OBJECTIVE: To establish an HPLC method for the determination of Matrine in Baishan tablets. METHODS: The analytical column was Aminopropyl (200 mm?4.6 mm, 10 ?m) with mobile phase consisted of acetonitrile-0.5% phosphate solution - ethanol(100∶8∶10) at a flow rate of 1.0 mL?min-1. The column temperature was set at 30 ℃ and the detection wavelength was set at 220 nm. RESULTS: The linear range of Matrine was 0.27～ 8.66 ?g(r=0.999 8) and the average recovery of Matrine was 99.52 %(RSD=0.91%,n=6). CONCLUSION: The method is simple, accurate and reproducible, and it is applicable for the quality control of Baishan tablets.

Carcinoma, Hepatocellular ; Humans ; Liver Neoplasms

BACKGROUND:Vascular endothelial growth factors (VEGF) can promote the division of endothelial cells and accelerate the growth of newborn vessels, whereas the expression and distribution of VEGF receptors (VEGFR) in spinal cord should be observed.OBJECTIVE: To observe the expression and distribution of VEGFR-1 and VEGFR-2 in rat spinal cord.DESIGN: A randomized controlled observation.SETTING: Department of Orthopaedic Surgery, the First Affiliated Hospital of Dalian Medical University; Department of Hand Surgery, Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Ten adult male Wistar rats of clean degree, weighing 180-200 g, were provided by the animal experimental center of Tongji Medical College, Huazhong University of Science and Technology. Immunohistochemical primary antibody was purchased from Santa Cruz Company; the second and third antibodies from Beijing Zhongshan Biotechnology Co., Ltd. Reverse transcription-polymerase chain reaction (RT-PCR) kit was the product of Promega,Trizol reagents were purchased from Invitrogen Company, and the VEGFR-1 and VEGFR-2 primers were designed by Beijing Aoke Company.METHODS: The experiment was carried out in the laboratory of Department of Hand Surgery of Wuhan Union Hospital from March to June in 2004. ① Detection of VEGFR expression in spinal cord anterior horn: The rats were anesthetized by 100 g/L chloral hydrate, spinal cord of lumbar 4-6 (L4-6) was fixed in fixation solution at 4 ℃ for overnight, then routine dehydration, hyalinization and paraffin embedding were performed, and serial sections of about 5 μm were prepared for observing the VEGFR expressions using immuniohistochemical staining. ② Detection of VEGFR mRNA expression in spinal cord: Five rats were selected, L4-6 spinal cord (50 mg) was removed and centrifugated, then the content of total RNA was determined with ultraviolet spectrophotometer. The synthetized cDNA was amplified with PCR, and the PCR products (10 μL) were treated with 20 g/L agarose gel electrophoresis, and stained with 0.1 mg/L ethidium bromide, the results were observed and recorded under ultraviolet lamp and photographed. The products were scanned and quantified with gel imaging analytical system to record the gray value of each band, the gray value of β-actin band was taken as 1,and those of the other objective fragments were compared with it to record the gray value ratio and analyze the expressions of the objective fragments.MAIN OUTCOMEMEASURES: ① Expression and distribution of VEGFR-1 and VEGFR-2 in rat spinal cord anterior horn;② Results of VEGFR mRNA expression in spinal cord.RESULTS: ① Both VEGFR-1 and VEGFR-2 were expressed in the microvessels of normal rat spinal cord tissue.Besides, VEGFR-2 mainly expressed in motor neurons, glial cells and the nerve fibers in surrounding white matter. ②The results of gel imaging analytical system showed that the VEGFR-2 content in normal spinal cord was obviously higher than VEGFR-1 (0.874±0.222, 0.486±0.181, P ＜ 0.05).CONCLUSION: VEGF promote the formation of microvessels through the combined effects of VEGFR-1 and VEGFR-2,and it plays the neurotrophic and neuroprotective role through VEGFR-2.

Hemoglobinuria, Paroxysmal ; metabolism ; Humans ; Membrane Proteins ; biosynthesis

Adult ; Aged ; Aged, 80 and over ; Anemia ; chemically induced ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Carboplatin ; administration & dosage ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; chemically induced ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Paclitaxel ; administration & dosage ; Remission Induction ; Survival Rate ; Thrombocytopenia ; chemically induced

Animals ; Female ; Herbicides ; toxicity ; Male ; Mice ; Mice, Inbred Strains ; Testis ; drug effects ; Thiocarbamates ; toxicity ; Thyroid Gland ; drug effects ; Weight Gain ; drug effects

This study is to investigate the effect of local phVEGF(165) injection on sciatic nerve regeneration in the rats and to search for a new way in the further treatment of peripheral nerve injuries. Forty-five adult male Wistar rats received a neurotomy to bilateral sciatic nerves, which were subsequently reconnected with 10/0 epineurial nylon sutures. The injured segments was locally injected with normal saline (group A), or 25 microg of phVEGF(165) (group B) or 50 microg phVEGF(165) (group C). Nerve conduction and regeneration were evaluated in terms of the histological changes, weight of gastrocnemius muscles, electrophysiology and morphometric results. Our study demonstrated that rats of group C showed the best results in terms of nerve regeneration, followed by group B and group A. Our findings suggested that local injection of phVEGF165 can facilitate nerve regeneration and promote functional recovery in a dose-dependent manner.

Objective To investigate the changes in CREB_1 proteins in five brain regions of rats with morphine addiction and withdrawal with the technique of immunohistochemistry. Methods Thirty six adult male Sprague-Dawley rats were randomly divided into six groups (n=6 for each), i.e. acute morphine dependent group, acute abstinence group, acute control group, chronic morphine dependent group, chronic abstinence group and chronic control group. Animals in dependent groups and abstinence groups were administered with morphine by intraperitoneal injection till morphine dependent models were established. The rats in abstinence groups withdrawal syndromes were induced with naloxone 5mg/kg for 30min. The rats in control groups were injected with saline. All rats were sacrificed by decapitation. The coronal sections of discrete brain regions (hypothalamus, nucleus accumbens, prefrontal cortex, locus coeruleus, hippocampus) were obtained. The relative concentrations of CREB_1 protein were determined with immunohistochemistry. Results In acute morphine dependent and abstinence groups, CREB_1 protein decreased significantly compared with the acute control group in locus coeruleus (P0.05). Conclusion The morphine-induced CREB_1 protein changes may reflect differential G protein-cAMP-CREB signal transduction pathways in morphine dependent and abstinence rats.

Objective To study the effects of continuous oxygenated blood microperfusion on rat hearts preservation.Methods Forty-five Wistar rats were randomly divided into 3 groups.The isolated rat hearts were continuously microperfused with oxygenated blood at 4℃in experimental group,those in control group microperfused with St.Thomas solution and those in solution group were preserved with St.Thomas solution.After preservation the hemodynamic indexes of rat hearts were measured using Langendorff system,including the left ventricular end-diastolic pressure (LVEDP),the left ventricular systolic pressure (LVSP),the left ventricular developed pressure (LVDP) and the rate of the left ventricular pressure change (?dp/dt),then the contents of adenosine triphosphate (ATP) were determined by high-performance liquid chromatography and myoeardium ul- trastructures were observed under an electric microscope.Results After 30 min perfusion of Langen- dorff system,the LVSP,LVDP and?dp/dt were (38,25?3.84) mm Hg,(32.54?4.01) mm Hg and (1080?123) mm Hg/s respectively in experimental group;(34.48?4.68) mm Hg,(19.27?4.63) mm Hg and (935?196) mm Hg/s respectively in control group;(32.14?4.95) mm Hg, (16.99?4.85) mm Hg and (825?302) mm Hg/s respectively in solution group.The hemodynamic indexes in experimental group were superior to those in control group and solution group (P

Objective To investigate the effects of Jiawei Wuzi Yanzong Pill and its components on central cholinergic system in the model mice with learning and memory disorders induced by scopolamine. Methods Mice were treated by Jiawei Wuzi Yanzong Pill and its components for 15 days and induced by ip scopolamine. After the experiment in Morris water maze, the content of acetylcholine and the activity of acetylcholinesterase from brains of mice were determined by colorimetry. Results Significant differences were observed between model group and naked group that the content of acetylcholine and the activity of acetylcholinesterase from brains of model mice were decreased (P 0.05). Conclusion Jiawei Wuzi Yanzong Pill and its components could improve the function of central cholinergic system by increasing the content of acetylcholine and the activity of acetylcholinesterase and possesses a certain preventive effects on mice with learning and memory disorders induced by scopolamine.