VEGF Research Articles

In-vivo oogenesis of oogonial and mesenchymal stem cells seeded in transplanted ovarian extracellular matrix

Objective (s)One way to overcome the recurrence of cancer cells following ovarian tissue transplantation is to use decellularized tissues as a scaffold that does not have any cellular components. These cell-free scaffolds can be seeded with different type of stem cells for ovarian restoration.Materials and methodsOSCs, PMSCs and BMSCs (oogonial, peritoneal and bone marrow mesenchymal stem cells, respectively) were seeded into human decellularized ovarian tissue as 4 groups: Scaffold + OSCs (SO), Scaffold + OSC + PMSCs (SOP), Scaffold + OSC + BMSCs (SOB) and Scaffold + OSC + PMSCs + BMSCs (SOPB). The produced grafts were transplanted into the sub-peritoneal space of ovariectomized NMRI mice as artificial ovary (AO). The expression of Vegf, CD34, Gdf9, Zp3, Ddx4, Amh and Lhr genes in AOs were measured by qRT-PCR. Also, histotechniques were considered to detect the anti GFP, PCNA, VEGF, GDF9, ZP3 and AMH proteins.ResultsH & E staining showed follicle-like structures in all groups; the number of these structures, in the SOP and SOB groups, were the highest. In SO group, differentiation ability to oocyte and granulosa cells was observed. Endothelial, oocyte, germ, and granulosa cell-like cells were specially seen in SOP and angiogenesis capability was more in SOB group. However, angiogenesis ability and differentiation to theca cell-like cells were more often in SOPB group. While none of the groups showed a significant difference in AMH level, estradiol levels were significantly higher in SOPB group.ConclusionIntegration of OSCs + PMSCs and those OSCs + BMSCs were more conducive to oogenesis.

Evaluation of collagen type I and III, TGF-β1, and VEGF gene expression in rat skin wound healing treated by alginate/chitosan hydrogel containing crocetin

Skin tissue engineering is one of the popular strategies are used to improve the wound healing process. In this study, different concentration of Crocetin (Crot) were loaded into a three-dimensional (3D) alginate (Alg) and chitosan (CS) hydrogel to improve wound healing process. After hydrogel preparation, it is characterized with different experiments such as morphology evolution, water absorption, cumulative release, weight loss, hemo- and cytocompatibility, cell viability, and antibacterial properties. Then, the therapeutic role of the prepared hydrogel was investigated in a full-thickness dermal wound in a rat model with analyzing RNA expression and histology. The results showed that fabricated hydrogels had sufficient porosity and the weight loss test verified the appropriate biodegradability of the prepared hydrogel (about 90% after 14 days) that can release enough of the Crot without any toxicity effect. In addition, the in vivo findings showed that the alginate/chitosan/crocetin 150 µM group had the highest wound closure percentage, re-epithelialization, and gene expression. Furthermore, rt-PCR revealed that 150 µM crocetin increased the expression of Collagen type I, III, VEGF, and TGF-β mRNA. Overall, this research indicates that 3D Alg/CS hydrogels containing 150 µM crocetin may be used in the clinic to treat skin injuries.

Effect of Atorvastatin on Angiogenesis-Related Genes VEGF-A, HGF and IGF-1 and the Modulation of PI3K/AKT/mTOR Transcripts in Bone-Marrow-Derived Mesenchymal Stem Cells.

Stem cell transplantation represents a unique therapeutic tool in tissue engineering and regenerative medicine. However, it was shown that the post-injection survival of stem cells is poor, warranting a more comprehensive understanding of activated regenerative pathways. Numerous studies indicate that statins improve the therapeutic efficacy of stem cells in regenerative medicine. In the present study, we investigated the effect of the most widely prescribed statin, atorvastatin, on the characteristics and properties of bone-marrow-derived mesenchymal stem cells (BM-MSCs) cultured in vitro. We found that atorvastatin did not decrease the viability of BM-MSCs, nor did it change the expression of MSC cell surface markers. Atorvastatin upregulated the mRNA expression levels of VEGF-A and HGF, whereas the mRNA expression level of IGF-1 was decreased. In addition, the PI3K/AKT signaling pathway was modulated by atorvastatin as indicated by the high mRNA expression levels of PI3K and AKT. Moreover, our data revealed the upregulation of mTOR mRNA levels; however, no change was observed in the BAX and BCL-2 transcripts. We propose that atorvastatin benefits BM-MSC treatment due to its ability to upregulate angiogenesis-related genes expression and transcripts of the PI3K/AKT/mTOR pathway.

Wound Healing Potential of Couroupita guianensis Aubl. Fruit Pulp Investigated on Excision Wound Model.

Wound care management aims at stimulating and improving healing process without scar formation. Although various plants have been reported to possess wound healing properties in tribal and folklore medicines, there is a lack of scientific data to validate the claim. In this aspect, it becomes inevitable to prove the efficacy of naturally derived products at pharmacological levels. Couroupita guianensis as a whole plant has been reported to exhibit wound healing activity. The leaves and fruit of this plant have been utilized in folkloric medicine to cure skin diseases and infections for many years. However, to the best of our knowledge, no scientific studies have been conducted to verify the wound healing properties of C. guianensis fruit pulp. Therefore, the present study seeks to investigate the wound healing potential of C. guianensis fruit pulp using an excision wound model in Wistar albino male rats. This study indicated that the ointment prepared from crude ethanolic extract of C. guianensis fruit pulp facilitated wound contraction that were evidenced by a greater reduction in the wound area and epithelialization period and increased hydroxyproline content. The experimental groups treated with low and mid dose of C. guianensis ethanol extract (CGEE) ointments had shown a wound closure of 80.27% and 89.11% respectively within 15days, which is comparable to the standard betadine ointment which showed 91.44% healing in the treated groups. Further, the extract influenced the expression of genes VEGF and TGF-β on post wounding days that clearly explained the strong correlation between these genes and wound healing in the experimental rats. The animals treated with 10% CGEE ointment showed a significant upregulation of both VEGF and TGF-β as compared with other test and standard groups. These findings provide credence to the conventional application of this plant in the healing of wounds and other dermatological conditions, and may represent a therapeutic strategy for the treatment of wounds.

The inhibitory effect of 6-gingerol and cisplatin on ovarian cancer and antitumor activity: In silico, in vitro, and in vivo.

Epithelial ovarian cancer is very common in women and causes hundreds of deaths per year worldwide. Chemotherapy drugs including cisplatin have adverse effects on patients' health. Complementary treatments and the use of herbal medicines can help improve the performance of medicine. 6-Gingerol is the major pharmacologically active component of ginger. In this study, we compared the effects of 6-gingerol, cisplatin, and their combination in apoptotic and angiogenetic activities in silico, in test tubes, and in in vivo assays against two ovarian cancer cell lines: OVCAR-3 and human umbilical vein endothelial cells (HUVECs). The drug-treated cell lines were evaluated for their cytotoxicity, cell cycle, and apoptotic and angiogenetic gene expression changes. The proportion of apoptosis treated by 6-gingerol coupled with cisplatin was significantly high. In the evaluation of the cell cycle, the combination therapy also showed a significant promotion of a higher extent of the S sequence. The expression of p53 level, Caspase-8, Bax, and Apaf1 genes was amplified again with combination therapy. Conversely, in both cell lines, the cumulative drug concentrations reduced the expression of VEGF, FLT1, KDR, and Bcl-2 genes. Similarly, in the control group, combination treatment significantly decreased the expression of VEGF, FLT1, KDR, and Bcl-2 genes in comparison to cisplatin alone. The findings of the present study demonstrated that the cisplatin and 6-gingerol combination is more effective in inducing apoptosis and suppressing the angiogenesis of ovarian cancer cells than using each drug alone.

Preeclampsia association of placental nucleotide variations in eNOS, VEGFA, and FLT-1 genes in Latin American pregnant women

IntroductionPreeclampsia is a leading cause of maternal and fetal morbidity in low- and middle-income countries, including those in Latin America. Placental vascular alterations are crucial in the pathophysiology of preeclampsia and few studies have evaluated nucleotide variations on genes associated with vascular regulation in the human placenta. This study aimed to evaluate whether placental nucleotide variations on eNOS, VEGFA, and FLT-1 genes are more frequently associated with preeclampsia in the Latin American population. MethodsThis case-control study included placental tissue from 88 controls and 82 cases that were genotyped through Taqman probes for eNOS, VEGFA, and FLT-1 genes. The intergroup comparisons were analyzed with the Mann-Whitney U test. Genotype and allele frequencies were compared by the X2 test. The association between the nucleotide variants with preeclampsia was evaluated through logistic regression analysis. ResultsA significant association was observed for VEGFA SNV rs2010963 (OR 1.95; CI 95% 1.13–3.37), after adjusting for population substructure. The allele combination T, G, G, C, C, C (rs2070744, rs1799983, rs2010963, rs3025039, rs699947 and rs4769613 respectively), showed a negative association with preeclampsia (OR 0.08; CI 95% 0.01–0.93). results. DiscussionPlacental SNV rs2010963 in the VEGFA gene was a risk factor for preeclampsia, while the allele combination T, G, G, C, C, C may represent potential protective factors for preeclampsia within Latin American women.

Restricted feeding regimens improve white striping associated muscular defects in broiler chickens.

The current study investigated the effects of intermittent feeding (IF) and fasting strategies at different times post-hatch on muscle growth and white striping (WS) breast development. In the first trial, 32 one-day-old Abor Acre broilers were fed ad libitum (AL) for 3d post-hatch and then randomly allotted into 4 feeding strategies including AL, 1h-IF group (1h IF, 4 times feeding/d, 1h each time), 1.5h-IF (1.5h IF, 4 times feeding/d, 1.5h each time), and fasting (1d acute fasting, 6d free access to feed) groups and fed for 7d. Although angiogenic genes including VEGFA, VEGFR1, and VEGFR2, and myogenic genes including MYOG and MYOD were upregulated (P<0.05), the breast muscle satellite cell (SC) number and PAX7, MYF5 expression were decreased by the IF strategies (P<0.05). One-day fasting at 6d of age also upregulated angiogenic genes and MYOD expression (P<0.05), downregulated MYF5 expression (P<0.05), but did not change SC number (P>0.05). In the second trial, 384 one-day-old birds were fed AL for 1wk and then randomly allotted to the above 4 feeding strategies starting at 8d of age until 42d of age. Similarly, IF and fasting strategies upregulated the expression of angiogenic and myogenic genes (P<0.05). Both 1h-IF and 1.5h-IF increased breast muscle SC number (P<0.05). At slaughter, breast muscle fiber diameter of 1.5h-IF was smaller but the SC number was larger than that of the birds fed AL (P<0.05). The IF and fasting strategies prevented WS development, and reduced breast WS scores and triglyceride content (P<0.05) without changing the body weight (P>0.05). Fasting and 1h-IF reduced the expression of adipogenic genes ZNF423 and PDGFRα (P<0.05). Moreover, IF and fasting strategies reduced fibrosis in breast muscle and reduced skeletal muscle-specific E3 ubiquitin ligases (TRIM63 and MAFBX) (P<0.05). Fasting significantly reduced CASPASE-3 in breast muscle (P<0.05). In conclusion, IF starting in the first week decreases SC number. Compared to AL, IF or fasting promotes muscular angiogenesis, increases SC number, prevents muscle degeneration, and prevents the development of WS without impairing the growth performance of broiler chickens.

Role of p65 NF-κB, caspase-3 activities and VEGF gene polymorphisms on the development of preterm labor in women with placental dysfunction

The objective: to study the activity of the p65 subunit of the nuclear factor – NF-κB and effector caspase-3, as well as the presence of a single nucleotide polymorphism of the VEGF gene (936C/T) in pregnant women with placental disorders and various clinical types of the course of premature labor (with premature rupture of the membranes and without it).Materials and methods. At the clinical base of the Department of Obstetrics and Gynecology No. 1 of O. O. Bogomolets National Medical University, which is located in the non-commercial enterprise “Perinatal Center of Kyiv”, during 2019–2022, a case-control study was conducted with the involvement of 90 pregnant women: 60 women of the main group with placental disorders and the development of spontaneous premature labor in the period of 24–34 weeks of gestation (I group – 30 women with premature labor and premature rupture of membranes, II group – 30 women with premature rupture of the membranes) and 30 women of control group (CG) with term normal delivery and the physiological course of the pregnancy.In pregnant women, the number of total and activated fractions of the p65 subunit of nuclear factor NF-κB and caspase-3 was determined by enzyme-linked immunosorbent assay (ELISA) in placenta lysates with further calculation of their activity based on these data, as well as the presence of a single nucleotide polymorphism of the VEGF gene (936C/T) using the polymerase chain reaction.Results. In women with placental disorders and preterm birth a significantly higher values of fraction content and activity of p65 NF-κB and caspase-3 were found, with some peculiarities within the groups compared to the control group. Women of the I group were characterized by a higher activity of p65-subunit of nuclear factor (I group — 61.6 % with 95 % CI 59.7–64.2; II group — 33.8 pg/ml with 95 % CI 31.2-35.2; CG — 27.3 pg/ml, 95 % CI 26.4–28.6; p&lt;0.05). Pregnant women of the II group had higher values of caspase-3 activity (II group — 59.2 % with 95 % CI 57.4–59.8, I group — 39.5 % with 95 % CI 38.5–40.5, CG — 31.2 %, 95 % CI 30.4–31.9; p&lt;0.01). It was established that the presence of the T allele at position 936 of VEGF gene polymorphism is a risk factor for the development of placental disorders with the development of premature labor, the rate of this allele in the main group was 11.7 % versus 1.7 % – in the control cohort (p&lt;0.05).Conclusions. Pregnant women with placental disorders and the development of premature birth are characterized by an increased activity of the p65 subunit of nuclear factor κB and caspase-3 compared to the control group: in women without premature rupture of the fetal membranes, a significant 2.2-fold increase in p65 NF-κB activity was found, and caspase-3 activity – by 1.3 times; in the group of pregnant women with preterm premature rupture of membranes the level of caspase-3 activity exceeded the control group by 1.9 times, and the level of p65-subunit of nuclear factor activity — by 1.2 times (p&lt;0.01 for all groups).936(C/T) single nucleotide polymorphism of the vascular endothelial growth factor gene was determined significantly more often in women with placental disorders and preterm birth (11.7 % in the study group versus 1.7 % in CG, p&lt;0.05), the T-allele on the VEGF gene carriers may be associated with the development of these pregnancy complications. However, a study on a larger sample of women is needed to obtain definitive results.

Controlling the Stem Cell Environment Via Conducting Polymer Hydrogels to Enhance Therapeutic Potential

AbstractStem cells are a promising treatment option for various neurological diseases such as stroke, spinal cord injury, and other neurodegenerative disorders. However, the ideal environment to optimize the therapeutic potential of the cells remains poorly understood. Stem cells in the native environment are influenced by a combination of mechanical, chemical, and electrical cues for proliferation and differentiation. Because of their controllable properties, conductive hydrogels are promising biomaterials to interact with stem cells. Herein, this work develops an interpenetrating conducting polymer hydrogel with tunable mechanical properties. The hydrogel serves as a platform to provide mechanical and electrical cues for interactions with mesenchymal stem cells (MSCs). This work optimizes the formulation of the hydrogel for maximum viability of MSCs and relatively higher cytoskeletal protein expression. The viability of cells is not affected due to electrical stimulation (ES). Further, ES alters the trophic factor secretion of MSCs, with significant increase in VEGF pathway genes—VEGFA and HSPB1. In addition, substrate stiffness of the hydrogel enhances the VEGFB secretion compared to control. Hence, the conducting polymer hydrogel system creates a tunable physical and electrical niche to enhance the therapeutic potential of stem cells for neurological injuries.

Chemical hazard of robotic hull in-water cleaning discharge on coastal embryonic fish

In-water cleaning (IWC) involves the removal of biofilms and foulants from the hull of a ship using brush or water jet. During IWC, several factors associated with the harmful chemical contaminants release to the marine environment, which can create “hotspots” of chemical contamination in coastal areas. To elucidate the potential toxic effects of IWC discharge, we investigated developmental toxicity in embryonic flounder, which are sensitive life stage to chemical exposure. Zinc and copper were the dominant metals, while zinc pyrithione was the most abundant biocide associated with IWC discharge in two remotely operated IWC. Discharge from IWC carried by both remotely operated vehicles (ROVs) produced developmental malformations including pericardial edema, spinal curvature, and tail-fin defects. In an analyses of differential gene expression profiles (fold-change of genes with a cutoff < 0.05) as assessed by high-throughput RNA sequencing, genes associated with muscle development were commonly and significantly changed. The gene ontology (GO) of embryos exposed to IWC discharge from ROV A activities highly enriched muscle and heart development, while cell signaling and transport were evident in embryos exposed to IWC discharge of ROV B. We analyzed the gene network by significant GO terms. In the network, TTN, MYOM1, CASP3, and CDH2 genes appeared to be key regulators of the toxic effects on muscle development. In embryos exposed to ROV B discharge, HSPG2, VEGFA, and TNF genes related to the nervous system pathway were affected. These results shed light on the potential impacts of muscle and nervous system development in non-target coastal organisms exposed to contaminants found in IWC discharge.

Plasmid containing VEGF-165 and ANG-1 dual genes packaged with fibroin-modified PEI to promote the regeneration of vascular network and dermal tissue

Reducing the cytotoxicity of cationic polymers is the major issue to their use as a gene delivery carrier. In this study, plasmids containing encoding vascular endothelial cell growth factor 165 and angiopoietin-1 were packaged with the conjugates of cationic fibroin (CSF) and polyethylenimine (PEI), instead of packaging pDNA with PEI alone, to prepare nanocomplexes (CSF+PEI)/pDNA. The complexes were loaded into a silk fibroin scaffold to enhance its function to induce microvascular network generation and dermal tissue regeneration. The results of transfecting EA.hy926 cells with the complexes in vitro showed that (CSF+PEI)/pDNA had a stronger transfection ability than PEI/pDNA. Importantly, compared with PEI as the gene carrier alone, the cell viability was significantly increased and the cytotoxicity was effectively reduced after the conjugate of CSF and PEI was used as the gene carrier. The results of angiogenesis in chick embryo chorioallantoic membranes showed that compared with scaffolds loaded with PEI/pDNA, the neovascularization ratio in scaffolds loaded with (CSF+PEI)/pDNA was significantly increased. In vivo experimental results of scaffolds implantation for full-thickness skin defects in SD rats showed that, compared with loading PEI/pDNA complex, loading (CSF+PEI)/pDNA complex in the scaffold more effectively promoted the formation of vascular network in the scaffold and accelerated the regeneration of dermal tissue. The gene delivery system established in this study has application potential not only in the regeneration of vascular-containing tissues, but also in tumor gene therapy.

Chondromodulin is necessary for cartilage callus distraction in mice.

Chondromodulin (Cnmd) is a glycoprotein known to stimulate chondrocyte growth. We examined in this study the expression and functional role of Cnmd during distraction osteogenesis that is modulated by mechanical forces. The right tibiae of the mice were separated by osteotomy and subjected to slow progressive distraction using an external fixator. In situ hybridization and immunohistochemical analyses of the lengthened segment revealed that Cnmd mRNA and its protein in wild-type mice were localized in the cartilage callus, which was initially generated in the lag phase and was lengthened gradually during the distraction phase. In Cnmd null (Cnmd-/-) mice, less cartilage callus was observed, and the distraction gap was filled by fibrous tissues. Additionally, radiological and histological investigations demonstrated delayed bone consolidation and remodeling of the lengthened segment in Cnmd-/- mice. Eventually, Cnmd deficiency caused a one-week delay in the peak expression of VEGF, MMP2, and MMP9 genes and the subsequent angiogenesis and osteoclastogenesis. We conclude that Cnmd is necessary for cartilage callus distraction.

Characterization of the Dynamic Gastrointestinal Digests of the Preserved Eggs and Their Effect and Mechanism on HepG2 Cells.

Preserved eggs, an alkaline-fermented food, have been widely searched for their anti-inflammatory activity. Their digestive characteristics in the human gastrointestinal tract and anti-cancer mechanism have not been well explained. In this study, we investigated the digestive characteristics and anti-tumor mechanisms of preserved eggs using an in vitro dynamic human gastrointestinal-IV (DHGI-IV) model. During digestion, the sample pH dynamically changed from 7.01 to 8.39. The samples were largely emptied in the stomach with a lag time of 45 min after 2 h. Protein and fat were significantly hydrolyzed with 90% and 87% digestibility, respectively. Moreover, preserved eggs digests (PED) significantly increased the free radical scavenging activity of ABTS, DPPH, FRAP and hydroxyl groups by 15, 14, 10 and 8 times more than the control group, respectively. PED significantly inhibited the growth, cloning and migration of HepG2 cells at concentrations of 250-1000 μg/mL. Meanwhile, it induced apoptosis by up/down-regulating the expression of the pro-apoptotic factor Bak and the anti-apoptotic gene Bcl-2 in the mitochondrial pathway. PED (1000 μg/mL) treatment resulted in 55% higher ROS production than the control, which also led to apoptosis. Furthermore, PED down-regulated the expression of the pro-angiogenic genes HIF-1α and VEGF. These findings provided a reliable scientific reference for the study of the anti-tumor activity of preserved eggs.

Effect of VEGF on neurological impairment and prognosis of acute cerebral infarction patients: A retrospective case-control study.

Due to the complex pathological mechanism of acute cerebral infarction, the role of vascular endothelial growth factor (VEGF) on the disease is not clear. Therefore, a retrospective case-control study was performed to explore the effect of VEGF on neurological impairment and prognosis of acute cerebral infarction patients. A total of 100 patients with acute cerebral infarction admitted to our hospital from April 2021 to April 2022 were selected. Blood samples from all patients would be routinely collected to detect the expression of serum VEGF. Pearson chi-square, Spearman correlation and univariate Logistic regression were used to analyze the clinical data to explore the relationship between VEGF expression and basic information, stroke degree, quality of life, and prognosis of patients. To determine whether VEGF can provide relevant basis for the early prevention and prognostic treatment of acute cerebral infarction. And multivariate logistic regression was used to calculate the odds ratio between each variable and VEGF expression. Pearson chi-square test and Spearman correlation coefficient showed that sex, degree of stroke, limb convulsions, loss of consciousness, hemiplegia, aphasia, mental functioning score, overall quality of life score, and short-term prognosis were significantly correlated with VEGF expression in 100 patients. Univariate logistic regression was used to describe the ORs and 95% confidence interval of subjects at the univariate level, and the degree of stroke (OR = 83.333, P < 0.001), tic of limbs (OR = 26.316, P < 0.001), loss of consciousness (OR = 23.256, P < 0.001), hemiplegia (OR = 62.500, P < 0.001), aphasia (OR = 76.923, P < 0.001), mental functioning score (OR = 7.937, P < 0.001), overall quality of life score (OR = 5.464, P < 0.001), short-term prognosis (OR = 37.037, P < 0.001) was significantly correlated with the high expression of VEGF. The level of serum VEGF was positively correlated with neurological impairment degree and prognosis in patients with acute cerebral infarction, the more severe the degree of stroke and the worse the prognosis.

Denuded Descemet's membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture.

Recent clinical studies suggest that retinal pigment epithelial (RPE) cell replacement therapy may preserve vision in retinal degenerative diseases. Scaffold-based methods are being tested in ongoing clinical trials for delivering pluripotent-derived RPE cells to the back of the eye. The aim of this study was to investigate human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells survival and behaviour on a decellularized Descemet's Membrane (DM), which may be of clinical relevance in retinal transplantation. DMs were isolated from human donor corneas and treated with thermolysin. The DM surface topology and the efficiency of the denudation method were evaluated by atomic force microscope, scanning electron microscopy and histology. hESC-RPE cells were seeded onto the endothelial-side surface of decellularized DM in order to determine the potential of the membrane to support hESC-RPE cell culture, alongside maintaining their viability. Integrity of the hESC-RPE monolayer was assessed by measuring transepithelial resistance. RPE-specific gene expression and growth factors secretion were assessed to confirm maturation and functionality of the cells over the new substrate. Thermolysin treatment did not affect the integrity of the tissue, thus ensuring a reliable method to standardize the preparation of decellularized DM. 24 hours post-seeding, hESC-RPE cell attachment and initial proliferation rate over the denuded DM were higher than hESC-RPE cells cultured on tissue culture inserts. On the new matrix, hESC-RPE cells succeeded in forming an intact monolayer with mature tight junctions. The resulting cell culture showed characteristic RPE cell morphology and proper protein localization. Gene expression analysis and VEGF secretion demonstrate DM provides supportive scaffolding and inductive properties to enhance hESC-RPE cells maturation. Decellularized DM was shown to be capable of sustaining hESC-RPE cells culture, thus confirming to be potentially a suitable candidate for retinal cell therapy.

Organization of self-advantageous niche by neural stem/progenitor cells during development via autocrine VEGF-A under hypoxia

BackgroundTissue stem cells are confined within a special microenvironment called niche. Stem cells in such a niche are supplied with nutrients and contacted by other cells to maintain their characters and also to keep or expand their population size. Besides, oxygen concentration is a key factor for stem cell niche. Adult neural stem/progenitor cells (NSPCs) are known to reside in a hypoxic niche. Oxygen concentration levels are lower in fetal organs including brain than maternal organs. However, how fetal NSPCs adapt to the hypoxic environment during brain development, particularly before pial and periventricular vessels start to invade the telencephalon, has not fully been elucidated.MethodsNSPCs were prepared from cerebral cortices of embryonic day (E) 11.5 or E14.5 mouse embryos and were enriched by 4-day incubation with FGF2. To evaluate NSPC numbers, neurosphere formation assay was performed. Sparsely plated NSPCs were cultured to form neurospheres under the hypoxic (1% O2) or normoxic condition. VEGF-A secreted from NSPCs in the culture medium was measured by ELISA. VEGF-A expression and Hif-1a in the developing brain was investigated by in situ hybridization and immunohistochemistry.ResultsHere we show that neurosphere formation of embryonic NSPCs is dramatically increased under hypoxia compared to normoxia. Vegf-A gene expression and its protein secretion were both up-regulated in the NSPCs under hypoxia. Either recombinant VEGF-A or conditioned medium of the hypoxic NSPC culture enhanced the neurosphere forming ability of normoxic NSPCs, which was attenuated by a VEGF-A signaling inhibitor. Furthermore, in the developing brain, VEGF-A was strongly expressed in the VZ where NSPCs are confined.ConclusionsWe show that NSPCs secret VEGF-A in an autocrine fashion to efficiently maintain themselves under hypoxic developmental environment. Our results suggest that NSPCs have adaptive potential to respond to hypoxia to organize self-advantageous niche involving VEGF-A when the vascular system is immature.

Integrated network pharmacology and in-silico approaches to decipher the pharmacological mechanism of Selaginella tamariscina in the treatment of non-small cell lung cancer

Background and purposeNon-small cell lung cancer (NSCLC) is a major pathological type of lung cancer and accounts for more than 80% of all cases. In healthcare management, it is challenging to understand the mechanism of NSCLC due to diverse spectra and the limited number of reported data. Selaginella tamariscina is an evergreen perennial plant, hermaphrodite, and used to treat numerous diseases, including NSCLC. In vitro research revealed the therapeutic importance of S. tamariscina in contrast to NSCLC, but the molecular mechanism is still unclear. In the present study, a network pharmacology technique was employed to uncover the active ingredients, their potential targets, and signaling pathways in S. tamariscina for the treatment of NSCLC. MethodsPutative ingredients of S. tamariscina and significant genes of NSCLC were retrieved from the public database after screening. The overlapped targets among S. tamariscina related compounds and NSCLC were predicted using Venn plot. Following that, a compound-target-disease network was constructed using Cytoscape to decipher the mechanism of S. tamariscina for NSCLC. KEGG pathway and GO enrichment analysis were performed to investigate the molecular mechanisms and pathways related to S. tamariscina for NSCLC treatments. Lastly, molecular docking and molecular dynamic simulation analysis were performed to validate the interaction that exists between compounds and target proteins. ResultsThe findings of the current analysis explored the compound–target–pathway network and figured out that Hinokiflavone, Heveaflavone, Neocryptomerin, Isocryptomerin, Apigenin, Sotetsuflavone, and Cryptomerin B decisively contributed to the development of NSCLC by affecting AKT1, EGFR, VEGFA, and GCK3B genes. Later, molecular docking and simulation analysis was conducted to validate the successful activity of the active compounds against potential targets. Lastly, it is concluded that predicted multi-target compounds of S. tamariscina will help in improving the body's sensitivity to NSCLC by regulating the expression of AKT1, EGFR, VEGFA, and GCK3B, which may act as potential therapeutic targets of NSCLC. ConclusionIntegrated network pharmacology and docking analysis revealed that S. tamariscina exerted a promising preventive effect on NSCLC by acting on diabetes-associated signaling pathways. The current findings propose that AKT1, VEGFA, EGFR, and GSK3B genes are promising and viable therapeutic targets to reduce the incidence of NSCLC, thereby exerting potential therapeutic effects in NSCLC. This approach introduces a groundwork for further research on the protective mechanisms of S. tamariscina for NSCLC and applications of network pharmacology in drug discovery.

Ginsenoside Rg3 Reduces the Toxicity of Graphene Oxide Used for pH-Responsive Delivery of Doxorubicin to Liver and Breast Cancer Cells.

Doxorubicin (DOX) is extensively used in chemotherapy, but it has serious side effects and is inefficient against some cancers, e.g., hepatocarcinoma. To ameliorate the delivery of DOX and reduce its side effects, we designed a pH-responsive delivery system based on graphene oxide (GO) that is capable of a targeted drug release in the acidic tumor microenvironment. GO itself disrupted glutathione biosynthesis and induced reactive oxygen species (ROS) accumulation in human cells. It induced IL17-directed JAK-STAT signaling and VEGF gene expression, leading to increased cell proliferation as an unwanted effect. To counter this, GO was conjugated with the antioxidant, ginsenoside Rg3, prior to loading with DOX. The conjugation of Rg3 to GO significantly reduced the toxicity of the GO carrier by abolishing ROS production. Furthermore, treatment of cells with GO-Rg3 did not induce IL17-directed JAK-STAT signaling and VEGF gene expression-nor cell proliferation-suggesting GO-Rg3 as a promising drug carrier. The anticancer activity of GO-Rg3-DOX conjugates was investigated against Huh7 hepatocarcinoma and MDA-MB-231 breast cancer cells. GO-Rg3-DOX conjugates significantly reduced cancer cell viability, primarily via downregulation of transcription regulatory genes and upregulation of apoptosis genes. GO-Rg3 is an effective, biocompatible, and pH responsive DOX carrier with potential to improve chemotherapy-at least against liver and breast cancers.

Whole Exome Sequencing Study Suggests an Impact of FANCA, CDH1 and VEGFA Genes on Diffuse Gastric Cancer Development.

Gastric cancer (GC) is one of the most common cancer types in the world with a high mortality rate. Hereditary predisposition for GC is not fully elucidated so far. The aim of this study was identification of possible new candidate genes, associated with the increased risk of gastric cancer development. Whole exome sequencing (WES) was performed on 18 DNA samples from adenocarcinoma specimens and non-tumor-bearing healthy stomach tissue from the same patient. Three pathogenic variants were identified: c.1320+1G>A in the CDH1 gene and c.27_28insCCCAGCCCCAGCTACCA (p.Ala9fs) of the VEGFA gene were found only in the tumor tissue, whereas c.G1874C (p.Cys625Ser) in the FANCA gene was found in both the tumor and normal tissue. These changes were found only in patients with diffuse gastric cancer and were absent in the DNA of healthy donors.

Underlying mechanisms for physical activity-induced brain plasticity

Physical activity is an important component in daily life which become more important when comes to health-related complications. Physical activity regardless the type and intensity, can bring beneficial effect for general health and can improve skills in several aspects including learning tasks and response to stimuli along with other health benefits. By recruiting regular physical activity and exercise, which happened for athletes, some structural and functional changes may occur in some regions of brain including increase in network connection, increase the speed of message transformation, increase in gray matter, synaptic plasticity and brain plasticity. There are several suggested mechanisms for physical activity-induced brain plasticity including increase in gene expression, BDNF, IGF-1, VEGF and activation of cellular cascade. Although the beneficial effects of physical activity on brain flexibility is well known which is reviled in athletes, the most effective type of activity which can affect brain flexibility the most, is not clear yet.