Effect of endogenous H2S on pulmonary hypertension during acute lung injury induced by LPS and its interaction with NO

AIM: To investigate the effect of H2S on pulmonary artery hypertension during acute lung injury induced by LPS and the interaction between the systems of hydrogen sulfide (H2S)/cystathionine-β-lyase (CSE) and nitric oxide (NO)/nitric oxide synthase (NOS) in this process. METHODS: Seventy-two adult male rats were randomly divided into four groups: control group, LPS group, LPS+L-NAME group and LPS+propargylglycine (PPG) group. Mean pulmonary artery pressure (mPAP) of each rat was examined at 2 h, 4 h, 6 h and 8 h after treatment. H2S and NO contents in plasma, NO content, iNOS, cNOS and CSE activity in lung were measured at 4 h or 8 h after treatment, respectively. Expression of iNOS in lung tissue was also detected by immunohistochemistry technique, and the injury of lung was evaluated with morphological changes under microscope. RESULTS: LPS could induce severe lung injury, and mPAP, NO content, iNOS activity and its protein expression in LPS group significantly increased, but cNOS activity, H2S content and CSE activity decreased compared with those of control group. Administration of L-NAME before LPS could attenuate the changes induced by LPS. Pre-administration of PPG, a CSE inhibitor, exacerbated the injury by LPS, but there was no prominent variation in cNOS activity. CONCLUSION: Reduced endogenous H2S could increase pulmonary artery hypertension during acute lung injury induced by LPS. There is a negative effect between H2S/CSE system and NO/NOS system in this process.     

Effect of mitochondrial ATP sensitive potassium channels opening on translocation of protein kinase C epsilon in cultured adult rat ventricular myocytes

AIM: To investigate the effects of mitochondrial ATP sensitive potassium (MitoKATP) channel opening on translocation of protein kinase C epsilon (PKCε) and the relationship between the translocation of PKCε and the production of reactive oxygen species. METHODS: The expression of PKCε in cultured adult rat ventricular myocytes was investigated with immunofluorescence and Western blotting techniques. RESULTS: （1） Diazoxide, a selective MitoKATP channel opener, caused a significant translocation to myofibrillar-like structures in cultured adult rat ventricular myocytes. (2) The N-2-mercaptopropionylglycine (MPG), a free radical scavenger, partly inhibited the translocation of PKCε caused by diazoxide. (3) Chelerythrine, a selective protein kinase C (PKC) inhibitor, completely blocked the translocation of PKCε caused by diazoxide. CONCLUSION: Opening of MitoKATP channel might activate PKCε and make it translocation to myofibrillar-like structures. PKCε activation occurs downstream of MitoKATP channel, and might be caused by production of reactive oxygen species after opening of MitoKATP channel.     

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.     

Expression of telomerase inhibitor Pinx1 in leukemia cells and its correlation with telomerase activity

AIM:To study the expression of telomerase inhibitor Pinx1 in acute leukemia cells and during the differentiation of acute promyelocytic leukemia cells,and to realize its effect on telomerase activity.METHODS:Realtime quantitative PCR with fluorescence probe hybridization was used to measure the expression of Pinx1 and hTERT mRNA in acute leukemia cells and during differentiation of NB4 cells induced by ATRA.The correlations between Pinx1 and hTERT expression were also analyzed.RESULTS:Pinx1 mRNA expression in acute leukemia samples (0.00312,5.42×10-4-0.024) was significantly higher than that in normal bone marrow mononuclear cells (7.89×10-4,0-0.00863,P<0.01).The expression of Pinx1 mRNA had significant positive correlation with hTERT mRNA expression (r=0.296,P<0.05).Pinx1 mRNA expression decreased during differentiation,its expression was positive correlated with hTERT mRNA expression (r=0.900,P<0.05).CONCLUSIONS:As an inhibitor of telomerase,however,Pinx1 also had the same direction of regulation with telomerase activity in acute leukemia cells,suggesting its expression variation may be a subsequent reaction induced by that of hTERT to stabilize telomerase activity.The exact mechanisms remained to be verified.     

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.     

四种杀菌剂对金银花白粉病的防治效果及对金银花品质的影响

采用随机区组试验比较四种杀菌剂对金银花白粉病的防治效果，用高效液相色谱法测定各处理样品的绿原酸含量，以筛选出符合金银花规范化种植需要的药剂。结果表明，施药后14天以15％三唑酮600倍液防治效果最好，达77．8％，其次是15％三唑酮1200倍、50％多菌灵500倍液处理，防治效果分别为75．7％和70．8％，极显著地高于3％中生菌素500和1000倍及2％农抗1201000倍液处理的防效。2％农抗120500倍液的防治效果达60．3％，与前三种处理差异不显著，显著好于后三种处理，但未达极显著水平。15％三唑酮1200倍防治处理的绿原酸含量最高，达1．683％，其次是50％多菌灵500倍处理，绿原酸含量达1．539％，符合《中华人民共和国药典》要求，可作为金银花规范化管理中白粉病防治优先采用的药剂。     

云南、福建、湖南烟区烟草花叶病主要病毒种类检测及黄瓜花叶病毒亚组鉴定

 采用抗原直接包被和双抗体夹心酶联免疫吸附测定法(ELISA)对采自云南、福建、湖南烟区烟草花叶病样品进行了病毒种类检测,利用三抗体夹心ELISA对黄瓜花叶病毒(Cucumber mosaic virus,CMV)的亚组类型进行了鉴定。在云南采集的520个花叶病样品中,烟草花叶病毒(Tobacco mosaic virus,TMV)、CMV和马铃薯Y病毒(Potato virus Y,PVY)总检出率分别为71.74%、55.01%和6.35%;在福建采集的150个花叶病样品中,TMV、CMV和PVY的总检出率分别为94%、24.66%和8.00%;在湖南采集的74个花叶病样品中,TMV、CMV和PVY的总检出率分别为58.11%、51.35%和2.70%。部分样品为2种以上病毒复合侵染。云南、福建和湖南采集的64个CMV阳性样品中,属亚组Ⅰ的样品为57个,占89.1%;属亚组Ⅱ的样品为10个,占15.6%;其中3个样品为亚组Ⅰ和亚组Ⅱ的复合侵染。     

3种粒线虫多重PCR检测方法

 剪股颖粒线虫[Anguina agrostis (Steinbuch,1799) Filipjev,1936]、小麦粒线虫[Anguina tritici (Steinbuch,1799) Filipjev,1936]和维氏粒线虫[Anguina wevelli (Van den Berg,1985) Siddiqi,2000]都是重要的植物病原线虫,它们在成熟种子和虫瘿中的虫态通常是幼虫,而这3种线虫的幼虫形态非常相似,难以根据其特征对它们进行快速准确的种类鉴定。本研究根据这3种线虫的rDNA-ITS区域序列,分别设计筛选了特异性引物AgrF₁/AgrR1、TriF₁/TriR1、WevF₁/WevR1,构建了这3种线虫单条幼虫多重PCR检测体系,获得3个大小差异明显的片段,表明这些引物设计合理,适合这3种线虫的快速准确检测。     

苇状羊茅与多年生黑麦草内生真菌菌丝基因组DNA提取的优化及PCR初步检测

感染内生真菌的禾草在牧草和草坪业上具有重要的生态和经济意义，家畜采食感染Neotyphodium coenophialum和N.lolii的苇状羊茅和多年生黑麦草会发生中毒。本研究收集天津口岸1998年以来进境的部分苇状羊茅和多年生黑麦草种子，对经镜检确认带有内生真菌的种子进行分离培养，对疑似菌株的菌丝用改进的Moiler等方法进行基因组DNA抽提，测定浓度及纯度，对照原方法，DNA的纯度有较大提高，浓度略有上升。Tubulin2基因的引物IS1-IS3扩增结果显示为单一的条带，结合形态学和序列比对，分离培养得到的菌株可以基本确定为N．coenophialum和N。lolii。根据Genbank中N.coenophialum和N.lolii的NC25基因序列设计出引物F1-R1，扩增得到能区分开N.coenophialum和N．lolii的单一条带（相差160bp），建立了N．coenophialum和N．lolii的PCR检测方法，结果准确可靠。     

浙江杭州地区蝶类资源及区系分析

杭州地区有蝶类113种,隶属于12科76属。其中古北东洋共有种48种,东洋种42种,古北种17种,广布种5种。区系分析结果表明,杭州地区蝴类以古北东洋共有种和东洋种为主要成份,属东洋区系范畴,并具有由东洋区向古北区过渡的特征。与江、浙其他地区的蝶类资源进行相似度分析,杭州地区与宁波地区的蝶类资源相似度最高。