Cytotoxic T lymphocyte epitopes from human heparanase can elicit a potent anti-tumor immune response in mice

Heparanase is expressed in almost all advanced tumors, and therefore it may serve as a potential target for tumor therapy. Our previous study has shown that heparanase can serve as a potential universal tumor-associated antigen (TAA) for the immunotherapy of advanced tumors. Further study demonstrated that the HLA-A*0201-restricted Cytotoxic T lymphocytes (CTL) epitopes Hpa525 (PAFSYSFFV), Hpa277 (KMLKSFLKA) and Hpa405 (WLSLLFKKL) from human heparanase could induce a potent anti-tumor immune response in vitro. The present study was designed to investigate whether the above peptides could induce immune responses in mice. Our results demonstrated that the effectors from heparanase peptide-immunized mice could effectively lyse various tumor cells that were heparanase positive and HLA-A*0201 matched. We also found that these peptide-specific CTLs did not lyse autologous lymphocytes that had low heparanase activity. Further study revealed that Hpa525, Hpa277, and Hpa405 peptides increased the frequency of IFN-γ-producing T cells as compared to a negative peptide. These results suggest that Hpa525, Hpa277, and Hpa405 peptides are novel HLA-A*0201-restricted CTL epitopes capable of inducing heparanase-specific CTLs in mice. Because heparanase is expressed in most advanced malignant tumors, Hpa525, Hpa277, and Hpa405 peptide-based vaccines may be useful for the immunotherapy of patients with advanced tumors.     

Effects of Sevoflurane on Self-Renewal Capacity and Differentiation of Cultured Neural Stem Cells

Sevoflurane anesthesia in infant rats can result in long-term cognitive impairment, possibly by inhibiting neurogenesis. The hippocampus is critical for memory consolidation and is one of only two mammalian brain regions where neural stem cells (NSCs) are renewed continuously throughout life. To elucidate the pathogenesis of sevoflurane-induced cognitive dysfunction, we measured the effects of clinical sevoflurane doses on the survival, proliferation, and differentiation of hippocampal NSCs. Neural stem cells were isolated from Sprague–Dawley rat embryos, expanded in vitro, and exposed to sevoflurane at 0.5, 1, or 1.5 minimal alveolar concentration (MAC) for 1 or 6 h. Two days after treatment, cell viability, cytotoxicity, and apoptosis rate were estimated by WST-1 assay, lactate dehydrogenase (LDH) activity, and TdT-mediated dUTP-biotin nick end labeling (TUNEL), respectively, while proliferation rate was assessed by 5-ethynyl-2′-deoxyuridine (BrdU) incorporation and Ki67 staining. Differentiation was assayed 7 days after treatment by immunocytochemistry and Western blots of neuron and glial markers. The phosphorylation level of p44/42 extracellular regulated kinases (ERK1/2) was measured in the proliferation and differentiation phases respectively. Sevoflurane at 1 MAC or 1.5 MAC for 1 h increased viable cell number whereas a 6 h exposure at these same concentrations suppressed proliferation and promoted apoptotic death (P < 0.01). Sevoflurane had no effect on NSC differentiation, and a sub-clinical concentration (0.5 MAC) altered neither proliferation nor viability. The phosphorylation level of ERK1/2 increased after 1 h of 1 MAC or 1.5 MAC of sevoflurane exposure in the proliferation phase, but not in the differentiation phase. Brief (1 h) exposure to sevoflurane at clinical concentrations enhanced proliferation of cultured NSCs possibly mediated by ERK1/2, but a 6 h exposure suppressed proliferation and induced apoptosis. Prolonged sevoflurane exposure may decrease the self-renewal capacity of hippocampal NSCs, resulting in cognitive deficits.     

四川省森林植被固碳经济价值动态

<正>确估算森林植被固碳经济价值可为森林生态系统的生态效益评价提供基础数据。利用1997年和2014年两期四川省森林资源清查数据,依据不同森林类型的生物量与蓄积量回归方程和支付意愿法,估算了四川省两个时期森林植被的固碳经济价值。结果表明,从1997年到2014年,四川省森林植被固碳经济价值由703.17亿元增长到865.75亿元,净增长162.58亿元,年均增长9.56亿元,年均增长率为1.36%;在两个时期,云冷杉林的固碳经济价值比重最大,分别占总固碳经济价值的54.82%和46.62%,表明云冷杉森林植被类型在全省森林植被固碳经济价值中占有重要的地位;四川省天然林和人工林植被的固碳经济价值均呈增加趋势,并且人工林植被固碳经济价值年均增长速率(7.42%)明显高于天然林(1.03%);四川省森林植被固碳经济价值总体上随林龄的增加而增加。研究结果说明,实施包括天然林保护工程在内的森林保护和经营管理措施对提高森林植被的固碳经济价值具有重要的作用。     

三江源牧户参与草地生态保护的意愿

牧民对环境保护的响应是生态保护战略有效实施的关键,主体参与意愿直接影响生态保护项目实施的成效和可持续性,是区域生态经济可持续发展的前提。通过对三江源地区约283户藏族牧民通过翻译进行结构式访谈,采用Logistic模型从主体角度探讨了牧户愿意参与生态保护行为响应的主要影响因素。结果表明:(1)三江源区域约87%的牧户认为生态保护对牧户有好处,但受各种内外部因素的影响只有近70%的牧户是在政府主导下基于有限理性而被动的参与生态保护行为响应。(2)三江源牧户参与生态保护行为响应的意愿主要受当地政府的保护力度及牧户对生态保护外部性的认知水平、生计水平、外界接触程度、工作机会的正影响,并受牧户的年龄、离中心城镇的距离和区域气候恶劣情况等因素的负影响,系数依次为:2.22、3.98、1.93、2.26、1.48、-1.63、-2.43、-0.92。(3)牧户的生计水平、退化感知和外部性认知是影响三江源牧户参与生态保护意愿的关键因素,牧户参与生态保护意愿的概率不仅仅是牧户出于自身利益和未知风险考虑下被动的响应,更是当地政府的环境知识宣传和保护投入影响下个体的抉择结果。(4)创造更多的就业机会和解决牧户的单一化生计问题,构建完善地生态补偿机制让牧户分享生态保护的外部性效益,并激励牧户主动参与生态保护行为响应,才能最终实现区域生态保护、牧户幸福和生态经济可持续发展的多赢局面。     

Benzo(a)pyrene-induced mitochondrial dysfunction and cell death in p53-null Hep3B cells

Benzo(a)pyrene (BaP) has been shown to induce apoptosis and necrosis in various cell types. However, the effect of BaP on mitochondria function and p73, and their possible roles in BaP-induced cell death have not been well studied. This study focused on mitochondria-mediated cell death and the occurrence of p73 protein accumulation in BaP-treated human hepatoma Hep3B (p53-null) cells. We found that BaP (8, 16, 32 and 64μM) induced early necrosis at 12h and delayed apoptosis at 24h. BaP dramatically induced ethoxyresorufin-O-deethylase activity and led to significant increase in oxidative stress at early time points (6 and 12h). Necrotic cell death was concurrent with loss of mitochondrial membrane potential, decrease in the ATP level and activities of Na(+)/K(+)-ATPase and Ca(2+)/Mg(2+)-ATPase. However, these changes were reversed in the process of apoptosis. In addition, after BaP treatment, c-Jun N-terminal kinase (JNK) and Bax were activated during apoptosis and no change in p73 protein level was observed. These results revealed that the cells with mitochondria dysfunction and ATP depletion underwent necrosis at early time point and apoptosis afterward when they recovered from mitochondrial dysfunction and ATP depletion. Activation of JNK and Bax possibly contributed to BaP-induced apoptosis.     

Protein structural plasticity exemplified by insertion and deletion mutants in T4 lysozyme.

To further investigate the ways in which proteins respond to changes in the length of the polypeptide chain, a series of 32 insertions and five deletions were made within nine different alpha-helices of T4 lysozyme. In most cases, the inserted amino acid was a single alanine, although in some instances up to four residues, not necessarily alanine, were used. Different insertions destabilized the protein by different amounts, ranging from approximately 1 to 6 kcal/mol. In one case, no protein could be obtained. An "extension" mutant in which the carboxy terminus of the molecule was extended by four alanines increased stability by 0.3 kcal/mol. For the deletions, the loss in stability ranged from approximately 3 to 5 kcal/mol. The structures of six insertion mutants, as well as one deletion mutant and the extension mutant, were determined, three in crystal forms nonisomorphous with wild type. In all cases, including previously described insertion mutants within a single alpha-helix, there appears to be a strong tendency to preserve the helix by translocating residues so that the effects of the insertion are propagated into a bend or loop at one end or the other of the helix. In three mutants, even the hydrophobic core was disrupted so as to permit the preservation of the alpha-helix containing the insertion. Translocation (or "register shift") was also observed for the deletion mutant, in this case a loop at the end of the helix being shortened. In general, when translocation occurs, the reduction in stability is only moderate, averaging 2.5 kcal/mol. Only in the most extreme cases does "bulging" or "looping-out" occur within the body of an alpha-helix, in which case the destabilization is substantial, averaging 4.9 kcal/mol. Looping-out can occur for insertions close to the end of a helix, in which case the destabilization is less severe, averaging 2.6 kcal/mol. Mutant A73-[AAA] as well as mutants R119-[A] and V131-[A], include shifts in the backbone of 3-6 A, extending over 20 residues or more. As a result, residues 114-142, which form a "cap" on the carboxy-terminal domain, undergo substantial reorganizations such that the interface between this "cap" and the rest of the protein is altered substantially. In the case of mutant A73-[AAA], two nearby alpha-helices, which form a bend of approximately 105 degrees in the wild-type structure, reorganize in the mutant structure to form a single, essentially straight helix. These structural responses to mutation demonstrate the plasticity of protein structures and illustrate ways in which their three-dimensional structures might changes during evolution.     

Genetic determination of osteoporosis: lessons learned from a large genome-wide linkage study

Osteoporosis is a common disease with strong genetic control. We performed an autosomal linkage scan in a large pedigree-based sample of 4,498 subjects for a composite osteoporosis phenotype that combines osteoporotic fracture (OF) and low bone mineral density (BMD). All of the subjects were U.S. Caucasians recruited in the Omaha area of Nebraska. Sex-specific linkage analyses and autosomal imprinting analyses were also conducted. For conventional linkage analyses in the total sample, we identified suggestive linkage on chromosomes 14q32 (LOD = 2.61), 7p14 (LOD = 2.42), and 11q25 (LOD = 2.09). In female subjects a significant linkage signal was detected on chromosome 14q22 (LOD = 3.53) and another two peaks were detected on chromosomes 7p14 (LOD = 3.07) and 9p21 (LOD = 2.29). Suggestive evidence of imprinted loci was found with paternally derived alleles on chromosomes 1q42 (LOD = 2.12) and 9q34 (LOD = 1.88). Some evidence of linkage to maternally derived alleles was found on chromosome 7q22 (LOD = 1.67). Our study provides new clues to osteoporosis genetic research and for the first time suggests that genomic imprinting effects may play a role in the etiology of osteoporosis.     

云雾山铁杆蒿茎叶浸提液对封育草地四种优势植物的化感效应

为了揭示铁杆蒿群落在植被演替中的作用,通过研究铁杆蒿茎叶浸提液对铁杆蒿草地4种优势植物(百里香、大针茅、本氏针茅和赖草)的种子萌发及幼苗生长的干扰,结果表明:高浓度的铁杆蒿浸提液(甲醇浸提液和水浸提液)使得百里香、大针茅、本氏针茅和赖草的种子发芽指数降低,发芽率、芽长和根长低于对照,种子平均发芽时间延长达1.13-2.16 d。而低浓度浸提液(0.005 g/mL)使得百里香的发芽要高于对照11.63%(水浸提液)、15.12%(甲醇浸提液)。在幼苗生长期,铁杆蒿浸提液对4种受试植物幼苗芽和根的生长受到不同程度的抑制,不同浓度的浸提液对植物的化感作用强度不同,随浓度的增加,抑制作用越强,0.1 g/mL相对其他浓度有显著性差异;其中0.005 g/mL浸提液对本氏针茅和赖草的幼苗生长有促进作用,幼根生长高出对照19.00%、16.06%,水浸提液对幼芽促进了2.33%、9.06%,在0.1 g/mL浓度下,本氏针茅或大针茅的生长完全受到抑制,芽长和根长抑制率为100%;同一浓度下的不同浸提液对植物的抑制作用也不同,其中百里香对铁杆蒿浸提液的敏感度是最低的;甲醇浸提液的化感作用要强于水浸提液。在封育过程中,百里香群落向铁杆蒿群落的过渡,铁杆蒿的化感作用是该草地演替的一个重要影响因子。