To study the in situ intestinal absorption of five oligosaccharides contained in Morinda officinalis How. (sucrose, kestose, nystose, 1F-Fructofuranosyinystose and Bajijiasu). The absorption of the five oligosaccharides in small intestine (duodenum, jejunum and ileum) and colon of rats and their contents were investigated by using in situ single-pass perfusion model and HPLC-ELSD. The effects of drug concentration, pH in perfusate and P-glycoprotein inhibitor on the intestinal absorption were investigated to define the intestinal absorption mechanism of the five oligosaccharides in rats. According to the results, all of the five oligosaccharides were absorbed in the whole intestine, and their absorption rates were affected by the pH of the perfusion solution, drug concentration and intestinal segments. Verapamil Hydrochloride could significantly increase the absorptive amount of sucrose and Bajijiasu, suggesting sucrose and Bajijiasu are P-gp's substrate. The five oligosaccharides are absorbed mainly through passive diffusion in the intestinal segments, without saturated absorption. They are absorbed well in all intestines and mainly in duodenum and jejunum.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Intestinal Absorption ; Intestine, Small ; metabolism ; Male ; Morinda ; chemistry ; Oligosaccharides ; chemistry ; pharmacokinetics ; Perfusion ; Rats ; Rats, Sprague-Dawley

Objective To investigate the CT spectral imaging features of pancreatic serous oligocystic adenoma and mucinous cystic neoplasms and to assess the value of spectral CT in differentiating between pancreatic serous oligocystic adenoma and mucinous cystic neoplasms. Methods From Feb.2010 to Dec. 2010, 27 patients with cystic neoplasms of the pancreas (group one with 15 serous oligocystic adenomas and group two with 12 mucinous cystic neoplasms) underwent dual-phase CT spectral imaging followed by surgery. Quantitative values (age, tumor size, CT value change as function of photon energy,effective-Z, iodine-water concentration, and calcium-water concentration) were compared with independent samples t test and Mann-Whitney test and non-quantitative parameters (gender, symptom, and tumor location) were compared with Chi-square test (Fisher exact). The parameters with significant differences between two groups were analyzed further and the performance of multiple parameters for joint differential diagnosis was evaluated with discriminant analysis. Results Compared to patients with mucinous cystic neoplasms, patients with serous oligocystic adenoma had younger age, lower frequency of being symptomatic and smaller tumor size. The CT values on 40 keV to 60 keV( with 10 keV increment) in late arterial phase [(36±13)HU vs. (62±23)HU, (26 ±8)HU vs. (40±15)HU, and (19±6)HU vs. (27±10)HU respectively] and 40 keV to 50 keV (with 10 keV increment) in portal venous phase [ (43 ± 14 )HU vs.(61 ±25)HU and (30 -10)HU vs. (40 ± 16)HU respectively], effective-Z (late arterial phase 7.80 ± 0. 16 vs. 8.05 ± 0. 21, and portal venous phase 7. 87 ± 0. 15 vs 8.02 ± 0. 22 ), concentration of calcium (water) [late arterial phase (5 ±3) g/L vs. (11 ±4) g/L, t= -3.836, P=0.001 and portal venous phase (7 ± 3 ) g/L vs. ( 10 ± 5 ) g/L, t = - 2.071, P = 0. 049 ] and iodine (water) [ late arterial phase (0.38 ±0.24) g/L vs. (0.78 ±0.32) g/L, t = -3.755, P=0.001 and portal venous phase (0.48 ± 0. 24) g/L vs. (0. 72 ± 0. 34 ) g/L, t = - 2. 161, P = 0. 041 ] were lower in serous oligocystic adenoma than those in mucinous cystic neoplasms. In discriminant analysis, multiple parameters [ age, symptom,tumor size, CT values on 40 keV to 50 keV, effective-Z, concentration of iodine (water) in late arterial phase and concentration of calcium (water) in portal venous phase] showed high accuracy (100%, 27/27 )of joint diagnosis between serous oligocystic adenoma (100%, 15/15 ) and mucinous cystic neoplasms (100%, 12/12). Conclusions The serous oligocystic adenoma and mucinous cystic neoplasms had distinct characteristic findings on CT spectral imaging. CT spectral imaging is highly accurate in the differential diagnosis between serous oligocystic adenoma and mucinous cystic neoplasms.

Objective To evaluate the efficacy and safety of topical retinoids in the treatment of acne vulgaris compared with placebo, antibiotics, benzoyl peroxide and sulfur preparation. Methods According to the Cochrane reviewer′s handbook, randomized controlled clinical trials were selected for the systematic review. Results Up to 2002, 15 clinical trials (2,439 patients) that met the inclusion criteria were selected. There were four clinical trials which showed that topical retinoids were more effective than that of placebo (RR=1.87, and 95% CI: 1.13 ~ 3.11),especially for noninflammatory lesions (RR=12.70,and 95% CI : 4.09 ~ 39.40). There were 3 clinical trials which showed that topical retinoids had better efficacy than that of sulfur preparations (RR=1.75, and 95% CI: 1.42 ~ 2.16). For 7 clinical trials of retinoids compared with benzoyl peroxide, and 3 clinical trials of retinoids compared with antibiotics, no conclusion could be drawn. All the clinical trials showed that there were local side effects, including erythema, and scaling etc in the patients using topical retinoids, but no systematic side effects were observed, however, pregnant women had to be very cautious. Conclusions Topical retinoids are effective for acne vulgaris, and has better efficacy than sulfur preparation does, but there is not enough evidence to clarify that the efficacy of topical retinoids is better than that of benzoyl peroxide and antibiotics.

OBJECTIVETo evaluate the effect of early application of recombinant human erythropoietin (rhEPO) on white matter development in preterm infants using fractional anisotropy (FA) of magnetic resonance diffusion tensor imaging (DTI).

METHODSA total of 81 preterm infants with gestational age ≤32 weeks, birth weight <1 500 g, and hospitalization within 24 hours after birth were randomly divided into rhEPO group (42 infants) and control group (39 infants). The infants in the rhEPO group were administered rhEPO, while those in the control group were given the same volume of normal saline. The preterm infants of both groups took examinations of head magnetic resonance imaging, diffusion-weighted imaging, and DTI at the corrected gestational age of 35-37 weeks. FA was calculated for the regions of interest in both groups.

RESULTSThere was no significant difference in the incidence of intracranial hemorrhage, periventricular leukomalacia, focal cerebral white matter damage (CWMD), and extensive CWMD between rhEPO and control groups (P>0.05). Compared with the control group, the rhEPO group showed higher FA values at the posterior limb of the internal capsule, the splenium of the corpus callosum, frontal white matter, and occipital white matter (P<0.05). There was no significant difference in FA values at the parietal white matter, thalamus, lenticular nucleus, and caudate nucleus between the two groups (P>0.05).

CONCLUSIONSEarly application of rhEPO has a neuroprotective effect on white matter development in preterm infants.

Diffusion Tensor Imaging ; Erythropoietin ; pharmacology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Neuroprotective Agents ; pharmacology ; Recombinant Proteins ; pharmacology ; White Matter ; drug effects ; growth & development

OBJECTIVETo determine determinate five oligosaccharides, namely sucrose, 1-kestose, nystose, 1F-fructofurano-syinystose, bajijiasu contained in Morinda officinalis with an HILIC-ELSDI) method.

METHODWaters XBridge Amide (4.6 mm x 150 mm, 3.5 microm) hilic column was adopted for gradient elution, with acetonitrile (A) and 0.2% triethylamine (B) as the mobile phase. The column temperature was set at 40 degrees C, with the flow rate of 0.8 mL x min. Waters 2424 evaporative light scattering detector (ESLD) was used as detector, with the gas flow of 275.79 kPa and drift tube temperature of 90 degrees C.

RESULTThe detection range for the five oligosaccharides were 2.128-21.28 microg for sucrose (r = 0.999 3), 1.864-18.64 microg for 1-kestose (r = 0.999 6), 1.92-19.2 microg for nystose (r = 0.999 8), 1.912-19. 12 microg for 1F-fructofuranosyinystose (r = 0.999 5), 2.368-23.68 microg for bajijiasu (r = 0.999 4), respectively. The recovery of the five oligosaccharides ranged between 92.81%-102.8% (n = 6).

CONCLUSIONThe method is so simple, accurate and highly reproducible that it can be used as an analytical method for effective evaluation of the quality of M. officinalis herbs.

Chromatography, Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Morinda ; chemistry ; Oligosaccharides ; analysis ; Scattering, Radiation

OBJECTIVETo investigate the specific antitumor immune response induced by idiotype protein (Id)-pulsed dendritic cells (DC) in vitro.

METHODSDC was generated from peripheral blood monocytes of the multiple myeloma (MM) patients using GM-CSF, IL-4, and TNF-alpha. The DCs were pulsed with idiotypic fragment, the F(ab')2 fragment of M protein from MM patient at the immature stage. The morphologic characteristics of the cells were observed with light and electron microscopes. The phenotypic features were analyzed with FACS, MTT assay was employed to evaluate the proliferation of autologous T cells and the inhibition rate of MM cells.

RESULTSDC precursors in peripheral blood could be induced to typical mature DC in medium containing GM-CSF, IL-4 and TNF-alpha. Mature DC with Id could increase the proliferation of the autologous T cells and activate naive T cells to become tumor specialized cytotoxic T lymphocytes (CTL). The CTL at different doses showed significant inhibition on or killing ability to autologous MM cells in vitro.

CONCLUSIONSIn a suitable cytokine environment, the DC precursors from peripheral blood of MM patients could be induced to functional DC, and vaccination of Id-pulsed DC could induce active antitumor immune response.

Adult ; Aged ; Antibodies, Anti-Idiotypic ; immunology ; Cancer Vaccines ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy, Active ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study the relationship between Nanog-promoted metastasis of breast cancer and ezrin(T567) phosphorylation, and explore the possible mechanism by which Nanog regulates ezrin(T567) phosphorylation.

METHODSA siRNA construct targeting Nanog was transfected in breast cancer cells to knock down Nanog expression, and the changes in the cell invasion was detected using Transwell assay. The expression levels of Nanog and PKC and the phosphorylation level of ezrin(T567) were detected using Western blotting and immunofluorescent staining; the protein interaction between PKCε and ezrin was assayed by co-immunoprecipitation and Western blotting.

RESULTSNanog knockdown significantly decreased the expression of PKCε protein, phosphorylation level of ezrin(T567) and the invasion ability of breast cancer cells. PKCε knockdown obviously decreased the phosphorylation level of ezrin(T567) in the cells, and PKCε and ezrin were co-immunoprecipitated.

CONCLUDIONSNanogcan can upregulate the expression of PKCε to promote the phosphorylation of ezrin(T567), which can be a new mechanism by which Nanog promotes tumor metastasis.

Blotting, Western ; Breast Neoplasms ; metabolism ; Cytoskeletal Proteins ; metabolism ; Gene Knockdown Techniques ; Homeodomain Proteins ; metabolism ; Humans ; Nanog Homeobox Protein ; Neoplasm Invasiveness ; Phosphorylation ; Protein Kinase C-epsilon ; metabolism ; RNA, Small Interfering ; Transfection ; Tumor Cells, Cultured ; Up-Regulation

OBJECTIVETo develop a multiplexed mutagenically separated PCR (MS-PCR) for single nucleotide polymorphism (SNP) loci typing in mitochondrial DNA coding regions and to study the applications in investigating the allele frequencies and haplotypes of four SNP loci in mitochondrial DNA coding regions in Chinese Chengdu Han population.

METHODSFour SNP loci C12705T, A8701G, G8584A and C10400T, two allele specific forward primer with 4 bases different in size and a common reverse primer were designed for SNP typing. The primers simultaneously were amplified in a single tube. The genotyping of SNPs was determined by the two allele specific fragments different in size after polyacrylamide gel and silver staining.

RESULTSThe different SNP loci comprised a single band with different size respectively. Typing results were completely consistent with those by direct sequencing. The allelic frequencies of C12705T, A8701G, G8584A and C10400T were 0.3813/0.6187, 0.4813/0.5187, 0.8250/0.1750 and 0.4938/0.5062 respectively. A total of 6 different haplotypes was identified and the genetic diversity reached 0.7137.

CONCLUSIONMultiplexed MS-PCR is a simple, rapid, accurate and efficient method for SNP typing, which will be very powerful for SNPs in the database establishing of mitochondrial DNA coding regions, the testing of forensic and population genetics research.

Base Sequence ; DNA, Mitochondrial ; chemistry ; genetics ; Gene Frequency ; Genetic Variation ; Genotype ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; genetics ; Sequence Analysis, DNA

OBJECTIVETo investigate the effect of precursor of nerve growth factor (proNGF) in promoting invasion of breast cancer cells and its relation with ezrin expression and phosphorylation of ezrin Thr567 and Tyr477.

METHODSHuman breast cancer cell lines MDA-MB-231 and MCF-7 were stimulated by gradient concentrations of proNGF (0, 2.5, 5 and 10 ng/mL) for 16 h, and the invasion of the cells was assessed with Transwell assay. The expression of ezrin and the phosphorylation of ezrin Thr567 and ezrin Tyr477 in the treated cells were examined by Western blotting. MDA-MB-231 cells were transfected with pEnter-His-ezrinY477F (a dominant negative mutant) to study the role of phosphrylation of ezrin Tyr477 in the invasion of breast cancer cell stimulated by proNGF.

RESULTSproNGF significantly promoted MDA-MB-231 and MCF-7 cell invasion in a concentration-dependent manner (P<0.05), and concentration- and time-dependently increased the phosphorylation of ezrin Tyr477 (P<0.05) without affecting the expression of ezrin or the phosphorylation of ezrin Thr567. The specific inhibitor of src, SKI-606, significantly inhibited the phosphorylation of ezrin Tyr477 induced by proNGF. Transfection with pEnter-His- ezrinY477F inhibited proNGF-induced invasion and phosphorylation of ezrin Tyr477 in MDA-MB-231 cells (P<0.05).

CONCLUSIONPhosphorylation of ezrin Tyr477 plays a critical role in the invasion of breast cancer cells stimulated by proNGF via proNGF/src/ezrin Tyr477 pathway.

Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cytoskeletal Proteins ; chemistry ; Humans ; MCF-7 Cells ; Neoplasm Invasiveness ; Nerve Growth Factor ; pharmacology ; Phosphorylation ; Signal Transduction ; Transfection ; Tyrosine

OBJECTIVETo investigate the anti-inflammatory effect of bone marrow stromal cells (MSCs) transfected with recombinant adenovirus-mediated ciliary neurotrophic factor (CNTF) gene in C57BL/6 mice with experimental allergic encephalomyelitis (EAE).

METHODSAn adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs (MSC-CNTF). After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-alpha (TNF-alpha), inteferon-gamma (IFN-gamma), interleukin-12P35 (IL-12P35), and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3+ T cell infiltration and TNF-alpha and IFN-gamma expressions in the lesions were also observed after the cell transplantation.

RESULTSCNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-alpha levels, ameliorated CD3+T cell infiltrations and lowered TNF-alpha expression in the lesions, while the levels of IFN-gamma underwent no significant changes.

CONCLUSIONTransplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-alpha and IFN-gamma levels and reduced inflammatory cells, CD3-positive cells and TNF-alpha expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.

Adenoviridae ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; metabolism ; Ciliary Neurotrophic Factor ; biosynthesis ; genetics ; therapeutic use ; Encephalomyelitis, Autoimmune, Experimental ; therapy ; Female ; Genetic Therapy ; Interferon-gamma ; blood ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Stromal Cells ; metabolism ; T-Lymphocytes ; immunology ; Transfection ; Tumor Necrosis Factor-alpha ; blood