Objective To investigate the roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells.Methods Different phenotypes of dCK plasmids were transfected into MCF-7 cells by liposome transfection,including dCK-Vector,dCK-WT (wild type),dCK-S74A (non-phosphorylation) and dCK-S74E (hyper-phosphorylation).All these cells were irradiated by 0,2,4,6,8 Gy X-rays,respectively.The transcriptional and translational level of dCK were detected with real time-PCR and Western blot,respectively.Radiosensitivity was analyzed using cell counting kit (CCK-8) and colony formation assays.Monodansylcadaverine staining (MDC) and flow cytometry were used to detect autophagy and apoptosis,respectively.Results Four phenotypes of dCK cell models were established successfully.After irradiation,the cell viabilities of MCF-7 and dCK-Vector decreased significantly as compared with mock group (t =14.469 and 9.357,P ＜ 0.05),the cell viabilities of dCK-WT,dCK-S74A and dCK-S74E showed no changes (P ＞ 0.05).The total mortalities of dCK-WT and dCK-S74E decreased significantly as compared with dCK-Vector (x2 =3.857-3.971,P ＜ 0.05),but no changes in dCK-S74A cells (P ＞0.05).The apoptosis rates in dCK-S74A,dCK-Vector and control group were up-regulated after irradiation (t =-4.531,-3.688 and-7.076,P ＜ 0.05),and the irradiation-induced apoptosis was reversed in dCK-WT and dCK-S74E (66％ and 68％ of the increase level in dCK-Vector group).The autophagy in dCK-WT and dCK-S74E increased by 22％ and 26％ (t =-9.051 and-8.411,P ＜0.01),but no changes were observed in dCK-S74A,dCK-Vector and control groups (P ＞ 0.05).Conclusions The dCK-WT and dCK-S74E could reverse the irradiation-induced apoptosis,increase the autophagy occurence,and decrease the total mortality,indicating that the phosphorylation of dCK at Ser-74 sites is related to the radiosensitivity of MCF-7 cells.

OBJECTIVETo detect the expression of the peripheral blood P2X5 receptor at various ambient temperatures, and to explore its relationship with deficiency-cold syndrome and deficiency-heat syndrome.

METHODSSubjects were selected by questionnaire and expert diagnosis, and assigned to the normal control group, the deficiency-cold syndrome group, and the deficiency-heat syndrome group, 20 in each group. 5 mL venous blood was collected at room temperature (25 °C) and cold temperature (-4-5 °C) respectively. Then the expression of P2X5 receptor was relatively quantified by real-time fluorescence quantitative PCR, and compared at room temperature and cold temperature respectively.

RESULTSThe expression of P2X5 receptor in deficiency-cold syndrome and deficiency-heat syndrome groups was lower than that in the normal control group at room temperature (P < 0.05). It decreased more at cold temperature in the deficiency-cold syndrome group than in the normal control group (P < 0.01) as well as in the deficiency-heat syndrome group (P < 0.05). The expression of P2X5 receptor showed no difference in all groups at two different temperatures (P > 0.05).

CONCLUSIONSThe expression of P2X5 receptor was different in different syndrome groups at various ambient temperatures. Ambient temperatures had insignificant effect on the expression of P2X5 receptor of the population with the same syndrome.

Cold Temperature ; Hot Temperature ; Humans ; Medicine, Chinese Traditional ; Receptors, Purinergic P2X5 ; metabolism ; Syndrome

OBJECTIVETo explore activity laws of mitochondrial complex II in patients of deficiency-cold syndrome (DCS) and deficiency-heat syndrome (DHS) under various ambient temperatures.

METHODSSubjects were recruited by questionnaire and expert diagnosis from grade 1 - 3 undergraduates at Henan College of Traditional Chinese Medicine in November 2012, and assigned to a normal control group, the DCS group, and the DHS group, 20 in each group. Their venous blood samples were collected at two different temperature conditions. Activities of mitochondrial complex II were measured by spectrophotometry.

RESULTS(1) Comparison of mitochondrial complex It under various ambient temperatures: Compared with room temperature in the same group, activity values were all increased in the normal control group at cold temperature with significant difference (P <0.05), but there was no significant difference in the DCS group and the DHS group (P >0. 05). Compared with the normal control group, activity values of complex H were reduced in the DCS group at cold and room temperatures with significant difference (P <0.05). Compared with the DCS group, activity values of complex It were increased in the DHS group with significant difference (P <0. 05). (2) Changes of adjustment rates: Compared with room temperature, the adjustment rate all rose at cold temperature in the normal control group and the DHS group with significant difference (P <0.05), but with no significant difference found in the DCS group (P >0. 05). Compared with the normal control group at the same temperature, the adjustment rate in the DHS group and the DCS group was all reduced at cold and room temperatures with significant difference (P <0. 05). There were no significant difference in the adjustment rate between the DHS group and the DCS group (P > 0. 05).

CONCLUSIONSEnvironment temperature can affect the activity of mitochondrial complex II with different influence degrees on different syndrome types of people, but its change trend are basically identical.

Cold Temperature ; Electron Transport Complex II ; metabolism ; Hot Temperature ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Temperature

Objective To investigate the effect of prognostic factors and nursing care in of acute myocardial infarction(AMI)patients performed emergency percutaneous coronary intervention supported by IABP.Methods Twenty seven AMI patients treated with emergency PCI,supporting by intra-aortic balloon pump(IABP).We analyzed retrospectively elinical characters and situation of interventional treatment of the patients.Results Among 27 cases.there were 12 cases who were recovered,4 suffered congestive heart failure repeatedly.and ll died during the hospitalization.There was no difference in age,sex,the history of hypertension,diabetes and smoking,the site of infarction and ejection fraction of left ventficular.The prognosis was different in the patients with grade of Killip in cardiac function and no flow post PCI(P<0.05).Conclusions The grade of Killip and no flow post PCI were the effective factors for prognosis in Acute Myocardial patients with emergency PCI.

Xylitol has attracted much attention because of its many applications tn the food,medicine and chemical industries. However the use has been limited by its high price. This coast is a result of the extensive purification steps needed for the preparation of a pure xylose solution,which is essential for the chemical process. The fermentative process of xylitol is an interesting alternative to conventional chemical process,since it does not require initial xylose purification. The present review describes the advantage of xylitol production by fermentation, xylitol-producting microorganisms, metabolic pathway of xylose in yeasts, detoxification of hemicellulose hydrolysates and fermentative conditions affecting xylitol production.

OBJECTIVETo investigate the effect of cytomegalovirus infection on expression of matrix metalloproteinase-2 in human endothelial cells.

METHODSHuman umbilical vein endothelial cells were cultured in vitro. Cells between 3-6 passages were infected with cytomegalovirus for different time. Expression of matrix metalloproteinase-2 mRNA was analyzed by reverse transcription-polymerase chain reaction, matrix metalloproteinase-2 activity was detected by gel zymography.

RESULTSExpression of matrix metalloproteinase-2 mRNA and its activity 6 hours after infection was almost equal to control, and was greatly enhanced 12 and 24 hours after infection.

CONCLUSIONCytomegalovirus infection up-regulates expression of matrix metalloproteinase-2 in human endothelial cells. It might be one of the mechanisms that cytomegalovirus is involved in atherosclerosis.

Cells, Cultured ; Cytomegalovirus ; physiology ; Endothelial Cells ; cytology ; metabolism ; virology ; Gene Expression Regulation, Enzymologic ; Host-Pathogen Interactions ; Humans ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Umbilical Veins ; cytology

The effects of the concentration of sulfuric acid and the ratio of liquid to solid on xylose yield from sugar cane bagasse in its hemicellulose hydrolysis process were studied with the Quadratic Rotary Combination Design. Regression analysis showed that there was a marked regression relationship between the two factors and xylose yield. As the result of optimizing the hydrolysis conditions by regression equation, xylose yield of 24 g/100 g sugar cane bagasse was obtained when sulfuric acid concentration was 2.4 g/L and liquid to solid ratio was 6.2 under the conditions of stream pressure of 2.5 x 10(4) Pa and hydrolysis time of 2.5 h. The macroporous resin adsorption was proved to be a good method to reduce the concentration of yeast cell growth inhibitor in sugar cane bagasse hemicellulose hydrolysate and to enhance the hydrolysate fermentability. The hydrolysate treated with macroporous resin adsorption under pH2 was used as the substrate for xylitol production by a xylitol-producting yeast, Candida tropicalis AS2.1776. At an initial xylose concentration of 200 g/L, all xylose was consumed within 110 h with a xylitol production rate of 1.15 g/L.h, and a xylitol yield of 0.64 g/g xylose.
Candida tropicalis ; metabolism ; Cellulose ; metabolism ; Fermentation ; Hydrogen-Ion Concentration ; Hydrolysis ; Polysaccharides ; metabolism ; Regression Analysis ; Saccharum ; metabolism ; Xylitol ; biosynthesis

OBJECTIVETo study the changes in the proliferation and synthetic function of human periodontal ligament fibroblasts (HPDLF) in response to dynamic mechanical strains of different modes, magnitudes and durations.

METHODSUsing a 4-point bending system, the effect of dynamic mechanical strains of different modes, magnitudes and durations on the proliferation of HPDLF was investigated by analyzing the cell cycle changes with flow cytometry (FCM), and the total protein level and the activity of alkaline phosphatase (ALP) in HPDLF were assayed by quantitative analysis.

RESULTSThe percentage of G(0)/G(1) cell decreased, S phase cells increased, and the proliferation index (PI), total protein level and activity of ALP were augmented significantly in response to dynamic mechanical micro-strains. These changes showed close correlations to the magnitude and duration of the strain. The mode of strain caused significant changes in G(0)/G(1), S, and G(2)/M phase cell percentages as well as the PI, total protein level and ALP activity of the cells. In the gradient strain group, the cell proliferation activity, total protein level and ALP activity were obviously higher than those in 1000 and 4000 microstrain groups.

CONCLUSIONThe changes in the proliferation and synthetic function of HPDLF are closely correlated to the mode, magnitude and duration of the strains.

Alkaline Phosphatase ; metabolism ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Flow Cytometry ; Humans ; Periodontal Ligament ; cytology ; Proteins ; analysis ; Stress, Mechanical

BACKGROUNDThe level of basic fibroblast growth factor (bFGF) increases rapidly after cerebral ischemia. However, the molecular mechanisms for the effects of bFGF on cerebral microvascular endothelial cells (cMVECs) have not yet been fully elucidated. In this study, a murine cMVEC line, bEnd.3, was employed to study the effects of bFGF on cyclooxygenase (COX) expression and its downstream effects in cMVECs.

METHODSAfter treatment with bFGF, RT-PCR and Western blotting analyses were carried out to evaluate the changes in COX-2 mRNA and protein expression, respectively. MTT assays were performed to measure cell proliferation. The prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA).

RESULTSCOX-2 mRNA and protein expressions in bEnd.3 cells were induced by bFGF in time- and dose-dependent manners. The bFGF-induced COX-2 upregulation led to enhanced PGE2 production by bEnd.3 cells, and this effect was abolished by the selective COX-2 inhibitor NS-398. bFGF also increased VEGF production by bEnd.3 cells, and this effect was blocked by NS-398 and the EP1/2 (PGE2 receptors) antagonist AH6809. Furthermore, exogenous PGE2 increased VEGF production in bEnd.3 cells, and AH6809 blocked this effect.

CONCLUSIONbFGF increases VEGF production in an autocrine manner by increasing COX-2-generated PGE2 in cMVECs and subsequently stimulates MVEC proliferation and angiogenesis.

Blotting, Western ; Cell Line ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; genetics ; metabolism ; physiology ; Dinoprostone ; metabolism ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Receptors, Prostaglandin E ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; metabolism ; Xanthones ; pharmacology

OBJECTIVETo establish a LC-MS/MS method for determining the concentration of tanshinone IIA, salvianolic acid B and paeoniflorin of refined coronary cataplasm in rabbit plasma, in order to determine the concentration of the three main ingredients in blood after transdermal administration and calculate their pharmacokinetic parameters.

METHODRabbits were given refined coronary cataplasm on the basis of 15 g x kg(-1) by transdermal administration to detect the plasma concentration of the three main ingredients using LC-MS/MS. Winnonlin software was used to calculate their major pharmacokinetic parameters.

RESULTTanshinone IIA, salvianolic acid B and paeoniflorin showed good linearity (r>0.999) at 1-100, 50-1 000, 10-1 000 microg x L(-1) respectively in plasma, with average recovery rate of 96.57%, 91.90%, 95.93%, respectively. The RSD within day were less than 15%. After transdermal administration of refined coronary cataplasm in rabbits, the main pharmacokinetic parameters of tanshinone IIA, salvianolic acid B or paeoniflorin were as follows: Cmax (20.85 +/- 12.68), (636.25 +/- 386.91), (787.80 +/- 395.64) microg x L(-1); Tmax (0.49 +/- 0.28), (0.44 +/- 0.27), (0.46 +/- 0.30) h.

CONCLUSIONThe LC-MS/MS method is highly selective and sensitive to determine the concentration of samples in rabbit plasma. The pharmacokinetic characteristics of tanshinone IIA, salvianolic acid B and paeoniflorin are suitable to assess the percutaneous absorption of refined coronary cataplasm.

Animals ; Benzoates ; administration & dosage ; pharmacokinetics ; Benzofurans ; administration & dosage ; pharmacokinetics ; Bridged-Ring Compounds ; administration & dosage ; pharmacokinetics ; Chromatography, Liquid ; methods ; Coronary Disease ; drug therapy ; Diterpenes, Abietane ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; pharmacokinetics ; Humans ; Male ; Monoterpenes ; Rabbits ; Tandem Mass Spectrometry ; methods