我国农村木材市场启动升温

2005年我国政府对经济发展形势实行宏观调控,在财政和金融方面采取了"双稳"的政策.     

三分域模型在研究单胃动物内源氨基酸排泄中的应用

本文在分析三分域链状系统模型及单胃动物内源氨基酸排泄特点的基础上 ,讨论了如何将三分域链状模型用于单胃动物内源氨基酸排泄模式的确定。研究分三步进行 :1.测定动物血液体积 ;2.静脉连续灌注同位素标记的氨基酸并使之于体内达到平衡 ;3.检测消化道中同位素标记氨基酸的排泄率。文章指出 ,若能对所有必需氨基酸的内源排泄率均进行测定 ,则可得出内源氨基酸排泄模式 ,因而在实际测定饲料或日粮真可消化氨基酸含量时 ,只需测定某种氨基酸的内源排泄量即可知其它氨基酸的内源排泄状况     

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck, one of the most popular edible mushrooms, has attracted great attention due to its delicious taste and healthy properties. However, both its biological effects and the possible mechanism of action have not yet been known. We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck (MPE) against HepG2 human hepatocellular carcinoma cells. Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90% (P<0.01). The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax, Bcl-2 and cleaved caspase-3. On the other hand, MPE might trigger cell cycle arrest at G₀/G₁/S phases by managing p21, Cyclin D1, cyclin-dependent kinases-4 (CDK4) and proliferating cell nuclear antigen (PCNA). Additionally, MPE downregulated TRAF-2 and p-p53, while upregulated p-ASK1 and p-p38. Therefore, it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.     

Comparison of phenolic profiles and antioxidant activities in skins and pulps of eleven grape cultivars(Vitis vinifera L.)

Eleven grape cultivars were analysed to explore the variety differences of fresh grape phenolic profiles. The results showed that free phenolics were predominant in grape skins and pulps, and showed the higher antioxidant activities than bound. In 11 cultivars, Muscat Kyoho extracts had the highest total phenolic content in skins(10.525 mg GAE g~(–1) FW) and pulps(1.134 mg GAE g~(–1) FW), and exhibited the highest DPPH radical scavening capacity(EC_(50)=11.7 μg mL~(–1)) and oxygen radical absorbance capacity(ORAC) value(190.57 μmol TE g~(–1) FW) of free phenolic in skin. In addition, the most abundant phenolics in grape skins were found to be flavonoids such as kaempferol in Kyoho skin(541.2 μg g~(–1) FW), rutin, catechin and epicatechin in Muscat Kyoho skin(262.3, 86.3 and 70.0 μg g~(–1) FW, respectively). Furthermore, the principal component analysis showed a strong difference of phenolic profiles with the cultivars, existing forms and distributions. Pearson correlation coefficient analysis showed a significant linear correlation between total phenolic content and antioxidant activity(P0.05). Therefore, both skins and pulps were rich sources of bioactive phenolic compounds, and Muscat Kyoho was the ideal source among all samples.     

SMART技术构建的辣椒疫霉菌与辣椒互作AD-cDNA文库

为了分离和克隆辣椒中疫霉菌诱导基因,以接种辣椒疫霉菌的叶片为材料,利用SMART技术构建辣椒疫霉菌与辣椒互作的双杂交c DNA文库。结果表明:该文库容量为3.6×106cfu,重组率88%左右,插入片段集中在300~2000bp之间,平均长度约为800bp,表明获得的文库质量较高,该文库将为分子育种提供重要的基因资源。     

菌丝霉素分泌型丁酸梭菌防治罗非鱼链球菌病的应用研究

为解决罗非鱼链球菌感染难题,本研究在抗菌肽菌丝霉素分泌型丁酸梭菌制剂的基础上,对其进行了抑菌活性、稳定性和罗非鱼养殖应用研究。结果发现:表达产物菌丝霉素对常见致病菌如大肠杆菌、沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌和猪链球菌均有较好的抑菌活性。选用米糠作为菌丝霉素分泌型丁酸梭菌的保护剂载体,此制剂可在常温保存一年。通过上述的应用基础试验,初步建立一套菌丝霉素分泌型丁酸梭菌制剂养殖应用方案,此方案可显著降低罗非鱼链球菌病发病率58.6%以上。本研究为丁酸梭菌制剂在防治罗非鱼链球菌病应用方面提供了参考,此制剂有望替代抗生素,促进罗非鱼养殖业的持续健康发展。     

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒(PRV)在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞(3D4/21)p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

6个果桑品种主要性状调查与染色体倍性测定

作者对黑龙江省6个果桑品种资源进行性状调查和染色体观察。结果表明,‘龙桑1号’具有染色体数为28和42的细胞,是四倍和六倍的混合体;‘选2’为自然六倍体;‘龙椹1号’‘选3’‘选4’‘矮化1号’均为四倍体。该研究对高寒地区桑树资源的发掘与创新利用具有重要意义。     

CD147 monoclonal antibody-mediated nanoparticles for gene therapy to target lung cancer cells

AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells. The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed. METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector. The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope. The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group. Non-transfected cells were used as control group. The cell transfection was carried out with 250 ng plasmids/well in 6-well plate. The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope. The mRNA expression of PKCε was detected by RT-qPCR. The protein expression of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot. The cell proliferation ability was detected with colony formation assay. The cell invasion ability was detected by Transwell method. RESULTS: The expression of CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining. The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76±0.18)%, (98.51±0.32)%, (99.17±0.16)% and (99.68±0.11)%, respectively. The difference between CP group and control group was statistically significant (P<0.05). No significant difference among CN group, LP group and control group was observed. The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference. The colony number in CP group was significantly smaller than that in control group (P<0.05). The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group. The number of the invading cells in CP group was significantly less than that in control group (P<0.05). The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group. CONCLUSION: Nanogene vector targeting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibitory effects on the proliferation and invasion of the lung cancer cells.