毛木耳多糖的药理研究进展

毛木耳为我国传统大型食药用真菌,目前认为毛木耳多糖是毛木耳的主要活性成分之一。现代药理学研究表明,毛木耳多糖对心血管系统有一定的影响作用,同时具有增强机体免疫、抗氧化、抑制肿瘤等广泛的药理作用。就近年来有关毛木耳多糖药理作用的研究进行综述。     

Effect of BDNF expression in cerebral cortex and hippocampus on ability of learning and memory in APP/PS1 transgenic mice

AIM: To investigate the expression changes of brain-derived neurotrophic factor (BDNF) in the cerebral cortex and hippocampus and their effects on the ability of learning and memory in the wild-type (WT) mice and APP/PS1 transgenic mice. METHODS: WT mice and APP/PS1 transgenic mice were selected as study subjects. Aβ plaques, apoptosis rate and BDNF expression in the cerebral cortex and hippocampus of WT mice and APP/PS1 transgenic mice were detected by the methods of Congo red staining, TUNEL, immunofluorescence and Western blot. The abilities of learning and memory were determined by Morris water maze test. RESULTS: The Aβ plaques appeared in the cerebral cortex and hippocampus of APP/PS1 transgenic mice, and the number of Aβ plaques in 12-month-old mice was larger than that in 6-month-old mice (P<0.05). The number of apoptotic neurons in the cerebral cortex and hippocampus of 12-month-old APP/PS1 transgenic mice was larger than that of WT mice (P<0.01). The expression level of BDNF in the cerebral cortex and hippocampus of WT mice was higher than that of APP/PS1 transgenic mice (P<0.01). The Morris water maze test showed that the escape latency in APP/PS1 transgenic mice was longer than that in WT mice, and the times across the platform quadrant in 60 s was less than that in WT mice (P<0.01). The swim-tracking path of APP/PS1 transgenic mice was disordered and irregular. CONCLUSION: The expression of BDNF in the cerebral cortex and hippocampus of APP/PS1 transgenic mice was lower than that of WT mice, accompanied by increased neuronal apoptosis and decreased spatial learning and memory ability. The decrease in learning and memory ability may be related to decreased BDNF expression in the cerebral cortex and hippocampus of APP/PS1 transgenic mice, leading to increased neuronal apoptosis, which may be one of the pathological mechanisms of Alzheimer disease.     

纳米SiO2 -APP 对木塑复合材料界面特性及力学性能的影响

用聚磷酸铵(APP)与阻燃协效剂纳米二氧化硅(SiO2 )制备了阻燃型木塑复合材料,并利用FTIR、SEM 和力学 性能测试仪,探讨纳米SiO2 的添加量(2%、4%、6%)和APP 的添加量(8%、10%、12%)对木塑复合材料的界面性 能和力学性能的影响。结果表明:1)当纳米SiO2 添加量为2% ~6%、APP 添加量为8% ~10%时,两者可以均匀地 分布在木塑复合材料的孔隙中,并且纳米SiO2 可以与木质纤维形成Si—O—C 结合,改善复合材料的界面性能;但 是,APP 添加量增加至12%时,纳米SiO2 和APP 之间会发生团聚,降低了复合材料的性能。2)当纳米SiO2 添加量 为2% ~6%、APP 添加量为8% ~10%时,木塑复合材料的拉伸强度和弯曲强度均比未添加纳米SiO2 、APP 的有所 增加,拉伸断裂伸长率基本保持不变,冲击强度降低。通过双因素方差分析可知,纳米SiO2 、APP 的添加量以及两 者之间的交互作用对拉伸性能、弯曲性能无显著影响,但APP 的添加量以及两者之间的交互作用对冲击强度有显 著影响。      

APP蛋白家族胞内段释放与神经干细胞向神经元细胞定向分化的关系研究

[目的]研究APP家族胞内段对神经细胞定向分化的影响。[方法]通过分离体外培养神经干细胞,转染导入APP蛋白家族胞内段以过表达后,研究其对神经干细胞向神经元定向分化的影响。[结果]APP家族蛋白胞内段的过表达可以抑制神经干细胞向神经元细胞的分化,其中APP和APLP2胞内段的影响尤为显著。突变试验表明,通过Capase水解释放末端31氨基酸片断是这个过程必需的。APP家族胞内段是通过释放APP-ICD31来抑制神经干细胞向神经元细胞的定向分化。[结论]该研究进一步证实了APP蛋白和老年痴呆病发生的联系,也为老年痴呆症的治疗提供新的靶点。     

广东省猪传染性胸膜肺炎放线杆菌分离鉴定及耐药表型和耐药基因检测与分析

【目的】了解广东省部分规模化猪场引起猪传染性胸膜肺炎(PCP)的病原菌猪传染性胸膜肺炎放线杆菌(APP)的耐药表型和耐药基因携带情况,并分析比较其相关性,为有效防控该病提供理论依据。【方法】将2019-2021年从广东省不同地区规模化猪场采集的疑似猪传染性胸膜肺炎病死猪病料通过病原分离培养、革兰氏染色、生化试验、PCR扩增等方法进行病原菌分离鉴定和测序分析,随后对分离株进行药敏试验并通过PCR检测其耐药基因,确定其耐药表型和耐药基因的携带率,分析比较两者的一致性。【结果】分离菌需在含有血清和NAD的培养板中生长;革兰氏染色结果显示分离菌为红色,可判定为革兰氏阴性细菌,通过生化试验、PCR结果及测序分析确定分离到20株APP,且没有表现出明显的地域特性。所有菌株均呈现多重耐药且约50%菌株呈8重及以上耐药;其中对四环素类、磺胺类、氯霉素类和大环内酯类药物耐药率较高,分别为77.5%、65.0%、55.0%和48.8%;进一步分析发现,分离菌主要对四环素、磺胺异噁唑、氟苯尼考和林可霉素等抗菌药物耐药,而对头孢唑啉(先锋Ⅴ)和阿奇霉素敏感。23种主要耐药基因中共检测到bla_CMY、aph(2″)-Ⅰb、sul1、sul2、sul3、tetA、tetB、tetM、tetO、tetR和floR这11种耐药基因,携带率分别为85%、85%、50%、60%、75%、30%、100%、85%、100%、70%和80%;未检测到喹诺酮类、大环内酯类和林可胺类相关耐药基因。【结论】从广东省猪群中分离到的APP表现出广泛耐药性,且为多重耐药,耐药基因与耐药表型基本一致,表明耐药基因的携带是细菌对抗菌药物产生耐药的主要原因之一,但也可能存在其他未检测的耐药基因,或存在新的耐药机制。     

多重耐药胸膜肺炎放线杆菌GD2107株全基因组测序及生物信息学分析

【目的】通过对多重耐药胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)GD2107株进行全基因组测序及生物信息学分析,丰富APP基因组数据库信息;构建基于ApxⅣ基因的系统进化树,分析该菌株的进化关系,为探索APP致病机制和临床防控猪传染性胸膜肺炎(porcine contagious pleuropneumia,PCP)提供参考。【方法】通过药敏试验测定分离菌株的耐药谱;采用全基因组测序技术(whole genome sequencing,WGS)对细菌DNA进行全基因组测序,分别利用Illumina NovaSeq、PacBio SequeI测序平台对全基因组测序结果进行基因功能注释及生物信息学分析(包括基因组基本信息、功能元件分析及亚系统分析等);基于ApxⅣ基因构建系统进化树。【结果】药敏试验结果显示,GD2107菌株对青霉素、头孢拉定(先锋Ⅵ)、卡那霉素等14种抗菌药均耐药。对GD2107株全基因组测序得到1条大小为2 271 987 bp的环状染色体(GC含量为41.21%)和2个大小分别为5 027和3 497 bp的环状质粒,共预测到2 290个编码基因,包含19个rRNA (7个5S rRNA、6个16S rRNA、6个23S rRNA)、21个tRNA基因、20个ncRNA;23个基因岛、4个原噬菌体和2组CRISPR相关序列;分别有2 112、1 549和1 866个基因在COG、KEGG和GO数据库中得到注释,且相关蛋白集中分布于APP的代谢过程;另外在毒力因子(VFDB)和耐药因子(CARD)数据库中还注释到48个毒力基因和22个耐药基因(仅floR基因位于质粒上)。绘制该菌株的全基因组圈图,将基因组信息提交至NCBI,获得染色体GenBank登录号为CP097377,质粒登录号分别为CP097378和CP097379。系统进化树分析发现,该菌株与来自中国的APP菌株(CP063424.1)进化关系最近。【结论】本研究完成了对多重耐药菌株GD2107的全基因组测序与生物信息学分析,全面认识了该菌株基因组的结构和功能并探究了耐药和致病机制中的相关基因,进化关系显示该菌株具有一定的地域流行性,为预防PCP的流行和探索APP的致病机制提供了参考。     

Effects of Chinese herbal compound Huannaoyicongfang on inflammatory cytokines and oxidative stress in brain of APP transgenic mice

AIM: To study the effects of Chinese herbal compound Huannaoyicongfang (HNYCF) on inflammatory reaction and oxidative stress in the brain of Alzheimers disease (AD) animal model, and to explore its role in treating AD. METHODS: APP695V717I transgenic mice (3 months old),as an AD model in this study, were randomly divided into model group, donepezil group, HNYCF high-dose group and HNYCF low-dose group. C57BL/6J mice, which were of the same age and genetic background as the transgenic mice, were used as controls. The animals were administered intragastrically with the drug or water from 3 months old to 9 months old. Morris water maze test was performed to measure the spatial learning and memory ability. Step-down test was performed to observe the learning and memory ability of single passive avoidance response. The expression of nuclear factor kappa B (NF-κB) and peroxisome proliferator-activated receptor γ (PPARγ) in hippocampus CA1 region was detected by immunohistochemistry with image analysis. The levels of interleukin-6 (IL-6) and high sensitivity C-reactive protein (hs-CRP) in the brain cortex and hippocampus homogenate were measured by radioimmunoassay. The activity of superoxide dismutase (SOD) in serum was detected by colorimetry. The content of malondialdehyde (MDA) in serum was detected by thibabituric acid method. RESULTS: Morris water maze test showed that the times of crossing platform, and the swimming time and distance in the fourth quadrant in HNYCF groups were much more than those in model group. The step-down test manifested that the escape latency in HNYCF high-dose group was significantly longer than that in model group. Compared with model group, the expression of NF-κB obviously decreased, the expression of PPARγ significantly increased and the content of IL-6 was lower in HNYCF groups. The activity of serum SOD in HNYCF groups was significantly higher than that in model group. CONCLUSION: HNYCF evidently ameliorates the learning and memory ability in APP transgenic mice, which may be related to the anti-inflammatory and anti-oxidation effects of HNYCF.     

Expression of α-synuclein with aging in APP transgenic model of Alzheimers disease

AIM: To investigate the expression of α-synuclein in the brain of AD-like animal model at different age and to explore the pathology mechanism of α-synuclein in neural degeneration.METHODS: APP V717I transgenic (Tg) mouse model of Alzheimers disease was observed at age of 4,10 and 16 months.The Tg mice were randomly divided into 3 model groups (4,10 and 16 months of age).Control adopted the same age and background C57BL/6J mice.The mRNA expression of α-synuclein was measured by genechips and RT-PCR method.The protein of α-synuclein was detected by immuno-histochemistry and Western blotting.RESULTS: The expression of α-synuclein mRNA increased significantly in hippocampus and cortex in Tg mice at age of 4 months,10 months and 16 months compared with age matched controls.The production of α-synuclein protein also increased significantly in hippocampus and cortex in Tg mice at 3 groups.With aging,α-synuclein expression was increased and aggregated in Tg mice.CONCLUSION: Overexpression and aggregation of α-synuclein in different age of APP transgenic mice may play a key role in learning-memory deficit and the pathology of Alzheimers disease,aging,and neural degeneration.     

猪传染性胸膜肺炎PCR诊断方法的建立

根据已发表的猪胸膜肺炎放线杆菌APXIV毒素的基因序列，自行设计和合成了二对可扩增448bp和365bp目的片段的引物，成功的建立了检测APP的套式PCR方法。通过对猪肺疫巴氏杆菌、猪链球菌、大肠杆菌、猪嗜血杆菌、猪肺炎支原体和猪丹毒杆菌的DNA进行了PCR检测，结果均为阴性；对猪胸膜肺炎放线杆菌的1、2、5、6、7、9国际标准血清型均扩增出448bp和365bp的特异性条带；检测的敏感度一步PCR可达到5OO个细菌，最低检出DNA浓度可达到0．585ng／mL；套式PCR可达到50个细菌，最低检出DNA浓度可达到58．5pg／mL。另外，对5株从病猪体内分离的猪胸膜肺炎放线杆菌进行了检测，5株均成阳性反应；对10只屠宰猪的肺脏分离物进行了检测，结果1份为阳性。结果表明此法特异性和敏感性均很高，可做为猪传染性胸膜肺炎的快速诊断和流行病学调查的手段。     

猪传染性胸膜肺炎放线杆菌参考株抗血清的制备及野毒株血清型鉴定

将猪传染性胸膜肺炎放线杆菌全部12个血清型的国际参考菌株接种于培养基大量培养。回收菌体、甲醛灭活后，加入蜂胶佐剂制成疫苗。分别免疫健康的试验用白兔。获得针对该菌单一血清型的抗血清。虽然不同血清型间有一定的交叉反应，但对同源菌株的菌体抗原的凝集效价最高。分别可达6～8个log2。用该套血清对2003年从山东省各地分离到的猪传染性胸膜肺炎放线杆菌野毒株11株进行了血清型鉴定。分别为3型（2株）、4型（1株）、5型（4株）和7型（4株）。