Aim To investigate the effects and mechanisms of DMA on cardiac function in cardiomegaly rat isolated hearts.Methods With Langendorff perfusion,the changes in the indicators reflecting cardiac function were observed in cardiomegaly rat isolated hearts.Its probable mechanism was explored with calcium channel blocker and sodium calcium exchange(NCX) blockers.Results DMA(0.5～2 ?mol?L~(-1))enchanced cardiac function and exerted positive effects on LVSP-LVDP,+dp/dt_(max)and-dp/dt_(max) in Langendorff-perfused cardiomegaly rat isolated hearts.The effects of DMA on cardiac function in cardiomegaly rat isolated hearts weren′t blocked by nicardipine-one of calcium channel blockers and were blocked by NiCl_2——one of sodium calcium exchange(NCX) blockers.Conclusion DMA can enhance systolic and diastolic function in cardiomegaly rat isolated hearts.By agitating NCX,DMA enhances cardiac function in cardiomegaly rat isolated hearts,independent of L-calcium channel.

Objective To evaluate the heating ability of thermoseeds in alternating magnetic field and its therapeutic effects on tumor bearing rats with simultaneously detected immune function. Methods To monitor the temperature increase of thermoseeds in vitro,of which the Curie point was 57 ℃ or 70 ℃.Forty Wistar rats implanted with Walker-256 cells were randomly divided into five groups: C group(control group,10 rats),M group(magnetic control group,5 rats),T group(thermoseeds control group,5 rats) and two heating treatment groups(H1 group,10 rats and H2 group,10 rats).In the H1 group,two thermoseeds with a Curie point at 57 ℃ were implanted into the tumor tissues in each rat with a heating time of 30 minutes,while in the H2 group,two thermoseeds with a Curie point at 70 ℃ were used with a heating time of 6 minutes.Five rats from each group were killed 9 days after the heating therapy to evaluate the volume and weight of the tumor tissues.The peripheral blood T-cells were counted in the rest rats in the H1,H2,and C groups. Results Magnetic inductive heating of the thermoseeds in vitro can reach the Curie point.The median tumor volume in the H2 group was 0.50 cm3(0.00-26.54),which was significantly lower than that in the M group(36.18 cm3,0.96-39.90,Z=2.21,P=0.032).And the weight of the tumors in the H1 and H2 groups was significantly lower than that in the M group [0.96 g(0.00-21.18) in H1 and 0.41 g(0.00-23.40) in H2 vs 31.45 g(1.09-36.09) in M group,Z=2.21 and P=0.032 for both].In the peripheral blood,the percentage of CD4+ T-cells was(22.39?5.27)% in H1 group,and(24.76?5.19)% in H2 group,which were significantly higher than that in C group [(12.07?4.45)%,P=0.01 and 0.003 respectively];and the percentage of CD8+ T-cells in the H2 group was significantly higher than that in the C group [(19.58?4.63)% vs(12.72?3.96)%,P=0.04].Conclusions Thermoseed had a good heating ability in alternating magnetic field and its inductive heating can inhibit tumor growth in Wistar rats and improve the immune function of the rats.

Objective To study the in vitro heating ability of Ni-Cu thermoseeds and their effect on the rabbit liver cells and tissues. Methods The temperature of rabbit liver tissues were monitored under an alternating magnetic field.MTT assay was used to evaluate the in vitro cytotoxicity of the extra-liquid of the Ni-Cu thermoseeds;Hemolytic test was carried out to estimate its blood toxicity;and muscular implantation test was employed to determine the levels of its tissue toxicity.Results The thermoseeds used in this experiment showed a high heating ability in alternating magnetic field in vitro.MTT assay showed that the toxicity of the material on mouse fibroblast(L-929) cell lines was 1 degree,which means non-cytotoxic.Hemolytic test revealed a hemolysis rate(HR) of 3.25%(less than 5%),showing that the thermoseeds had no hemolytic reaction.Muscular implantation test showed different levels of inflammatory reaction in the muscle tissues.Conclusion Thermoseeds induced heating in alternating magnetic field can achieve an appropriate temperature,and the gilded thermoseeds have a high biocompatibility with 1 degree cytotoxicity without leading to hemolytic reaction.

Objective To observe the effect of retrograde sinus coronary perfusion on the function of myocardial cell membrane following retrograde sinus coronary perfusion Methods Thirty two isolated rabbit hearts were randomized into 4 groups: Group A, hearts were kept beating by retrograde perfusion with normothermic oxygenated blood; Group B, hearts received retrograde coronary sinus perfusion with warm blood cardioplegia; Group C, hearts received retrograde coronary sinus perfusion with cold blood cardioplegia; Group D, hearts received retrograde coronary sinus perfusion with cold crystalloid cardioplegia After 60 min retrograde perfusion, the hearts were switched on Langendorff Neely antegrade perfused working heart model for other 60 min Free calcium in myocardial cell, myocardial membrane MDA, ATPase and myocardial cell membrane fluidity were measured Results After retrograde coronary sinus perfusion, activity of ATPase and fluidity of the myocardial cell membrane in group A were higher than those in groups B, C and D The contents of myocardial cell membrane MDA and calcium in myocardial cell following retrograde perfusion were much lower in group A than those in groups B and C There was no remarkable difference between group C and group D Conclusion Retrograde perfusion of beating heart is superior to those perfused with warm blood cardioplegia in reducing free radical production, ameliorating paradoxical calcium accumulation in myocardial cell and maintaining activity of ATPase and fluidity of the myocardial cell membrane Warm blood cardioplegia is superior to cold blood cardioplegia

Aim To study the inotropic action of DMA in normal rat isolated hearts and papillary muscle and search for its primary mechanism.Methods With Lansendorff-perfusion and isolated papillary muscle perfusion,cardiac performance and the systolic function of papillary muscle were measured to estimate the inotropic action of DMA;L-calcium channel blocker and sodium calcium exchanger(NCX)inhibitor were used to search for its primary mechanism.Results ①(1~20)?mol?L-1 DMA exerted a positive inotropic action in normal rat isolated hearts,that is in Langendorff-perfused hearts,DMA enhanced cardiac performance,i.e.LVSP-LVDP,+dp/dtmax,-dp/dtmax significantly(P

Objective A new breed of licorice seeds(Wuxin No.1) was selected from wild licorice,Glycyrrhiza uralensis.Methods Its germination rate,content of soluble protein,and peroxidase(POD) activities were investigated and compared with the wild licorice seeds(control group).The leaves of Wuxin No.1 were collected and its genetic polymorphism was analyzed by inter-simple sequence repeat technique(ISSR).Results The results suggested that the seed vitality in Wuxin No.1 was higher than that in the control group.The change of soluble protein and POD activity also demonstrated that the seeds in Wuxin No.1 have the higher vigor.ISSR Analysis showed that among 22 random primers used in this experiment,six primers generated different DNA band types,which meant that there was genetic polymorphism in Wuxin No.1.Conclusion All these changes indicate that Wuxin No.1 is a prospective domestication species of licorice and may be cultivated widely in the future.

OBJECTIVETo substantiate the effects of spaceflight on the glycyrrhizic acid-related gene mutation in Glycyrrhiza uralensis.

METHODLicorice (G. uralensis) seeds were carried by a recoverable satellite for 18 days (the average radiation dose in the flight recovery module was 0. 102 m x d(-1), the orbit semidiameter 350 km, gravity 10(-6)). After returned to the earth, the satellite-flown seeds and the un-flown seeds (ground control) were planted in the fields of experimental farm. The leaves of each group were used for studying the effects of space flight on the glycyrrhizic acid-related gene mutation including ITS sequence and beta-amyrine synthase gene.

RESULTThe ITS sequence of glycyrrhizic acid related gene showed no changes after spaceflight. While beta-amyrine synthase gene had some different points after spaceflight and the different points could get the expression.

CONCLUSIONThe results indicated that spaceflight induce genetic variation in G. uralensis and spaceflight could also have effects on glycyrrhizic acid-related gene mutation in G. uralensis. It may need to further research how the spaceflight induced the mutation of the glycyrrhizic acid related gene. The results suggested that recoverable satellite-flown condition could bring inheritable mutagenic effects on G. uralensis seeds and maybe used as a tool for accelerating the progress in G. uralensis breeding.

Extraterrestrial Environment ; Glycyrrhiza uralensis ; genetics ; metabolism ; Glycyrrhizic Acid ; metabolism ; Mutation ; Plant Proteins ; genetics ; metabolism ; Space Flight

Space breeding in medicinal plants is special characteristics in China. Compared with other plants, in spite of a relatively small number, Medicinal plants have more obvious characteristics and advantages. Research on medicinal plants has also been carried into all aspects, such as biological traits, physiology and biochemistry, genomics, as well as differences in chemical composition, and chemical composition analysis is also involved. However, compared with other plants, especially crops and vegetables, biological research is an obvious deficiency, that is mainly reflected in the insufficient genetics and breeding researches, the stability of genetic traits from generation to generation were not followed up and in-depth study in breeding areas was not carried out. If medicinal plants resources from space with the genetic stability good quality were selected, it would address the problem of lack of resources and ease the pressure on wild resources of medicinal plants. It would at the same time play an important role in promoting the development of medicinal botany space breeding and the implementation of modernization of traditional Chinese medicine.
Breeding ; methods ; Plants, Medicinal ; growth & development ; physiology ; Space Flight ; Weightlessness

OBJECTIVETo investigate the space environment on the role of licorice mutagenesis analysis of proteins.

METHODLicorice (Glycyrrhiza uralensis) seeds were carried by a recoverable satellite for 18 days (the average radiation dose in the flight recovery module was 0.102 m x d(-1), the orbit semidiameter 350 km, gravity 10(-6)). After return, The satellite-flown seeds and the unflown seeds (ground control) were planted in the fields of experimental farm. The leaves of each group were used for studying the effects of space flight on CAT, SOD activity, the protein content and electrophoresis.

RESULTAfter the space flight, CAT, SOD activity of licorice increased in varying degrees, the difference was significant (P<0.05), two types of enzyme activity of sample from Ordos were higher than that from Hangjinqi. The protein content of licorice increased in a certain extent, the difference was significant (P<0.05), while protein electrophoresis also showed differences, weak new bands appeared.

CONCLUSIONThese results indicated that spaceflight has effect on protein of licorice, these changes may be used as a tool for accelerating the progress in G. uralensis breeding.

Chloramphenicol O-Acetyltransferase ; analysis ; metabolism ; Electrophoresis ; Extraterrestrial Environment ; Glycyrrhiza uralensis ; chemistry ; enzymology ; Plant Proteins ; analysis ; metabolism ; Spacecraft ; Superoxide Dismutase ; analysis ; metabolism

BACKGROUNDTo establish a sandwich ELISA method for detection of reporter chloramphenicol acetyltransferase (CAT) gene.

METHODSThe full length sequence of CAT gene was amplified with PCR using plasmid pBLCAT6 as template, and inserted into the prokaryotic expression plasmid Pgex-2T. The purified fusion protein was emulsified with complete or incomplete Freund adjuvant and injected subcutaneously into rabbits. The antibody was labeled with biotin, and a sandwich ELISA technique with biotin streptavidin amplify system was established. Several CAT reporter plasmids containing different HPV 16 LCR sequences were generated and transfected transiently to monolayer cells in vitro. The cytoplasm proteins were extracted and the expressions of CAT were evaluated with the newly established ELISA assay.

RESULTSSDS-PAGE displayed that the molecular weight of the expressed fusion protein was about 54,000. The prepared antiserum was able to recognize the CAT protein expressed by mammalian cells or prokaryote cells. Under the control of different promoters and their regulate sequences,two to eight folds CAT expression increased were evaluated in transiently transfected mammalian cells by the newly established sandwich ELISA method.

CONCLUSIONSThe established method could sensitively reflect the activities of the upstream promoters, as well as the influence of exchanges of nucleotides within the regulate region on the promoter activities. Therefore, it proposes a convenient assay for the studies using CAT as the reporter gene.

Animals ; Antibodies ; analysis ; Cells, Cultured ; Chloramphenicol O-Acetyltransferase ; analysis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Genes, Reporter ; Male ; Papillomaviridae ; genetics ; Plasmids ; genetics ; Rabbits ; Recombinant Fusion Proteins ; analysis ; biosynthesis ; immunology