives ; genetics ; metabolism

To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
ADP-ribosyl Cyclase ; immunology ; ADP-ribosyl Cyclase 1 ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Bone Marrow Cells ; cytology ; drug effects ; immunology ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Count ; Cell Division ; drug effects ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-1 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Time Factors

The study aimed to explore the changes of the immunocyte quantities and cytotoxicity in bone marrow, and how many hematopoietic progenitor cells retained after the bone marrow cells were activated by different combinations of various cytokines. Bone marrow cells were divided into four groups and were cultured in vitro: (1) Control: no cytokines were added. (2) IL-2 group: bone marrow cells were activated by IL-2. (3) CD3-AK group: bone marrow cells were activated by IL-2 and CD3-McAb. (4) CIK group: IFN-gamma, IL-1, IL-2 and CD3-McAb were added. CFU-GM assay and CD3 phenotype detection were performed before and after activating culture in all groups. The changes of cell quantities during culture and cytotoxicity of cultured cells were tested. CD3 positive cells markedly increased in both CD3-AK and CIK groups. The cell numbers and cytotoxicity of CD3-AK and CIK groups were higher than those of control or IL-2 group obviously after culture (P < 0.05). CFU-GM were decreased in all groups after culture and there had no significant difference among four groups. The combination of IL-2 and CD3-McAb not only stimulates the proliferation of marrow immunocytes and increases their cytotoxicity but retains enough hematopoietic progenitor cells as well. This combination of cytokines can be used to purge autologous bone marrow in vitro.