Nanoscale dihydroartemisinin@zeolitic imidazolate frameworks for enhanced antigiardial activity and mechanism analysis.

An artificial semisynthetic material can be derived from artemisinin (ART) called dihydroartemisinin (DHA). Although DHA has enhanced antigiardial potential, its clinical application is limited because of its poor selectivity and low solubility. The drug's absorption has a direct impact on the cell, and mechanism research is limited to its destruction of the cytoskeleton. In this study, we used the zeolitic imidazolate framework-8 and loaded it with DHA (DHA@Zif-8) to improve its antigiardial potential. DHA@Zif-8 can enhance cellular uptake, increase antigiardial proliferation and encystation, and expand the endoplasmic reticulum compared with the DHA-treated group. We used RNA sequencing (RNA-seq) to investigate the antigiardial mechanism. We found that 126 genes were downregulated and 123 genes were upregulated. According to the KEGG and GO pathway analysis, the metabolic functions in G. lamblia are affected by DHA@Zif-8 NPs. We used real-time quantitative reverse transcription polymerase chain reaction to verify our results using the RNA-seq data. DHA@Zif-8 NPs significantly enhanced the eradication of the parasite from the stool in vivo. In addition, the intestinal mucosal injury caused by G. lamblia trophozoites markedly improved in the intestine. This research provided the potential of utilizing DHA@Zif-8 to develop an antiprotozoan drug for clinical applications.

Self-assembly and self-delivery of the pure nanodrug dihydroartemisinin for tumor therapy and mechanism analysis.

Dihydroartemisinin (DHA), a plant-derived natural product, has recently been proven to be an effective therapeutic agent for cancer treatment. Nevertheless, the poor water solubility and low bioavailability of DHA seriously impede its clinical applications. Herein, a simple and green strategy based on the self-assembly of DHA was developed to synthesize carrier-free nanoparticles (NPs). The resulting nanodrug (DHA NPs) was formed by the self-assembly of DHA molecules via hydrogen bonding and hydrophobic interactions. The DHA NPs exhibited a near-spherical morphology with narrow size distribution, favorable drug encapsulation efficiency (>92%), excellent stability, and on-demand drug release behavior. Furthermore, the in vitro and in vivo experiments revealed that the DHA NPs exhibited significantly higher therapeutic efficacy than the DHA equivalent. In addition, we further explored the potential molecular mechanism of the DHA NPs by utilizing RNA-seq technology and western blotting analysis, which demonstrated that the p53 signaling pathway plays a crucial part in the process of inhibiting tumor cell growth and inducing apoptosis. This work not only reveals the rationale for developing pure nanodrugs via the self-assembly of natural small molecules for oncotherapy but also the investigation of the antitumor mechanism and provides novel theoretical support for the clinical usage of DHA.

The Construction of Alkaline Phosphatase-Responsive Biomaterial and Its Application for In Vivo Urinary Tract Infection Therapy.

Urinary tract infections caused by urinary catheter implantations are becoming more serious. Therefore, the construction of a responsive antibacterial biomaterial that can not only provide biocompatible conditions, but also effectively prevent the growth and metabolism of bacteria, is urgently needed. In this work, a benzophenone-derived phosphatase light-triggered antibacterial agent is designed and synthesized, which is tethered to the biological materials using a one-step method for in vivo antibacterial therapy. This surface could kill gram-positive bacteria (Staphylococcus aureus) and gram-negative bacteria (Escherichia coli). More importantly, because this material exhibited a zwitterion structure, it does not damage blood cells and tissue cells. When the bacteria interact with this surface, the initial fouling of the bacteria is reduced by zwitterion hydration. When the bacteria actively accumulate and metabolize to produce a certain amount of alkaline phosphatase, the surface immediately started the sterilization performance, and the bactericidal effect is achieved by destroying the bacterial cell membrane. In summary, an antibacterial biomaterial that shows biocompatibility with mammalian cells is successfully constructed, providing new ideas for the development of intelligent urinary catheters.

A redox-responsive dihydroartemisinin dimeric nanoprodrug for enhanced antitumor activity.

Redox-responsive drug delivery system emerges as a hopeful platform for tumor treatment. Dihydroartemisinin (DHA) has been investigated as an innovative tumor therapeutic agent. Herein, a DHA dimeric prodrug bridged with disulfide bond as linker (DHA2-SS) has been designed and synthesized. The prepared prodrugs could self-assemble into nanoparticles (SS NPs) with high DHA content (> 90%) and robust stability. These SS NPs display sensitive redox responsive capability and can release DHA under the tumor heterogeneity microenvironment. SS NPs possess preferable antitumor therapeutic activity in contrast with free DHA. Moreover, the possible anti-cancer mechanism of SS NPs was investigated through RNA-seq analysis, bioinformatics and molecular biological method. SS NPs could induce apoptosis via mitochondrial apoptosis pathway, as well as glycolysis inhibition associate with the regulation of PI3K/AKT/HIF-1α signal path, which may offer an underlying therapeutic target for liver cancer. Our study highlights the potential of using redox responsive prodrug nanoparticles to treat cancer, meanwhile provides insights into the anti-cancer mechanism of DHA prodrug.

Global Lysine Acetylation and 2-Hydroxyisobutyrylation Profiling Reveals the Metabolism Conversion Mechanism in Giardia lamblia.

Giardia lamblia (G. lamblia) is the cause of giardiasis, a common infection that affects the general population of the world. Despite the constant possibility of damage because of their own metabolism, G. lamblia has survived and evolved to adapt to various environments. However, research on energy-metabolism conversion in G. lamblia is limited. This study aimed to reveal the dynamic metabolism conversion mechanism in G. lamblia under sugar starvation by detecting global lysine acetylation (Kac) and 2-hydroxyisobutyrylation (Khib) sites combined with quantitative proteome analyses. A total of 2999 acetylation sites on 956 proteins and 8877 2-hydroxyisobutyryl sites on 1546 proteins were quantified under sugar starvation. Integrated Kac and Khib data revealed that modified proteins were associated with arginine biosynthesis, glycolysis/gluconeogenesis, and alanine, aspartate, and glutamate metabolisms. These findings suggest that Kac and Khib were ubiquitous and provide deep insight into the metabolism conversion mechanism in G. lamblia under sugar starvation. Overall, these results can help delineate the biology of G. lamblia infections and reveal the evolutionary rule from prokaryote to eukaryote.

Preparation of T­2 toxin­containing pH­sensitive liposome and its antitumor activity.

T­2 toxin is a type A trichothecene mycotoxin. In order to reduce the side effects of T­2 toxin and increase the tumor targeting ability, a pH­sensitive liposome of T­2 toxin (LP­pHS­T2) was prepared and characterized in the present study. The cytotoxicity of LP­pHS­T2 on A549, Hep­G2, MKN­45, K562 and L929 cell lines was tested by 3­(4,5­dimethylthiazolyl­2)­2,5­diphenyltetrazolium bromide assay, with T­2 toxin as the control. The apoptotic and migratory effects of LP­pHS­T2 on Hep­G2 cells were investigated. The preparation process of LP­pHS­T2 involved the following parameters: Dipalmitoyl phosphatidylcholine: dioleoylphosphatidylethanolamine, 1:2; total phospholipid concentration, 20 mg/ml; phospholipid:cholesterol, 3:1; 4­(2­hydroxyethyl)­1­piperazineethanesulfonic acid buffer (pH 7.4), 10 ml; drug:lipid ratio, 2:1; followed by ultrasound for 10 min and extrusion. The encapsulation efficiency reached 95±2.43%. The average particle size of LP­pHS­T2 after extrusion was 100 nm; transmission electron microscopy showed that the shape of LP­pHS­T2 was round or oval and of uniform size. The release profile demonstrated a two­phase downward trend, with fast leakage of T­2 toxin in the first 6 h (~20% released), followed by sustained release up to 48 h (~46% released). From 48­72 h, the leakage rate increased (~76% released), until reaching a minimum at 72 h. When LP­pHS­T2 was immersed in 0.2 mol/l disodium phosphate­sodium dihydrogen phosphate buffers (pH 6.5), the release speed was significantly increased and the release rate reached 91.2%, demonstrating strong pH sensitivity. Overall, antitumor tests showed that LP­pHS­T2 could promote the apoptosis and inhibit the migration of Hep­G2 cells. The present study provided a new approach for the development of T­2 toxin­based anti­cancer drugs.

Supramolecular hybrids of carbon dots and dihydroartemisinin for enhanced anticancer activity and mechanism analysis.

Dihydroartemisinin (DHA) has been regarded as a potential anticancer agent in recent years. Nevertheless, the clinical applications of DHA are seriously restricted as a result of its intrinsic characteristics, such as poor water solubility, instability, and fast clearance. Herein, a type of fluorescent nanoparticles was successfully fabricated via supramolecular assembling of carbon dots (CDs) and DHA. The formulated CDs-DHA fluorescent nanoparticles not only significantly improve the solubility and stability of DHA, but also possess favorable biocompatibility and pH-dependent drug release behavior. In particular, the hybrids of CDs and DHA as nanocarriers can effectively promote the endocytosis of DHA and exhibit enhanced antitumor effects compared with free DHA in vitro and in vivo. In addition, we also explore the possible action mechanism of CDs-DHA through flow cytometric assay, transfection and western blot analysis. The results indicate that CDs-DHA nanoparticles suppress the progression of hepatic carcinoma through inducing apoptosis and inhibiting glucose metabolism, and the mechanism is related to the downregulation of PKM2 expression and the suppression of the Akt/mTOR signaling pathway, which may provide a potential therapeutic target for hepatic carcinoma treatment. This work emphasizes the great potential of utilizing CDs as a safe and convenient platform to deliver DHA for efficient cancer therapy, and the study on the anticancer mechanism can also offer theoretical support for the clinical application of DHA.

pH responsive zwitterionic-to-cationic transition for safe self-defensive antibacterial application.

Bacteria-induced infections have always been associated with various medical devices. The construction of an intelligent antimicrobial surface is an important challenge. In this study, we report the construction of a zwitterionic surface with good biocompatibility under physiological conditions and which shows an anti-adhesion effect on the original bacteria. Once the bacteria multiply, the acidic environment initiated by the bacteria will cause the amide bond on the surface to break, and the zwitterionic surface can be rapidly converted to a cationic bactericidal surface. Confocal laser scanning (CLSM) and scanning electron microscopy (SEM) show that the zwitterionic surface has efficient antibacterial activity with an anti-adhesion property while the pH-responsive transition to quaternary ammonium compounds with a germicidal surface in the acidic environment of bacterial metabolism aids the activity. Thus, the pH-responsive zwitterionic-to-cationic transition antibacterial design opens up new ideas for the efficient and safe application of cationic bactericides in clinical medical antibacterial materials.

NCOR1 may be a potential biomarker of a novel molecular subtype of prostate cancer.

Prostate cancer (PCa) is the most frequently diagnosed male cancer. An earlier study of a cohort of 333 primary prostate carcinomas showed that 74% of these tumors fell into one of seven subtypes of a molecular taxonomy defined by specific gene fusions (ERG, ETV1/4 and FLI1) or mutations (SPOP, FOXA1 and IDH1). Molecular subtypes may aid in distinguishing indolent cases from aggressive cases and improving management of the disease. However, molecular features of PCa outside the seven subtypes are still not well studied. Here we report molecular features of PCa cases without typical features of the established subtypes. We performed comprehensive genomic analysis of 91 patients, including 54 primary and 37 metastatic cases, by whole-exome sequencing. TP53, SPOP, FOXA1, AR (androgen receptor) and a TMPRSS2-ERG fusion emerged as the most commonly altered genes in primary cases, whereas AR, FOXA1, PTEN, CDK12, APC and TP53 were the most commonly altered genes in metastatic cases. Nuclear receptor corepressor (NCOR1) genomic alterations have been identified in 5% of cases, which are nontypical molecular features of PCa subtypes. A novel NCOR1 c.2182G>C (p.Val728Leu) was identified in tumor. RT-PCR was used to show that this mutation caused loss of NCOR1 exon 19 and might be oncogenic in PCa. NCOR1 is involved in maintenance of mitochondrial membrane potential in PCa cells, and loss of NCOR1 might contribute to PCa progression. Therefore, NCOR1 may be a potential molecular marker of a subtype of PCa.

MOF nanoparticles with encapsulated dihydroartemisinin as a controlled drug delivery system for enhanced cancer therapy and mechanism analysis.

Dihydroartemisinin (DHA) has attracted increasing attention as an emerging therapeutic agent for tumor treatment. However, the clinical application of DHA is seriously limited owing to its inherent properties, including low solubility, poor selectivity, and fast clearance. Herein, we report a facile yet efficient strategy based on using zeolitic imidazolate framework-8 to load DHA (DHA@ZIF-8). The as-prepared DHA@ZIF-8 nanoparticles (NPs) possess high drug encapsulation efficiency (77.2%), favorable stability, good biocompatibility and controllable drug release in tumor acidic microenvironments. DHA@ZIF-8 NPs exhibit enhanced antitumor effects compared with free DHA in in vitro and in vivo therapy experiments, accompanied with negligible side effects. Furthermore, the antitumor mechanism of DHA@ZIF-8 NPs is well investigated by RNA sequencing (RNA-seq) and bioinformatics analysis. The results indicate that DHA@ZIF-8 NPs modify the expression of 7090 genes in HepG2 cells, and the mechanism may be related to the induction of apoptosis through a p53-mediated mitochondrial pathway and the suppression of glycolysis by inhibiting the PI3K/AKT pathway. This work highlights the potential of utilizing MOFs as a safe and stable platform for developing a highly efficient drug delivery system in cancer therapy, and the investigation of the antitumor mechanism can provide theoretical support for the clinical usage of DHA.

Ac-HSP20 Is Associated With the Infectivity and Encystation of Acanthamoeba castellanii.

Acanthamoeba castellanii is a pathogenic and opportunistic free-living amoeba that causes Acanthamoeba keratitis (AK) and granulomatous amebic encephalitis (GAE) in immunocompromised individuals. The biological and pathogenic characterizations behind this opportunistic protozoan is not fully understood. This study aimed to determine the biological functions of heat shock protein (HSP)-20 of A. castellanii (Ac-HSP20) involved in the maintenance of life cycle and the infectivity of A. castellanii. Immunoscreening A. castellanii cDNA library with A. castellanii infected rabbit sera identified three positive clones, one of them was a putative heat shock protein (Ac-HSP20). The recombinant 23 kDa Ac-HSP20 protein (rAc-HSP20) was successfully expressed in Escherichia coli BL21 (DE3) and purified using metal affinity chromatography. The rabbits immunized with rAc-HSP20 produced high titer antibody (1:25,600). Immunolocalization with the antibody identified the expression of native Ac-HSP20 on the surface of both A. castellanii trophozoites and cysts. Further, Western blot with antibody identified that the expression of native Ac-HSP20 was 7.5 times higher in cysts than in trophozoites. Blocking Ac-HSP20 on the membrane of trophozoites with specific antibody or silencing Ac-hsp20 gene transcription by siRNA inhibited their transformation into cysts at the early stage but returned to normal at the late stage by stimulating the transcription of Ac-hsp20. Incubation of trophozoites with anti-Ac-HSP20 IgG increased macrophage-involved phagocytosis to the protozoa and inhibited trophozoite infectivity on the cornea of rabbits compared with that without antibody. Our study provides that Ac-HSP20 is a surface antigen involved in the encystation and infectivity of A. castellanii and thus an important target for vaccine and drug development.

Tanshinone IIA inhibits glucose metabolism leading to apoptosis in cervical cancer.

Cancer requires aerobic glycolysis to supply the energy required for proliferation. Existing evidence has revealed that blocking glycolysis results in apoptosis of cancer cells. Tanshinone IIA (Tan IIA) is a diterpenoid naphthoquinone found in traditional Chinese medicine, Danshen (Salvia sp.). Tan IIA exhibits potential anticancer activity. However, its effect on cell viability of human cervical cancer cells and its mechanism are unknown. The aim of the present study was to determine the effect of Tan IIA on proliferation and glucose metabolism in cervical cancer cells. Cell viability was measured by MTT assay, apoptosis was determined using flow cytometry and glucose uptake, lactate production, and adenosine triphosphate content were measured to assess glucose metabolism. The expression of apoptosis­associated proteins was detected by western blotting and the antitumor activity of Tan IIA in vivo was evaluated in cervical carcinoma­bearing mice. The results revealed Tan IIA treatment resulted in a considerable reduction in the viability of SiHa cells. Tan IIA decreased the expression of HPV oncogenes E6 and E7, induced apoptosis and also decreased glycolysis by suppressing the activity of the intracellular AKT/mTOR and HIF­1α. In vivo, treatment with Tan IIA resulted in a 72.7% reduction in tumor volume. The present study highlights the potential therapeutic value of Tan IIA, which functions by inducing apoptosis and may be associated with inhibition of glycolysis.

Differential distributions of parvalbumin-positive interneurons in the sulci and gyri of the adult ferret cerebral cortex.

Although accumulating evidence suggests that there are significant anatomical and histological differences between the sulci and gyri of the cerebral cortex, whether there is a difference in the distribution of interneurons between the two cortical regions remains largely unknown. In this study, we systematically compared the distributions of parvalbumin-positive interneurons among three neighboring gyrus and sulcus pairs-coronal gyrus and cruciate sulcus, anterior ectosylvian gyrus and rostral suprasylvian sulcus, and posterior ectosylvian gyrus and pseudosylvian sulcus-in the adult ferret cerebral cortex. We proposed a method to partition sulci and gyri into several specific subregions through the deepest points of the sulci and the highest points of gyri in the inner and outer cortical contours of coronal sections. We found that the density of parvalbumin-positive interneurons in the gyri was significantly higher than that in the sulci. Further study revealed that the density of PV interneurons in superficial cortical layers (layers 2/3 and layer 4) was comparable among the three pairs of sulci and gyri. However, the density of parvalbumin-positive interneurons in cortical layers 5/6 was significantly higher in gyri than in sulci. These results indicate that parvalbumin-positive interneurons are differently distributed in infragranular layers of cortical sulci and gyri.

Targeted juglone blocks the invasion and metastasis of HPV-positive cervical cancer cells.

Human papillomaviruses (HPVs), for instance, HPV 16 and HPV 18, are concerned associated with cervical cancer. Thus, it is essential to suppress HPVs-in HPV-positive cervical cancer for treating cervical cancer. The purpose of this study was to explore the proposed molecular mechanisms, which that underlies the antintumor potential of juglone to treat of HPV-positive on cervical cancer cells. The results showed that juglone suppressed HPV-positive cell growth in a dose- and time-dependent way. In addition, cell invasion and metastasis were also inhibited by juglone. Nevertheless, when pin 1 was knocked down in HPV-positive cells, cell proliferation, invasion and metastasis were reduced. This study was designed to acquire an understanding of the mechanism of invasion and metastasis in HPV-positive cells suppressed by juglone. It provides evidence of the advantageous use of juglone in the future.

Dihydroartemisinin suppresses glycolysis of LNCaP cells by inhibiting PI3K/AKT pathway and downregulating HIF-1α expression.

AIMS: Dihydroartemisinin (DHA) exhibits potential anticancer activity. However, the biological functions of DHA in prostate cancer remain largely unexplored. In this study, we aim to investigate the anti-proliferative effect and glycolysis regulation of DHA on prostate cancer cell LNCaP. MAIN METHODS: Cell proliferative activity and apoptosis inducing were detected. The gene expression was detected by mRNA microarray and results were analyzed by GO and KEGG pathway database. Expressions of glycolysis key enzymes and PI3K/AKT/HIF-1α were detected by Western blot. KEY FINDINGS: Results indicated that DHA could inhibit the LNCaP cell proliferation considerably and induce cell apoptosis. mRNA microarray showed 1293 genes were upregulated and 2322 genes were downregulated. GO and KEGG enrichment analysis suggested that glycolysis pathway was correlated with DHA inhibited the proliferation on the LNCaP cell. Western blot results showed that DHA can decrease GLUT1 and regulatory enzymes of glycolytic pathway expression probably by suppressing the activity of the intracellular Akt/mTOR and HIF-1 α. SIGNIFICANCE: Experimental validation results indicate that DHA treatment can inhibit the LNCaP cell proliferation and induce apoptosis, which may be related to glycolysis inhibition.

Development of an antibacterial surface with a self-defensive and pH-responsive function.

In this work, a charge conversion mechanism was introduced to build a bacteria responsive antibacterial surface. The antimicrobial surface is constructed by immobilizing pH-responsive moieties on the surface of the material, followed by immobilization of the antimicrobial peptide (AMP) melittin (MLT) by strong electrostatic interaction. The constructed surface exhibited self-defensive properties against Gram-positive and Gram-negative bacteria. In comparison with previously reported self-defensive systems with side effects of drug resistance, this antibacterial surface prevented the undesirable drug resistance. The bactericidal mechanism of the antibacterial surface is involved in a lytic cell membrane. Once bacteria come into contact with the surface, the bactericidal properties will be triggered on the surface. As bacteria exponentially grow, they accumulate and attach to the surface, which will develop a slightly acidic micro-environment and subsequently activate the pH responsive surface to release MLT to kill bacteria. This study broadened our understanding of the development of antibacterial surfaces and provided new insights for the antibacterial surface to be utilized in industrial, biological and medical applications.

Integrative transcriptomics, proteomics, and metabolomics data analysis exploring the injury mechanism of ricin on human lung epithelial cells.

Ricin (RT) is a plant toxin belonging to the family of type II ribosome-inactivating protein with high bioterrorism potential. Aerosol RT exposure is the most lethal route, but its mechanism of injury needs further investigation. In the present study, we performed a comprehensive transcriptomics, proteomics and metabolomics analysis on the potential mechanism of injury caused by RT on human lung epithelial cells. In total, 5872 genes, 187 proteins, and 143 metabolites were shown to be significantly changed in human lung epithelial cells after RT treatment. Molecular function, pathway, and network analyses, the genes and proteins regulated in RT-treated cells were mainly attributed to fatty acid metabolism, arginine and proline metabolism and ubiquitin-mediated proteolysis pathway. Furthermore, a comprehensive analysis of transcripts, proteins, and metabolites was performed. The results revealed the correlated genes, proteins, and metabolic pathways regulated in metabolic pathways, amino acid metabolism, transcription and energy metabolism. These genes, proteins, and metabolites involved in these dis-regulated pathways may provide a more targeted and credible direction to study the mechanism of RT injury on human lung epithelial cells. This study provides large-scale omics data that can be used to develop a new strategy for the prevention, rapid diagnosis, and treatment of RT poisoning, especially of RT aerosol.

Integration of transcriptomics, proteomics and metabolomics data to reveal the biological mechanisms of abrin injury in human lung epithelial cells.

BACKGROUND: Abrin toxin (AT) is a potent plant toxin that belongs to the type Ⅱ ribosome inactivating protein family and is recognized as an important toxin agent for potential bioweapons. Exposure to AT by way of aerosol is the most lethal route, but the mechanism of injury requires further investigation. MATERIALS AND METHODS: In the present study, we performed a comprehensive analysis of transcriptomics, proteomics and metabolomics on the potential mechanism of abrin injury in human lung epithelial cells. RESULTS: In total, 6838 genes, 314 proteins and 178 metabolites showed significant changes in human lung epithelial cells after AT treatment. Using molecular function, pathway, and network analysis, the genes and proteins regulated in AT-treated cells were mainly attributed to amino acid metabolism, lipid metabolism, and genetic information processing. Furthermore, a comprehensive analysis of the transcripts, proteins, and metabolites was performed. The results revealed that the correlated genes, proteins, and metabolism pathways regulated in AT-treated human lung epithelial cells were involved in tryptophan metabolism, biosynthesis of amino acids, and protein digestion and absorption. CONCLUSION: This study provides large-scale omics data to develop new strategies for the prevention, rapid diagnosis, and treatment of AT poisoning, especially AT from aerosol.

Juglone Inhibits Proliferation of HPV-Positive Cervical Cancer Cells Specifically.

Human papillomaviruses (HPVs), such as HPV 16 and HPV 18 are related to cervical cancer. Therefore, it is important to inhibit HPV-positive cervical cancer for treating cervical cancer. This study is aiming at investigating the proposed molecular mechanism, which underlies the antineoplastic potential of the aqueous extract of juglone of HPV-positive cervical cancer cells. According to the results, it is showed that, juglone prohibited HPV positive cervical cancer cells' growth through dose-dependent way. Nevertheless, when pin 1 was knocked down, the proliferation inhibition reduced. The detection of apoptosis and cell cycle also illustrated that juglone influenced HPV positive cells. Western blot expressed the influence mechanism that it affected the B-cell lymphoma 2 (Bcl-2) family and later activated the Caspase-depended apoptosis way. It is contributable for this study to understand the mechanism of inhibiting HPV positive cells by juglone and it also provides an effective strategy for the application of it in the future.