The change of kirenol, darutigenol and darutoside in Siegesbeckia and its first to ninth processed products were studied, and the ten fingerprints were compared, which provided the experimental basis for the study of Siegesbeckia processing tech- nology. The samples were analysed by HPLC on a SunFire-C18 column (4.6 mm x 150 mm, 5 μm) with gradient elution of acetonitrile (0.1% formic acid)-water (0.1% formic acid) at a flow rate of 1.0 mL x min(-1). Column temperaturewas 30 °C and the detected wavelength was 215, 320 nm. The calibration curves of kirenol, darutigenol and darutoside were linear in the range of 2.180-26.16, 2.900-34.80, and 1.012-6.072 mg x L(-1), respectively, and the average recoveries were 96.4%, 97.2% and 96.3% wit RSD 2.2%, 1.7% and 2.4%. This method was simple, the result was stable and had good repeatability, recovery and precision. The re- sult was the basis of the chemical contents variation in the processing of Siegesbeckia Herbs and further clarifying the effect of the changing.
Asteraceae ; chemistry ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Temperature

OBJECTIVETo observe targeted expression of recombinant lentivirus-mediated (Lv)-hTERTp-TK and Lv-hTERTp-tumstatin in HepG2 cells, and explore the inhibitory effect of their combination on HepG2 cells both in vitro and in vivo.

METHODSLv-hTERTp-TK and Lv-hTERTptumstatin were used to infect HepG2 and L02 cells at different MOIs. Transfection efficiency was observed by fluorescence microscopy. Expression of TK and tumstatin mRNA was detected by reverse-transcriptase PCR. Proliferation and apoptosis were detected by MTT and flow cytometry, respectively. The HepG2 cells were examined by real time-PCR and western blotting to determine expression level of bcl-2 and VEGF mRNA and protein.A murine hepatocellular carcinoma model was established by injecting 1 * 10(7) HepG2 cells into 30 BALB/c nude mice. The modeled mice were randomly divided into a control group, mock group, Lv-hTERTp-tumstatin group, Lv-hTERTp-TK group, and combination group for four weeks of injections at regular intervals of PBS, Lv-hTERTp-null, Lv-hTERTp-tumstatin, Lv-hTERTp-TK, and Lv-hTERTp-tumstatin plus Lv-hTERTp-TK, respectively.Changes in tumor volume and weight, and cell morphology of tumor and major organs, were assessed by hematoxylin-eosin staining.Microvascular density of tumor tissue and cell apoptosis were assessed by immunohistochemical and TUNEL staining, respectively.

RESULTSThe Lv-infected HepG2 cells, and not the Lv-infected L02 cells, expressed TK and tumstatin. Lv-hTERTp-TK and Lv-hTERTp-tumstatin, alone or in combination, inhibited proliferation and increased apoptosis of the HepG2 cells, but the combination was more effective than either alone (P less than 0.05). None of the treatments affected proliferation or apoptosis of the L02 cells (P more than 0.05). The combination also led to a greater reduction of bcl-2 and VEGF than either alone (all, P less than 0.05). Tumor growth was significantly inhibited by the combination (P less than 0.05). In vivo, the combination treatment induced the greatest amount of apoptosis of the HepG2 cells. Cell morphology of major organs such as liver, spleen and kidney were similar to the control group. The combination also produced the most significant effect on tumor microvascular density (P less than 0.05) and the highest apoptosis index (P less than 0.05).

CONCLUSIONThe HTERT promoter can induce targeted expression of TK and tumstatin in HepG2 cells. Lv-hTERTp-TK combined with Lv-hTERTp-tumstatin can significantly inhibit proliferation and induce apoptosis of human HepG2 cells in vitro and in vivo, which may be related to down-regulation ofbcl-2 and VEGF.

Animals ; Apoptosis ; Autoantigens ; genetics ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Collagen Type IV ; genetics ; Down-Regulation ; Flow Cytometry ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lentivirus ; Liver Neoplasms ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Telomerase ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

This research use "3414" fertilizer effect experiments to handle zinc, boron and molybdenum trace element fertilizer, determined the dry matter accumulation and content of saikosaponion a and d, to investigate the different ratio of zinc, boron and molybdenum on yield and saikosaponin a, saikosaponin d contents of Bupleurum chinense. Found The suitable ratio of zinc, boron and molybdenum play an active role on dry matter accumulation and distribution, the treatment Zn2B2Mo3 is the best one to promote the dry matter accumulation and transfer to the underground part; in a certain range, only use zinc or molybdenum can promote the yield of B. chinense, the yield of treatment Zn2B2Mo1 is the highest one. According to the results of regression analysis: in accordance with Zn 48.45 g x hm(-2), B 355.05 g x hm(-2), Mo 86.40 g x hm(-2), can obtain the yield with 3313.05 kg x hm(-2); the treatment Zn2BMo2 is most effective to promote the total saikosaponin a and d accumulated, according to the results of regression analysis: in accordance with Zn 36.15 g x hm(-2), B 343.05 g x hm(-2), Mo 106.35 g x hm(-2), the content of total saikosaponin a and d can reach 1.23%. This research first discovered the suitable ratio of zinc, boron and molybdenum can promote the yield and saikosaponin a, saikosaponin d contents on B. chinense.
Boron ; metabolism ; Bupleurum ; metabolism ; Fertilizers ; Molybdenum ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Roots ; metabolism ; Saponins ; metabolism ; Trace Elements ; metabolism ; Zinc ; metabolism

Bacteria ; chemistry ; Microbial Sensitivity Tests ; Microbiological Techniques ; Reagent Kits, Diagnostic

In order to select high quality and suitable Bupleuri Radix varieties in Qingchuan, and establish a new comprehensive method to evaluation the quality of Bupleuri Radix, 12 characters of 14 samples were evaluated by DTOPSIS and grey related degree. The results showed that varieties No. 8 and No. 10 had high quality. DTOPSIS and grey related degree gave the uniformity result, and the biggest difference of value of Ci in DTOPSIS method was 46. 33% , but the biggest difference of the weighting correlation number( r (i)) in grey related degree was only 13.10%. The DTOPSIS combined with grey related degree can evaluate the quality of Bupleuri Radix comprehensively and objectively.
Bupleurum ; chemistry ; China ; Drugs, Chinese Herbal ; chemistry ; supply & distribution ; Quality Control ; Statistics as Topic ; methods

Objective To explore the expression of hypoxia-inducible factor-1?(HIF-1?) protein and its mRNA in cultured cortical neurons after hypoxia,ischemia or hypoxia-ischemia(HI) and explore the possibilities of HIF-1? gene therapy in HI neurons.Methods The in vitro models of HI,pure hypoxia and pure ischemia were established using embryonic day 16-18 rats cortical neurons.Immunohistochemical and in-situ hybridization were performed to examine the expression of HIF-1? protein and its mRNA at different reperfusion time points in neurons.Results The expression of HIF-1? protein was very week in normoxic cultured neurons,but was up-regulated while treated with hypoxia and(or) ischemia.HIF-1? expression reached peak at 4 to 8 h after reperfusion with HI,which were statistically significant higher than that at other time points(Pa=0),and decreased gradually at 12 h.Furthermore,HIF-1? protein expression was significantly higher in HI group compared with that in the pure hypoxia or ischemia group(Pa=0).HIF-1? mRNA reached peak immediately after HI,decreased gradually at 2 h,and returned to the baseline at 8 h after reperfusion.Conclusions HIF-1? expression on cortical neurons is regulated differently with hypoxia,ischemia or HI treatment,HIF-1? gene therapy for HI neurons maybe a useful method in the future studies.

OBJECTIVETo evaluate the feasibility of angiography combined with transthoracic echocardiography (TEE) as a modified management of the transcatheter occlusion of patent ductus arteriosus (PDA).

METHODSForty children with PDA were randomly divided into two groups (n=20 each): observed and control. The control group accepted traditional transcatheter occlusion, and the observed group received a modified management (angiography combined with TEE). The children in the observed group were monitored by realtime TTE.

RESULTSA complete occlusion was acquired by one occlusion operation in each child in the observed group. The TTE demonstrated that the occlusion device was in place, and that the blood flow velocities in the left and right pulmonary artery and the descending aorta were in normal ranges. There were shorter X-ray exposure time, shorter recovering time and less ICU stay time in the observed group than in the control group. The complications associated with blood vessel puncturation occurred in four children from the control group, but none of the observed group had the complications. The total hospitalization cost in the observed group was less than in the control group.

CONCLUSIONSAngiography combined with TEE as a modified management of the transcatheter occlusion of PDA is recommended.

Adolescent ; Cardiac Catheterization ; Cardiac Surgical Procedures ; methods ; Child ; Child, Preschool ; Ductus Arteriosus ; diagnostic imaging ; Ductus Arteriosus, Patent ; diagnostic imaging ; surgery ; Echocardiography ; Humans ; Infant ; Radiography

In this paper, duloxetine was chosen as the lead compound. The pharmacophores with 5-HT(1A) antagonism activity were used to replace the naphthyl of duloxetine. A series of duloxetine derivatives had been designed and synthesized and whose structures were confirmed with elemental analysis, MS and H NMR. All synthesized compounds were tested by tail suspension test and forced swimming test in vivo. The test results revealed that most of the compounds have shown better activity than duloxetine at the same dosage. Some of them are worth to be studied further.
Animals ; Antidepressive Agents ; chemical synthesis ; chemistry ; pharmacology ; Duloxetine Hydrochloride ; Hindlimb Suspension ; Male ; Mice ; Mice, Inbred ICR ; Molecular Structure ; Serotonin 5-HT1 Receptor Antagonists ; pharmacology ; Structure-Activity Relationship ; Swimming ; Thiophenes ; chemical synthesis ; chemistry ; pharmacology

Acetylcholine ; metabolism ; Animals ; Arcuate Nucleus of Hypothalamus ; metabolism ; Dopamine ; metabolism ; Epinephrine ; metabolism ; Female ; Neurotransmitter Agents ; metabolism ; Norepinephrine ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Synaptic Transmission

HPLC-ELSD was applied to explore the absorption mechanism of pulchinenosides (B3, BD, B7, B10, B11) in rats. The experimental results showed that the absorption rate constant, Ka value (B3, BD) and Permeability coefficient, Peff value (B3, B7) displayed significant difference (P <0.05) in various intestinal segments, The Ka value and Peff value of PRS was different from each other with the highest absorption in duodenum (duodenum > jejunum > colon > ileum); The PRS displayed excessive satuation as the concentration increased over 0.05-2.5 g · L(-1). There were no obvious linear correlations between Peff values and concentrations in duodenum (0.6007 ≤ R2 ≤ 0.7727); Ka and Peff value declined when the PRS was perfused with P-glycoprotein promoter digoxin, on the other hand, inclined when perfused with P-glycoprotein inhibitor verapamil with significant difference among PRS B3, BD, B7, B11 (P <0.05). All the above results demonstrated that B3, BD, B7 were greatly influenced by absorption sites, duodenum was the main absorption site; PRS didn't entirely transported in a concentration dependent manner, and the transporter-protein involved the transportation, so the intestinal absorption of the five pulchinenosides was not entirely passive diffusion; and PRS might be the substrates of P-glycoprotein.
ATP-Binding Cassette, Sub-Family B, Member 1 ; physiology ; Animals ; Intestinal Absorption ; Male ; Oleanolic Acid ; pharmacokinetics ; Pulsatilla ; chemistry ; Rats ; Rats, Wistar ; Saponins ; pharmacokinetics