Objective To investigate the effect of manganese-enhanced MRI (MEMRI) at 7.0T for tracing nerve tracts in rat brain in vivo. Methods With brain stereotactic apparatus, 0.4 μl Mncl_2 with aqueous solution of 1 mol/L was injected into the right somatosensory cortex of 9 SD rats. MR scan was performed for tracing corticospinal tracts and other coherent nerve tracts pre-, and 24, 48, 72 h, 7 days post-injection with 7.0T micro-MRI system, respectively. Results Corticospinal tracts were showed in intact after Mn~(2+) administration from somatosensory cortex, thalamus, cerebral peduncle to pons at the time point of 24, 48, 72 h and 7 days, while the best tdisplaying was achieved at 24-48 h after Mn~(2+) administration. Simultaneously a small quantity of Mn~(2+) reached the opposite somatosensory cortex through the corpus callosum. Conclusion MEMRI for tracing rat nerve tracts can be showed clearly with 7.0T MRI. The location of manganese-enhanced corticospinal tracts in agreement with the rat brain atlas in stereotaxic is in agreement with that Paxinos' published. MEMRI can display the relationship between the two sides of hemisphere, and may play an important role in investigating the brain function and nerve plasticity after nerve injury in vivo.

Objective This study is undertaken to evaluate the changes of serum cytokine levels in different stages of collagen induced arthritis(CIA)rats,to search for the specific proteins related with rheuma- toid arthritis(RA)pathogenesis and inflammation,and to explore the mechanism of RA pathogenesis.Methods Rat cytokine antibody array coated with 19 specific cytokine antibodies was used to examine serum samples at peak and late stage of CIA rats,and were compared to normal cytokine levels.At the same time,ELISA assay for serum TNF-?production was used to verify the array results.Results Among the target cytokines,10 up- regulating cytokines were kept in high expression in different phases of disease,while 1 showed significant change only at the peak of disease.There was no downregnlating cytokines in the results.Serum TNF-?assay results were consistent to the array results.Conclusion Cytokines show different expression in CIA at differ- ent stages,and specific cytokines can be used as the candidates to further study of the RA pathogenesis.This study also provides molecular makers for early diagnosis.

AIM To observe the changes of plasma neuropeptide Y (NPY) levels in patients with essential hypertension(EH) taking Compound Qishao Jiangya Tablets (Radixet Rhizoma Notoginseng, Radix Paeoniae Alba, Rhizoma Gastrodiae, etc) and to investigate the significance of NPY in the pathogenesis of hypertension and the relationship between NPY and platelet disnormal function. METHODS The plasma NPY concentrations of 60 patients with EH and 20 normal ones were determined by radioimmunoassay, at the same time detected platelet aggregation rate and fibrinogen were detected. RESULTS The levels of plasma NPY in patients with EH were higher than that in normal control group. The NPY levels, the platelet aggregation rate, fibrinogen in the three periods of hypertension (period Ⅰ,Ⅱ, Ⅲ) were all significantly different and were increased with the severity of hypertension. CONCLUSION NPY may participate in the pathophysiological course of hypertension, and the plasma NPY, the platelet aggregation rate of the patients with EH were correlative. The plasma NPY should be dynamscally monitored in order to determine the disease degree. During the treatment for EH attention should be taken to decrease the platelet aggregation rate.

A new phenethanol, (2'R)-4-(2', 3'-dihydroxy-3'-methyl-butanoxy)-phenethanol (1), along with other eleven known benzene derivatives (2-12) were isolated from the roots, stems and leaves of Clausena excavata (Rutaceae). Compounds 3 and 4 are new natural products, and compounds 5-8, 10-12 were isolated from C. excavata for the first time. Their structures were elucidated on the basis of MS, 1D and 2D NMR spectroscopic analyses including HSQC, COSY and HMBC experiments. 1 was tested for its cytotoxicities against A549, HeLa and BGC-823 cancer cell lines, and antimicrobial activities against Candida albicans and Staphylococcus aureus. The results showed that 1 did not exhibit cytotoxic and antimicrobial activities.
Benzene Derivatives ; chemistry ; Candida albicans ; drug effects ; Cell Line, Tumor ; Clausena ; chemistry ; HeLa Cells ; Humans ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Staphylococcus aureus ; drug effects

To investigate monoterpenes and sesquiterpenes of the stems and leaves of Clausena excavata, an AcOEt fraction of the methanol extract was subjected on column chromatographies including silica gel and RP-18, as well as preparative HPLC. The structures of compounds isolated were identified on the basis of spectroscopic data as excamonoterpene (1), (6R, 9S)-9, 10-dihydroxy-4-megastigmen-3-one (2), (3R, 6R, 7E) -3-hydroxy-4, 7-megastigmadien-9-one (3), (3S) -3-hydroxy-7, 8-dihydro-beta-ionone (4), (3S, 5R, 6S) -3-hydroxy-5,6-epoxy-beta-ionone (5), (6R, 9R) -9-hydroxy-4-megastigmen-3-one (6), (3S, SR) -dihydroxy-6, 7-megstigmadien-9-one(7), (-)-loliolide(8), caryolane-1, 9alpha-diol(9) and 2, 6-dihydroxyhumula-3 (12), 7 (13), 9(E)-triene (10), were isolated from the stems and leaves of C. excavata. Compound 1 is a new monoterpene, named as excamonoterpene. Compounds 2-10 were isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; methods ; Clausena ; chemistry ; Magnetic Resonance Spectroscopy ; Methanol ; chemistry ; Molecular Structure ; Monoterpenes ; analysis ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Sesquiterpenes ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization

This work was designed to study a novel dry powder inhalation (DPI) carrier for drug loading and release of tiotropium bromide (asthma medicine). The synthesized lactose drug-carrier with a flower shape was crystalline. The carrier with a micro-meso-macroporous structure had advantages of high pore surface area, high capacity of drug loading and fast release of drug. In the study of loading tiotropium bromide, the drug was distributed at the core of carrier using the solution-based method, while the morphology was changed a little and the amount of loaded drug was 5% (w/w). Using the crystallization-based method, the drug was distributed at the shell of carrier, while the morphology was changed a lot and the amount of loaded drug was 49% (w/w). In addition, with the impact of carrier structure, the drug release rate was increased first and then decreased thereafter using the solution-based method, while the drug release rate was decreased first and then increased thereafter using the crystallization-based method. Thus, the lactose microparticles can be used as a novel drug carrier for dry powder inhalation.

Objective To investigate the possibility of using well-differentiated human airway epithelial cells (HAE) to isolate and identify human influenza A virus from a stale respiratory tract specimen.Methods The stale specimen used in this study was a nasopharyngeal swab specimen collected from a patient with unexplained pneumonia in Qinghai in 2010.It was positive for influenza A virus (H3N2) RNA, but negative for hemagglutination.Equal amount of the specimen was inoculated on HAE and on Madin-Darby canine kidney (MDCK) cells for virus isolation and passage.Cytopathic effects were observed daily after inoculation.Hemagglutination inhibition test was performed at every passage.Electron microscope was used to observe viral morphology.Viral genome was sequenced, followed by molecular evolutionary analysis.Results No progeny virus was isolated in MDCK cells, while a influenza A virus subtype H3N2 strain [A/Qinghai/178/2010(H3N2)] was isolated in HAE with a typical morphology and cytopathic effect of influenza A infection.The hemagglutination inhibition activity was 1∶16.Results of the molecular evolutionary analysis of viral genome showed that the influenza A virus (H3N2) strain was highly homologous to the A/Nanjing/1655/2010(H3N2) strain, which was isolated during the 2010 influenza pandemic in Nanjing.Conclusion HAE can be used for isolation and identification of virus from stale respiratory tract specimens.It is more sensitive than MDCK cells with regard to human influenza virus isolation.

Objective: To investigate the changes of gene expression in CD34+ hematopoietic stem and progenitor cells (HSPCs) under different growth environments. Methods: Umbilical cord blood mononuclear cells (UCB MNCs) were cultured in static and stirred systems. After 7 days of culture, CD34+ cells were isolated and total RNA was extracted. Gene expression patterns of CD34+ cells from fresh, static and stirred cultures were compared using differential display (DD). Results: 30 gene fragments displayed differential expression levels based on the conditions of DD. One of differentially expressed genes was identified as RAN, which is a member of oncogene RAS family. This gene may be associated with proliferation of hematopoietic cells. Conclusion: Different growth environments induced differential gene expression patterns of CD34+ HSPCs. These differentially expressed genes would give new insights into optimizing in vitro environments for expanding hematopoietic cells.

Culture environment is the key factor in the construction of dermal skin.It was investigated that the effects of the culture methods,including the static culture and spinner flask culture,and stir speeds on the cells proliferation,metabolism and distribution within collagenchitosan sponges.A higher cell density and specific growth rate was obtained with spinner flask culture versus static culture,especially,the 80 r/min spinner flask culture.The cell distribution in dermal substitutes from stirred culture system was more uniform than static culture,as well as that with increase of stir speeds in spinner flask.In summary,the spinner flasks culture with proper stir speed shows promise for the construction of dermal substitutes in vitro.

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.