Impacts of Human Activities and Season on Species Diversity and Abundance of Butterflies in Mpanga Kipengere Game Reserve and Surrounding Farmlands, Tanzania

Maintaining natural habitats is crucial for the preservation of insects and other species that indicate environmental changes. However, the Mpanga/Kipengere Game Reserve and its surrounding farmlands are facing disturbance due to human activities, which is putting many wildlife species, particularly larger mammals, at risk. To determine the impact of human activities on butterfly species diversity and abundance in the reserve and its surrounding areas, we conducted a study from November 2021 to October 2023. We collected butterfly data using transect walks and baited traps in two habitat types. Our study yielded 2799 butterfly Individuals ranging in 124 species divided into five families habitat, season, and anthropogenic factors are significant environmental variables influencing species diversity and abundance of butterflies. Therefore, it’s important to protect habitat and dry-season water for the conservation of invertebrates such as butterflies. Our study findings provide essential information for ecological monitoring and future assessment of the Mpanga/Kipengere Game Reserve ecosystem health.

Changes of NF-kB,p53,Bcl-2 and caspase in apoptosis induced by JTE-522 in human gastric adenocarcinoma cell line AGS cells:role of reactive oxygen species

AIM: To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process, and to investigate the changes in NF-kB, p53, bcl-2 and caspase in the apoptosis process. METHODS: Cell culture, MTT, Electromicroscopy, agarose gel electrophoresis, lucigenin, Western blot and electrophoretic mobility shift assay (EMSA) analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Lucigenin assay showed the generation of ROS in cells under incubation with JTE-522. The increased ROS generation might contribute to the induction of AGS cells to apoptosis. EMSA and Western blot revealed that NF-kB activity was almost completely inhibited by preventing the degradation of IkBalpha. Additionally, by using Western blot we confirmed that the level of bcl-2 was decreased, whereas p53 showed a great increase following JTE-522 treatment. Their changes were in a dose-dependent manner. CONCLUSION: These findings suggest that reactive oxygen species, NF-kB, p53, bcl-2 and caspase-3 may play an important role in the induction of apoptosis in AGS cells after treatment with JTE-522.

Human Disturbance Reduces Plant Species Diversity and Stability of Phyllostachys pubescens Forests

Plant species diversity is an important index reflecting the functional complexity and stability of ecosystems.Human activity can completely alter plant species diversity and cause serious degradation of ecosystems but its impact on bamboo forest still lacks of systematic evaluation.In this study,we performed a field investigation to reveal the influences of human disturbances on the plant diversity and stability of Moso bamboo forests at Southern China.The selected bamboo fields contained different intensities of human activities that could be classified as slight,moderate and severe disturbance level.Species richness index S,Shannon-Wienner index H,Simpson index D,Pielou index Jsw,community similarity index IS and community stability index were employed to quantitatively evaluate the plant species diversity and stability.The survey revealed that there were 203 species belonging to 83 families and 108 genera in Moso bamboo forests.The number of plant species in the Moso bamboo forests decreased with the increasing of disturbance intensity.The species diversity indexes generally followed the order of slight>moderate>severe disturbance,as well as the richness index S,Shannon-Wienner index H and Pielou index Jsw.The similarity and species stability of the bamboo forest communities also decreased with the increase of the disturbance intensity.Under the severe disturbance,plant species replacement occurred strongly.The obtained results provide some a guideline for the sustainable management of bamboo forest.

Spatial transcriptomics combined with single-nucleus RNA sequencing reveals glial cell heterogeneity in the human spinal cord

Glial cells play crucial roles in regulating physiological and pathological functions,including sensation,the response to infection and acute injury,and chronic neurodegenerative disorders.Glial cells include astrocytes,microglia,and oligodendrocytes in the central nervous system,and satellite glial cells and Schwann cells in the peripheral nervous system.Despite the greater understanding of glial cell types and functional heterogeneity achieved through single-cell and single-nucleus RNA sequencing in animal models,few studies have investigated the transcriptomic profiles of glial cells in the human spinal cord.Here,we used high-throughput single-nucleus RNA sequencing and spatial transcriptomics to map the cellular and molecular heterogeneity of astrocytes,microglia,and oligodendrocytes in the human spinal cord.To explore the conservation and divergence across species,we compared these findings with those from mice.In the human spinal cord,astrocytes,microglia,and oligodendrocytes were each divided into six distinct transcriptomic subclusters.In the mouse spinal cord,astrocytes,microglia,and oligodendrocytes were divided into five,four,and five distinct transcriptomic subclusters,respectively.The comparative results revealed substantial heterogeneity in all glial cell types between humans and mice.Additionally,we detected sex differences in gene expression in human spinal cord glial cells.Specifically,in all astrocyte subtypes,the levels of NEAT1 and CHI3L1 were higher in males than in females,whereas the levels of CST3 were lower in males than in females.In all microglial subtypes,all differentially expressed genes were located on the sex chromosomes.In addition to sex-specific gene differences,the levels of MT-ND4,MT2A,MT-ATP6,MT-CO3,MT-ND2,MT-ND3,and MT-CO_(2) in all spinal cord oligodendrocyte subtypes were higher in females than in males.Collectively,the present dataset extensively characterizes glial cell heterogeneity and offers a valuable resource for exploring the cellular basis of spinal cordrelated illnesses,including chronic pain,amyotrophic lateral sclerosis,and multiple sclerosis.

Effect of Human Disturbance on Species Diversity of Staphylidae

From July to September in 2005, the resources of Staphylinidae in four types of human disturbance regions （ human gathering area, grain for green re- gion, grazing region and rare disturbance region） in Erlang Mountain, Sichuan were investigated, and the individual number, species composition and diversity characteristics were analyzed. The result showed that the individual number, species diversity index （H＇） and the evenness index （J） decreased with the decrea- sing intensity of human disturbance. The species diversity index and the evenness index in human gathering area were the highest of 4.1301 6 and O. 850 2, respec- tively. Simpson predominant index （A） enhanced with the decreasing intensity of human disturbance, which reached the peak of 0. 199 6 in rare disturbance re- gion. The community similarity index/3 of Staphylinidae at different human disturbance regions ranged from 0 -0.25, indicating that the species compositions were extremely different among this four types of regions. Based on these results, it was found that human activities were closely related to individuals number and species dive^ity of Staphylinidae in Erlang Mountain, and human disturbance may add habitat heterogeneity and facilitate to the species diversity of Staphylinidae communi- ties.

Human Heterogeneity and Survival of the Species: How Did It Arise and Being Sustained?—The Conundrum Facing Researchers

Current humans, Homo sapiens, are genetically and epigenetically very heterogeneous, and subsequently also biologically and physiologically heterogeneous. Much of this heterogeneity likely arose during evolutionary processes, via various iterations of humanoid lineages, and interbreeding. While advantageous from a species perspective, the heterogeneity of humans poses serious challenges to researchers attempting to understand complex disease processes. While the use of inbred preclinical models makes the research effort more effective at some levels, the findings are often not translatable to the more heterogeneous human populations. This conundrum leads to considerable research activity with inbred preclinical models, but modest progress in understanding many complex human conditions and diseases. This article discusses several of the issues around human heterogeneity and the need to change some directions in preclinical model research. Using newer Artificial Intelligence and Machine Learning approaches can begin to deduce important elements from the complexity of human heterogeneity.

Ultraviolet light A irradiation induces immunosuppression associated with the generation of reactive oxygen species in human neutrophils

Ultraviolet blood irradiation has been used as a physical therapy to treat many nonspeci¯c diseases in clinics;however,the underlying mechanisms remain largely unclear.Neutrophils,the first line of host defense,play a crucial role in a variety of in°ammatory responses.In the present work,we investigated the effects of ultraviolet light A(UVA)on the immune functions of human neutrophils at the single-cell level by using an inverted°uorescence microscope.N-Formylmethionyl-leucyl-phenylalanine(FMLP),a classic physiological chemotactic peptide,was used to induce a series of immune responses in neutrophils in vitro.FMLP-induced calcium mobilization,migration,and phagocytosis in human neutrophils was significantly blocked after treatment with 365 nm UVA irradiation,demonstrating the immunosuppressive effects of UVA irradiation on neutrophils.Similar responses were also observed when the cells were pretreated with H2O2,a type of reactive oxygen species(ROS).Furthermore,UVA irradiation resulted in an increase in NAD(P)H,a member of host oxidative stress in cells.Taken together,our data indicate that UVA irradiation results in immunosuppression associated with the production of ROS in human neutrophils.

Role of reactive oxygen species in epithelial-mesenchymal transition and apoptosis of human lens epithelial cells

AIM:To investigate the role of reactive oxygen species(ROS)in epithelial–mesenchymal transition(EMT)and apoptosis of human lens epithelial cells(HLECs).METHODS:Flow cytometry was used to assess ROS production after transforming growth factorβ2(TGF-β2)induction.Apoptosis of HLECs after H_(2)O_(2) and TGF-β2 interference with or without ROS scavenger N-acetylcysteine(NAC)were assessed by flow cytometry.The corresponding protein expression levels of the EMT markerα-smooth muscle actin(α-SMA),the extracellular matrix(ECM),marker fibronectin(Fn),and apoptosis-associated proteins were detected by using Western blotting in the presence of an ROS scavenger(NAC).Wound-healing and Transwell assays were used to assess the migration capability of HLECs.RESULTS:TGF-β2 stimulates ROS production within 8h in HLECs.Additionally,TGF-β2 induced HLECs cell apoptosis,EMT/ECM synthesis protein markers expression,and pro-apoptotic proteins production;nonetheless,NAC treatment prevented these responses.Similarly,TGF-β2 promoted HLECs cell migration,whereas NAC inhibited cell migration.We further determined that although ROS initiated apoptosis,it only induced the accumulation of the EMT markerα-SMA protein,but not COL-1 or Fn.CONCLUSION:ROS contribute to TGF-β2-induced EMT/ECM synthesis and cell apoptosis of HLECs;however,ROS alone are not sufficient for EMT/ECM synthesis.

Diversity of Plant Species of Alpine Grassland in Nakchu of Tibet, China

Characteristics of plant species diversity of two types of grassland communities (alpine meadow and alpine grassland) was examined and the relationship between species diversity and community dynamics was determined using GPS positioning and in situscrutinization of community of alpine grassland in Nakchu prefecture of Tibet Autonomous Region. The result indicated that: ① there was an unobvious difference between grassland communities in terms of richness index of plant, evenness index, dominant index and diversity index. The species diversity index followed the order Kobresia humilis meadow >Stipa purpurea steppe>K. littledalei meadow; ② the original community created high diversity, while the degenerative community suffering from serious disturbance was of low diversity; ③ the diversity of community plants was closely related to changes of species biomass, and the growth and decline of species in the community; ④ grazing disturbance is a key factor of community dynamics, leading to coexistence of various secondary successions of communities, diversified suitable habitats and species diversity. improvements

Changes in tree species diversity and dominance across a disturbance gradient in Nepalese Sal (Shorea robusta Gaertn. f.) forests

Disturbance is often touted as a management tool, as moderate disturbance is believed to enhance diversity; thus an understanding of frequent and fluctuating disturbance regimes in forests and their effects on stand structure, dominance and diversity is very crucial. Here, the effects of different disturbance regimes, along a gradient, on diversity and dominance of five Shorea robusta Gaertn. f.-dominated forests were investigated in 25 one-ha plots in Nepal. A total of 67 tree species were recorded; of which 41 species were encountered in least disturbed and 10 species in beavily disturbed forest. Significant variations among forests were observed for all measures of alpha diversity. Alpha diversity measures declined linearly along a disturbance gradient while dominance increased linearly. Relative basal area of S. robusta increased as the level of disturbance increased, which in turn produced more than two-fold higher important value index in heavily disturbed forest than the least disturbed forest. All alpha diversity measures declined in an order of three with increasing relative basal area of S. robusta. The similarity in species composition between each pair of disturbed forests was generally low （Jaccard＇s similarity index 〈 57%）, suggesting a higher Beta diversity, It can be concluded that diversity of Sal forests declines with increasing magnitude of disturbance, which in turn favors a higher domi- nance of S. robusta. Controlling the population of the dominant species, mainly S. robusta, is recommended to enhance diversity and to achieve multiple-use forest management objectives.

Identification and interspecies characterization of UDP-glucuronosyltransferase isoforms catalyzing acacetin glucuronidation using recombinant UGT enzymes and microsomes

Objective:To explore the glucuronic acid metabolism of acacetin in human liver and intestinal microsomes to better characterize human uridine 5'-diphospho (UDP)-glucuronosyltransferase (UGT) isoforms.In addition,interspecies comparisons were performed to identify the most appropriate experimental animal model for an in vivo study.Methods:Liquid chromatography tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) were used to confirm the successful biosynthesis of acacetin-7-O-glucuronide.Human isoforms of UGT and isozyme-specific chemical inhibitors were used for recombinant assays.Acacetin glucuronidation kinetics were assessed by combining acacetin with recombinant human UGT isoforms or with microsomes from humans or experimental animals.Kinetic differences between species were assessed in vitro using the same approach.Results:We identified multiple UGT isoforms that facilitated acacetin glucuronidation,and found that UGT1A1 was the major isoform that catalyzed this process.Acacetin-7-O-glucuronide formation exhibited clear substrate inhibition kinetics when combined with recombinant UGTs or with liver/intestinal microsomes derived from humans,monkeys,rats,mice,dogs,or pigs.Intrinsic metabolic clearance values of human intestinal microsomes were two-fold greater than those of human liver microsomes.Among the evaluated species,the Km value of dog microsomes (0.86 μM) was greatest in acacetin glucuronidation,while mice exhibited the highest CLint value,5.05 mL/min/mg.The CLint values of microsomes derived from monkeys and minipigs were 1.99 mL/min/mg and 2.12 mL/min/mg,respectively,exhibiting similar intrinsic metabolic clearance activity to that observed in humans.Conclusion:Monkey may represent a suitable model for experimental studies of acacetin pharmacokinetics owing to a high sequence homology of UGT1A1 and similar UGT1A1 glucuronidation activity to humans.

Ecological constraints on elevational gradients of bird species richness in Tajikistan

The avifauna in Tajikistan has been widely studied for the last century,but specific work on species richness pattern along elevation gradients in Tajikistan is rarely investigated.Here,we reported the first study of bird species richness(BSR)in the high-altitude mountain systems(Tien Shan and Pamir-Alay)of Tajikistan which are very sensitive to the recent climate changes.We aim to explore the relationship of BSR pattern with elevation gradient and to determine the potential drivers underlying the patterns.We collected occurrence data from field surveys,published articles,and open access websites to compile a list of bird species along elevational gradients across the whole country.The BSR was counted by 100 m elevational bands ranging from 294 m to 5146 m.The patterns of BSR were calculated separately for five groups:all breeding birds,Passeriformes,Non-Passeriformes,large elevational range species,and small elevational range species.We calculated ecological and climatic factors of planimetric area,mid-domain effect(MDE),habitat heterogeneity(HH),mean annual temperature(MAT),temperature annual range(TAR),annual precipitation(AP),normalized difference vegetation index(NDVI),human influence index(HII),and human disturbance(HD)in each elevational band.A combination of polynomial regression,Pearson’s correlation,and general least squares model analyses were used to test the effects of these factors on the BSR.A unimodal distribution pattern with a peak at 750-1950 m was observed for all breeding birds.The similar pattern was explored for Passeriformes and Non-Passeriformes,while species with different elevational range sizes had different shapes and peak elevations.For all the breeding birds and Passeriformes,BSR was significantly related to spatial,climate and human influence factors,while BSR of Non-Passeriformes positively correlated with all the given factors.First,second and fourth range classes of birds were significantly correlated with human influence factors.Moreover,large-ranged species had positive correlations with the middomain effect and weakly with habitat heterogeneity.We found that area,MAT and AP were the main factors to explain the richness pattern of birds,and the species richness increases with these three factors increasing.Multiple factors such as area and climate explain 84%of the variation in richness.Bivariate and multiple regression analyses revealed a consistent influence of spatial and climate factors in shaping the richness pattern for nearly all bird groups.

Regeneration and Tree Species Diversity of Sitakund Botanical Garden and Eco-Park,Chittagong,Bangladesh

Anthropogenic disturbances influence plant regeneration and species diversity, which impact the conservation status of protected areas. A study was conducted in the Sitakund Botanical Garden and Eco-park （SBGE）, Chittagong, Bangladesh to analyze the natural regeneration and tree species diversity in disturbed and less disturbed zones. Stratified and systematic random sampling was used to select 50 sample plots from each of the two zones. A total number of 109 plant species from 43 families were recorded in the study; of which 93 species were of natural origin while the rest were planted. From the species with natural origin 66 were tree species, 9 were shrub species and 28 were climbers. Species richness, density of regeneration and disturbance index in the height classes （0- 0.5 m） and {dbh 〉 6 cm） indicated significant differences between the zones. The study analyzed how disturbances affect species diversity in the area. It was found that species richness and basal area are negatively related with disturbances. Moreover, density （N/ha） of trees was more likely to decrease with increasing tree height that reflects the huge demand of local people to harvest large trees as part of their income generating activities. The study findings have implications for future managementdecisions of the SGBE. To restore these ecosystems, management should focus on both biodiversity eonservation and providing benefits to local people without hampering forest ecosystems.

Study on the Rights to Subsistence and Development with Reference to the Origin of the Universal Declaration of Human Rights

I.The History of the Drafting of the Universal Declaration of Human Rights:the Position and Role of the UN Economic and Social Council and the General Assembly of the United Nations should not be Ignored The reason for me to use the term "source"in the discussion on the right to subsistence

The Cross-talk between ROS and p38MAPKα in the Ex Vivo Expanded Human Umbilical Cord Blood CD133^+ Cells

This study investigated the correlation between and compared the effects of reactive oxygen species（ROS） and p38 mitogen-activated protein kinase α（p38MAPKα） in the ex vivo expanded umbilical cord blood（hUCB） CD133＋ cells.hUCB CD133＋ cells were cultured in the hematopoietic stem cells（HSCs） culture medium with N-acetylcysteine（NAC,an anti-oxidant）,p38MAPKα-specific inhibitor（SB203580） or their combination.The levels of ROS and expression of phosphorylated p38MAPKα（p-p38） in CD133＋ cells were flow cytometrically detected.The efficacy of ex vivo expansion was evaluated by the density of CD133＋ cell sub-group colony-forming cells（CFC） and cobblestone area-forming cells（CAFC） assay.Our results showed decreased ROS levels in NAC,SB203580,and their combination treatment groups were almost 37%,48%,and 85%,respectively.Furthermore,SB203580 abrogated the activation of p38MAPKα more obviously than NAC.Moreover,the CD133＋ cells in SB203580 treatment group had a 21.93±1.36-fold increase,and 14.50±1.19-fold increase in NAC treatment group,but only 10.13±0.57-fold increase in control group.In addition,SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did.These findings suggested that,in expanded CD133＋ cells,ROS activates p38MAPKα,which,in turn,induces ROS production,and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.

The Chinese Expression of the Right to Subsistence:The Progressive Disclosure of the Dual Dimensions

Dedicated to the independence and prosperity of the country and the happiness of the people,the Communist Party of China has continuously enriched and developed the human rights system with Chinese characteristics during a century of arduous efforts.The right to subsistence,as the primary basic human right,is expressed from dual dimensions:the international dimension and the domestic dimension.However,the current Chinese human rights discourse hasn’t paid enough attention to the international dimension of the right to subsistence,and its content at the domestic dimension needs further clarification.This not only makes it difficult to provide a sound theoretical basis and development direction for the practices of the right to subsistence but also directly affects the integrity of the human rights system with Chinese characteristics,which is not conducive to using our human rights discourse to disseminate China’s achievements and gain international recognition.Therefore,it is vital to further improve the theory construction of the dual dimensions of the right to subsistence in the human rights system with Chinese characteristics.With the country as the main form and carrier,the international dimension of the right to subsistence originates from the people’s demands emerging from the domestic level and ending at the international level.It aims to be able to stand in the international community with dignity and possess good conditions and environments at home and abroad for the people to exercise and protect their rights.The domestic dimension of the right to subsistence centers on each individual,takes development as the measurement of human rights,and looks at the initial stage of the development axis.The progress from the domestic dimension of the right to subsistence to the right to development is achieved by satisfying the underlying needs for people’s subsistence.This also reflects the progressive deepening of emerging from the inside,responding from the outside,and then moving from the outside to the inside.

Bird communities’responses to human-modified landscapes in the southern Anhui Mountainous Area

Conversion of natural environments to human-modified landscapes is continuing at an unprecedented rate,exerting fundamental influences on global biodiversity.Understanding how wildlife communities respond to landscape modifications is critical to improve biodiversity conservation in human-dominated landscapes.In this study,we surveyed bird communities in three common habitats(i.e.,farmland,village,and forest)in the southern Anhui Mountainous Area during summer(August 2019)and winter(December 2020).The diversity metrics and species compositions of the avian communities were compared among the habitats,and the effects of land use composition in these habitats were tested.During the field surveys,we recorded 7599 birds of 120 species along 45 line transects of 1km in length.The land use compositions differed among habitats,and land use diversity was the highest in villages and lowest in forests.The species richness and bird abundance in the two human-dominated habitats(i.e.,farmland and village)were higher than those in forest in both seasons.Bird species composition also differed across habitat types in both seasons.Bird species feeding on vertebrates,fish and carrion,and species feeding on plants and seeds were mainly found in habitats with less construction lands and lower land use diversity,while omnivorous species and species feeding on fruits and nectar or on invertebrates were less affected by these two variables.The indicator species analysis showed that most species associated with forest feed on invertebrates,while species feeding on plants and seeds were more correlated with farmland and village.The results indicated that the conversion of natural habitats to human-dominated landscapes has pronounced impacts on bird communities in the study area.Human-dominated habitats harboured more avian species that deserve conservation attention.Meanwhile,bird conservations should not be relaxed in forests because there were more than 20 species that had a high specificity for forests.

TOTAL AND METHYL MERCURY LEVELS IN HUMAN SCALP HAIRS

The contents of total and methyl mercury in scalp hair samples of 1179 fishermenliving at a typical Hg-polluted region in Northeast China and 27 lying-in womenand their new born babies in Beijing have been determined by illstrumental neutronactivation analysis, gas chromatography (electron coupling) and other techniqlles.Only 18 of all fishermen have the Hg contents above 5 ig/g, which indicates that theHg pollution there has been substantially alleviated. The longitudinal Hg patternsof the lying-in women show a gradually lowering tendency during pregnant period.Further, the Hg contents of the new-born babies are generally above or close to those oftheir mothers, confirming the mechanism that the methyl Hg, an organic species of Hgwith high toxicity, is readily able to penetrate the placental barrier and accumulatedin fetus. Thus, the mercury poison hajs occurred at the early stage of pregnancy.

Experimental study of piperlongumine inducing apoptosis of human breast adenoma MDA-MB-231 cells

Objective： The aim of the study was to investigate the apoptosis induced by piperlongumine on human breast adenoma MDA-MB-231 cells and the mechanism involved. Methods： Human breast adenoma MDA-MB-231 cells line was cultured in vitro. The inhibitory effect of piperlongumine on the proliferation of human breast adenoma MDA-MB-231 cells was measured by CCK-8 assay. Distribution of cell cycle was analyzed by flow cytometry. The apoptosis rates of MDA-MB- 231 cells were measured using Annexin V/PI staining. The flow cytometry with the probe of DCFH-DA was used to detect the intracellular reactive oxygen species levels. Western blot was used to explore the protein expression of Bcl-2 and Bax. Results： The CCK-8 assay showed that piperlongumine had an inhibiting effect on the proliferation of MDA-MB-231 cells in a concentrationand time-dependent manner. MDA-MB-231 cells were markedly arrested at G0/G1 phase after treatment of piperlongumine. Piperlongumine induced apoptosis of MDA-MB-231 cells obviously. The level of intracellular reactive oxygen species was increased in a dose-dependent manner. The antioxidant N-acetyI-L-cystein inhibited the apoptosis of cells and the level of intracellular reactive oxygen species was also decreased. By Western blot analysis, we found the expression of Bax was up-regulated whereas that of Bcl-2 was down-regulated in a concentration-dependent manner. Conclusion： PiperIongumine possesses a significant function for inhibiting proliferation, arresting cells at G0/G1 phase and inducing apoptosis of MDA-MB-231 cells, which seems to be associated with the increased generation of intracellular reactive oxygen species as well as the down-regulation of Bcl-2 and up-regulation of Bax.

Detection of ROS and translocation of ERP-57 in apoptotic induced human neuroblastoma(SH-SY5Y)cells

Several toxic compounds are known to induce apoptosis in mammalian cell lines.The human neuroblastoma cells(SHSH-SY5Y)were exposed to the phosphatase inhibiting toxin okadaic acid(OA)or hydrogen peroxide(H2O2)to induce apoptosis as well as generate reactive oxygen species(ROS).Mitoxantrone(MXT)was used as a positive control for apoptosis.The SHSH-SY5Y cells were transfected with eukaryotic expression plasmid pHyPer-dMito encoding mitochondrial-targeted fluorescent or pHyPer-dCito encoding cytoplasmic-targeted fluorescent sensor for hydrogen peroxide(HyPer).The ERp57,also called GRP58(Glucose-regulated protein 58),is a stress protein induced in conditions like glucose starvation and viral infection.Recently ERp57 was shown to translocate from the endoplasmatic reticulum to the cell surface in anthracycline-induced apoptotic cells.ERp57 co-translocation together with calreticulin has been suggested to be crucial for recognizing tumor cells to induce immunogenic cell death.ERp57 translocation after exposure to okadaic acid was studied using immunofluorescence and confocal microscopy.These studies indicated that okadaic acid has induced the translocation of ERp57 to the cellular membrane.