Objective To obtain highly purified oligodendrocyte precursor lineage cells in vitro and make identification.Methods The oligodendrocyte precursors were separated from astrocyte by orbital shaker and further purified by differential adhesion,and finally cultured in chemically defined serum-free medium,with appended neurotrophin 2(N2),platelet-derived growth factor(PDGF),basic fibroblast growth factor(bFGF).Immunofluorescence assay was applied to identify the separated cells with A2B5,O4,O1 and glial fibrillary acidic protein(GFAP) antibodies.Results Over 95% of cultured oligodendrocyte precursor cells were obtained.The oligodendrocyte progenitors were A2B5 and O4 positive,immature oligodendrocytes were O4 and O1 positive while GFAP were negative.Conclusions Separation and purification by shaking and differential adhesion and chemically defined medium are suitable and effective to obtain highly purified oligodendrocyte precursor cells.Cell output will increase notabily and rest in immature phase by appending both N2,PDGF and bFGF.

Sodium houttuyfonate (SH) is a derivative of effective component of a Chinese material medica, Houttuynia cordata, which is applied in anti-infection of microorganism. But, the antimicrobial mechanisms of SH still remain unclear. Here, we firstly discovered that SH effectively inhibits the three types of virulence related motility of.Pseudomonas aeruginosa, i.e., swimming, twitching and swarming. The plate assay results showed that the inhibitory action of SH against swimming and twitching in 24 h and swarming in 48 h is dose-dependent; and bacteria nearly lost all of the motile activities under the concentration of 1 x minimum inhibitory concentration (MIC) (512 mg x L(-1) same as azithromycin positive group (1 x MIC, 16 mg x L(-1)). Furthermore, we found that the expression of structural gene flgB and pilG is down-regulated by SH, which implies that inhibitory mechanism of SH against motility of P. aeruginosa may be due to the inhibition of flagella and pili bioformation of P. aeruginosa by SR Therefore, our presented results firstly demonstrate that SH effectively inhibits the motility activities of P. aeruginosa, and suggest that SH could be a promising antipseudomonas agents in clinic.
Alkanes ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Biofilms ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fimbriae, Bacterial ; drug effects ; genetics ; metabolism ; Houttuynia ; chemistry ; Pseudomonas aeruginosa ; cytology ; drug effects ; genetics ; pathogenicity ; Sulfites ; pharmacology ; Virulence ; drug effects

Objective To detect the gene expression changes between urethra plates from hypospadias patients and foreskins from non-hypospadias patients by microarray and to investigate the underlying mechanisms of hypospadias.Methods Twelve hypospadias patients,aged 6-12 months (mean,8 months),were enrolled as the test group,including 5 moderate and 7 severe hypospadias patients.Six age-matched patients underwent circumcision were enrolled as controls.Samples from hypospaidas patient's urethra plates during hypospadias repair and samples from the foreskins during circumcision were obtained and processed into Tri-Reagent immediately for RNA extraction.Oligonucleotide expression microarrays were used to detect genes expression changes in tissues from patients with and without hypospadias.This microarray analysis incorporated 22 000 genes.The intensity of all genes present was analyzed by one-way ANOVA (P＜0.01) and Tukey's test.Four estrogen responsive genes,CYR61,CTGF,ATF3 and GADD45β,were tested by RT-PCR in 8 controls,8 moderate hypospadias and 8 severe hypospadias as well.Results Ninty-four genes were detected differentially expressed in hypospadias patients compared with phimosis patients.There were 47 genes upregulated in moderate hypospadias compared with controls(P＜0.01),68 genes up-regulated in severe hypospadias compared with controls(P＜0.001),17 genes up-regulated in severe hypospadias compared with moderate hypospadias(P＞0.05).These genes were involved in different cell functions such as growth regulation and signal transduction.CYR61,CTGF,ATF3 and GADD45β,known to be estrogen responsive or to interact with estrogen receptor were found up-regulated in microarray and the up-regulations were confirmed by RT-PCR.Conclusions The up-regulated genes contribute to the development of hypospadias.Up-regulation of estrogen responsive genes may play important roles in the development of hypospadias.

Objective To investigate the effects of different intensity of wide band noise on acoustic stapedius reflex amplitude in sound field. Methods Acoustical reflex amplitude(at ART+10 dB)were obtained at three fre-quencies (1,2, 4 kHz) from 80 normally hearing subjects with wide band noise at 30,40,50 and 60 dB SL in sound field. Results Acoustical reflex amplitude was significantly decreased in the presence of wide band noise (P 0. 01). The most significant decrease of acoustic reflex amplitude could be observed in the present of 40 dB SL wide band noise. In comparison with the 1 kHz group, significant decrease was found in 2 kHz group and 4 kHz group(P <0. 01),but there was no difference between the 2 kHz group and 4 kHz group(P>0.05). Conclusion Since the decrease of the acoustic stapedius reflex amplitude is mediated by the auditory efferent system in sound field, in clini-cal practice the function of the auditory efferents can be evaluated by testing the change of acoustic stapedius reflex amplitude with the stimulation of the wide band noise in sound field.

OBJECTIVETo research the effects of Guizhi Fuling capsule on sex hormones levels in blood serum and breast issue morphology of hyperplasia of mammary glands model rats.

METHODThe unpregnancy SD rat models of hyperplasia of mammary glands were established by injecting 0.5 mg x kg(-1) benzoate estradiol. After five weeks doses,the effects of Guizhi Fuling capsule 2.0, 1.0, 0.5 g x kg(-1) and Rupixiao tablet 0.5 g x kg(-1) on the changes of papilla diameter, height and breast issue morphology of the naimal models were explored, and sex hormones levels in blood serum were measured.

RESULTGuizhi Fuling capsule can inhibitnipple swell, improve breast tissue morphology pathological profiles of the animal models, and decrease oestradiol (E2) level and increase progesterone (P) level in blood serum.

CONCLUSIONThese results suggested that Guizhi Fuling capsule could, improve mammary gland pathological profiles. Regulating sex hormone levels may be its important mechanism for treatment of hyperplasia of mammary glands.

Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadal Steroid Hormones ; blood ; Hyperplasia ; Mammary Glands, Animal ; drug effects ; pathology ; Rats

OBJECTIVETo establish an analytical method for the fingerprint of triterpenoid constituents of Poria by HPLC and compare the fingerprints of different medicinal parts of Poria in order to provide basis for controlling Poria quality.

METHODThe HPLC chromatographic conditions were Waters Symmetry C18 column (4.6 mm x 250 mm, 5 μm), 0.1% phosphoric acid (A) and acetonitrile (B) as gradient mobile phases, flow rate being 1.0 mL x min(-1), column temperature at 30 degrees C, The detection wavelength was set at 210 nm; The cluster analysis was carried on by SPSS 15.0.

RESULTThe HPLC fingerprints of triterpenoid constituents of Poria were set up. There were 16 common peaks in different medicinal parts. The results of method validation met technical requirement of fingerprints; Triterpenoid constituents in White Poria and Poria cum Radix Pini were different from Poria. The content of pachymic acid was the highest in Poria. The effect of habitat on the quality was no obvious difference.

CONCLUSIONThe method is stable, reliable, reproducible, and can be used as an effective means of Poria quality evaluation.

Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Poria ; chemistry ; Triterpenes ; analysis

A novel white-rot fungus AH28-2,which was isolated from 224 fungi samples,ability to produce effectively laccase by induction.Several factors influencing laccase production were investigated.The optimum conditions were as follows:the 300mL Erlenmeyer flask containing 150mL of liquid medium was inoculated with 7.5mL of mycelial fragments and the medium was supplied with lignin at a concentration of 0.1%(initial pH8.5).The cultures was incubated at 28℃ on rotary shaker(150r/min) for 4～5 days.The maximum enzyme activity was 20184IU/L.

OBJECTIVETo present the experience in using the Snodgrass technique, Duckett repair and bladder mucosa grafting for hypospadias.

METHODSWe retrospectively reviewed 251 cases of middle and posterior hypospadias treated by the Snodgrass technique, Duckett repair and bladder mucosa grafting from February 1997 to December 2008.

RESULTSThe success rates of the Snodgrass technique, Duckett repair and bladder mucosa grafting were 80.3% (53/66) , 76% (19/25) and 78.9% (15/19) for middle hypospadias and 68. 3% (41/60) , 63.6% (14/22) and 88. 1% (52/59) for the posterior type without statistical significant differences (all P > 0.05). Bladder mucosa grafting showed a significantly higher success rate than the other two procedures in either one-stage or two-stage surgical repair of posterior hypospadias (P < 0.05). Duckett repair achieved a significantly higher rate of success in children under 14 years than in the older ones (P < 0.05).

CONCLUSIONOf the three surgical options, bladder mucosa grafting is the most suitable for posterior hypospadias, and Duckett repair is recommended for children under 14 years of age.

Adolescent ; Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Mucous Membrane ; surgery ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Urinary Bladder ; surgery

OBJECTIVEThe purpose of this study is to study the sealing ability and the furcal appearance of repairing subpulpal wall perforation with resinous inlay.

METHODSFifty newly extracted human molars were randomly divided into three experiment groups (group A, group B, group C, 15 teeth each) and one control group (5 teeth). In experiment groups, perforations were made perpendicularly to the center of the pulp chamber floor. Perforations of group A and B were repaired with resinous inlay and sealed by AH Plus sealer and luting glass-ionomer, respectively. Perforations of group C were directly repaired using light-cure composite resin. Perforations were not made in five teeth of control group. The furcal appearances were evaluated under stereomicroscope after repairing. Microleakage was measured by glucose oxidase detection.

RESULTSThe fineness rate of furcal appearances with resinous inlay repairing were 83.3%, while the fineness rate of furcal appearances with light-cure composite resin directly repairing were 46.7%. There were statistics difference between resinous inlay repairing and light-cure composite resin directly repairing (P<0.05). There were statistics difference among the daily microleakage of three experiment groups, group A

CONCLUSIONUsing resinous inlay to repair the subpulpal wall perforation can improve the sealing effect and avoid material overextension. AH Plus can be used as perforation sealant because of its better sealing ability.

Bicuspid ; Composite Resins ; Dental Leakage ; Dental Pulp Cavity ; Glass Ionomer Cements ; Humans ; Inlays ; Molar

OBJECTIVEto explore the value of in vivo dynamic monitoring of abdominal aortic atherosclerosis (AS) by high field magnetic resonance (MR) imaging (MRI) in apoE-/- mice fed a high fat diet or infused with angiotensin.

METHODShigh fat diet or angiotensin II infusion was applied to apoE-/- mice for establishment of abdominal aortic atherosclerosis model. Abdominal aorta MRI was performed at 3 time points (baseline, 3 and 6 months) in 13 high fat diet fed apoE-/- mice aged 10-12 months and 3 wild-type control mice; 10 apoE-/- mice aged 6 months were infused with angiotensin II (1000 or 500 ng × kg(-1)× min(-1), n = 5 each) or saline for 14 d through Osmotic minipump. The abdominal aortic artery MRI was performed at baseline and 14 d after infusion. Black blood sequences of FLASH T1 weighted images and Proton density weighted-T2 weighted dual echo images were obtained. At each observation time post MRI, mice (n = 3, 5 and 5 for high fat diet group and n = 5 and 5 for angiotensin II infusion group) were sacrificed for pathological examination of the abdominal artery.

RESULTS(1) the abdominal aorta atherosclerosis was identified in both high fat diet and angiotensin II treated apoE-/- mice but in WT controls. Lesion progression was documented in high fat diet fed apoE-/- mice characterized by significantly increased vessel wall (a marker of atherosclerotic burden, F = 29.94, P < 0.05) and gradually increased plaque signal in PDW and T2W images. Results derived from MRI corresponded histopathology findings in high fat diet fed apoE-/- mice (correlative coefficient = 0.84, 0.95, 0.90, P < 0.05, respectively). Both MRI and histology showed increased lipid composition and decreased fibrotic composition in these mice. (2) The vessel wall area increased significantly [(1.21 ± 0.21) mm(2) vs. (2.65 ± 0.48) mm(2), P < 0.05] and the abdominal aortic dissection aneurysms was identified in apoE-/- mice infused with high angiotensin II. The vessel wall area also increased [(0.85 ± 0.11) mm(2) vs. (1.01 ± 0.17) mm(2), P < 0.05] in low angiotensin II infused apoE-/- mice and the coefficient between MR and histopathology is 0.934.

CONCLUSIONabdominal aortic unstable plaque model could be established by both high fat diet and angiotensin II infusion in apoE mice, angiotensin II infusion can transiently accelerate the progression of AS and can induce abdominal aortic dissection. Serial MR black blood sequences could demonstrate the development and progression of atherosclerosis in mouse abdominal aorta with excellent agreement to histopathology finding in terms of atherosclerotic burden and plaque composition. Thus, MRI appears to be a useful tool for in vivo AS plaque dynamic monitoring in mice.

Angiotensin II ; administration & dosage ; Animals ; Aorta, Abdominal ; Apolipoproteins E ; Arteriosclerosis ; Diet ; Dietary Fats ; administration & dosage ; Disease Models, Animal ; Magnetic Resonance Imaging ; methods ; Male ; Mice ; Mice, Knockout