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 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.

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.

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 report effects of microsurgical treatment for tumors in cervical spinal canal. Methods A total of 32 cases of tumors in cervical spinal canal had received microneurosurgery through a posternmedial approach. 22 cases had received reconstruction of cervical spinal canal stability. Results A total tumor resection was performed in 26 cases (81.25%),a subtotal tumor resection was conducted in 6 cases(18.75%). A cemplete recovery was achieved in 23 cases(71.88%), an improvement of symptoms was achieved in 6 cases(18.75%), and no improvement in 3 cases(9.38%), no death was encountered. Follow up observations were carded out in 30 cases for 3 months to 3 years. MRI examinatious 3 months after operation in 26 cases found no residual or recurrent tumor,recurrence was seen in 3cases of astrocytoma, 1 case was invariable. X-ray radiography in 26 cases showed good vertebral stability ,spinal deformation was found in 4 eases. Conclusion Microsurgery in early period play a key role in treatment of cervical spinal tumors. Reconstruction of spinal canal stability is important to patients.

Objective To study the factors affecting the prognosis of intracranial aneurysm operation for elderly patients. Methods Thirty elderly patients (≥60 years old) with intracranial aneurysm who were treated surgically were evaluated retrospectively. The factors that might affect the result of operation were analyzed by slng]e-factor and multi-factor. Result The factors that had significant correlation with the prognosis were Hunt-Hess grade,location of aneurysm,surgical timing (P =0.007,0.019,0.007).Conclusion The prognosis of intracranial aneurysm operation for elderly patients is affected by many factors,age is not a contraindication to aneurysm operation,early aneurysm operation treatment is suggested for elderly patients.

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.