Unveiling the impact of CDK8 on tumor progression: mechanisms and therapeutic strategies.

CDK8 is an important member of the cyclin-dependent kinase family associated with transcription and acts as a key "molecular switch" in the Mediator complex. CDK8 regulates gene expression by phosphorylating transcription factors and can control the transcription process through Mediator complex. Previous studies confirmed that CDK8 is an important oncogenic factor, making it a potential tumor biomarker and a promising target for tumor therapy. However, CDK8 has also been confirmed to be a tumor suppressor, indicating that it not only promotes the development of tumors but may also be involved in tumor suppression. Therefore, the dual role of CDK8 in the process of tumor development is worth further exploration and summary. This comprehensive review delves into the intricate involvement of CDK8 in transcription-related processes, as well as its role in signaling pathways related to tumorigenesis, with a focus on its critical part in driving cancer progression.

Inhibition of NF-κB signaling unveils novel strategies to overcome drug resistance in cancers.

Drug resistance in cancer remains a major challenge in oncology, impeding the effectiveness of various treatment modalities. The nuclear factor-kappa B (NF-κB) signaling pathway has emerged as a critical player in the development of drug resistance in cancer cells. This comprehensive review explores the intricate relationship between NF-κB and drug resistance in cancer. We delve into the molecular mechanisms through which NF-κB activation contributes to resistance against chemotherapeutic agents, targeted therapies, and immunotherapies. Additionally, we discuss potential strategies to overcome this resistance by targeting NF-κB signaling, such as small molecule inhibitors and combination therapies. Understanding the multifaceted interactions between NF-κB and drug resistance is crucial for the development of more effective cancer treatment strategies. By dissecting the complex signaling network of NF-κB, we hope to shed light on novel therapeutic approaches that can enhance treatment outcomes, ultimately improving the prognosis for cancer patients. This review aims to provide a comprehensive overview of the current state of knowledge on NF-κB and its role in drug resistance, offering insights that may guide future research and therapeutic interventions in the fight against cancer.

High-quality wild barley genome assemblies and annotation with Nanopore long reads and Hi-C sequencing data.

Wild barley, from "Evolution Canyon (EC)" in Mount Carmel, Israel, are ideal models for cereal chromosome evolution studies. Here, the wild barley EC_S1 is from the south slope with higher daily temperatures and drought, while EC_N1 is from the north slope with a cooler climate and higher relative humidity, which results in a differentiated selection due to contrasting environments. We assembled a 5.03 Gb genome with contig N50 of 3.53 Mb for wild barley EC_S1 and a 5.05 Gb genome with contig N50 of 3.45 Mb for EC_N1 using 145 Gb and 160.0 Gb Illumina sequencing data, 295.6 Gb and 285.35 Gb Nanopore sequencing data and 555.1 Gb and 514.5 Gb Hi-C sequencing data, respectively. BUSCOs and CEGMA evaluation suggested highly complete assemblies. Using full-length transcriptome data, we predicted 39,179 and 38,373 high-confidence genes in EC_S1 and EC_N1, in which 93.6% and 95.2% were functionally annotated, respectively. We annotated repetitive elements and non-coding RNAs. These two wild barley genome assemblies will provide a rich gene pool for domesticated barley.

Genome-wide mapping of DNase I hypersensitive sites in pineapple leaves.

Pineapple [Ananas comosus (L.) Merr.] is the most economically important crop possessing crassulacean acid metabolism (CAM) photosynthesis which has a higher water use efficiency by control of nocturnal opening and diurnal closure of stomata. To provide novel insights into the diel regulatory landscape in pineapple leaves, we performed genome-wide mapping of DNase I hypersensitive sites (DHSs) in pineapple leaves at day (2a.m.) and night (10a.m.) using a simplified DNase-seq method. As a result, totally 33340 and 28753 DHSs were found in green-tip tissue, and 29597 and 40068 were identified in white-base tissue at 2a.m. and 10a.m., respectively. We observed that majority of the pineapple genes occupied less than two DHSs with length shorter than 1 kb, and the promotor DHSs showed a proximal trend to the transcription start site (>77% promotor DHSs within 1 kb). In addition, more intergenic DHSs were identified around transcription factors or transcription co-regulators (TFs/TCs) than other functional genes, indicating complex regulatory contexts around TFs/TCs. Through combined analysis of tissue preferential DHSs and genes, we respectively found 839 and 888 coordinately changed genes in green-tip at 2a.m. and 10a.m. (AcG2 and AcG10). Furthermore, AcG2-specific, AcG10-specific and common accessible DHSs were dissected from the total photosynthetic preferential DHSs, and the regulatory networks indicated dynamic regulations with multiple cis-regulatory elements occurred to genes preferentially expressed in photosynthetic tissues. Interestingly, binding motifs of several cycling TFs were identified in the DHSs of key CAM genes, revealing a circadian regulation to CAM coordinately diurnal expression. Our results provide a chromatin regulatory landscape in pineapple leaves during the day and night. This will provide important information to assist with deciphering the circadian regulation of CAM photosynthesis.

[Correlation between macrophage chemotaxis and disease severity in patients with knee osteoarthritis].

OBJECTIVE: To investigate the enhancement of macrophage chemotaxis in patients with knee osteoarthritis (KOA) and its correlation with the disease severity. METHODS: Eighty patients with KOA admitted from July 2019 to June 2022 were enrolled as the observation group and divided into 29 cases of moderate group, 30 cases of severe group and 21 cases of extremely severe group. At the same time, 30 healthy subjects were included as the control group. The gene expressions of NF-κB, CXC chemokine receptor 7 (CXCR7) and CXC chemokine ligand 12 (CXCL12) in macrophages of each group were analyzed. Visual analogue scale(VAS) was used to evaluate the degree of joint pain. Joint function was evaluated by knee Joint Society Scoring system(KSS). Finally, data analysis was carried out. RESULTS: The expression levels of NF-κB, CXCR7 and CXCL12 in moderate group, severe group and extreme recombination group were higher than those in control group. The VAS, the expression of NF-κB, CXCR7 and CXCL12 in the severe group and the extreme recombination group were higher than those in the moderate group, whereas KSS was lower than that in the moderate group. The VAS, expression levels of NF-κB, CXCR7 and CXCL12 in the extremely severe group were higher than those in the severe group, and KSS was lower than that in the severe group (all P<0.01). The expression levels of NF-κB, CXCR7 and CXCL12 in macrophages were positively correlated with VAS score, but negatively correlated with KSS(all P<0.01). The expression levels of NF-κB, CXCR7 and CXCL12 in macrophages were positively correlated with the severity of disease. After excluding the influence of traditional factors (gender, age and disease duration), multiple linear regression analysis further showed that the expression levels of NF-κB, CXCR7 and CXCL12 were still positively correlated with the severity of disease(all P<0.01). CONCLUSION: The chemotaxis of macrophages in patients with KOA increased with the aggravation of the disease, and was related to the degree of pain and function impairment.

Genome architecture and diverged selection shaping pattern of genomic differentiation in wild barley.

Divergent selection of populations in contrasting environments leads to functional genomic divergence. However, the genomic architecture underlying heterogeneous genomic differentiation remains poorly understood. Here, we de novo assembled two high-quality wild barley (Hordeum spontaneum K. Koch) genomes and examined genomic differentiation and gene expression patterns under abiotic stress in two populations. These two populations had a shared ancestry and originated in close geographic proximity but experienced different selective pressures due to their contrasting micro-environments. We identified structural variants that may have played significant roles in affecting genes potentially associated with well-differentiated phenotypes such as flowering time and drought response between two wild barley genomes. Among them, a 29-bp insertion into the promoter region formed a cis-regulatory element in the HvWRKY45 gene, which may contribute to enhanced tolerance to drought. A single SNP mutation in the promoter region may influence HvCO5 expression and be putatively linked to local flowering time adaptation. We also revealed significant genomic differentiation between the two populations with ongoing gene flow. Our results indicate that SNPs and small SVs link to genetic differentiation at the gene level through local adaptation and are maintained through divergent selection. In contrast, large chromosome inversions may have shaped the heterogeneous pattern of genomic differentiation along the chromosomes by suppressing chromosome recombination and gene flow. Our research offers novel insights into the genomic basis underlying local adaptation and provides valuable resources for the genetic improvement of cultivated barley.

Land use change impacts on red slate soil aggregates and associated organic carbon in diverse soil layers in subtropical China.

The conversion of natural forests to other land use types generally has a significant influence on soil aggregation and associated soil organic carbon (SOC) concentration, depending on soil depth. However, the dynamics underlying soil aggregate distribution and aggregate-associated SOC concentration after such conversion remain inadequately understood, especially in the red slate soil region of subtropical China, where the stability of soil aggregates is the primary deterrent to soil erosion. This study investigated the effects of land use changes on soil aggregates and aggregate-associated organic carbon content in diverse soil layers in the aforementioned region. Soil samples were collected from seven typical land use types (natural forest, artificial forest, terraced citrus orchard, downhill citrus orchard, kiwifruit orchard, cornfield, and paddy field). Sampling was conducted at a depth of 0 to 100 cm and at 20 cm increments to determine aggregate distribution and aggregate-associated SOC content. Results showed that land use change and soil depth significantly affected aggregate stability and associated SOC concentration. Upon the conversion of natural forests to orchards and croplands, both macroaggregate (>0.25 mm) and SOC concentrations decreased, thereby weakening soil resistance to erosion caused by flowing water. However, the conversion of natural forests to artificial forests did not decrease aggregate stability or aggregate-associated SOC concentration, suggesting that artificial forests are alternative tree species for soil erosion control, aggregate stability enhancement, and SOC fixation. A general linear model indicated that land use changes accounted for 55 % and 56 % of the total variations in SOC concentration in >5 mm and 2.5 mm aggregates, respectively, implying that such changes more significantly affected large-grain aggregates. This study deepens the understanding of SOC accumulation mechanisms and provides valuable information on improving soil quality and physical structure in the red slate soil region of subtropical China.

A Novel Combined Method for Measuring the Three-Dimensional Rotational Angle of a Spherical Joint.

To improve the measurement accuracy of the three-dimensional rotation angle of a spherical joint, a novel approach is proposed in this study, which combines magnetic detection by a Hall sensor and surface feature identification by an eddy current sensor. Firstly, a permanent magnet is embedded in the ball head of a spherical joint, and Hall sensors are set and distributed in the ball socket to measure the variation in the magnetic flux density when the spherical joint rotates, which are related to the 3D rotation angle. In order to further improve the measurement accuracy and robustness, we also set grooves on the ball head and use eddy current sensors to synchronously identify the rotation angle of the ball head. After the combination of two signals is performed, a measurement model is established using the RBF neural network by training, and the real-time measurement of the 3D rotation angle of the spherical joint is realized. The feasibility and superiority of this method are validated through experiments. The experimental results indicate that the measurement accuracy is substantially promoted compared to the preliminary measurement scheme based on spherical coding; the average measurement error of the single axis is reduced by 9'9″. The root mean square errors for the measurements of the 3D rotation angles in this proposed method are as follows: pitch angle α has an error of 1'8″, yaw angle ß has an error of 2'15″, and roll angle γ has an error of 29'6″.

Towards a better understanding of Pseudomonas putida biofilm formation in the presence of ZnO nanoparticles (NPs): Role of NP concentration.

Elucidating the effects of nanoparticles (NPs) on key bacterial functions not only deepens our understanding of nano-toxicity mechanisms, but also guides us in the design criteria for manufacturing safe nanomaterials. In this study, bacterial growth, biofilm development and the expression of biofilm-related genes were monitored in Pseudomonas putida KT2440, a plant-beneficial bacterium, following exposure to ZnO NPs. Low concentrations of NPs (0.5-30 mg L-1) significantly promoted bacterial growth and biofilm formation, while higher concentrations (>30 mg L-1) significantly inhibited biofilm formation. Confocal laser scanning microscopy revealed that microscope slides coated with 0.5 mg L-1 of ZnO NPs showed enhanced bacterial colonization and biomass production, but at higher concentrations (250 mg L-1), biomass production was about 11 times lower than that of the substrate without NPs. Increased protein and sugar contents of the biofilm matrix corroborated the stimulating effects of low concentrations of ZnO NPs. Physiological data were supported by changes in the expression of genes associated with oxidative stress and biofilm development. ZnO NPs at 0.5 mg L-1 stimulated the expression of quorum sensing, lipopolysaccharide biosynthesis, and antibiotic resistance genes; high concentrations of ZnO NPs (250 mg L-1) down-regulated biofilm formation-related genes and up-regulated antioxidant genes. Our results indicate that long-term release of low concentrations of ZnO NPs to the environment would promote undesired biofilm formation and increased resistance to antibiotics.

Genome-Wide Identification, Evolution, and Expression Analysis of TPS and TPP Gene Families in Brachypodium distachyon.

Trehalose biosynthesis enzyme homologues in plants contain two families, trehalose-6-phosphate synthases (TPSs) and trehalose-6-phosphate phosphatases (TPPs). Both families participate in trehalose synthesis and a variety of stress-resistance processes. Here, nine BdTPS and ten BdTPP genes were identified based on the Brachypodium distachyon genome, and all genes were classified into three classes. The Class I and Class II members differed substantially in gene structures, conserved motifs, and protein sequence identities, implying varied gene functions. Gene duplication analysis showed that one BdTPS gene pair and four BdTPP gene pairs are formed by duplication events. The value of Ka/Ks (non-synonymous/synonymous) was less than 1, suggesting purifying selection in these gene families. The cis-elements and gene interaction network prediction showed that many family members may be involved in stress responses. The quantitative real-time reverse transcription (qRT-PCR) results further supported that most BdTPSs responded to at least one stress or abscisic acid (ABA) treatment, whereas over half of BdTPPs were downregulated after stress treatment, implying that BdTPSs play a more important role in stress responses than BdTPPs. This work provides a foundation for the genome-wide identification of the B. distachyon TPS-TPP gene families and a frame for further studies of these gene families in abiotic stress responses.

Fertilization and male fertility in the rotifer Brachionus calyciflorus in the presence of three environmental endocrines.

Many studies investigated the effects of environmental endocrine disruptors with the rotifer Brachionus calyciflorus. However, they focused on the reproductive behavior of rotifers, especially male-female fertilization, as a parameter in ecotoxicological and endocrine studies. In the present study, we used two environmental hormones (progesterone and testosterone) and one nonsteroidal antiandrogen (flutamide) at five different concentrations (0.5, 1, 2, 4, and 6 mg/L) to study the reproductive behavioral parameters of male rotifers. The average swimming speed of male rotifers in the blank group was 1.14 ±â€¯0.43 mm/s. After exposure for 1 h, testosterone improved the swimming speed of males, with the greatest effect at a concentration of 2-4 mg/L, whereas flutamide and progesterone inhibited the swimming speed. Copulatory behavior experiments showed that, compared with the control group, the recognition ability of males was improved by testosterone at 1, 2, and 3 h (P < 0.05). After 4, 5, and 6 h, progesterone substantially suppressed the mating recognition ability of males, where the density of each group was extremely low at 6 h. Flutamide had a similar effect on the mating recognition ability of male rotifers as that of progesterone. The male fertilization rate in B. calyciflorus increased significantly under testosterone exposure at different concentrations (P < 0.05), with the highest level at 2 mg/L (male fertility rate = 48.61 ±â€¯3.18%). The fertilization rate of male rotifers was suppressed by both progesterone and flutamide (P < 0.05), and higher drug concentrations had stronger suppressive effects.

Effects of humic acid on the interactions between zinc oxide nanoparticles and bacterial biofilms.

The effects of humic acid (HA) on interactions between ZnO nanoparticles (ZnO NPs) and Pseudomonas putida KT2440 biofilms at different maturity stages were investigated. Three stages of biofilm development were identified according to bacterial adenosine triphosphate (ATP) activity associated with biofilm development process. In the initial biofilm stage 1, the ATP content of bacteria was reduced by more than 90% when biofilms were exposed to ZnO NPs. However, in the mature biofilm stages 2 and 3, the ATP content was only slightly decreased. Biofilms at stage 3 exhibited less susceptibility to ZnO NPs than biofilms at stage 2. These results suggest that more mature biofilms have a significantly higher tolerance to ZnO NPs compared to young biofilms. In addition, biofilms with intact extracellular polymeric substances (EPS) showed higher tolerance to ZnO NPs than those without EPS, indicating that EPS play a key role in alleviating the toxic effects of ZnO NPs. In both pure ZnO NPs and ZnO-HA mixtures, dissolved Zn2+ originating from the NPs significantly contributed to the overall toxicity. The presence of HA dramatically decreased the toxicity of ZnO NPs due to the binding of Zn2+ on HA. The combined results from this work suggest that the biofilm maturity stages and environmental constituents (such as humic acid) are important factors to consider when evaluating potential risks of NPs to ecological systems.

Effects of microcystin-producing and microcystin-freeMicrocystis aeruginosa on enzyme activity and nutrient content in the rotifer Brachionus calyciflorus.

Toxic cyanobacterial blooms disrupt freshwater recreation and adversely affect zooplankton. The freshwater cyanobacterium Microcystis aeruginosa produces microcystins, which are compounds toxic to rotifers. This study evaluated the effects of M. aeruginosa on enzyme activity and nutrient content in the rotifer Brachionus calyciflorus Pallas. The rotifers were fed on Chlorella pyrenoidosa, Scenedesmus obliquus, microcystin-producing and microcystin-free M. aeruginosa alone, and mixtures of green algae combined with toxic and nontoxic cyanobacteria, respectively. Activities of amylase, pepsase, trypsin, cellulase, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were assessed after rotifer exposure to an environmental stressor. Nutrients analyzed were glycogen, protein, and triglyceride (TG). Single cyanobacteria and mixtures combined with toxic M. aeruginosa inhibited SOD activity. CAT and GPx activities significantly increased in rotifers fed with the mixture of Chlorella and toxic cyanobacteria. The activity of digestive enzymes increased compared with the Chlorella group in single and mixed diets. Glycogen and protein decreased in Microcystis mixtures, whereas TG content increased. The grazing rate (G) of the rotifers decreased with grazing time. High G value was observed with green algae in every treatment group. Although the toxins released after grazing on Microcystis affected rotifer enzyme activity and nutrient content, B. calyciflorus changed its physiological performance and grazing intensity with food type in response to eutrophic conditions.

Life strategy and grazing intensity responses of Brachionus calyciflorus fed on different concentrations of microcystin-producing and microcystin-free Microcystis aeruginosa.

The occurrence of Microcystis blooms is a worldwide concern due to the numerous adverse effects on zooplankton. We therefore hypothesized that the cyanobacterium Microcystis aeruginosa is harmful to rotifer growth. Population and individual experiments were conducted with the same proportional volumes of Chlorella and Microcystis for given food densities. Life-table parameters, life-history traits, and the grazing intensity of Brachionus calyciflorus were evaluated after they had fed on microcystin-producing and microcystin-free Microcystis, both alone and combined with an edible alga (Chlorella pyrenoidosa), at concentrations of 1 × 105, 1 × 106, and 1 × 107 cells mL-1. The results showed that the interactive effects of food density and type appeared to be synergistic on generation time (T), net reproduction rate (R0), body length, swimming speed, and reproduction time. In contrast, these effects appeared to be antagonistic on intrinsic growth rate (r), finite rate of increase (λ), time to first brood, post-reproductive time and total offspring per female. The grazing rate of rotifers decreased with grazing time. Although the toxins released after grazing on M. aeruginosa had negative effects on rotifer growth and reproduction, B. calyciflorus changed its life strategy and grazing intensity in response to eutrophic conditions.

Metal-free inactivation of E. coli O157:H7 by fullerene/C3N4 hybrid under visible light irradiation.

Interest has grown in developing safe and high-performance photocatalysts based on metal-free materials for disinfection of bacterial pathogens under visible light irradiation. In this paper, the C60/C3N4 and C70/C3N4 hybrids were synthesized by a hydrothermal method, and characterized by X-ray diffraction (XRD), UV-vis diffuse reflection spectroscopy (UV-vis DRS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and high revolution transmission electron microscope (HRTEM). The performance of photocatalytic disinfection was investigated by the inactivation of Escherichia coli O157:H7. Both C60/C3N4 and C70/C3N4 hybrids showed similar crystalline structure and morphology with C3N4; however, the two composites exhibited stronger bacterial inactivation than C3N4. In particular, C70/C3N4 showed the highest bactericidal efficiency and was detrimental to all E. coli O157:H7 in 4h irradiation. Compared to C3N4, the enhancement of photocatalytic activity of composites could be attributed to the effective transfer of the photoinduced electrons under visible light irradiation. Owing to the excellent performance of fullerenes (C60, C70)/C3N4 composites, a visible light response and environmental friendly photocatalysts for disinfection were achieved.

Efficient Photocatalytic Disinfection of Escherichia coli O157:H7 using C70-TiO2 Hybrid under Visible Light Irradiation.

Efficient photocatalytic disinfection of Escherichia coli O157:H7 was achieved by using a C70 modified TiO2 (C70-TiO2) hybrid as a photocatalyst under visible light (λ > 420 nm) irradiation. Disinfection experiments showed that 73% of E. coli O157:H7 died within 2 h with a disinfection rate constant of k = 0.01 min(-1), which is three times that measured for TiO2. The mechanism of cell death was investigated by using several scavengers combined with a partition system. The results revealed that diffusing hydroxyl radicals play an important role in the photocatalytically initiated bacterial death, and direct contact between C70-TiO2 hybrid and bacteria is not indispensable in the photocatalytic disinfection process. Extracellular polymeric substances (EPS) of bacteria have little effect on the disinfection efficiency. Analyses of the inhibitory effect of C70-TiO2 thin films on E. coli O157:H7 showed a decrease of the bacterial concentration from 3 × 10(8) to 38 cfu mL(-1) in the solution with C70-TiO2 thin film in the first 2 h of irradiation and a complete inhibition of the growth of E. coli O157:H7 in the later 24 h irradiation.

Diagnosing x-ray power and energy of tungsten wire array z-pinch with a flat spectral response x-ray diode.

Fast z-pinch is a very efficient way of converting electromagnetic energy to radiation. With an 8-10 MA current on primary test stand facility, about 1 MJ electromagnetic energy is delivered to vacuum chamber, which heats z-pinch plasma to radiate soft x-ray. To develop a pulsed high power x-ray source, we studied the applicability of diagnosing x-ray power from tungsten wire array z-pinch with a flat spectral response x-ray diode (FSR-XRD). The detector was originally developed to diagnose radiation of a hohlraum in SG-III prototype laser facility. It utilized a gold cathode XRD and a specially configured compound gold filter to yield a nearly flat spectral response in photon energy range of 0.1-4 keV. In practice, it was critical to avoid surface contamination of gold cathode. It is illustrated that an exposure of an XRD to multiple shots caused a significant change of response. Thus, in diagnosing x-ray power and energy, we used each XRD in only one shot after calibration. In a shot serial, output of FSR-XRD was compared with output of a nickel bolometer. In these shots, the outputs agreed with each other within their uncertainties which were about 12% for FSR-XRD and about 15% for bolometer. Moreover, the ratios between the FSR-XRD and the bolometer among different shots were explored. In 8 shots, the standard deviation of the ratio was 6%. It is comparable to XRD response change of 7%.

Spermatozoon of the freshwater rotifer Brachionus calyciflorus (Rotifera, Monogononta): Advances in morphological and ultrastructural studies.

The morphological and ultrastructural features of the spermatozoon in Brachionus calyciflorus are described using light, fluorescence and transmission electron microscopy (TEM). The mature spermatozoon, which appears to be thread-like, is composed of a slightly expanded anterior of cell body region and a flagellum region without acrosome. The cell body region and flagellum region are respectively 16-27µm and 20-33µm in length (n=60). The spermatozoon is characterized by a mass of dense tubular materials, which occupy most of the cell. Some mitochondria are distributed around the nuclear region in the anterior of the cell body region, while in the posterior portion of cell body, the chromatin often contains a single lobated nucleus arranged at the center of cell. The flagellum contains the classic axoneme (9×2+2) and possesses lateral undulating membrane. Mature B. calyciflorus males have no germ cell stages earlier than the spermatids in the testis. TEM examination reveals rigid rods as well as predominant typical spermatozoon in the testis. Observations, based on successive photographs and videos, enabled a first-time recording of the unique inverted movement of the spermatozoon, which indicated that the movement of the spermatozoon is driven by the flagellum. Our study also provides further supplementary insights into the phylogenetic systematics of the Rotifera.

Calcium sensing receptor mediated the excessive generation of ß-amyloid peptide induced by hypoxia in vivo and in vitro.

Hypoxia played an important role in the pathogenesis of AD. Hypoxia increased Aß formation, then caused Alzheimer's disease. Calcium sensing receptor (CaSR) was involved in the regulation of cell growth, differentiation, hormonal secretion and other physiological function. Increasing evidence supported CaSR might play a more prominent role in susceptibility to AD, but the role of CaSR in Aß overproduction induced by hypoxia and its mechanisms remain unclear. To investigate whether CaSR mediated the overproduction of Aß induced by hypoxia, immunoblot and immunochemistry were employed to determine the expression of CaSR and BACE1 in hippocampal neurons and tissue and Ca(2+) image system was used to measure [Ca(2+)]i in hippocampal neurons. The content of Aß was detected with ELISA kits. Our research found that hypoxia increased the expression of CaSR in hippocampal neurons and tissue and [Ca(2+)]i in hippocampal neurons. Calhex 231, a selective blocher of CaSR, inhibited the increase in [Ca(2+)]i induced by hypoxia. Hypoxia or GdCl3, an agonist of CaSR, increased the expression of BACE1 in hippocampal neurons and tissue, but Calhex 231 or Xesto C (a selective inhibitor of IP3 receptor) partly prevented hypoxia-induced BACE1 overexpression. Hypoxia or GdCl3 increased the content of Aß42 and Aß40 in hippocampal tissue, however Calhex 231 or Xesto C prevented hypoxia-induced the overproduction of Aß42 and Aß40 partly. Based on the above data, we suggested that hypoxia increased [Ca(2+)]i by elevated CaSR expression to promote BACE1 expression, thereby resulting in the overproduction of Aß42 and Aß40.

[Influence of inhibited α7 nicotinic acetylcholine receptor gene expression on the production of ß-amyloid peptide in SH-SY5Y cells].

OBJECTIVE: To investigate the influence of inhibited α7 neuronal nicotinic acetylcholine receptor (nAChR) by small interference RNA (siRNA) in SH-SY5Y cells and to explore the connection of these changes with the ß-amyloid precursor protein (APP) metabolism and the pathogenesis of Alzheimer's disease (AD). METHODS: The siRNA of α7 nAChR was transfected into SH-SY5Y cells, and the expression of α7 nAChR and two subtypes of ß-secretases (BACE1 and BACE2) at mRNA and protein levels was studied by real-time PCR and Western blot, respectively. The variation of Aß(1-42) content was detected by ELISA. RESULTS: As compared with controls, the expression of α7 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with the α7 nAChR siRNA were decreased by 84% and 79% (P < 0.01), respectively. The expressions of BACE1 mRNA and protein levels was increased by 527% and 71% (P < 0.01), respectively, while the expression of BACE2 decreased by 58% and 75% (P < 0.01), respectively. The Aß(1-42) content increased by 208% (P < 0.01). CONCLUSIONS: An inhibited α7 nAChR mRNA induced by siRNA may markedly stimulate the production of Aß through the mechanism of increased expression of BACE1 and inhibited expression of BACE2, which may be related to the pathogenesis of AD.