Objective To detect the interaction domains' gene mutations of KEAP1 and NRF2 in non-small-cell lung cancer cell lines,and to analyze the significance of these mutations on the study of lung cancer cell lines.Methods Six non-small-cell lung cancer cell lines were used as the research materials.The 2nd to 6th exons of KEAP1 and the coding sequences of DLG and ETGE motif were amplified by PCR,and the products were used for gene sequencing analysis.Obtained gene sequences were analyzed using NCBI databases to get the base locations of gene mutations,as well as the subsequent amino acid sequence changes.Meanwhile,the relative intracellular reactive oxygen species (ROS) concentrations in the tested cell lines were detected and used in the analysis of abnormal NRF2 transcriptional activity in lung cancer cell.Results Four mutations were detected in the 4th exon of KEAP1 gene from all the 6 cancer cell lines,several other missense mutations were also investigated in the 3rd exon of KEAP1 from some cancer cell lines.Multiple genetic variations were found in all the NRF2-ETGE motif-encoding sequences of the 6 cancer cell lines.All 6 cancer cell lines were found to have higher ROS concentration than the lung germ cell line.Conclusion Lung cancer cells generally contain high levels of ROS as well as gene mutations in KEAP1 and NRF2 genes which lead to abnormal transcriptional activity of NRF2 in lung cancer cell lines.

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

Objective To observe the curative effect of low melocular weight heparin(LMWH) on the hypercoagulability in acute Kawasaki disease(KD).Methods Forty-six patients were diagnosed KD.Twenty-two cases out of all KD patients whose serum concentration of whether platelet(PLT) or fibrinogen(FIB) was significantly increased or who were found thrombus in their coronary artery by ultrasonic Doppler were treated with LMWH by subcutaneous injection once every day for 7-10 days.All the patients were divided into 2 groups accor-ding to whether using LMWH or not:H group(using LMWH) and NH group(no using LMWH).It were detected before and after treatment that included thrombin time(TT),activated partial thromboplastin time(APTT),international normalized ratio(INR),FIB,plasma mucosity,erythrocyte sedimentation rate(ESR),hematocrit(HCT) and situation of haemorrhage.Results 1.Before treatment,PLT and FIB of patients in H group were significantly higher than those in NH group(Pa

Flocculation of fermented broth with 1,3 -propanediol using component B of natural clarifier-Ⅱ was studied. Firstly, single factor tests and orthogonal tests were carried out. The results showed the main factors which influenced the flocculation and the optimal conditions were: pH6.0, flocculant B 0.01g/L , NaCl 0g/L, and the FR was 95.97% . The following results by filtration experiments, compared with the sample without pretreatment, indicated that the filtration speed of the flocculated sample was improved dramatically, and the weights of net and dry filter cakes were increased by 41.13% and 51.88% , respectively. Electrodialysis experiments showed that flocculation could accelerate desalting process, and could take the place of centrifugal pretreatment when it combined with the filtration.

Objective To evaluate the diagnostic value of physical examination,digital mammography and color Dopplor ultrasonography for breast masses.Methods The materials of clinic,digital mammography and color Dopplor ultrasonogrpahy(US)in 92 women(raged from 18～80 years old in age,with average of 49.8 years old)with breast masses confirmed by pathology were retrospectively analysed,including benign in 43 and malignant in 49.Digital mammographic findings including the location,shape,margins,calcifications and the lymph node of axillary of the breast lesions,and color Dopplor ultrasonographic findings including mainly the characteristics of lesions on two-dimensional sonography,the distribution of the internal and peripheral vessels of the lesions were observed,the physical examination mainly observed the skin and nipple,and the margins,texture and movement of the lesions.Results In diagnosing breast masses,the sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 85.71%,79.07%,82.35%,82.93% and 82.61%,respectively with digital mammography,79.59%,83,72%,84.78%,78.26% and 81.52%,respectively with US,71.42%,76.74%,77.78%,70.21%,73.91%,respectively with physical examination,while were 91.84%,93.02%,93.75%,93.3% and 90.91%,respectively in combination with three methods,which were significantly higher than that of each single examination(P＜0.05),but there were no differences between each examination(P＞0.05).Conclusion It can improve the diagnosing accuracy of breast masses by suitable combination of three of digital mammography,color Dopplor ultrasonography and physical examination.

OBJECTIVETo study the influence of different drying methods on the quality of Schisandrae Chinensis Fructus and thus provide useful reference for its proper drying methods.

METHODSchisandrae Chinensis Fructus was processed by eight drying methods including vacuum freeze drying, natural drying in the shade, drying in the sun, oven drying and vacuum drying under different temperature. The contents of the functional ingredients includes chisandrin, gomisin D, gomisin J, schisandrol B, angeloylgomisin H, angeloylgomisin Q, gomisin G, schisantherin A, deoxyschisandrin, schisandrin B, schisandrin C, 5-HMF, total aids and total sugars. The main components change after drying were analyzed by HPLC, ultraviolet spectrophotometry and potentiometric titration. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Schisandrae Chinensis Fructus.

RESULTAll these results are in accordance with the requirements of Chinese Pharmacopoeia published in 2010, the contents of schisandrin and total eleven lignans were the highest using vacuum drying, and 5-HMF were the lower, oven drying made little difference but with lower schisandrin and higher 5-HMF as the heat increased.

CONCLUSIONDifferent drying methods have significant influence on the quality of Schisandrae Chinensis Fructus. Oven drying under 5°C should be adopted to substitute drying in the sun according to the China Pharmacopoeia published in 2010 for Schisandrae Chinensis Fructus by comprehensive analysis of the cost, content and practicality.

Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Schisandra ; chemistry ; Temperature

OBJECTIVETo provide inspiration and ideas for clinical treatment of coronary heart disease (CHD) by data mining technology based frequency analysis and cluster analysis of medical records, prescriptions and herbs in treating CHD by distinguished veteran doctors of traditional Chinese Dedicine (TCM).

METHODSTotally 386 medical cases were retrieved from Wanfang Data, Chinese Scientific Journals Database (VIP medical information resources system, China National Knowledge Infrastructure (CNKI), and Typical Collections of Medical Cases by Contemporary Distinguished Veteran Doctors of Traditional Chinese Medicine. They input into database trimmed after unified standard. Medication laws of CHD by distinguished veteran doctors of TCM were analyzed using frequency analysis and cluster analysis, and so on.

RESULTSDistinguished veteran doctors of TCM frequently used top ten herbs in treatment of C D as Salvia miltiorrhiz , Ligusticum wallichii, Trichosanthes kirilowi, Pinellia ternat, Angelica sinensis, Poria coco stragalu , Panax ginseng, Allium macrostemon, and Radix Ophiopogonis. Cluster analysis summarized that there were 16 herb pairs commonly used, 7drug assemblies consisting of 3 herbs and 5 drug assemblies consisting of multiple herbs.

CONCLUSIONSDistinguished veteran doctors of TCM mainly used herbs assemblies capable for invigorating Pi to resolve phlegm, and promoting qi and activating blood circulation in treating CHD. Meanwhile, they concurrently used herbs combination of nourishing Xin and tranquilization, and regulating yin and yang.

China ; Cluster Analysis ; Coronary Artery Disease ; therapy ; Data Mining ; Databases, Factual ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional

The ginsenoside-Ro is sensitive to be hydrolyzed in an alkaline medium. This paper investigated the hydrolysis kinetics of ginsenoside-Ro under different pH and temperature values. The results showed ginsenoside-Ro in alkaline solution followed pseudo-first-order reaction. Hydrolysis kinetics of ginsenoside-Ro has not been reported previously. The hydrolysis rate was independent of initial concentration. On the basis of UFLC-MS/MS, NMR, as well as chemical evidence,the structure of hydrolyzate was assigned as 3-O- [beta-D-glucuronopyranosyl- (1 --> 2) -beta-D-glucopyranosyl] -oleanolic acid.
Drugs, Chinese Herbal ; chemistry ; Ginsenosides ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Kinetics ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.

OBJECTIVETo explore the relationship between expression of Fas, bcl-2 and apoptosis of cardiomyocytes in myocardial ischemia in rats.

METHODSMyocardial ischemia was experimentally induced by ligating the left coronary artery. The rate were killed from 10 minutes to 7 days after the operation. Apoptotic myocardial cells were detected by TUNEL method. The expression of Fas and bcl-2 was studied by ABC immunohistochemistry.

RESULTSCardiomyocytic apoptosis appeared from 3 to 36 hours after ischemia. This phenomenon however could not be detected by TUNEL method 7 days after ischemia. The expression of Fas could be detected by ABC immunohistochemistry from 3 hours to 7 days after ischemia, and the expression of bcl-2 from 10 minutes to 7 days. Cardiomyocytic apoptosis and Fas / bcl-2 expression appeared in different regions of myocardium: apoptosis in the ischemic regions, while Fas and bcl-2 expression in regions without obvious ischemia.

CONCLUSIONIn rats, Fas and bcl-2 may not directly regulate apoptosis of cardiomyocytes in case of myocardial ischemia.

Animals ; Apoptosis ; Male ; Myocardial Infarction ; metabolism ; pathology ; Myocytes, Cardiac ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism