The chemical constituents of Vaccinium bracteatum were studied by means of macroporous resin, ODS column chromatography and preparative HPLC. Eleven compounds were isolated from this plant. By using ESI-MS and NMR, the structures of the eleven compounds were determined as 10-O-trans-p-coumaroyl-6alpha-hydroxyl-dihydromonotropein (1), 10-O-cis-p-coumaroyl -6alpha-hydroxyl-dihydromonotropein (2), vaccinoside (3), 10-O-cis-p-coumaroyl monotropein (4), isolariciresinol-9-O-beta-D-xyloside (5), tectoridin (6), vicenin-3 (7), quercetin-3-O-alpha-L-rhamnoside (8), quercetin-3-O-alpha-L-arabinopyranoside (9), quercetin-3-O-beta-D-galactopyranoside (10), and quercetin-3-O-beta-D-glucuronide (11), respectively. Compounds 1 and 2 are new, and compounds 4, 6 and 7 are isolated from the genus Vaccinium for the first time.
Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Vaccinium ; chemistry

The PDB culture medium was selected to ferment the endophyte strain, and the secondary metabolites of endophytic fungi Penicillium polonicum were studied. Combined application of Sephadex LH-20, ODS and HPLC chromatographies over the ethyl acetate extract of the fermented culture led to the isolation of 6 compounds. By spectral methods, the structures were elucidated as [3, 5-dihydroxy-2-(7-hydroxy-octanoyl)]-ethylphenylacetate (1), (3, 5-dihydroxy-2- octanoyl)-ethyl phenylacetate (2), (5, 7-di- hydroxy-9-heptyl)-isobenzo pyran-3-one (3), 3-(hydroxymethyl) 4-(1E)-1- propen-1-yl-(1R, 2S, 5R, 6S)-7-oxabicyclo [4.1.0] hept-3-ene-2, 5-diol (4), (E)-2-methoxy-3-(prop-1-enyl) phenol (5) and p-hydroxylphenylethanol (6).
Biological Factors ; chemistry ; metabolism ; Endophytes ; chemistry ; isolation & purification ; metabolism ; Fabaceae ; microbiology ; Fermentation ; Penicillium ; chemistry ; isolation & purification ; metabolism ; Secondary Metabolism

Objective To detect the expression of humar telomerase reverse transcriptase (hTERT) and telomerase activity in HeLa,MCF-7,SMMC7721 PC-3m and U2OS cell lines. Methods The techniques of immunochemistry and TRAP-ELISA were employed to detect the expression of hTERT and telomerase activity in different tumor cell lines.Results The positive rate of hTERT in HeLa cells(93.75%?3.10%)was significantly higher than that in U2OS cells(2.75%?0.96%),besides the other three cell lines showed an positive rates of hTERT in MCF-7(92.50%?2.65%),SMMC7721 (53.75%?2.22%)and PC-3m(23.50%?2.89%); Meanwhile,the telomerase activity of HeLa cells (94.58%?3.49%) was also much higher than that of U2OS(3.02%?0.43%),likewise the telomerase activities of the other three cell lines (MCF-7,SMMC7721,PC-3m) were 73.90%?4.50%,66.67%?3.35% and 50.62%?1.96%, respectively. Conclusion The expression of hTERT and telomerase activity show obvious differences among five tumor cell lines,suggesting that telomerase inhibitors cannot effect on all the tumor cell lines.

Objective: To investigate the effect of aqueous extract of Radix et Rhizome Rhodiolae on expressions of HIF-1?, HIF-1? and VEGF in human umbilical vein endothelial cells(HUVECs) exposed to hypoxia. Methods: The effect of different treatment time and concentrations of aqueous extract of Radix et Rhizome Rhodiolae on cellular proliferation were determined with XTT colorimetric assay. The mRNA and protein expressions of HIF-1?, HIF-1? and VEGF were analyzed respectively by quantitative real-time polymerase chain reaction with SYBR Green I and Western blot. Three groups were studied: normoxia control group, hypoxia control group, Radix et Rhizome Rhodiolae treatment group. Results: The optimal condition of Radix et Rhizome Rhodiolae treatment were 24h and 10?g/mL. The protein levels of HIF-1?, HIF-1? and VEGF were elevated by hypoxia (P

Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.

Objective To investigate the role of PI_3 K/Akt signal pathway in Ephrin-Al gene mediated invasion,metastasis of Huh-7 cells.Methods Western blot was used to test the protein expression of phosphatidylinositol 3-kinase(PI_3 K)and mitogen-activated protein kinase(MAPK)after Huh-7 cells were treated with Ephrin-A1/Fc fusion protein.According to the protein expression,LY294002 was used to block PI_3 K/Akt pathway specifically,then p-Akt protein expression,mobility and invasive ability of Huh-7 cells were examined.Results In Huh-7 cells actived by Ephrin-Al/Fc fusion protein,p-Akt expression was higher than that in control group(t=4.564,P＜0.05),but there was no difference of p-p38MAPK expression between Ephrin-Al/Fc fusion protein group and IgG/Fc fusion protein group(P＞0.05).PI_3 K/Akt pathway was specifically blocked by LY294002,the p-Akt protein expression decreased in Huh-7 cells,and the mobility and invasive ability mediated by Ephrin-Al in Huh-7 cells decreased(P＜0.05).Conclusions PI_3 K/Akt pathway effects an important role in mobility and invasive ability of Huh-7 cells mediated by Ephrin-A1.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Fluorescence ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; isolation & purification ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Tuberculosis, Pulmonary ; diagnosis ; microbiology ; Young Adult

Objective Toinvestigatethepossibilityandquantitativerangeofpepsinogen(PG)usedas the screening marker of gastric cancer by detecting serum pepsinogen level in different gastric mucous pathologic status.Method ThelevelofserumpepsinogenⅠ(PGⅠ)andpepsinogenⅡ(PGⅡ)bytimeresolvedfluoro-immunoassay(TRFIA)in 64 chronic atrophic gastritis patients,63 gastric mucous atypical hyperplasia patients, 67 gastric cancer patients and 20 healthy volunteers were defeeted ,and the ratio of PGR(PGⅠ/PGⅡ)was calculated.Then the three kinds of diseases were graded.The data was analyzed between groups and sub-groups.Result ①Compared with normal control group,the median PGⅠvalues were 1 24.01 ,91 .23 and 71 .23 respectively,which were all lower than that of healthy group (1 52.00).There were significant differ-ences(Z=-2.52,P=0.01 7 0;Z=-3.42,P=0.001 4;Z=-3.57,P=0.000 9).The median PGR values were 7.61 ,5.21 and 4.32 respectively,which were also lower than that of healthy group,the differences were significant(Z=-2.98,P=0.002 9;Z=-3.1 7,P=0.000 2;Z=-2.89,P=0.000 1 ).The PGⅡlevel of these diseases were not significantly different with control group.②The serum PGⅠlevel of gastric mucous atypical hyperplasia and gastric cancer were reduced significantly in contrast with atrophic gastritis (Z =-3.42,P =0.001 4;Z=-3.62,P=0.000 9);the levels of PGⅡand PGR were varied without significance (P>0.05 );③The levels of PGⅠamong atrophy gastritis and gastric cancer subgroup have no significant difference(χ2 =2.86,P=0.41 4 3;χ2 =1 .67,P=0.1 36 8).But the level of PGⅠwas significantly different in gastric atypical hyperplasia(χ2 =0.83,P=0.043 0).It decreased in light and medium grade dysplasia and went up in severe grade dysplasia.The levels of PGⅡ and PGR were varied without significance(P>0.05).④The areas under the ROC curves performed by the PGⅠ and PGR from normal control group and atypical hyperplasia group were 0.782 and 0.831 respectively;The sensitivity and specificity of PGⅠ≤72.1 2 μg/L and PGR≤4.32 for gastric dysplasia were 89.48%and 76.31%respectively.Conclusion ①The level of PGⅠand PGR were decreased along with the seriousness of gastric pathological changes and probably regarded as the screening markers of gastric mucous malignant transformation.②Serum pepsinogen level is closely correlated with gastric precancerous lesion,PGI≤72.1 2 μg/L and PGR≤4.32 has better specificity and sensitivity for gastric atypical hyperplasia in this area.

Objective To assess the variation in patient setup corrections for three different anatomic treat-ment sites using daily pretreatment megavoltage CT(MVCT)in helical tomotherapy,and to analyze alternative refer-ence margins for specific tumor site.Methods Sixty patients treated for three anatomical sites on helical tomotherapy were analyzed.Daily MVCT was carried out for registration and setup corrections before each treatment fraction.Setup errors and rotational setup corrections from 587 head and neck,500 thoracic,371 abdomen and gynecology MCVT imaging were input to statistical analysis.Incidences of three dimensional vector error lengths were calculated for each anatomic site.Overall distribution histograms of the three -dimensionally error was presented using GraphPad Prism 5.The distributions of systematic and random setup errors were quantitative analyzed and the additional margins required were also taken into account.Results 1 458 MVCT scans were implemented for sixty patients.Head and neck had lower frequencies of translational setup errors than others.Frequency of at least 3mm three -dimensional setup errors for head and neck,thoracic,abdomen and gynecology was 55.3%,70.8%,79.8%,respectively.This fre-quency decreased to 17.5%,40.6%,47.2% if 3D vector distance ≥5mm was scored.Overall systematic errors ranged from -1.4mm to 2.7mm,abdomen and gynecology had the largest setup errors in the vertical direction which was statistically significant(χ2 =19.3,P <0.05).The suggested margins should be increased by 4 -7 mm in three -dimensional direction for head and neck,9 -14mm for thoracic,10 -17mm for abdomen and gynecology.Conclusion Differences in setup corrections are perceived between head and neck,thoracic,abdomen and gynecology.The accura-cy of patient positioning can be improved if pre -treatment daily MVCT scans are put into use.Results from setup cor-rection can provide evidence for tumor treatment margin and improve the accuracy of regular radiotherapy.

Objective To evaluate the efficacy and safety of short-term restriction of dietary protein intake (DPI) supplemented with α-keto acids on chronic hepatitis B patients complicated with chronic kidney diseases (CKD). Methods A prospective randomized controlled trial was carried out.Seventeen chronic hepatitis B patients with CKD were randomized to either low DPI with α-keto acid-supplemented (sLP) or low DPI (LP) group for 3 months.Low-protein diet (LPD) was individualized with total energy intake 125.52-146.44 kJ·kg-1 ·d-1,and protein intake of 0.6-0.8 g·kg-1·d-1.α-keto acid was supplied in a dosage of 0.1 g·kg-1·d-1.Nutritional indexes were recorded and other clinical indexes were measured to evaluate the efficacy and safety respectively. Results The urine protein excretion level and microalbuminuria were significantly decreased at the end of the observation period in the sLP group compared to the basal value and the LP group [24 h urine protein:baseline (4.52±1.74) g,the 1st month (3.19±1.52) g,the 2nd month (2.19±1.1) g,the 3rd month (1.64±0.77) g,P＜0.05; microalbuminyria:baseline (2855.43±248.03) mg/L,the 1st month (2157.14±218.15) mg/L,the 2nd month (1681.57±146.18) mg/L,the 3rd month (924.29±83.33) mg/L,P＜0.05].No significant difference was found in Scr and eGFR.Nutritional indexes (SGA,serume albumin) were significantly higher at the end of 3 months in the sLP group (P＜0.05).No obvious side-effect occurred. Conclusions Short-term restriction of DPI is safe,and when combined with α-keto acids,can increase serum protein and decrease urine protein excretion in chronic hepatitis B patients complicated with CKD without significant sideeffect.