Objective To explore the protective mechanism of honey-processed Hedysari Radix in regulating intestinal mucosal injury in rats with spleen qi deficiency.Methods The three-factor composite modeling method of bitter cold diarrhea,overwork and hunger and satiety disorder was used to construct a spleen qi deficiency model rats.After the model was successfully made,they were randomly divided into model group,honey-processed Hedysari Radix group and probiotic group,with 15 animals in each group.Another 15 normal rats were taken as the blank group.The honey-processed Hedysari Radix group was given 12.6 g·kg-1 water decoction of honey-processed Hedysari Radix by gavage,the probiotics group was given Bifidobacterium Lactobacillus triple viable tablets suspension at a dose of 0.625 g·kg-1,and the blank group and the model group were given the same dose of distilled water.The rats in the four groups were administered once a day for 15 days.Enzyme-linked immunosorbent assay was used to detect diamine oxidase(DAO)in serum,D-lactic acid(D-LA),secretory immunoglobulin A factor,and Western blotting was used to detect the expression levels of AMP-activated protein kinase(AMPK),zonula occludens-1(ZO-1)and occludin in colon tissues.Results The serum levels of DAO in the blank group,model group,honey-processed Hedysari Radix group and probiotic group were(138.93±9.78),(187.95±12.90),(147.21±6.92)and(166.47±3.37)pg·mL-1;the contents of D-LA were(892.23±49.17),(1 099.84±137.64),(956.56±86.04)and(989.61±51.75)μg·L-1;the contents of SIgA in colon tissues were(14.04±1.42),(11.47±2.39),(11.84±1.49)and(12.93±1.65)μg·mL-1;the relative expression levels of ZO-1 protein in colon tissues were 1.18±0.11,0.42±0.04,0.77±0.05 and 0.95±0.07;the relative expression levels of occludin protein were 1.35±0.31,0.61±0.17,1.19±0.19 and 0.88±0.13;the relative expression levels of AMPK protein were 0.91±0.02,0.35±0.09,0.74±0.08 and 0.59±0.11.Compared with the model group,there were significant differences in the serum content of DAO and D-LA,SIgA content in colon,and the content of ZO-1,occludin and AMPK protein in the honey-processed Hedysari Radix group(P＜0.01,P＜0.05).Conclusion Honey-processed Hedysari Radix can enhance the protective effect on the intestinal mucosa of rats with spleen qi deficiency by regulating the expression of related inflammatory cytokines,intestinal mucosal upper cell enzymes and tight junction proteins in rats with spleen qi deficiency.

Objective:To study the effects of Banxia Baizhu Tianmatang （BBTT） on atherosclerosis in apolipoprotein-E knockout （ApoE^-/-） mice induced by high fat diet. Method:The atherosclerosis model of ApoE^-/- mice was established with high-fat diet， and BBTT was used for intervention. The pathological changes of aorta after atherosclerosis were observed by hematoxylin-eosin （HE）， oil red O and Masson staining. The changes of serum total cholesterol （TC）， triglyceride （TG）， high density lipoprotein cholesterol （HDL-C） and low density lipoprotein cholesterol （LDL-C） were detected by automatic biochemical analyzer. The expression levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） and oxidized low density lipoprotein （ox-LDL） were detected by enzyme linked immunosorbent assay （ELISA）. Total tissue proteins were extracted， quantified by protein quantification （BCA） method， and the expression of matrix metalloproteinase-9 （MMP-9） protein was detected by Western blot. Thiobarbituric acid （TBA） method was used to detect the change of malondialdehyde （MDA） content. The change of superoxide dismutase （SOD） activity was detected by 2-（2-methoxy-4-nitrophenyl）-3-（4-nitrophenyl）-5-（2，4-disulfobenzene）-2H tetrazole monosodium salt （WST-8） method. Result:Compared with the control group， there was a large amount of lipid accumulation in the blood vessels of the model group， the serum levels of TG， TC， HDL-C and LDL-C significantly increased （P<0.05）， the expression of MMP-9 protein in the blood vessels significantly increased （P<0.01）， the expression levels of IL-6 and TNF-α in the serum increased （P<0.05， P<0.01）， the SOD activity was significantly reduced （P<0.01）， and the levels of MDA and ox-LDL expression increased （P<0.01）. Compared with the model group， the treatment with BBTT could inhibit the accumulation of lipids in blood vessels， the TG levels were reduced in the high and medium dose groups of BBTT （P<0.05）， high， medium and low dose groups significantly reduced the levels of LDL-C in serum （P<0.01）， the expression of MMP-9 protein in blood vessels （P<0.05） and IL-6 in serum （P<0.01）， the high-dose group down-regulated the expression of TNF-α in serum （P<0.01） and ox-LDL （P<0.01）， both the high and medium-dose groups increased the level of MDA （P<0.05， P<0.01） and the activity of SOD （P<0.05）. Conclusion:BBTT has a certain intervention effect on the formation of atherosclerosis aortic plaque in ApoE^-/- mice， and its mechanism may be associated with reducing the TG and LDL-C levels， lowering blood lipid， down-regulating MMP-9 protein， protecting blood vessels from inflammatory damage， reducing ox-LDL and MDA levels， and improving SOD activity to play an antioxidant role.

Hematopoietic Stem Cell Transplantation ; Humans ; Nocardia Infections ; Transplantation, Homologous

Analgesics, Opioid ; chemistry ; pharmacology ; Animals ; Cattle ; Diazepam ; chemistry ; pharmacology ; Diazepam Binding Inhibitor ; chemical synthesis ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Mice ; Morphine ; chemistry ; pharmacology ; Receptors, GABA-A ; metabolism ; Swine

OBJECTIVETo investigate and analyze the traumatic conditions of Longnan area of Gansu in Wenchuan earthquake, and to explore the measures coping with unexpected disasters.

METHODSA sample survey including questionnaire survey, interviews and other methods was adopted to investigate the conditions of affected population of Longnan area in earthquake, especially on the traumatic conditions, the injured causes, treatment methods.

RESULTSIn the sample survey of a total of 1652 earthquake casualties, the dead were 16 and the patients with extremities fractures were 750 (45.4%). Organized and timely rescue of the wounded could reduce the complications and the mortality .

CONCLUSIONStrengthening the prevention and treatment of earthquake trauma can effectively decrease the death rate and disabled rate.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Data Collection ; Disasters ; statistics & numerical data ; Earthquakes ; mortality ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Wounds and Injuries ; mortality ; psychology ; surgery ; therapy

OBJECTIVETo analyze the expression of NKG2D ligands on human nasopharyngeal carcinoma cell line CNE2 and the multidrug-resistant lin CNE2/DDP and investigate its impact on cytotoxicity of natural killer (NK) cells.

METHODSExpression of NKG2D ligands on the surface of CNE2 and CNE2/DDP cells was analyzed by flow cytometry, and their HLA genotypes, along with inhibitory killer cell immunoglobulin-like receptors (KIRs) expressed on NK cells from 5 healthy donors, were determined by PCR with sequence specific primers. Cytotoxicity of NK cells against CNE2 and CNE2/DDP cells was evaluated by LDH-releasing assay at different effector-to-target ratios (E:T). In blocking experiments, different monoclonal antibodies (mAb) were added to the target cells at the E:T of 20:1 ratio.

RESULTSThe HLA genotypes of CNE2 and CNE2/DDP cells were A2, 24, B18, 35, Cw4, 7, and the inhibitory KIR genotypes of the 5 healthy donors were KIR2DL1, KIR2DL3, KIR3DL1, and KIR3DL2, mismatched with the HLA -class I molecules expressed by the CNE2 and CNE2/DDP cells. The expression of MHC class I chain-related proteins A and B (MICA and MICB) on CNE2 cells was higher than that on CNE2/DDP cells (P<0.01), and ULBP1 and ULBP3 were not detectable. NK cells displayed highly in vitro cytotoxicity against CNE2 and CNE2/DDP cells with a mean cell lysis rate of (10.50-/+2.17)%, (4.98-/+0.95)%; (27.68-/+1.47) %, (15.48-/+2.10) %; (36.99-/+3.13) %, (28.46-/+4.30) %; (55.00-/+2.20) %, (40.95-/+2.21) %, respectively, corresponding to the E:T ratios of 5:1, 10:1, 20:1, and 30:1 (P<0.01). Blocking experiments confirmed that killing of CNE2 and CNE2/DDP cells by NK cells was efficiently inhibited by anti-MICA, anti-MICB, and anti-ULBP2 mAbs, whereas anti-ULBP1 and anti-ULBP3 mAbs had no effects on the cytotoxicity of the NK cells.

CONCLUSIONExpression of NKG2D ligands on CNE2 and CNE2/DDP cells is correlated with NK cell-mediated lysis, and NK cells display higher cytotoxicity against parental CNE2 cells than the multidrug-resistant CNE2/DDP cells.

Antibodies ; pharmacology ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; drug effects ; immunology ; Drug Resistance, Multiple ; Flow Cytometry ; Genotype ; HLA Antigens ; genetics ; Histocompatibility Antigens Class I ; immunology ; metabolism ; Humans ; Killer Cells, Natural ; immunology ; NK Cell Lectin-Like Receptor Subfamily K ; immunology ; metabolism ; Nasopharyngeal Neoplasms ; immunology ; metabolism ; pathology ; Polymerase Chain Reaction ; Receptors, KIR ; genetics

In order to investigate the central nervous mechanism and the diseases involved in catecholamine transmitter secretion, the dynamics of catecholamine release is studied in single cell, brain slice or in vivo. Noradrenaline is an important neurotransmitter and modulator in the central nervous system (CNS) and the peripheral nervous system (PNS). In the present paper, we first compared three real-time methods used to measure noradrenaline secretion in single cells (membrane capacitance, amperometry and confocal fluorescence microscopy imaging). Compared to the electrophysiological method and fluorescence microscopy, the basic usage of the carbon fiber electrode (CFE) in neuroscience research was presented as an example. Then, we presented a primary description of ion channels, including voltage-gated Na(+), K(+) and Ca(2+) channels in locus coeruleus (LC) neurons in rat brain slices. Finally, we presented example recordings of combined patch-clamp and amperometry measurements in LC neurons, indicating Ca(2+)-dependent quantal noradrenaline release following Ca(2+) influx through Ca(2+) channels.
Animals ; Central Nervous System ; physiology ; Ion Channels ; physiology ; Norepinephrine ; secretion ; Patch-Clamp Techniques ; Rats

OBJECTIVETo observe the effect of thermotherapy on the intracellular adriamycin concentration and apoptosis of Raji cells in vitro.

METHODSThe working concentration of adriamycin against Raji cells was determined with MTT assay. Raji cells were subjected to thermotherapy (at 40 degrees Celsius;, 41 degrees Celsius; or 42 degrees Celsius;) and chemotherapy with adriamycin alone or in conjunction, and the cell survival rates were determined 48 h after the treatment. The cell growth inhibition effect of the treatment was evaluated with MTT assay, and the apoptotic rates of Raji cells and alteration of intracellular adriamycin concentration were determined by flow cytometry.

RESULTSThe IC(50) of adriamycin was defined as the working concentration in the experiment. Thermotherapy at 40, 41 and 42 degrees Celsius; for 60 min in conjuction with chemotherapy significantly inhibited the growth of Raji cells (P<0.01). The results of flow cytometry showed that thermotherapy and adriamycin chemotherapy, used either alone or in combination, significantly increased the apoptotic rates of Raji cells (P<0.01), and thermotherapy remarkably increased the intracellular concentration of adriamycin.

CONCLUSIONAdriamycin chemotherapy combined thermotherapy for 60 min can increase the intracellular concentration of adriamycin and the apoptosis rates of Raji cells.

Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Combined Modality Therapy ; Doxorubicin ; metabolism ; pharmacology ; Flow Cytometry ; Hot Temperature ; Humans ; Hyperthermia, Induced ; Inhibitory Concentration 50 ; Intracellular Space ; metabolism ; Lymphoma ; drug therapy ; metabolism ; pathology

This study was purposed to investigate the inhibitory effect, apoptosis, Bcl-2 and P-gp expression of K562/AO2 cells by hyperthermia combined with adriamycin. The working concentration of adriamycin against K562/AO2 was determined by MTT assay. The hyperthermia and chemotherapy were used alone or in combination, then the cell survival rate was detected at 48 hours. The inhibitory effect was evaluated by MTT assay. The apoptosis rate, Bcl-2 and P-gp expression of K562/AO2 were determined by flow cytometry. The concentration of adriamycin in the experiment was defined as its IC(50) at 48 hours action. The results indicated that the hyperthermia at 40, 41 and 42 degrees C for 60 minutes showed obvious inhibitory effect on K562/AO2 cells (p < 0.01). Adriamycin chemotherapy combined with hyperthermia showed more obvious inhibitory effect on K562/AO2. According to flow cytometric analysis, the hyperthermia and adriamycin used alone or in combination could obviously increase the apoptosis rate and down-regulate Bcl-2 and P-gp expression of K562/AO2 cells (p < 0.01). It is concluded that the adriamycin chemotherapy combined with hyperthermia for 60 minutes shows obvious inhibitory effect on K562/AO2 cells, which increases the apoptosis rate and down-regulates expression of Bcl-2 and P-gp.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Neoplastic ; Humans ; Hyperthermia, Induced ; K562 Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

Objective To investigate the inhibitory effect of adriamycin in combination with hyperthermia on apoptosis and bcl-2 expression in the chronic leukemic cell line K562/AO2 in vitro.Methods The working con- centration of adriamycin against K562/AO2 determined by using methyl thiazolyl tetrazolium(MTT) assay was used to treat the chronic leukemic cell line K562/AO2 in vitro alone or in combination with hyperthermia induced using a hot water bath at 40,41 or 42℃.The inhibitory effect was evaluated by MTT assay.The apoptosis rates and bcl-2 ex- pression of K562/AO2 were determined by flow cytometry.Results The working concentration of adriamycin in the study was defined as its 50% inhibition concentration (IC50).A 60 min session of hyperthermia at 40℃,41℃or 42℃was associated with significant growth inhibition of the cell line K562/AO2.Adriamycin chemotherapy alone and with hyperthermia significantly inhibited the growth of K562/AO2.All treatments significantly increased apoptosis rates and down-regulated bcl-2 expression of the K562/AO2 cell line.Conclusion Adriamycin chemotherapy com- bined with 60 min sessions of hyperthermia showed significant suppression effect on K562/AO2 cell proliferation.The treatment can increase apoptosis rates and down-regulate bcl-2 expression.