2.Study on the Optimum Culture for Growth of Bacillus Subtilis B115 andthe Antibacterial Effect of B115 on Aeromonas
Zhi-Hua SHEN ; Jin-Yu SHEN ; Wen-Lin YIN ; Xiao-Yi PAN ; Ying-Lei WU ;
Microbiology 1992;0(04):-
The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.
3.Effect of medicines for activating blood and reinforcing Qi on angiogenesis in infarcted myocardium edge area of acute myocardial infarction model in rats.
Wen-Hua ZANG ; Shen-Hua YIN ; De-Cai TANG ; Bing-Bing LI
China Journal of Chinese Materia Medica 2014;39(5):901-906
OBJECTIVETo study the effect of medicines for activating blood and reinforcing Qi on the number of new micro-vessels and the protein expressions of VEGF and bFGF in the infarcted myocardium edge area of acute myocardial infarction (AMI) model in rats.
METHODThe AMI model of rats was established. After the successful model establishment, rats were randomly divided into the sham-operated group, the model group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group, the Chuanxiong-Huangqi (1 : 2) group, the Danshen group, the Chuanxiong group, the Chishao group and the Shexiang Baoxin pill group, with five rats in each group. Rats in each medicated group were orally administered with drugs as per 13.5 g x kg(-1) x d(-1) once everyday for three weeks. The immunohistochemical SP method was adopted to detect the expression of vWF in myocardial tissues, and count the number of micro-vessels (MVC). The protein expression of VEGF and bFGF in myocardial tissues were determined by Western blot.
RESULTThe new micro-vessels stained by vWF factor could be found in the infarcted myocardium edge area of the sham-operated group, the model group and all of medicated groups. The sham-operated group show unobvious new micro-vessels in myocardial tissues. A small amount of new micro-vessels could be seen in the infarcted myocardium edge area of the model group. Whereas a larger number of micro-vessels could be seen in the infarcted myocardium edge area of all of medicated groups. The differences between the sham-operated group and the model group had statistical significance (P < 0.05). The differences between each medicated group and the model group had statistical significance as well (P < 0.05 or P < 0.01). The lowest protein expression of VEGF and bFGF was found in myocardium of the sham-operated group, with the statistical significance compared with the model group (P < 0.05). Compared with the model group, each medicated group showed significant increase in the protein expression of VEGF and bFGF, with the statistical significance between them (P < 0.05 or P < 0.01).
CONCLUSIONThe Danshen group, the Chuanxiong group, the Chishao group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group and the Chuanxiong-Huangqi (1 : 2) group show the effect in promoting angiogenesis. Their mechanism for promoting angiogenesis may be related to the improvement of the protein expressions of VEGF and bFGF, so as to increase the contents of VEGF and bFGF and promote the angiogenesis of new vessels.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Humans ; Male ; Microcirculation ; drug effects ; Microvessels ; drug effects ; physiopathology ; Myocardial Infarction ; drug therapy ; physiopathology ; Neovascularization, Pathologic ; drug therapy ; genetics ; metabolism ; Qi ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism
4.Investigation of influence of 16-slice spiral CT electrocardiogram-controlled dose modulation on exposure dosage and image quality of cardiac CT imaging under simulated fluctuant heart rate
Yan YIN ; Jie CHEN ; Weiming CHAI ; Jia HUA ; Yun SHEN ; Na GAO ; Jiantong XU
Chinese Journal of Radiology 2008;42(10):1035-1039
Objective To investigate the influence of electrocardiogram(ECG)-controlled dose modulation on exposure dosage and image quality of cardiac CT imaging in a cardiac phantom with simulated fluctuant heart rate.Methods The basal heart rate of the cardiac pulsating phantom was set as 60 bpm.the experimental situations were divided as 6 groups according to different heart rates.The cardiac imaging was performed on the cardiac phantom when the ECG-controlled dose modulation was firstly turned off.The exposure dosage of each scan sequence was documented.The standard deviation of the CT values of the phantom was measured on the central slice after coronal reformation of the raw data.The quality of 2D and 3D images were scored.Thell cardiac imaging was performed when ECG modulation was on and set as four groups according to different modulation parameters.All the data were documented as before.The results from the five groups with and without ECG modulation current were analyzed bv F test and comparative rank sum test using the statistical software SPSS 10.0.Results Statistical analysis showed no significant difference(P>0.05)between the SNR of images(SD value was 27.78 and 26.30)from the groups that full mA output at wide reconstruction phase(69%~99%)when the heart rate was fluctuant(≥7.5 bpm).There was also no significant difference(P>0.05)between the quality of the 2D and 3D images.But there was a significant difference(P<0.01)between the SNR of images(SD value was 26.78 and 29.90)that full mA was only used at 85%reconstruction phase when the heart rate Was fluctuant(≥7.5 bpm).The exposure dosage was remarkably reduced when the ECG modulated current was on than when it Was off under fluctuant heart rate.Furthermore.there were significant difierence(P<0.01)among the difierent ECG modulated current parameter groups.The exposure dosage can be reduced by 44.7%under the situation that the heart rate was fluctuant.Whell the fluctuation of the heart rate was≤12.5 bpm,there wag no obvious relationship between the fluctuation of the heart rate and the exposure dosage (the variation was from 0.1 to 1.1 mSv),but if the heart rate fluctuation was>12.5 bpm,the exposure dosage would increase obviously (from 0.6 to 1.7 mSv).Conclusion For cardiac imaging with 16-slice row CT,the application of ECG modulated current can effectively reduce the exposure dosage without compromising the image quality even if heart rate was fluctuant.
5.Study on application of mix recombinant antigen in schistosomiasis diagnosis
Xuren YIN ; Chuanxin YU ; Yongliang XU ; Linnan SHEN ; Wanquan HUA ; Jian LI
Chinese Journal of Schistosomiasis Control 1992;0(06):-
Objective To investigate the value of mix recombinant antigen in schistosomiasis diagnosis. Methods The recombinant antigens of SjC23 (HD),SjC21.7 and SjCMP10 were expressed in vitro and purified by the affinity chromatography method. The efficacies of soluble egg antigen (SEA),single recombinant antigen and mix recombinant antigen for schistosomiasis diagnosis by Enzyme-linked Immunosorbent Assay were compared. Results The diagnostic efficacy was the same when the antibody IgG of the same group sera of schistosomiasis was detected by different quantities of 2.5 ?g/ml and 7.5 ?g/ml of SEA immobilized on microplate, and their absorbency A was the same, but there was a significant difference in the diagnostic efficacies between single recombinant antigen and mix recombinant antigen when the antibody IgG of the same group sera of schistosomiasis was detected by the same quantity of single recombinant antigen or mix recombinant antigen immobilized on microplate, the absorbency A of mix antigen reacted with the sera of schistosomiasis was significant higher than that of the single recombinant. The positive rates were very similar when 39 sera of acute schistosomiasis,80 sera of chronic schistosomiasis and 27 sera of advanced schistosomiasis were detected by SEA or mix recombinant antigen by ELISA in the same time. No cross-reaction presented when 20 clonorchiasis sera were detected by the mix recombinant antigen and no false positive presented when 40 of healthy sera were detected by the mix recombinant antigen. Conclusion The schistosomiasis diagnostic method by using the mix recombinant antigen has been established, which is helpful for improving the efficacy of schistosomiasis diagnosis.
6.Exploration on molecular mechanism of Epimediun flavonoids in regulating adrenocortical regeneration in rats with inhibited hypothalamic-pituitary-adrenal axis using oligonucleotide microarrays.
Jian-hua HUANG ; Zi-yin SHEN ; Wei-hua CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(5):423-426
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
7.Syndrome differentiation through drug effects in mapping the two regulatory pathways of gene networks in Shen deficiency syndrome.
Zi-yin SHEN ; Yu CHEN ; Jian-hua HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(6):521-525
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
8.Molecular mechanism of epimedium flavonoids in immune homeostasis remodeling in aged rats revealed by lymphocyte gene expression profile.
Yu CHEN ; Zi-yin SHEN ; Wei-hua CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(1):59-62
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
9.Effect and mechanism of Epimedium flavanoids for aging retardation from viewpoint of transcriptomics and metabonomics.
Jian-Hua HUANG ; Zi-Yin SHEN ; Bin WU
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(1):47-50
OBJECTIVETo investigate the effect and its mechanism of Epimedium flavanoids (EF) in retarding aging with different systematic viewpoints.
METHODSHypothalamus, pituitary, adrenal and lymphocytes taken from 4-, 10-, 18-, 24-month old rats and from EF treated 24-month old rats were used to measure whole genome mRNA expression by gene array. Serum samples were used for metabonomic assay with high performance liquid chromatography. Using specific gene chip for NF-kappaB signaling pathway to detect the gene expression of the molecule related to that pathway in lymphocytes. Then, a neural network (NN) model was established upon the data obtained to quantitatively evaluate the degree of aging and the efficacy of drug intervention.
RESULTSGene expression of 199 genes showedan age-dependent pattern, most of which were reversed by EF, and the output of NN model showed that EF made the transcriptomics of 24-month old rats to 8-13 months. Seventeen metabolites among the 1,885 peaks detected were identified to have significant age-depending changes, and EF intervention reset the level of metabolites to a younger (18-month) level. The integral level of gene expression for NF-kappaB signaling pathway decreased significantly along with the increasing of age, and was significantly elevated by EF, NN model showed it approached to 10.5-month old.
CONCLUSIONPhenotype of aging at different levels demonstrates a common age-dependent trend; EF can reverse this age-dependent change at different levels in a synchronous manner.
Adrenal Glands ; drug effects ; growth & development ; metabolism ; Aging ; drug effects ; genetics ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; drug effects ; Hypothalamus ; drug effects ; growth & development ; metabolism ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; NF-kappa B ; blood ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; methods ; Pituitary Gland ; drug effects ; growth & development ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Time Factors ; Transcription, Genetic ; drug effects
10.Experimental study on effect of epimedium flavonoids in protecting telomere length of senescence cells HU.
Zuo-Wei HU ; Zi-Yin SHEN ; Jian-Hua HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(12):1094-1097
OBJECTIVETo investigate the mechanism of senescence delay of human diploid fibroblast (2BS) and protecting telomere length by epimedium flavonoids (EF).
METHODSThe drug sera of EF were used to treat the 2BS. The population doublings of 2BS cells were observed, the mRNA expression of p16 gene were determined by fluorescence real-time quantitative RT-PCR, the telomerase activation of 2BS cells were determined by TRAP-Hyb, the total retinoblastoma (Rb) and phosphorated Rb protein content were detected by ELISA, the telomere length of 2BS cells were determined by telomere restriction fragment (TRF) Southern blot assay.
RESULTSEF could significantly extend the population doublings of 2BS cells, the expression of p16 mRNA was decreased and the content of phosphorated Rb protein were increased by EF. The telomere lengthening of 2BS cells were improved by EF, but the telomerase was not activated.
CONCLUSIONIn senescence human fibroblasts 2BS cells, p16 gene mRNA expression increased, content of phosphorated Rb protein decreased and the telomere length of 2BS shortened, EF might delay the aging of cells through inhibiting the p16 gene expression, promoting the production of phosphorated Rb protein and to protect the length of telomere, but not activating the telomerase.
Animals ; Cells, Cultured ; Cellular Senescence ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Epimedium ; chemistry ; Fibroblasts ; cytology ; Flavonoids ; pharmacology ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Retinoblastoma Protein ; metabolism ; Telomerase ; biosynthesis ; Telomere ; genetics ; metabolism ; Transduction, Genetic