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.

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.

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.

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

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.

Objective To evaluate the efficacy of bisphosphonate thera-pies regarding the treatment of vertebraland non vertebral-non hip frac-tures in postmenopausal women with osteoporosis.Methods We searched systematic reviews on the databases in Cochrane Library , PubMed, SCI, CNKI, VIP, CBM and Wangfang database, as well as hand searching.The reviews were identified with a systematic literature search.Then the data were extracted by standardized forms, and the methodological qualities of the reviews were assessed by the AMSTAR tool.The endpoints of interest were vertebral and non -vertebral frac-tures.The Meta-analysis was evaluated by RevMan 5.3 software.Re-sults of all reviews were analyzed byindirect comparison.Results A to-tal of 12 systematic reviews were included.There were six bisphopho-nates in these reviews ( including alendronate, ibandrinate, etidronate, risedronate, pamidronate, and zoledronic acid).All the reviews were in high quality.The results of Meta-analysis showed that there was statisti-cally significant by five bisphosphonates, expect for pamidronate.So there were five bisphosphonates ( alendronate, ibandrinate, etidronate, risedronate, and zoledronic acid ) analyzed with indirect comparion.In vertebral fractures of all bisphophonates compared, zoledronic acidshowed a relative risk ( RR ) of 0.49 relative to placebo, and RRs were 0.49, 0.53, 0.51, 0.74 relative to risedronate, ibandronate, alendronate, etidronate.Regarding non-vertebral fractures, RRs of zoledronic acid relative to placebo, risedronate, alendronate, ibandronate, etidronate were 0.72, 0.71, 0.87, 0.89, and 0.90, respectively.Risedronate showed the greatest reduction in any fractures, followed by zoledronic acid.Conclusion In summary, zoledronic acid has the best effectiveness on the treatment of osteoporosis among the five bisphosphonate.

Pharmaceutical cocrystals is an advanced technology to improve the physicochemical and biological properties of drugs. However, there are few studies on the in vivo metabolism of pharmaceutical cocrystals. In this study, the pharmacokinetics of wogonin cocrystal in normal rats was further studied on the basis of the previous preparation of wogonin-aloperine cocrystal. Firstly, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for simultaneous determination of wogonin and its metabolite wogonoside in rat plasma, and to investigate the methodology. The method was applied to the pharmacokinetic study of wogonin-aloperine cocrystal (Wog-Alop) in rats. The results showed that wogonin and its metabolite wogonoside had a good linear relationship in the range of 1-800 ng·mL^-1, and the precision, accuracy, matrix effect and stability in this range met the requirements of biological analysis. Compared with direct administration of wogonin, the C_max of wogonin and its metabolite in rats increased to 7.44-fold and 9.15-fold, AUC_0-t increased to 1.67-fold and 3.72-fold, and oral bioavailability of wogonin increased to 187.66% after cocrystal administration. Wog-Alop cocrystal can significantly improve the C_max, AUC, and oral bioavailability of wogonin and its metabolite, which provides a new perspective for the clinical application of wogonin. This study was approved by the Experimental Animal Ethics Review Committee of Beijing University of Chinese Medicine (approval number: BUCM-2023032307-1148).

OBJECTIVETo explore the effects of oxymatrine (OMT) on NF-kappaB and other cell factors in rat lung tissue with septic shock.

METHODFifty-six male SD rats were randomly divided into 7 groups: sham operation group, OMT control group, model (CLP) group, CLP + OMT high, middle, low-dose group, positive control group. Changes in NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue were determined by immunohistochemical method, tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in pulmonary tissue were determined by radioimmunoassay.

RESULTOMT could decrease significantly the NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue (P < 0.05), TNF-alpha and IL-6 levels in pulmonary tissue homogenate decreased markedly (P < 0.05). OMT could elevate the content of PaO2, SaO2, decrease the content of PaCO2, HCO3- and decrease the ratio between wet weight of the lung and dry weight of the lung and the PWI.

CONCLUSIONOMT can inhibit NF-kappaB-inducing kinase (NIK), NF-kappaB activity and reduce the expression of proinflammatory factor (TNF-alpha, IL-6) and antagonize the lung injury in a rat model of septic shock.

Alkaloids ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Gene Expression Regulation ; drug effects ; I-kappa B Proteins ; metabolism ; Immunohistochemistry ; Interleukin-6 ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Quinolizines ; pharmacology ; Radioimmunoassay ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism