Astragalus membranaceus may be a potential therapy for childhood asthma but its driving mechanism remains elusive. The main components of A. membranaceus were identified by HPLC. The children with asthma remission were divided into two combination group (control group, the combination of budesonide and terbutaline) and A. membranaceus group (treatment group, the combination of budesonide, terbutaline and A. membranaceus). The therapeutic results were compared between two groups after 3-month therapy. Porcine peripheral blood mononuclear cells (PBMCs) were isolated from venous blood by using density gradient centrifugation on percoll. The levels of FoxP3, EGF-β, IL-17 and IL-23 from PBMCs and serum IgE were measured. The relative percentage of Treg/Th17 cells was determined using flow cytometry. The main components of A. membranaceus were calycosin-7-O-glucoside, isoquercitrin, ononin, calycosin, quercetin, genistein, kaempferol, isorhamnetin and formononetin, all of which may contribute to asthma therapy. Lung function was significantly improved in the treatment group when compared with a control group (P < 0.05). The efficacy in preventing the occurrence of childhood asthma was higher in the treatment group than the control group (P < 0.05). The levels of IgE, IL-17 and IL-23 were reduced significantly in the treatment group when compared with the control group, while the levels of FoxP3 and TGF-β were increased in the treatment group when compared with the control group (P < 0.05). A. membranaceus increased the percentage of Treg cells and reduced the percentage of Th17 cells. A. membranaceus is potential natural product for improving the therapeutic efficacy of combination therapy of budesonide and terbutaline for the children with asthma remission by modulating the balance of Treg/Th17 cells.
Animals ; Asthma ; drug therapy ; immunology ; Astragalus propinquus ; chemistry ; Budesonide ; administration & dosage ; Cells, Cultured ; Child ; Child, Preschool ; Cytokines ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Humans ; Immunologic Factors ; administration & dosage ; pharmacology ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Lung ; drug effects ; physiology ; Male ; Swine ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; Terbutaline ; administration & dosage ; Th17 Cells ; cytology ; drug effects ; Treatment Outcome

This study is to investigate the anti-tumor effect in vitro of methotrexate modified by LH-RH peptide (LH-RH-MTX). LH-RH receptors highly expressing MCF-7 human breast carcinoma cell line and lowly expressing K562 human erythroleukemia cell line were served as the tested cells. The cell proliferation inhibition rates of LH-RH-MTX were detected by MTT colorimetric assay. The effects of LH-RH-MTX on the cell cycle and apoptosis rates were detected by flow cytometry. The inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that on K562 cells, and the inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that of free MTX at the same concentration. The inhibition rate of LH-RH-MTX on rat bone marrow mononuclear cells was less than that of free MTX. The number of MCF-7 cells in S phase increased after administration of LH-RH-MTX. The apoptosis rate of LH-RH-MTX group significantly increased compared with that of the control group and MTX group. The relative expression of LHRHR mRNA of LH-RH-MTX group markedly decreased compared with that of the control group and MTX group. LH-RH-MTX is realizable to reduce drug side effects, increase the therapeutic index and achieve tumor-targeted therapy.
Animals ; Antimetabolites, Antineoplastic ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Delivery Systems ; Gonadotropin-Releasing Hormone ; chemistry ; pharmacology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; MCF-7 Cells ; Methotrexate ; chemical synthesis ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptors, LHRH ; biosynthesis ; genetics

The purpose of this study is to find out anti-HIV-1 reverse transcriptase (RT)/protease (PR) activity and inhibition of virus replication in cell cultures of novel coumarin analogs and determine their structure-activity relationship. Coumarin derivatives have been demonstrated to inhibit the activity of HIV-1 RT/PR in cell free system. It also shows inhibition effects to HIV-1 replication in cell culture. Based on the Chinese traditional pharmacological characteristics and protein three dimension computer aided design, analogs of tetracyclic dipyranocoumarin were synthesized from natural leading compounds. We studied the relationship of antiviral effects and chemical structures via HIV-1 PR/RT enzyme models and cell culture model system. Seven compounds were designed and tested. Several compounds showed anti-HIV-1 activity in varying degrees, especially V0201 showed much higher anti-HIV-1 activity with 3.56 and 0.78 micromol x L(-1) of IC50 against HIV-1 PR/RT and 0.036 micromol x L(-1) against HIV-1 replication in PBMC cultures. V0201 with a novel structure may be a new leading compound. These new compounds are valuable for development of new anti-HIV drugs in the future.
Anti-HIV Agents ; chemical synthesis ; chemistry ; pharmacology ; Cells, Cultured ; HIV Core Protein p24 ; metabolism ; HIV Protease ; metabolism ; HIV Reverse Transcriptase ; metabolism ; HIV-1 ; physiology ; Humans ; Inhibitory Concentration 50 ; Leukocytes, Mononuclear ; cytology ; metabolism ; virology ; Pyranocoumarins ; chemical synthesis ; chemistry ; pharmacology ; Reverse Transcriptase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Virus Replication ; drug effects

BACKGROUNDPrevious studies showed that anti MHC-II monoclone antibody (MAb) only had partial inhibiting effect of alloreactive mixed lymphocyte reaction (MLR) in vitro and it was unsteady and non-persistent. The aim of this research was to determine whether radioactive isotope (188)Re marked MHC-II antibody could benefit the allograft acceptance in transplantation as compared to normal MHC-II antibody.

METHODS188Re was incorporated to 2E9/13F (ab')(2) which is against swine MHC class II antigen (MAb-(188)Re). Porcine peripheral blood mononuclear (PBMC) cells were examined for proliferation and cytokine mRNA expression after stimulation with MHC-II MAb or MAb-(188)Re.

RESULTSThe proliferative response of recipient PBMCs in mixed lymphocyte reaction (MLR) to donor alloantigen showed that the stimulation index of MAb-(188)Re group was significantly lower than the MHC-II MAb group and control (P < 0.05). mRNA expression of interleukin 2, interferon Υ and tumor necrosis factor α (type 1 cytokines) was lower in MAb-(188)Re group than the MHC-II MAb group, while interleukin 10 (type 2 cytokines) was higher in MAb-(188)Re group in the first 24 hours.

CONCLUSIONMAb-(188)Re could help the graft acceptance by inhibiting T cell proliferation, lowering the expression of type 1 cytokines and elevating the type 2 cytokines produced by PBMC.

Animals ; Antibodies, Monoclonal ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Interleukin-10 ; genetics ; Interleukin-2 ; genetics ; Isoantigens ; immunology ; Leukocytes, Mononuclear ; drug effects ; radiation effects ; Lymphocyte Culture Test, Mixed ; Mitomycin ; pharmacology ; Radioisotopes ; Reverse Transcriptase Polymerase Chain Reaction ; Rhenium ; Swine ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo study the effects of TNF-α on ICAM-1 and LFA-1 expression in peripheral blood mononuclear cells (PBMC) of children with febrile seizures (FS).

METHODSSixteen children with FS and 16 age- and gender-matched healthy children were enrolled. The samples of PBMC from FS children were randomized into two groups with or without TNF-α treatment (TNF-α concentration 1.0 ng/mL). PBMC were purified and cultured with a conventional method in vitro. The expression of ICAM-1 and LFA-1 in PBMC was determined by flow cytometry (FCM).

RESULTSICAM-1［(20±9)% vs (14±7)%)］and LFA-1［(43±16)% vs (30±16)%］expression in PBMC in the untreated FS group was significantly higher than that in the normal control group (P<0.05). Compared with the untreated FS group, the treatment with TNF-α remarkably increased the ICAM-1 expression［(27±11)%］(P<0.05). PBMC LFA-1 expression［(52±21)%］in the TNF-α-treated group was higher than that in the untreated FS group, although there were no statistical differences between the two groups.

CONCLUSIONSTNF-α treatment may increase LFA-1 and ICAM-1 expression in PBMC of children with FS.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Intercellular Adhesion Molecule-1 ; blood ; Leukocytes, Mononuclear ; chemistry ; drug effects ; Lymphocyte Function-Associated Antigen-1 ; blood ; Male ; Seizures, Febrile ; immunology ; Tumor Necrosis Factor-alpha ; pharmacology

BACKGROUNDIt is well known that dehydration can impair bodily functions. To evaluate the impact of hydration status under ambient environmental temperature on the immune system, 25 male collegiate wrestlers were recruited to undergo an experimental dehydration program.

METHODSThirteen subjects had controlled diets with individual energy requirements to prevent body mass loss and restricted water intake to cause 4.52% dehydration; they formed the dehydrated group (DE). These subjects developed a urine specific gravity of about 1.030 in 84 hours. Twelve other subjects had no water restriction and maintained their total body weight comprised the euhydrated group (EU). Peripheral blood monocytes (PBMNC) were isolated after dehydration to perform immune response testing by being incubated with a polysaccharide fraction from fu-ling, Poria cocos (polysaccharide fraction from Poria cocos, PCPS, 1 - 30 £g/L), to prepare a conditioned medium termed conditioned medium of PBMNC stimulated by PCPS (PCPS-MNC-CM). More PCPS (25 µg/L) was needed in the DE group to prepare the PCPS-MNC-CM, which was assayed with a growth inhibitory curve for treated U973 cells.

RESULTSThe treated U937 cells, incubated together with PCPS-MNC-CM from the DE group, exhibited a much lower nitroblue tetrazolium (NBT) positive value of (63.7 ± 4.7)%. The concentration of interferon-gamma (IFN-γ), interleukin (IL)-1β and tumor necrosis factor (TNF)-α in PCPS-MNC-CM from subjects after dehydration was much lower than in the CM from the EU group.

CONCLUSIONThe immune response to PCPS in the DE group was lower than in normally hydrated subjects.

Adolescent ; Adult ; Dehydration ; physiopathology ; Drugs, Chinese Herbal ; chemistry ; Fever ; Humans ; Immunologic Factors ; Interferon-gamma ; metabolism ; Interleukin-1beta ; metabolism ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Polysaccharides ; chemistry ; immunology ; Tumor Necrosis Factor-alpha ; metabolism ; U937 Cells ; Universities ; Wolfiporia ; Wrestling ; Young Adult

Resveratrol, isorhapontigenin and pinosylvin, isolated from Gnetum parvifolium, and their analogues have been synthesized and tested for their inhibitory activity of HIV-1. Natural product 12a and analogues (12d, 12e, 12g) display significant inhibitory activity of HIV-1 replication. Among them, compound 12d (trans-3, 4, 5, 4'-tetrahydroxystilbene) exhibits the most potent anti-HIV-1 activity with an IC50 value of 1.84 micromol x L(-1).
Anti-HIV Agents ; isolation & purification ; pharmacology ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gnetum ; chemistry ; HIV-1 ; drug effects ; physiology ; Inhibitory Concentration 50 ; Leukocytes, Mononuclear ; cytology ; virology ; Molecular Structure ; Plants, Medicinal ; chemistry ; Stilbenes ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo study the mobilization efficiency effect on bone marrow stem cells by Panax notoginseng on acute myocardial infarction in experimental rats.

METHODOne hundred and thirty wistar clean rats with average age 16 weeks weighing (200 +/- 10) g, half female, ligated anterior descending coronary artery proximal,were established animal model of acute myocardial infarction. They were randomly divided into: model group, Chinese medicine mobilization group, western medicine mobilization group, integrated Chinese and western medicine mobilization group and sham group. Except sham group rats, other groups after the survival of 24 h by 1, 7, 14 d, and other points in time were divided into three subgroups. They were injected drugs immediately 24 h after the surgery. Injected G-CSF 50 ug x kg(-1) x d(-1) in western medicine group subcutaneously, NS 50 g x kg(-1) x d(-1) in sham group and model group. In Chinese medicine mobilization group intraperitoneally injected notoginsenoside (150 mg x kg(-1)). Chinese and western medicines mobilization group were subcutaneously injected G-CSF 5 microg x kg(-1) x d(-1) and intraperitoneally injected notoginsenoside (150 mg x kg(-1)), with G-CSF once a day, 7 days totally and G-CSF once a day until each sub-group of animals were killed. Severn rats were remained for experiments in every subgroup. Flow cytometry were used to detect the percentage of peripheral blood mononuclear cells and white blood, C-kit receptor expression of peripheral blood mononuclear cells.

RESULTWBC, MNC% of model group 1, 7, 14 d were significantly larger than those of sham group 1, 7, 14 d (P < 0.05), which indicates that MI animals can mobilize themselves. WBC and MNC% of Chinese medicine group and western medicine group were lower than those of Chinese and western medicine group in 1, 7, 14 d (P < 0.05). C-kit receptor expression in PBMC of Chinese and western medicine group and western medicine group were higher than those of model group. Western medicine is better than Chinese medicine in mobilization.

CONCLUSION: Panax notoginseng may cooperation G-CSF with mobilizing stem cell, raise the efferens efficiency of bone marrow stem cells, increase the number of peripheral blood stem cells.

Animals ; Bone Marrow Cells ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Hematopoietic Stem Cell Mobilization ; Leukocyte Count ; Leukocytes, Mononuclear ; drug effects ; Male ; Myocardial Infarction ; blood ; metabolism ; pathology ; Panax notoginseng ; chemistry ; Rats ; Rats, Wistar ; Stem Cells ; cytology ; drug effects ; Time Factors

OBJECTIVETo evaluate the effect of fumigating with Yinxieling (YXL) in treating patients with psoriasis vulgaris and its influence on T-bet and GATA-3 protein expressions in peripheral blood monocyte (PBMC).

METHODSWestern blot method was employed to detect the T-bet and GATA-3 protein expressions in PBMC of 30 psoriasis vulgaris patients before and after they received fumigation therapy with YXL, also in 25 healthy persons for controls. The therapeutic efficacy was observed and the relationship between PASI scores and levels of T-bet and GATA-3 analyzed.

RESULTSAfter treatment, 12 out of the 30 patients were cured, 9 were markedly effective, 8 effective and 1 unchanged, the cure rate being 40.0% and the effective rate 96.7%. Level of T-bet expression in PBMC of patients was 1.7917 +/- 0.3840, which was higher than that of healthy persons (0.8860 +/- 0.1486, P < 0.01), but the GATA3 expression was lower than that in control (0.8777 +/- 0.3114 vs. 1.2384 +/- 0.1783, P < 0.01). However, the two indexes were restored after fumigation to 1.3410 +/- 0.3642 and 1.0883 +/- 0.2435 respectively, showing significant difference to those before fumigation (P < 0.01). Correlation analysis showed that PASI score was positively correlated with level of T-bet expression (r = 0.7448, P < 0.01) and negatively correlated with level of GATA-3 expression (r = -0.8291, P < 0.01).

CONCLUSIONFumigation therapy is effective in treating psoriasis vulgaris, its mechanism is possibly by way of modulating the equilibrium of the transcription factors T-bet and GATA-3 protein expressions in PBMC, and rectifying the immune abnormality of Th1/Th2 subsets imbalance.

Administration, Inhalation ; Adolescent ; Adult ; Animals ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; GATA3 Transcription Factor ; genetics ; immunology ; Gene Expression ; drug effects ; Humans ; Leukocytes, Mononuclear ; drug effects ; immunology ; Male ; Middle Aged ; Psoriasis ; drug therapy ; genetics ; immunology ; T-Box Domain Proteins ; genetics ; immunology ; Young Adult

OBJECTIVETo explore and evaluate the activities of composite extract from Salvia Yunnanensis and in cell cultures (DS-MEF) for inhibition of human immuno-deficiency virus type 1 (HIV-1) in vitro and in cell cultures.

METHODSThe inhibitory activity of DS-MEF on HIV-1 reverse transcriptase (RT), protease (PR) and integrase (IN) were detected in vitro with radionuclide 3H incorporation, fluorescence assay and enzyme-linked immunosorbent assay respectively. The human T-lymphocyte MT-4 cell line, human T-lymphocyte H 9 cell line chronically infected with HIV-1 IIIB, and the fresh peripheral blood mononuclear cell (PBMC) of healthy persons as well as the laboratory passed HIV-1 IIIB and the clinically isolated HIV-1 AZT sensitive 018a or resistant 018c infected cell cultures were used for evaluating the cytotoxicities and inhibitory activities of DS-MEF on HIV-1 P 24 antigen. The acute toxicities of DS-MEF on KM mice were determined by gastric gavages and intraperitoneal injections with various dosages.

RESULTSThe IC50 of DS-MEF for inhibiting HIV-1 IN, RT and PR were 2.59 +/- 0.50 mg/L, 27.39 +/- 11.18 mg/L and 9.38 +/- 2.45 mg/L respectively. In MT-4 cell cultures infected with HIV-1 III, TC50 were 13.19 +/- 6.07 mg/L, IC50 and SI of anti-HIV-1 activity were 0.224 +/- 0.163 mg/L and 58.7; in chronically infected H 9 cell cultures, TC50 were 18.11 +/- 9.84 mg/L, IC50 on HIV-1 P 24 antigen and SI were l7.230 +/- 21.114 mg/L and 1.1 respectively; TC50 in HIV-1 infected PBMC cultures were 288.70 +/- 0.08 mg/L; IC50 on AZT sensitive HIV-1 018a: 26.42 +/- 11.16 mg/L, and SI: 10.9; On AZT resistant HIV-1 018c, IC50: 27.87 +/-5.35 mg/L, and SI: 10.4. Moreover, DS-MEF showed synergistic effect with AZT or nevirapine (NVP) on HIV-1 IIIB in MT-4 cell cultures, the respective combination index was 0.78 or 0.67. DS-MEF showed no acute toxicity in KM mice with the dosage up to 20 g/kg via gastrogavage, and the 50% lethal dose (LD50) via intraperitoneal injection was 1.18 g/kg.

CONCLUSIONDS-MEF is a promising anti- HIV-1 agent with low toxicity in mice and possesses multi-targets and effective activities.

Animals ; Anti-HIV Agents ; pharmacology ; therapeutic use ; Cell Line ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; HIV Infections ; drug therapy ; virology ; HIV-1 ; drug effects ; enzymology ; physiology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Salvia ; chemistry ; T-Lymphocytes ; virology ; Viral Proteins ; antagonists & inhibitors ; Virus Replication ; drug effects