Evaluation of ecosystem services of Chinese pine forests in China

Evaluation of forest ecosystem services is a hot topic,both in China and at abroad,but it has not yet obtained a consistency of evaluation indicator systems and evaluation methods.Under the framework of evaluation criteria to be implemented for forest ecosystem services,years of consecutive observation data from Long Term Ecological Research Stations affiliated to Chinese Forest Ecosystem Research Network(CFERN),forest resource inventory and public data were applied to carry out a detailed and dynamic evaluation on the physical quantity and value of ecosystem services of Chinese pine forests in China.The results showed that the above services had the total value and unit value of 1144.9640 billion(1.1449640×10 12 )RMB and 52.074 thousand RMB per hectare per year,respectively during the 9th Five-year Plan(1996―2000),and of 1190.5461 billion RMB and 52.101 thousand RMB per hectare per year,respectively,during the 10th Five-year Plan(2001―2005).For Chinese pine forests,water conservation was 40.40 hundred million cubic meters annually,soil conservation was 67 million tons and C fixation 9 million tons annually,production of healthful negative ions was 1.96×10 20 , absorption of SO2 was 5.02 hundred million kilograms and dust-catching was 759.10 hundred million kilograms. Among the 15 provinces of China with Chinese pine forests,the biggest beneficiary from ecosystem services was Liaoning Province;while Hunan Province was the smallest beneficiary between the 9th Five-year Plan.     

Ablation of Promyelocytic Leukemia Protein (PML) Re-patterns Energy Balance and Protects Mice from Obesity Induced by a Western Diet

The promyelocytic leukemia protein is a well known tumor suppressor, but its role in metabolism is largely unknown. Mice with a deletion in the gene for PML (KO mice) exhibit altered gene expression in liver, adipose tissue, and skeletal muscle, an accelerated rate of fatty acid metabolism, abnormal glucose metabolism, constitutive AMP-activating kinase (AMPK) activation, and insulin resistance in skeletal muscle. Last, an increased rate of energy expenditure protects PML KO mice from the effects of obesity induced by a Western diet. Collectively, our study uncovers a previously unappreciated role of PML in the regulation of metabolism and energy balance in mice.     

抗IFITM1单克隆抗体的制备及检测

目的：制备抗干扰素诱导的跨膜蛋白-1（interferon-induced transmembrane protein 1,IFITM1）的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法：以结肠癌患者的癌组织为材料,提取总RNA,以RT—PCR扩增得到IFITM1 cDNA序列,经EcoR 和HindⅢ双酶切后,克隆入pGEX-4T-3进行原核表达并纯化得IFITM1-GST;以该融合蛋白免疫BALB／c小鼠,淋巴细胞杂交瘤法制备单克隆抗体;采用ELISA、Western-blot及免疫组织化学法以制备的抗体检测结肠癌患者结肠癌组织中的IFITM1。结果：成功构建了1FITM1核表达载体,获得了IFITM1-GST重组蛋白;制备得到了1株抗IFITM1单克隆抗体,腹水ELISA效价为1：30000,抗体亚类为IgG1,可用于ELISA、Western-blot及免疫组织化学法检测结肠癌患者结肠癌组织中的IFITM1。结论：获得了1株可用于ELISA、Westem-blot及免疫组织化学法的抗IFITM1单克隆抗体2F—1,为进一步研究IFITM1在结肠肿瘤发生过程中的作用提供了实验基础。     

H1亚型猪流感病毒血凝素基因在昆虫细胞中的表达及其间接ELISA方法的初步建立

根据GenBank发表的H1亚型猪流感HA基因序列设计引物,扩增出HA基因片段.将其克隆到pFastBacGP67B杆状病毒载体上,筛选阳性重组转座载体pFastBacGP67B-H1,转化含有杆状病毒穿梭载体(bacmid)的DH10Bac感受态细胞,构建杆状病毒表达载体获得重组转座子(rBacmid-H1),在脂质体介导下转染sf9昆虫细胞,获得重组杆状病毒(rBV-H1),再感染细胞,收获目的蛋白.通过血凝试验、免疫印迹法、免疫组化分析表明该蛋白得到表达,且具有良好的生物学活性.利用表达的蛋白作为猪流感间接ELISA的抗原,初步建立H1亚型猪流感的间接ELISA检测方法,并对内蒙古、辽宁和黑龙江等地送检的93份猪血清进行了检测,阳性率为31.18%,为研制开发快速、准确、简便的H1亚型猪流感鉴别诊断试剂盒奠定基础.     

苹果树腐烂病菌细胞色素P450基因Vmcyp5的功能

【目的】研究表明,细胞色素P450(CYP)在死体营养型真菌的毒素合成代谢中发挥重要作用,预测可能与病原菌致病相关。论文对苹果树腐烂病菌(Valsa mali)毒素合成基因簇中的1个上调表达的CYP基因Vmcyp5进行生物学功能研究,明确CYP基因对病原菌致病力影响,为细胞色素P450基因家族对苹果树腐烂病菌致病机理的进一步研究提供依据。【方法】通过Double-joint PCR和PEG介导的原生质体转化技术获得具有G418抗性的突变体,并对突变体进行PCR检测及Southern blotting验证得到单拷贝敲除突变体。将目的基因片段重新导入敲除突变体,筛选获得互补突变体。最终对野生型菌株及敲除突变体、互补突变体进行菌落、产孢及致病力观察,利用SPSS软件对数据进行差异显著性分析,并利用q RT-PCR技术分析突变体黑色素基因簇的表达水平。【结果】通过基因敲除技术获得1个Vmcyp5基因的敲除突变体。与野生型菌株相比,Vmcyp5基因的敲除突变体菌落呈白色,产孢量减少51.3%。q RT-PCR分析发现敲除突变体黑色素基因簇基因表达量降低。重要的是,敲除突变体致病力较野生型菌株降低24.5%。互补突变体菌落颜色、产孢及致病力近似恢复至野生型菌株水平。【结论】Vmcyp5基因与病原菌黑色素合成、子实体的产生和致病力相关。     

猴D型反转录病毒巢式PCR检测方法的建立

目的建立SRV-1巢式PCR检测方法并进行初步应用。方法针对SRV-1env基因的保守区序列,设计特异性引物,以感染SRV-1 Raji细胞提取出的含有前病毒DNA的基因组DNA为模板,进行巢式PCR反应。扩增产物测序后与GenBank报道的序列进行同源比对。将DNA样本进行10倍梯度稀释,以检测巢式PCR反应的灵敏度。使用该方法对正常Raji细胞以及感染SIV、STLV的外周血淋巴细胞DNA样本进行扩增,检测该方法的特异性。用建立的巢式PCR方法检测40份储存猴血标本。结果使用巢式PCR扩增出的特异片段经测序分析,结果证实与GenBank报道的序列一致。所建立的巢式PCR检测法检测限度可达1.5×10-3ng/μL,而且方法特异。用此方法检测40份猴血标本,未检测到阳性标本。结论初步建立SRV-1的巢式PCR检测方法,该方法灵敏、特异,为SRV-1的检测提供了一个快速、有效的手段。     

重组人白细胞介素6的制备

我们采用本院基础医学研究所组建的ＩＬ-６工程菌Ｅ．ｃｏｉｌＤＨ５ａ（ｐＢＶ-ｈｌＬ－６）,在选定的培养基及ｐＨ值下,采用３０升发酵罐进一步观察了工程菌的生长和ｒＩＬ－６的表达,确定了发酵工艺条件。在此条件下连续进行了三批次的培养试验。结果表明,工程首的生长密度达到２．５１±０．０２ｇ［干重］／Ｌ［发酵液］,ｒＩＬ-６产率为１８２．４±２．０ｍｇ／ｇ［干重］。ｒＩＬ-６以包涵体形式表达于大肠杆菌细胞中,破菌后选用非离子型去垢剂或尿素等变性剂提取包涵体中的杂蛋白,可使ｒＩＬ-６的纯度达到７０．１±１．３％,收率为７１．９±１．９％。洗涤后的包涵体,经过凝胶柱纯化和复性,ｒＩＬ-６纯度达到９５％以上,柱纯化的收率为７２．３±０．９％;采用依赖ＩＬ-６,小鼠杂交瘤细胞系７ＴＤ１及ＭＴＴ比色法测定生物活性,ｒＩＬ-６比活性达２×１０８Ｕ／ｍｇ。     

Simultaneous quantitation of dexamethasone palmitate and dexamethasone in human plasma by liquid chromatography/tandem mass spectrometry

A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for simultaneous quantitation of dexamethasone palmitate and dexamethasone in human plasma was developed. After sample preparation by protein precipitation and liquid-liquid extraction, the analytes and internal standard (IS) were separated on a Venusil XBP-C8 column using gradient elution. Multiple reaction monitoring of dexamethasone palmitate, dexamethasone and IS used the precursor to product ion transitions at m/z 631.8-->373.1, m/z 393.2-->147.1 and m/z 264.2-->58.1, respectively. The method was linear over the ranges 1.5-1000ng/mL for dexamethasone palmitate and 2.5-250ng/mL for dexamethasone with intra- and inter-day precisions of <10% and accuracies of 100+/-7%. The assay was applied to a clinical pharmacokinetic study involving the injection of dexamethasone palmitate to healthy volunteers.     

地塞米松对HepG2细胞分泌载脂蛋白的影响

以培养的人肝癌细胞系HepG2为研究对象,采用“冻干浓缩培养液载脂蛋白测定法”,考察了地塞米松对HepG2细胞载脂蛋白(apolipoprotein,apo)AⅠ、AⅡ、CⅢ、B100及E分泌的影响.结果表明：地塞米松对HepG2细胞apoAⅠ和apoE的分泌有促进作用,对apoAⅡ、apoB100和apoCⅢ的分泌有抑制作用,且这种作用随地塞米松浓度的增加而增强.当培养液中地塞米松的浓度为5.5×10^－5mol/L时,apoAⅠ和apoE的分泌分别增加36.6%和49.4%（P＜0.01）,apoAⅡ、apoB100和apoCⅢ的分泌分别减少38.9%、31.9%和29.8%（P＜0.01）.