A Comparative Transcriptomic Study and Key Gene Targeting of Lamprey Gonadal Immune Response.

The mammalian testis and ovary possess special immunocompetence, which is central to provide protection against pathogens. However, the innate immune responses to immune challenges in lamprey gonads are poorly understood. In this study, we extracted RNA from testis and ovary tissues of lampreys at 0 hour, 8 hours and 17 days after lipopolysaccharides (LPS) stimulation and performed transcriptome sequencing. While the transcriptome profiles of the two tissues were different for the most part, genes LIP, LECT2, LAL2, GRN, ITLN, and C1q were found to be the most significantly up-regulated genes in both. Quantitative Real-time PCR (qRT-PCR) analysis confirmed that these genes were upregulated after stimulation. Furthermore, immunohistochemical staining showed that these genes in lamprey gonads are expressed in high quantities and have a specific distribution. Taken together, our results suggest that these genes could play an essential role in response of the gonads to LPS induction. This research establishes a basis for investigating the immune mechanism of vertebrate gonads and presents a fresh concept for gaining insight into the evolutionary development of jawless vertebrates.

Ultrasonic Phased Array Imaging Approach Using Omni-Directional Velocity Correction for Quantitative Evaluation of Delamination in Composite Structure.

The ultrasonic detectability of buried defects within composite materials is dependent on the anisotropy of the composite material by which the propagation property of acoustic wave in each direction is variably affected. In this study, the characteristics of acoustic waves propagating in different directions for composite materials are explored based on the full matrix capture (FMC) data using an ultrasonic phased array. The elastic constant of multidirectional carbon fiber reinforced plastic (CFRP) laminate is first derived based on the genetic algorithm. The characteristics of transmitted and reflected waves in higher angles are predicted by implementing the Christoffel equation, and the focal law used in post-processing of FMC data can be optimized accordingly. The imaging results of the total focusing method (TFM) using the improved focal law are compared with the results of the conventional TFM. The results suggest that the optimized TFM can effectively characterize the defect by reducing the background noise. Furthermore, since it is impractical to theoretically correct angle-dependent velocity for in situ inspection, a linear extrapolation method based on the experimentally measurable velocity at low angles is proposed to estimate the velocity profile at higher angles. The imaging results using the fast extrapolated velocity profile is then compared with the theoretical, and it has been demonstrated that while the difference between the images using the theoretical focal law and the linearly extrapolated one is barely visible, the later one is overwhelmingly advantageous to be realiszd for engineering practices.

Intercropping of Cyphomandra betacea with Different Ploidies of Solanum Sect. Solanum (Solanaceae) Wild Vegetables Increase Their Selenium Uptakes.

Selenium (Se) deficiency causes various diseases in humans. Se can be obtained from fruits and vegetables. In this study, the fruit tree Cyphomandra betacea was intercropped with three Solanum sect. Solanum (Solanaceae) wild vegetables [diploid (S. photeinocarpum), tetraploid (colchicine-induced S. photeinocarpum), and hexaploid (S. nigrum)], respectively, and Se uptakes of these plants were determined by a pot experiment. Intercropping decreased the biomass, photosynthetic pigment content, and superoxide dismutase activity of C. betacea, but increased the peroxidase (POD) activity, catalase (CAT) activity, and soluble protein content of C. betacea. These indicators' values of sect. Solanum increased after intercropping. The contents of Se increased in C. betacea and sect. Solanum after intercropping. Intercropped with diploid, tetraploid, and hexaploid increased the shoot Se contents in C. betacea by 13.73%, 17.49%, and 26.50%, respectively, relative to that of C. betacea monoculture. Intercropped with C. betacea increased the shoot Se contents in diploid, tetraploid, and hexaploid by 35.22%, 68.86%, and 74.46%, respectively, compared with their respective monoculture. The biomass and Se content of intercropped sect. Solanum showed linear relationships with the biomass and Se content of their monocultures. The biomass and Se content of intercropped C. betacea also exhibited linear relationships with that of sect. Solanum monocultures. Correlation and grey relational analyses revealed that the CAT activity, POD activity, and soluble protein content were the top three indicators closely associated with the C. betacea shoot Se content. The POD activity, soluble protein content, and translocation factor were the top three indicators closely associated with sect. Solanum shoot Se content. Therefore, intercropping can promote the Se uptake in C. betacea and sect. Solanum wild vegetables.

An amino acid fertilizer improves the emergent accumulator plant Nasturtium officinale R. Br. phytoremediation capability for cadmium-contaminated paddy soils.

Cadmium (Cd) contamination of paddy soil affects safe crop production. This study aimed to evaluate the effects of plant biostimulant amino acid fertilizer on the phytoremediation capability of an emergent accumulator plant Nasturtium officinale R. Br. for Cd-contaminated paddy soils. A pot study was carried out to study the effects of different concentrations of amino acid fertilizer on the Cd accumulation of N. officinale grown in Cd-contaminated paddy soil. The amino acid fertilizer increased the biomass of N. officinale. The amino acid fertilizer concentration exhibited a quadratic polynomial regression relationship with the root and shoot biomass. The fertilizer also increased the photosynthetic pigment (chlorophyll and carotenoid) contents, peroxidase (POD; EC 1.11.1.7) activity, and catalase (CAT; EC 1.11.1.6) activity of N. officinale, but decreased the soluble protein content and had no significant effect on the superoxide dismutase (SOD; EC 1.15.1.1) activity. Furthermore, the amino acid fertilizer increased the Cd content and Cd extraction of N. officinale. The shoot Cd extraction increased by 29.06%, 63.05%, 77.22%, and 17.40% at 1500-, 1200-, 900-, and 600-fold dilutions of the amino acid fertilizer, respectively, compared with the control. Moreover, the amino acid fertilizer promoted the Cd transport from the roots to shoots of N. officinale. The amino acid fertilizer concentration also exhibited a quadratic polynomial regression relationship with the root Cd content, shoot Cd content, root Cd extraction, and shoot Cd extraction, respectively. The correlation, grey relational, and path analyses revealed that the root biomass, shoot biomass, chlorophyll content, catalase activity, shoot Cd content, and root Cd extraction were closely associated with the shoot Cd extraction. Therefore, the amino acid fertilizer can promote Cd uptake and improve the phytoremediation capability of N. officinale to remediate Cd-contaminated paddy soils, and 900-fold dilution is the most suitable concentration.

An UHPLC/LC-MS illustrated the dynamic profiling of balanophorin B, gallic acid, and 4-hydroxycinnamic acid in rat as 3 molecular entities from Balanophora simaoensis.

An UHPLC/LC-MS was founded to detect balanophorin B (B), gallic acid (GA), 4-hydroxycinnamic acid (HC), and their in vivo profiling in rats, after oral administration of the ethanol extract of Balanophora simaoensis S. Y. Chang et Tam. The in vivo dynamic existence of 3 molecular entities in rats and the multistep biotransformation of GA were elucidated by their sensitive mass spectrometry response after efficient UHPLC and/or HPLC separation, through analyzing the bio-samples of rat plasma, bile, liver, kidneys, and excreta. The method was validated with satisfactory calibration curves having correlation coefficients r from 0.996 to 0.999 for concentration scaled from 0.100 nM to 0.100 µM, internal standard normalized matrix factors ranged from 0.923 to 0.993, sextuplicate recoveries valued from 95.0% to 103.6%, as well as accuracy and precision varied from 95.6% to 103.7%. The content of B, GA, and HC in the whole herb was of 4.66, 63.5, and 10.4 µmol/kg in dry weight, respectively. The Cmax for B, GA, and HC in rat systemic circulation was of 76.0 nM, 2.30 µM, and 51.0 µM, with tmax at 3, 2, and 2 h, respectively. B and GA stayed in rat liver over 4 hs to present a material base for the pharmacology and pharmacodynamics of the whole herb. The biotransformation of GA indicated a complicated scheme in rats. As a final metabolite from GA with total biotransformation conversion over 20%, 4-hydroxybenzaldehyde resourced from two steps of dehydroxylation and one step of reduction of GA, but not concerned with HC.

Effects of intercropping with Solanum photeinocarpum and its post-grafting generations on cadmium accumulation in loquat (Eriobotrya japonica).

Cadmium (Cd) contamination of orchard soils is a global problem that has been increasing. To decrease the Cd accumulation in fruits, intercropping the orchard crops with hyperaccumulator plants has been used for soil remediation. A pot and a field experiment were conducted to study the effects of intercropping the potential Cd-hyperaccumulator Solanum photeinocarpum and its post-grafting generations with loquat (Eriobotrya japonica) on the growth and Cd uptake of these two plant species. In the pot experiment, intercropping improved the biomass, Cd content, Cd extraction, and root-to-shoot Cd translocation in both species. Intercropping increased the DNA methylation levels, antioxidant enzyme activity, and soluble protein content of loquat seedlings. These results indicate that intercropping could improve the phytoremediation of S. photeinocarpum and its post-grafting generations and increase the Cd uptake in loquat seedlings. In the field experiment, intercropping increased the Cd contents in the old branches, while it decreased that in the young branches and fruits of loquat. These findings indicate that intercropping could increase the Cd uptake in old tissues but reduce the Cd uptake in young tissues and fruits of loquat. So, intercropping loquat with S. photeinocarpum and its post-grafting generations could be used in Cd-contaminated orchards.

Intercropping the potential Cd-hyperaccumulator Solanum photeinocarpum and its post-grafting generations with loquat mutually promoted the growth of two plant species, and also promoted the cadmium uptakes in S. photeinocarpum and old branches of loquat, while inhibited the Cd uptake in the loquat young tissues (young branches and fruits). These results are the new findings of the intercropping.

NIR Light-Triggered Chemo-Phototherapy by ICG Functionalized MWNTs for Synergistic Tumor-Targeted Delivery.

The combinational application of photothermal therapy (PTT), chemotherapy, and nanotechnology is a booming therapeutic strategy for cancer treatment. Multi-walled carbon nanotube (MWNT) is often utilized as drug carrier in biomedical fields with excellent photothermal properties, and indocyanine green (ICG) is a near-infrared (NIR) dye approved by FDA. In addition, ICG is also a photothermal agent that can strongly absorb light energy for tumor ablation. Herein, we explored a synergistic strategy by connecting MWNT and a kind of ICG derivate ICG-NH2 through hyaluronic acid (HA) that possesses CD44 receptor targeting ability, which largely enhanced the PTT effect of both MWNT and ICG-NH2. To realize the synergistic therapeutic effect of chemotherapy and phototherapy, doxorubicin (DOX) was attached on the wall of MWNT via π-π interaction to obtain the final MWNT-HA-ICG/DOX nanocomplexes. Both in vitro and in vivo experiments verified the great therapeutic efficacy of MWNT-HA-ICG/DOX nanocomplexes, which was characterized by improved photothermal performance, strengthened cytotoxicity, and elevated tumor growth inhibition based on MCF-7 tumor models. Therefore, this synergistic strategy we report here might offer a new idea with promising application prospect for cancer treatment.

Lamprey Immune Protein Mediates Apoptosis of Lung Cancer Cells Via the Endoplasmic Reticulum Stress Signaling Pathway.

Lamprey immune protein (LIP), a novel protein derived from the Lampetra japonica, has been shown to exert efficient tumoricidal actions without concomitant damage to healthy cells. Our study aimed to ascertain the mechanisms by which LIP inhibits lung cancer cells, thus delineating potential innovative therapeutic strategies. LIP expression in lung cancer cells was evaluated by western blotting and immunohistochemistry. Functional assays, such as high-content imaging, 3D-structured illumination microscopy (3D-SIM) imaging, flow cytometry, and confocal laser scanning microscopy, were performed to examine the proliferation and lung cancer cell apoptosis. Tumor xenograft assays were performed using an in vivo imaging system. We observed that LIP induces the decomposition of certain lung cancer cell membranes by destroying organelles such as the microtubules, mitochondria, and endoplasmic reticulum (ER), in addition to causing leakage of cytoplasm, making the maintenance of homeostasis difficult. We also demonstrated that LIP activates the ER stress pathway, which mediates lung cancer cell apoptosis by producing reactive oxygen species (ROS). In addition, injection of LIP significantly retarded the tumor growth rate in nude mice. Taken together, these data revealed a role of LIP in the regulation of lung cancer cell apoptosis via control of the ER stress signaling pathway, thus revealing its possible application in lung cancer treatment.

A TRAIL-Delivered Lipoprotein-Bioinspired Nanovector Engineering Stem Cell-Based Platform for Inhibition of Lung Metastasis of Melanoma.

Genetically engineered mesenchymal stem cells (MSCs), as non-viral gene delivery platforms, are rapidly evolving in tumor therapy due to their low immunogenicity and natural tumor-homing capacity. Methods: In this paper, we selected reconstituted high-density lipoprotein (rHDL), a lipoprotein-bioinspired nanovector with specific binding ability to scavenger receptor B type I (SR-BI) expressed on MSCs, as a transfection agent to genetically modify MSCs. pDNA encoding tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) was used as a functional gene to be transfected into the nucleus of MSCs for TRAIL expression. Lauric acid-coupled polyethyleneimine (PEI-LA) as an amphiphilic cationic polymer was synthesized to electrostatically bind to pDNA, and then incorporated into rHDL to form rHDL/PEI-LA/pDNA nanoparticles. Results: The nanoparticles exhibited homogenous particle size and excellent serum stability in vitro. Meanwhile, this SR-BI-targeted rHDL performed efficient intracellular gene delivery, specific lysosome-independent mechanism of cellular uptake and high transfection of pDNA towards MSCs. Moreover, high TRAIL expression in MSCs was detected after rHDL-mediated transfection. In vitro and in vivo results indicated that genetically engineered MSCs could accurately target to B16F10 cells, thereby producing significant apoptosis-inducing effect on aggressive melanoma. Conclusion: TRAIL-expressing MSCs engineered by rHDL nanovector was an efficient and hypotoxic method for stem cells-based pulmonary melanoma metastasis-targeting therapy.

On-Demand Versatile Prodrug Nanomicelle for Tumor-Specific Bioimaging and Photothermal-Chemo Synergistic Cancer Therapy.

Photothermal therapy is a promising approach for antitumor application although regrettably restricted by available photothermal agents. Physical entrapment of organic near-infrared dyes into nanosystems was extensively studied to reverse the dilemma. However, problems still remained, such as drug bursting and leakage. We developed here an amphiphilic prodrug conjugate by chemically modifying indocyanine green derivative (ICG-COOH) and paclitaxel (PTX) to hyaluronic acid (HA) backbone for integration of photothermal-chemotherapy and specific tumor imaging. The prepared ICG-HA-PTX conjugates could self-assemble into nanomicelles to improve the stability and reduce systemic toxicity of the therapeutic agents. The high local concentration of ICG-COOH in nanomicelles resulted in fluorescence self-quenching, leading to no fluorescence signal being detected in circulation. When the nanomicelles reached the tumor site via electron paramagnetic resonance effect and HA-mediated active targeting, the overexpressed esterase in tumor cells ruptured the ester linkage between drugs and HA, achieving tumor-targeted therapy and specific imaging. A series of in vitro and in vivo experiments demonstrated that the easily prepared ICG- HA-PTX nanomicelles with high stability, smart release behavio r, and excellent tumor targeting ability showed formidable synergy in tumor inhibition, which provided new thoughts in developing an organic near-infrared-dye-based multifunctional delivery system for tumor theranostics.

A dual-targeting reconstituted high density lipoprotein leveraging the synergy of sorafenib and antimiRNA21 for enhanced hepatocellular carcinoma therapy.

Sorafenib (So) is a multi-target kinase inhibitor extensively used in clinic for hepatocellular carcinoma therapy. It demonstrated strong inhibition both in tumor proliferation and tumor angiogenesis, while hampered by associated cutaneous side-effect and drug resistance. The knockdown of miR-21 with antisense oligonucleotides (antimiRNA21) was regarded as an efficient strategy for increasing tumor sensibility to chemotherapy, which could be employed to appreciate the efficacy of So. Herein, we successfully formulated a dual-targeting delivery system for enhanced hepatocellular carcinoma therapy by encapsulating So and antimiRNA21 in RGD pentapeptide-modified reconstituted high-density lipoprotein (RGD-rHDL/So/antimiRNA21). The RGD and apolipoprotein A-I (ApoA-I) on nanoparticles (NPs) could drive the system simultaneously to tumor neovascular and parenchyma by binding to the overexpressed ανß3-integrin and SR-B1 receptors, achieving precise delivery of therapeutics to maximize the efficacy. A series in vitro and in vivo experiments revealed that co-delivery of So and antimiRNA21 by RGD-rHDL significantly strengthened the anti-tumor and anti-angiogenic effect of So with negligible toxicity towards major organs, reversed drug-resistance and was capable of remodeling tumor environments. The constructed RGD-rHDL/So/antimiRNA21 with improved efficacy and excellent tumor targeting ability provided new idea for chemo-gene combined therapy in hepatocellular carcinoma. STATEMENT OF SIGNIFICANCE: Sorafenib (So) is a multi-target kinase inhibitor which was approved by FDA as first-line drug for hepatocellular carcinoma (HCC) therapy. However, long term application of So in clinic was hampered by serious dermal toxicity and drug resistance. Although numerous researchers were devoted to finding alternatives or therapies as combination treatments with So to reach more desired therapeutic efficacy, the therapeutic options were still limited. The present study prepares RGD pentapeptide decorated biomimic reconstituted high-density lipoprotein (rHDL) loaded with So and antimiRNA21 (RGD-rHDL/So/antimiRNA21) for enhanced HCC therapy. The RGD-rHDL/So/antimiRNA21 NPs offer an effective platform for anti-tumor and anti-angiogenesis therapy in HCC and provide new approach to reverse drug-resistance of So for feasible clinical application.

A precision-guided MWNT mediated reawakening the sunk synergy in RAS for anti-angiogenesis lung cancer therapy.

Multi-walled carbon nanotube (MWNT) with its versatility has exhibited tremendous superiority in drug delivery. Despite plenty of researches on MWNT based delivery systems, precision-guided assistances to maximize their profitable properties are still lacking in substantive progress. We developed here a dual-targeting and co-delivery system based on MWNT for antiangiogenesis therapy in lung cancer which aimed at renin-angiotensin system (RAS) dysregulation by synergistically conducting angiotensin II type 1 receptor (AT1R) and type 2 receptor (AT2R) pathway. In this work, iRGD peptide connected to polyethyleneimine (PEI) was linked to MWNT skeleton, accompanying with candesartan (CD) conjugated to MWNT mediated by cystamine (SS). The functionalized MWNT is assembled with plasmid AT2 (pAT2) to form iRGD-PEI-MWNT-SS-CD/pAT2 complexes. iRGD and CD act as pilots for complexes to dually target symbolic ανß3-integrin and AT1R both overexpressed on tumor angiogenic endothelium and lung cancer cell. CD as chemotherapy showed synergistic downregulation of VEGF when combining of pAT2 and efficiently inhibited angiogenesis. iRGD-PEI-MWNT-SS-CD/pAT2 complexes greatly appreciated drug activities by changing drug distribution and exhibited remarkable tumor growth suppression in A549 xenograft nude mice. Our work presents that such dual-targeting strategy highly improves the delivery performance of MWNT and open a new avenue for RAS related lung cancer therapy.

MicroRNA-148b Acts as a Tumor Suppressor in Cervical Cancer by Inducing G1/S-Phase Cell Cycle Arrest and Apoptosis in a Caspase-3-Dependent Manner.

BACKGROUND The purpose of our study was to investigate the role of microRNA (miR)-148b in cervical cancer. MATERIAL AND METHODS The expression of miR-148b was determined in HPV-16-immortalized cervical epithelial cell line CRL-2614 cells and in cervical cancer cell line HeLa cells. The miR-148b mimics or scrambled RNA were then transfected into Hela cells. Forty-eight hours after transfection, the mRNA expression of miR-148b and DNA methyltransferase 1 (DNMT1) were confirmed. Cell proliferation ability (cell viability and colony formation ability), invasion ability, and apoptosis were assessed after transfection with miR-148b mimics or scrambled RNA, as well as the protein expression of cyclin D1 and caspase-3. RESULTS The expression of miR-148b was significantly downregulated in HeLa cells compared with CRL2614 cells (P<0.05), but was statistically upregulated by transfection with miR-148b mimics compared with the cells transfected with scrambled RNA (P<0.05). Also, we found that the expression of DNMT1 was significantly decreased by transfection with miR-148b mimics (P<0.05). Additionally, miR-148b mimics significantly decreased the cell proliferation ability and invasion ability, and statistically induced apoptosis. Furthermore, the expression of cyclin D1 protein was significantly decreased and the expression of caspase-3 protein was significantly increased by miR-148b mimics compared with that in the cells transfected with scrambled RNA (P<0.05). CONCLUSIONS Our results suggest that overexpression of miR-148b protects against cervical cancer by inducing G1/S-phase cell cycle arrest and apoptosis through caspase-3-dependent manner, and overexpression of miR-148b might develop a therapeutic intervention for cervical cancer.

Reconstituted high density lipoprotein mediated targeted co-delivery of HZ08 and paclitaxel enhances the efficacy of paclitaxel in multidrug-resistant MCF-7 breast cancer cells.

In the past decades, reconstituted high density lipoprotein (rHDL) has been successfully developed as a drug carrier since the enhanced HDL-lipids uptake is demonstrated in several human cancers. In this paper, rHDL, for the first time, was utilized to co-encapsulate two hydrophobic drugs: an anticancer drug, paclitaxel (PTX), and a new reversal agent for P-gp (P-glycoprotein)-mediated multidrug resistance (MDR) of cancer, N-cyano-1-[(3,4-dimethoxyphenyl)methyl]-3,4-dihydro-6,7-dimethoxy-N'-octyl-2(1H)-isoquinoline-carboximidamide (HZ08). We proposed this drug co-delivery strategy to reverse PTX resistance. The study aimed to develop a biomimetic nanovector, reconstituted high density lipoprotein (rHDL), mediating targeted PTX-HZ08 delivery for cancer therapy. Using sodium cholate dialysis method, we successfully formulated dual-agent co-delivering rHDL nanoparticles (PTX-HZ08-rHDL NPs) with a typical spherical morphology, well-distributed size (~100nm), high drug encapsulation efficiency (approximately 90%), sustained drug release properties and exceptional stability even after storage for 1month or incubation in 10% fetal bovine serum (FBS) DMEM for up to 2days. Results demonstrated that PTX-HZ08-rHDL NPs significantly enhanced anticancer efficacy in vitro, including higher cytotoxicity and better ability to induce cell apoptosis against both PTX-sensitive and -resistant MCF-7 human breast cancer cell lines (MCF-7 and MCF-7/PTX cells). Mechanism studies demonstrated that these improvements could be correlated with increased cellular uptake of PTX mediated by scavenger receptor class B type I (SR-BI) as well as prolonged intracellular retention of PTX due to the HZ08 mediated drug-efflux inhibition. In addition, in vivo investigation showed that the PTX-HZ08-rHDL NPs were substantially safer, have higher tumor-targeted capacity and have stronger antitumor activity than the corresponding dosage of paclitaxel injection. These findings suggested that rHDL NPs could be an ideal tumor-targeted nanovector for simultaneous transfer of insoluble anticancer drug and drug resistance reversal agents. The PTX-HZ08-rHDL NPs co-delivery system might be a new promising strategy to overcome tumor drug resistance.

Vasodilatory effect of 14,15-epoxyeicosatrienoic acid on mesenteric arteries in hypertensive and aged rats.

The objective of this study was to investigate the 14,15-epoxyeicosatrienoic acid (14,15-EET)-induced vasodilatations as well as the underlying signaling pathways in rat mesenteric arteries from young, adult and old normotensive (WKY) and hypertensive rats. Protein expressions for prostaglandin EP(1-4) receptors, large conductance Ca(2+)-activated K(+) (BK(Ca)) channels, and adenylate cyclase (AC) were determined together with 14,15-EET-induced vasodilatations in primary- versus secondary-branches of the mesenteric artery. Responses to 14,15-EET were greater in the smaller secondary- versus primary-branches (and also more sensitive with lower EC50) and were reduced in vessels from old (80 weeks) rats as well as from vessels from the spontaneous hypertensive rats (SHR). Regardless of age or hypertension responses to 14,15-EET were inhibited by the EP2 antagonist AH6809, BK(Ca) channel inhibitor iberiotoxin, or 3',5'-cyclic monophosphate (cAMP)-protein kinase A (PKA) pathway antagonists. These data indicate 14,15-EET-induced vasodilatation is mediated via the activation of EP2 receptors and opening of BK(Ca) channels. The expressions of the EP2 receptor and AC were markedly reduced in vessels from SHR as well as old rats, whereas BK(Ca) expression was reduced in old WKY and SHR, but not adult SHR. Furthermore, expression of the p53 protein, an indicator of cell senescence and apoptosis, was elevated in adult and old SHR as well as in old WKY. In summary, attenuated 14,15-EET-induced vasodilatation in mesenteric arteries from old normotensive WKY as well as adult and old SHR is associated with reduced expression of EP2 receptors and AC.

[Effects of Schistosoma japonicum heat-shock protein 40 on macrophage activation].

OBJECTIVE: To clone and express heat-shock protein 40 gene of Schistosoma japonicum (SjHSP40) and analyze its effect on macrophage activation. METHODS: The fragment of gene encoding SjHSP40 was amplified by PCR. The gene was sub-cloned into the prokaryotic expression vector pGEX-6P-1. The recombinant plasmid pGEX-6P-1-SjHSP40 was transformed into E. coli BL21 (DE3) and induced with IPTG. The recombinant protein was purified with Glutathione-Sepharose 4B resin and analyzed by SDS-PAGE and Western-blot. The fusion protein of GST-SjHSP40 was loaded to the macrophage cell-line RAW264.7 for 48 h. Following that, the surface molecules of the macrophages were analyzed by flow cytometry. RESULTS: DNA sequencing showed that the recombinant plasmid, pGEX-6P-1-SjHSP40, was successfully constructed. The fusion protein of GST-SjHSP40 was induced, purified and specifically recognized by anti-GST antibody. Compared to GST and medium control groups, this fusion protein significantly induced the expression of co-stimulatory molecules (CD40, CD80, and CD86) and MHC-II on the surface of the macrophages. CONCLUSIONS: SjHSP40 significantly up-regulates the expression of co-stimulatory molecules and MHC-II on the surface of the macrophages. These data indicate that SjHSP40 may initiate macrophage activation.