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在过去的半个世纪，细胞生物学和分子生物学是通过细胞培养和胞外分析来研究基因和蛋白等细胞成分的。而绿色荧光蛋白作为报告基因出现后，人类将研究的基因或蛋白与绿色荧光蛋白连接，通过观测荧光变化直接动态观察细胞生物学和分子生物学的变化。荧光蛋白转染至肿瘤细胞系中，如肺癌、膀胱癌，并不改变肿瘤细胞侵袭和转移的特性。

Macromolecular assemblies that regulate chromatin structure using the energy of ATP hydrolysis have critical roles in development, cancer, and stem cell biology. The ATPases of this family are encoded by 27 human genes and are usually associated with several other proteins that are stable, non-exchangeable subunits. One fundamental mechanism used by these complexes is thought to be the movement or exchange of nucleosomes to regulate transcrip- tion. However, recent genetic studies indicate that chromatin remodelers may also be involved in regulating other aspects of chromatin structure during many cellular processes. The SWI/SNF family in particular appears to have undergone a substantial change in subunit composition and mechanism coincident with the evolutionary advent of multicellularity and the appearance of linking histones. The differential usage of this greater diversity of mammalian BAF subunits is essential for the development of specific cell fates, including the progression from pluripotency to multipotency to committed neurons. Recent human genetic screens have revealed that BRG1, ARID1A, BAF155, and hSNF5 are frequently mutated in tumors, indicating that BAF complexes also play a critical role in the initiation or progression of cancer. The mechanistic bases underlying the genetic requirements for BAF and other chromatin remodelers in development and cancer are relatively unexplored and will be a focus of this review.

Dear Editor, Visualization of chromosome shapes and dynamics in a live cell is highly desirable and necessary in many areas of cell biology. For example, the copy number of a particular chromosome in cancer cells is often abnormal （e.g., more than two）, and therefore probing chromosome copy numbers can aid cancer diagnosis.

Eukaryotic organisms have quality-control mechanisms that allow misfolded or unassembled proteins to be re- tained in the endoplasmic reticulum （ER） and subsequently degraded by ER-associated degradation （ERAD）. The ERAD pathway is well studied in yeast and mammals; however, the biological functions-of plant ERAD have not been reported. Through molecular and cellular biological approaches, we found that ERAD is necessary for plants to overcome salt stress. Upon salt treatment ubiquitinated proteins increased in plant cells, especially unfolded proteins that quickly accumulated in the ER and subsequently induced ER stress responses. Defect in HRD3A of the HRD1/ HRD3 complex of the ERAD pathway resulted in alteration of the unfolded protein response （UPR）, increased plant sensitivity to salt, and retention of ERAD substrates in plant ceils. Furthermore, we demonstrated that Ca2＋ release from the ER is involved in the elevation of UPR and reactive oxygen species （ROS） participates the ERAD-related plant salt response pathway.

The Hippo pathway is a signal transduction pathway that regulates organ growth, stem cell biology, regeneration and cancer. Three recent proteomic studies with Hippo path- way components uncovered extensive networks of interacting proteins revealing novel connections to cellcell junctions, regulation by vesicle trafficking, and phosphorylation- dependent remodeling of the interactome, and provide a rich landscape of novel interactors ripe for mechanistic studies.

The cysteine-rich secretory proteins （CRISPs） are a subgroup of the CRISP, antigen 5 and Pr-1 （CAP） protein superfamily, and are found only in vertebrates. They show a strong expression bias to the mammalian male reproductive tract and the venom of poisonous reptiles. Within the male reproductive tract CRISPs have been implicated in many aspects of male germ cell biology spanning haploid germ cell development, epididymal maturation, capacitation, motility and the actual processes of fertilization. At a structural level, CRISPs are composed of two domains, a CAP domain, which has been implicated in cell-cell adhesion, and a CRISP domain, which has been shown to regulate several classes of ion channels across multiple species. Herein, we will review the current literature on the role of CRISPs in male fertility, and by inference to related non-mammalian protein, infer potential biochemical functions.

Stem cells, including both pluripotent stem cells and multipotent somatic stem cells, hold great potential for interrogating the mechanisms of tissue development, homeostasis and pathology, and for treating numerous devastating diseases. Establishment of in vitro platforms to faithfully maintain and precisely manipulate stem cell fates is essential to understand the basic mechanisms of stem cell biology, and to translate stem cells into regenerative medicine. Chemical approaches have recently provided a number of small molecules that can be used to control cell selfrenewal, lineage differentiation, reprogramming and regeneration. These chemical modulators have been proven to be versatile tools for probing stem cell biology and manipulating cell fates toward desired outcomes. Ultimately, this strategy is promising to be a new frontier for drug development aimed at endogenous stem cell modulation.

GCaMP is one of the most widely used calcium indicators in neuronal imaging and calcium cell biology. The newly developed GCaMP6 shows superior brightness and ultrasensitivity to calcium concentration change. In this study, we determined crystal structures of CaZ＋-bound GCaMP6 monomer and dimer and presented detailed structural analyses in comparison with its par- ent version GCaMP5G. Our analyses reveal the structural basis for the outperformance of this newly developed Ca2＋ indicator. Three substitution mutations and the resulting changes of local structure and interaction explain the ultrasensitivity and in- creased fluorescence intensity common to all three versions of GCaMP6. Each particular substitution in the three GCaMP6 is also structurally consistent with their differential sensitivity and intensity, maximizing the potential of using GCaMP6 in solving diverse problems in neuronal research and calcium signaling. Our studies shall also be beneficial to further structure-guided optimization of GCaMP and facilitate the design of novel calcium indicators.

The foundations of proteomics are to study gene products and their regulatory roles within cells. Paradoxically, the only evidence that sperm cells make new proteins is through mitochondrial protein synthesis. Yet despite this, spermatozoa are the perfect candidates for mass spectrometry and hence, proteomic analysis. These enterprising cells use a plethora of post-translational modifications in order to gain functionality following their production within the testis. By using a combination of two-dimensional polyacrylamide gel electrophoresis （2D-PAGE）, and more recently liquid chromatography-mass spectrometry （LC-MS）/MS, recent advances in sperm cell biology, through the use of proteomics, is making unparalleled progress. The protein inventory lists being generated have shed light on transmembrane proteins, kinases and chaperones never previously recognized. In addition, the ability to isolate either phosphopeptides or glycopeptides and quantify the differences between cells of two different populations make proteomic analysis of spermatozoa a real chance to finally answer some age old questions.

Study of cell-in-cell phenomena began with the desire to understand the complex phenotype, which stemmed from Lewis＇ observation ofnon-phagocytotic process of cell-eating-cell in 1925. Humble et al. coined the term ＂emperipolesis＂ （from the Greek, wandering round about within） in 1950s, to define the heterogeneous cell-in-cell phenomena when they studied bio- logical interaction of lymphocytes with other cells . Emperipolesis has since been found to be commonly enacted by lymphocytes in physiological and pathophysiological settings . Since the term ＂emperipolesis＂ was annotated, a challenge has been set to understand how a cell inhabits in the other, what are their respective fates, and the biological relevance of co-habitation.