The morphological index of the seedlings including the plants height, the ground diameter, the leaf amounts, the fresh weight of the whole plant and the ratio of height to diameter was measured and the principal components were analyzed so as to determine the grading index, and stepwise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, the yield and the quality. The results showed that the height and ground diameter were determined as the quality indicators of the seedlings grading and the standard quality grading of seedlings of Anoectochilus roxburghii was initially set up, different seeding plants influenced the plants growth and the yield. The ground diameter of the class I was larger than that of the class II and III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal materials. The results of the study provide the basis for standard cultivation of A. roxburghii.
Drugs, Chinese Herbal ; analysis ; Orchidaceae ; chemistry ; classification ; growth & development ; Quality Control ; Seedlings ; chemistry ; classification ; growth & development

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.
Breeding ; methods ; Culture Media ; chemistry ; pharmacology ; Orchidaceae ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Survival Analysis

Mycena subpiligera, a new taxon in sect. Fragilipedes that can strongly enhance the germination efficiency of Gastrodia elata seeds, was discovered in subtropical areas of China. As revealed by a morphological comparison with related Mycena species as well as maximum likelihood (ML) and Bayesian phylogenetic analyses based on sequences of the internal transcribed spacer (ITS) and the large subunit (LSU) regions of nuclear ribosomal RNA, the new taxon can be distinguished from phenotypically similar and phylogenetically related species. Optimal cultural conditions for M. subpiligera basidiomata are reported, and the germination rate of the new species is compared with that of M. citrinomarginata.

OBJECTIVETo investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs.

METHODS① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit.

RESULTS①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002].

CONCLUSION①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.

Bone Marrow Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; immunology ; Humans ; Immune Tolerance ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; immunology ; T-Lymphocytes ; cytology

Whole-cell patch clamp recording was used to investigate the action of beta-amyloid peptide(1-40) (Abeta(1-40)) on high voltage-activated calcium channel current (I(HVA)) in acutely isolated hippocampal CA1 pyramidal neurons in rats and observe its modulation by ginkgolide B (GB). Drug was applied by extracellular bath or adding in the pipette solution, and its effect was determined by comparing the amplitude of I(HVA) before and after the drug application. Bath application of aggregated Abeta(1-40) at concentrations of 0.01~30 mumol/L increased the amplitude of I(HVA) in a dose-dependent manner by (5.43+/-3.01)% (n=8, P>0.05), (10.49+/-4.13) % (n=11, P>0.05), (40.69+/-8.01) % (n=16, P<0.01), (58.32+/-4.85) % (n=12, P<0.01), and (75.45+/-5.81) % (n=6, P<0.01), respectively, but had no effect on the I-V curve of I(HVA); fresh Abeta(1-40) almost had no effect on I(HVA) (n=5, P>0.05). L-type calcium channel antagonist nifedipine abolished the increase of I(HVA)by Abeta(1-40). The increase of I(HVA) by Abeta(1-40) (1.0 mumol/L) was enhanced to (66.19+/-5.74) % (P<0.05) by 8-Br-cAMP (membrane permeable analogue of cAMP) and to (73.21+/-6.90) % (P<0.05) by forskolin, an adenylyl cyclase (AC) agonist, and reduced to (20.08+/-2.18) % (P<0.05) by H-89, cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) antagonist. GB effectively inhibited the increase of I(HVA) by Abeta(1-40). The results indicate that Abeta(1-40) leads to an intracellular calcium overload by increasing I(HVA) via AC-cAMP-PKA. This may be one of the mechanisms for its neurotoxicity. GB can prevent neurons from neurotoxicity by inhibiting abnormal calcium influx caused by Abeta(1-40).
Amyloid beta-Peptides ; toxicity ; Animals ; Animals, Newborn ; Calcium Channels ; drug effects ; Ginkgolides ; pharmacology ; Hippocampus ; cytology ; metabolism ; Lactones ; pharmacology ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; Patch-Clamp Techniques ; Peptide Fragments ; toxicity ; Rats ; Rats, Wistar

OBJECTIVETo explore the mechanisms of differentiation and development of pancreatic endocrine cells as well as pancreatic regeneration.

METHODSHuman embryonic pancreatic tissue at 7-14 weeks of gestation was collected. Diabetes mellitus rat model was induced with 65 mg/kg of streptozotocin. Insulin, glucagon, somatostatin, nestin, and cytokeratin 19 (CK19) of pancreatic tissues were observed by immunohistochemistry.

RESULTSAt 9 weeks of gestation, pancreatic epithelial cells began to co-express insulin, glucagon, somatostatin, and CK19 before migration. Islet cells gradually congregated along with the increase of aging, and at 14 weeks of gestation histological examination showed islet formation. At 12 weeks of gestation, nestin-positive cells could be seen in the pancreatic mesenchyme. During early embryogenesis, islet cells of pancreatic ducts co-expressed insulin, glucagon, and somatostatin. During pancreatic regeneration after damage, nestin expression of islet cells increased.

CONCLUSIONIn the early stage of embryogenesis, islet cells of primary pancreatic ducts can be differentiated to multipotential endocrine cells before migration. During tissue regeneration, pancreatic stem cells may differentiate and proliferate to form pancreatic islet.

Animals ; Cell Differentiation ; Diabetes Mellitus, Experimental ; chemically induced ; metabolism ; pathology ; Embryonic Development ; physiology ; Epithelial Cells ; cytology ; physiology ; Humans ; Insulin-Secreting Cells ; cytology ; physiology ; Islets of Langerhans ; cytology ; physiology ; Male ; Pancreas ; cytology ; embryology ; physiology ; Pancreatic Ducts ; cytology ; embryology ; physiology ; Rats ; Rats, Sprague-Dawley ; Regeneration ; physiology ; Stem Cells ; cytology ; metabolism ; physiology

To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.
Animals ; Brain Ischemia ; drug therapy ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate the expression of JWA after hemin and (or) thermal stress exposure.

METHODSThe expression of JWA was determined by Western blot. RT-PCR was carried out for evaluation of the expression of JWA mRNA. The JWA promoter transcription activity analysis was performed by CAT-ELISA.

RESULTSThe expression of JWA protein was significantly increased by 3.23 +/- 0.57 times of control in K562 cells after treated by hemin (50 micromol/L) for 1 week, and the similar pattern was observed in the cells after treatment with thermal stress (42 degrees C) for 2 hours (increased by 8.00 +/- 1.73 times). The expression of JWA mRNA was also significantly elevated by 1.37 +/- 0.06 times in K562 cells treated by hemin (30 micromol/L) for 48 hours, and with a similar regulation pattern (increased by 1.87 +/- 0.13 times) by treatment with thermal stress (42 degrees C) for 30 minutes. However, an antagonistic effect was observed by treatment of K562 cells with hemin (30 micromol/L, 48 hours) followed by thermal stress (42 degrees C, 30 minutes). The CAT-ELISA further confirmed that hemin or thermal stress treatment alone up-regulated JWA transcription activity while the effects could be counteracted partly by the combined treatment of the both.

CONCLUSIONThe hemin and thermal stress may regulate JWA expression via distinct intracellular signal transduction pathways.

Blotting, Western ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Heat Stress Disorders ; metabolism ; Heat-Shock Proteins ; genetics ; metabolism ; Hemin ; pharmacology ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo provide guidance for selection and breeding of Pesudostellaria heterophylla, agronomic traits of 3 mainly cultivated form of P. heterophylla were observed and compared in Guizhou shibing.

METHODThirteen agronomic traits of P. heterophylla from 3 cultivation form were measured and the traits were analyzed using multiple comparison, correlation analysis, multiple stepwise regression analysis and path analysis.

RESULTThe results showed that agronomic traits were different in 3 cultivation form, significant(P <0.05)or very significant correlated (P <0.01)among multiple agronomic traits, the width and the number of root, the first branch number, cleistogamous flowers, length and width of leaves, and the aboveground biomass were the main factors that affected the underground biomass, and the number of root, the aboveground biomass had a directly positive effect on the underground biomass. Meanwhile, whole length, length and width of leaves, cleistogamous flowers etc. had direct or indirect effect on the underground biomass.

CONCLUSIONAboveground biomass would be as the best indirect selection traits on breeding high yield of P. heterophylla, the first branch number, width of leaves and cleistogamous flowers world be as a better auxiliary index on breeding high variety of P. heterophylla.

Agriculture ; economics ; Animals ; China ; Hemiptera ; anatomy & histology ; growth & development ; Regression Analysis

AIMTo study the formation and localization of ADM mRNA in lung tissues and investigate the effects of ADM on isolated tracheal strip contraction induced by histamine in asthmatic guinea pig.

METHODSThe guinea pigs (n = 22) were randomly divided into two groups of 11 each: asthmatic group and control group. The formation and localization of ADM mRMA were observed by in site hybridization. The effect of exogenous ADM on contractions of isolated tracheal strip of the asthmatic guinea pigs to histamine was examined.

RESULTSThere were strong positive expression for ADM mRNA in airway epithelial cells (AEC), smooth muscle cells (ASMC) in asthmatic group. The control group showed significantly decreased number of ADM mRNA positive cells in lung tissues. From 10(-11) mol/L to 10(-7) mol/L, ADM may cause concentration depend pentiation of the isolated tracheal strip contraction induced by histamine of asthmatic group which was higher significantly compared the control group (P < 0.05). 10(-8) mol/L ADM reached the maximal relaxation, with the increasing of ADM, neither asthmatic nor control group can increase the relaxation.

CONCLUSIONThere is ADM mRNA overproduction in AEC and ASMC and exogenous ADM may inhibit isolated tracheal strip contraction induced by histamine of asthmatic guinea pig, which may contribute to airway inflammation and hyperresponsiveness in asthma.

Adrenomedullin ; metabolism ; Airway Resistance ; Animals ; Asthma ; metabolism ; physiopathology ; Guinea Pigs ; In Vitro Techniques ; Lung ; metabolism ; Male ; Trachea ; physiopathology