AIM: To identify and quantitative analysis gentiopicroside content in the extract of Violent gentian of Tibetan herbal medicine. METHODS: Application of HPLC-the electricity sprays fog-mass spectroscopy/mass spectroscopy(HPLC-ESI-MS/MS) and the technique of LC-DAD-UV,by contrast with the reservation time of the authentic gentiopicroside,the ultraviolet absorbed spectrum,the molecule~+,quickly identified the gentiopicroside in the extract of violent gentian.As to it's quantitative analyis we adopted the capillary HPLC methods,with the weight of authentic gentiopicroside for abscissa. RESULTS: MS showed that the methanol extract of violet gentian mainly had five peaks.the compound structure of first peak was gentiopicroside,molecule~+ 356.9,result showed that gentiopicroside in gentian was 1.97%. CONCLUSION: Gentiopicroside is the mainly component of violet Gentia.

Objective To establish the quality control ofBaiduyin syrup.Methods The TLC was used to identity Radix paeoniae rubra, Radix Scutellariae. The quantitative determination of baicalin and buddleoside was completed by HPLC.Results The spots on TLC plates were distinct and high resolution. Compared with the negative samples, the contrast medicinal materials or control products showed that there were no spots of the same color in the corresponding position. The linear ranges of baicalin and buddleoside were 0.2179-2.1790 μg (r2=0.999 9), 0.1319-1.3190 μg (r2=0.999 7). TheRSD were 1.51% and 2.01%. Conclusions The established quality control method is simple, accurate and reproducible, which can be used for the quality control ofBaiduyin syrup.

Objective To establish the quality standard of Fufang-Shiwu-Zhixue powder. Methods The microscopical identification was adopted to analyze charred typha pollen and cuttlebone. TLC was used to identity rhubarb and radix notoginseng. UV-HPLC was used to determine the contents of notoginsenosides R1, ginsenosides Rg 1, ginsenosides Rb1. Results Microscopic identifications shower the characteristics of harred typha pollen and cuttlebone. The identified characteristics of rhubarb and radix notoginseng by TLC were distinct and the spots were clear. Notoginsenosides R1, ginsenosides Rg 1, ginsenosides Rb1 showed good linearity in the range of 0.16-1.58 μg (r=0.9998), 0.58-5.81 μg (r=0.9999), 0.33-3.29 μg (r=1.0000), respervtively.The average recoveries were 98.51% (RSD=1.81%), 97.80% (RSD=2.04%), 98.22%(RSD=1.45%). Conclusions The method is accurate, simple and repeatable, which could be used for the quality control of Fufang-Shiwu-Zhixue powder.

Objective To establish the quality standard ofQizhu-Fuzheng Yin, and to conduct a preliminary study on its stability.Methods Corydalis tuber and licorice were identified by TLC. The content of astragaloside was determined by UPLC-ELSD. The initial stability was studied by accelerated test method. Results The spots on TLC plates were clear without interference in the blank reference. The response of astragaloside was linear in the ranges of 36.5-365.0 μg/ml (r2=0.999 2), and the average recovery was 102.8 %, and theRSD was 2.4%. After 1, 2, 3, 6 months tests, the average contents of 3 batches astragaloside were 61.6, 60.4, 60.6μg/ml.ConclusionQizhu-Fuzheng Yin was simple preparation, quality control and stability.

BACKGROUND: Many scholars have paid attentions on neural stem cells (NSCs), which are regarded as the cell source to repair the injured nervous tissues. And it is becoming a key point that NSCs will be obtained in an effective way.OBJECTIVE: To observe the characteristics of isolation, proliferation and differentiation of in vitro NSCs from subventricular zone (SVZ) and hippocampus of neonatal rats.DESIGN: Single-sample trial.SETTING: Department of Neurosurgery in the Third Affiliated Hospital of Sun Yat-sen University.MATERIALS: Ten SD rats aged 3 days, of either gender, were offered from the Experimental Animal Center of Sun Yat-sen University. Nestin antibody (rabbit-anti-rat), neurofilament (NF-200) antibody (rabbit-anti-rat), and glial fibrillary acidic protein (GFAP) antibody (mice-anti-rat) were all purchased from Sigma Company.METHODS: The experiment was carried out in the Laboratory of Histology and Embryology, Basic Medical College of Sun Yat-sen University from September to December in 2006. According to the ethical requirement, the serum-free DMEM/F12 medium containing basic fibroblast growth factor and epidermal growth factor was utilized to isolate and incubate NSCs from SVZ and hippocampus of neonatal rats. Then the proliferation and differentiation of NSCs in vitro were observed.MAIN OUTCOME MEASURES: Fluorescence immunocytochemistry was applied to detect the antigen of nestin expressed from NSCs, NF expressed from neuron cells and GFAP expressed from astrocytes. The differentiation of NSCs was identified.RESULTS: The cells isolated from SVZ and hippocampus of neonatal rats possessed the ability of proliferation and self-renewal. Through inverted microscope, nestin positive cells could be found in the cell clone sphere at passage 20 of cells. Cells induced to differentiate could express antigen of NF and GFAP by the attached cell clone sphere.CONCLUSION: The NSCs successfully isolated from SVZ and hippocampi of neonatal rats possess the ability of self-renewal, proliferation, and differentiation into neuron-like cells and astrocytes.

OBJECTIVE:To determine the contents of cholalic acid and hyodeoxycholic acid in Qingkailing suppository by HPLC-ELSD(high performance liquid chromatography with evaporative light scattering detection). METHODS: The analytical column was Diamonsil C18(250 mm?4.6 mm, 5 ?m) and the mobile phase consisted of 0.1% formic acid-acetonitrile(gradient elution) with the temperature of the drift tube of the light scattering detector set at 110 ℃ and the flow rate of the nitrogen gas at 2.0 L?min-1. RESULTS: The linear ranges of cholalic acid and hyodeoxycholic acid were 0.59～7.4 ?g(r=0.999 6)and 0.38～4.8 ?g(r=0.999 7),respectively,with their average recoveries at 99.20%(RSD=1.4%,n=6)and 98.57%(RSD=1.8%,n=6), respectively. CONCLUSION: The method is simple, accurate and well-separated, and it is applicable for the quality control of Qingkailing suppository.

AIM: To observe the effect of receptor-interacting protein 2 (Rip2) overexpression on human pan-creatic cancer cell line Panc-1.METHODS: pEGFP-C2 and pEGFP-Rip2 plasmids were respectively transfected into the Panc-1 cells using JetPRIME reagent.The cells were divided into control group, pEGFP-C2 group and pEGFP-Rip2 group. The apoptosis in the cells was detected 48 h after transfection by flow cytometry.Rip2 level and the expression of apoptosis-related proteins, Bax, cytoplasmic cytochrome c ( Cyt-c) and Bcl-2, were analyzed by Western blot.The activity of caspase-3 was measured by colorimetric method.RESULTS: Rip2 protein expression significantly increased in the cells transfected with control and pEGFP-C2 plasmids.The apoptotic rate in pEGFP-Rip2 group was higher than that in control group and pEGFP-C2 group, whereas no significant difference of apoptotic rate was observed between control group and pEGFP-C2 group.The protein expression of Bax and cytoplasmic Cyt-c was remarkably increased and the protein expression of Bcl-2 was obviously decreased in pEGFP-Rip2 group as compared with control group and pEGFP-C2 group.The activity of caspase-3 in pEGFP-Rip2 group was obviously increased as compared with control group and pEGFP-C2 group.CON-CLUSION: Overexpression of Rip2 is able to induce apoptosis in the Panc-1 cells, and the mechanism may be related to the up-regulation of Bax and cytoplasmic Cyt-c protein expression, down-regulation of Bcl-2 protein expression and en-hancement of caspase-3 activity, thus activating intrinsic apoptotic pathway.

Objective To summary the microsurgery clinical experience of 21 patients with cerebellopontine angle tumor by the help of three dimensional individual digital anatomy. And to evaluate the value of three dimensional individual anatomy in the treatment of tumors in cerebellopontine angle. Methods Between January 2011 and November 2011,21 patients with various cerebellopontine angle tumor,managed at the Third Affiliated Hospital of Sun Yat-Sen University, underwent CTA scan, and reconstruct the local anatomy by 3D view software. According to the individual anatomical model, the microsurgery program by restrosig moid approach was developed. Results All patients had reposition of the bone flap at original site after craniectomy during the same operative setting mentioned above with retrosigmoid approach. No complication was noted.Patients did not have any delayed postcraniectomy pain at operation site.Postoperative computed tomography of the skull showed good healing and shaping of the suboccipital bone at the surgical region. Conclusion With the help of three dimensional individual anatomy, the microsurgery of cerebellopontine angle tumor underwent less postoperative complications.This study provides a safe and effective individualized microsurgical methods by restrosig moid approach.

The oral colon-specific drug delivery system (OCDDS) has gained more attention from investigators in recent years since it can increase local drug concentration and reduce dosage and side effects.The type of colonspecific drug delivery system consists of enzyme dependent,pH dependent,time dependent,pressure dependent system and CODEDSTM.The development of many new materials and technology is very important for the preparation of new type of precise positioning colon-specific preparation.This article summarizes the advances in excipients and technique for oral colon-specific drug delivery system in recent years.It may provide a reference and a basis for the researchers concerned.

AIM: To observe the effects of vascular endothelial growth factor(VEGF) on the proliferation and differentiation of neural stem cells(NSCs) of rats in vitro.METHODS: NSCs isolated from the hippocampal gyrus of SD rats were primary cultured and subcultured,and then divided into two groups:(1) the cells in VEGF group were treated with 150 ?g/L VEGF in the culture system,and VEGF was removed at the 7 th day;(2) control group(without VEGF treatment).The cellular morphology of two groups was observed by contrast phase microscope.Nestin and NF-200 expressing cells were detected via immunofluorescence method.The percentages of the immunostaining positive cells in each group at the 7 th day and at the 11 th day were determined.RESULTS: At the 7 th day,the percentage of nestin positive cells in VEGF group was 52.19%?7.95%,vs 29.26%?4.12% in control group(P