动物胚胎的玻璃化冻存

动物胚胎的玻璃化冻存是一项新的深低温冷冻方法,已成功地冻存了多种动物和人的胚胎,具有较高的实际应用和研究价值。本文结合本实验室的研究工作,论述玻璃化冻存方法在动物胚胎保存中的应用,不同保护剂和玻璃化溶液对胚胎存活率的影响,分析影响玻璃化冻存的主要因素包括冷冻保护剂与玻璃化溶液的选择、预平衡过程、冷却和复温速率控制、去玻璃化的避免、冻存后玻璃化溶液的洗脱等。     

动物组织细胞玻璃化冻存方法的研究进展

动物组织细胞的玻璃化冻存是一项新的深低温冷冻保存方法，已成功地冻存了多种动物细胞，具有较高的实际应用和研究价值。本文论述几种主要组织细胞玻璃化冻存方法的研究及其进展，如小鼠胰岛和卵母细胞，大鼠卵巢、牛胚泡、牛卵母细胞、猪胚胎、人胚胎等，不同冷冻保护剂，玻璃化溶液和冷冻程序对细胞存活率的影响。并提出了玻璃化冻存动物以及组织细胞的问题与展望。     

海冰离心脱盐理论与实验研究

海冰盐度主要来自海水结冰时被包裹在内部的卤水盐胞,海冰离心脱盐方法正是依据此,将海冰破碎,使卤水暴露出来,进而施加超重力,脱除卤水,淡化海冰。本文通过理论分析得出海冰离心脱盐后盐度与时间的理论关系式为S(t)=PK1/t,采用该式的计算结果与文献中的实验数据对比表明两者吻合较好。在理论分析基础上,通过实验确定了粒度、分离因数和加入海水比例对海冰离心脱盐的影响。实验结果表明,淡化盐度随着粒径呈现先减小后增大的趋势,并且当颗粒在6～9mm左右达到最小;随着加入海水比例和分离因数的增大,海冰离心脱盐效果变好。     

Thermomodulated cell culture∕harvest in polydimethylsiloxane microchannels with poly(N-isopropylacrylamide)-grafted surface

Cell culture and harvest are the most upstream operation for a completely integrated cell assay chip. In our previous work, thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) was successfully grafted onto polydimethylsiloxane (PDMS) surface via benzophenone-initiated photopolymerization. In the present work, the PNIPAAm-grafted-PDMS (PNIPAAm-g-PDMS) surface was explored for thermomodulated cell culture and noninvasive harvest in microfluidic channels. Using COS 7 fibroblast from African green monkey kidney as the model cells, the thermomodulated adhering and detaching behaviors of the cells on the PNIPAAm-g-PDMS surfaces were optimized with respect to PNIPAAm-grafting yields and gelatin modification. The viability of the cells cultured on and harvested from the PNIPAAm-g-PDMS surface with the thermomodulated noninvasive protocol was estimated against the traditional cell culture∕harvest method involving trypsin digestion. The configuration of the microchannel on the PNIPAAm-g-PDMS chip was evaluated for static cell culture. Using a pipette-shaped PNIPAAm-g-PDMS microchannel, long-term cell culture could be achieved at 37 °C with periodic change of the culture medium every 12 h. After moving the microchip from the incubator set at 37 °C to the room temperature, the proliferated cells could be spontaneously detached from the PNIPAAm-g-PDMS surface of the upstream chamber and transferred by a gentle fluid flow to the downstream chamber, wherein the transferred cells could be subcultured. The thermomodulated cell culture, harvest, and passage operations on the PNIPAAm-g-PDMS microfluidic channels were demonstrated.     

Mapping of hyperthermic tumor cell death in a microchannel under unidirectional heating

Hyperthermia can be used as an adjunctive method of chemotherapy, radiotherapy, and gene therapy to improve cancer treatment. In this study, we investigate the hyperthermic cell death of cervix cancer CaSki cells in a microchannel integrated with a directional heating scheme. Heat was applied from the inner end to the outer end of the channel and a temperature distribution from 60 °C to 30 °C was established. A three dimensional (3D) numerical model was conducted for the heat transfer simulation, based on which a simple fitting method was proposed to easily estimate the temperature distribution along the channel. Cell death along the channel was mapped 22 h after the heating treatment by dual fluorescent labeling and phase-contrast microscopy imaging. Upstream, where the temperature is higher than 42 °C, we observe necrotic death, late-stage and early stage apoptotic death in sequence along the channel. Downstream and in the middle of the channel, where the temperature is lower than 42 °C, significant cell detachment was noted. Vigorous detachment was observed even in the non-hyperthermic zone (temperature lower than 37 °C), which we believe is due to the direct effect of the hyperthermic zones (higher than 37 °C). The present work not only gives a vivid map of cell responses under a temperature gradient, but also reveals the potential interactions of the heated tumor cells and non-heated tumor cells, which are seldom investigated in conventional petri-dish experiments.     

黄河宁蒙河段水文-水温过程和河道形态变化对凌汛的影响

黄河宁蒙河段的水文和水温过程近几十年来发生了显著变化,迫切需要评估这一变化对河道形态、凌汛的影响及其可能的致灾效应。利用同水位河道过水断面形态参数分析方法,选取宁蒙河段稳定封冻区域（石嘴山-头道拐）的石嘴山、磴口、巴彦高勒、三湖河口和头道拐5个主要水文站,根据2008-2013年凌汛前实测断面资料,计算相应年份5个水文站同水位下的河道断面面积、平均水深和深泓点高程。结果表明,宁蒙河段近年来凌汛期的径流量和水温呈现逐渐增大的趋势,下泄流量过程有助于减少凌灾发生的几率;各水文站凌汛前的断面面积和平均水深等断面形态参数呈逐年增大的趋势,深泓点高程不断降低。河道过水断面形态的这种变化增大了凌汛期河道的过流能力,有助于缓解开河期可能出现的凌汛灾情。     

A numerical study on distributions during cryoprotectant loading caused by laminar flow in a microchannel

In this work, we conduct a computational study on the loading of cryoprotective agents into cells in preparation for cryopreservation. The advantages of microfluidics in cryopreserving cells include control of fluid flow parameters for reliable cryoprotectant loading and reproducible streamlined processing of samples. A 0.25 m long, three inlet T-junction microchannel serves as an idealized environment for this process. The flow field and concentration distribution are determined from a computational fluid dynamics study and cells are tracked as inert particles in a Lagrangian frame. These particles are not confined to streamlines but can migrate laterally due to the Segre-Sildeberg effect for particles in a shear flow. During this tracking, the local concentration field surrounding the cell is monitored. This data are used as input into the Kedem-Katchalsky equations to numerically study passive solute transport across the cell membrane. As a result of the laminar flow, each cell has a unique pathline in the flow field resulting in different residence times and a unique external concentration field along its path. However, in most previous studies, the effect of a spatially varying concentration field on the transport across the cell membrane is ignored. The dynamics of this process are investigated for a population of cells released from the inlet. Using dimensional analysis, we find a governing parameter α, which is the ratio of the time scale for membrane transport to the average residence time in the channel. For α <  = 0.224, cryoprotectant loading is completed to within 5% of the target concentration for all of the cells. However, for α > 0.224, we find the population of cells does not achieve complete loading and there is a distribution of intracellular cryoprotective agent concentration amongst the population. Further increasing α beyond a value of 2 leads to negligible cryoprotectant loading. These simulations on populations of cells may lead to improved microfluidic cryopreservation protocols where more consistent cryoprotective agent loading and freezing can be achieved, thus increasing cell survival.     

Effect of Joule heating on isoelectric focusing of proteins in a microchannel

Electric field-driven separation and purification techniques, such as isoelectric focusing (IEF) and isotachophoresis, generate heat in the system that can affect the performance of the separation process. In this study, a new mathematical model is presented for IEF that considers the temperature rise due to Joule heating. We used the model to study focusing phenomena and separation performance in a microchannel. A finite volume-based numerical technique is developed to study temperature-dependent IEF. Numerical simulation for narrow range IEF (6 < pH < 10) is performed in a straight microchannel for 100 ampholytes and two model proteins: staphylococcal nuclease and pancreatic ribonuclease. Separation results of the two proteins are obtained with and without considering the temperature rise due to Joule heating in the system for a nominal electric field of 100 V/cm. For the no Joule heating case, constant properties are used, while for the Joule heating case, temperature-dependent titration curves and thermo-physical properties are used. Our numerical results show that the temperature change due to Joule heating has a significant impact on the final focusing points of proteins, which can lower the separation performance considerably. In the absence of advection and any active cooling mechanism, the temperature increase is the highest at the mid-section of a microchannel. We also found that the maximum temperature in the system is a strong function of the ΔpK? value of the carrier ampholytes. Simulation results are also obtained for different values of applied electric fields in order to find the optimum working range considering the simulation time and buffer temperature. Moreover, the model is extended to study IEF in a straight microchip where pH is formed by supplying H⁺ and OH⁻, and the thermal analysis shows that the heat generation is negligible in ion supplied IEF.     

Acoustofluidic phase microscopy in a tilted segmentation-free configuration

A low-cost device for registration-free quantitative phase microscopy (QPM) based on the transport of intensity equation of cells in continuous flow is presented. The method uses acoustic focusing to align cells into a single plane where all cells move at a constant speed. The acoustic focusing plane is tilted with respect to the microscope’s focal plane in order to obtain cell images at multiple focal positions. As the cells are displaced at constant speed, phase maps can be generated without the need to segment and register individual objects. The proposed inclined geometry allows for the acquisition of a vertical stack without the need for any moving part, and it enables a cost-effective and robust implementation of QPM. The suitability of the solution for biological imaging is tested on blood samples, demonstrating the ability to recover the phase map of single red blood cells flowing through the microchip.     

镁合金薄板差温拉深成形模具温度控制系统设计

凸模、凹模、压边圈各自温度及其分布的均匀性是镁合金薄板差温拉深成形加工过程中需控制的关键技术之一,将直接影响镁合金板材差温拉深效果、模具使用寿命.设计了电阻环形线圈加热凸模和压边圈、室温冷却水对凸模进行冷却、PID温度控制器控温的镁合金差温拉深模具温度控制系统.使用表明,该系统反馈及时、温控精度高、模具温度分布均匀,提高了镁合金薄板加热质量.     

蚯蚓资源在海冰水改良渤海湾滨海盐渍土壤中的应用前景

海冰在其成冰过程中盐分大量析出而盐度较低,可能成为干旱季节的一种灌溉水资源。海冰水改良滨海盐渍土是在渤海湾地区利用海冰融解的低盐水灌溉和淋洗高盐土壤的一种措施,该措施应用的同时也产生土壤熟化时间慢、土壤结构恶化、土壤养分的流失和土壤生物种群数量减少等问题。本文在收集和分析大量资料基础上,论述了蚯蚓与盐渍土壤的相互关系:盐碱地土壤中蚯蚓种类和盐渍土壤对蚯蚓的影响因素。根据在盐渍土壤中,蚯蚓活动能够增加有机物的分解和养分释放,同时使盐碱地土壤的团聚体结构、渗透率、生物数量和活性得以恢复和改善,进一步阐述了在海冰水改良滨海盐渍土壤中引入蚯蚓的可行性,并探讨引入蚯蚓改良滨海盐渍土壤的研究方法和存在的问题以及今后的研究方向。     

自然冷冻法处理生活污水的研究初探

本文通过人工模拟气候下的冷冻实验,以生活污水为研究对象,分析冷冻法对水中有机物和NH3-N的去除效果。实验结果表明:冷冻法能有效的去除水中的污染物,对COD去除效率达到80%左右,对NH3-N去除效率达到90%左右。本方法所需设备简单、操作简便、高效低耗,特别适合我国北方冬季长、气温低的气候条件。     

Effect of surface acoustic waves on the viability, proliferation and differentiation of primary osteoblast-like cells

Surface acoustic waves (SAWs) have been used as a rapid and efficient technique for driving microparticles into a three-dimensional scaffold matrix, raising the possibility that SAW may be effective in seeding live cells into scaffolds, that is, if the cells were able to survive the infusion process. Primary osteoblast-like cells were used to specifically address this issue: To investigate the effects of SAW on the cells’ viability, proliferation, and differentiation. Fluorescence-labeled osteoblast-like cells were seeded into polycaprolactone scaffolds using the SAW method with a static method as a control. The cell distribution in the scaffold was assessed through image analysis. The cells were far more uniformly driven into the scaffold with the SAW method compared to the control, and the seeding process with SAW was also significantly faster: Cells were delivered into the scaffold in seconds compared to the hour-long process of static seeding. Over 80% of the osteoblast-like cells were found to be viable after being treated with SAW at 20 MHz for 10–30 s with an applied power of 380 mW over a wide range of cell suspension volumes (10–100 μℓ) and cell densities (1000–8000 cells∕μℓ). After determining the optimal cell seeding parameters, we further found that the treated cells offered the same functionality as untreated cells. Taken together, these results show that the SAW method has significant potential as a practical scaffold cell seeding method for tissue and orthopedic engineering.     

A microfluidic-based neurotoxin concentration gradient for the generation of an in vitro model of Parkinson's disease

In this study, we developed a miniaturized microfluidic-based high-throughput cell toxicity assay to create an in vitro model of Parkinson's disease (PD). In particular, we generated concentration gradients of 6-hydroxydopamine (6-OHDA) to trigger a process of neuronal apoptosis in pheochromocytoma PC12 neuronal cell line. PC12 cells were cultured in a microfluidic channel, and a concentration gradient of 6-OHDA was generated in the channel by using a back and forth movement of the fluid flow. Cellular apoptosis was then analyzed along the channel. The results indicate that at low concentrations of 6-OHDA along the gradient (i.e., approximately less than 260 μM), the neuronal death in the channel was mainly induced by apoptosis, while at higher concentrations, 6-OHDA induced neuronal death mainly through necrosis. Thus, this concentration appears to be useful for creating an in vitro model of PD by inducing the highest level of apoptosis in PC12 cells. As microfluidic systems are advantageous in a range of properties such as throughput and lower use of reagents, they may provide a useful approach for generating in vitro models of disease for drug discovery applications.     

黄渤海海冰分布特征及其影响因子

海冰是中国黄渤海冬季主要海洋灾害之一。为进一步认识黄渤海海冰分布特征和日变化机理,提高海冰短期预报水平,本文利用美国国家冰雪数据中心（NSIDC）MASIE海冰范围产品和欧洲中期天气预报中心（ECMWF）ERA-interim再分析资料,统计了2018年冬季黄渤海海冰时空分布特征,并通过相关性分析,研究了日海冰面积与海洋气象因子的关系。结果表明：2018年冬季黄渤海海冰集中在辽东湾和朝鲜湾,海冰覆盖率随离岸距离的增加而降低;海冰变化经历了生成、维持和融化3个阶段,其中生成阶段明显长于融化阶段。39°N以北黄渤海日均海表温度（SST）、海面2 m气温（T_2m）、海面2 m气温<-2℃积温（T_c）和海冰面积呈显著负相关关系,Pearson相关系数分别为-0.61（P<0.01）、-0.52（P<0.01）和-0.50（P<0.01）。因此,39°N以北黄渤海海气界面的热力环境是影响黄渤海海冰面积的重要因素。另外研究发现在2018年黄渤海海冰生成发展阶段,海冰面积共出现两次异常减少,这两次日海冰面积异常减少主要是由海区前一日升温和偏南风造成的。     

综合平衡法在浸渍-升华工序工艺参数分析中的应用

为了提高干冰膨胀烟丝质量,针对浸渍-升华工序对膨胀后烟丝质量的影响,文章采用正交试验设计与综合平衡法对升华温度、蒸汽流量、塔内风速、浸渍时间4个工艺参数进行了分析,结果表明:在试验范围内,升华温度320℃,蒸汽流量800-1000 kg/h、塔内风速34 m/s,浸渍时间30秒,为最佳工艺参数,按此参数生产的膨胀烟丝综合质量最优。     

红外热成像技术在研究不同果蔬真空预冷特性中的应用

选取表面结构存在较大差异的三种蔬菜:番茄(Lycopersicum esculentum)、双孢菇(Agaricus bisporus)、青菜(Brassica campestris L.ssp.chinensis(L.)Makino.var.communis Tsen et Lee)作为研究对象,利用红外热成像技术在终压为300Pa的真空预冷中.研究它们表面温度的变化情况.结果表明:双孢菇表面降温效果最好,青菜次之,番茄最差;而且双孢菇在预冷过程中,表面会有过冷现象发生:青菜的叶部与茎部相比,叶部的降温速率和复压阶段的升温速率都较快;此外,在青菜表面的轻微损伤处出现了局部低温.     

A new three-dimensional heat flux-temperature integral relationship for half-space transient diffusion

This paper derives a new integral relationship between heat flux and temperature in a transient, three-dimensional heat conducting Cartesian half space (x>0, y∈(−∞,∞), z∈(−∞,∞)). A unified mathematical treatment has been developed based on operational and transform methods; and singular integral equation regularization. Regularization is accomplished based on a series of observations involving the diffusive nature of the operator. This newly developed relationship provides the local heat flux perpendicular to the front surface at any location within the half space. This expression suggests that an embedded plane of temperature sensors parallel to the surface can be used to acquire the local, in-depth heat flux in the x-direction. The relationship does not require a priori knowledge of the surface boundary condition which has analytically been removed in the process. The ill-posed nature of diffusion is highlighted owing to the appearance of the heating/cooling rate (°C/s) in the integrand of the new relationship. Integral relationships of this type are highly useful for experimental investigations since the in-depth heat flux can be extracted from well-established temperature transducers.     

Characterization of low viscosity polymer solutions for microchip electrophoresis of non-denatured proteins on plastic chips

In this paper, we study characteristics of polymers (methylcellulose, hypromellose ((hydroxypropyl)methyl cellulose), poly(vinylpyrrolidone), and poly(vinyl alcohol)) with different chemical structures for microchip electrophoresis of non-denatured protein samples in a plastic microchip made of poly(methyl methacrylate) (PMMA). Coating efficiency of these polymers for controlling protein adsorption onto the channel surface of the plastic microchip, wettability of the PMMA surface, and electroosmotic flow in the PMMA microchannels in the presence of these polymers were compared. Also relative electrophoretic mobility of protein samples in solutions of these polymers was studied. We showed that when using low polymer concentrations (lower than the polymer entanglement point) where the sieving effect is substantially negligible, the interaction of the samples with the polymer affected the electrophoretic mobility of the samples. This effect can be used for achieving better resolution in microchip electrophoresis of protein samples.     

篦子三尖杉的胚胎学研究及其系统位置的探讨

篦子三尖杉Cephalotaxus oliveri的成熟花粉包含两个细胞，花粉管在精原细胞产生后 经历了一年的休眠期; 精原细胞分裂产生两个体积不等的精子; 雌配子体游离核经12次分裂后形成细胞壁; 窄长的颈卵器单个顶生; 卵细胞中含有大量类核仁状体; 受精时，雌雄核在分裂中期完全融合; 原胚游离核在16核时产生细胞壁; 原胚具冠细胞; 无裂生多胚现象; 成熟胚 二枚子叶。总之，蓖子三尖杉与本属其它种相比，存在明显的差异。如精原细胞分裂前细胞中心细 胞质浓缩成星状放射区域; 苗端早在胚成熟前分化形成明显的突起，以及成熟胚的子叶极为发达等等。因此，作者赞同将蓖子三尖杉在本属中另立一组的观点。