Abstract:Experimental animals and human disease research are a symbiotic relationship in life science field. Due to the proximity with humans in the aspects of genetic evolution, physiology, biochemistry and pathology, macaque monkeys become indispensable experimental animals in biomedicine research. Genetically modified macaques (also known as genetic engineering macaques) could be important experimental materials and advanced animal models for understanding molecular mechanisms of human physiology and pathology under observation of expression and being regulation of the targeted gene modified in the whole life of animals. Currently, combination of high-efficient assisted reproductive technologies (ARTs) in macaques with precise gene editing technologies is the only way of developing genetically modified monkeys. Now in China, ARTs in Chinese rhesus macaques included semen collection and cryopreservation, superovulation, in vitro fertilization and embryo culture, embryo transfer and pregnancy management, thus "test-tube monkey", and all have been efficiently completed. The establishment and development of the current gene editing technology, ZFN (zinc-finger nucleases), TALEN (transcription activator-like effector nucleases) and CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/Cas9), create great chances for production of genetically modified macaques. Scientists in China have successfully developed transgenic macaques with overexpression of green fluorescence protein gene and gene mutagenesis in macaques. With the breakthrough and development of various technologies, combined with abundant experimental macaque resources, avoiding various reproductive physiological obstructions of macaques, China is expected to become an appropriate international base in application of genetic engineering macaques.

Abstract:In recent years, the cytokine hypothesis of depression has received considerable research attention, providing insights on the mechanism study and the therapeutic treatment of depressive disorders. Two classes of cytokines, pro-inflammatory cytokines and anti-inflammatory cytokines, are proven to be associated with depression. In this review, the focus is on the research of anti-inflammatory cytokines, which are believed to have a key role in alleviating depression-related symptoms. Anti-inflammatory cytokines, such as interleukin-10, interleukin-1 receptor antagonist, interleukin-4, interleukin-13, transforming growth factor-β, and adiponectin, show decreased concentrations during depressive episodes. Anti-inflammatory cytokines participate in the regulation mechanism of depression through antagonizing pro-inflammatory cytokines and interacting with MAPK signaling, neurotransmitters, and glucocorticoids. Various kinds of antidepressants can lead to an increased secretion of anti-inflammatory cytokines, which might be the action mechanism of antidepressants. Overall, accumulating evidences from the literature demonstrate positive prospects of using anti-inflammatory cytokines as a therapeutic alternative to treat depression.

Abstract:For any protein, its 3-D structure and interaction with other proteins are vital information for the understanding of its function. Traditionally, NMR and X-ray crystallography are employed to acquire such information at high resolution. However, application of these methods is limited because they are extremely demanding for protein purity and quantity, or protein size and crystalizability. Chemical cross-linking in combination with mass spectrometry (CXMS) is a novel technology developed over the past decade that complements the traditional methods with its many advantages such as high throughput, low cost and relaxed sample requirements. Here we review the various aspects of CXMS, including separation and enrichment techniques, characteristics of common cross-linkers, development of database-search tools for CXMS data, algorithms for validation of results, and application. The future development of CXMS is also discussed.

Abstract:Osteocyte is the most important functional cell type in osseous tissue, which is vital in response to mechanical stimuli, bone remodeling, mineral metabolism and homeostasis. Lacuna-canalicular networks serve as the fundamental structural microenvironment for osteocyte residing in and functioning. The rigid bone matrix mainly composed of type-Ⅰ collagen and hydroxyapatite provides the biochemical microenvironment for osteocyte adhesion and interaction with other cells and extracellular matrix. It is also accepted that the surrounding mechanical microenvironment is vital for osteocyte. Furthermore, due to the sensitive dependence on the surrounding environment, osteocyte will be greatly influenced by the changes of surrounding microstructures, biochemical components and mechanical stimulations. The microenvironment surrounding osteocyte is therefore of crucial importance to both elucidating the mechanism of mechanical perception and discovery of new biological functions. However, the complicated lacuna-canalicular network surrounded by rigid bone matrix makes the study of osteocyte in vivo technically difficult. It can be of great significance to reconstruct bionic microenvironment in vitro for further revealing the functions of osteocyte. The systematic introduction of structural, biochemical and mechanical microenvironment surrounding osteocyte was presented here, and the recent technologies for simulation of bionic microenvironment in vitro were also reviewed. This review will provide a useful reference for researchers who are interested in osteology, tissue engineering and regenerative medicine.

Abstract:The kinetics of Ca²⁺ triggered synaptic vesicle fusion contains synchronous and asynchronous components. At the Calyx of Held synapse, synaptotagmin-2 has been identified as the Ca² sensor protein for synchronous vesicle release while the Ca² sensor for asynchronous vesicle release remains unknown. Sr² is a divalent cation similar in size to Ca² and often substituted for Ca² as a tool in studying asynchronous vesicle release. However, the mechanism of Sr² triggering vesicle release is still under debate. We studied the spontaneous and evoked vesicle release at the wild type and synaptotagmin-2 knock-out calyceal synapses in the existence of extracellular Sr² and found synaptotagmin-2 only mediates the Sr² triggered synchronous vesicle release but not the asynchronous and spontaneous release; the unknown divalent sensor for asynchronous vesicle release has high-affinity to Sr² and mediates the spontaneous vesicle release. Our study provided a new insight into exploring the divalent sensor in triggering asynchronous synaptic vesicle release.

Abstract:The insulin-dependent uptake of glucose by adipose and muscle tissues is accomplished through the regulated vesicle trafficking of the GLUT4 glucose transporter to the plasma membrane. The Sec1p homologue Munc18c is believed to play a central role in the docking of GLUT4 vesicles by controlling SNARE complex assembly. In the present study we have examined the function of SM proteins in insulin-stimulated GLUT4 trafficking in adipocytes. Syntaxin4 at the plasma membrane is not dependent on the presence of Munc18c. We found that absence of Munc18c did not affect GLUT4 externalization at the plasma membrane and GLUT4 trafficking was normal in the absence of Munc18c and functional Syntaxin2, known to be associated with Munc18b. Syntaxin4 demonstrates a robust interaction with Munc18c but not either Munc18a or Munc18b in 3T3-L1 adipocytes. However, Munc18a and Munc18b exhibited weak interaction with Syntaxin4 in the background of absence of Munc18c. These data suggest that Syntaxin4 may play an important role in insulin-stimulated GLUT4 trafficking and its interaction with SM proteins are complementary.

Abstract:ECM stiffness has profound effects on cell migration, cell cycle, proliferation, differentiation, especially stem cell fate dictation. The underlying mechanism has not been elucidated so far. As a cell-adhesive protein, integrin is known as a mechanosensor positioned at the beginning of mechanotransduction. Our previous work demonstrates that ECM stiffness regulates the activity level and subcellular distribution of β1 integrin. However, the mechanism of β1 integrin activity regulation by ECM stiffness is still unclear. In the present paper, by using a monoclonal antibody specifically recognizing the active conformation of β1 integrin, we observed that soft ECM significantly enhanced the level of active β1 integrin in comparison of stiff ECM. Meanwhile, the activity of FAK-Raf-MEK-ERK was markedly promoted by stiff ECM. The inhibition of FAK-Raf pathway rescued the low activity level of β1 integrin on stiff ECM, indicating that stiff ECM may inhibits β1 integrin activity by FAK-Raf pathway in a negative feedback manner. In summary, we found a mechanism of the regulation of β1 integrin activity and downstream signal by ECM stiffness and provide a cue for understanding the mechanosensing of ECM stiffness by cells.

Abstract:We previously mapped a QTL significantly influencing the abdominal fat weight and the percentage of abdominal fat of chicken on chicken chromosome 7, and IGFBP2 is the only known gene located within this QTL region. We further found that the 1196C>A, a single nucleotide polymorphism (SNP) in 3'UTR of Chicken IGFBP2 gene, is significantly associated with chicken abdominal fat weight and percentage of abdominal fat. Bioinformatics analysis suggests that this SNP is located at a potential binding site of gga-miR-456-3p. We hypothesized that IGFBP2 may be a target gene of gga-miR-456-3p, and this SNP may affect the gga-miR-456-3p-mediated downregulation of IGFBP2. To test and decipher this hypothesis, in the present study we constructed the 3′UTR reporter vectors for the two individual alleles of the IGFBP2 SNP (1196C>A), and compared the reporter activity between the two individual alleles in both DF1 cells and chicken preadipocytes. Using the miRNA mimics and inhibitor of gga-miR-456-3p, we further assessed the effects of gga-miR-456-3p on the reporter activity of the two individual alleles of the IGFBP2 SNP and the endogenous IGFBP2 expression at mRNA and protein levels in DF1 cells. The reporter assay showed that in both DF1 cells and chicken preadipocytes, allele A had higher reporter activities at protein and mRNA levels than allele C, indicating that this SNP is a functional SNP. Further studies demonstrated that in DF1 cells, gga-miR-456-3p mimcs and inhibitor had effect on allele C reporter activity, but not on allele A reporter activity. Quantitative real-time RT-PCR and Western blot analyses showed that gga-miR-456-3p mimcs and inhibitor regulated the endogenous expression of chicken IGFBP2 gene at mRNA and protein levels. Taken together, our results demonstrated that IGFBP2 gene is a target gene of gga-miR-456-3p, and the SNP 1196C> A is a functional SNP. Our findings are of great significance to the marker-assisted selection for lean chicken and clarification of gene function and regulation of chicken IGFBP2 gene in chicken adipose development.

Abstract:Glycan-binding protein play important biological roles in biological processes. We use carbohydrate microarray to study the alteration of GBP in hepatocellular carcinoma cell line HepG2 and L02. Carbohydrate histochemistry was used to further validate the GBP and assess the distribution. As a result, 8 carbohydrate probes (e. g. SL, LNT, and GalNAc) showed increased signal while 5 carbohydrate probes (e. g. Man, Man-9-Glycan, and Xyl) showed decreased signal in HepG2 compared with L02 cell line. Meanwhile, GalNAc staining showed moderate binding to the cytoplasma membrane, central cytoplasm, and perinuclear cytoplasm in the L02, and the binding intensified in the same regions of the HepG2. NeuAc staining showed moderate binding to the cytoplasma membrane, and perinuclear cytoplasm in the HepG2, and the binding intensified in the same regions of the L02. In conclusion, the precision alteration of GBP related to HepG2 may provide useful information to find new molecular mechanism of hepatocellular carcinoma and antitumor therapeutic strategies.

Abstract:Advances in sequencing technology have led to a sharp decrease in the cost and rapid increase in the speed of sequencing an entire human genome. It has become one of the most important issues to analyze and visualize genome sequence in life science field. Genome browser technology plays important roles in analyzing genome sequence, interpreting genetic codes, studying complex diseases and so on. In this paper, we review the nine major genome browser technologies and analyze their characteristics in visualized content, visible form, software system architecture and so on. Finally, we discuss the challenges that genome browser faced.

Abstract:GRP75 of CHO Cells Responds to Ribosylation