In order to clarify material basis of effective parts of liangxue tongyu prescription, blood-heat and blood-stasis rat model induced by dry yeast was established. The changes of rectal temperature, blood viscosity and plasma viscosity were used to evaluate the cooling-blood and activating-blood effects of liangxue tongyu prescription and its parts. Compared with the model group, the extract from liangxue tongyu prescription, its volatile oil and n-butanol part could significantly reduce rectal temperature (P<0.01), and also reduce blood viscosity and plasma viscosity to various degrees (P<0.01 or P<0.05). So volatile oil and n-butanol part were primarily identified as effective parts of liangxue tongyu prescription. By using GC-MS with normalization method of area to analyze volatile oil of liangxue tongyu prescription, 70 compounds were identified, accounting for about 92.54%, mainly as β-asarone, paeonol, α-asarone and shyobunone. 42 compounds such as peony glycosides, tannins, and iridoid glycosides were identified by HPLC-MS techniques and standard comparison. The study determined the effective parts of liangxue tongyu prescription and clarified the chemical composition providing the foundation for further studies on material basis of liangxue tongyu prescription.
Acetophenones ; chemistry ; Animals ; Anisoles ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; chemistry ; Rats ; Tannins ; chemistry

OBJECTIVETo elucidate the gene regulatory pattern of Epimedium flavonoids (EF) in immune homeostasis remodeling in the aged rats.

METHODS(1) To quantitatively analyse the apoptosis percentage of lymphocyte in spleen of aged, young and EF treated rats using flow cytometry. (2) To analyse the lymphocyte gene expression profiles of different groups using gene chips (Rat Genome U34A).

RESULTS(1) Detection of lymphocyte apoptosis percentage showed that there was significant difference in comparing between aged group and young group, between EF treated group and aged group (P < 0.01). (2) As compared with that in the young group, in the aged group, 116 genes were up-regulated and 215 down-regulated. As compared with that in the old group, in the EF treated group, 447 genes were up-regulated and 456 down-regulated, which involved the aspects as cell apoptosis and cell proliferation regulation, etc.

CONCLUSION(1) The expression pattern characterized by up-regulation of apoptosis promoting genes expression and down-regulation of apoptosis inhibiting genes expression, is the important gene background of immuno-homeostasis imbalance in the aged. (2) The role of EF is to reverse the abnormal changes of gene expressions with opposite functions, i.e. the apoptosis promoting and inhibiting, proliferation enhancing and antagonizing genes, to reconstruct a beneficial equilibrium of gene expression and thus to further remodel the immuno-homeostasis in the aged.

Adjuvants, Immunologic ; pharmacology ; Aging ; Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Female ; Flavonoids ; pharmacology ; Gene Expression ; Gene Expression Profiling ; Homeostasis ; Lymphocytes ; immunology ; metabolism ; Male ; Neuroimmunomodulation ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo compare the modulating effect of Kidney tonifying recipe (KTR) and Spleen invigorating recipe (SIR) on T-lymphocyte apoptosis (TLA) in corticosterone treated rats (CTR).

METHODSQualitative and quantitative analysis on activation-induced TLA was conducted by transmission electron microscope and TUNEL labelled flow cytometry detection.

RESULTSTLA percentage of CTR was 45.87 +/- 7.22%, while that of normal control rats was 34.25 +/- 6.47%, the difference between them was significant (P < 0.01). TLA percentage of Youguiyin treated rats was 35.90 +/- 7.39%, and that of Bushen Yishou capsule treated rats was 36.20 +/- 9.14%, compared with that of CTR, P < 0.01 and P < 0.05 respectively. TLA percentage of the SIR treated group was 36.92 +/- 11.82%, which was different insignificantly as compared with that of CTR.

CONCLUSIONTLA susceptibility was significantly enhanced in CTR. Both KTRs (Youguiyin and Bushen Yishou capsule) had down-regulating effect on TLA, while the effect of SIR was insignificant, suggesting that down-regulating activation-induced TLA may be one of the important mechanisms of KTR in improving T-lymphocyte function in CTR.

Animals ; Apoptosis ; drug effects ; Corticosterone ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; cytology ; ultrastructure ; Yang Deficiency ; chemically induced ; immunology

UNLABELLEDTo investigate the regulatory effects of epimedium flavonoids (EF) on adrenocortical regeneration in rats with inhibited hypothalamic-pituitary-adrenal (HPA) axis.

METHODSCell distribution in cell cycle and cell apoptotic rate were measured with PI stain and flow-cytometry; apoptosis cells were showed by in situ terminal-deoxynucleotidyl transferase-mediated deoxy-uridine triphosphate-fluorescene nick end labeling assay (TUNEL), and the genome-wide gene mRNA expression was detected by oligonucleotide microarrays.

RESULTSCompared to the normal control, adrenal cells isolated from the HPA axis inhibited model group were arrested in Go/GI phase, and showed a higher apoptotic rate (P < 0.05). After treated with EF, cells in G0/G1 phase decreased and those in G2/M phase increased (P < 0.01), and the elevated apoptotic rate reduced significantly (P < 0.05). TUNEL assay showed the number of apoptotic cells per section was 4.67 1.53 in the normal control group, 70.67 +/- 9.29 in the model group, and 18.67 +/- 7.64 in the EF-treated group respectively (n=3). Gene expressions in adrenal were mostly restrained in the model group, including 7 cytocycle promoting genes, including V-ras, V-jun, etc., while after treatment with EF, 6 cytocycle promoting genes, 1 anti-apoptotic gene, and genes that closely related with adrenocortical regeneration as IGF-II and FGF7 and their receptors, as well as 7 steroid biosynthesis participated genes were all up-regulated. Conclusion EF can accelerate adrenocortical cell proliferation, inhibit its apoptosis, and promote steroid biosynthesis so as to enhance adrenocortical regeneration in HPA axis inhibited rats, which may contribute to the beneficial effects of EF in protecting adrenocortical function during glucocorticoid withdrawal.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression ; Gene Expression Profiling ; Glucocorticoids ; adverse effects ; Hypothalamo-Hypophyseal System ; drug effects ; Male ; Oligonucleotide Array Sequence Analysis ; Pituitary-Adrenal System ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; prevention & control

OBJECTIVETo study the regulatory pathways and rules of the gene networks in Shen deficiency syndrome.

METHODSTissues of hypothalamus, pituitary, adrenal, lymphocyte, bone, liver and kidney were taken as samples from 4 months' and 24 months' old SD rats and rats after treatment with Epimedium flavonoids (EF), differences of gene expression profile in Shen deficiency syndrome were studied repeatedly with gene chip rat expression set U230 2.0 array from Affymetrix Co.

RESULTSGene expressions in the aged rats all decreased including neurotransmitter of gamma-aminobutyric acid (gammaGABA), gonadotropin releasing hormone (GnRH), thyrotropin-releasing hormone (TRH), thyroid stimulating hormone (TSH), growth hormone-releasing hormone receptor (GHRH), insulin-like growth factor (IGF) and binding proteins (IGFBP) in hypothalamus, pituitary and adrenal (HPA axis), cell growth-related gene, growth factor related protein, and immune regulatory genes such as interferon gamma (IFN-gamma), interleukin 4 (IL-4) and interleukin 6 (IL-6) in lymphocytes, parathyroid hormone (PTH), calcitonin, procollagen, collagen, connective tissue growth factor in bone, and oxidative phosphorylation genes such as cytochrome P450 and NADH dehydrogenase, glutamate dehydrogenase related with protein metabolism, and glucose-6-phosphatase related with glucose metabolism in liver, most of which were up-regulated after treatment with EF as well as genes related with ageing and cell cycle, such as cyclin B, metabolism related genes and proteins of sodium and chloride channel in kidney.

CONCLUSIONDysfunction of the two regulatory pathways of gene networks as nerve-endocrine-immunity and nerve-endocrine-bone metabolism exists in Shen deficiency syndrome differentiated by effects of drugs, which could be improved by strengthening Shen therapy.

Aging ; genetics ; physiology ; Animals ; Diagnosis, Differential ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression ; Hypothalamo-Hypophyseal System ; drug effects ; physiology ; Kidney Diseases ; genetics ; physiopathology ; Male ; Medicine, Chinese Traditional ; Neurosecretory Systems ; drug effects ; physiology ; Pituitary-Adrenal System ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; genetics ; physiopathology

OBJECTIVETo investigate the regulation pattern of the two compound prescriptions for Kidney tonifying, Yougui Yin and Bushen Yishou capsule, in down-regulating T-cell apoptosis gene expression in aged rats.

METHODSExpressions of T-cell apoptosis promoting and inhibiting genes, including Fas, FasL, Bcl-2, Bax, TNFR1 and TNFR2, as well as activity of cysteine proteinase in cascade connection, such as Caspase 8 and Caspase 3 were determined by TUNEL labeled flow cytometry and fluorescence real-time quantitative RT-PCR technique. The difference between old and young rats was compared, and the different regulation patterns of the two compound prescriptions for Kidney tonifying and their effects on Caspase activity were compared with those of compound prescription for blood circulation activating.

RESULTSThe two compound prescriptions for Kidney tonifying could effectively lower T-cell over-apoptosis in old rats, down-regulate FasL and TNFR1 gene transcription, and decrease the activity of Caspase 8 and Caspase 3, while the compound prescription for blood circulation activating showed insignificant effect on T-cell over-apoptosis.

CONCLUSIONKidney-deficiency is closely related to the T-cell over-apoptosis. The T-cell over-apoptosis in old rats could be effectively improved by the two compound prescription for Kidney tonifying through down-regulating the apoptosis promoting genes FasL and TN-FR1 transcription. That is the unique regulation pattern of Kidney tonifying principle to T-cell apoptosis related gene in old rats.

Aging ; drug effects ; genetics ; immunology ; Animals ; Apoptosis ; drug effects ; Blood Circulation ; physiology ; Caspase 3 ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; Female ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Random Allocation ; Rats ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; T-Lymphocytes ; cytology ; Yang Deficiency ; immunology ; fas Receptor ; biosynthesis ; genetics

Objective To analyze the epidemiologic characteristics of epidemic hemorrhagic fever syndrome (EHF) from 2004 to 2010 and provide scientific basis for formulating control measures.Methods Epidemiological data of EHF between 2004 to 2010 were obtained from the“National Disease Surveillance Report on Management Information System”,and the population data were from the“National Disease Surveillance Information Management System for Basic Information Report System”.Descriptive epidemiological methods was used for statistical analysis.ResultsA total of 173 cases were reported in Qian'an from 2004 to 2010.From 2004 to 2010,the cases were 86,67,12,1,0,1,and 7 cases,respectively,and the rate were 13.12/10 million ( 86/640 249 ),10.42/10 million (67/642 688 ),1.86/10 million ( 12/645 124),0.15/10 million (1/647 983 ),0(0/650 720),0.15/10 million( 1/653 839),and 0.11/10 million(7/657 380),respectively.The overall rate was 3.86/10 million(25/648 283) of population.From 2004 to 2008 the incidence reported declined rapidly,then increased slowly after 2009.The cases were found intensively in winter(November - next January) and spring season (february - may).The incidence in the age group of 10 - 45 was higher than that of other age groups,and the total number of cases was 82.08％(142/173).The incidence in males( 114 cases) was higher than that of females(59 cases).Occupational distribution mainly to peasants and students,which accounted for 87.86％ (152/173).Conclusions Epidemic in the city declines rapidly follows by a slow recovery,suggesting that in the future surveillance,mice-killing and protection of vulnerable population should be strengthened.

OBJECTIVETo investigate the effects of tributyrin (TB), a histone deacetylase inhibitor, on the growth, differentiation and apoptosis of SHI-1 leukemia cells and explore its possible mechanism.

METHODCell proliferation and viability were determined by cell counting, trypan blue dye exclusion. Cell morphological analysis, Annexin binding, DNA electrophoresis, expression of CD11b and CD14, NBT reduction were performed to evaluate differentiation and apoptosis of SHI-1 cells. The level of acetylated histone H3 was detected by Western blot and p21(WAF1/CIP1) expression by semi-quantitative RT-PCR.

RESULTSTB inhibited the proliferation and viability of SHI-1 cells in a time-dose dependent manner. The morphology of SHI-1 cells cultured in the presence of 0.1 mmol/L TB for 72 hs was more mature with higher NBT positivity and up-regulated expressions of CD11b and CD14 than that of control group. Exposed to 0.5 - 1.0 mmol/L TB for 48 hs, SHI-1 cells exhibited the morphological hallmarks of apoptosis, the increasing of Annexin binding and the DNA ladder on gel electrophoresis. The level of acetylated histone H3 and p21(WAF1) mRNA extracted from SHI-1 cells were increased by the treatment of TB.

CONCLUSIONTB can inhibit proliferation, induce differentiation and apoptosis of SHI-1 cells. The mechanism may associate with its up-regulation of acetylated histone and the expression of p21(WAF1).

Acetylation ; drug effects ; Apoptosis ; drug effects ; Blotting, Western ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Enzyme Inhibitors ; pharmacology ; Gene Expression ; drug effects ; Histone Deacetylase Inhibitors ; Histone Deacetylases ; metabolism ; Histones ; metabolism ; Humans ; Leukemia, Monocytic, Acute ; genetics ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Triglycerides ; pharmacology

OBJECTIVETo evaluate the correlation between programmed death-ligand 1 (PD-L1) expression in primary lung cancer cells, tumor associated macrophages (TAM) and patients' clinicopathological characteristics.

METHODSFrom 2008 to 2010, 208 non-small cell lung cancer patients who underwent surgery or CT-guided biopsy were recruited from Huadong Hospital, Fudan University. Immunohistochemistry staining was performed to evaluate the PD-L1 expression in both primary lung cancer cells and CD68 positive TAM. The relationship between PD-L1 expression and the clinical pathology was evaluated using χ(2) test. Spearman's rank correlations were used to determine the correlation between PD-L1 expression in tumor cells and macrophages.

RESULTSPositive PD-L1 expression in primary cancer cells was found in 136 (65.3%) patients, which were negatively correlated with lymph node metastasis (P=0.009) and smoking history (P=0.036). Besides, TAM with PD-L1 expression (found in 116 patients) was positively associated with smoking history (P=0.034), well-differentiation (P=0.029) and negative lymph node metastasis (P=0.0096). A correlation between PD-L1 expression in primary tumor cells and non-small cell lung cancer associated macrophages was found (r=0.228, P=0.021).

CONCLUSIONPD-L1, secreted from TAM, might induce cancer cells apoptosis, and decrease lymph node metastasis.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; B7-H1 Antigen ; secretion ; Carcinoma, Non-Small-Cell Lung ; pathology ; secretion ; Cell Line, Tumor ; Female ; Humans ; Lung Neoplasms ; pathology ; secretion ; Lymphatic Metastasis ; Macrophages ; pathology ; secretion ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo explore similarity and difference of connotation between Shen deficiency syndrome (SDS) and Shen-yang deficiency syndrome (SYDS) on the molecular level.

METHODSThe senescent SD rats and corticosterone-treated rats were adopted for models of SDS and SYDS respectively, their syndrome was differentiated according to the therapeutic efficacy of treatment with epimedium flavonoids (EF). The gene expression profiles of hypothalamas, pituitary, adrenal gland and lymphocytes (HPAT axis) were detected before and after EF treatment using gene chip provided by Affymetrix company.

RESULTSAs compared with the young rats, the ageing rats and corticosterone-treated rats showed a significant down-regulation in highly consistent pattern, of various neurotransmitters of HPAT axis firstly, followed with that of growth and sex hormone related genes. EF could reverse the above genes expression in both models, and for SYDS model rats, it could also significantly up-regulate the gene expressions of heat shock protein, cytochrome P450 and thyroid stimulating hormone (TSH).

CONCLUSIONBoth SDS and SYDS model rats show connotation of Shen deficiency, and the substantial base of Shen-yang deficiency syndrome resides in the process of oxidative phosphorylation of energy metabolism accelerated by thyroid hormone.

Adrenal Glands ; drug effects ; metabolism ; Animals ; Diagnosis, Differential ; Epimedium ; chemistry ; Flavonoids ; therapeutic use ; Gene Expression Profiling ; Hypothalamus ; drug effects ; metabolism ; Lymphocytes ; drug effects ; metabolism ; Male ; Medicine, Chinese Traditional ; Oligonucleotide Array Sequence Analysis ; methods ; Pituitary Gland ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Time Factors ; Yang Deficiency ; diagnosis ; drug therapy ; genetics