Effect of Lanthanum(Ⅲ) on Cytosolic Free Calcium in Isolated Rabbit Mature Osteoclasts

The effect of lanthanum （ Ⅲ ） （La^3 ＋ ） on cytosolic free calcium （ [ Ca^2 ＋ ] i ） in isolated rabbit mature osteoclasts was studied with the employment of fluo-3/AM as an intracellular calcium-sensitive fluorescent probe by using a confocal laser scanning microscope. La^3＋ does not alter basal [Ca^2＋ ]i levels and cell spread area at the concentration of 1.00 × 10^- 8 mol· L ^- 1. However, La^3 ＋ at higher concentrations （ 1. 00 × 10^ - 5 and 1.00 × 10^- 7 mol· L^- 1 ） decreases [ Ca^2 ＋ ] i levels and cell spread area, and greater decreases are observed for the higher concentrations of La^3 ＋ . Since [Ca^2 ＋ ]i affects cytoskeleton and the adhesion properties of osteoclasts, our results seem to suggest that La^3 ＋ inhibit bone resorption by decreasing [Ca^2＋]i in rabbit mature osteoclasts.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Lysophosphatidic acid induced nuclear translocation of nuclear factor-κB in Panc-1 cells by mobilizing cytosolic free calcium

AIM: To clarify whether Lysophosphatidic acid (LPA) activates the nuclear translocation of nuclear factor-κB (NF-κB) in pancreatic cancer. METHODS: Panc-1, a human pancreatic cancer cell line, was used throughout the study. The expression of LPA receptors was confirmed by reverse-transcript polymerase chain reaction (RT-PCR). Cytosolic free calcium was measured by fluorescent calcium indicator fura-2, and the localization of NF-κB was visualized by immunofluorescent method with or without various agents, which effect cell signaling. RESULTS: Panc-1 expressed LPA receptors, LPA1, LPA2 and LPA3. LPA caused the elevation of cytosolic free calcium dose-dependently. LPA also caused the nuclear translocation of NF-κB. Cytosolic free calcium was attenuated by pertussis toxin (PTX) and U73122, an inhibitor of phospholipase C. The translocation of NF-κB was similarly attenuated by PTX and U73122, but phorbol ester, an activator of protein kinase C, alone did not translocate NF-κB. Furthermore, the translocation of NF-κB was completely blocked by Ca2+ chelator BAPTA-AM. Thapsigargin, an endoplasmic- reticulum Ca2+-ATPase pump inhibitor, also promoted the translocation of NF-κB. Staurosporine, a proteinkinase C inhibitor, attenuated translocation of NF-κB induced by LPA. CONCLUSION: These findings suggest that protein kinase C is activated endogenously in Panc-1, and protein kinase C is essential for activating NF-κB with cytosolic calcium and that LPA induces the nuclear translocation of NF-κB in Panc-1 by mobilizing cytosolic free calcium.

Effect of Interleukin-1β on the Elevation of Cytoplasmic Free Calcium of the Cultured Hippocampal Neurons Induced by L-Glutamate

To investigate the intracellular mechanism that interleukin 1β (IL 1β) facilitates epileptic seizure and neuronal damage, the effect of IL 1β alone or IL 1β plus glutamate (Glu) on the intracellular free calcium ([Ca 2+ ] i) of single cultured hippocampal neuron was examined by using EPC 9 light electricity measurement system. The results showed that IL 1β of different concentrations (5×10 3 U/L, 10×10 3 U/L, 20×10 3 U/L, 30×10 3 U/L, 50×10 3 U/L, 100×10 3 U/L) failed to affect the neuronal [Ca 2+ ] i, but IL 1β could facilitate the augmentation of neuronal [Ca 2+ ] i induced by Glu in a dose dependent pattern. MK 801 inhibited the effect of Glu on [Ca 2+ ] i, and also inhibited the effect of IL 1β on [Ca 2+ ] i induced by Glu, while verapamil did not influence the effect of Glu or IL 1β. It is concluded that IL 1β, as a neuromodulator, can facilitate the activation of NMDA receptor by Glu, induce the increase of intracellular calcium, which enhances the excitement of neuron.

Rapid Inhibition of the Glutamate-induced Increase of Intracellular Free Calcium by Magnesium in Rat Hippocampal Neurons

By using Fura-2/AM, the effects of magnesium (Mg 2+) on the glutamate-induced increase of intracellular free calcium ([Ca 2+]i) in the cultured hippocampal neurons and the features were investigated by integrated photoelectric detecting system. The experiments were designed to three groups (The drug was spit to the cells for 20 s): Group A receiving 1×10 —5 mol/L glutamate; Group B receiving 1×10 —5 mol/L glutamate and1×10 —5 mol/L Mg 2+ simultaneously; Group C receiving 1×10 —5 mol/L glutamate again after [Ca 2+]i in group B back to the baseline. The results showed that in group A, [Ca 2+]i was obviously increased. In group B, the changes in [Ca 2+]i and the peak value were significantly decreased. Moreover, the elevation of Phase 1 was slowed down and Phase 2 was shortened to some extent, and the plateau phase between them was relatively prolonged. In group C, calcium oscillation similar to that in group A occurred, but both the Phase 1 and Phase 2 were shortened and the △[Ca 2+]i was slightly decreased. It was suggested that Mg 2+ could quickly inhibit the rise of [Ca 2+]i induced by glutamate in the cultured hippocampal neurons in rats.

Both Hypoxic Endothelial Cell Conditioned Medium and Hypoxia Elevate Intracellular Free Calcium in Pulmonary Artery Smooth Muscle Cells

By using Ca2+ -sensitive fluorescent probe, Fura-2 , the effects of endothelial cell-conditioned medium and hypoxia on intracellular free calcium ( [Ca2+]i) in cultured pulmonary artery smooth muscle cell (PASMC) were studied. Normoxic porcine pulmonary artery endothelial cell-conditioned medium (NPAECCM) obviously elevated [Ca2+]i in PASMC,whereas the hypoxic porcine pulmonary artery endothelial cell conditioned medium (HPAECCM)significantly elevated [Ca2+]i in PASMC much more than NPAECCM. Both the effects of NPAECCM and HPAECCM were dependent on the cultured endothelial cell extracellular calcium concentrations, ranged from 1.8 mmol/L to 2. 4 mmol/L.Meanwhile, hypoxia directly increased, which was partially inhibited by verapamil,[Ca2+]i in PASMC through Ca2+ influx pathway.The data suggest that the augmented regulation of endothelial cell on PASMC via Ca2+ second messenger system and the hypoxia-induced Ca2+ influx into PASMC,particularly the former, may be components of mechanisms underlying hypoxic pulmonary vasoconstriction and chronic pulmonary hypertension.

Effects of tetrandrine on cytosolic free calcium concentration in corpus cavernosum smooth muscle cells of rabbits

Aim:To study the relaxation mechanisms of tetrandrine(Tet)on the corpus cavernosum smooth muscle.Methods: The corpus cavernosum smooth muscle cells from New Zealand white rabbits were cultured in vitro.[Ca^(2+)]_i was measured by Fluorescence Ion Digital Imaging System,using Fluo-2/AM as a Ca^(2+)-sensitive fluorescent indicator. Results:Tet(1,10 and 100 μmol/L)had no effect on the resting [Ca^(2+)]_i(P>0.05).In the presence of extracellular Ca^(2+)(2.5 mmol/L),Tet(1,10 and 100 μmol/L)inhibited [Ca^(2+)]_i elevation induced by high K^+ and phenylephrine(PE) in a concentration-dependent manner(P<0.05).In calcium free solution containing egtaic acid,Tet(1 and 10 μmol/L) had no inhibitory effects on [Ca^(2+)]_i elevation induced by PE(P>0.05).However,Tet(100μmol/L)inhibited [Ca^(2+)]_i elevation induced by PE(P<0.05).Conclusion:Tet inhibited the Ca^(2+)influx from the extracellular site via voltage- activated Ca^(2+)channel and α_1-adrenoceptor-operated Ca^(2+)channel.At a high concentration,Tet might inhibit the cytosolic calcium pool release in cultured corpus cavernosum smooth muscle cells.This inhibitory action on [Ca^(2+)]_i might be one of the relaxation mechanisms of Tet on the corpus cavernosum smooth muscle.(Asian J Androl 2006 Jul;8:405-409)

A Validated Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) Method to Estimate Free Calcium and Phosphorus in In Vitro Phosphate Binding Study of Eliphos Tablets

An ICP-OES method has been developed to estimate Calcium and Phosphorous in In vitro phosphate binding study of Eliphos Tablets. The method is selective and is capable of detecting calcium and phosphorous in the presence of other trace elements. The method has been validated using RF power of 1500 watts, plasma flow of 15L/min, Nebuliser flow of 0.8 L/min and plasma view at radial mode for calcium and axial mode for phosphorus. The wavelength was monitored for calcium and phosphorous at 317.933 nm and 213.677 nm respectively. The method has been validated in terms of specificity, precision, linearity, accuracy, limit of quantification and ruggedness. The In vitro binding studies were performed for Eliphos Tablets at eight dif- ferent phosphate concentrations by incubating at 37.0?C and analysis was performed using the validated method to estimate free calcium and phosphorus. The objective of the study is to provide an alternate In vitro method to estimate the binding capacity of calcium acetate tablets to avoid the expensive in-vivo bio clinical studies.

Effects of tetrandrine on the changes of free calcium level in neutrophils after smoke inhalation injury in the rabbit

The changes of the cytoplasmic free calcium level in the neutrophils after smoke in-halation injury were observed in rabbits and then the effects of tetrandrine,a calcium antago-nist,on the changes of free calcium level were studied.It was found that the number of neu-trophils increased significantly preceded by a transient decrease in the blood and also increasedin the bronehoalveolar lavage fluid after smoke inhalation.and the level of cytoplasmic free calci-um in the blood neutrophil increased likewise.Administration of tetrandrine resulted in a reduc-tion of the neutrophils number in the lungs and the free calcium level in the blood neutrophils toalleviate the pulmonary injury due to smoke inhalation.It is believed that there is a close rela-tionship between the activation of neutrophils and the pathophysiological changes of the lungs,and tetrandrine can exert its therapeutic effects on the injury by decreasing the free calcium levelin the neutrophils to modulate their functions.

EFFECTS OF GINSENOSIDE Rb1 AND Rg1 ON SYNAPTOSOMAL FREE CALCIUM LEVEL, ATPase AND CALMODULIN IN RAT HIPPOCAMPUS

Calcium homeostasis in synaptosomes is altered during aging. The intrasynaptomal free calcium concentration ([Ca2+]i) was determined in 3- and 24-month-old rats treated with or without Rbl and Rgl. As expected, the [Ca2-]i level increased with age. Treatment with Rbl and Rgl elicited an obvious decrease of [Ca2+]i content, especially in aged rates. In addition, Rbl and Rgl significantly stimulated the activity of Na+, K+-ATPase while Rbl inhibited the activity of Ca2+, Mg2--ATPase arid calmodulin. In view of the close relationship of [Ca2-]i level with aging, the changes of [Ca2+]i induced by Rbl and Rgl, as shown by our results, might provide an explanation of the mechanisms of their antiaging function.

High concentration of calcium ions in Golgi apparatus

The interphase NIH3T3 cells were vitally fluorescentstained with calcium indicator fluo-3 and Glogi probe C6NBD-ceramide, and then the single cells were examined by laser scanning confocal microscopy (LSCFM) for subcellular distributions of Ca2+ and the location of Golgi apparatus. In these cells, the intracellular Ca2+ were found to be highly concentrated in the Golgi apparatus. The changes of distribution of cytosolic high Ca2+ region and the Golgi apparatus coincided with the cell cycle phase.In calcium free medium, when the plasma membrane of the cells which had been loaded with fluo-3/AM were permeated by digitonin, the fluorescence of the Golgi region decreased far less than that of the cytosol. Our results indicated that the Glogi lumen retained significantly high concentration of free calcium.

Effects of 3, 4-Dihydroxyacetophenone on Cytosolic Calcium in Pulmonary Artery Endothelial and Smooth Muscle Cells during Acute Hypoxia

Summary: The effects of 3, 4-Dihydroxyacetophenone (3, 4-DHAP) on cytosolic free calcium [Ca~2+ ]_i in pulmonary artery endothelia (PAECs) and smooth muscle cells (PASMCs) during acute hypoxia were studied. Porcine pulmonary artery endothelial and smooth muscle cells (PASMCs) were cultured primarily, and they were divided into 4 groups: groups incubated under normoxia or hypoxia and those with or without treatment with 3, 4-DHAP. The [Ca~2+ ]_i of both PAECs and PASMCs was measured by determining the fluorescence of fura 2 AM on spetrofluorometer. Our results showed that hypoxia caused significant elevation of [Ca~2+ ]_i, in both PAECs and PASMCs, 3, 4-DHAP could attenuate the hypoxic elevation of [Ca~2+ ]_i only in PASMCs but not in PAECs. It is concluded that 3, 4-DHAP decreases the hypoxic elevation of [Ca~2+ ]_i in PASMCs. This might contribute to its inhibitory effect on hypoxic pulmonary vasoconstriction.

ATP Evokes Calcium Transients in Single Pulmonary Artery Endothelial Cell in Primary Cultures

Effects of ATF on cytosclic free calcium ([Ca2+]i) in single porcine pulmonary artery endothelia cell were studied.Using a dual-wavelength excitation on microflurometry.it was found that ATP evoked a rapid transient in[Ca2+]i which was then followed by a maintained elevation of[Ca2+]i.The removal of extracellular Ca2+ abolished the maintained plateform, but exerted no obvious effect on the initial-transient.These results suggest that ATP stimulates both calcium release from intracellular calcium pool(s)and calcium influx across the Plasma membrane from extracellular space.

STUDY ON THE PLATELET INTRACELLULAR FREE Ca^(2+) IN NORMAL PREGNANCY

This study measured platelet intracellular free calcium (PFCa 2+ ) of 1003 normal pregnancies in different trimester. The purpose of this study was to find the alteration of PFCa 2+ in normal pregnancy, to provide scientific basis for forecasting and preventing PIH (pregnancy induced hypertension). The results showed that the PFCa 2+ concentration was stable and slightly increased with the progression of gestational weeks.

钢渣尾渣中游离氧化钙水浸消解实验研究

随着冶金行业技术高速发展,钢铁产量逐年递增,钢渣作为其副产物,产量快速增长,钢渣中游离氧化钙(f-CaO)对环境的影响较大。为探究影响钢渣水浸过程中f-CaO消解的因素,设计了不同液固比、钢渣粒径、换水次数、搅拌时间等因素对钢渣中游离氧化钙消解影响的实验。实验结果表明,钢渣粒径加上适当的搅拌时间,对游离氧化钙消解效果影响最大,实验原渣中游离氧化钙的含量为7.17%,当钢渣粒径从1 mm减小至0.15 mm时,游离氧化钙含量从5.21%减少至4.92%,同时增加搅拌时间5 min,粒径为0.15 mm时,游离氧化钙的含量进一步降低至2.45%,此条件下从节能角度考虑较为合理。

混凝土中离散分布的游离氧化钙潜在危害的检测推断

常用于测试游离氧化钙(f-CaO)对混凝土潜在危害的方法包括薄片和芯样试件沸煮测试等,但由于试件体量较小,对离散分布在混凝土中的f-CaO潜在危害测试存在较大偶然性。以某项目实体混凝土墙、板为研究对象,通过沸煮实验,从外观、厚度、超声波声时、抗压强度多方面对实体混凝土中混入过烧石灰对混凝土质量影响进行测试推断。结果表明:采用沸煮前后外观、厚度、超声波声时、抗压强度多方面测试的结果更全面、准确,对相关事故的防控以及质量检测具有一定参考意义。

脱硫废液处理钢尾渣制备路面砖及其性能研究

采用弱酸性脱硫废液对钢尾渣处理,设计和制备了钢尾渣基路面砖,通过离子色谱法、XRD、FTIR、TG-DSC分析了钢尾渣的微观组成,研究了不同预处理方法对钢尾渣基路面砖抗压、抗折强度及体积安定性的影响。试验结果表明:脱硫废液的pH值由浸泡钢尾渣前的5.70增至浸泡钢尾渣后的7.36;脱硫废液处理后,钢尾渣中f-CaO的含量降低25.3%,且钢尾渣的体积膨胀抑制效果明显,体积安定性增强;所制备的钢尾渣基路面砖力学性能及沸煮安定性均能满足设计要求。

sFLC、血清钙离子在多发性骨髓瘤患者诊断及预后判断中的应用价值

目的:探讨血清游离轻链(sFLC)、血清钙离子在多发性骨髓瘤(MM)患者诊断及预后中的临床应用价值。方法:选取2018年1月至2022年1月在河南省人民医院治疗的MM患者40例作为观察组,同时选取健康志愿者40例作为对照组,比较两组患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子等差异性,同时分析观察组不同国际分期系统(ISS)、不同化疗疗效及不同预后患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子等差异性。结果:观察组sFLC-κ水平[(98.39±21.19)对(12.01±4.45)mg/L]、sFLC-λ水平[(210.20±45.54)对(14.10±5.11)mg/L]、低钙血症比例(65%对0)均明显高于对照组(P<0.05),而sFLC-κ/λ比值[(0.44±0.10)对(0.87±0.12)]、血清钙离子[(1.98±0.46)对(2.42±0.40)mmol/L]明显低于对照组(P<0.05)。观察组ISS分期Ⅲ期患者sFLC-κ、sFLC-λ浓度、低钙血症比例和低钙血症病程均明显高于Ⅰ期和Ⅱ期患者(P<0.05),而sFLC-κ/λ、血清钙离子均明显低于Ⅰ期和Ⅱ期患者(P<0.05)。观察组有低钙血症患者sFLC-κ水平[(107.76±21.22)对(94.67±20.11)mg/L]、sFLC-λ水平[(245.54±41.12)对(205.54±50.22)mg/L]明显高于无低钙血症患者(P<0.05),而sFLC-κ/λ比值明显低于无低钙血症患者[(0.42±0.04)对(0.47±0.06);P<0.05]。化疗无效患者sFLC-κ水平[(107.29±20.14)对(91.11±18.92)mg/L]、sFLC-λ水平[(247.98±42.26)对(179.29±39.32)mg/L]明显高于化疗有效患者(P<0.05),而sFLC-κ/λ比值明显低于化疗有效患者[(0.43±0.10)对(0.50±0.09);P<0.05]。sFLC-κ、sFLC-λ、sFLC-κ/λ预测化疗无效的ROC曲线下面积分别为0.803、0.793和0.699,P<0.05。存活和死亡患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子、低钙血症比例和低钙血症病程比较差异无统计学意义(均P>0.05)。结论:sFLC、血清钙离子与MM患者ISS分期有关,sFLC水平在预测MM患者化疗疗效方面有一定应用价值,sFLC与血清钙离子在判断MM患者预后方面暂未发现有应用价值。