Elevated extracellular calcium ions accelerate the proliferation and migration of HepG2 cells and decrease cisplatin sensitivity

Hepatoblastoma is the most frequent liver malignancy in children.HepG2 has been discovered as a hepatoblastoma-derived cell line and tends to form clumps in culture.Intriguingly,we observed that the addition of calcium ions reduced cell clumping and disassociated HepG2 cells.The calcium signal is in connection with a series of processes critical in the tumorigenesis.Here,we demonstrated that extracellular calcium ions induced morphological changes and enhanced the epithelial-mesenchymal transition in HepG2 cells.Mechanistically,calcium ions promoted HepG2 proliferation and migration by up-regulating the phosphorylation levels of focal adhesion kinase(FAK),protein kinase B,and p38 mitogen-activated protein kinase.The inhibitor of FAK or Ca2+/calmodulin-dependent kinaseⅡ(CaMKⅡ)reversed the Ca2+-induced effects on HepG2 cells,including cell proliferation and migration,epithelial-mesenchymal transition protein expression levels,and phosphorylation levels of FAK and protein kinase B.Moreover,calcium ions decreased HepG2 cells'sensitivity to cisplatin.Furthermore,we found that the expression levels of FAK and CaMKⅡwere increased in hepatoblastoma.The group with high expression levels of FAK and CaMKⅡexhibited significantly lower ImmunoScore as well as CD8+T and NK cells.The expression of CaMKⅡwas positively correlated with that of PDCD1 and LAG3.Correspondingly,the expression of FAK was negatively correlated with that of TNFSF9,TNFRSF4,and TNFRSF18.Collectively,extracellular calcium accelerates HepG2 cell proliferation and migration via FAK and CaMKⅡand enhances cisplatin resistance.FAK and CaMKⅡshape immune cell infiltration and responses in tumor microenvironments,thereby serving as potential targets for hepatoblastoma.

Effects of anti-CD4 antibody treatment on calcium ions influx in peanut-sensitized C3H/HeJ mice

The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibodies for 4 weeks.Stimulation with PPE increased the specific immunoglobulin E(IgE),cytokine,histamine,and mMcp-1 levels,upregulated decorin(Dcn)expression,induced Ca^(2+) inflow in the spleen,and augmented the expression of the transcription factors GATA-3 and Foxp3,which resulted in Th2 and Treg cell activation.Notably,the Ca^(2+) levels were positively correlated with the histamine,interleukin(IL)-4,IL-5,and IL-13 levels,and negatively correlated with IL-10 levels.However,administration of anti-CD4 antibodies markedly alleviated allergic symptoms,activated T cells,and reduced Ca^(2+) inflow,cytokine,histamine,mMcp-1,and the IgHG3,CXCLI2,MMP2 and FABP4 gene.Our results indicated that anti-CD4 antibodies can ameliorate PPE-induced allergy,which is probably related to the suppression of Ca^(2+) inflow,and inhibiting histamine,cytokine and IgHG3,CXCL12,MMP2,and FABP4,thus exerting a protective effect against PPEsensitized food allergy.

Influence of copper ions and calcium ions on adsorption of CMC on chlorite

The effects of copper ions and calcium ions on the depression of chlorite using CMC（carboxymethyl cellulose） as a depressant were studied through flotation tests,adsorption measurements,ζ potential tests and co-precipitation experiments.The results show that the electrostatic repulsion between the CMC molecules and the chlorite surfaces hinders the approach of the CMC to the chlorite while the presence of copper ions and calcium ions enhances the adsorption density of CMC.The action mechanisms of these two types of ions are different.Calcium ions can not adsorb onto the mineral surfaces,but they can interact with the CMC molecules,thus reducing the charge of the CMC and enhancing adsorption density.Copper ions can adsorb onto the mineral surfaces,which facilitates the CMC adsorption through acid/base interaction.The enhanced adsorption density is also attributed to the decreased electrostatic repulsion between the CMC and mineral surfaces as copper ions reduce the surface charge of both the mineral surfaces and the CMC molecules.

Effects of clay and calcium ions on coal flotation

The microflotation of three single minerals,mixed coal–kaolinite and mixed coal–montmorillonite were examined to study the effects of clay and calcium ions on coal flotation.The results show that the ash content of flotation concentrate increases by 3%in the presence of clay minerals,and the ash content would further increase by 3%in co-presence of clay minerals and high concentration of Ca2+.Scanning electron microscope(SEM)images and elemental spectrum analysis indicate that fne clay particles that coat on the coal surface,which is called slime coating,can affect the coal flotation.The slime coating would be induced much more easily in the presence of Ca2+.

Effects of MCI-186 on intracellular calcium ion concentration and membrane fluidity of hippocampal neurons in a rat model of Alzheimer's disease

The present study observed the effects of MCI-186 on the intracellular calcium ion concentration （[Ca2＋]i） and membrane fluidity of hippocampal neurons in Alzheimer＇s disease （AD） model rats to validate the neuroprotective effects of MCI-186. Compared with normal and sham-operated rats, the escape latency was obviously prolonged, spanning platform times were obviously reduced, [Ca2＋]i was increased, and microviscosity η value was increased in AD rats. MCI-186 at 0.5 mg/mL and 0.2 mg/mL obviously shortened escape latency, increased spanning platform times and decreased [Ca2＋]i and microviscosity η values in AD rats at 7 and 17 days after induction of the model. The differences in the above-mentioned indices between normal and MCI-186-treated rats were statistically significant. MCI-186 at 0.2 mg/mL yielded better curative effects than 0.5 mg/mL MCI-186. These findings suggest that MCI-186 improves learning and memory abilities in AD rats by decreasing [Ca2＋]i and increasing membrane fluidity of hippocampal neurons.

Adsorption of Potassium and Calcium Ions by Variable Charge Soils

Interactions of potassium and calcium ions with four typical variable charge soils in South China were examined by measuring pK-0.5pCa value with a potassium ion-selective electrode and a calcium ion-selective electrode,and pK value with a potassium ion-selective electrode.The results showed that adsorption of potassium and calcium ions increased with soil suspension pH,and the tendency of the pK-0.5pCa value changing with pH differed with respect to pH range and potassium to calcium ratio.Adsorption of equal amount of calcium and potassium ions led to release of an identical number of protons,suggesting similar adsorption characteristics of these two ions when adsorbed by variable charge soils.Compared with red soil,latosol and lateritic red soil had higher adsorption selectivities for calcium ion.The red soil had a greater affinity for potassium ion than that for calcium ion at low concentration,which seems to result from its possession of 2:1 type minerals,such as vermiculite and mica with a high affinity for potassium ion.The results indicated that adsorption of potassium and calcium ions by the variable charge soils was chiefly caused by the electrostatic attraction between the cations and the soil surfaces.Moreover,it was found that sulfate could affect the adsorption by changing soil surface properties and by forming ion-pair.

Negative regulation of gamma-aminobutyric acid type A receptor on free calcium ion levels following facial nerve injury

Previous studies have demonstrated that muscarinic, and nicotinic receptors increase free Ca2＋ levels in the facial nerve nucleus via various channels following facial nerve injury. However, intracellular Ca2＋ overload can trigger either necrotic or apoptotic cell death. Gamma-aminobutyric acid （GABA）, an important inhibitory neurotransmitter in the central nervous system, exists in the facial nerve nucleus. It is assumed that GABA negatively regulates free Ca2＋ levels in the facial nerve nucleus. The present study investigated GABA type A （GABAA） receptor expression in the facial nerve nucleus in a rat model of facial nerve injury using immunohistochemistry and laser confocal microscopy, as well as the regulatory effects of GABAA receptor on nicotinic receptor response following facial nerve injury. Subunits α1, α3, α5, β1, β2, δ, and γ3 of GABAA receptors were expressed in the facial nerve nucleus following facial nerve injury. In addition, GABAA receptor expression significantly inhibited the increase in nicotinic receptor-mediated free Ca2＋ levels in the facial nerve nucleus following facial nerve injury in a concentration-dependent fashion. These results suggest that GABAA receptors exhibit negative effects on nicotinic receptor responses following facial nerve injury.

Effects of gamma-aminobutyric acid receptors on muscarinic receptor-mediated free calcium ion levels in the facial nucleus following facial nerve injury

Muscarinic receptors and nicotine receptors can increase free calcium ion levels in the facial nucleus via different channels following facial nerve injury. In addition, γ-aminobutyric acid A （GABAA） receptors have been shown to negatively regulate free calcium ion levels in the facial nucleus by inhibiting nicotine receptors. The present study investigated the influence of GABAA, γ-aminobutyric acid B （GABAB） and C （GABAc） receptors on muscarinic receptors in rats with facial nerve injury by confocal laser microscopy. GABAA and GABAB receptors exhibited significant dose-dependent inhibitory effects on increased muscarinic receptor-mediated free calcium ion levels following facial nerve injury. Results showed that GABAA and GABAB receptors negatively regulate muscarinic receptor effects and interplay with cholinergic receptors to regulate free calcium ion levels for facial neural regeneration.

Langmuir-Blodgett Films and Calcium Ion Coordination of Biliverdin and Its Amphiphilic Derivatives

Monolayer formation and LB film fabrication of amphiphilic derivative of biliverdin 1, diododecyl biliverdinamide [B(CONHC12H25)(2), 2] at an air-water interface on pure water subphase and subphase containing calcium ion were investigated and compared with 1. The coordination in ordered molecular films is much different from that in bulk solution. The formation of ligand-calcium complex was confirmed by X-ray photoelectron spectroscopy.

Role of Calcium Ion in Apoptosis of MD Cancer Cells Induced by Arsenic Trioxide

In order to observe the role of calcium ion in apoptosis of MD cancer cells induced by arsenic trioxide, inhibition percentage was detected by MTT assay; morphology changes were examined by fluorescence microscope; apoptosis was examined by DNA Ladder; [Ca^2＋]i was investigated by spectrofluorimeter in vitro on MDCC-MSB 1 cells. The results showed that As2O3 inhibited the proliferation of MDCC-MSB1 cells in concentration dependent manner （P〈0.05 or P〈0.01）; typical apoptosis character was observed by fluorescence microscope; DNA Ladder was observed; the [Ca^2＋]i was elevated significantly after the treatment of As203 （P〈0.05 or P〈0.01） and showed a dose-dependent manner. It is concluded that the calcium may play an important role in apoptosis of MD cancer cells induced by arsenic trioxide.

Effects of Calcium Ions on Thermodynamic Properties of Mixed Bilirubin/Cholesterol Monolayers

The mixed monolayer behavior of bilirubin/cholesterol was studied through surface pressure-area （π-A） isotherms on aqueous solutions containing various concentrations of calcium ions. Based on the data of π-A isotherms, the mean area per molecule, collapse pressure, surface compressibility modulus, excess molecular areas, free energy of mixing, and excess free energy of mixing of the monolayers on different subphases were calculated. The results show an expansion in the structure of the mixed monolayer with Ca^2＋ in subphase, and non-ideal mixing of the components at the air/water interface is observed with positive deviation from the additivity rule in the excess molecular areas. The miscibility between the components is weakened with the increase of concentration of Ca^2＋ in subphase. The facts indicate the presence of coordination between Ca^2＋ and the two components. The mixed monolayer, in which the molar ratio of bilirubin to cholesterol is 3：2, is more stable from a thermodynamic point of view on pure water. But the stable 3：2 stoichiometry complex is destroyed with the increase of the concentration of Ca^2＋ in subphase. Otherwise, the mixed monolayers have more thermodynamic stability at lower surface pressure on Ca^2＋ subphase.

Luminometric Studies of Yeast Response to Complex Environmental Calcium Variations Demonstrate Sensing of External Calcium Ion Changes

The response of yeast to sharp environmental increases in calcium concentration has been extensively studied. However, systematic studies of the response under more general changes are still lacking. Only limited exploration of cellular responses has been conducted where calcium concentration is decreased. This article describes a set of luminometric experiments that monitor the cytosolic calcium concentration under changing external concentration conditions. As a decrease in external calcium concentrations requires the use of large sample volumes, the experiments require the use of equipment adapted for this purpose. We describe the modification of commercial luminometric equipment to make the exploration possible. We explore the yeast cellular behavior when an increase in external calcium concentration is followed by a decrease in external calcium concentration. We compare these results with those from the case of a double pulse of concentration increase. Results from the experiment show that the first, concentration increasing pulse produces the well-known sharp increase in cytosolic calcium followed by calcium sequestration to return to a cytosolic concentration near its initial condition. Surprisingly, the calcium decrease step shows similar results with a cytosolic increase followed by a return to lower levels. The results suggest the presence of a calcium sensing mechanism regulating calcium influx from external sources. This mechanism would produce channel opening as a response to any changes in external concentration, be it an enhancement or a depletion.

Bio-cementation for tidal erosion resistance improvement of foreshore slopes based on microbially induced magnesium and calcium precipitation

In most coastal and estuarine areas,tides easily cause surface erosion and even slope failure,resulting in severe land losses,deterioration of coastal infrastructure,and increased floods.The bio-cementation technique has been previously demonstrated to effectively improve the erosion resistance of slopes.Seawater contains magnesium ions(Mg^(2+))with a higher concentration than calcium ions(Ca^(2+));therefore,Mg^(2+)and Ca^(2+)were used together for bio-cementation in this study at various Mg^(2+)/Ca^(2+)ratios as the microbially induced magnesium and calcium precipitation(MIMCP)treatment.Slope angles,surface strengths,precipitation contents,major phases,and microscopic characteristics of precipitation were used to evaluate the treatment effects.Results showed that the MIMCP treatment markedly enhanced the erosion resistance of slopes.Decreased Mg^(2+)/Ca^(2+)ratios resulted in a smaller change in angles and fewer soil losses,especially the Mg^(2+)concentration below 0.2 M.The decreased Mg^(2+)/Ca^(2+)ratio achieved increased precipitation contents,which contributed to better erosion resistance and higher surface strengths.Additionally,the production of aragonite would benefit from elevated Mg^(2+)concentrations and a higher Ca^(2+)concentration led to more nesquehonite in magnesium precipitation crystals.The slopes with an initial angle of 53°had worse erosion resistance than the slopes with an initial angle of 35°,but the Mg^(2+)/Ca^(2+)ratios of 0.2:0.8,0.1:0.9,and 0:1.0 were effective for both slope stabilization and erosion mitigation to a great extent.The results are of great significance for the application of MIMCP to improve erosion resistance of foreshore slopes and the MIMCP technique has promising application potential in marine engineering.

Novel Role of Calcium-Sensitive Receptors in Chronic Hypoxia-Induced Proliferation of Pulmonary Vein Smooth Muscle Cells

Objective:Vascular remodeling due to chronic hypoxia(CH)occurs not only in the pulmonary arteries but also in the pulmonary veins.Pulmonary vascular remodeling arises from the proliferation of pulmonary vascular myocytes.However,the mechanism by which CH induces the proliferation of pulmonary vein smooth muscle cells(PVSMCs)is unknown.This study aimed to investigate the mechanism by which CH affects the proliferation of PVSMCs.Methods:PVSMCs were isolated from rat distal pulmonary veins and exposed to CH(4%O2,60h),and the expression of the calcium-sensitive receptor(CaSR)was detected by Western blotting and immunofluorescence.MTT assay was used to detect the proliferation viability of the cells,and the changes in the intracellular calcium concentration were detected by laser confocal scanning technique.Results:CaSR expression was present in rat distal PVSMCs,and CaSR protein expression was upregulated under hypoxia.The positive regulator spermine not only enhanced CH-induced CaSR upregulation but also enhanced CH-induced increase in cell viability and calcium ion concentration.The negative CaSR regulator NPS2143 not only attenuated CH-induced CaSR upregulation but also inhibited CH-induced cell viability and calcium ion concentration.Conclusion:CaSR-mediated hyperproliferation is a novel pathogenic mechanism for the development of proliferation in distal PVSMCs under CH conditions.

Flotation separation of spodumene and albite with activation of calcium ion hydrolysate components

It is difficult to separate spodumene and albite by flotation with calcium ion as activator.In this study,microflotation experiment,solution chemistry calculation,density functional theory(DFT)calculation and X-ray photoelectron spectroscopy(XPS)analysis were conducted to investigate the effect mechanism of c alcium hydrolysate on the flotation separation of spodumene and albite.Results show that the weak interaction intensity of Ca^(2+)with O atoms on the mineral surface makes it difficult to activate minerals effectively.The O atom in the hydroxyl group of Ca(OH)^(+)and Ca(OH)_(2)can be strongly bonded with Al atom on the mineral surface,which greatly improves the adsorption strength.After the activation of Ca(OH)^(+),the adsorption forms of collectors on the surface of spodumene and albite are quite different,and the adsorption intensity on the surface of spodumene is higher,which is conducive to the flotation separation of spodumene and albite.After the action of Ca(OH)_(2),the adsorption of the collector on the albite surface is stronger than that of spodumene,which is not conducive to the flotation separation of the two minerals.By adjusting the initial concentration of calcium ions and pulp pH,Ca(OH)^(+)is the main active component,which is the key to improve the separation effect of spodumene and albite.

Effect of calcium ions on the secondary structures of photosystemⅡ and its relations with photoinhibition

Calcium ions play an important role in the oxygen_evolving process of photosystem Ⅱ as demonstrated in many experiments. The changes of the secondary structures of PS Ⅱ induced by the depletion of Ca 2+ were reported. The results indicated that the removal of Ca 2+ led to the transition of α helix to turns and sheet structures. While Ca 2+ was re_added to the media, only the structures changed to turns could be recovered. The protein conformational changes of PS Ⅱ during the donor side photoinhibition induced by the depletion of Ca 2+ were also studied. This showed that the protein conformational changes differed between the control and Ca 2+ _depleted samples in a short period of illumination (within 10 min). However, the changes became similar when the illumination time was increased.

Theoretical Studies on Complexes of Calcium Ion with Amino Acids

The complexes formed by calcium ion and 12 common amino acids were investigated systematically in the gas phase at the level of MP2/6-311＋＋G（d,p）//MP2/6-3 1G（d,p）.The results show that the salt-bridge structure is the most preferred motif for Ca2＋ binding aliphatic amino acids without heteroatom in the side chain,while charge-solvated（CS） structure is the most preferred motif for Ca2＋ binding other amino acids except for glutamine and lysine.IR spectra of Gln-Ca2＋ and Asn-Ca2＋ complexes were calculated and compared well with the available experiments.From the study in aqueous solution,the bidentate salt-bridge structure was determined to be the most favorable for all the twenty kinds of amino acids to chelate Ca2＋ to both the oxygen atoms of the negatively carboxylate group in the backbone.

Identification of key pathways and gene expression in the activation of mast cells via calcium flux using bioinformatics analysis

Mast cells are the main effector cells in IgE-associated allergic disorders,and we have reported that mucosal mast cells(MMCs)play a more important role in the development of food allergy(FA).IgE with antigen or calcium ionophore stimulation can lead to the activation of MMCs via a calcium-dependent pathway.The purpose of the present study was to identify gene signatures with IgE/antigen(dinitrophenyl-bovine serum albumin,DNP-BSA)or calcium ionophore(A23187)on the activation of MMCs.Differentially expressed genes between the two types of samples were identified with microarray analysis.Gene ontology functional and pathway enrichment analyses of differentially expressed genes were performed using the database for annotation,visualization,and integrated discovery software.The results showed that IgE/antigen and A23187 could induce degranulation,increase vacuoles,and elevate the cytosolic calcium concentration in MMCs.Furthermore,GeneChip analysis showed that the same 134 mRNAs were altered with IgE/DNP-BSA and A23187,suggesting that DNP-BSA/IgE and A23187 affect the same signal pathway partly in degranulation.KEGG analysis showed that the data were enriched in NF-κB,TNF,MAPK,transcription factor activity,DNA binding,and nucleic acid binding,suggesting that activation of MMCs is a complex process.The results provide new insights on MMCs activation.

Droplet microfluidic chip for precise monitoring of dynamic solution changes

In this work,an automated microfluidic chip that uses negative pressure to sample and analyze solutions with high temporal resolution was developed.The chip has a T-shaped channel for mixing the sample with a fluorescent indicator,a flow-focusing channel for generating droplets in oil,and a long storage channel for incubating and detecting the droplets.By monitoring the fluorescence intensity of the droplets,the device could detect changes in solution accurately over time.The chip can generate droplets at frequencies of up to 42 Hz with a mixing ratio of 1:1 and a temporal resolution of 3–6 s.It had excellent linearity in detecting fluorescein solution in the concentration range 1–5μM.This droplet microfluidic chip provides several advantages over traditional methods,including high temporal resolution,stable droplet generation,and faster flow rates.This approach could be applied to monitoring calcium ions with a dynamic range from 102 to 107 nM and a detection limit of 10 nM.

T-type calcium channel expression in cultured human neuroblastoma cells

Human neuroblastoma cells （SH-SY5Y） have similar structures and functions as neural cells and have been frequently used for cell culture studies of neural cell functions.Previous studies have revealed L-and N-type calcium channels in SH-SY5Y cells.However,the distribution of the low-voltage activated calcium channel （namely called T-type calcium channel,including Cav3.1,Cav3.2,and Cav3.3） in SH-SY5Y cells remains poorly understood.The present study detected mRNA and protein expres-sion of the T-type calcium channel （Cav3.1,Cav3.2,and Cav3.3） in cultured SH-SY5Y cells using real-time polymerase chain reaction （PCR） and western blot analysis.Results revealed mRNA and protein expression from all three T-type calcium channel subtypes in SH-SY5Y cells.Moreover,Cav3.1 was the predominant T-type calcium channel subtype in SH-SY5Y cells.