细胞行为组学:一种从系统论的角度进行中医科学化的研究策略

中医的现代化以及标准化需要中西医结合的手段来实现。然而,中医研究强调整体观,目前的生物技术却无法有效的整合由分子生物学主导的基因组学、代谢组学以及转录组学等各种分子组学所产生的信息来完整的描述组织生理。与此同时,在单细胞层面,细胞已经完成对不同时空背景的复杂蛋白网络信息的整合,并将其表现在各种细胞行为上。在此,作者提出以细胞行为组学为主的策略并简短的介绍如何运用这种策略来进行针刺启动以及中药方剂的研究。

茯苓多糖衍生碳点的细胞行为学研究

碳点的生物学行为,特别是关于细胞摄取和细胞内分布机制研究,是其生物医学应用的基础。本研究以茯苓多糖为原料,采用水热法合成了蓝色荧光碳量子点,探索碳点进入细胞的具体生物学行为,以评估其生物学活性。利用透射电子显微镜、紫外-可见吸收光谱、荧光光谱、傅里叶变换红外光谱、X射线衍射和X射线光电子能谱对其进行了表征;以免疫细胞-RAW264.7细胞(小鼠单核巨噬细胞)和癌细胞-4T1细胞(小鼠乳腺癌细胞)这两种不同的细胞系为研究对象,研究茯苓多糖碳点在细胞内的摄取动力学过程及其摄取途径、分布和外排等过程。研究结果显示,茯苓多糖衍生碳点的粒径分布范围为2~10 nm,平均粒径为6.85 nm,主要由C、O和N元素组成,具有-OH、C=O、C-N、C=C等丰富的表面官能团,荧光量子产率为4.72%。碳点进入细胞可呈现出一定的浓度和时间依赖性,不同的细胞系,摄取内吞途径不一。在RAW264.7细胞内主要以巨噬细胞特有的吞噬作用摄取进入细胞内,少部分通过小窝蛋白介导的内吞作用,而对于4T1细胞主要是通过网格蛋白介导的内吞作用和巨胞饮过程。综上,合成的中药衍生碳点具有良好的荧光特性、低细胞毒性和优异的生物相容性,可用于细胞成像应用研究。

针灸作用机制研究的困惑与出路--针灸调节肠易激综合征肠功能机制研究的系统科学思考

以还原论为主体的研究方法在针刺作用机制探索过程中发挥了巨大作用,但该方法也存在明显的局限性,以针灸调节肠易激综合征(IBS)肠功能机制研究为例,对此进行了反思。认为针灸治疗IBS机制的探索涉及范围广泛、层次多样,但获得的认识多是碎片化的,缺乏对众多环节间相互作用和相互协调的了解。为弥补还原论方法的不足,从系统科学角度探索针灸的作用机制是一种必然的趋势,从系统生物学的角度,特别是从细胞行为组学角度对针灸调节机制的研究思路及其可行性进行探讨,提出了一是基于“中枢神经系统-自主神经系统-胃肠道神经-Cajal间质细胞-平滑肌细胞(CNS-ANS-ENS-ICC-SMC)网络”细胞行为组学的研究,二是基于肠道类器官多种细胞行为组学的研究,以期将针灸调节规律及其机制的认识推进到更深的层次。

中西医结合研究的思维及方法学亟待创新——运用细胞行为组学进行中医药研究的探讨

组学技术在近年来中西医结合的发展中发挥了重要作用。由于生理或病理状态是由生命体最基本的单位:细胞的行为表现聚合而形成的,所以凭借各种组学所产生的分子信息必须在细胞层面加以整合,并延展到人体整体层面才能体现中医学理论的精髓。细胞行为是细胞内蛋白网络中所有蛋白活性受到外界调控后在细胞层面的总体表现。精细量化细胞的各种行为可以解析不同蛋白通路在细胞行为上的占比,并将各种分子组学信息和细胞行为紧密连结。笔者首次倡导基于细胞行为组学来研究中医药生物学基础的策略,旨在探寻适合中医学理论的研究方法,以全面体现中医学理论内涵及其实践在现代科学中的意义。

Expression of a novel apoptosis inhibitor-survivin in colorectal carcinoma

AIM： To investigate the role of survivin expression in the pathogenesis of colorectal carcinoma. METHODS： Immunohistochemistry S-P method and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNE） were used to detect the expression of survivin and apoptotic cell in situ in colorectal cancerous tissues, para-cancerous tissues and normal tissues of 48 cases of colorectal carcinoma. RESULTS： The survivin positive unit （PU） was higher in cancerous tissues （38.76±5.14） than in para-cancerous （25.17±7.26） or normal tissues （0.57±0.03） （P〈0.05）. The apoptosis index （AI） of para-cancerous tissues was （7.51±2.63%） higher than cancerous tissues （4.65±1.76%）. The expression of survivin was associated with pathological grade, lymph node metastasis and Dukes stage of colorectal carcinoma. CONCLUSION： Survivin expression may play an important role in carcinogenesis of colorectal carcinoma and may be associated with malignant biological behaviors of colorectal carcinoma.

ZNF217 expression correlates with the biological behavior of human ovarian cancer cells

The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.

Effects of lobaplatin and oxaliplatin on biological behavior of colorectal carcinoma cell line

Objective： The aim of the study was to observe the effects of Iobaplatin and oxaliplatin on biological behavior of colorectal carcinoma cell line. Methods： The human colorectal carcinoma cell line LoVo and HR-8348 were used in the present study, and different concentrations of Iobaplatin and oxaliplatin served as inhibitors. The invasion and metastasis potential were evaluated with Transwell chamber, and cell inhibition ratio was measured with MTT assay. Results： Lobaplatin and oxaliplatin could significantly decrease the abilities of invasion and metastasis of LoVo and HR-8348 as compared with the control group （P 〈 0.01）, but no difference between Iobaplatin and oxaliplatin groups was observed （P 〉 0.05）. No difference was found in the cell inhibition ratio between the Iobaplatin and oxaliplatin groups （P 〉 0.05）. Lobaplatin could inhibit colorectal carcinoma cells that oxaliplatin couldn＇t do. Conclusion： Lobaplatin is a new antitumor platinum drug and can inhibit colorectal carcinoma cells. Moreover, Iobaplatin could also inhibit colorectal carcinoma cells which are resistant to oxaliplatin.

Atypical glomus tumor of the body of stomach: a case report and review of literature

Objective: The aim of this study was to analyze the clinical pathological features of the atypical glomus tumor of the stomach, and explore its biological behavior and clinical significance. Methods: a 43-year-old female was admitted due to passing black stool for 5 months. The melena of the patient occurred after eating the crabs. The endoscopic ultrasonography (EUS) was performed, showing a hypoecho nodule found at the body of the stomach, suggesting the diagnosis of gastric stromal tumor. After onset of disease, no specific clinical symptoms were observed except for the slight feeling discomfort in upper abdominal region. The gastroscopy was performed under the general anesthesia: there was a spheroidal nodule measuring 2.5 cm x 3.0 cm at the anterior wall of the body of stomach near the side of lesser curvature with the focal erosion of gastric mucosa. The DualknifeTM and the snare were used to excise the tumor in full thickness. Grossly, the tumor nodule measured 2.5 cm in diameter. The cut surface was solid, showing moderate consistency and grey red in colour. Microscopically, this tumor presented a well circumscribed nodule. It composed of the rounded neoplastic cells which were mainly arranged in solid pattern. The tumor cells had well-defined cell membranes, centrally placed, round nuclei and amphophilic cytoplasm. Immunohistochemical staining showed the positive expression of smooth muscle actin (SMA), caldesmon and vimentin; meanwhile detecting the negative expression of desmin, pan-cytokeratin, CD34, CD117 and S-100 in tumor cells. The Ki-67 labelling index of tumor was 8%. Results: Combined with the histopathological features, immunophenotype of the tumor, deep location in abdominal cavity and with greater diameter larger than 2 cm, and the diagnosis of atypical glomus tumor of the body of the stomach in this case was established. Conclusion: The atypical glomus tumor of the body of the stomach is a rare mesenchymal tumor. The following differential diagnosis should be included as following: gastrointestinal stromal tumor (GIST), paraganglioma, myopericytoma and the angioleiomyoma. The immunohistochemistry exerts an essential role during the diagnosis of this tumor. The deeper location and larger size implicate its undetermined biological behavior.

Influence on Biological Behavior of Colon Cancer Stem Cells after RNA Interfering CD133

OBJECTIVE To observe the effects expression and activation on biological cancer stem cells. of blocking CD133 gene characteristic of the colon METHODS CD133＋ colon cancer stem cells （CCSCs） were separated from EpCAMhigh CD44＋ CCSCs through fluorescenceactivated cell sorting （FACS）. The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133＋ CCSCs were observed after the CD133＋ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44＋ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot. RESULTS CD133＋ CCSCs were isolated from EpCAMhigh CD44＋ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133＋ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn＇t generate spherical cells. Results of MTT assay showed that the CD133＋ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133＋ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly （P 〈 0.01）. CD133- cells were obtained from the EpCAMhigh CD44＋ CCSCs using FACS, in which the expression of CD133 protein was positive. CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.

Cooperative Effects of Noise and Coupling on Stochastic Dynamics of a Membrane-Bulk Coupling Model

Based on a membrane-bulk coupling cell model proposed by Gomez-Marin et al. [ Phys. Rev. Lett. 98 （2007） 168303], the cooperative effects of noise and coupling on the stochastic dynamical behavior are investigated. For parameters in a certain region, the oscillation can be induced by the cooperative effect of noise and coupling. Whether considering the coupling or not, corresponding coherence resonance phenomena are observed. Furthermore, the effects of two coupling parameters, cell size L and coupling intensity k, on the noise-induced oscillation of membranes are studied. Contrary effects of noise are found in and out of the deterministic oscillatory regions.

Sequence analysis and functional study of the Han Nationality glial cell line-derived neurotrophic factor transcript

Objective: To study the sequence and function of the glial cell line-derived neurotrophic factor (GDNF) transcript in subjects of Han nationality. Methods: The Han nationality GDNF transcript was amplified by RT-PCR and expressed by baculovirus expression system. Biological activity of the expressed product was measured by the primary culture of midbrain dopaminergic neurons. Results: There only existed the shorter GDNF transcript of 555 bp in the Han nationality. The secretory expression product of the shorter transcript in insect cells promoted the survival and differentiation of dopaminergic neurons. Conclusion: It is found that there is a 78 bp deletion in the Han nationality GDNF transcript compared with the reported 633 bp GDNF transcript. The 78 bp deletion does not affect the secretory expression and biological activity of GDNF mature protein.

Sacro-anterior haemangiopericytoma: a case report

Haemangiopericytoma(HPC) is a rare vascular tumor with borderline malignancy, considerable histological variability, and unpredictable clinical and biological behavior. HPC can present a diagnostic challenge because of its indeterminate clinical, radiological, and pathological features. HPC generally presents in adulthood and is equally frequent in both sexes. HPC can arise in any site in the body as a slowly growing and painless mass. The precise cell type origin of HPC is uncertain. One third of HPCs occur in the head and neck areas. Exceptional cases of hemangioblastoma arising outside the head and neck areas have been reported, but little is known about their clinicopathologic and immunohistochemical features. This study reports on a case of a large sacro-anterior HPC in a 65-year-old male.

A review for dynamics of collective behaviors of network of neurons

The nervous system is composed of a large number of neurons, and the electrical activities of neurons can present multiple modes during the signal transmission between neurons by changing intrinsic bifurcation parameters or under appropriate external forcing. In this review, the dynamics for neuron, neuronal network is introduced, for example, the mode transition in electrical activity, functional role of autapse connection, bifurcation verification in biological experiments, interaction between neuron and astrocyte, noise effect, coherence resonance, pattern formation and selection in network of neurons. Finally, some open problems in this field such as electromagnetic radiation on electrical activities of neuron, energy consumption in neurons are presented.

Mechanical behavior of pathological and normal red blood cells in microvascular flow based on modified level-set method

The research of the motion and deformation of the RBCs is important to reveal the mechanism of blood diseases. A numerical method has been developed with level set formulation for elastic membrane immersed in incompressible fluid. The numerical model satisfies mass and energy conservation without the leaking problems in classical Immersed Boundary Method(IBM), at the same time, computing grid we used can be much smaller than the general literatures. The motion and deformation of a red blood cell(including pathological & normal status) in microvascular flow are simulated. It is found that the Reynolds number and membrane's stiffness play an important role in the transmutation and oscillation of the elastic membrane. The normal biconcave shape of the RBC is propitious to create high deformation than other pathological shapes. With reduced viscosity of the interior fluid both the velocity of the blood and the deformability of the cell reduced. With increased viscosity of the plasma both the velocity of the blood and the deformability of the cell reduced. The tank treading of the RBC membrane is observed at low enough viscosity contrast in shear flow. The tank tread fixed inclination angle of the cell depends on the shear ratio and viscosity contrast, which can be compared with the experimental observation well.

Mechanical behavior of the erythrocyte in microvessel stenosis

The passage of red blood cells （RBCs） through capillaries is essential for human blood microcirculation. This study used a moving mesh technology that incorporated leader-follower pairs to simulate the fluid-structure and structure-structure interac- tions between the RBC and a microvessel stenosis. The numerical model consisted of plasma, cytoplasm, the erythrocyte membrane, and the microvessel stenosis. Computational results showed that the rheology of the RBC is affected by the Reynolds number of the plasma flow as well as the surface-to-volume ratio of the erythroeyte. At a constant inlet flow rate, an increased plasma viscosity will improve the transit of the RBC through the microvessel stenosis. For the above reasons, we consider that the decreased hemorheology in microvessels in a pathological state may primarily be attributed to an increase in the number of white blood cells. This leads to the aggregation of RBCs and a change in the blood flow structure. The present fundamental study of hemorheology aimed at providing theoretical guidelines for clinical hemorheology.

Up-regulation of mitochondrial antioxidation signals in ovarian cancer cells with aggressive biologic behavior

Objective： Recently, a high frequency of mutations in mitochondrial DNA （mtDNA） has been detected in ovarian cancer. To explore the alterations of proteins in mitochondria in ovarian cancer, a pair of human ovarian carcinoma cell lines （SKOV3/SKOV3.ip1） with different metastatic potentials was examined. Methods： Cancer cells SKOV3.ipl were derived from the ascitic tumor cells of nude mice bearing a tumor of ovarian cancer cells SKOV3. SKOV3.ipl exhibited a higher degree of migration potential than its paired cell line SKOV3. The proteins in the mi- tochondria of these two cells were isolated and separated by 2-D gel electrophoresis. The differently expressed proteins were extracted and identified using matrix assisted laser desorption ionisation/time-of-flight/time-of-flight （MALDITOF/TOF）, and finally a selected protein candidate was further investigated by immunohistochemistry （IHC） method in nude mice bearing tumor tissues of these two cells. Results： A total of 35 spots with different expressions were identified between the two cells using 2D-polyacrylamide gel electrophoresis （PAGE） approach. Among them, 17 spots were detected only in either SKOV3 or SKOV3.ipl cells. Eighteen spots expressed different levels, with as much as a three-fold difference between the two cells. Twenty spots were analyzed using MALDI-TOF/TOF, and 11 of them were identified successfully; four were known to be located in mitochondria, including superoxide dismutase 2 （SOD2）, fumarate hydratase （FH）, mitochondrial ribosomal protein L38 （MRPL38）, and mRNA turnover 4 homolog （MRTO4）. An increased staining of SOD2 was observed in SKOV3.ipl over that of SKOV3 in IHC analysis. Conclusions： Our results indicate that the enhanced antioxidation and metabolic potentials of ovarian cancer cells might contribute to their aggressive and metastatic behaviors. The underlying mechanism warrants further study.

GLOBAL DYNAMICS OF AN IN-HOST HIV-HNFECTION MODEL WITH THE LONG-LIVED INFECTED CELLS AND FOUR INTRACELLULAR DELAYS

In this paper, we investigate global dynamics for an in-host HIV-1 infection model with the long-lived infected cells and four intracellular delays. Our model admits two possible equilibria, an uninfected equilibrium and infected equilibrium depending on the basic reproduction number. We derive that the global dynamics are completely determined by the values of the basic reproduction number： if the basic reproduction number is less than one, the uninfected equilibrium is globally asymptotically stable, and the virus is cleared; if the basic reproduction number is larger than one, then the infection persists, and the infected equilibrium is globally asymptotically stable.