The ex vivo expansion characteristic of endothelial progenitor cells

AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34⁺ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34⁺ and CD34^- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34⁺ and CD34^-,total MNC led to a significant increase in the expansion of CD34⁺ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34⁺, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34⁺ and AC133⁺cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34⁺ and CD34^- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.     

Influence of Sini decoction on the proliferation and apoptosis of artery smooth muscle cells after balloon injury

AIM: To investigate the influence of Sini decoction (SND) on the proliferation and apoptosis of rabbit abdominal aorta smooth muscle cells after ballon injury and discuss the effect of vascular smooth muscle cell's (VSMCs) proliferation and apoptosis in post-percutaneous coronary intervention (PCI) restenosis (RS) and the feasibility of SND preventing post-PCI RS. METHODS: The animal model of rabbit abdominal aorta ballon injury was set up and the therapertic group was treated with SND. The shape of proliferative and apoptotic cell were investigated by electron microscope. Immunohistochemistry staining was performed using α-actin,PCNA and Cyclin E monoclonal antibodies. In situ Cell Death Detection Kit was used to identify apoptotic cells. Abdomial aorta angiography was operated in the 84th day subgroup and the stenosis degree was evalued by quantitative angiographic analysis. RESULTS: As compared with the control group, the therapeutic group displayed a lower proliferative percentage and a higher apoptosic percentage (P<0.05). Moreover, the apoptosic peek time was on the 14th day after operation,which was longer than the control group. CONCLUSION: SND effectly inhibited the proliferation of VSMCs and iuduced apoptosis in VSMCs.     

Effects of Sini decoction on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion

AIM: To investigate the effect of Sini decoction (SND)on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion. METHODS: Thirty-two Sprague-Dawley rats of both sexes were randomly divided into 4 groups: (1) Control group in which sham operation was performed; (2) Model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND1 group in which SD (0.6 g/200 g rat) was given via stomach tube 3 d before intestinal I/R; (4) SND2 group in which SD (1.2 g/200 g rat)was given via stomach tube 3 d before intestinal I/R. A strip of small intestine was taken from distal end of ileum for electron microscopic examination. The two-dimensional structural parameters and three-dimensional structural parameters of mitochondria were calculated. RESULTS: (1)Morphological changes of small intestine: In control group, epithelial cells were orderly arranged, with normal mitochondria and intestinal villi. In model group, the gaps between epithelial cells widened. There were a lot of apoptotic cells. Microvilli were short and swelled. Mitochondria were swelled obviously with broken ridges. Endoplasmatic reticulum was severely dilated. In SND1 and SND2 groups, microvilli and epithelial cells were orderly arranged relatively, mitochondria was slightly swelled. (2) Structural parameters of mitochondria: In model group, there were the least mitochondria and the swelling of mitochondria was severe. In SND1 and SND2 groups, the mitochondria was more than that of model group and the swelling were slight. CONCLUSION: Sini decoction can protect small intestine from ischemia-reperfusion injury without dose-dependent effect.     

Changes of MAPKs expression in rat hippocampal neurons after sleep deprivation

AIM: To explore the change and the possible role of MAPKs in rat hippocampus neuron after sleep deprivation. METHODS: The morphology of hippocampus neuron after sleep derivation was observed by TUNEL and HE staining, the activity of ERK was assayed by β-liquid scintillation counting and the expression of JNK was detected by Western blot. RESULTS: In paradoxical sleep deprivation (PSD) group, the number of apoptotic cells in hippocampus was increased. The scores of ERK activity were 1 764.00±941.56. Compared with control groups, the ERK activity was obviously decreased (P<0.05). The JNK expression was 87.5%, which was higher than that in control group. CONCLUSION: These results provide some important evidences that the sleep deprivation could cause changes in MAPKs activity, which may be related to the mechanism of hippocampus neuron apoptosis.     

The role of resistin in obesity and diabetes

Increased evidences indicate that resistin is a new hormone secreted from adipose tissue, it is reported to be an important signal molecule linking between obesity, insulin resistance and type 2 diabetes. Many factors can affect the gene expression of resistin. However, the detailed function of resistin still remains mysterious and much work needs to be undertaken.     

规模化猪场大肠杆菌的耐药性监测及血清流行病学调查

为了调查猪致病性大肠杆菌的耐药性及流行血清型 ,从湖北、河南、江西、安徽、浙江等省的 33个猪场采集 32 1份仔猪黄、白痢病料进行细菌的分离和生化鉴定 ,结果分离到大肠杆菌 30 2株 ,其中致病性大肠杆菌 2 76株。对 112株致病性大肠杆菌进行了 15种抗生素敏感性试验 ,发现分离菌株对 15种药物均有不同程度的耐药性 :青霉素 G对其完全没有抑制作用 ;先锋霉素 抑制作用最强 (97.3% ) ,其次为呋喃妥因 (78.6 % )、痢特灵 (71.4 % )、环丙沙星(6 8.8% )、诺氟沙星 (6 5 .1% )。 112株菌中 ,有 4 7种耐药谱型 ,多数为 6耐以上的菌株 (80株 ) ,占供试菌株的 71.4 %。应用微量平板凝集试验 ,对分离的 2 76株致病菌进行了血清型鉴定 ,鉴定共有 72种血清型 ,其中优势血清型 10种 ,占能定型分离致病菌株的 5 3.7%。     

鸡痘病毒通用高效表达载体的构建及其初步应用

利用分子克隆技术对本室构建的高效鸡痘病毒表达载体 p1 1 S进行改造 ,在其人工合成禽痘病毒 ( FPV)强启动子 Ps的下游引入含 7个单一酶切位点的多克隆位点 ( MCS) ,构建了便于外源基因插入的通用性更强的高效单表达载体 p N1 1 S。然后将 FPV早晚期启动子 PE/ L 及其启动的鹅源新城疫病毒 NDV ZJ1株的 F基因一并插入 p N1 1 S中 ,使PE/ L 与 Ps反向串联 ,从而构建出 1个含 NDV ZJ1株 F基因的重组 FPV高效双表达载体 p N1 1 SEF,其 MCS可以用于插入其他外源基因。在此基础上 ,将 NDV ZJ1株的 HN基因插入 p N1 1 SEF的 Ps启动子下游的 MCS中 ,构建了共表达 NDV ZJ1株 F和 HN基因的重组 FPV双表达载体 p N1 1 SEFHN。将 p H1 1 SEFHN质粒 DNA与 FPV2 82 E4株共转染 CEF,得到共表达 NDV ZJ1株和 HN基因的重组 FPV,间接免疫荧光实验初步证明外源基因得到了较好的表达。表明所构建的通用高效 FPV表达载体有利于高效基因工程活载体疫苗的研制 ,具有广阔的应用前景     

生长肥育猪骨骼肌注射表达pGRF基因质粒的效应研究

将猪的GRF基因表达质粒注射于猪的骨骼肌后,研究其促生长效应与机理。选用始重6.3kg的44头去势长白×太湖仔猪,分6组,采用2×3因子安排的完全随机区组设计,按6～10kg、10～20kg、20～50kg、50～100kg4个阶段饲养。4个饲养阶段的饲粮低蛋白水平分别是20.70%,17.90%,15.03%,13.00%;高蛋白水平分别是23.70%,20.90%,18.02%,16.00%。pGRF基因质粒注射剂量设0mg、0.5mg、1.0mg3个水平,于试验开始与试验猪体重达60kg时共注射基因质粒2次。测定各阶段日增重,饲料消耗量,耗料增重比以及30、70、100kg时血液中GH、GRF、IGF-I的浓度。100kg时屠宰进行胴体品质测定。结果表明:饲粮蛋白水平对各阶段及全期试验猪日增重均有显著影响(P<0.05或P<0.01),对50～100kg阶段与全期日采食量和耗料增重比有显著影响(P<0.05或P<0.01)。注射pGRF基因质粒对各阶段及全期日增重均有显著影响(P<0.05),对6～10kg、10～20kg、50～100kg3阶段及全期日采食量有显著影响(P<0.05),对6～10kg阶段、50～100kg阶段及全期耗料增重比有显著影响(P<0.05)。注射pGRF基因质粒对30kg及70kg体重时猪血液中GRF、GH、IGF-I浓度均有显著影响(P<0.05)。提高饲粮蛋白水平与注射pGRF基因质粒均可明显降低超声波测膘厚及屠宰测膘厚、增大眼肌面积(P<0.05)。     

Comparative studies of acetylcholinesterases purified from two field populations of the oriental migratory locust (Locusta migratoria manilensis): implications of insecticide resistance

Acetylcholinesterase (AChE) was purified by affinity chromatography from two populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen), collected from Huanghua and Pingshan Counties, Hebei Province of China. The purification factors and yields were 1661-fold and 19.3%, respectively, for the Huanghua population, and 3897-fold and 39.6% for the Pingshan population. Both the purification factor and yield were significantly lower in the Huanghua population than in the Pingshan population. AChE activity was almost completely inhibited by 10⁻⁶ M eserine and BW284C51, but ?5.8% of AChE activity was inhibited by ethopropazine at the same concentration, suggesting that purified AChE from either population was a typical insect AChE. However, AChE purified from the Huanghua population was 62-, 2.0-, and 1.6-fold less sensitive to inhibition by the three organophosphate compounds, chlorpyrifos oxon, demeton-S-methyl, and paraoxon, respectively, than that from the Pingshan population. Significantly lower purification factor and low yield associated with reduced sensitivity of AChE to inhibition by the organophosphates indicated that AChE purified from the Huanghua population was biochemically and pharmacologically different from that of the Pingshan population. Reduced sensitivity of AChE appeared to contribute to organophosphate resistance in the locust from Huanghua County, where insecticides have commonly been used to manage outbreaks of the locust.     

华北农牧交错带退化草地土壤种子库动态变化

对华北农牧交错带不同退化演替阶段土壤种子库的数量和物种组成及与地上植被的关系进行了初步的研究,结果表明:农牧交错带草地在刈割和放牧等干扰下,发生逆行演替,土壤种子库中植物种类及种子数量呈递减的趋势,轻度退化类型土壤种子库物种组成最高,有23种植物,种子数量最多,有4 303粒,禾本科所占比例较大,为32.3%;而极度退化草地物种组成最低,有14种植物,种子数量仅有1 156粒,禾本科只占8.4%.草地逆行演替过程中植被与土壤种子库在物种组成上存在一定差别,中度退化阶段差别最大,有18种植物未在种子库中出现,极度退化阶段差别最小,有2种未在种子库中出现.