In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

Aim To investgate the mechnism through which ginkgolides affect learning and memory capabilities of the Alzheimers disease-like rats. Methods Okadaic acid(OA)was injected into the CA1 region of the rat hippocampus and the rats were gavaged with ginkgolides. The learning and memory abilities of the rats were assessed through Morris water maze behavioral test, and the expressions of nicotinic acetylcholine receptors and ChAT were observed by Western blotting and immunohistochemistry, respectively.Results Compared with the control rats, the capabilities of learning and memory were lowered significantly(P

Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual funetion.However,the color vision system of animals which encounter uncertainty of spectral signals should be plastic.Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear.Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light.Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed tO the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively.The flat-mounted retinal sample was prepared and divided into dorsal zone,ventral zone and mixed zone anatomically according to the distribution of difierent light-sensitive cone.The changes in density of cone cells sensitivited to different colored light were detected by single-1abel or double-label immunocytochemistry.The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively.Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group,purple light group and white light group (F=234.28,P<0.01).Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group wag evidently lower(q=389.68,P<0.01；q=67.11,P<0.01).No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group,purple light group and white light group(F=3.14,P>0.05).The density of coexpression of the mixed cone cells was increased in green light group(q=157.55,P<0.01)but decreased in purple light roup (q=254.85,P<0.01)in comparison with white light group.The expression levels of green-opsin and green-opsin mRNA in green light group was significantly elevated(q=184.45,P<0.01；q=4.71,P<0.05),but those of purple light group were evidently declined(q=5.87,P<0.05；q=346.66,P<0.01)in comparison with white light group.There was no statistically significant differences were found in the expression of purple-opsin and its mRNA among all the groups(F=1.24,P>0.05；F=3.27,P>0.05).Conclusion After the exposure of long-time monochromatic light illumination,monochromatic cones density and its opsin in guinea pig occur the corresponding alteration to gain good spatial vision as a compensatory reaction.These outcomes imply that there is some plasticity during the development of color vision.The increase of green-sensitive cones might be from the differentiation of coexpression cones in transition region.

Bronchiolitis ; diagnosis ; microbiology ; pathology ; Female ; Humans ; Infant ; Leukotriene D4 ; analysis ; Male ; Nasopharynx ; chemistry ; microbiology ; pathology

Objective To investigate the relationship between myocardial ischemia and slow coronary flow phenomenon with 99Tcm-methoxyisobutylisonitrile (MIBI) adenosine myocardial perfusion SPECT imaging. Methods Forty-four patients were divided to three groups according to the result of coronary angiography(CAG). There were GAG-positive(P-GAG) (n=12),slow coronary flow (CSF) (n =22),and normal coronary flow (NCF) (n = 10). Results of adenosine myocardial perfusion imaging were compared among these three groups. Semi-quantitative visual scoring method was used to evaluate the myocardial perfusion:0 = normal,1 = mild decrease,2 = moderate decrease,3 = severe decrease,4 = defect. Statistical analysis was performed using variance analysis,t-test and x2-test. Results No significance was observed at age ( t =0.27,0. 54 and 0. 59),sex (x2 = 0. 92),hypertension,hyperlipemia and diabetes (x2 = 1.23,all P ＞ 0.05 ) among the three groups. A significantly higher frames of the coronary thrombolysis in myocardial infarction (TIMI) flow was noted in CSF than in NCF groups (33.7 ±5.5 vs 17.6 ±3.9,t = 9. 58,P ＜0. 001 ). The positive adenosine myocardial perfusion imaging rate were significant among these three groups with 100% (12/12) in P-CAG group,77.3% (17/22) in CSF group,and 20% (2/10) in NCF group. When using semi-quantitative visual scoring method,significantly higher average ischemia segments were noted in CSF group than in NCF group ( 1.06 ± 0.77 and 0. 91 ± 0.80,t = - 2. 02,P ＜ 0. 05 ),but was less than that in P-CAG group (2.41 ±0.79,t =4. 54,P ＜0.001 ). The degree of ischemia of CSF group was higher than that in NCF group ( 8.01 ± 6.06,and 2.73 ± 2.60,t = - 2.07,P ＜ 0.05 ) and was less than that in P-CAG group (14. 07 ±12. 77 ,t=1.44,P＞0. 05). Conclusion Slow coronary flow phenomenon can be detected by adenosine myocardial perfusion image to offer the evidence of diagnosis and treatment for the chest pain patients with negative coronary angiography results.

P13-OMP (29.1). P13-OMP and OMP68 group challenged with P13 and P11 can be efectivly protected; P13-WCB group challenged with P13 and P11 can not be efectivly protected; the control group were died out. The P13-OMP and OMP68 of Bordetella bronchiseptica has good immunogenicity and protection, so the results of this study lay good theoretical foundation for OMP subunit vaccine.

Programmed death-1 ligand-1(PD-L1) is a recently identified member of the B7 family molecules and is shown to mediate the inhibition of immune responses. This study was purposed to enhance the weak immunological function of dendritic cells (DCs) derived from the patients with chronic myelocytic leukemia (CML) by blockade of the expression of PD-L1. Bone marrow mononuclear cells (BMMNCs) of CML patients were induced into DCs in the presence of cytokine cocktail of rhGM-CSF, rhIL-4 and TNF-alpha. The phenotypes of DCs were detected by flow cytometry, mixed lymphocyte reaction was analyzed by MTT assay and IFN-gamma, IL-2 and IL-10 in the cell culture supernatant were detected by ELISA. The results showed that the expression of PD-L1 on CML-DCs was upregulated with the maturation of CML-DCs. PD-L1-blockaded DCs could enhance T lymphocyte proliferation, increase the secretion of IL-2 and IFN-gamma, and inhibit the production of IL-10. Taken together, PD-L1-blockaded DCs originated from CML cells had more potent immunostimulatory capability. It is concluded that PD-L1 blockaded can enhance the function of CML-DCs. This approach presents new possibilities for achieving anti-tumor immunity by DC-based vaccination.
Antigens, CD ; metabolism ; B7-H1 Antigen ; Dendritic Cells ; cytology ; immunology ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; metabolism ; Recombinant Proteins ; Tumor Necrosis Factor-alpha ; pharmacology

A quantitative analytical method of ultra-high performance liquid chromatography (UPLC) was developed for simultaneously determining twelve components in Tibetan medicine Zuozhu Daxi. SIMPCA 12.0 software was used a principal component analysis PCA) and partial small squares analysis (PLSD-DA) on the twelve components in 10 batches from four pharmaceutical factories. Acquity UPLC BEH C15 column (2.1 mm x 100 mm, 1.7 µm) was adopted at the column temperature of 35 °C and eluted with acetonitrile (A) -0.05% phosphate acid solution (B) as the mobile phase with a flow rate of 0. 3 mL · min(-1). The injection volume was 1 µL. The detection wavelengths were set at 210 nm for alantolactone, isoalantolactone and oleanolic; 260 nm for trychnine and brucine; 288 nm for protopine; 306 nm for protopine, resveratrol and piperine; 370 nm for quercetin and isorhamnetin. The results showed a good separation among index components, with a good linearity relationship (R2 = 0.999 6) within the selected concentration range. The average sample recovery rates ranged between 99.44%-101.8%, with RSD between 0.37%-1.7%, indicating the method is rapid and accurate with a good repeatability and stability. The PCA and PLSD-DA analysis on the sample determination results revealed a great difference among samples from different pharmaceutical factories. The twelve components included in this study contributed significantly to the quantitative determination of intrinsic quality of Zuozhu Daxi. The UPLC established for to the quantitative determination of the twelve components can provide scientific basis for the comprehensive quality evaluation of Zuozhu Daxi.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Tibet

OBJECTIVETo study the protective effects of salidroside on injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cell.

METHODApoptosis and intracellular free calcium concentration ([Ca2+]i) were measured by flow cytometry, morphological changes and neuronal necrosis were observed with fluorescence microscope, and the lactic dehydrogenate (LDH) release was measured by LDH kits.

RESULTHypoxia/hypoglycemia cultures for 2 hours induced neuronal apoptosis and necrosis. They were 18.59% (P < 0.01) and 4.94% (P < 0.01) respectively. There were morphological changes including chromatin condensation, nuclear fragmentation and formed apoptotic bodies after exposed to hypoxia/hypoglycemia for 2, 4, 6, 12 hours. After 2 hours of hypoxia/hypoglycemia, neuronal [Ca2+]i and the release of LDH were significantly increased. They were 8.46 nmol/L (P < 0.01) and 16.59% (P < 0.01) respectively. The effects were enhanced with the extending time of hypoxia/hypoglycemia. Salidroside might have significantly decreased the percentage of neuronal apoptosis and necrosis, reduced neuronal [Ca2+]i and the release of LDH. The effects of salidroside were strengthened with the increasing of Salidroside dosage.

CONCLUSIONSalidroside has effect of anti-neuronal apoptosis. This effect might be related to its function of decreasing intracellular free calcium concentration.

Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Hypoglycemia ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Phenols ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhodiola ; chemistry

To investigate the activity of CKLF1 on the proliferation and differentiation of bone marrow cells. Methods Human low density bone marrow cells and mouse bone marrow cells were plated in 96-well microplate and supernatants from transfected COS-7 cell culture were added. The cell proliferation was assayed by MTT method after 5 days incubation. The enhancing effect of CKLF1 on the colony forma tion of human hematopoietic progenitor cells was identified in semi-solid culture. Results CKLF1 has obvi ous enhancing effect on both human and mouse bone marrow cells, it can stimulate the colony formation of human hematopoietic stem cells and has synergistic action with GM-CSF. Conclusion CKLF1 can pro mote the proliferation and differentiation of bone marrow cells.