骨特异性转录因子Runx2对抗空间骨丢失效应的初步研究

为了构建稳定过表达Runx2 (骨特异性转录因子)的C2C12 (小鼠成肌细胞)和MG63 (前成骨细胞)细胞株, 并用于研究Runx2在对抗空间骨丢失效应中的作用. 利用实时定量聚合酶链式反应鉴定Runx2下游基因I型胶原、碱性磷酸酶表达情况, 在二维回转器中培养稳定细胞株, 通过定量聚合酶链式反应,观察在模拟失重效应下Runx2基因对其下游基因表达的影响. 结果表明, 通过筛选获得稳定转染的C2C12-Runx2和MG63-Runx2细胞株, 经鉴定都能过表达Runx2. 转染后的细胞 I 型胶原和碱性磷酸酶mRNA表达增高. 回转组与对照组相比, MG63, C2C12-Runx2, MG63-Runx2细胞的I型胶原和碱性磷酸酶mRNA表达降低, 但在模拟失重效应下, 转染细胞中I型胶原和碱性磷酸酶的mRNA表达下降程度明显低于未转染细胞株. 所构建的C2C12-Runx2和MG63-Runx2细胞株比较稳定, 并证实Runx2能部分对抗失重引起的成骨特异性分子的降低.      

Galileo/GPS载波相位组合观测值可匹配的研究

详细讨论了Galileo系统的4种载波与GPS L2载波的相位组合观测值的可匹配问题.在模糊度保持为整数的前提下,给出了Galileo/GPS载波相位组合观测值的定义,并对包括系统噪声和观测噪声在内的有关误差影响加以分析,最后给出了组合观测值可匹配的定义和判断可匹配的充要条件.      

辐照和EO灭菌对SIS材料免疫原性的对比研究

免疫反应大小是决定可植入生物材料能否开展临床应用的关键因素之一。研究评估了辐照和环氧乙烷（EO）两种灭菌方式处理后的小肠黏膜下层（SIS）脱细胞外基质材料在体内的免疫反应，旨在为其临床试验的开展提供可行依据。将两种灭菌方式处理的SIS脱细胞外基质材料皮下植入到BALB/c小鼠，第14和28天取样后，系统性地评估了其免疫反应。与仅手术不植入材料的阴性对照组相比，免疫器官（脾和淋巴结）的形态、质量、细胞数、淋巴细胞的体外增殖及酶联免疫吸附检测结果均无显著性差异，证明两种灭菌方式处理后的SIS脱细胞外基质材料都不会对小鼠引起明显的免疫排斥反应。流式细胞术分析及局部H&E染色结果表明，经EO灭菌处理的SIS脱细胞外基质材料对小鼠的免疫刺激更小，是更适用于此材料的灭菌方法。研究结果为SIS脱细胞外基质材料的灭菌程序及其临床应用提供了支持。      

-6°卧床模拟失重对T淋巴细胞的影响及内分泌调节机制的研究

本文研究了-6°卧床模拟失重对T淋巴细胞受到有丝分裂原刺激后增殖能力和外周血总T淋巴细胞亚群(CD3+T细胞)、辅助/炎性T细胞亚群(CD4+T细胞)、杀伤T细胞亚群(CD8+T细胞)、及表达CD25分子细胞数的影响,同时观察生长激素、促肾上腺皮质激素、皮质激素的改变,来探讨免疫功能的变化与内分泌系统改变的关系.结果表明,模拟失重造成的免疫功能下降与内分泌系统紊乱有关.      

星际高速公路

最近，NASA喷气推进实验室的工程师马丁·罗设计出一种盘绕于太阳系各行星间的“高速公路，它类似于一个庞大、曲折的虚拟隧道和管道系统。利用这一系统可大大减少未来太空飞行任务中的燃料消耗量。马丁·罗推算出这种被称为“星际高速公路”的太空飞行系统，并利用这一理论设计出NASA“杰内西斯”号太空船的飞行路径。杰内西斯号目前正行驶在这一“太空高速公路”上，它的任务是采集太阳风粒子并返回地球。根据事先的安排，在大多数宇航任务中，当太空船飞经某一天体如行星或卫星时，它们都会利用天体的引力机制，以便尽量提高飞行速度…     

北极69°N和78°N空间碎片凝视探测的对比分析

利用北极69°N和78°N两套非相干散射雷达的首次空间碎片联合观测数据进行空间碎片参数（距离、速度、散射截面积、等效直径等）的对比分析,得出以下结论:两部雷达探测的碎片高度均主要分布在500～1100km和1400～1600km区间,但78°N雷达探测的碎片数量较多;空间碎片的径向速度均在-1.5～1.5km…-1区间,其中大部分为负值,说明在此次探测试验中碎片运动方向主要以远离雷达或地球为主;ESR雷达探测的空间碎片射截面积约为10-5～10-2m2,等效直径主要分布在4～10cm,而UHF雷达探测的空间碎片散射截面积约为10-6～10-2m2,等效直径主要分布在2～6cm,说明在同一高度上69°N雷达探测能力更强;经合理设置判据参数后得出重复检测次数,78°N雷达和69°N雷达分别有32次和14次重复检测,两部雷达共有4次重复检测.这些结果为空间碎片检测和建模提供了参考.      

模拟微重力对人骨髓间充质干细胞向成骨细胞分化中细胞信号通路影响的分析

通过模拟来研究微重力对hMSC向成骨细胞分化的影响,并利用相关信号通路的激活剂或抑制剂来调节这一分化过程.研究结果表明,在成骨细胞分化诱导条件下,微重力降低了hMSC向成骨细胞定向分化的能力,并且成骨细胞标记性基因的表达明显降低, Runt相关转录因子2(Ruax2)的表达受到抑制.相反,过氧化物酶体增殖激活受体γ(PPARγ2)的表达则增加.同时,微重力也降低了ERK的磷酸化水平,而增加了p38MAPK的磷酸化水平.使用p38MAPK的抑制剂SB203580能够抑制p38MAPK的磷酸化,但不能降低PPARγ2的表达水平.骨形态发生蛋白(BMP)能增加Runx2的表达水平.成纤维细胞生长因子2(FGF2)增加了ERK的磷酸化水平,但也不能显著增加成骨细胞标记性基因的表达水平.采用BMP,FGF2和SB203580三种因子组合来调控微重力下的成骨细胞分化能力,结果表明三者的协同作用能显著逆转微重力对成骨细胞定向分化的生物学效应.研究结果还说明,模拟微重力应该是通过不同的细胞信号通路来抑制成骨细胞分化和提升脂肪细胞分化的.      

基于U-net的紫外极光观测极光卵形态提取

极光卵形态提取是极光研究的重要手段.如何提高强干扰背景下的紫外极光图像极光卵形态提取精度,目前仍是一个难题.本文提出一种基于深度学习语义分割模型U-net的方法,实现了对极光卵形态的高精度提取.在Polar卫星紫外极光观测数据的实验结果表明,该方法相比于已有算法精度更高,对完整型极光卵和缺口型极光卵图像均能得到更加精确的提取结果,特别是针对强日辉干扰、灰度不均匀和对比度低情况下的紫外极光图像时,该方法显示了明显优势.      

二维扰动对下游圆盘压力分布的影响

通过测量阻力和表面压力,研究了干扰圆柱尾迹产生的二维扰动对下游圆盘气动特性的影响,得到圆盘迎风面压力分布的变化规律,并结合以往流场结构的研究,对其机理进行了分析.结果表明在圆盘与干扰圆柱间距比L/D较小时,圆盘迎风面低压区宽度随间距增大迅速增加;当L/D>0.7时,对于同一直径的干扰圆柱,圆盘迎风面低压区宽度基本保持不变;对于不同直径的干扰圆柱,在圆盘前产生低压区的宽度随圆柱直径的增加而变大.此外,利用这种干扰方法,可以降低圆盘阻力,实验得到圆盘阻力可减少约9%,相应的圆柱/圆盘系统的阻力可减小4%.      

气泡雾化喷嘴水平喷射的雾化特性研究

研究了水平喷射、端面注气方式的气泡雾化喷嘴结构及工作参数对雾化及流场特性的影响.试验全部在常温常压下进行,液体采用水,雾化气为压缩空气.用2D PDA测量了液雾的平均直径尺寸分布和速度分布.液体喷射压力变化范围200~600 kPa,气液比变化范围4%~10%左右.试验研究了端面注气方式下,注气孔孔径及数目、混合管长度、液体流动状态及工作参数对雾化特性的影响.结果表明,水平喷射的气泡喷嘴,注气孔的尺寸及数目均对雾化特性产生影响;混合段存在一最佳值范围,在此范围内,喷嘴可获得高的雾化质量;液体旋转流动对雾化特性无显著影响,但可影响两相流动中气泡的分布.     

Effects of space flight exposure on cell growth,tumorigenicity and gene expression in cancer cells

It is well recognized that harsh outer space environment, consisting of microgravity and radiation, poses significant health risks for human cells. To investigate potential effects of the space environment exposure on cancer cells we examined the biological changes in Caski cells carried by the “Shen Zhou IV” spaceship. After exposure for 7 days in spaceflight, 1440 survival subclonal cell lines were established and 4 cell lines were screened. 44F10 and 17E3 were selected because of their increased cell proliferation and tumorigenesis, while 48A9 and 31F2 had slower cytological events. Experiments with cell proliferation assay, flow cytometry, soft agar assay, tumorigenesis assay and DNA microarray analysis have shown that selected cell lines presented multiple biological changes in cell morphology, cell growth, tumorigenicity and gene expression. These results suggest that space environment exposure can make significant biological impact on cancer cells and provide an entry point to find the immunological target of tumorigenesis.     

Expression of p53-regulated genes in human cultured lymphoblastoid TSCE5 and WTK1 cell lines after spaceflight in a frozen state

The 53 kDa tumor suppressor protein p53 is generally thought to contribute to the genetic stability of cells and to protect cells from DNA damage through the activity of p53-centered signal transduction pathways. To clarify the effect of space radiation on the expression of p53-dependent regulated genes, gene expression profiles were compared between two human cultured lymphoblastoid cell lines: one line (TSCE5) has a wild-type p53 gene status, and the other line (WTK1) has a mutated p53 gene status. Frozen human lymphoblastoid cells were stored in a freezer in the International Space Station (ISS) for 133 days. Gene expression was analyzed using DNA chips after culturing the space samples for 6 h on the ground after their return from space. Ground control samples were also cultured for 6 h after being stored in a frozen state on the ground for the same time period that the frozen cells were in space. p53-Dependent gene expression was calculated from the ratio of the gene expression values in wild-type p53 cells and in mutated p53 cells. The expression of 50 p53-dependent genes was up-regulated, and the expression of 94 p53-dependent genes was down-regulated after spaceflight. These expression data identified genes which could be useful in advancing studies in basic space radiation biology. The biological meaning of these results is discussed from the aspect of gene functions in the up- and down-regulated genes after exposure to low doses of space radiation.     

Production and action of cytokines in space.

B6MP102 cells, a continuously cultured murine bone marrow macrophage cell line, were tested for secretion of tumor necrosis factor-alpha and Interleukin-1 during space flight. We found that B6MP102 cells secreted more tumor necrosis factor-alpha and interleukin-1 when stimulated in space with lipopolysaccharide than controls similarly stimulated on earth. This compared to increased secretion of interferon-beta and -gamma by lymphocytes that was measured on the same shuttle flights. Although space flight enhanced B6MP102 secretion of tumor necrosis factor-alpha, an experiment on a subsequent space flight (STS-50) found that cellular cytotoxicity, mediated by tumor necrosis factor-alpha, was inhibited.     

Tumor suppressor function of Betaig-h3 gene in radiation carcinogenesis.

Interaction between cell and extracellular matrix (ECM) plays a crucial role in tumor invasiveness and metastasis. Using an immortalized human bronchial epithelial (BEP2D) cell model, we showed previously that expression of a list of genes including Betaig-h3 (induced by transforming growth factor-beta), DCC (deleted in colorectal cancer), p21(cipl), c-fos, Heat shock protein (HSP27) and cytokeratin 14 were differentially expressed in several independently generated, radiation-induced tumor cell lines (TL1-TL5) relative to parental BEP2D cells. Our previous data further demonstrated that loss of tumor suppressor gene(s) as a likely mechanism of radiation carcinogenesis. In the present study, we chose Betaig-h3 and DCC that were downregulated in tumorigenic cells for further study. Restored expression of Betaig-h3 gene, not DCC gene, by transfecting cDNA into tumor cells resulted in a significant reduction in tumor growth. While integrin receptor alpha 5 beta 1 was overexpressed in tumor cells, its expression was corrected to the level found in control BEP2D cells after Betaig-h3 transfection. These data suggest that Betaig-h3 gene is involved in tumor progression by regulating integrin alpha 5 beta 1 receptor. Furthermore, exogenous TGF- beta 1 induced expression of Betaig-h3 gene and inhibited the growth of both control and tumorigenic BEP2D cells. Therefore, downregulation of Betaig-h3 gene may results from the decreased expression of upstream mediators such as TGF-beta. The findings provide strong evidence that the Betaig-h3 gene has tumor suppressor function in radiation-induced tumorigenic human bronchial epithelial cells and suggest a potential target for interventional therapy.     

Protein expression profile changes in human fibroblasts induced by low dose energetic protons

Extrapolation of known radiation risks to the risks from low dose and low dose-rate exposures of human population, especially prolonged exposures of astronauts in the space radiation environment, relies in part on the mechanistic understanding of radiation induced biological consequences at the molecular level. While some genomic data at the mRNA level are available for cells or animals exposed to radiation, the data at the protein level are still lacking. Here, we studied protein expression profile changes using Panorama antibody microarray chips that contain antibodies to 224 proteins (or their phosphorylated forms) involved in cell signaling that included mostly apoptosis, cytoskeleton, cell cycle and signal transduction. Normal human fibroblasts were cultured until fully confluent and then exposed to 2 cGy of 150 MeV protons at high-dose rate. The proteins were isolated at 2 or 6 h after exposure and labeled with Cy3 for the irradiated cells and with Cy5 for the control samples before loading onto the protein microarray chips. The intensities of the protein spots were analyzed using ScanAlyze software and normalized by the summed fluorescence intensities and the housekeeping proteins. The results showed that low dose protons altered the expression of more than 10% of the proteins listed in the microarray analysis in various protein functional groups. Cell cycle (24%) related proteins were induced by protons and most of them were regulators of G1/S-transition phase. Comparison of the overall protein expression profiles, cell cycle related proteins, cytoskeleton and signal transduction protein groups showed significantly more changes induced by protons compared with other protein functional groups.     

Early effects of low dose C ion or X-ray irradiation on human peripheral blood lymphocytes

The aim of this study was to estimate the acute effects of low dose ¹²C⁶⁺ ions or X-ray radiation on human immune function. The human peripheral blood lymphocytes (HPBL) of seven healthy donors were exposed to 0.05 Gy ¹²C⁶⁺ ions or X-ray radiation and cell responses were measured at 24 h after exposure. The cytotoxic activities of HPBL were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT); the percentages of T and NK cells subsets were detected by flow cytometry; mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real time quantitative RT-PCR (qRT-PCR); and these cytokines protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA). The results showed that the cytotoxic activity of HPBL, mRNA expression of IL-2, IFN-γ and TNF-α in HPBL and their protein levels in supernatant were significantly increased at 24 h after exposure to 0.05 Gy ¹²C⁶⁺ ions radiation and the effects were stronger than observed for X-ray exposure. However, there was no significant change in the percentage of T and NK cells subsets of HPBL. These results suggested that 0.05 Gy high linear energy transfer (LET) ¹²C⁶⁺ radiation was a more effective approach to host immune enhancement than that of low LET X-ray. We conclude that cytokines production might be used as sensitive indicators of acute response to LDI.     

Residual chromatin breaks as biodosimetry for cell killing by carbon ions

We have studied the relationship between cell killing and the induction of residual chromatin breaks on various human cell lines and primary cultured cells obtained by biopsy from patients irradiated with either X-rays or heavy-ion beams to identify potential bio-marker of radiosensitivity for radiation-induced cell killing. The carbon-ion beams were accelerated with the Heavy Ion Medical Accelerator in Chiba (HIMAC). Six primary cultures obtained by biopsy from 6 patients with carcinoma of the cervix were irradiated with two different mono-LET beams (LET= 13 keV/μm, 76 keV/μm) and 200kV X rays. Residual chromatin breaks were measured by counting the number of non-rejoining chromatin fragments detected by the premature chromosome condensation (PCC) technique after a 24 hour postirradiation incubation period. The induction rate of residual chromatin breaks per cell per Gy was the highest for 76 keV/μm beams on all of the cells. Our results indicated that cell which was more sensitive to the cell killing was similarly more susceptible to induction of residual chromatin breaks. Furthermore there is a good correlation between these two end points in various cell lines and primary cultured cells. This suggests that the detection of residual chromatin breaks by the PCC technique may be useful as a predictive assay of tumor response to cancer radiotherapy.     

Genomic instability and tumorigenic induction in immortalized human bronchial epithelial cells by heavy ions

Carcinogenesis is postulated to be a progressive multistage process characterized by an increase in genomic instability and clonal selection with each mutational event endowing a selective growth advantage. Genomic instability as manifested by the amplification of specific gene fragments is common among tumor and transformed cells. In the present study, immortalized human bronchial (BEP2D) cells were irradiated with graded doses of either 1GeV/nucleon ⁵⁶Fe ions or 150 keV/μm alpha particles. Transformed cells developed through a series of successive steps before becoming tumorigenic in nude mice. Tumorigenic cells showed neither ras mutations nor deletion in the p16 tumor suppressor gene. In contrast, they harbored mutations in the p53 gene and over-expressed cyclin D1. Genomic instability among transformed cells at various stage of the carcinogenic process was examined based on frequencies of PALA resistance. Incidence of genomic instability was highest among established tumor cell lines relative to transformed, non-tumorigenic and control cell lines. Treatment of BEP2D cells with a 4 mM dose of the aminothiol WR-1065 significantly reduced their neoplastic transforming response to ⁵⁶Fe particles. This model provides an opportunity to study the cellular and molecular mechanisms involved in malignant transformation of human epithelial cells by heavy ions.     

Simulated microgravity inhibits the proliferation of K562 erythroleukemia cells but does not result in apoptosis

Astronauts and experimental animals in space develop the anemia of space flight, but the underlying mechanisms are still unclear. In this study, the impact of simulated microgravity on proliferation, cell death, cell cycle progress and cytoskeleton of erythroid progenitor-like K562 leukemia cells was observed. K562 cells were cultured in NASA Rotary Cell Culture System (RCCS) that was used to simulate microgravity (at 15 rpm). After culture for 24 h, 48 h, 72 h, and 96 h, the cell densities cultured in RCCS were only 55.5%, 54.3%, 67.2% and 66.4% of the flask-cultured control cells, respectively. The percentages of trypan blue-stained dead cells and the percentages of apoptotic cells demonstrated no difference between RCCS-cultured cells and flask-cultured cells at every time points (from 12 h to 96 h). Compared with flask-cultured cells, RCCS culture induced an accumulation of cell number at S phase concomitant with a decrease at G0/G1 and G2/M phases at 12 h. But 12 h later (from 24 h to 60 h), the distribution of cell cycle phases in RCCS-cultured cells became no difference compared to flask-cultured cells. Consistent with the changes of cell cycle distribution, the levels of intercellular cyclins in RCCS-cultured cells changed at 12 h, including a decrease in cyclin A, and the increasing in cyclin B, D1 and E, and then (from 24 h to 36 h) began to restore to control levels. After RCCS culture for 12–36 h, the microfilaments showed uneven and clustered distribution, and the microtubules were highly disorganized. These results indicated that RCCS-simulated microgravity could induce a transient inhibition of proliferation, but not result in apoptosis, which could involve in the development of space flight anemia. K562 cells could be a useful model to research the effects of microgravity on differentiation and proliferation of hematopoietic cells.