Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.

Objective To probe the effects of hypothermia and St Thomas Hospital Ⅱ cardioplegia on immature myocardium.Methods To observe the change of hemodynamics,myocardial enzyme in the coronary effluent and myocardial biochemistry of the perfused immature rabbit heart in vitro after two or four hours ischemia at 14 degrees centigrade.Results There were no significant differences in post ischemic hemodynamics,myocardial enzyme in the coronary effluent and myocardial biochemical parameters of the perfused immature heart The myocardial protective effect provided by St.Thomas Hospital Ⅱ cardioplegia was worsen than that of hypothermia alone,marked by the elevated myocardial enzyme leakage and the decreased hemodynamics.Single dose perfusion was better than multi dose perfusion by characteristics of decreased enzyme leakege and good post ischemic hemodynamics.Conclusions Hypothermia alone can provide immature heart with satisfactory myocardial protection.St.Thomas Hospital Ⅱ cardioplegia can not afford good myocardial protection to immature heart and enhance the myocardial protective effect provided by hypothermia.The myocardial protection effect is better provided by single dose perfusion than by multi dose perfusion

The current workflow in home-based rehabilitation for stroke patients were modified through focus group discussion,brainstorming and literature review.The mapping of three-tier rehabilitation network,the defined function of home-based rehabilitation and its integration with other rehabilitation services were made by literature review.The improved version of home-based rehabilitation flow chart specified the contents in the guideline Three-tier rehabilitation network of stroke rehabilitation in China.The chart provides a reference for the implementation of home-based rehabilitation in community health service centers with the family doctor system.

OBJECTIVETo compare the photosynthetic characteristics difference of different ploidy Rhodiola sachalinensis germplasm and provide the scientific basis for their cultivation.

METHODLI-6400/XT photosynthesis system was used to measure leaf light response curve and CO2 response curve of diploid and autotetraploid. Biomass, leaf area, stomatal characteristics and chlorophyll content differences were compared in the study.

RESULTStomata of the two germplasms were open during daytime obviously, and stomata conductance responded to the changes of light intensity and CO2 concentration which was not consistent with the characteristics of CAM (crassulacean acid metabolism) plants. Light compensation point of autotetraploid was significantly lower than that of the diploid, and light saturation points of both germplam were close, and their light saturation points were near 500 micromol x m(-2) x s(-1). Quantum efficiency of autotetraploid was significantly higher than the diploid, and the net photosynthetic rate of autotetraploid significantly higher than the diploid when light intensity was higher than 500 micromol x m(-2) x s(-1). Stomata conductance, transpiration rate of autotetraploid was also significantly higher than that of diploid. Biomass, leaf area, stomata diameter and chlorophyll content of autotetraploid were much higher than that of diploid, while the stomata density of autotetraploid was less than diploid.

CONCLUSIONThe results above provide scientific basis for the cultivation of different ploidy Rh. sachalinensi germplasm.

Carbon Dioxide ; metabolism ; Photosynthesis ; physiology ; Ploidies ; Rhodiola ; metabolism

AIM:The objective of this study was to observe the effects of AP-1 element on endothelin-1(EDN1)gene transcription in homocysteine(Hcy)-induced human umbilical vein endothelial cells(HUVECs).METHODS:The redox level in Hcy-induced HUVECs was determined by using the fluorescent product DCF.The influences of Hcy on expressions of EDN1 mRNA and protein of c-jun/AP-1 in HUVECs were measured by the methods of RT-PCR and Western blotting,respectively.The EDN1 secretion level of HUVECs induced by Hcy was determined by enzymatic immunoassay.The transient transfection assay was performed and luciferase activity of transcriptional factor AP-1 reporter gene induced by Hcy was detected.RESULTS:The Hcy significantly increased EDN1 secretion,EDN1 mRNA expression and oxidative stress in HUVECs.Hcy remarkably induced the expression of pGL3-EDN1-AP-1(115/+135)luciferase reporter gene plasmid.However,basic expression of mutation of pGL3-EDN1-AP-1(-115/+135)luciferase reporter gene plasmid was downregulated markedly in HUVECs induced by Hcy.CONCLUSION:Redox-active and release of ROS are changed by Hcy.Activated AP-1 element plays an important role in EDN1gene transcription in HUVECs induced by Hcy.

ObjectiveTo investigate the effect of α-Zearalanol(α-ZAL) on lipometabolism and hemorheology in ovariectomized(OVX) hyperlipidemia rabbits.Methods44 adult virgin female rabbits were divided into 5 groups,group A: normal control;group B: sham+CHO;group C: OVX+CHO;group D: OVX+CHO+17βE_2;group E: OVX+CHO+α-ZAL.Cholesterol(CHO) was fed to rabbits for 12 weeks.Before and after feeding CHO,the serum lipid(TC,TG,LDL-C,HDL-C) were measured;Blood viscosity,plasma viscosity,aggregation index of RBC(AIRC) and fibrinogen were also assayed respectively.ResultsThe serum levels of TC,TG and LDL-C in group B and E were significantly decreased compared with those in group C(P<0.05);the level of blood viscosity,plasma viscosity and AIRC platelet aggregation rate in group D and E were also significantly decreased compared with those in group C(P<0.05).Conclusionα-ZAL can improve vascular function through the adjustment of lipometabolism and hemorheology.

AIM: To investigate the effect of 17? estradiol (17?E 2)on atherosclerosis and hemorrheology in ovariectomized (OVX) rabbits. METHODS: Thirty four female rabbits were divided into 4 groups, group A: normal control; group B: sham+CHO; group C: OVX+CHO; group D: OVX+CHO+17? E 2. Cholesterol(CHO) was fed to rabbits for 12 weeks, before and after feeding CHO, the serum lipid (TC, TG, HDL-C, LDL-C, ApoA1, ApoB) and area of aortic atherosclerotic plaques were measured. Blood viscosity, plasma viscosity (?p), aggregation index of RBC (AIRC) and fibrinogen were also determined, respectively. RESULTS: ① The ratio of area of aortic atherosclerotic plaque to total area of aortic intima was 0.02?0.003 (in group A), 0.42?0.15 (group B), 0.67?0.23 (group C), and 0.12?0 11 (group D), respectively. In group D, the ratio of aortic atherosclerotic plaque was markedly decreased compared with group C ( P

OBJECTIVETo acquire homozygous tetraploid germplasm of Rhodiola sachalinensis.

METHODPEG-mediated protoplast fusions were conducted using callus of Rh. sachalinensis as materials. Protoplast fusion products were embedded and cultured in low-density, low-melting-point agar and marked according to the protoplast size, and single-celled sister lines were established to acquire genetically homozygous tetraploid germplasm.

RESULTR(D) and R(M) of newborn daughter cells or protoplasm, metaphase cells or protoplasm were approximately in line with the formula R(D) = 0.793 7R(M). The change range in diameter of the diploid cells without fusion, two protoplasts fusion product were: 16.7 microm < or = R < 21.3 microm, 21.0 microm < or = R' < 26.8 microm respectively. There is an overlap between the two diameter ranges. The protoplast inoculation density of 1 x 10(4) cells x mL(-1) was appropriate when protoplasts were anchored by low-intensity, low-melting-point agar. Under the conditions of this density, plating efficiency was high and single cell origin of the sister lines microclones grew rapidly, and it was easy to mark the single cell microclones, and separate from each other to subculture. The chromosome counts results showed that chromosome numbers of diploid and tetraploid of single cell lines were 26 and 52, respectively. The result from flow cytometry assay showed that there is no presence of chimerism in single-cell regeneration plantlets.

CONCLUSIONThe results of this study provide a scientific basis for polyploid breeding of Rh. sachalinensis.

Cell Fusion ; methods ; Polyethylene Glycols ; pharmacology ; Polyploidy ; Protoplasts ; cytology ; drug effects ; Rhodiola ; cytology ; drug effects ; genetics

Objective To establish the chronic low flow myocardial hibernation animal model in pigs, and to assess the diagnostic value for myocardial hibernation by using various imaging methods. Methods A total of 13 miniswine (30-40 kg) were used. All animals underwent general anesthesia and orotracheal intubation while the animals were mechanically ventilated. Under sterile conditions, left ventriculography and coronary angiography were performed by introduction of catheter into the right femoral artery. Further, a left anterolateral thoracotomy was performed in the third intercostal space. The proximal LCX was dissected free to allow placement of an ameroid constrictor. More than 1 month later, left ventriculography and coronary angiography were performed again, followed by cine MRI at rest and during stress with low dose of dobutamine (5 ?g?kg -1 ?min -1 ), respectively. Traditional and/or breath hold cine MRI were used to evaluate regional left ventricular wall motion, corresponding to basal, midventricular and apical short axis tomograms. Regional wall motion score index (WMSI) was calculated. At the same time 99m Tc MIBI myocardial SPECT was performed at rest and during nitroglycerin administration, respectively. All animals were finally sacrificed for pathological examination. Triphenyl tetrazolium chloride (TTC) staining was used to assess the myocardial infarction. Electron microscopy was used to identify myocardial cellular changes characteristic of hibernating myocardium. Results Three pigs died during surgery or within two weeks after surgery. One pig died of anesthesia during SPECT examination, 1 pig suffered from aneurysm, and another one pig showed negative findings. The other 7 pigs were found with hypokinetic ( n =4) or akinetic ( n =3) myocardial regions related to stenosed LCX (70%-99%). Resting cine MRI demonstrated decreased regional motion of the lateral and posteroinferior walls (ischemic regions) of the left ventricle ( n =7), compared with the nonischemic anteroseptal regions; but the low dose dobutamine (5 ?g?kg -1 ?min -1 ) could recover those hypokinetic or akinetic myocardial regions, characteristic of hibernating myocardium. Resting 99m Tc MIBI myocardial SPECT ( n =6) showed a fixed perfusion defect on the corresponding ischemic areas, which became reversible on the nitrate augmented myocardial perfusion imaging. It also indicated myocardial viability presented at the ischemic areas. TTC staining revealed patchy infarction of the area at risk localized to the endocardial surface ( n =3), and no myocardial infarction ( n =4). Electron microscopy of sections from the hibernating regions revealed loss of contractile materials, increased numbers of small mitochondria, and glycogen accumulation within viable cardiomyocytes, which had been described as hallmarks of hibernating myocardium. Conclusion Chronic low flow myocardial hibernation can be reproduced in an animal model during progressive coronary stenosis caused by ameroid constrictor.

OBJECTIVETo evaluate the diagnostic value of dobutamine stress magnetic resonance imaging (MRI) for myocardial viability.

METHODSTen male miniswines underwent left ventriculography and coronary angiography, followed by stenosis of the left circumflex coronary artery (LCX) using ameroid constrictor. More than one month later, left ventriculography and coronary angiography were performed again, followed by cine-MRI at rest and during stress with incremental dose of dobutamine 5 - 20 micro g.kg(-1).min(-1). Traditional and/or breath-hold cine-MRI were used to evaluate regional left ventricular wall motion, corresponding to basal, midventricular and apical short-axis tomograms. Regional wall motion score index (WMSI) was calculated. The miniswines were finally sacrificed for pathological examination. Triphenyl tetrazolium chloride (TTC) delineated myocardial infarction. Microscopy was used to identify myocardial cellular changes.

RESULTSOne pig died, one pig suffered from aneurysm and another showed no negative findings. The other seven pigs were found with hypokinetic (n = 4) or akinetic (n = 3) myocardial regions related to stenosed LCX. Their mean WMSI at rest for the lateral and posteroinferior walls (ischemic regions) of the left ventricle was 2.27 +/- 0.32, as compared with 1.00 +/- 0.00 (P < 0.01) for the corresponding nonischemic anteroseptal regions. Further, the mean WMSI for the ischemic regions was 2.27 +/- 0.32 at rest compared with 1.40 +/- 0.39 (P < 0.01) at the dose of dobutamine 5 micro g.kg(-1).min(-1). However, the mean WMSI at the doses of dobutamine 10 and 20 micro g.kg(-1). min(-1) were 1.70 +/- 0.76 and 1.75 +/- 0.83, respectively, with no significant difference as compared with the mean WSCI at rest (P > 0.05). The pathologic examination showed viable myocardium at the ischemic regions.

CONCLUSIONLow-dose dobutamine (5 micro g.kg(-1).min(-1)) recovers hypokinetic or akinetic myocardial regions, and dobutamine stress MRI can be used to detect myocardial viability.

Animals ; Dobutamine ; Magnetic Resonance Imaging, Cine ; methods ; Male ; Swine ; Swine, Miniature ; Ventricular Function, Left