Canine model of ischemic stroke with permanent middle cerebral artery occlusion: clinical and histopathological findings

The aim of the present study was to assess the clinical and histopathological findings in a canine model of ischemic stroke. Cerebral ischemic stroke was induced by middle cerebral artery occlusion in four healthy beagle dogs using silicone plugs. They showed neurological signs of forebrain dysfunction such as reduced responsiveness, head turning, circling, postural reaction deficits, perceptual deficits, and hemianopsia. These signs gradually regressed within 4 weeks without therapy. On magnetic resonance imaging, T2 hyperintensity and T1 hypointensity were found in the cerebral cortex and basal ganglia. These lesions were well-defined and sharply demarcated from adjacent brain parenchyma with a homogenous appearance. No abnormalities of the cerebrospinal fluid were observed. At necropsy, atrophic and necrotic lesions were observed in the cerebral cortex. The cerebral cortex, basal ganglia, and thalamus were partially unstained with triphenyl-tetrazolium chloride. Histopathologically, typical features of infarction were identified in cortical and thalamic lesions. This study demonstrates that our canine model resembles the conditions of real stroke patients.     

Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

We previously induced protective immune response by oral immunization with yeast expressing the ApxIIA antigen. The ApxI antigen is also an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01). The highest systemic (IgG) and local (IgA) humoral immune responses to ApxIA and ApxIIA were detected in group E after the third immunization (p < 0.05). The levels of IL-1β and IL-6 after challenge with an A. pleuropneumoniae field isolate did not change significantly in the vaccinated groups. The level of TNF-α increased in a time-dependent manner in group E but was not significantly different after the challenge. After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05). The survival rate in each group was closely correlated to the immune response and histopathological observations in the lung following the challenge. These results suggested that immunity to the ApxIA antigen is required for optimal protection.     

纳米二氧化钛对纤维素酶活力的影响

作者主要研究不同浓度纳米二氧化钛在不同作用时间、不同温度及不同光照处理条件下对纤维素酶酶活力的影响，以探讨纳米二氧化钛在生物大分子水平上的作用及其作用机理。结果表明：纳米二氧化钛在可见光与紫外光照射条件下均可使纤维素酶失活。而且随着与纤维素酶作用时间的延长、纳米二氧化钛浓度增加和作用温度的升高，其对纤维素酶活力的影响也随之增大。     

穴播条件下株行距对紫花苜蓿种子产量和质量的影响

在甘肃省酒泉市研究了穴播条件下不同株、行距处理对紫花苜蓿种子生产性能的影响，结果表明：在产种第一年（2004年）行距和株距对种子产量和产量构成因素均产生了显著的影响（P〈0．05），随着行距从60cm递增至100cm，种子产量也随之从1240kg／hm^2下降到982kg／hm^2，而各株距处理的种子产量则从15cm的1426kg／hm^2依次递减至60cm的807kg／hm^2；行距60cm和株距15cm组合的产量最高，为1833kg／hm^2，100cm行距和株距60cm组合的产量最低，为753kg／hm^2。株行距的增加显著降低了枝条密度，但结荚花序数／枝条和荚果数／结荚花序却显著提高。品种间种子产量的差异不显著。品种间的结荚花序数／枝条、种子重量、正常种苗率、发芽势和硬实率存在显著的差异（P〈0．05）。     

茶花鸡种群遗传结构分析及其保种措施的探讨

利用14个微卫星标记对云南茶花鸡12个家系的等位基因频率、平均基因杂合度、平均多态信息含量以及家系间的亲缘关系进行了分析。结果表明:共检测到99个等位基因,每个位点的等位基因分布并不均匀,再对12个家系进行分析,将其聚为3类,一类为Z-1,Z-2,Z-3,Z-4,Z-5,Z-6,B-1,一类为Z-7,Z-8,Z-9,Z-10,矮脚系B-2自成一类,这表明了家系间的遗传相似性,同时我们对特殊的矮脚系B-2进行了相关的探讨,最后我们提出了对茶花鸡保种的相关建议。     

猪传染性胃肠炎病毒S与M基因融合表达载体及IRES连接双基因共表达载体的构建

目的:研究如何提高猪传染性胃肠炎病毒(TGEV)中主要抗原基因S与M基因对猪传染性胃肠炎的免疫调节作用,进一步探究基因疫苗的有效开发途径。方法:通过两种方法应用牛β酪蛋白启动子构建S和M融合基因表达载体以及构建IRES连接M和S双基因共表达核酸疫苗载体。结论:成功构建好M和S融合基因表达载体及IRES连接的M和S双基因真核共表达载体,并为利用乳腺生物反应器生产可食性疫苗提供有效措施。     

Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 10⁷ cfu ml⁻¹ (OD_600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation.     

Effects of Sini decoction on the expression of TGF-β1 in balloon injured iliac artery of rabbits

AIM:To investigate the effect of Sini decoction (SND) on vascular stenosis and the expression of transfoming growth factor-β1 (TGF-β₁) in iliac artery balloon injured rabbits. METHODS:24 male New Zealand albino rabbits were divided into three groups:control group, model group and SND treatment group. The iliac arteries were injured by balloon in model and SND groups. Four weeks later, serum TGF-β₁ level was assayed by ELISA. Endothelial hyperplasia, TGF-β₁ protein and mRNA expression were observed in injured iliac artery. RESULTS:Light microscope showed that the vascular lumina were narrower, intima was thicker in model group control and SND treatment group. The serum TGF-β₁ level was lower in control than model group and SND treatment group, and the serum TGF-β₁ level in SND treatment group was lower than that in model group. Immunohistochemistry and RT-PCR results showed that TGF-β₁ protein and mRNA expression was lower in rabbit iliac artery of control group than that in model group and SND treatment group, and the expression of TGF-β₁ protein and mRNA decreased significantly in SND treatment group compared with model group. CONCLUSION:SND could lessen intimal hyperplasia and vascular stenosis in balloon injured iliac artery, which might be related to decrease in TGF-β₁ protein and gene expression in iliac artery.      

14-3-3σ negatively regulates Akt and inhibits Rat1-Akt cell xenografts

AIM:The present study was designed to investigate the effect of adenovirus-mediated 14-3-3σ on Rat1-Akt cell xenografts and to explore whether the effect was mediated through negative regulation of Akt. METHODS:The effect of Ad-14-3-3σ on Rat1-Akt cells xenografts was observed in nude mice ex vivo and in vivo. Western blotting was used to detect 14-3-3σ protein,phospho-Akt (Thr 308), phospho-Akt (Ser 473), and phospho-(Ser/Thr) Akt substrate in tumor tissue after transfection of 14-3-3σ gene. RESULTS:The tumor volume was dramatically decreased and its emergence was delayed, regardless of using Ad-14-3-3σ-treated Rat1-Akt cells (ex vivo) or injected Ad-14-3-3σ in vivo, of which the effect of the continuously injected group was the best. Levels of Akt protein, phosphorylated Akt and phosphorylated Akt substrates in tumors obtained from Ad-14-3-3σ-treated mice were markedly less than those in PBS or Ad-β-gal-treated mice. CONCLUSION:14-3-3σ suppressed Akt overexpession in cells of Rat1-Akt xenografts by negatively regulating Akt.     

Relationship between uPA and NF-κB p65 expression and its clin ical significance in breast cancer

AIM:To evaluate the relationship between u PA and NF-κB p65 expression and its clinical significance in breast cancer.METHODS:The uPA mRNA was measured by real-time fluorescent quan t itative PCR in 46 cases of breast cancer tissues and their adjacent counterparts .NF-κB p65 were measured using immunohistochemistry.RESULTS:The expression of uPA gene was elevated in 63% of cases ,and there was a strong correlation between NF-κB p65 and uPA expression (r ＝0.451，P<0.01).In addition,the expression of uPA was associated with num bers of axillary lymph node involvement and tumor size.However,no significance was found in uPA expression in the age,pathological type and distant metastasi s organ classification.CONCLUSION:The uPA gene is highly expressed in most of the brea st cancer,and there is strong correlation to NF-κB p65 expression,number of l ymph nodes involvement and tumor size.