Aim Toinvestigatetheeffectsoftriptonot-erpene methyl ether ( TME ) , a diterpene derived from the medicinal plant Triptergium wilfordii, on human gastric cancer AGS cell proliferation inhibition and ap-optosisinducedinvitro.Methods MTTassaywas used for screening tumor spectrum and detecting the vi-ability of AGS cells and normal human gastric epitheli-al cells GES-1 . Cell morphology was observed by light microscopy and AO / EB staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. JC-1 staining and fluorescence probe DCFH-DA were em-ployed to detect the changes of mitochondrial mem-brane potential and reactive oxygen species ( ROS ) . The effect of inhibiting AGS clonogenic survival was as-sayed by the method of plate clone formation. Western blot was used to analyse the expression of caspase-3 , caspase-8,Bcl-2andBax.Results MTTresults showed that TME exhibited significantly higher cytotox-icity to gastric cancer AGS cell line than to noncancer-ous cell line GES-1. IC50 for AGS of 48 h treatment was 23 . 85 μmol · L-1 . TME significantly inhibited colony formation and caused morphological changes in AGS cells. Annexin V-FITC / PI double staining showed the apoptotic rate increased. DCFH-DA stai-ning showed TME resulted in an increase in intracellu-lar ROS levels. Mitochondrial membrane potential de-creased after TME treatment. Western blot results showed that TME increased the proportion of Bax /Bcl-2 , with the activation of caspase-8 and caspase-3 . The broad-spectrum caspase inhibitor z-VAD-fmk pre-treatment reduced the expression of caspase-8 and caspase-3. TME enabled AGS cell cycle arrest in G0/G1phase.Conclusion TMEpossessespotenttumor selected toxicity and can induce apoptosis of AGS cells through cell cycle arrest, which is associated with Bcl-2 protein family.

Objective To investigate the effects of galangin on gene expressions of Nrf2 andγ-GCS in oxidative damage of A375 cell;To discuss its protective mechanism for anti-oxidative damage. Methods A375 melanoma cells were induced oxidative stress to establish oxidative damage model by 700μmol/L H2O2. The study was divided into normal group, model group, positive medicine group and high-, medium-, and low-dose galangin groups. All administration groups were given relevant medicine for cultivation. Cell viability was detected by MTT;ROS content was detected by ELISA;the gene expressions of Nrf2 andγ-GCS were detected by RT-PCR.Results Compared with normal group, cell viability decreased significantly;ROS content increased significantly;the gene expressions of Nrf2 andγ-GCS decreased significantly in the model group (P<0.05,P<0.01). Compared with model group, cell viability increased, ROS content decreased, the gene expressions of Nrf2 andγ-GCS increased significantly in all administration groups (P<0.05,P<0.01). Conclusion Galangin may activate Nrf2 signal path to realize the protective effect on A375 cellular oxidation damage through upregulating the expressions of Nrf2 andγ-GCS to promote the integration of Nrf2 and antioxidant response element and relevant regulatory enzymes.

ObjectiveTo study the effect of improving communication methods on decreasing the depression of pstients with stroke dysphasia.Methods16 patients with stroke dysphasia communicated with carton cards,body signal.All the patients were assessed by Hamilton Depression Scale(HAMD) in admission day, one week and two weeks later.ResultsAfter two weeks, the patients' depressive level was decreased from(24.2±8.4) to(13.4±6.7)(P<0.001), the symptom of depression reduced 81.2%.ConclusionsImproving communication methods can decrease the depressive level of the patients with stroke dysphasia.

Objective: To investigate the treatment efficacy of radiofrequency ablation (RFA) of hepatic tumors and the relevant complications. Methods: A total of 343 patients with 778 hepatic tumors underwent ultrasound-guided RFA (582 procedures). There were 212 cases of hepatic cellular carcinoma (HCC) with 448 tumors, and the average largest diameter was 4.0 cm.Of all the patients, 63 (29.7%) were in the stage of Ⅰ-Ⅱ(UICC Systems) and 149 (70.3%) in stage of Ⅲ-Ⅳ(including 43 patients with tumor recurrence after surgical resection). There were 131 cases of metastatic liver carcinoma (MLC), with 330 metastases in the liver, the average diameter was 3.9 cm, and the liver metastases of 91 patients (69.5%) came from gastrointestinal tract. The patients were treated using the relatively standard protocol. Crucial attention must be paid to monitoring the abnormal changes in ultrasound images as well as the vital signs of the patients to find the possible hemorrhage and peripheral structure injury in time. The tumors were considered ablated successfully if no viability was found on enhanced CT within 24 hours or 1 month after RFA. The patients were followed up for 2 -62 months.Results: The ablation success rate for HCC was 95.5% (428/448 tumors), and the rate for MLC was 96.4% (318/330 tumors). The local tumor recurrence rates for HCC and MLC were 8.5 % (38/448 tumors) and 11.8 % (39/330 tumors), respectively. A total of 138 patients (40.2%) underwent repeated ablations for 2-11 times because of tumor recurrence or metastasis. The first,second and third years survival rates were 87.7% , 67.4% and 56.8% for HCC patients, 81.6%, 50.8% and 27.2% for MLC patients,respectively. The survival rate from 63 early-stage HCC patients were 92.9%,82.8% and 74.5%, respectively. The major complication rate in this study was 2.4 % (14 of 582 procedures).The complications which consisted of mechanical and thermal injuries usually occurred during or shortly after the RFA treatment. There were 5 hemorrhages, 1 colon perforation, 5 injuries of adjacent structures, 2 bile leakages and 1 skin burn. Conclusion: RFA, as a minimally invasive local treatment, has become an effective and relatively safety alternative for the patients of hepatic tumors, even of advanced live tumor, tumor recurrence, liver metastases which are unresectable or difficult to treat with traditional therapies. Knowledge about possible complications and their control may increase the treatment efficacy and help to promote the use of RFA technique.

The gpr81 was amplified by polymerase chain reaction (PCR) using human fetus kidney cDNA and whole blood genome DNA as template, respectively. The expression profile of gpr81 in human fetus was analyzed by RT-PCR and the result indicated GPR81 mRNA was most abundant in fetus liver and heart. In addition, the deduced amino acid of GPR81 was compared with other related molecules by Clustal w/x software, and a molecular phylogenetic tree was constructed with Treeview software. It was showed that GPR81 had the highest homology with nicotinic acid receptor in amino acids. After sequence identification, gpr81 was inserted into the plasmid pcDNA3. 1 (-)/his-mycA and then transfected into Chinese hamster ovary cell (CHO-K1). With the selection of G418, an engineered cell line which could stably express gpr81 was obtained by the indication of RT-PCR and Western-blot detection. The establishment of the cell line will serve as means for further study of GPR81.
Amino Acid Sequence ; Animals ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Fetus ; metabolism ; Gene Expression Profiling ; Humans ; Molecular Sequence Data ; Phylogeny ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription, Genetic ; Transfection

Little is known about the distribution of Rhodococcus equi in the soil environment of native horses in China. One hundred and eight soil samples were collected from native-horse farms in the Hulun Beier grasslands of eastern Mongolia, the Xilin Goler grasslands of southern Mongolia, and Tongliao city in Inner Mongolia of China for investigating the distribution of R. equi in these regions. The isolation rates of R. equi from soil samples from the Hulun Beier and Xilin Goler grasslands ranged from 25.9% to 30.0%. In contrast, isolation rates from soil samples from Tongliao city was as high as 82.3% and the mean number of R. equi in soil samples from Tongliao city was 10 times more than those of samples from the grasslands. The 488 isolates were examined using PCR for the presence of genes that encode virulence-associated 15 000-17 000 antigen protein (VapA) and the 20 000 antigen protein (VapB). All isolates were negative for virulence-associated proteins. Plasmid profiles of these avirulent isolates showed that cryptic plasmids of various sizes were present with an incidence of 13.3% to 21.5%. The results of the present study contrast with those of our recent study, in which we reported that R. equi was absent from Mongolian horses in Ulaanbaatar, Mongolia. It is suggested that the difference between the results of these two studies is due to the mobile pasturing system in Mongolia and nonmobile pasturing system in Inner Mongolia.

AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.

METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.

RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.

CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance

BACKGROUNDInflammation is a major cause of restenosis after coronary stenting. Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule that plays a key role in the tight adhesion between leukocytes and vascular endothelium. The object of this study was to investigate the association between the K469E polymorphism of the ICAM-1 gene and restenosis after coronary stenting in North Chinese population.

METHODSThe ICAM-1 K469E polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism method in 124 patients who had undergone coronary stenting and coronary angiography at least 3 months earlier. Information on clinical risk factors and procedure-related data were also collected.

RESULTSOf 124 enrolled patients in total, there were 72 cases of in-stent restenosis. The restenosis rate in this population was 58.1%. The frequencies of the three possible genotypes of the ICAM-1 K469E polymorphism were: KK genotype 50.8%, EE genotype 41.9%, and EK genotype 41.9%. Among restenosis patients, the frequency of the KK genotype was 58.3% and the frequency of E allele carriers was 41.7%. Among non-restenosis patients, the frequency of the KK genotype was 40.4%, and the frequency of E allele carriers was 59.6%. The distribution of these two genotype groups between restenosis and non-restenosis patients was significantly different (P = 0.049). Using multivariate logistic regression, the difference between the two groups was more apparent. The odds ratio of KK homozygotes vs E allele carriers was 2.6, with 95% confidence interval 1.2 - 5.8 (P = 0.018). After grading of risk factors, we found that the KK genotype was a stronger predictor of in-stent restenosis in obesity or hyperlipemia patients, with an odds ratio of 9.3 and 3.7, respectively (P < 0.05).

CONCLUSIONIn our study population, KK homozygotes of the ICAM-1 codon 469 mutation had a higher risk of restenosis after coronary stenting, especially in the case of obese or hyperlipemia patients.

Asian Continental Ancestry Group ; genetics ; China ; Codon ; Coronary Restenosis ; genetics ; Female ; Genotype ; Humans ; Hyperlipidemias ; complications ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Middle Aged ; Obesity ; complications ; Polymorphism, Genetic ; Stents

OBJECTIVETo establish a HPLC method for determining acetoside in rat plasma and to investigate the pharmacokinetic characteristics of acetoside in rats.

METHODSix rats were orally administered with 150 mg x kg(-1) acetoside and their blood samples were collected at different time points. The plasma concentration of acetoside was determined by reserved HPLC, and the pharmacokinetic parameters were calculated by DAS 2.0 software.

RESULTThe regression equation of acetoside in rats plasma was Y = 3.509 8X-0.096 8 (r = 0.996 8), which showed a good linear relation at 0.125-2.5 mg x L(-1). The method showed a recovery of more than 85%, and both inter-day and intra-day RSDs were less than 15%. After the oral administration of 150 mg x kg(-1) acetoside, the concentration-time curves of acetoside were expressed in a open two-compartment model. The main pharmacokinetics parameters of T(max), C(max), t(1/2alpha), t(1/2beta), AUC(0-t), AUC(0-infinity), CL/F, V/F and K(a) were respectively 0.36 h, 1.126 mg x L(-1), 0.759, 4.842 h, 3.134, 3.766 mg x h x L(-), 87.089 L x h(t) x kg(-1), 207.704 L x kg(-1) and 6.345 h(-1) respectively.

CONCLUSIONIt is first time to establish such a HPLC method to determine the concentration of acetoside in plasma. The method is so highly specified and sensitive that it can ble used in quantitative analysis in vivo on acetoside.

Animals ; Chromatography, High Pressure Liquid ; Female ; Glucosides ; chemistry ; pharmacokinetics ; Male ; Phenols ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cricetinae ; Cricetulus ; Gene Expression Profiling ; Green Fluorescent Proteins ; genetics ; Humans ; Molecular Sequence Data ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Tissue Distribution ; Transfection