Template free preparation of graphene tubes from polyimide catalyzed by calcium carbonate.

This work reports a new means of preparing graphene tubes (GTs) without relying on chemical vapor deposition (CVD) and it's template-free. Surprisingly, we found that under the action of calcium oxide (CaO) and after 1500 °C heat treatment, a large amount of GTs grew on the surface of polyimide (PI). These nanotubes have a maximum diameter of about 600 nm and a length of up to millimeters, and some nanotubes even have a branching structure. We propose a simple, effective and green method which exhibits prospects for large-scale production of GTs using polymeric materials.

1α,25-Dihydroxyvitamin D3 Improves Follicular Development and Steroid Hormone Biosynthesis by Regulating Vitamin D Receptor in the Layers Model.

1α,25-Dihydroxyvitamin D3 (VitD3) is the active form of vitamin D, and it regulates gene expression and protein synthesis in mammalian follicle development. However, the function of VitD3 in the follicular development of layers remains unclear. This study investigated, through in vivo and in vitro experiments, the effects of VitD3 on follicle development and steroid hormone biosynthesis in young layers. In vivo, ninety 18-week-old Hy-Line Brown laying hens were randomly divided into three groups for different treatments of VitD3 (0, 10, and 100 µg/kg). VitD3 supplementation promoted follicle development, increasing the number of small yellow follicles (SYFs) and large yellow follicles (LYFs) and the thickness of the granulosa layer (GL) of SYFs. Transcriptome analysis revealed that VitD3 supplementation altered gene expression in the ovarian steroidogenesis, cholesterol metabolism, and glycerolipid metabolism signaling pathways. Steroid hormone-targeted metabolomics profiling identified 20 steroid hormones altered by VitD3 treatment, with 5 being significantly different among the groups. In vitro, it was found that VitD3 increased cell proliferation, promoted cell-cycle progression, regulated the expression of cell-cycle-related genes, and inhibited the apoptosis of granulosa cells from pre-hierarchical follicles (phGCs) and theca cells from prehierarchical follicles (phTCs). In addition, the steroid hormone biosynthesis-related genes, estradiol (E2) and progesterone (P4) concentrations, and vitamin D receptor (VDR) expression level was significantly altered by VitD3. Our findings identified that VitD3 altered the gene expression related to steroid metabolism and the production of testosterone, estradiol, and progesterone in the pre-hierarchical follicles (PHFs), resulting in positive effects on poultry follicular development.

Characterization and proteomics of chicken seminal plasma extracellular vesicles.

In mammals, seminal plasma extracellular vesicles (SPEVs) can regulate sperm motility and capacitation. The characteristics and functions of SPEVs in avians have been rarely reported. In this study, chicken SPEVs were isolated and characterized by transmission and scanning electron microscopy (TEM/SEM) and nanoparticle tracking analysis (NTA); furthermore, seven extracellular vesicle (EVs) marker proteins were detected by Western blot (WB). TEM revealed that chicken SPEVs had a classic bilayer membrane structure. NTA confirmed that the size of SPEVs was 30-250 nm, and concentration ranged from 8.0 E + 11-8.5 E + 11 particles/ml. There were 3073 SPEVs proteins identified by deep sequencing, including 2794 intracellular proteins and 279 extracellular proteins. The overlap rate of proteomes between chicken SPEVs and vesicles reported in the Vesiclepedia database reached 86%, and 360 new proteins that had not been reported by the ExoCarta and Vesiclepedia databases were identified in chicken SPEV proteomes. Gene Ontology (GO) analysis revealed that chicken SPEV proteins were mainly enriched in supplying energy and transporting protein. There were 4 IFT family proteins speculated to play an important role in sperm composition and function. Our data were compared with two previously published studies on the proteomics of chicken seminal plasma (SP) and hen uterine fluid, and some overlapping proteins described in chicken SPEVs had been identified in hen uterine fluid (545) and chicken SP (284). In conclusion, these findings will increase our understanding of the content and composition of proteome in SPEVs and provide new insights into the important role of the SPEV regulation in sperm functions.

BML-111, the lipoxin A4 agonist, modulates VEGF or CoCl2-induced migration, angiogenesis and permeability in tumor-derived endothelial cells.

Tumor angiogenesis plays a vital role in carcinogenesis, cancer progression, and metastasis. Lipoxin A4 (LXA4) is an endogenously-produced family of effective anti-inflammatory with a potent inhibitory effect on angiogenesis. However, BML-111, a LXA4 agonist, its governing tumor-derived endothelial cells (Td-EC) mechanisms remain unknown. In the present study, we utilized VEGF or CoCl2 to mimic tumor microenvironment in vitro to study the effect of BML-111 on angiogenesis and permeability of Td-EC, and preliminarily explore its specific mechanism. Data suggested that BML-111 inhibited viability, migration and angiogenesis in VEGF or CoCl2-treated Td-EC by modulating MMP2/9-TIMP1, and decreasing the production of HIF-1α and COX-2 level. In addition, we observed that BML-111 inhibited Td-EC permeability induced by VEGF or CoCl2, through the stabilization of VE-cadherin/ß-catenin-dependent adherens junctions and TRPC1 pathway. Nevertheless, these effects could be blocked by BOC-2 which was the specific inhibitor of FPR2/ALX (the receptor of LXA4).These results suggest that BML-111 may have inhibitory effects on VEGF or CoCl2-induced migration, angiogenesis and permeability in tumor-derived endothelial cells.

Comparative Analysis Among Different Species Reveals That the Androgen Receptor Regulates Chicken Follicle Selection Through Species-Specific Genes Related to Follicle Development.

The differences in reproductive processes at the molecular level between viviparous and oviparous animals are evident, and the site in the ovary that synthesizes sex hormones (androgens and oestrogens) and the trends for enriching sex hormones during follicle development in chickens are different from those in mammals, suggesting that the effect of sex hormones on follicle development in chickens is probably different from that in viviparous animals. To explore the specific role of androgen receptors (ARs) on chicken follicular development, we matched the correspondence of follicular development stages among chickens, humans, cows and identified chicken-specific genes related to follicle development (GAL-SPGs) by comparing follicle development-related genes and their biological functions among species (chickens, humans, and cows). A comparison of the core transcription factor regulatory network of granulosa cells (or ovaries) based on super-enhancers among species (chicken, human, and mouse) revealed that AR is a core transcriptional regulator specific to chickens. In vivo experiments showed that inhibition of AR significantly reduced the number of syf (selected stage follicles) in chickens and decreased the expression of GAL-SPGs in F5 follicles, while in vitro experiments showed that inhibition of AR expression in chicken granulosa cells (GCs) significantly down-regulated the expression levels of GAL-SPGs, indicating that AR could regulate follicle selection through chicken-specific genes related to follicle development. A comparison among species (77 vertebrates) of the conserved genomic regions, where chicken super-enhancers are located, revealed that the chicken AR super-enhancer region is conserved in birds, suggesting that the role of AR in follicle selection maybe widespread in birds. In summary, we found that AR can regulate follicle selection through chicken-specific genes related to follicle development, which also emphasizes the important role of AR in follicle selection in chickens and provides a new perspective for understanding the unique process of follicle development in chickens. Our study will contribute to the application of androgens to the control of egg production in chickens and suggests that researchers can delve into the mechanisms of follicle development in birds based on androgen/androgen receptors.

BML-111 inhibits EMT, migration and metastasis of TAMs-stimulated triple-negative breast cancer cells via ILK pathway.

Triple-negative breast cancer (TNBC) has a more aggressive phenotype and higher metastasis and recurrence rates than other breast cancer subtypes. The immune microenvironment and hypoxic microenvironment of breast cancer constitute the survival environment of cancer cells, which is an important environment to support cancer cells. LXA4 and its analog, BML-111 is an important regulator of inflammatory cytokines, which provides a possible way for the treatment of inflammatory-related tumors. Here, in the in vitro experiment, we showed that BML-111 could inhibit the EMT and migration of TAMs-stimulated TNBC by down-regulating ILK as well as p-Akt and p-GSK3ß. And it could prevent the formation of breast cancer cell clusters. In the in vivo experiment, BML-111 could inhibit the metastasis of 4T1 breast cancer cells. We also demonstrated that BML-111 could affect macrophages in tumor microenvironment to prevent metastasis. These results showed that BML-111 could be a possible candidate for breast cancer therapy by targeting ILK and TAMs.

SNORA71B promotes breast cancer cells across blood-brain barrier by inducing epithelial-mesenchymal transition.

BACKGROUND: Brain metastasis (BM) is a dreadful complication that significantly impacts the quality of life in breast cancer patients. A key process during brain metastasis is the migration of cancer cells across blood-brain barrier (BBB). However, the role of snoRNAs regulating BBB in BM is still unknown. METHODS: Here SNORic and GEO databases were used to identify differentially expressed snoRNAs between brain metastatic and non-metastatic breast cancer (BC) tissues. The effects of SNORA71B on the capacities of proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), and BBB invasion of BC cells were evaluated by CCK8, transwell, western blot, and BBB model, respectively. RESULTS: SNORA71B was highly expressed in high BM BC tissues and cells compared to low BM BC controls. Survival analysis revealed high expression of SNORA71B was significantly associated with poor PPS and OS in breast cancer patients. ROC curve showed that SNORA71B might act as biomarker for breast cancer. Moreover, SNORA71B significantly promoted proliferation, migration, and invasion of BC cells with different BM abilities. Importantly, SNORA71B promoted the EMT process of low BM BC cells. SNORA71B knockdown inhibited the high BM BC cells across BBB, while EMT activator dramatically abrogated this inhibited effect. CONCLUSIONS: In conclusion, SNORA71B promotes BC cells across the BBB partly via inducing EMT.

EMT related circular RNA expression profiles identify circSCYL2 as a novel molecule in breast tumor metastasis.

Substantial evidence indicates that circular RNAs (circRNAs) play vital roles in several diseases, especially in cancer development. However, the functions of circRNAs in breast cancer metastasis remain to be investigated. This study aimed to identify the key circRNAs involved in epithelial mesenchymal transition (EMT) of breast cancer and evaluated their molecular function and roles in pathways that may be associated with tumor metastasis. An EMT model was constructed by treating breast cancer cells MCF­7 and MDA­MB­231 with transforming growth factor­ß1. High­throughput RNA sequencing was used to identify the differentially expressed circRNAs in EMT and blank groups of two cells, and reverse transcription­quantitative PCR was used to validate the expression of circSCYL2 in human breast cancer tissues and cells. The effects of circSCYL2 on breast cancer cells were explored by transfecting with plasmids and the biological roles were assessed using transwell assays. EMT groups of breast cancer cells exhibited the characteristics of mesenchymal cells. Furthermore, the present study found that 7 circRNAs were significantly upregulated in both the MCF­7 EMT and MDA­MB­231 EMT groups, while 16 circRNAs were significantly downregulated. The current study identified that circSCYL2 was downregulated in breast cancer tissues and cell lines, and that circSCYL2 overexpression inhibited cell migration and invasion. This study provides expression profiles of circRNAs in EMT groups of breast cancer cells. circSCYL2, which is downregulated in breast cancer tissues and cells, may play an important role in breast cancer EMT progression.

Development and characterization of mammary intraductal (MIND) spontaneous metastasis models for triple-negative breast cancer in syngeneic mice.

Triple-negative breast cancer (TNBC) has a more aggressive phenotype and higher metastasis and recurrence rates than other breast cancer subtypes. TNBC currently lacks a transplantation model that is suitable for clinical simulations of the tumor microenvironment. Intraductal injection of tumor cells into the mammary duct could mimic the occurrence and development of breast cancer. Herein, we injected 4T1 cells into the mammary ducts of BALB/C mice to build a preclinical model of TNBC and optimized the related construction method to observe the occurrence and spontaneous metastasis of tumors. We compared the effects of different cell numbers on tumorigenesis rates, times to tumorigenesis, and metastases to determine the optimal number of cells for modelling. We demonstrated that 4T1-MIND model mice injected with 20,000 cells revealed a suitable tumor formation rate and time, thus indicating a potential treatment time window after distant metastasis. We also injected 20,000 cells directly into the breast fat pad or breast duct for parallel comparison. The results still showed that the 4T1-MIND model provides sufficient treatment time for lung metastases in mice and that it is a more reliable model for early tumor development. The 4T1-MIND model requires continuous improvement and optimization. A suitable and optimized model for translational research and studies on the microenvironment in TNBC should be developed.

Long noncoding RNA LINC00115 promotes breast cancer metastasis by inhibiting miR-7.

Breast cancer is the second leading cause of cancer-related deaths in women. The long noncoding RNA LINC00115 has been reported to be involved in the poor outcome of patients with breast cancer, but the biological function and underlying mechanism remain unclear. Here, we report that LINC00115 expression is increased in triple-negative breast cancer tissue compared with matched normal tissue, and LINC00115 knockdown suppresses breast cancer cell migration and invasion. Furthermore, we show that LINC00115 directly targets miR-7 and inhibits its expression. LINC00115 also reduces the expression of KLF4, which is a direct target of miR-7 and is involved in breast cancer metastasis. Together, our findings suggest that LINC00115 promotes breast cancer metastasis through modulating the expression of miR-7 and KLF4.

Interactions of Biocidal Polyhexamethylene Guanidine Hydrochloride and Its Analogs with POPC Model Membranes.

The bacterial membrane-targeted polyhexamethylene guanidine hydrochloride (PHGH) and its novel analog polyoctamethylene guanidine hydrochloride (POGH) had excellent antimicrobial activities against antibiotics-resistant bacteria. However, the biocompatibility aspects of PHGH and POGH on the phospholipid membrane of the eukaryotic cell have not yet been considered. Four chemically synthesized cationic oligoguanidine polymers containing alkyl group with different carbon chain lengths, including PHGH, POGH, and their two analogs, were used to determine their interactions with zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) phospholipids vesicles mimicking the eukaryotic cell membrane. Characterization was conducted by using bactericidal dynamics, hemolysis testing, calcein dye leakage, and isothermal titration calorimetry. Results showed that the gradually lengthened alkyl carbon chain of four oligoguanidine polymers increased the biocidal activity of the polymer, accompanied with the increased hemolytic activity, calcein dye leakage rate and the increased absolute value of the exothermic effect of polymer-POPC membrane interaction. The thermodynamic curve of the polymer-POPC membrane interaction exhibited a very weak exothermic effect and a poorly unsaturated titration curve, which indicated that four guanidine polymers had weak affinity for zwitterionic POPC vesicles. Generally, PHGH of four guanidine polymers had high biocidal activity and relatively high biocompatibility. This study emphasized that appropriate amphiphilicity balanced by the alkyl chain length, and the positive charge is important factor for the biocompatibility of cationic antimicrobial guanidine polymer. Both PHGH and POGH exhibited destructive power to phospholipid membrane of eukaryotic cell, which should be considered in their industry applications.