According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
Cloning, Molecular ; Computational Biology ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase ; genetics ; Panax notoginseng ; genetics ; Real-Time Polymerase Chain Reaction

Brucellosis ; complications ; Cytomegalovirus Infections ; complications ; Humans

Objective To explore the effect of Okam on airway inflammation in asthmatic mouse.Methods Thirty-two SPF grade Kunming Strain mice were randomly divided into positive control group,glucocorticoid inhalation group,Okam group and negative control group with 8 mice in each group.The mice were sensitized and repeatedly challenged with ovalbumin(OVA) to establish the models of chronic asthma.The glucocorticoid group were given Budesonide(200 ?g) and saline everyday by inhalation,the Okam group were given 50 mg/kg Okam by gavage,and the positive group had saline at the same time,the negative control group received saline at all stages.The inflammation of the lung tissue were scored underwent HE staining.Bronchoalveolar lavage fluid(BALF) cell count and differential were studied,and interferon-?(IFN-?),interleukin-4(IL-4) in BALF were determined by enzyme-linked immunosorbent assay(ELISA).Results There were no inflammatory cell infiltrate of bronchiole in the negative control group.Inflammatory infiltration of lung tissue were obvious in the positive control group.Inflammatory infiltration of lung tissue lightened obviously in the Budesonide and Okam groups.The total cell number,Eosinophils(EOS) and IL-4 level in BALF,and the score of the lung tissue in Okam group were all markedly lower than those in positive control group(t=5.942,7.089,7.078 Pa0.05),IFN-? lower(t=4.275 P

AIM: To investigate the expression of NKG2A,NKG2D and their ligands in pregnancy uterine micro-environment and to probe the function of NKG2A and NKG2D imbalance expression during the immunotolerance at the fetal-maternal boundary.METHODS: Decidual lymphocytes and peripheral lymphocytes were obtained from 30 women during 6-9 weeks of pregnancy who were undergoing selective termination.FACS technology was used to detect NK cells number and NKG2A,NKG2D expression.RT-PCR was used to investigate HLA-E and MICA mRNA in trophoblast tissue.RESULTS: Natural killer cells predominate,accounting for 70% of pregnancy endometrial lymphocytes.FACS results indicated that NKG2A was significantly increased in decidual NK cells as compared with that in peripheral NK cells,accounting for 97.86%?1.75% and 33.35%?10.92%.The difference between them in NKG2A expression was significant(P

OBJECTIVETo develop a urine pretreatment method of Solid Phase Extraction (SPE) for the quantitative determination of a number of aristolochic acids (AAs) and aristololactams (ALs) in rat urine.

METHODThe HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other sixteen AAs and ALs was chosen as evaluating index to study the extract results of five Solid Phase Extraction columns (Agilent C18/100 mg, Alltech HG18/100 mg, Alltech C18/100 mg, Alltech C18/300 mg and Agilent Phenyl/200 mg) comparatively. The influences of two washing solvents (water and 1% acetic acid-0.02% triethylamine solution) and seven eluting solvents (ether, acetone, chloroform, ethyl acetate, dichloromethane, methanol and acetonitrile) on extract results of AAs and ALs are comparatively studied with the extracting recoveries of AA-I , AA-II, AL-I and AL-II as indicators. The HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other seven AAs and ALs with good separation being targets, several factors which affect extracting efficiency of analytes, including activating volume, cleansing volume, washing volume and eluting volume, are optimized by orthogonal design experiments with four factors at three levels.

RESULTThe established method of SPE is as follows: Agilent Phenyl SPE column of 200 mg, activating with 1.0 mL methanol, cleansing with 1 mL water, adding 1.0 mL rat urine sample, washing with 0.8 mL 1% acetic acid 0.02% triethylamine solution, and eluting with 3.0 mL methanol.

CONCLUSIONThe established method of SPE is efficient, selective, simple and fast, and can be used as urine pretreatment method to analyze a variety of aristolochic acids and aristololactams in rat urine.

Administration, Oral ; Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; urine ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Solid Phase Extraction ; methods

OBJECTIVETo investigate the effect of hypoxia inducible factor-1 alpha (HIF-1 alpha) on vascular endothelial growth factor (VEGF) expression in Tca8113 cells under hypoxia.

METHODSThe expression of the mRNA of HIF-1 alpha and VEGF in Tca8113 cells was examined by RT-PCR technique at different culture times (1/2 h, 1 h, 3 h, 6 h, 12 h, 24 h) under normoxic and hypoxic conditions.

RESULTSThe expression of HIF-1 alpha under hypoxia showed the trend of increasing first and then decreasing, and was higher than that of the control (normoxic group) at 6h and 12 h (P < 0.05). The expression of VEGF under hypoxia was higher than that of the control group at 1/2 h, 1 h, 3 h, 12 h, 24 h (P < 0.05). The expression of hypoxia-induced VEGF mRNA increased with the increased expression of HIF-1 alpha mRNA in the cell lines tested at the initial stage of hypoxia. But no statistical significant association was observed between HIF-1 alpha and VEGF expression within 24 h under hypoxia (rs = 0.5750, P > .005).

CONCLUSIONSThe increased expression of VEGF in Tca8113 cells might be mediated by multiple factors, including HIF-1 alpha.

Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Hypoxia ; genetics ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; RNA, Messenger ; genetics ; Tongue Neoplasms ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the difference of soluble P-selectin levels in different subtype of coronary heart disease and the relationship between soluble P-selectin levels with the severity of coronary artery lesions.

METHODSEnzyme linked immuoserbent assay (ELISA) was used to measure the plasma soluble P-selectin levels in 69 patients with angiocardiography documented coronary heart disease and 19 normal coronary arteries persons without angiocardiography detectable coronary artery disease (control group). The coronary artery lesions score was recorded according to single, double and triple-vessel lesions while the American College of Cardiology and the American Heart Association proposed type A, B, C lesion and Gensini scoring system. The relationships between plasma soluble P-selectin levels and the coronary artery score (the severity of coronary heart disease) were assessed.

RESULTS(1) The level of plasma soluble P-selectin was obviously higher in the coronary heart disease group than in the control group (180.6 +/- 60.5 ng/L vs. 145.3 +/- 21.7 ng/L, P<0.05). (2) The level of plasma soluble P-selectin was significantly higher in the acute coronary syndrome group (191.4 +/- 63.7 ng/L) than in the control group (145.3 +/- 21.7 ng/L, P< 0.01) and in the stable angina pectoris group (141.3 +/- 17.9 ng/L, P<0.01). (3) The level of plasma soluble P-selectin was high in multi-vessel coronary artery lesions group than in single-vessel group (190.1 +/- 64.2 ng/L vs. 157.2 +/- 43.4 ng/L, P < 0.05). The level of plasma soluble P-selectin was positively correlated with the Gensini score (r = 0.391, P = 0.001); the numbers of vessels lesions (rs = 0.349, P = 0.003); Type A, B and C lesions (rs = 0.358, P = 0.002).

CONCLUSIONThe positive correlation between the level of soluble P-selectin and the coronary artery score may indicate that soluble P-selectin levels might reflect the severity of coronary heart disease. The elevated soluble P-selectin level in acute coronary syndrome suggested the possible relation of P-selectin to the pathogenesis of acute coronary syndrome, which may save as a potential marker of plaque unstability.

Case-Control Studies ; Coronary Disease ; blood ; physiopathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Middle Aged ; P-Selectin ; blood ; chemistry ; Solubility

OBJECTIVETo study pharmacodynamic characteristics by oral administration aristolochic acid I (AA-I) in rats.

METHODAfter one-time oral administration of Aristolochiae manshuriensis decoction 10 g x kg(-1) and 125I labeled AA-I (containing AA-I 37.2 microg x mL(-1)), whole blood concentration of 125I-AA-I and the binding rate of serum albumin were detected in 69 normal wistar male rats. Metabolic dynamic parameters were calculated by program 3P87 with a two compartment model. The distribution ratio and ID% of nine viscera or tissue were measured and compared with other until the 40th day.

RESULTAfter oral administration, AA-I was rapidly absorbed into the blood and reached its peak at 30 minutes and lasted till 90 minutes. AA-I concentration in the blood gradually declined afterwards. 24 hours later, only few AA-I could be detected. By the 10th day, 68.5% of AA-I presented as the binding type with serum albumin. Pharmacodynamic parameters were calculated as follows: Tmax 0.74 h, Cmax 0.92 microg x mL(-1), t1/2alpha 0.68 h, t1/2beta 20.46 h, V/F 87.39 mL, CL(s) 5.85 mL x h(-1) (0.10 mL x min(-1)). On the other hand, after oral administration AA-I was rapidly distributed to all the viscera or tissue, whose peak appeared in 5 minutes and the vallecula was from 24 to 48 hours. The distribution ratio of AA-I rose in the kidney after 24 hours, and it showed the highest level in the kidney and in the liver by the 4th day compared with other organs or tissue (P < 0.05). However, the distribution ratio of AA-I in the kidney became the most dominant one after the 30th and the 40th day compared with the others (P < 0.05).

CONCLUSIONAA-I is rapidly absorbed after oral administration in rats. Its distribution has the organ specificity, which is characterized as the possible partial metabolism in the liver and the accumulation in the kidney because of rather slower elimination. The characteristics may be related to the long term nephrotoxicity of AA-I.

Administration, Oral ; Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; administration & dosage ; pharmacokinetics ; pharmacology ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Metabolic Clearance Rate ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Tissue Distribution

OBJECTIVETo investigate the therapeutic effects of acupuncture combined with nerve block for treatment of lumbar disc herniation.

METHODSNinety cases of lumbar disc herniation were randomly divided into three groups: acupuncture combined with nerve block group, acupuncture group and nerve block group, 30 cases in each group. The acupuncture combined with nerve block group was treated with acupuncture combined with nerve block therapy, L2-L5 Jiaji (EX-B 2), Zhibian (BL 54), Huantiao (GB 30), Yanglingquan (GB 34) etc. were selected for acupuncture, affected nerve root, sciatic nerve or common peroneal nerve were selected for nerve block with anti-inflammation-analgesic injection; the acupuncture group was treated with acupuncture only; the nerve block group was treated with nerve block only. After 4 weeks of treatment, the visual analogue scale (VAS), Oswestry lumbar dysfunction index (ODI) and modified MacNab standard were compared to evaluate the therapeutic effects of three groups.

RESULTSThe VAS and ODI in all groups were significantly decreased after one week, two weeks and four weeks of treatment (all P<0.01); after one week of treatment, the scores of VAS and ODI in nerve block group and acupuncture combined with nerve block group were significantly lower than those of acupuncture group (P<0.05); after two weeks and four weeks of treatment, the scores of VAS and ODI in acupuncture combined with nerve block group were significantly lower than those of acupuncture group and nerve block group (P<0.05). The effective rate and excellent and good rate of the acupuncture combined with nerve block group were significantly higher than those of acupuncture group and nerve block group (both P<0.01).

CONCLUSIONThe nerve block therapy and acupuncture are effective methods for treatment of lumbar disc herniation, while it has a better effect when these two treatments are combined used.

Acupuncture Therapy ; Adult ; Analgesics ; administration & dosage ; Anti-Inflammatory Agents ; administration & dosage ; Female ; Humans ; Injections ; Intervertebral Disc Displacement ; drug therapy ; therapy ; Male ; Middle Aged ; Nerve Block ; Treatment Outcome

Aim To investigate the effects of astragalus polysaccharide (APS) on renal TGF-β1/Smads signa-ling pathway in rats with diabetes mellitus(DM). Methods Wistar rats were randomly divided into nor-mal group,DM group,APS low dose (APS-low) group and APS high dose (APS-high) group. Rats in APS-low group and APS-high group respectively received 200 and 400 mg·kg-1·d-1APS for 8 weeks. Con-centrations of fasting blood-glucose(FBG),blood urea nitrogen and creatinine,as well as urinary kidney injury molecule-1 (KIM-1) and osteopontin (OPN) were measured. Levels of TGF-β1,Smad2,phosphorylated Smad2 (p-Smad2),Smad3,phosphorylated Smad3 (p-Smad3),Smad7,matrix metalloproteinase (MMP) 2,MMP-9,tissue inhibitor of matrix metalloproteinases (TIMP)-1 and TIMP-2 were investigated. Results Compared to control group,DM group had higher levels of FBG,blood urea nitrogen,creatinine KIM-1,OPN, TGF-β1,Smad2,p-Smad2,Smad3,p-Smad3,TIMP-1 and TIMP-2,but lower levels of Smad7,MMP-2 and MMP-9. APS significantly decreased the levels of FBG,blood urea nitrogen,creatinine KIM-1 and OPN, as well as inhibited the activity of TGF-β1/Smads sig-naling pathway. Conclusion The renoprotective effects of APS might be associated with the inhibition of TGF-β1/Smads signaling pathway.