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Objective To establish a THP-1 macrophage model infected by Mycobacterium smegmatis expressing green fluorescent protein (GFP), to quickly locate and visually detect Mycobacterium smegmatis, and to provide a tracer tool to identify the pathogenesis of tuberculosis and develop new tuberculosis vaccines. Methods The enhanced green fluorescent protein (EGFP) gene sequence was amplified by PCR using pEGFP-N1 plasmid as a template to obtain the coding gene of EGFP, and the amplified product was cloned into the vector pALACE to establish the recombinant plasmid pALACE-EGFP. Electroporation transformed the pALACE-EGFP into Mycobacterium smegmatis, and recombinant Mycobacterium smegmatis clones were screened by hygromycin resistance. After expanded culture, the smears were observed by fluorescence microscopy. The THP-1 macrophages were infected with recombinant Mycobacterium smegmatis, and the expression of EGFP was observed. Results The recombinant plasmid pALACE-EGFP was constructed appropriately. The recombinant Mycobacterium smegmatis was observed under fluorescence microscope. And it was confirmed that EGFP was expressed in recombinant Mycobacterium smegmatis, and THP-1 macrophages emitted green fluorescence after infection. Conclusion The successful establishment of recombinant Mycobacterium smegmatis expressing EGFP protein provides insights for investigating infection and pathogenesis of Mycobacterium tuberculosis.

Improving the transdermal absorption of weakly soluble drugs for topical use can help to prevent and treat skin photoaging. Nanocrystals of 18β-glycyrrhetinic acid (i.e., NGAs) prepared by high-pressure homogenization and amphiphilic chitosan (ACS) were used to form ANGA composites by electrostatic adsorption, and the optimal ratio of NGA to ACS was 10:1. Dynamic light scattering analysis and zeta potential analysis were used to evaluate the nanocomposites’ suspension, and the results showed that mean particle size was 318.8 ± 5.4 nm and the zeta potential was 30.88 ± 1.4 mV after autoclaving (121 °C, 30 min). The results of CCK-8 showed that the half-maximal inhibitory concentration (IC50) of ANGAs (71.9 μg/mL) was higher than that of NGAs (51.6 μg/mL), indicating that the cytotoxicity of ANGAs was weaker than that of NGAs at 24 h. After the composite had been prepared as a hydrogel, the vertical diffusion (Franz) cells were used to investigate skin permeability in vitro, and it was shown that the cumulative permeability of the ANGA hydrogel increased from 56.5 ± 1.4% to 75.3 ± 1.8%. The efficacy of the ANGA hydrogel against skin photoaging was studied by constructing a photoaging animal model under ultraviolet (UV) irradiation and staining. The ANGA hydrogel improved the photoaging characteristics of UV-induced mouse skin significantly, improved structural changes (e.g., breakage and clumping of collagen and elastic fibers in the dermis) significantly, and improved skin elasticity, while it inhibited the abnormal expression of matrix metalloproteinase (MMP)-1 and MMP-3 significantly, thereby reducing the damage caused by UV irradiation to the collagen-fiber structure. These results indicated that the NGAs could enhance the local penetration of GA into the skin and significantly improve the photoaging of mouse skin. The ANGA hydrogel could be used to counteract skin photoaging.

Objective To identify the effects of interleukin-6 (IL-6) on astrocytes activation, and the regulation of the expression of inwardly rectifying potassium 4.1 (Kir4.1) channels in astrocytes. Methods Astrocytes were separated from the cerebral cortex of newborn SD rats, and cultured in the presence of IL-6 or combined with interleukin-6 receptor antagonist (IL-6Ra). CCK-8 assay was performed to measure cell viability. The expression level of Kir4.1 channels in astrocytes was measured using quantitative real-time PCR and Western blot analysis. Results IL-6 promoted the proliferation of astrocytes in a dose- (0-30 ng/mL) and time- (0-24 hours) dependent manner. After astrocytes were treated with IL-6 (30 ng/mL) for 24 hours, the levels of Kir4.1 mRNA and protein decreased significantly, and this down-regulation could be attenuated by IL-6Ra. Conclusion IL-6 promotes the activation of astrocyte and down-regulation of the expression of Kir4.1 channel.

Phlomis medicinalis Diels, an important perennial herbal plant unique to the Qinghai-Tibet Plateau, is often used as Tibetan Materia Medicine Radix Phlomii for the treatment of cold, cough, and convergence trauma. In order to efficiently extract the iridoid glycosides from P. medicinalis, an ultrasound-assisted deep eutectic solvent extraction technique was employed. The main parameters influencing the extraction process were studied through single-factor tests and the extraction was optimized by using response surface methodology. The hemostasis activity of total iridoid glycosides (TIG) from P. medicinalis was evaluated in vitro and in mice. The optimization results revealed that the optimal process parameters were liquid-solid ratio 20 : 1, choline chloride-lactic acid concentration 79 %, and sonication time 34 min, under which a TIG extraction yield of 20.73 % was obtained. Meanwhile, high-performance liquid chromatography-photodiode array/mass spectrometry (HPLC-PDA/MS) was employed to characterize the optimized extract and indicated that TIG from P. medicinalis mainly consisted of sixteen reported iridoid glycosides with a total content of 91.22 %. The experimental results in vivo and in vitro indicated that TIG from P. medicinalis had strong hemostasis activities, which may be achieved by increasing the fibrinogen levels. Therefore, the ultrasound-assisted deep eutectic solvent extraction is an effective method to extract iridoid glycosides from P. medicinalis and they will be promising candidates to be developed for medical hemostasis agents.

Chitosan-based thermosensitive hydrogels have been widely used in drug delivery and tissue engineering, but their poor bioactivity has limited their further applications. Integral active oyster peptide microspheres (OPM) with an average particle diameter of 3.9 μm were prepared with high encapsulation efficiency (72.8%) and loading capacity (11.9%), exhibiting desirable sustained release effects. Using catechol functionalized chitosan (CS-C) as the polymeric matrix, OPM as the filler, and β-sodium glycerophosphate (β-GP) as a thermal sensitizer, the thermosensitive hydrogel CS-C/OPM/β-GP was prepared. Besides, the application of the hydrogel on wound healing was studied, and its biosafety was evaluated. The results of cell migration in vitro showed that the cell migration rate of CS-C/OPM/β-GP reached 97.47 ± 5.41% within 48 h, indicating that the hydrogel accelerated the migration of L929 cells. As demonstrated in the mouse skin wound experiment, CS-C/OPM/β-GP hydrogel not only inhibited the aggregation of diversified inflammatory cells and accelerated the generation of collagen fibers and new blood vessels of the wound, but also enhanced the synthesis of total protein (TP) in granulation tissue, and up-regulated the expression of Ki-67 and VEGF in the injury, thereby achieving fast wound healing. Safety evaluation results showed that CS-C/OPM/β-GP hydrogel was not cytotoxic to L929 cells, and the hemolysis ratio was less than 5% within 1 mg/mL. In conclusion, CS-C/OPM/β-GP hydrogel is expected as a promising medical dressing for wound healing.

Long-term alcohol intake or drinking large quantities of alcohol at one time can cause organ damage, which in turn can lead to chronic diseases. It is of important clinical and social significance to find effective approaches for the prevention and treatment of alcohol-induced diseases. In this paper, sulfhydryl functionalized chitosan (chitosan-N-acetyl-L-cysteine, CS-NAC) and sodium alginate (SA) were used as the matrix materials to contain tilapia peptide (TP), and a gastric acid-response hydrogel (CS-NAC/SA/TP) was prepared. Taking the ethanol adsorption rate as the response index, based on the results of the single factor test, the preparation process of CS-NAC/SA/TP was optimized through the Box–Behnken design. The swelling and antioxidant properties of CS-NAC/SA/TP were tested in vitro, and the protective effects on alcohol-induced acute liver injury and chronic brain injury were assessed in vivo. Structural characterization showed that CS-NAC/SA/TP was successfully prepared. Under the optimal conditions (SA concentration of 1%, MCS-NAC/MCaCO3 of 1 : 1, MSA/MCS-NAC(CaCO3) of 15 : 1), the prepared CS-NAC/SA/TP had a porous structure, a swelling ratio of 2350%, an ethanol adsorption rate of 56.23% and strong antioxidant capacities in vitro. Animal experiment results demonstrated that CS-NAC/SA/TP effectively reduced liver and brain injuries in mice caused by alcoholism. Summarily, these findings indicate that CS-NAC/SA/TP has potential applications in preventing alcohol-induced liver and brain injuries.

Based on Stephen Paget's well-established theory, both cell-autonomous and non-cell-autonomous mechanisms are crucial for metastasis. Although the mitochondrial calcium uniporter (MCU) has been suggested to be involved in breast cancer (BC) progression via cell-autonomous mechanisms, whether it assists the metastasis of BC cells through non-cell-autonomous mechanisms remains unclear. This study aimed to demonstrate that the MCU regulates BC metastatic colonization via non-cell-autonomous mechanisms. The results suggested that extracellular vesicles (EVs) derived from MCU-downregulated MDA-MB-231 cells suppressed angiogenesis in the metastatic niche in a nude mouse model, thereby hindering the colonization of BC cells. Mechanistically, we revealed that the MCU negatively correlated with miR-4488 in EVs derived from BC cells. Significantly, miR-4488 was determined to suppress angiogenesis of vascular endothelial cells by directly targeting angiogenic CX3CL1. Furthermore, we identified miR-4488 as being significantly downregulated in serum EVs from patients with triple-negative BC. Hence, this study suggests that MCU-dependent negative sorting of miR-4488 to EVs enhances angiogenesis in the metastatic niche and, thus, favors the metastatic colonization of BC cells.

Finger mobility plays a crucial role in everyday lives and is especially a leading indicator during hand rehabilitation and assistance tasks. In this study, a soft sensor-based three-dimensional (3-D) finger motion measurement system is proposed. Specifically, we report the use of a soft rope-embedded microfluidic sensor with high sensitivity and responsivity for strain sensing in the proposed system. These microfluidic strain sensors are proved to have the advantages of being easy to fabricate, chemically inert, with low hysteresis and good flexibility in their output signals. Besides, kinematic modeling of different finger joints are also provided to investigate the reasonable sensor locations for obtaining accurate measurements. Algorithms to identify the varied non-coplanar motions (abduction/adduction and flexion/extension joint angles) are presented by means of tracking the locations of the first metacarpal bone in multiple thumb postures. As a demonstration of their potential, the rope-embedded strain sensing units were used as curvature sensors mounted on the dorsum of target joints. The soft sensor-based 3-D finger motion measurement system and algorithms are experimentally verified by comparison with a camera-based motion capture device. Placing sensors at the optimal locations, joint angle measurement can reach up

Objective To investigate the effect of insulin-like growth factor 1 (IGF-1) on the phagocytic activity of mouse BV-2 microglial cells. Methods Western blotting was performed to detect the protein levels of IGF-1 and IGF-1 receptor (IGF-1R) in the murine brain after the establishment of acute central nervous system inflammation models by intraperitoneal lipopolysaccharide (LPS) injection (10 mg/kg). The protein level of IGF-1R on BV-2 microglial cells that had been stimulated by 500 ng/mL LPS for 4, 12 and 24 hours was measured by Western blotting. To assess the phagocytic activity of microglial cells in response to IGF-1, BV-2 microglial cells were stimulated by IGF-1 at different concentrations for 24 hours after pretreated with or without wortmannin (PI3K/AKT signaling pathway blocker), and then incubated with fluorescent microbeads for 2 hours followed by measurement of phagocytosis of the fluorescent microbeads by flow cytometry. After treatment of IGF-1 (50 ng/mL), p-AKT and AKT signaling pathways in the BV-2 microglial cells were detected by Western blotting. Results Intraperitoneal LPS injection caused increased levels of IGF-1 and IGF-1R in the mouse brain. LPS upregulated the protein expression of IGF-1R on BV-2 microglial cells. The activity of BV-2 microglial cells to phagocytose fluorescent microbeads gradually increased with IGF-1 concentration rising and peaked in the IGF-1 treatment at 50 ng/mL, and gradually decreased thereafter. And IGF-1 induced the phosphorylation of AKT in BV-2 microglial cells. However, after the PI3K/AKT signaling pathway was blocked via wortmannin, the effect of IGF-1 on the activity of BV-2 microglial cells to phagocytose fluorescent microbeads was significantly alleviated. Conclusion IGF-1 can promote phagocytic activity of BV-2 cells via activating PI3K/AKT signaling pathway, which suggests a potential role of IGF-1 in regulating the cerebral inflammation.

18β-Glycyrrhetinic acid (GA) is often topically applied in clinical treatment of inflammatory skin diseases. However, GA has poor solubility in water, which results in poor skin permeability and low bioavailability. Nanocrystallization of drugs can enhance their permeability and improve bioavailability. We prepared GA nanocrystals (Nano GA) by high-pressure homogenization. These nanocrystals were characterized by photon correlation spectroscopy, scanning electron microscopy, thermogravimetric analysis, and X-ray diffractometry. The ability of Nano GA to improve dermal permeability was investigated ex vivo using Franz diffusion vertical cells and mouse skin. The topical anti-inflammatory activity of Nano GA was assessed in vivo by a 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced model in mouse ears. The average particle size of a GA nanocrystalline suspension was 288.6 ± 7.3 nm, with a narrow particle-size distribution (polydispersity index ∼0.13 ± 0.10), and the particle size of the lyophilized powder increased (552.0 ± 9.8 nm). After nanocrystallization, the thermal stability and crystallinity decreased but solubility increased significantly. Nano GA showed higher dermal permeability than Coarse GA. Macroscopic and staining-based observations of mouse ears and the levels of proinflammatory factors and myeloperoxidase revealed that the Nano GA hydrogel exhibited better anti-edema ability and more strongly inhibited inflammation development than the Coarse GA hydrogel and indomethacin hydrogel (positive drug). These results suggest that Nano GA could be an efficacious topical therapeutic agent for skin inflammation.

Uncontrolled massive hemorrhage is one of the principal causes of death in trauma emergencies. By using catechol-modified chitosan (CS-C) as the matrix material and &beta, glycerol phosphate (&beta, GP) as a thermo-sensitive agent, chitosan-based thermo-sensitive hydrogel loading oyster peptides (CS-C/OP/&beta, GP) were prepared at physiological temperature. The hemostatic performance of CS-C/OP/&beta, GP hydrogel was tested in vivo and in vitro, and its biological safety was evaluated. The results showed that the in vitro coagulation time and blood coagulation index of CS-C/OP/&beta, GP hydrogel were better than those of a commercial gelatin sponge. Notably, compared with the gelatin sponge, CS-C/OP/&beta, GP hydrogel showed that the platelet adhesion and erythrocyte adsorption rates were 38.98% and 95.87% higher, respectively. Additionally, the hemostasis time in mouse liver injury was shortened by 19.5%, and the mass of blood loss in the mouse tail amputation model was reduced by 18.9%. The safety evaluation results demonstrated that CS-C/OP/&beta, GP had no cytotoxicity to L929 cells, and the hemolysis rates were less than 5% within 1 mg/mL, suggesting good biocompatibility. In conclusion, our results indicate that CS-C/OP/&beta, GP is expected to be a promising dressing in the field of medical hemostasis.

Long-term excessive alcohol intake can easily lead to gastritis, gastric ulcer, and gastric bleeding. In this paper, the gastric acid-responsive hydrogel of CS-NAC/alginate/tilapia collagen peptide (CS-NAC/ALG/TCP) was developed. Its structure and properties were determined. The alcohol-induced gastric mucosal injury models in mice were established to evaluate the protective effects of CS-NAC/ALG/TCP. The results showed that CS-NAC/ALG/TCP was successfully fabricated, and it showed a sustained release of TCP, strong mucoadhesion, and excellent biodegradability in vitro. In the animal experiments, CS-NAC/ALG/TCP improved the oxidative stress status of the gastric mucosa by increasing the levels of SOD, GSH, and CAT in tissues. It also down-regulated the expression of MPO, TNF-α, IL-1β, and IL-6, and increased the production of gastric protective factors such as PGE2 and NO in mouse stomach, thereby reducing the alcohol-induced inflammation and protecting the gastric mucosal injury. Besides, CS-NAC/ALG/TCP can also increase the activities of alcohol metabolism enzymes to improve alcohol metabolism, thereby reducing alcoholic damage. In conclusion, CS-NAC/ALG/TCP is a promising candidate for the treatment of alcohol-induced gastric injury.

The safety of human-machine interaction is very important in intelligent manufacturing, and the main purpose of ensuring safety is to enable the robot to receive information about the surrounding environment through proximity sensors, which can achieve good interaction between the machine and humans. In view of the current shortcomings of electronic skins, such as poor flexibility, lack of proximity sensing system, and the immeasurable magnitude of force after contact, we design a composite sensor that can realize proximity and small force haptics in this paper. We use flexible materials such as copper foil, polyimide, polyurethane and a new type of semiconductor material based on super capacitor and design a double-layer pressure-capacitance sensing structure based on the principle of human induced capacitance, variable dielectric constant and variable pitch capacitance. And we use STM32, FDC2214 chip to build a complete information acquisition and processing system.

Uncontrolled bleeding has always been a sudden accident, which is the main cause of casualties in war trauma, emergency events and surgical operations. Rapid hemostatic materials can effectively reduce casualties and save lives. In this paper, marine collagen peptide grafted carboxymethyl chitosan (CMCS-MCP) was synthesized by 1-ethyl-(dimethylaminopropyl) carbodiimide (EDC)-mediated coupling reaction. To obtain CMCS-MCP conjugates with different degrees of substitution (DS), the reaction conditions were investigated by single-factor tests and optimized by response surface methodology. And the sponges of CMCS-MCP were prepared by freeze-thaw cycling and freeze-drying and characterized by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), and X-ray diffraction (XRD). To evaluate the hemostatic properties of CMCS-MCP sponges, in vitro and in vivo hemostasis tests were carried out. The results showed that the optimum preparation conditions were the mass ratio of MCP to CMCS (MMCP/MCMCS) 6:1, reaction temperature 41 °C, and reaction time 16 h. And under which the DS of 58.86% was obtained. Structure analysis showed that MCP had been successfully grafted onto the CMCS molecular chain, and the CMCS-MCP sponges were of high porosity. In vitro and in vivo hemostasis tests showed that the CMCS-MCP sponges had significant procoagulant activities, especially the one with high DS of 58.86%. The hemostasis mechanism may be that the synergistic effects of MCP and CMCS accelerated coagulation through multiple approaches. The CMCS-MCP sponges give a new insight into biomedical hemostasis materials.

Lung metastasis is a primary obstacle in the clinical treatment of metastatic breast cancer. Most patients with lung metastasis eventually die of recurrence. Recurrence may be related to self-seeding, which occurs when circulating tumor cells re-seed into the tumors they originated from (metastasis or carcinoma in situ). Tumor-derived exosomes have been intensively revealed to promote the progression of various cancers. However, whether tumor-derived exosomes play roles in tumor self-seeding has not yet been identified. By establishing a self-seeding nude mouse model, we found that exosomes derived from MDA231-LM2 cells (subpopulations of breast cancer lung metastasis) potentiate the growth of MDA-MB-231 xenografts. More importantly, laser confocal microscopy and flow cytometry results identified that MDA231-LM2-secreted exosomes promote the seeding of MDA231-LM2 cells into MDA-MB-231 xenografts. These findings suggest MDA231-LM2-secreted exosomes as a promising target to treat breast cancer lung metastasis.

In this paper, a novel flexible tactile sensor array based on conductive rubber is presented. Compared with other research results, we use the materials with complete flexibility to fabricate the soft sensors. The sensing prototype includes polydimethylsiloxane (PDMS) as the sealing layers and conductive rubber as sensing layer. This proposed tactile sensor can measure the contact pressure under wrapping one curved surface. In our study, we developed a skin-like tactile sensor array from three aspects: design procedures, fabrication techniques and test results via taking one 8×8 sensor array for example, which is a sandwich structure and the sensing material was laminated into a thin encapsulation film with 2mm in thickness. Furthermore, the sensor overcomes the fabrication complexity, chemical unsafety and environmental instability problems to some extent. In the end, the final experimental demonstration verify the feasibility of the sensor array with discrete sensing elements in measuring the tactile force via matlab.

Anodic TiO2 nanotube arrays (ATNAs) have been one of the promising electrode materials for supercapacitors due to good chemical stability, large surface area and simple fabrication method. However, the specific capacitance of ATNAs is generally low. Here, we focus on the hydrothermal treatments of ATNAs in (NH4)2TiF6 dilute solutions. After the (NH4)2TiF6 hydrothermal treatments, the smooth wall of ATNAs became rough, a layer of nanoparticles were generated on the surface of nanotubes concurrently. The specific capacitance of the hydrothermally treated ATNAs increased remarkably due to the special nanostructure composed of rough tube walls and the nanoparticles embedded on the tube walls. After the hydrothermal treatment in 0.01 M (NH4)2TiF6 aqueous solution at 120 °C for 45 min, the ATNAs exhibit a specific capacitance of 31.12 mF cm−2, about a 3.1-fold improvement over the untreated ones.

Special nanostructures of porous anodic alumina (PAA), such as the petal-like micropattern and nanofibers of alumina, cannot be explained by the classical field-assisted dissolution and dynamic equilibrium models. Here, by combination of oxygen bubble mould and plastic flow model, a reasonable explanation for these special nanostructures is proposed. The petal-like micropatterns of PAA and nanofibers of alumina are all clarified by the oxygen bubble mould and the plastic flow model. The self-ordering nanochannels of PAA are attributed to the oxide plastic flow around a gas bubble mould at each pore bottom. The petal-like micropatterns and nanofibers of alumina are attributed to chemical corrosion enhanced by the micro-gas-flow and micro-liquid-flow.

Previous studies suggested that high-field anodization of aluminum can be realized given that aluminum is anodized at a high voltage just below the breakdown value. However, increasing the applied voltage cannot guarantee the enhancement in electric field strength across the barrier layer of porous anodic alumina (PAA) due to a concurrent increase of the barrier-layer thickness. Here, we report comparative studies of aluminum anodization in a highly concentrated (0.75 M) and a dilute (0.1 M) oxalic acid solution. Special attention is given to the field strength during anodization. To calculate the field strengths, the barrier-layer thickness of PAA was determined by a re-anodizing technique. Electrochemical impedance spectra (EIS) were also used to measure the barrier-layer thickness to improve the reliability of the field strength measurements. Both routes can yield the same conclusion: the barrier-layer thickness increases linearly with the applied voltage. For 0.75 M oxalic acid electrolyte, the resulting regression line has a positive intercept. In this instance the field strengths can be enhanced by increasing the applied voltage. Conversely, for 0.1 M oxalic acid solution, the regression line has a negative intercept and the field strengths decrease with the applied voltage. There are different linear dependences between the barrier-layer thickness and the applied voltage that determine the change in the field strength during anodization. In the highly concentrated oxalic acid electrolyte, the high-field anodization of aluminum can be realized by enhancing the applied voltage, but cannot in the dilute acid solution. Moreover, the ordering qualities of PAA films increase with increasing field strength instead of the applied voltage. The present results may provide a decisive step towards a thorough understanding of the PAA film.

Burns are physically debilitating and potentially fatal injuries. Two marine biomaterials, carboxymethyl chitosan (CMC) and collagen peptides (COP), have emerged as promising burn dressings. In this paper, sponges of carboxymethyl chitosan grafted with collagen peptide (CMC&ndash, COP) were prepared by covalent coupling and freeze drying. Scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy were then used to characterize the prepared sponges. To evaluate the wound healing activity of the CMC&ndash, COP sponges, in vitro tests including cell viability scratch wound healing and scald wound healing experiments were performed in rabbits. Appearance studies revealed the porous nature of sponges and FTIR spectroscopy demonstrated the successful incorporation of COP into CMC. The in vitro scratch assay showed that treatment with CMC&ndash, COP sponges (at 100 &mu, g/mL) had significant effects on scratch closure. For burn wounds treated with CMC&ndash, COP, regeneration of the epidermis and collagen fiber deposition was observed on day 7, with complete healing of the epidermis and wound on days 14 and 21, respectively. Based on the pathological examination by hematoxylin and eosinstaining, the CMC&ndash, COP group demonstrated pronounced wound healing efficiencies. These results confirmed that the CMC&ndash, COP treatment enhanced cell migration and promoted skin regeneration, thereby highlighting the potential application of these sponges in burn care.

Background Stromal interaction molecule (STIM) 2 is a key calcium-sensing molecule that regulates the stabilization of calcium ions (Ca2+) and therefore regulates downstream Ca2+-associated signaling and cellular events. We hypothesized that STIM2 regulates epithelial-mesenchymal transition (EMT) to promote breast cancer metastasis. Methods We determined the effects of gain, loss, and rescue of STIM2 on cellular motility, levels of EMT-related proteins, and secretion of transforming growth factor-β (TGF-β). We also conducted bioinformatics analyses and in vivo assessments of breast cancer growth and metastasis using xenograft models. Results We found a significant association between STIM2 overexpression and metastatic breast cancer. STIM2 overexpression activated the nuclear factor of activated T cells 1 (NFAT1) and TGF-β signaling. Knockdown of STIM2 inhibited the motility of breast cancer cells by inhibiting EMT via specific suppression of NFAT1 and inhibited mammary tumor metastasis in mice. In contrast, STIM2 overexpression promoted metastasis. These findings were validated in human tissue arrays of 340 breast cancer samples for STIM2. Conclusion Taken together, our results demonstrated that STIM2 specifically regulates NFAT1, which in turn regulates the expression and secretion of TGF-β1 to promote EMT in vitro and in vivo, leading to metastasis of breast cancer. Electronic supplementary material The online version of this article (10.1186/s13058-019-1185-1) contains supplementary material, which is available to authorized users.

FAM64A is a mitotic regulator promoting cell metaphase–anaphase transition, and it is frequently reported to be highly expressed in cancer cells. However, the role of FAM64A in human breast cancer (BrC) is poorly studied. The expression of FAM64A mRNA in BrC samples was determined by RT-qPCR assay and TCGA database mining. Kaplan–Meier plotter was used to analyze whether FAM64A expression impacted prognosis. Then, the expression of FAM64A was silenced using RNA interference. Cell-counting assay, colony formation assay and flow cytometry assay were conducted to detect proliferation; transwell migration assay, EMT-related proteins expression (E-cadherin, N-cadherin and vimentin), and EMT-related transcription factors mRNA expression (Snail, Twist, Slug) were conducted to evaluate the migration ability. FAM64A was highly expressed in human BrC samples, which was negatively associated with poor survival time. Analysis of FAM64A expression in BrC cell lines demonstrated that the expression of FAM64A was significantly correlated with the proliferation rate and migration ability of BrC cells. Indeed, knockdown of FAM64A suppressed the proliferation of MDA-MB-231 and MCF-7 cells. Importantly, we also found that silencing of FAM64A inhibited the migration of BrC cells via impeding epithelial–mesenchymal transition. Our findings suggest that FAM64A plays an important role in the proliferation and migration of BrC cells, which might serve as a potential target for BrC treatment.

Online Supplementary Methods Section. (DOCX 20 kb)

IACUC Aprroval Document for Mouse Breast Cancer Xenograft Tumor Assessments. (PDF 643 kb)

IRB Approval Document for Human Breast Cancer Tissue Microarray Assessments. (PDF 2603 kb)