不同LET辐射对健康人血淋巴细胞和肿瘤细胞的损伤效应

目的: 探讨不同LET辐射对健康人外周血淋巴细胞的遗传损伤效应；了解重离子辐射诱导人血淋巴细胞染色体畸变的特点；研究用G2期染色体畸变预测肿瘤细胞辐射敏感性的可行性。材料与方法：采用兰州近代物理研究所重离子研究装置产生的12C离子和兰州大学第一附属医院放疗科提供的X射线照射健康人外周血，用常规染色体分析技术和姊妹染色体差别染色法研究不同LET辐射对健康人血淋巴细胞中期染色体的损伤效应、高LET辐射的剂量率效应、时程效应和染色体畸变的特点；用Calyculin A诱导间期染色体凝集技术研究12C 离子对淋巴细胞G2期染色体的损伤效应。用60Coγ射线照射人卵巢癌细胞和肝癌细胞，以细胞克隆存活率和G2期染色体畸变为生物学终点，探讨用G2期染色体畸变预测肿瘤细胞辐射敏感性的可行性。 结果与结论 1. 不同LET辐射诱导‘双+环’畸变与剂量之间存在良好的线性平方关系， 12C 离子诱导的畸变在细胞间的分布不符合泊松分布； 12C离子的相对生物学效应随着LET的增大而增大；12C离子诱导染色体畸变在1－5 Gy/min的范围内不存在剂量率效应；在0－4 Gy的剂量范围内不存在时程效应，说明培养48小时后中期‘双+环’畸变能很好的反应12C离子对淋巴细胞的损伤效应。本研究可以帮助人们了解重离子的相对生物学效应，为病人健康组织的保护以及放射医师的防护提供重要的理论数据。 2. LET为34.6 keV/μm的12C离子诱导淋巴细胞G2-期染色体数目畸变与剂量之间存在良好的线性关系(r=0.99)；最长G2-PCC与最短G2-PCC的长度之比与剂量之间存在良好的线性平方关系(r2=0.96)。表明有希望用G2-期染色体畸变评估重离子的相对生物学效应和估计重离子的辐射剂量，也为合理评估空间混合辐射场中重离子辐射所占比例提供了两个潜在的指标。 3. LET为34.6 keV/μm12C 离子诱导的畸变，除大部分染色体型畸变外，还出现少量的染色单体畸变，这一现象在理论上突破了传统的认识，对重离子辐射损伤机理的认识有重要意义。这可能是由于重离子特殊的离子径迹结构诱导染色单体畸变。重离子相对生物学效应是相对于常规射线而言的，而常规射线辐照G0期淋巴细胞只产生染色体型畸变，因此这一现象的存在对重离子相对生物学效应的确定提出了新的问题。 4. γ射线诱导肿瘤细胞G2期染色体初始断裂畸变和修复24小时后残余断裂畸变都与辐射剂量有良好的相关性；肿瘤细胞克隆存活率与G2染色单体初始断裂(r=0.96)和修复24小时后残余断裂(r=0.91)都有一定的相关性，比较而言，G2染色单体初始断裂畸变能更好的反映肿瘤细胞的辐射敏感性，预示G2染色单体初始断裂畸变有希望成为肿瘤细胞辐射敏感性的预测指标。 5. 2 Gyγ射线诱导的G2-染色单体断裂畸变，有近65% 的断裂在辐射后24小时内得以修复；对G2等点染色单体断裂畸变，在辐射后24小时内只有20%左右得以修复；两类畸变的修复主要发生在辐射后2小时内。 6. 在用G2-assay法测定染色体畸变的实验中，给予较高剂量时，很难获得足够的中期细胞以供观察。用G2-assay和G2-PCC技术获得的数据都与细胞克隆存活率有一定的相关性，比较而言，后者的相关性要好一些。表明G2-PCC技术可以作为细胞存活实验和用常规染色体畸变分析放射敏感性的替代方法

Capillary electrophoretic study on UDP-sugars in cells

Glucose is an important regulator of cell growth and metabolism. Uridine diphosphate sugars (UDP-sugars), as the intermediate products of metabolism, play pivotal roles as precursors in the synthesis of complex carbohydrates and glycolipids as well as lectose. It is very important to study their metabolism in cells in clinical biochemistry. A capillary electrophoretic method has been developed for the analysis of UDP-sugars and nucleotides, By using an uncoated capillary (70cm x 50 mu m) and 20 mmol/L borax buffer (pH 9), 4 important UDP-sugars can be analyzed in 15 min at 22 kV with satisfactory precision and sensitivity. The developed method has been applied to analyze UDP-sugars concentrations in lymphocytes, fibroblasts and mesangial cells, and the results show it not only is much better than HPLC method, but also can be used to measure the energy charge of cells.

Prediction of C-13-nuclear magnetic resonance chemical shifts for aliphatic amines

Prediction of C-13-nuclear magnetic resonance chemical shifts for aliphatic amines is performed. The topological, geological and electronic descriptors are generated. To reduce the variables, the best subsets of the descriptors are obtained by using leaps-and-bounds regression analysis. The model is achieved using multiple regression with satisfactory results.

Expression of a twist-related gene, Bbtwist, during the development of a lancelet species and its relation to cephalochordate anterior structures

Mesoderm formation plays a crucial role in the establishment of the chordate body plan. In this regard, lancelet embryos develop structures such as the anteriorly extended notochord and the lateral divertecula in their anterior body. To elucidate the developmental basis of these structures, we examined the expression pattern of a lancelet twist-related gene, Bbtwist, from the late gastrula to larval stages. In late-gastrula embryos, the transcripts of Bbtwist were detected in the presumptive first pair of somites and the middorsal wall of the primitive gut. The expression of Bbtwist was then upregulated in the lateral wall of somites and the notochord. At the late-neurula stage, it was also expressed in the anterior wall of the primitive gut, as well as in the evaginating lateral diverticula. No signal was detected in the left lateral diverticulum when it was separated from the gut, while in the right one, the gene was expressed later during the formation of the head coelom in knife-shaped larvae, and in the anterior part of the notochord in the same larvae. In 36-h larvae, only faint expression was detected in the differentiating notochordal and paraxial mesoderm in the caudal region. These expression patterns suggest that Bbtwist is involved in early differentiation of mesodermal subsets as seen in Drosophila and vertebrates. The expression in the anterior notochord may be related to its anterior expansion. The expression in the anterior wall of the primitive gut and its derivative, the lateral diverticula, suggests that lancelets share the capability to produce a mesodermal population from the tip of the primitive gut with nonchordate deuterostome embryos. (C) 1998 Academic Press.

Identification and analysis of a Sciaenops ocellatus ISG15 homologue that is involved in host immune defense against bacterial infection

ISG15 is an interferon-stimulated gene that encodes a ubiquitin-like protein. ISG15 homologues have been identified in a number of fish species, some of which are known to be regulated at expression level by virus infection and lipopolysacchande (LPS) treatment However, the relationship between ISG15 and live bacterial infection has not been investigated in piscine models. In this study, an ISG15 homologue, SoISG15, was identified from red drum Scraeriops ocellaws and analyzed at expression and functional levels The open reading frame ofSolSG15 is 477 base pairs (bp) and mtronless, with a 5'-untranslated region (UTR) of 91 bp and a 3'-UTR of 415 bp The deduced amino acid sequence of S0ISG15 shares 60-67% overall identities with the ISG15 of several fish species. S0ISG15 possesses two conserved ubiquinn-like domains and the canonical ubiquitin conjugation motif, LRGG, at the C-terminus. Expressional analysis showed that constitutive expression of SolSG15 was highest in blood and lowest in kidney Experimental challenges with LPS and bacterial pathogens induced significant S0ISG15 expression in the kidney but not in the liver Similar differential induction was also observed at cellular level with primary hepatocytes and head kidney (HK) lymphocytes. Poly(' C), however, effected drastic induction of S0ISG15 expression in kidney and liver at both tissue and cellular levels. Immunoblot analysis showed that S0ISG15 was secreted by cultured HK lymphocytes into the extracellular milieu. Recombinant S0ISG15 expressed in and purified from Eschenclua colt was able to enhance the respiratory burst activity, acid phosphatase activity, and bactericidal activity of HK macrophages. Taken together, the results of this study indicated that SoISG 15 possesses apparent immunological property and is likely to be involved in host immune defense against bacterial infection. (C)2010 Elsevier Ltd All rights reserved.

Morphological and ultrastructural characterization of the coelomocytes in Apostichopus japonicus

The coelomocytes suspended in the coelomic fluid and occurring in the coelomic epithelial layer of the sea cucumber Apostichopus japonicus (Selenka) (Holothuroidea: Aspidochirota: Stichopodidae) function as mediators of the immune system, trephocytic cells and nutrient transport cells. Types of coelomocytes are characterized based on their morphological and ultrastructural features. Flow cytometry plus light and electron microscopic analyses were conducted in order to characterize the coelomocytes of A. japonicus. Six types of coelomocytes were identified: lymphocytes, morula cells, amoebocytes, crystal cells, fusiform cells and vibratile cells. Within these major categories, several distinctive cell types occurred that might represent developmental stages. The mean +/- SD coelomocyte concentration in the individuals (body length: 10 to 15 cm; weight: 100 to 150 g) was (3.79 +/- 0.65) X 10(6) cells ml(-1). The coelomic fluid contained mainly hyalinocytes (76.69%) and granulocytes (23.31 %).

Effect of ethanol extract of alga Laurencia supplementation on DNA oxidation and alkylation damage in mice

Laurencia terpenoid extract (LET) had been extracted from the red alga Laurencia tristicha. The study is to investigate the effects of LET supplementation on DNA oxidation and alkylation damages in mice. Forty healthy kunming mice weighing between 18g and 25g were randomly assigned into 4 groups, each consisting of ten animals. The mice were orally intubated respectively for 60 days with the designed concentrations of LET (25, 50, 100 mg/kg b.w.) for three exposed groups and salad oil (0.2 ml) for the blank group. Food and water were free for the animals. Mice in the blank and exposed groups were sacrificed after the last treatment and the blood of each animal was quickly taken for further experiments. The spontaneous and oxidized DNA damages of peripheral lymphocytes induced by H2O2 were analysed by SCGE. O-6-Methy-guanine (O-6-MeG) was measured by high performance capillary zone electrophoresis. There was no significantly difference in DNA spontaneous damage on peripheral lymphocytes of all the mice. The oxidative DNA damage in the 50 mg/Kg body weight supplement group are 286AU with the oxidation of 10 mu mol/L H2O2, significantly lower than the blank group 332AU (p<0.05). The contents of O-6-MeG in plasma in the 50mg/kg b.w. and 100mg/kg b.w. supplement group were 1.50 mu mol/L andl.88 mu mol/L, significantly lower than that of the blank group, which was 2.89 mu mol/L(p<0.05). The results from the present study indicated that the LET were rich in terpenoids and safety to be taken orally and it could improve antioxidative and decrease DNA damage effectively.

一个多元逐步回归的二叉树分类器的设计与实现

作者设计并实现了一个基于多变元逐步回归的二叉树分类器.在树结构和特征子集的选择中采用了穷举法,比有限制条件的选择更合理更优化.用 FORTRAN 语言实现的“遍历”二叉树,充分利用了 FORTRAN 处理可调数组的能力,并采取适当技巧,从而最大限度地利用了计算机内存.该通用分类器,可用来对任何具有统计数据的模式进行分类.在对白血球的分类中,取得了五类97%,六类92.2%的高识别率.

心理干预对围手术期乳腺癌患者情绪、自评健康及免疫功能的影响研究

Objective：Developing a generalized psychological intervention program, and explore its influence on the emotion, subjective health, and immunity function of the perioperation patients with breast cancer. Method：Sixty patients with breast cancer were randomly divided into intervention and control groups. The clinical psychological intervention was performed on patients in the intervention group for 20 days, in addition to the routine therapy and care. Levels of emotion (SAS & SDS), subjective health (SF-36), and immunity function (t lymphocyte subsets) of the patients were tested. Results: 1.There was no significant difference between the age, income, educational level, and type of prefession of the two groups. There was no significant difference between SAS, SDS, SF-36 and lymphocyte subsets（CD3+, CD4+, CD8+, CD4+/CD8+, NK） of the two groups. 2. Scores of SAS and SDS decreased significantly after intervention in experimental group, while the score of SF-36, the average value of CD4+, CD4+/CD8+, and NK increased significantly. For the control group, the score of depression decreased significantly after intervention, while the score of PF, GH, VT, SF, RE, and MH increased significantly. 3. In comparison of the intervention and control group, the intervention effect of SAS, SDS, SF-36 scores (except SF), CD3+, CD4+, CD4+/CD8+, and NK differed significantly, with the priority of experimental group. 4. SDS, SAS, and CD3+, CD4+, NK correlated in negative respectively, while SDS, SAS, and CD8+ correlated in positive. PF, RP, GH, SF, and MH of subjective health correlated in positive with every index of immunity function in positive, except negative correlation with CD4+/CD8+. BP, RE correlated with CD3+,CD4+,CD8+, and NK in positive. VT correlated in positive with CD3+, CD8+, and NK, in negative with CD4+/CD8+. Conclusions: 1. Anxiety, depression, and subjective health, correlated with immunity function in perioperation patients with breast cancer. 2. Psychological intervention can improve the emotional status, subjective health, and immune function of patients with breast cancer to the optimum in perioperative period.

条件反射性免疫抑制及其脑机制的c-fos研究

To explore the neural mechanisms underlying conditioned immunomodulation, this study employed the classical taste aversion (CTA) behavioral paradigm to establish the conditioned humoral and cellular immunosuppression (CIS) in Wistar rats, by paring saccharin (CS) with intraperitoneal (i.p.) injection of an immunosuppressive drug cyclophophamide (UCS). C-fos immunohistochemistry method was used to observe the changes of the neuronal activities in the rat brain during the acquisition, expression and extinction of the conditioned immunosuppression (CIS). The followings are the main results: 1. Five days after one trial of CS-UCS paring, reexposure to CS alone significantly decreased the level of the anti-ovalbumin (OVA) IgG in the peripheral serum. Two trials of CS-UCS paring and three reexposures to CS not only resulted in further suppression of the primary immune response, but also reduced the numbers of peripheral lymphocytes and white blood cells. This finding indicates that CS can induce suppression of the immune function, and the magnitude of the effects is dependent on the intensity of training. 2. On day 5 following two trials of CS-UCS pairing, CS suppressed the spleen lymphocytes responsiveness to mitogens ConA, PHA and PWM, and decreased the numbers of peripheral lymphocytes and white blood cells. On day 15, only PHA induced lymphocyte proliferation was suppressed by CS. On day 30, presentation of CS did not have any effect on these immune parameters. These results suggest that the conditioned suppression of the cellular immune function can retain 5-15 days, and extinct after 30 days. 3. CTA was easily induced by one or two CS-UCS parings, and remained robust even after 30 days. These data demonstrate that CIS can be dissociated from CTA, and they may be mediated by different neural mechanisms. 4. Immunohistochemistry assays revealed a broad pattern of c-fos expression throughout the rat brain following the CS-UCS pairing and reexposure to CS, suggesting that many brain regions are involved in CIS. Some brain areas including the solitary tract nucleus (Sol), lateral parabrachial nucleus (LPB) and insular cortex (IC), showed high level c-fos expressions in response to both CS and UCS, suggesting that they may be involved in the transmission and integration of the CS and UCS signals in the brain. There were dense c-FOS positive neurons in the paraverntricular nucleus (PVN) and supraoptic nucleus (SO) of hypothalamus, subfornical organ (SFO) and area postrema (AP) etc. after two trials of CS-UCS paring and after the reexposure to CS 5 days later, but not in the first training and after the extinction of CIS (30 days later). The results reflect that these nuclei may have an important role in CIS expression, and may also response to the immunosuppression of UCS. The conditioned training and reexposure to CS 5 days later induced high level c-fos expression in the cingulate cortex (Cg), central amygdaloid nucleus (Ce), intermediate part of lateral septal nucleus (LSI) and ventrolateral parabrachial nucleus (VLPB) etc. But c-fos induction was not apparent when presenting CS 30 days later. These brain regions are mainly involved in CIS, and may be critical structures in the acquisition and expression of CIS. Some brain regions, including the frontal cortex (Fr), ventral orbital cortex (VO), IC, perirhinal cortex (PRh), LPB and the medial part of solitary nucleus (SolM), showed robust c-FOS expression following the conditioning training and reexposure to CS both on day 5 and day 30, suggesting that they are critically involved in CTA.