BACKGROUND:Chinese herb, bushen yizhi formula protects at certain extent learning and memory in rat model of Alzheimer disease. The drug serum in this formula can alleviate neurotoxic reaction of nerve tumor cell NG 108-15 to beta-amyloid protein. In order to understand further the mechanism and compatibility of the formula, it is necessary to carry on the study on the broken formulas.OBJECTIVE:To study the effect of drug serum in subgroups of broken bushen yizhi formulas on growth and differentiation of cell model of Alzheimer disease and probe into the compatibility rule of bushen yizhi formula in view of serum pharmacology.DESIGN: Randomized controlled experiment.SETTING: DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine.PARTICIPANTS:The experiment was performed in DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine from January to August 2003, in which, 40 healthy male SD rats of 3 months old were employed and NG108-15 cell line was frozen-preserved.into the control, original formula group (No. 1 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), renshen (Panax ginseng C.A.Mey.), heshouwu (Polygonum multiflorum Thunb.), danpi (Paeonia Suffruticisa Andr.) and bingpian (Borneolum)], kidney replenishment group (No.2 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), etc.], group for benefiting qi and nourishing blood (No.3 group)[renshen (Panaz ginseng C.A.Mey), zhishouwu (Polygonum multiflorum Thunb.), etc.] and group with bingpian (Borneolum) removed (No.4 group)[Panax ginseng C.A.Mey.], heshouwu (Polygonum multiflorum Thunb.) and danpi (Paeonia Suffruticisa Andr.)], 8 rats in each group. The concentrated Chinese herbal solutions of every group were applied at 10 μL/g (equal to 6 g/kg of raw herbs) for gastric infusion successively,continuously for 1 month. In the control, the physical saline solution of equal dosage was used for infusion. Two hours after the last gastric infusion in rats of each group,the blood was collected from heart after anesthesia and the serum was sepaNG108-15 cell cultured in vitro was divided into 6 groups. In the control and model group, normal rat serum was contained in proliferated culture solution. In the rest 4 groups, the drug serum of No. 1 group and 3 sub-groups was contained.Simultaneously, beta-amyloid protein 25-35 in each hole was prepared to the terminal concentration 5 μmol/L (except in the control) and the culture went on for 48 hours.MAIN OUTCOME MEASURES:MTT method was used to determine proliferated number and survival rate of cells. Simultaneously, the ratio of neurite cells to total cell count and average length of neurit were determined.icantly than the control (0.520±0.022, 0.665±0.037, P ＜ 0.01), and that in every drug serum group was higher than model group, of which, the result vival rate of differentiated cells: That in model group was lower significantly than the control (58.4%, 100%) and that in every drug serum group was higher than model group, of which, the result in No.4 group was the most tal cell count: That in model group was lower significantly than the control [(42.95±11.42)%, (58.75±12.84)%, P ＜ 0.01] and that in every drug serum group was higher than model group, of which, the result in No.4 group was rite: That in model group was shorter significantly than the control [(356.0 ±109.0), (493.8±133.0) μm, P ＜ 0.01] and that in every drug serum group was longer than model group, of which, the result in No.4 group was the most significant [(486.8±79.2) μm, P ＜ 0.01].CONCLUSION: The drug serum in all of bushen yizhi formula and every subgroup inhibits at certain extent the injury of beta-amyloid protein 25-35 to NG108-15 cell, but the results of each group are various. The protection of drug serum to the cell in every group is in the sequence from strong to weak as group with bingpian removed ＞ original formula group ＞ kidney replenishment group ＞ group for benefiting qi and nourishing blood. It is to expect a further study on the efficacy of group with bingpian removed.

Object To observe the improving effects of BUSHEN YIZHI DECOCTION (BSYZD) on interspace explore learning and memory in rat model with Alzheimer's disease. Methods Eighty 15-month Wistar rats were induced by ip D -galactose for four weeks and injection of basal nucleus of Meynert with ibotenic acid (IBO) to make AD model, then randomly divided into AD model group, Hup-A treated group, BSYZD (high dose, 12 g/kg?d) treated group and BSYZD (low dose, 6 g/kg?d) treated group, and also normal aged and young groups. After treating for four weeks, Morris water maze was used to assess the improvement of rat interspace explore learning and memory. Results In the interspace explore experiment, the significant differences were observed between the model, Hup-A treated groups and normal aged, young groups (P0.05). Conclusion BSYZD possesses a certain preventive effects on interspace explore learning and memory in AD rat model.

Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.

0.05). The peak of NAA was significantly different between high grade glioma and metastatic carcinoma(P0.05). The peak of AA was characteristic of brain abscess. The ratio of Cho/Cr0 in brain abscess was significantly lower than those in high grade glioma and metastatic carcinoma(P

OBJECTIVE:To evaluate the management of the free anti-TB drugs in He'nan province,and to analyze ex-periences and find out the gap.METHODS:Based on the requirements of the Chinese TB Control Program-Free anti-TB drug management manual,a questionnaire comprised of 17 items was developed and 20 TB drug storerooms at city or county level were randomly sampled for on the spot investigation.RESULTS:Of the total drug storerooms investigated,75% had yearly drug demand plan,but only 35% was up to the standard in inventory control,95% had no expired drug,90% had inventory/supply vouchers and detailed inventory records,only 25% achieved conformity between records and physical counts.The condition of drug storerooms and the storage of drugs were unable to meet the requirement.CONCLUSION:Tuberculosis Control Agency at different levels haven't paid due attention to the management of free anti-TB drugs.The personnel in this agency should raise their drug management responsibility from aspects of saving public belongings and ensuring patients' medication quality.

Objective To identify gene expression profile in periphend blood mononuclear cell (PBMC)of patients with primary Sjogren's syndrome(pSS)complicated with hematology involvement and compare it to healthy volunteers.Methods Three Sjogren's syndrome(SS)patients with leucopenia and 3healthy volunteers were included.The cRNA probes prepared for total RNA were hybridized with three identical gene chips.The difference of gene expression of each patient and volunteers were compared.Results Significant difference in the expression of 82 genes could be detected between patients and volunteers.Among these,45 were upregulated,and 37 were downregulated.Statistical difierence was calculated between patients and volunteers especially in the following two pathways:the complement and coagulation pathways(including C2,PROS1,F2R and SEPPING1)and the cytokine-cytokine receptor interaction pathway(including FASLG,MPL,CCL20 and CXCL2)(P<0.01).Conclusion This study has identified distinct gene expression profiles in PBMC from patients with pSS patients with hematology involvement and healthy volunteers.This provides new knowledge about groups of genes that are upregulated during disease.It may form an excellent platform for future functional studies.

ObjectiveTo observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.MethodsThirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.ConclusionThe effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

Objective To study whether the primary ovarian cancer cells containing cancer stem cells and its characterization in serum-free culture condition.Methods The primary cancer cells were isolated from one stage Ⅲ, grade 2 serous adenocareinoma tissue.Cells were cultured in serum-free culture system supplemented with epidermal growth factor,basic fibroblast growth factor,leukemia inhibitory factor and insulin.or standard serum-containing system.Methyl thiazolyl tetrazolium assay,quantitative PCR analysis,flow-cytometric analysis and xenograft experiments in vivo were performed.Results The primary cancer cells could maintain and forill cell sphere in serum-free culture system.These cells had the properties of self-renewal,overexpression of stem cell marker genes Nanog,Oct-4,Sox-2,nestin,ABCG2,CD_(133) and CD_(117) By contrast with the difierentiated cells under standard serum-containing culture conditions.these sphere-forming cells were more resistant to cisplatin and paclitaxel after treated 48 and 72 hours(61% vs.31%,73% vs.29%,P<0.05).With Hoechst 33342 exclusion assay,only 21.83%of sphere-forming cells were positive with the dye,compared with 83.04% positive cells in differentiated cells (P<0.01).Only 500 sphere-forming cells resulted subcutaneous xenograft tumors.All of these xenografts were categorized as serous adenocareinomas, overexpression of CA_(125) and cytokeratin-7 which were original tumor phenotype of ovarian cancer. Conclusion The sphere-forming cells isolated from primary ovarian cancer tissues have the characterization of cancer stem cell and may be a more reliable model system for understanding the biology of primary human tumors.

Objective To investigate signal transduction and cytokine gene expression pattern in peripheral blood mononuclear cell (PBMC) between primary Sjogren syndrome patients and healthy controls. Methods The PBMC of 3 patients with Sjogren syndrome and 3 healthy volunteers with consistent age were collected. The total RNA extracted from the PBMC was carried the reverse transcription and in vitro transcription (IVT), then labeled with Cy5/Cy3 and hybridized on the gene chips. After scanning and data extraction with LuxScan 3.0, differentially expressed genes were analyzed with SAM method. The online tools of molecule analysis system (MAS) was used for biological knowledge mining. The difference of signal transduction and cytokine gene expression of each patient and control were compared. Results Statistical difference was calculated between the patient and control group in the following four pathway: the chemokine-cytokine receptor interaction pathway[(FASLG(Fas ligand), CCL5, CCR3, IL-4R, CXCL10 (CXC chemokine ligand 10), TNFRSF19L (tumor necrosis factor supeffamily 19 ligand), CX3CR1 (CX3C chemokine receptor 1)], adhension molecule pathway [interleukin adhesion molecules-1 (ICAM-1)], Jak-STAT signal pathway [STAT4 (signal transducer and activation of transcription 4), CISH (chromogenic in situ hybridization), IL-4R], Toll-like receptor signal pathway(CCL5, CXCL10). Among these, 4 genes were up-regulated and 6 genes were down-regulated. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjogren syndrome.