Caffeic acid hinders the proliferation and migration through inhibition of IL-6 mediated JAK-STAT-3 signaling axis in human prostate cancer

Background:Caffeic acid(CA)is considered a promising phytochemical that has inhibited numerous cancer cell proliferation.Therefore,it is gaining increasing attention due to its safe and pharmacological applications.In this study,we investigated the role of CA in inhibiting the Interleukin-6(IL-6)/Janus kinase(JAK)/Signal transducer and activator of transcription-3(STAT-3)mediated suppression of the proliferation signaling in human prostate cancer cells.Materials and Methods:The role of CA in proliferation and colony formation abilities was studied using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide(MTT)assay and colony formation assays.Tumour cell death and cell cycle arrest were identified usingflow cytometry techniques.CA treatment-associated protein expression of mitogen-activated protein kinase(MAPK)families,IL-6/JAK/STAT-3,proliferation,and apoptosis protein expressions in PC-3 and LNCaP cell lines were measured using Western blot investigation.Results:We have obtained that treatment with CA inhibits prostate cancer cells(PC-3 and LNCaP)proliferation and induces reactive oxygen species(ROS),cell cycle arrest,and apoptosis cell death in a concentration-dependent manner.Moreover,CA treatment alleviates the expression phosphorylated form of MAPK families,i.e.,extracellular signal-regulated kinase 1(ERK1),c-Jun N-terminal kinase(JNK),and p38 in PC-3 cells.IL-6 mediated JAK/STAT3 expressions regulate the proliferation and antiapoptosis that leads to prostate cancer metastasis and migration.Therefore,to mitigate the expression of IL-6/JAK/STAT-3 is considered an important target for the treatment of prostate cancer.In this study,we have observed that CA inhibits the expression of IL-6,JAK1,and phosphorylated STAT-3 in both PC-3 and LNCaP cells.Due to the inhibitory effect of IL-6/JAK/STAT-3,it resulted in decreased expression of cyclin-D1,cyclin-D2,and CDK1 in both PC-3 cells.In addition,CA induces apoptosis by enhancing the expression of Bax and caspase-3;and decreased expression of Bcl-2 in prostate cancer cells.Conclusions:Thus,CA might act as a therapeutical application against prostate cancer by targeting the IL-6/JAK/STAT3 signaling axis.

Calcium-Magnesium Ca/Mg Ratios and Their Agronomic Implications for the Optimization of Phosphate Fertilization in Rainfed Rice Farming on Acidic Ferralsol in the Forest Zone of Ivory Coast

This study is a contribution to improving rice productivity on acidic plateau soils of the tropical rainforest zone. It is based on taking into account the cationic balances of the soil in order to optimize the phosphorus (P) nutrition of rice on these acidic soils, where this nutrient constitutes a limiting factor for agricultural production. Three (3) pot trials were conducted in Adiopodoumé in the forested south of Côte d’Ivoire. The interactive effects of calcium carbonate (0, 25, 50 and 75 kg Ca ha−1) and magnesium sulfate (0, 25, 50 and 75 kg Mg ha−1) were evaluated on the response of NERICA 5 rice at doses 0, 25, 50 and 75 kg P ha−1 of natural phosphate from Togo, applied only once at the start of the experiment. Additional fertilizers of nitrogen (N) (100 kg N ha−1) and potassium (K) (50 kg KCl ha−1) were added to each of the tests in a split-plot device. The test results revealed a paddy production potential of approximately 3 to 5 t⋅ha−1 for NERICA 5 on an acidic soil, under the effect of the interaction of P, Ca and Mg. The quadratic response of rice yield to the doses of these fertilizers would be more dependent on their balance, itself influenced by Ca nutrition. For the sustainability and maintenance of rice production in agro-ecology studied, it was recommended doses of 38 kg Ca ha−1, 34 kg Mg ha−1 in a Ca/Mg ratio (1/1) with intakes of 41 kg P ha−1, overall in a ratio 1/1/1 (P/Ca/Mg) more favorable to the availability of free iron considered a guiding element of mineral nutrition. Thus, these promising results should be confirmed in a real environment for better management of the fertilization of rice cultivated on acidic plateau soils in Côte d’Ivoire.

酪氨酸激酶抑制剂对cyclopiazoni cacid引起的大鼠主动脉血管环收缩效应的影响

目的 探讨酪氨酸激酶在Ca2 +池操纵性Ca2 +内流中的作用。方法 记录大鼠胸主动脉环收缩反应。结果 ①不同剂量酪氨酸激酶抑制剂 genistein (1～ 10 0 μmol·L-1)和tyrphostin 2 5 (Tyr 2 5 ,1～ 30 μmol·L-1)均以浓度依赖性抑制cyclopiazonicacid (CPA)引起的平滑肌收缩平台期。Tyr 2 5的最大作用浓度为 10 μmol·L-1,抑制率为 42 %±11%。② 10 μmol·L-1Tyr 2 5和 1μmol·L-1nifedipine(Nif)对CPA引起电压依赖性Ca2 +通道 (VDCC)开放过程的抑制作用存在部分交叉。③ 1μmol·L-1Nif预处理阻断VDCC作用后 ,10 μmol·L-1Tyr 2 5只能部分阻断CPA引起的Ca2 +池操纵性Ca2 +通道 (SOCC)开放过程 ,再加入 6 0 μmol·L-1SK&F96 36 5可完全阻断SOCC的开放过程。结论 CPA引起平滑肌的收缩过程中 。

Effects of Calcium and Jasmonic Acid on CBF Expression in Spinach

[Objective] This experiment aimed to evaluate the effects of calcium and Jasmonic acid（JA） on the expression of CBF in spinach. [Methods] The seedlings of spinach were treated with low temperature （4 ℃）, JA or A23187, then used for detecting the expression of CBF by northern blotting. [Results] The results showed that the CBF expression was regulated by low temperature and JA positively. [Conclusions] Low temperature may increase the JA content of the cell firstly, then JA induced the increase of cytosolic calcium concentration （[Ca2＋]cyt）, and the JA induced Ca2＋ transmitted the low temperature signal through CaM or CaM related proteins, regulating the CBF expression.

IP_3 Sensitive Calcium Channel Involved in the Jasmonic Acid Induced Calcium Mobilization

[Objective] This study was to investigate the role of IP3 sensitive calcium channel in the JA-induced calcium mobilization pathway.[Method] Arabidopsis thaliana leaves were labeled by Fluorescent probe Fluo-3/AM under low temperature at 4 ℃ to measure the fluorescent intensity of intracellular Ca2＋ which was pretreated with heparin on jasmonic acid（JA）-induced.[Results] When A.thaliana leaf cells were pretreated with 10,50 or 100 ng/ml heparin,intercellular free Ca2＋ fluorescence intensity was reduced in comparison with negative control.Once the heparin-pretreated A.thaliana leaf cells were stimulated with 100 μmol/L JA,intercellular Ca2＋ fluorescence intensity increased gradually and tended to be stable at a degree equivalent with that in negative control.[Conclusion] The experiment showed that the pretreatment with heparin could inhibit the increase of the intracellular Ca2＋ concentration significantly which JA-induced in leaves of Arabidopsis thaliana.

富里酸和Ca^(2+)共存对嗜酸性氧化亚铁硫杆菌介导生成次生高铁矿物的影响

为揭示富里酸和Ca^(2+)共存对嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)氧化酸性矿山废水(AMD)中的Fe^(2+)和形成次生高铁矿物的影响,分析了pH、Fe^(2+)氧化率、铁沉淀率以及次生高铁矿物矿相、基团等相关指标。结果表明,Ca^(2+)确实具有提高嗜酸性氧化亚铁硫杆菌氧化Fe^(2+)的能力。低质量浓度(0.2 g/L)的富里酸对嗜酸性氧化亚铁硫杆菌活性的提高具有促进作用,高质量浓度(0.4 g/L)的富里酸具有抑制作用,而增加Ca^(2+)反过来能够减弱高浓度富里酸对嗜酸性氧化亚铁硫杆菌的抑制作用。对形成的次生高铁矿物进行X射线衍射(XRD)和傅立叶红外光谱(FTIR)分析,结果表明高浓度富里酸促进了另一次生高铁矿物草黄铁矾的生成。

氟碳铈矿除Ca、Si制备脱硝催化剂及其脱硝性能的研究

包头混合稀土精矿是氟碳铈矿(CeCO_(3)F)和独居石(CePO_(4))的共生矿物,结构复杂,难以分解,且Ca、Si杂质嵌布在矿物表面,影响了稀土矿物的脱硝活性,因此在使用天然矿物作为催化剂之前,有必要对其进行除杂预处理。采用酸浸固氟焙烧方案将氟碳铈矿制成脱硝催化剂,具体工艺为:第一步,HCl除Ca,HCl浓度为6mol/L,氟碳铈矿与NaCl(固氟剂)质量比为1∶5,混合并搅拌12h后于500℃下焙烧2h;第二步,HF除Si,HF浓度为13mol/L,除Ca后粉末与NaCl质量比为1∶5,混合并搅拌12h后于500°C下焙烧2h。氟碳铈矿除Ca、Si后制备的催化剂中Ca含量降至1.85%,Si含量降至0.17%,稀土元素含量相比原矿提高了约22%。氟碳铈矿除Ca、Si后,表面形成了发达的孔结构,比表面积增大,活性元素增多,且部分Ce的价态由+3价转变为+4价,还形成了晶格氧,这些组织结构的变化为催化剂脱硝过程提供了更多的氧化还原中心,使得催化剂脱硝性能显著提高。因此,用除杂后的氟碳铈矿作脱硝催化剂比直接用天然矿物具有更大的优势。

Effect of electroacupuncture on glial fibrillary acidic protein and nerve growth factor in the hippocampus of rats with hyperlipidemia and middle cerebral artery thrombus

Electroacupuncture(EA)has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia.However,there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects.In this study,EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed.Except for the normal group,rats in other groups were fed a high-fat diet throughout the whole experiment.Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks.Middle cerebral artery thrombus models were induced by pasting 50%FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group.EA1 group rats received EA at bilateral ST40(Fenglong)for 7 days before the thrombosis.Rats in the EA1 and EA2 groups received EA at GV20(Baihui)and bilateral ST40 for 14 days after model establishment.Neuronal health was assessed by hematoxylin-eosin staining in the brain.Hyperlipidemia was assessed by biochemical methods that measured total cholesterol,triglyceride,low-density lipoprotein and high-density lipoprotein in blood sera.Behavioral analysis was used to confirm the establishment of the model.Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region.The results demonstrated that,compared with the model group,blood lipid levels significantly decreased,glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups.The repair effect was superior in the EA1 group than in the EA2 group.These findings confirm that EA can reduce blood lipid,inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis.All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine,China(approval No.BUCM-3-2018022802-1002)on April 12,2018.

18β-glycyrrhetinic Acid-induced Apoptosis and Relation with Intracellular Ca^2＋ Release in Human Breast Carcinoma Cells

Objective:To study the effects of 18β-glycyrrhetinic acid (GA) on proliferation inhibition, apop totic induction, and the relationship between GA-induced apoptosis and intracellular Ca2+ concentration in human breast carcinoma (MCF-7) cells. Methods: After MCF-7 cells were treated with GA at the concentrations from 50 μmol/L to 250 μmol/L for 24 h, cell viability of proliferation was assessed by MTT assay. After the cells were treated with 100 μmol/L, 150 μmol/L, and 200 μmol/L GA for 24 h, the rates of cell apoptosis were examined by terminal deoxynucleotide transferase mediated dUTP nick-end-labeling method and flow cytometry with Annexin V/propidium iodide fluorescent stain. After the cells treated with 150 μmol/L GA for 24 h, intracellular Ca2+ concentration was measured by Fure-2 fluorescein load method. Results: After the cells were treated with GA at the concentrations from 100 μmol/L to 250 μmol/L, the rates of proliferative inhibition were increased significantly (P<0.05 and P<0.01) in a dose dependent fashion. IC50 of the proliferation inhibition was 234.33 μmol/L. Treated with 100 μmol/L, 150 μmol/L, and 200 μmol/L, the rates of cell apoptosis were increased significantly (P<0.01). Intracellular Ca2+ concentration after treatment with GA was higher evidently than that of control (P<0.05). Conclusion: 18β-glycyrrhetinic acid has the effects of the proliferation inhibition and the apoptotic induction on MCF-7 cells. The rise of intracellular Ca2+ level may be depended on apoptosis induced by GA in MCF-7 cells.

Targeted mutagenesis of amino acid transporter genes for rice quality improvement using the CRISPR/Cas9 system

High grain protein content(GPC) reduces rice eating and cooking quality(ECQ). We generated OsAAP6 and OsAAP10 knockout mutants in three high-yielding japonica varieties and one japonica line using the CRISPR/Cas9 system. Mutation efficiency varied with genetic background in the T_0 generation, and GPC in the T_1 generation decreased significantly,owing mainly to a reduction in glutelin content. Amylose content was down-regulated significantly in some Osaap6 and all Osaap10 mutants. The increased taste value of these mutants was supported by Rapid Visco Analysis(RVA) profiles, which showed higher peak viscosity and breakdown viscosity and lower setback viscosity than the wild type. There were no significant deficiencies in agronomic traits of the mutants. Targeted mutagenesis of OsAAP6 and OsAAP10, especially OsAAP10, using the CRISPR/Cas9 system can rapidly reduce GPC and improve ECQ of rice, providing a new strategy for the breeding cultivars with desired ECQ.

Chromoendoscopy of gastric adenoma using an acetic acid indigocarmine mixture

AIM: To investigate the usefulness of chromoendoscopy, using an acetic acid indigocarmine mixture (AIM), for gastric adenoma diagnosed by forceps biopsy.

Influence of Ca/P ratio on the catalytic performance of hydroxyapatite for decarboxylation of itaconic acid to methacrylic acid

Methacrylic acid,an important organic chemical,is commercially manufactured starting from fossil feedstock.The decarboxylation of itaconic acid derived for biomass is a green route to the synthesis of methacrylic acid.In view of the problems existing in the researches on this route such as use of noble metal catalyst,harsh reaction conditions and low desired-product yield,we prepared a series of hydroxyapatite catalysts with different Ca/P molar ratios and evaluated their catalytic performance.The results showed that the hydroxyapatite catalyst with a Ca/P molar ratio of 1.58 had the best catalytic activity.The highest yield of MAA up to 61.2%was achieved with basically complete conversion of itaconic acid under the suitable reaction conditions of 1 equivalent of NaOH,2 MPa of N_(2),250℃,and 2 h.On this basis,a reaction network for the decarboxylation of itaconic acid to methacrylic acid catalyzed by hydroxyapatite was established.With the aid of catalyst characterization using X-ray powder diffraction,NH3/CO2 temperature-programmed desorption,N_(2)physisorption,inductively coupled plasma optical emission spectrometry,and scanning electron microscopy,we found that the distribution of surface acid sites and basic sites,crystal growth orientation,texture properties and morphology of hydroxyapatite varied with the Ca/P molar ratio.Furthermore,the change of the crystal growth orientation and its influence on the surface acidity and alkalinity were clarified.

High-temperature stress suppresses allene oxide cyclase 2 and causes male sterility in cotton by disrupting jasmonic acid signaling

Cotton(Gossypium spp.) yield is reduced by stress. In this study, high temperature(HT) suppressed the expression of the jasmonic acid(JA) biosynthesis gene allene oxide cyclase 2(GhAOC2), reducing JA content and causing male sterility in the cotton HT-sensitive line H05. Anther sterility was reversed by exogenous application of methyl jasmonate(MeJA) to early buds. To elucidate the role of GhAOC2 in JA biosynthesis and identify its putative contribution to the anther response to HT, we created gene knockout cotton plants using the CRISPR/Cas9 system. Ghaoc2 mutant lines showed male-sterile flowers with reduced JA content in the anthers at the tetrad stage(TS), tapetum degradation stage(TDS), and anther dehiscence stage(ADS). Exogenous application of MeJA to early mutant buds(containing TS or TDS anthers) rescued the sterile pollen and indehiscent anther phenotypes, while ROS signals were reduced in ADS anthers. We propose that HT downregulates the expression of GhAOC2 in anthers, reducing JA biosynthesis and causing excessive ROS accumulation in anthers, leading to male sterility. These findings suggest exogenous JA application as a strategy for increasing male fertility in cotton under HT.

CRISPR-Cas系统在病原核酸检测中的研究进展

CRISPR-Cas系统作为原核生物获得性免疫系统,由簇状规则间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)和CRISPR相关蛋白(CRISPR-associated proteins,Cas)构成,因其识别和切割特定DNA或RNA序列,而成为分子诊断领域研究的热点。研究人员利用Cas蛋白(Cas12、Cas13、Cas14、Cas3等)结合信号放大和转化技术(荧光法、电位法、比色法、侧向流动技术等),开发了许多高灵敏度、高特异性、低成本的诊断平台,为病原核酸检测提供了新途径。本文介绍了CRISPR-Cas系统的生物学机制及分类,总结现有的基于Cas蛋白反式切割活性开发的病原核酸检测技术,描述其特点、功能和应用场景,并对该系统的未来应用前景进行展望,期望CRISPR-Cas系统成为包括核酸检测在内的多靶标的理想检测平台。

CRISPR/Cas-等温扩增技术在食源性病原菌检测中的研究进展

食源性病原菌引起的食品安全事件引起广泛关注,为保证食品安全,采取有效的检测手段至关重要。新兴的簇状规则间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)及其关联蛋白(CRISPR-associated protein,Cas)组成的CRISPR/Cas系统是广泛存在于细菌和古细菌中的一种获得性免疫系统,可高效、特异性识别并切割外源核酸,与传统技术相比在检测病原菌方面更为高效。本文基于CRISPR/Cas生物传感系统-等温扩增技术联用的相关研究进行综述,对CRISPR/Cas系统的分类、原理以及与等温扩增耦合后在食源性病原菌检测方面的研究进展进行概述,并对其在即时检测应用中的前景以及所面临的挑战进行深入探讨,以期为CRISPR/Cas生物传感系统的进一步发展提供参考。

Effects of Acetylsalicylic Acid and Calcium Chloride on Photosynthetic Apparatus and Reactive Oxygen-Scavenging Enzymes in Chrysanthemum Under Low Temperature Stress with Low Light

The effects of acetylsalicylic acid （ASA）, CaCl2, and ASA ＋ CaCl2 on the photosynthetic apparatus and antioxidant enzyme activities were investigated in chrysanthemum Jinba （a cut flower cultivar） under low temperature stress with low light （TL stress） （16/12℃, day/night, PFD 100 μmol m^-2 s-1）. The results showed that under TL stress, the net photosynthesis rate （Pn）, carboxylation efficiency （CE）, apparent quantum yield （AQY）, maximal photochemical efficiency （Fv/Fm） of PSII, quantum yield of PSII electron transport （ФPSII）, and photochemical quenching （qP） of the chrysanthemum leaves in all treatments were significantly decreased, but the decreases were alleviated by ASA, CaCl2, and ASA ＋ CaCl2 treatments compared with the controls. The alleviating effect of ASA ＋ CaCI2 was better than either ASA or CaCl2 single treatment. Moreover, the ASA ＋ CaCl2 treatment highly improved the chlorophyll content, relatively improved the number and size of chloroplast and starch grain in the leaves of chrysanthemum plants compared with ASA and CaCl2 treatments. It was indicated that ASA and/or CaCI2 could regulate the photosynthetic functions in the leaves of chrysanthemum plants to enhance the resistance against TL stress. On the other hand, reduction in relative conductance rate implied that ASA and/ or CaCl2 could protect from membrane injury in leaves of chrysanthemum plants. The activities of SOD, POD, and CAT in the treated leaves of chrysanthemum were increased as compared with the controls. It was suggested that ASA and/or CaCl2 had positive regulation effects on the defence enzyme activities in chrysanthemum leaves which could protect the photosynthetic apparatus to a certain degree under the TL stress. In brief, the treatment of ASA together with CaCl2 was better for chrysanthemum plants to adapt TL stress than single ASA or CaCl2 treatments.

The Role of Metal Ions in Protein and Fatty Acids Biosynthesis in Soybean under Micronutrients Application to Soil

The present study is part of our ongoing investigation to study the role of trace elements on soybean seed composition (protein, oil, and fatty acids). This study was conducted to study the effects of five trace elements (Mn, Cu, Zn, Mo, B). The treatments of Mn, Cu, Zn, Mo, and B were chlorides, except Mo as oxide, and B as boric acid. The treatments were Mn, Cu, Zn, Mo, and B alone and in combination with the chelating agent citric acid (CA), for example Mn + CA, Cu + CA, and Zn + CA. Soybean cultivar (Bolivar with maturity group V) was grown in a repeated greenhouse experiment in a randomized complete block design. The compounds were applied to three-week-old soybean plants at V3 (vegetative) and at R3 (beginning of seed-pod initiation) stages. The plants were allowed to grow until maturity under greenhouse conditions. The harvested seeds were analyzed for mineral, protein, and fatty acid contents. Results showed that Mn, Cu, and B treatments increased seed protein, while Zn, Mo, Cu + CA, and B + CA decreased the protein. Treatments of Zn, Mo, CA, Cu + CA, Zn + CA, Mo + CA, and B + CA increased the oil. Treatments of Mn and Cu decreased the oil. The Cu and B treatments increased oleic acid by 8.0% and 7.4%, respectively for Cu and B. Treatments of Mn, Mo, CA, and Mn + CA, Cu + CA, Zn + CA, Mo + CA, and B + CA decreased oleic acid by 0.6% to 14.4%. Treatments of Cu, Zn, Mo, B, CA, Mn and their combination with CA increased linoleic acid by 1.3% to 6.5%. Our goal was to identify the trace elements that would make desirable alteration in the seed composition qualities.

Extracellular and Intracellular Calcium both Involved in the Jasmonic Acid Induced Calcium Mobilization in Arabidopsis thaliana

The objective of this experiment is to evaluate the role of intracellular and extracellular Ca2＋ and calmodulin （CAM） in jasmonic acid （JA） signaling. The laser scanning microscopy was used to detect the changes of [Ca2＋]cyt of Arabidopsis thaliana leaf cells which pretreated with different types of calcium channel blocker. Moreover, the expression of VSP, one of JA response genes, was also investigated after pretreated with the above blocker and antagonist of CaM. The results showed that extracellular and intracellular calcium both involved in the JA-induced Ca2＋ mobilization, and then Ca2＋ exerted its functions through activating the CaM or CaM related proteins. The apoplast calcium influx and the calcium release from the calcium stores are both involved in the JA-induced calcium mobilization, then the JA-induced Ca2＋ transmited the JA signal through CaM or CaM related proteins, and regulated the JA responsive genes.

Ascorbic acid induces Ca2+-dependent necrosis mediated by reactive oxygen species in colorectal cancer cells

OBJECTIVE Ascorbic acid(AA),commonly known as vitamin C,is a small molecular widely distributed in in food and traditional herbs.Recently,there are some literatures reported that high concentration AA could selectively kill the cancer cells but not the normal cells.This study was designed to explore the underlying mechanisms.METHODS Colorectal cancer line cells were cultured and treated with AA.The cytotoxic,intracellular ATP level,reactive oxygen species,calcium,were determined with commercial kits and fluorescent probes.RESULTS High concentration of AA induced cell death in HCT116 and HT29 colorectal cancer cells in concentration-and time-dependent manner.AA treat⁃ment induced ATP decrease,LDH release,cell swollen and loss of plasma membrane integrity.Pharmacological inhibi⁃tors for apoptosis,necroptosis,autophagy,pyroptosis and oncosis showed no effect on AA-induced cell death.Further⁃more,ROS level increase and intracellular calcium(Ca2+)accumulation were observed after AA treatment.ROS scavenger N-acetyl cysteine(NAC),intracellular calcium chelator BAPTA-AM and intracellular calcium inhibitor 2-aminoethoxy⁃diphenyl borate(2-APB)could attenuate the cell death induced by AA.NAC could attenuate both ROS increase and intracellular Ca2+accumulation induced by AA,while BAPTA-AM could only attenuate intracellular Ca2+accumulation.In addition,high concentration AA induced mitochondrial damage and mitochondrial ROS generation.CONCLUSION AA induces Ca2+-dependent programed necrosis mediated by ROS.Our study provided new insights into high concentration AA induced cell death in human colon cancer cells.

Detection of Ca^(2+)-dependent acid phosphatase activity identifies neuronal integrity in damaged rat central nervous system after application of bacterial melanin

The study aims to confirm the neuroregenerative effects of bacterial melanin （BM） on central nervous system injury using a special staining method based on the detection of Ca^2＋-dependent acid phosphatase activity. Twenty-four rats were randomly assigned to undergo either unilateral destruction of sensorimotor cortex （group I; n = 12） or unilateral rubrospinal tract transection at the cervical level （C3-4） （group II; n = 12）. In each group, six rats were randomly selected after surgery to undergo intramuscular injection of BM solution （BM subgroup） and the remaining six rats were intramuscularly in）ected with saline （saline subgroup）. Neurological testing confirmed that BM accelerated the recovery of motor function in rats from both BM and saline subgroups. Two months after surgery, Ca^2＋-dependent acid phosphatase activity detection in combination with Chilingarian＇s calcium adenoside triphosphate method revealed that BM stimulated the sprouting of fibers and dilated the capillaries in the brain and spinal cord. These results suggest that BM can promote the recovery of motor function of rats with central nervous system injury; and detection of Ca^2＋-dependent acid phosphatase activity is a fast and easy method used to study the regeneration-promoting effects of BM on the injured central nervous system.