钩端螺旋体外膜疫苗的流行病学效果评价

为了评价国产钩端螺旋体外膜疫苗的流行病学效果。于1998年6－10月,用队列研究和病例对照研究等方法在钩体病流行区5－60岁农业人口 ,观察比较接种组和对照组钩体发病情况。队列研究显示,钩体外膜疫苗对同血清群钩体的保护率为75．17％,效果指烽为4．03。1：2配对调查的疫苗保护率为81．25％,1：3配对结果为73．33％。用筛选法估计的疫苗效果为755。不同研究方法的结果相近,均一致说明,该疫苗的流行病学效果较理想。研究结果还显示,该疫苗对异血清群钩体也有一定的保护作用。     

川中丘陵地区大豆根瘤菌遗传多样性与系统发育关系

【目的】研究分离自川中丘陵地区大豆根瘤菌的遗传多样性和系统发育。【方法】采用16S rDNA PCR-RFLP和16S rRNA基因、glnII、共生基因(nodC)系统发育分析的方法进行研究。【结果】供试未知菌的16S rDNA用4种限制性内切酶(HaeⅢ、HinfⅠ、MspⅠ及TaqⅠ)酶切后获得5种16S遗传图谱类型。16S rDNA PCR-RFLP结果表明,所有供试菌株在83%水平分为慢生根瘤菌属(Bradyrhizobium)和中华根瘤菌属(Sinonrhizobium)两大类群,而75%的菌株为中华根瘤菌。6个代表菌株的16S rDNA、glnII和nodC三个位点基因的系统发育结果基本一致,4株与S.fredii USDA205T相似度最高;有2株分别与B.yuanmingense CCBAU10071T、B.diazoefficiens USDA110T相似度最高。4个Sinonrhizobium代表菌株16S rDNA、glnII序列相似度分别为98.3%-99.9%、98.2%-100%,但它们的nodC基因序列完全相同。【结论】川中丘陵地区大豆根瘤菌具有较丰富的遗传多样性,S.fredii为优势种。     

Enhanced Production of Ganoderic Acids and Cytotoxicity of Ganoderma lucidum Using Solid-Medium Culture

Submerged cultures of Ganoderma lucidum are used to produce fungal mycelium, which is used as a functional food and in the production of various triterpenoids, including ganoderic acids (GAs). Specific culture approaches that produce fungal mycelium with high levels of GAs and good biological activity are critical in the functional food industry. In this study, a solid-medium culture approach to producing mycelium was compared to the submerged culture system. Production of GAs, biomass, intracellular polysaccharides, and cytotoxicity of the cultured mycelium were compared as between solid and submerged culture. Growing G. lucidum strains on solid potato dextrose agar medium increased biomass, the production of ganoderic acid 24 (lanosta-7,9(11), 24-trien-3α-o1-26-oic acid), GAs, and total intracellular polysaccharides as compared to fungi grown in submerged culture. Triterpenoid-enriched methanol extracts of mycelium from solid-medium culture showed higher cytotoxicity than those from submerged culture. The IC(50) values of methanol extracts from solid-medium culture were 11.5, 8.6, and 9.9 times less than submerged culture on human lung cancer cells CH27, melanoma cells M21, and oral cancer cells HSC-3 respectively. The squalene synthase and lanosterol synthase coding genes had higher expression on the culture of solid potato dextrose medium. This is the first report that solid-medium culture is able to increase GA production significantly as compared to submerged culture and, in the process, produces much higher biological activity. This indicates that it may be possible to enhance the production of GAs by implementing mycelium culture on solid medium.     

云南松三种同域共存切梢小蠹梢转干期的空间分布格局

分布在我国西南地区的横坑切梢小蠹,云南切梢小蠹和短毛切梢小蠹同域危害寄主云南松,给林业生产带来巨大损失。为探讨同域切梢小蠹种群在共存下对其空间分布格局的影响,采用传统聚集指标法和地统计学方法研究了三者在梢转干期不同受害云南松纯林树冠中的空间分布型。结果表明重度受害样地中云南切梢小蠹种群密度显著高于横坑切梢小蠹,在轻度受害样地则相反;传统聚集指标法结果显示同域共存的3种切梢小蠹种群在不同受害程度云南松中均为聚集分布,横坑切梢小蠹和云南切梢小蠹聚集是由环境因素和昆虫本身的聚集习性引起;地统计学结果表明除重度受害样地中短毛切梢小蠹呈随机分布外,其余切梢小蠹在不同种群密度下均呈聚集分布;除重度受害样地横坑切梢小蠹外,其他小蠹的空间依赖范围为4.01—7.45 m。横坑切梢小蠹和云南切梢小蠹在不同受害林分中拟合的半变异函数模型在球形模型和高斯模型之间转换。同域共存关系不影响不同种群密度下的切梢小蠹种群空间分布类型,但影响其半变异函数模型和理论参数。     

两种海洋赤潮微藻赤潮异弯藻和米氏凯伦藻之间的相互作用

在实验生态条件下研究了不同起始生物量比的两种海洋赤潮微藻赤潮异弯藻(Heterosigma akashiwo)和米氏凯伦藻(Karenia mikimotoi)的种群增长特征。结果发现: 1)在单培养体系中, H. akashiwo和K. mikimotoi的种群增长均可用逻辑斯谛增长模型(Logistic equation)拟合, 但不同的起始密度比对两种微藻的生长可产生显著影响: 随着起始密度的增加, 种群的瞬时增长率(r)随之增加, 但环境负载能力(K)逐渐降低, 进入指数增长期和静止期的时间也相应缩短。2)在共培养体系中, 两种微藻的K值都受到明显的抑制, 与对照组(单培养体系)相比差异显著(p＜0.05); 不同起始生物量比对共培养体系中两种微藻的生长和竞争影响显著: 当H. akashiwo和K. mikimotoi的起始生物量比(H:K)为1:4和1:16时, K. mikimotoi在竞争中占据优势地位; 当H:K=1:1时, H. akashiwo在竞争中占绝对优势。他感作用是导致本实验结果的可能原因。     

彼得森黄群藻孢囊的形成及形态

利用光镜及扫描电镜对彼得森黄群藻的形态结构、孢囊形成进行详细观察和描述.结果显示,彼得森黄群藻分布广泛,营养体形态变化也大,可以分为不同的变种和变型,其孢囊形态也有多种形态.根据鳞片结构,鉴定出其中的2个变型,并对其所产生的孢囊形态进行了对应.     

降低mRNA差异显示技术假阳性率的一种方法

为了探讨降低mRNA差异显示技术假阳性率的方法 ,进一步提高此技术的可靠性 ,提取了手术切除肝癌及非癌肝组织成对标本的总RNA ,逆转录获得cDNA片段 ,以mRNA差异显示方法筛选差异表达基因 ,选取较明显的一条差异表达条带 ,行进一步PCR扩增 .分别对PCR产物及其经TA克隆后随机挑选的 6个单克隆质粒DNA进行序列分析 ,并通过GenBank BLAST数据库进行序列的同源性比较 ,以Northern杂交予以来源确认 .自 72 0余条扩增条带中共选出 2 8条差异条带 .序列分析及同源性比较表明 ,所选择条带的PCR产物为一可能的新基因片段 ;而随机选择的 6个TA克隆质粒DNA中 ,有 4个为同一已知基因片段 ,一个为另一已知基因片段 ,一个为一可能的新基因片段 .同源性比较表明 ,PCR产物直接测序所得序列与TA克隆质粒DNA的 6个片段不具同源性 .结果表明 ,mRNA差异显示条带可能由 1条以上分子量相似的片段构成 ,直接对PCR产物行序列分析并以其为探针进行Northern杂交 ,是导致出现假阳性片段的原因之一 .将PCR产物进行TA克隆 ,对单克隆质粒DNA进行序列分析并以其为探针进行Northern杂交 ,可能是解决此问题的一种较好方法 .     

In Vitro Activity of Sodium New Houttuyfonate Alone and in Combination with Oxacillin or Netilmicin against Methicillin-Resistant Staphylococcus aureus

Background

Staphylococcus aureus can cause severe infections, including bacteremia and sepsis. The spread of methicillin-resistant Staphylococcus aureus (MRSA) highlights the need for novel treatment options. Sodium new houttuyfonate (SNH) is an analogue of houttuynin, the main antibacterial ingredient of Houttuynia cordata Thunb. The aim of this study was to evaluate in vitro activity of SNH and its potential for synergy with antibiotics against hospital-associated MRSA.

Methodology

A total of 103 MRSA clinical isolates recovered in two hospitals in Beijing were evaluated for susceptibility to SNH, oxacillin, cephalothin, meropenem, vancomycin, levofloxacin, minocycline, netilmicin, and trimethoprim/sulfamethoxazole by broth microdilution. Ten isolates were evaluated for potential for synergy between SNH and the antibiotics above by checkerboard assay. Time-kill analysis was performed in three isolates to characterize the kill kinetics of SNH alone and in combination with the antibiotics that engendered synergy in checkerboard assays. Besides, two reference strains were included in all assays.

Principal Findings

SNH inhibited all test strains with minimum inhibitory concentrations (MICs) ranging from 16 to 64 µg/mL in susceptibility tests, and displayed inhibition to bacterial growth in concentration-dependent manner in time-kill analysis. In synergy studies, the combinations of SNH-oxacillin, SNH-cephalothin, SNH-meropenem and SNH-netilmicin showed synergistic effects against 12 MRSA strains with median fractional inhibitory concentration (FIC) indices of 0.38, 0.38, 0.25 and 0.38 in checkerboard assays. In time-kill analysis, SNH at 1/2 MIC in combination with oxacillin at 1/128 to 1/64 MIC or netilmicin at 1/8 to 1/2 MIC decreased the viable colonies by ≥2log₁₀ CFU/mL.

Conclusions/Significance

SNH demonstrated in vitro antibacterial activity against 103 hospital-associated MRSA isolates. Combinations of sub-MIC levels of SNH and oxacillin or netilmicin significantly improved the in vitro antibacterial activity against MRSA compared with either drug alone. The SNH-based combinations showed promise in combating MRSA.     

Lysine-Specific Demethylase 1 in Breast Cancer Cells Contributes to the Production of Endogenous Formaldehyde in the Metastatic Bone Cancer Pain Model of Rats

Background

Bone cancer pain seriously affects the quality of life of cancer patients. Our previous study found that endogenous formaldehyde was produced by cancer cells metastasized into bone marrows and played an important role in bone cancer pain. However, the mechanism of production of this endogenous formaldehyde by metastatic cancer cells was unknown in bone cancer pain rats. Lysine-specific demethylase 1 (LSD1) is one of the major enzymes catalyzing the production of formaldehyde. The expression of LSD1 and the concentration of formaldehyde were up-regulated in many high-risk tumors.

Objective

This study aimed to investigate whether LSD1 in metastasized MRMT-1 breast cancer cells in bone marrows participated in the production of endogenous formaldehyde in bone cancer pain rats.

Methodology/Principal Findings

Concentration of the endogenous formaldehyde was measured by high performance liquid chromatography (HPLC). Endogenous formaldehyde dramatically increased in cultured MRMT-1 breast cancer cells in vitro, in bone marrows and sera of bone cancer pain rats, in tumor tissues and sera of MRMT-1 subcutaneous vaccination model rats in vivo. Formaldehyde at a concentration as low as the above measured (3 mM) induced pain behaviors in normal rats. The expression of LSD1 which mainly located in nuclei of cancer cells significantly increased in bone marrows of bone cancer pain rats from 14 d to 21 d after inoculation. Furthermore, inhibition of LSD1 decreased the production of formaldehyde in MRMT-1 cells in vitro. Intraperitoneal injection of LSD1 inhibitor pargyline from 3 d to 14 d after inoculation of MRMT-1 cancer cells reduced bone cancer pain behaviors.

Conclusion

Our data in the present study, combing our previous report, suggested that in the endogenous formaldehyde-induced pain in bone cancer pain rats, LSD1 in metastasized cancer cells contributed to the production of the endogenous formaldehyde.