OBJECTIVE: To study the monoterpenoid indole alkaloids in Gardneria multiflora Makino. METHODS: The dried plants of G. multiflora Makino were extracted with 95% ethanol. After removal of solvent, the residue was diluted with water, then extracted with petroleum ether and chloroform to obtain crude alkaloids. Column chromatograghy on silica gel, Rp-18, and Sephadex LH-20 were applied for the isolation and purification of crude alkaloids. The structure was elucidated based on detailed spectroscopic evidence. RESULTS: One monoterpenoid indole alkaloid was isolated from this plant. CONCLUSION: Compound gardmultimine A (1) is a new compound.

Objective To investigate the effects of erythropoietin(EPO)on the function of glomerular endothelial cells in rats with chronic renal failure(CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats.Experimental rats were randomly divided into four groups:sham operation group (control group),CRF group,CRF rats treated with 30 U/kg EPO(low-dosage group)and with 50 U/kg EPO (high-dosage group).CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed.Serum creatinine(Scr),blood urea nitrogen fBUN),urine protein,haematoglobin (Hb) and blood pressure were measured.The renal morphologie changes were evaluated on periodic acid-schiff (PAS) stained sections.The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method.The mRNA of endothelin 1(ET-1),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus,and the expressions of eNOS and VEGF roRNA in renal tissue were higher in EPO treatment group than those in CRF model group(all P<0,05).The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group.In addition,the Scr,BUN,urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05).Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05).Reanl pathological injury wss improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure,maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

Objective To investigate the effects of erythropoietin (EPO) on the number and function of peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). Methods The model of chronic renal failure was established by a two-stage 5/6nephrectomy procedure in rats. Experimental rats were randomly divided into four groups (n =7,respectively): sham operation group, CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats were given EPO by hypodermic injection for 6 weeks, then EPCs were isolated by density gradient centrifugation from peripheral blood mononuclear cells. The ability of cell proliferation, adhesion and vasculogenesis in vitro was further observed. Results Compared to sham operation group, the ability of cell proliferation,adhesion and vasculogenesis in vitro in CRF rats was remarkably decreased (P＜0.05, respectively).Such ability was promoted significantly in dose-dependent manner by EPO treatment (P＜0.05,respectively). Conclusion EPO can improve the number and ability of endothelial progenitor cells from rats with chronic renal failure.

Objective To evaluate the effects of tadalafil on chronic allograft vasculopathy. Method The abdominal aorta transplantation was performed on Male Lewis or Brown-Norwai rats as donors and male Male Lewis rats as recipients. The recipients were divided into 3 groups: Group A (Lewis-Lewis)with no treatment ;Group B (BN-Lewis) with no treatment; Group C (BN-Lewis) received tadalafil treatment (0.5 mg/kg per day). The rats were sacrificed at 8 weeks post treatment. The grafted aortas were used for histology and Western blot assay. Plasma cGMP level was detected by ELISA assay. Results The aortas intimal in group C was significantly thinner than that in group B. PKG-Ⅰprotein expression in group C was significantly higher than that in group B. Expression of RhoA in group C was lower than that in group B. Conclusion Tadalafil has positive effect on chronic allograft vasculopathy.

Child, Preschool ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Diseases ; drug therapy ; Humans ; Infant ; Male ; Phytotherapy ; Plant Extracts ; therapeutic use

Risk Factors ; Tars ; toxicity ; Tobacco Products ; standards ; toxicity ; Tobacco Use Disorder ; etiology ; prevention & control

OBJECTIVETo explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method.

METHODSThe retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05).

CONCLUSIONSAGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

Animals ; Cells, Cultured ; Diabetic Retinopathy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ependymoglial Cells ; Glucose ; L-Lactate Dehydrogenase ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A

Background and purpose:TrkB,a neurotrophic tyrosine kinase receptor,serves as a potent and specific suppressor of anoikis of non-malignant epithelial cells,and induces high invasion capacity in these cells.TrkB over-expression has been associated with chemotherapy resistance and poor survival in neuroblastoma and some other highly aggressive cancers.However,the relationship of the expression of TrkB to anoikis resistance in ovarian cancer cells has rarely been reported in literature.This study investigated the expression and significance of anoikis- suppressor TrkB in OVCAR-3 ovarian cancer cells.Methods:The expression of TrkB and its ligand BDNF was evaluated in OVCAR-3 ovarian cancer cells under different culture conditions by RT-PCR and Western blot.Results: TrkB mRNA was overexpressed in multicellular spheroids(cell-spheroids,anchorage-independent culture,AIC)as compared to that in OVCAR-3 cells(adhesive-cells,adhesive culture,AC),(35.3?0.7)% versus(23.5?0.5)%;but BDNF mRNA expression was the opposite to the above situation,(41.4?0.6)% versus(32.24?0.7)%(P

BACKGROUND: Schwann cells play an important role in axonal growth and myelin sheath formation of the peripheral nerve. Whether Schwann cells play the same role in the spinal cord had attracted considerable attention. Microencapsulation technology as an effective immune isolation technique can effectively keep Schwann cell activity to play the repair effect of Schwann cell in the spinal cord.OBJECTIVE: To observe the changes of myelin sheath in the injured transection of rats after transplantation of the alginic acid microencapsulated Schwann cells.DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Basic Medical School of Nanchang University from March 2005 to February 2008.MATERIALS: Sciatic nerve trunk was obtained from adult rabbits to harvest Schwann cells in vitro using repeatedly differential velocity adherent technique, and to prepare Schwann cell suspension and microencapsulated Schwann cell suspension.METHODS: A total of 146 adult Sprague Dawley rats were used to establish models of right hemi-transection damage at T_(10) level and randomly assigned to four groups: simple injury group (n=44), cell transplantation group (n=44), microencapsulated cell transplantation group (n=44) and normal control group (n=14). At 1, 3, 7,14 and 28 days following surgery, 8 rats were selected from each group at each time point (2 from the normal control group) for perfusion and fixation. Spinal cord tissue was collected to make paraffin section, and then subjected to hematoxylin-eosin staining and Loyez myelin staining. In addition, 2 rats were selected from each group at 2 and 8 weeks. The spinal cord tissue was fixed, embedded in Epon816, stained using uranyl acetate and aluminum citrate, and then observed using an electron microscope.MAIN OUTCOME MEASURES: Neuron number and survival were observed surrounding the damaged region. Structural changes in the myelin sheath from spinal cord white substance at the damage site were measured.RESULTS: At 1 and 3 days following spinal cord injury, spinal neurons were degenerated and necrotic at damaged site, with reduced number of myelin sheath, loose structure, but above-mentioned was rare in the cell transplantation and microencapsulated cell transplantation groups. At 7 days, the reduced number of myelin sheath, with damaged structure was seen. The microencapsulated cell transplantation group was light. At 14 days, number of neurons was increased, with increased cell body, especially in the microencapsulated cell transplantation group. At 28 days, neurons gradually recovered, myelin sheath was gradually complete, with increased number in the microencapsulated cell transplantation group. There were significant differences compared with the simple injury and cell transplantation groups (P < 0.01). At 8 weeks, abundant myelin sheath was repaired, with new myelin sheath in the microencapsulated cell transplantation group.CONCLUSION: Microcapsule has immune isolation effects. Microencapsulated rabbit Schwann cells can promote the repair of rat spinal cord neurons and axonal myelinization.

Background Aniridia is a rare congenital hereditary eye disease.Studies determined that PAX6 gene mutation is closely associated with congenital aniridia,but the mutation locus are varied.Objective This study was to identify virulence mutation locus of PAX6 gene of a Chinese family pedigree with autosomal dominant aniridia.Methods A Chinese family affected with autosomal dominant aniridia was collected and examined in Affiliated First Hospital of Zhengzhou University in August 2014.Periphery blood of 10 ml was collected from all the families and 100 unrelated health controls.The genomic DNA was extracted by standardized phenol-chloroform method,and all exons and splicing junctions of PAX6 were amplified by PCR.Real-time fluorescence quantitative PCR was performed to examine the relative expression of PAX6 mRNA in patients and normal phenotype families and heahh controls.This study protocol was approved by Ethic Committee of Affiliated First Hospital of Zhengzhou University and complied with Helsinki Declaration.Written informed consent was obtained from subjects or custodian before any medical examination.Results This Chinese family inclued 3 generations and 9 members,with a classic autosomal dominant inheritance mode.Five patients were found,showing the absence of iris and cataract in 3 adult patients and only absence of the iris in 2 children,and other 4 members showed the normal phenotype.A novel heterozygous PAX6 deletion mutation c.796 del G (p.A266 fs) (GenBank ID:KP255960) in exon 10 was exclusively found in all affected individuals but not in any of the unaffected families or unrelated health controls.PAX6 mRNA level in lymphocytes was about 50％ lower in aniridia patients than in unaffected family members,indicating that this mutation caused nonsense-mediated mRNA decay.Conclusions A novel deletion mutation in PAX6 gene results in an abnormal PAX6 COOH-terminal extension in the Chinese aniridia family.This finding expands the mutation spectrum of PAX6 gene.