The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment,and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied.The results showed that B5 medium supplemented with 0.5mg/L NAA,0.6mg/L 6-BA and 30g/L sucrose,at initial pH 5.0～5.5,20g(FW)/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E.ulmoides suspension cells growth and flavonoids synthesis.The results of metabolic kinetics analysis for E.ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth,sucrose consumption and flavonoids production during the process.The maximum specific growth rate(?m),the actual growth yield based on sucrose(YG) and maintenance coefficient(m) were 0.417/d,0.619g/g and 0.0206g/(g?d-1) respectively.All these outcomes could give a basis for establishing the suspension cell culture of E.ulmoides and production of the natural active components in large-scale.

Higenamine (HG) is a potent cardioactive benzylisoquinoline alkaloid isolated from Aconiti tuber which has long been used as a cardiotonic in traditional Chinese medicine. HG exerts various effects on the cardio-circulatory system inotropic and chronotropic in isolated rat atria. It also relaxes isolated rat aorta. It inhibits epinephrine, ADP or collagen-induced platelet aggregation in platelet rich plasma. HG inhibits LPS-induced nitrate accumulation and the expression of iNOS mRNA in RAW 264.7 cells. HG lowers blood pressure in rats and increases the recovery rates in acute thrombosis model of mice, and lower the weight of thrombus formed in the arterio-venous shunt model of rats. Higenamine also has ameliorative effects in the LPS-induced DIC model.
Aconitum ; chemistry ; Alkaloids ; isolation & purification ; pharmacology ; therapeutic use ; Animals ; Blood Pressure ; drug effects ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Disseminated Intravascular Coagulation ; drug therapy ; metabolism ; Fibrinolytic Agents ; pharmacology ; therapeutic use ; Humans ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type II ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; RNA, Messenger ; biosynthesis ; genetics ; Tetrahydroisoquinolines ; isolation & purification ; pharmacology ; therapeutic use

OBJECTIVETo investigate the effects of Tianzhi Keli (TZ) on acetylcholine (ACh) and catecholamine levels in striatum of rats with neuromitochondrial impairment, and try to find out the neuroprotective mechanism of TZ.

METHODThe microdialysis and high performance liquid chromatography (HPLC)-post column Immobilized enzyme reactor (IMER)-electrochemical detection (ED) were used to establish a model of mitochondrial energy metabolism impairment which induced by perfusion with sodium azide (NaN3), and measure continuously the effects of TZ on extracellular ACh, choline (Ch) and catecholamine of model rats.

RESULTAfter perfusion with NaN3, ACh, noradrenalin (NE), adrenaline (E), dopamine (DA), 3,4-Dihydroxyphenyl-aletic (DOPAC), and homovanillic acid (HVA) levels were decreased obviously (P < 0.05-0.01), while Ch level was increased distinctly (P < 0.01). Transmitters levels were recovered individually after stop the perfusion with NaN3. TZ can postpone the decrease of ACh and advance the recover of Ch. The effect of TZ coupled with duxil on increasing ACh level is more obviously than effect of TZ or duxil. TZ is also showing a tendency to postpone the decrease of catecholamine and advance its recovery. TZ coupled with duxil can advance the recovery of DOPAC and adjust the metabolic abnormity positively.

CONCLUSIONTZ has effect on protecting impairment of choline neurosystem, which induced by damage of mitochondrion and abnormity of energy metabolism; coupled with duxil have synergistic action. TZ also has tendency to protect the impairment of epinephrine and dopamine neurosystem.

3,4-Dihydroxyphenylacetic Acid ; metabolism ; Acetylcholine ; metabolism ; Animals ; Catecholamines ; metabolism ; Corpus Striatum ; metabolism ; Dopamine ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Extracellular Space ; metabolism ; Gastrodia ; chemistry ; Male ; Microdialysis ; Mitochondrial Diseases ; chemically induced ; metabolism ; Norepinephrine ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sodium Azide ; Uncaria ; chemistry

OBJECTIVETo investigate the effect of Tianma Gouteng recipe (TGR) on interfering left ventricular (LV) and aortic hypertrophy and tissue angiotensin II (Ang II) in rats with renovascular hypertension.

METHODThe animal model of renovascular hypertension was used in this experiment. Hypertensive rats were randomly allocated into model group, Enalapril group and TGR group, and the drugs were used for 6 weeks continuously. During this period, the blood pressure of rats was measured every two weeks. After rats were sacrificed, the wet weight, tissue Ang II level of LV and aorta, and the cardiac index were measured.

RESULTOne week after renovascular stenosis, the systolic blood pressure (SPS) of model group was increased by 37.4 mmHg, and 7 weeks after stenosis, the LV and aortic hypertrophy was obvious increased, meanwhile, tissue Ang II of LV and aorta was raised markedly (P < 0.01). Contrasting with the model group, blood pressure was reduced and the morphological index was improved in Enalapril group respectively (P < 0.05 or P < 0.01); the wet weight of LV and aorta were reduced, the morphological index was improved, the rise of Ang II in tissue was suppressed, in TGR group significantly.

CONCLUSIONTGR can attenuate myocardial and aorta hypertrophy induced by renovascular hypertension, and suppress the rise of Ang II in tissue significantly. This suggests that TGR has the effects on interfering LV and aortic hypertrophy by an independent-antihypertensive way.

Angiotensin II ; metabolism ; Animals ; Antihypertensive Agents ; pharmacology ; Aorta ; metabolism ; pathology ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Enalapril ; pharmacology ; Gastrodia ; chemistry ; Hypertension, Renovascular ; complications ; physiopathology ; Hypertrophy, Left Ventricular ; etiology ; metabolism ; pathology ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Uncaria ; chemistry

The effects of Gastrodia elata on preventing decrepitude and advancing memory are closely associated with its neuroprotective activity. Previous researches proved that G. elata, its active components and preparations played a neuroprotective role by affecting the excitotoxicity, nitric monoxide (NO) system, neuroglia, biomembrane, oxidative neurotoxicity, apoptosis et al. Recent researches also suggest that reducing energy metabolism impairment, anti-inflammatory and immune modulating function may be new research targets of neuroprotective mechanism of G. elata.
Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Excitatory Amino Acids ; antagonists & inhibitors ; Gastrodia ; chemistry ; Humans ; Neuroprotective Agents ; pharmacology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Plants, Medicinal ; chemistry

Antiviral Agents ; isolation & purification ; pharmacology ; Croton ; chemistry ; Egypt ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Glucosides ; chemistry ; isolation & purification ; pharmacology ; HIV-1 ; drug effects ; Molecular Structure ; Phorbol Esters ; chemistry ; isolation & purification ; pharmacology ; Phyllanthus emblica ; chemistry ; Plant Extracts ; pharmacology ; Plants, Medicinal ; chemistry ; Virus Replication ; drug effects

BACKGROUNDIt is generally accepted that gliomas are the most common primary brain tumors with poor prognosis. We aimed to explore the relationship of the immunity of the central nervous system and the genesis and development of glioma.

METHODSG422 glioma was implanted in the brain of BALB/c mice (immuno-competent mice), nude mice (T cell related immuno-deficient) and complement C3 knock-out mice (complement C3 related immunodeficient). The survival time of the host, growth and histopathology of the tumor, and concentrations of tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ) in tumor tissues were assessed.

RESULTSTumor spheres were formed in all mice after injection, and glial fibrillary acidic protein (GFAP) positive staining of the cells declared their glioma origin. The longest median survival time of (44.3 ± 6.0) days was found in BALB/c mice, followed by (24.8 ± 5.2) days in nude mice and the shortest (18.6 ± 5.8) days in complement C3 knock-out mice. Accordingly, the growth of the tumor was fastest in complement C3 knock-out mice, followed by the nude mice and slowest in the BALB/c mice. Although the proportions of infiltrating CD68(+) lymphocytes in tumor tissues showed no significant difference (P > 0.05), TNF-α level in the nude and C3 knock-out mice, (28.11 ± 4.86) µmol/L and (22.87 ± 6.36) µmol/L respectively, were significantly lower (P < 0.01) than that in the BALB/c mice, which was (230.21 ± 39.17) µmol/L. The INF-γ level was highest in the BALB/c mice ((180.76 ± 29.19) µmol/L), followed by the nude mice ((113.46 ± 23.76) µmol/L) and then the C3 knock-out mice ((16.84 ± 4.45) µmol/L).

CONCLUSIONSThe G422 glioma implanted in the brains of mice with different immune ability would be a useful model for studying the relationship of the immune system and tumor in the central nervous system. Furthermore, the T cells and complement C3 compartments of the immune response may affect the growth of implanted tumors and inflammatory factors such as TNF-α and INF-γ.

Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; Complement C3 ; genetics ; metabolism ; Glioma ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Mice, Nude ; Tumor Necrosis Factor-alpha ; metabolism

The Bacillus subtilis Sac B gene with the vacuolar targeting signal sequence driven by 35S promotor was transferred into Lespedeza thunbergii by Agrobacterium mediated method. Total 62 Kan-resistant plants were obtained, of which 5 plants were proved to be transgenic plants. The transgenic plants were characterized by PCR amplification, PCR-Southern hybridization and RT-PCR. The physiological assay results showed that the transgenic plants were more tolerant to stress than the controls under the condition of 200mmol/L NaCl and 5% PEG, respectively, and that the content of soluble sugar in trnsgenic plants was significantly higher than that of controls in the period of tests (5-15 days) under salt and PEG stress.
Bacillus subtilis ; enzymology ; genetics ; Carbohydrate Metabolism ; Carbohydrates ; chemistry ; Hexosyltransferases ; genetics ; Lespedeza ; drug effects ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium Chloride ; pharmacology ; Solubility ; Stress, Physiological ; drug effects ; Transformation, Genetic ; Transgenes ; genetics

OBJECTIVETo discuss the treatment and mechanisim of acetabulum fracture combined with ipsilateral lower extremity fracture.

METHODSFourteen cases(9 males and 5 females) of acetabulum fracture were involved in the study with the mean age of 35 years (range, from 18 to 65 years). According to Letournel classification, 11 cases were the fracture of posterior wall and column, 3 cases were only the fracture of posterior column. All of them were treated with titanium plate internal fixation. Of three cases with ipsilateral femoral intertrochanteric fracture, one was treated with external fixiation device, other two were treated with DHS fixation; Three cases with ipsilateral femoral neck fracture were treated with compressive cannulated screw fixation; Among six cases with ipsilateral femoral shaft fracture,one was fixed with steel plate,the rest underwent intramedullary nailing fixation; Two cases with ipsilateral tibial plateau fracture were treated with anatomic plate internal fixation.

RESULTSAll patients except the died old patient were followed up from 18 months to 5 years, with mean of 30 months. According to standard of American institute of orthopaedics, 9 cases obtained excellent results, 3 good and 2 poor and the excellent and good rate was 92.3%.

CONCLUSIONMost cases of acetabulum fracture combined with ipsilateral lower extremity fracture are due to great violence, their mechanisms are complex, and easily lead to missed diagnosis, therefore the early correct diagnosis, and reasonable internal fixation should be considered.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged

OBJECTIVETo study the effect of cyclooxygenase-2 (COX-2) on lymphangiogenesis in breast cancer.

METHODSBy the means of immunohistochemistry, COX-2, vascular endothelial growth factor-C (VEGF-C) and D2-40 were examined in the tissue samples of primary tumors from 94 patients underwent surgical resections for breast cancer from November 1998 to March 2002. Eighty-three patients were followed-up. The expressions of VEGF-C mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in MDA-MB-231 cell lines by the treatment of selective COX-2 inhibitor Nimesulide at different doses. The expressions of VEGF-C protein were evaluated in MDA-MB-231 cells treated by PGE2 (1 microg/ml) and Trastuzumab (1 microg/ml), respectively.

RESULTSCOX-2 over-expression was observed in 46.8% of surgical specimens (44/94), while VEGF-C overexpression occurred in 51.1% of tumor samples (48/94). COX-2 was strongly correlated with VEGF-C expression (P < 0.01), micro-lymphatic vessels (P = 0.032) and metastatic lymph nodes (P = 0. 035). Patients with COX-2 positive tumors had a significant shorter survival time than those with negative tumors did, including disease-free survival (P = 0.010) and overall survival (P = 0.040). Nimesulide could down-regulate the expressions of VEGF-C mRNA and protein in a does-dependent manner, while PGE2 could up-regulate the expressions. The expression of VEGF-C protein up-regulated by PGE2 treatment was decreased by Trastuzumab.

CONCLUSIONSCOX-2 over-expression can up-regulate the expression of VEGF-C. VEGF-C might promote lymph node metastasis by a lymph-angiogenic pathway, then affect the prognosis of the patients with breast cancer.

Adolescent ; Adult ; Aged ; Breast Neoplasms ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; physiology ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor C ; genetics ; metabolism