Preparation of glycine liposomes and it's effect on cardiomyocyte injury induced by hypoxia/reoxygenation

AIM: To prepare glycine liposome microparticle and observe the effect of glycine liposomes on cardiomyocyte injury induced by hypoxia/reoxygenation. METHODS: （1） Reverse-phase evaporation method was used to produce glycine liposomes, the effects of different organic solvents: aether, chloroform and two mixtures of aether/chloroform on entrapment efficiency were evaluated, transmission electron microscope was used to detect the particle diameter of glycine liposomes. （2） A cardiomyocyte injury model was established by using hypoxia/reoxygenation, the activities of lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase isoenzyme (CK-MB) of each group were detected. RESULTS: The entrapment efficiency of glycine liposomes prepared with the mixtures of aether/chloroform is highest compared with other organic solvents （64.8％, P<0.01）; all of glycine, glycine liposomes and blank liposomes inhibited the release of LDH, CK and CK-MB induced by hypoxia/reoxygenation in cultured cardiomyocytes, and the inhibition by glycine liposomes was the most obvious （P<0.05，P<0.01）. CONCLUSIONS: The mixtures of aether/chloroform can be used to prepare glycine liposomes, which can get higher entrapment efficiency; glycine liposomes protects cultured cardiomyocytes against hypoxia/reoxygenation-induced injury, and the protective effect of glycine liposomes is better than that of glycine.     

Effects of activated state of T cells from human peripheral blood on absorption of photosensitizer hematoporphyrin monomerthyl ether

AIM: To investigate the characterization of absorption of hematoporphyrin monomerthyl ether (HMME), a domestic new generation photosensitizer product, by activated T cells from human peripheral blood. METHODS: Evaluation was performed by flow cytometry on the effects of incubating concentration and time of HMME on absorption by activated T cells. Lymphocytes were separated from human peripheral blood by density gradient centrifugation with Ficoll and T cells were activated with polyclonal stimulators PHA and PDB+Ion. To analyze the effects of HMME incubating doses on the absorption of activated T cells, the cultural lymphocytes were incubated with a serial doses of HMME for 1 h and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. To test the impact of HMME incubating time on the absorption of activated T cells, the cultural lymphocytes were incubated with HMME for various times and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. RESULTS: The HMME absorption-dose curve and absorption-time curve were shifted to right and up in the activated T cells as compared to resting T cells. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the doses between 5 mg/L to 20 mg/L. HMME absorptions of either activated T cells or resting T cells underwent a gradual increase with the incubation-time in HMME at concentration of 10 mg/L. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the incubation-time between 15 to 60 min. CONCLUSION: The differences of HMME absorption between activated T cells and resting T cells depend on the incubation times and doses of HMME. HMME absorption of activated T cells are significantly larger than that of resting T cells in certain incubation-times and doses. These results suggest that incubation time and dose associated with HMME-PDT therapeutic windows will be created for selective deletion of activated T cells.     

Exogenous carbon monoxide protects against the lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To study the mechanism of protective effect of exogenous carbon monoxide (CO) in the lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats. METHODS: Thirty-two SD rats were randomly divided into 4 groups: control, control+CO, IR and IR+CO. A rat model of ischemia in hind limbs and the reperfusion lung injury was made. The rats in IR+CO and control+CO groups were exposed to air containing 2.5×10-8 CO for 1 h before reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The lung tissue structure, polymorphonuclear leukocyte (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content and the animal survival rate were observed. The carboxyhemoglobin (COHb) levels in artery blood were detected with CO-oximeter and the expression of intercellular adhesion molecule-1 (ICAM-1) in the lung was detected by Western blotting. RESULTS: Compared to control, the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly increased in IR group. Compared with the IR group, the blood COHb level was significantly increased and the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly decreased in IR+CO group. CONCLUSION: These data suggest that exogenous CO attenuate limb IR-induced lung injury by down-regulatiny ICAM-1 expression and suppressing PMN sequestration in the lung following limb IR in rats.     

Influence of ovariectomy and estrogen replacement on gene expression profile of myocardium in rats

AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.     

Biological characters of mesenchymal stem cells of human umbilical cord blood

AIM：To study the isolation,expansion and purification of mesenchymal stem cells (MSCs) from human umbilical cord blood (UCB),and investigate some biological identities of MSCs.METHODS：(1) MSCs of UCB,adult bone marrow (BM) and fetus BM were isolated by centrifugation with Ficoll,and the different kinds of MSCs were observed everyday.(2) Surface markers of MSCs were identified by flow cytometry.(3) The level of HGFs (TPO,SCF,FLT-3L,IL-6) secreted by different sources of MSCs was checked by ELISA method.RESULTS：(1) No difference in morphology of the colonies between UCB MSCs and BM MSCs was observed.However,the mononuclear cells needed in culture of UCB MSCs was about 3 times more than that in culture of BM MSCs.The times of UCB MSCs colony formation and confluencing were longer than that of BM in primary culture.(2) After passaged,there was no significant difference in the proliferation rates of 3 kinds of MSCs.Only 4 of 15 UCB samples contained a homogeneous population of MSCs.(3) UCB MSCs shared the same markers with BM MSCs.Neither hematopoietic marker nor immunologic recognition antigens were expressed.(4) The level of hematopoietic growth factors (HGFs) secreted by 3 kinds of MSCs was similar.CONCLUSIONS：(1) MSCs were isolated from UCB,but the amount of MSCs in UCB was smaller than that in BM,and just seldom samples of UCB contained homogeneous MSCs.(2) MSCs from UCB and BM shared the same biological characteristics,such as proliferation ability,surface markers,immunophenotypes and HGFs secretion.     

二氯喹啉酸诱导烟草过氧化物酶 活性的动态分析

为了验证不同剂量二氯喹啉酸致烟草畸形生长与诱导植物体内保护酶系中的过氧化物酶活性间的关系,采用盆栽试验的方法,通过添加二氯喹啉酸,模拟田间二氯喹啉酸在土壤中的不同残留量,造成烟草不同程度的畸形生长,在不同时间内,测定烟叶中过氧化物酶(POD)活性的变化。结果表明,处理组烟叶中POD酶的活性明显高于未用药的对照烟叶,随着处理浓度的升高和时间的延长,POD酶的活性有上升趋势,当处理28天后,各处理的二氯喹啉酸浓度为1.04×10-3、2.08×10-3、4.17×10-3、8.33×10-3和1.67×10-2mg/kg,对照和各处理的过氧化物酶活力分别为17.47、29.61、23.73、30.33、41.00、57.73OD·g-1·min-1,烟叶中过氧化物酶活性较对照分别提高69.47%、35.80%、73.61%、134.69%和230.45%。POD同工酶的检测表明,处理过的烟叶酶带增多、颜色加重、酶活增强。     

不同抗性玉米品种接种甘蔗花叶病毒(SCMV)后4种防御酶活性变化研究

 植物在长期进化过程中,不断抵御外部伤害,包括有害生物的入侵,其体内逐渐形成了一套完整的保护机制。     

植物提取物、碳酸钠和碳酸氢钠室内对杨树溃疡病菌生长的抑制作用

 采用菌丝生长速率法用金银花的花、黄花蒿的地上部分、蓝桉果实和黄柏果实的乙醇提取物及其不同极性的组分对杨树溃疡病菌菌丝生长的抑制作用进行了测定。以黄柏果实的抗菌活性最强,其次是金银花的花。抗菌活性成分主要存在于金银花的正丁醇组分、黄花蒿的石油醚组分和乙酸乙酯组分、蓝桉的水部分、黄柏的正丁醇组分。当培养基中碳酸钠和碳酸氢钠的浓度分别为8g/L时(对应的pH值分别为10.24和7.71),菌丝生长抑制率分别为100.00%和79.68%。如果用1mol/L氢氧化钠溶液调节培养基的pH值至10.00,菌丝生长抑制率为40.58%。说明碳酸钠或碳酸氢钠对菌丝生长的抑制作用,一方面是由于改变了培养基的pH值,另一方面可能是由于碳酸根和碳酸氢根离子抑制了菌丝的生长。     

新化合物HNPC-B2055的室内除草活性研究

HNPC-B2055是一种具有极高苗后除草活性的新化合物,在室内进行了该化合物对靶标杂草的IC50(或IC90)值测定、杀草谱测定、对作物安全性测定、吸收及传导性测定、土壤持效性测定以及对后茬作物的安全性测定。