Tumor Metabolism: A New Field for the Treatment of Glioma.

The clinical treatment of glioma remains relatively immature. Commonly used clinical treatments for gliomas are surgery combined with chemotherapy and radiotherapy, but there is a problem of drug resistance. In addition, immunotherapy and targeted therapies also suffer from the problem of immune evasion. The advent of metabolic therapy holds immense potential for advancing more efficacious and tolerable therapies against this aggressive disease. Metabolic therapy alters the metabolic processes of tumor cells at the molecular level to inhibit tumor growth and spread, and lead to better outcomes for patients with glioma that are insensitive to conventional treatments. Moreover, compared with conventional therapy, it has less impact on normal cells, less toxicity and side effects, and higher safety. The objective of this review is to examine the changes in metabolic characteristics throughout the development of glioma, enumerate the current methodologies employed for studying tumor metabolism, and highlight the metabolic reprogramming pathways of glioma along with their potential molecular mechanisms. Importantly, it seeks to elucidate potential metabolic targets for glioblastoma (GBM) therapy and summarize effective combination treatment strategies based on various studies.

Network Pharmacology Analysis and Experimental Validation to Explore the Anti-inflammatory Mechanism of Asiatic Acid on Alcoholic Steatohepatitis.

Objective: The mechanism of action of asiatic acid (AA) on alcoholic steatohepatitis (ASH) was investigated using network pharmacology and experiments. Methods: Through data retrieval, network construction, and enrichment analysis, the potential mechanism of AA in the treatment of alcoholic steatohepatitis was explored. Animal and cell models were established in this study. Animal Model. The mouse model was divided into six groups: normal group; model group; low, medium, and high AA group; and silibinin-positive group. Cell Model. An in vitro inflammatory model of RAW264.7 cells was established by alcohol stimulation. Results: Compared with the model group, the low, medium, and high AA group showed decreased serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and total cholesterol (T-CHO). The inflammatory factor tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1ß), and interleukin 6 (IL-6) in a dose-dependent manner were decreased. In addition, hematoxylin-eosin staining showed that liver tissue damage and inflammatory cell infiltration in mice were significantly reduced with increasing doses. Further, oil red staining showed that lipid accumulation in hepatocytes in the low, medium, and high AA group was significantly reduced, with increasing dose. In addition, in the cellular model, real-time reverse transcriptase-polymerase chain reaction (Real-Time RT-PCR) and enzyme-linked immunosorbent assay (ELISA) results showed that AA could alleviate alcohol-induced cellular inflammation, while western blot and immunofluorescence results showed that AA might alleviate alcohol-induced cellular inflammation by inhibiting the nuclear factor-κB (NF-κB) pathway. Conclusion: This study provides multiple lines of evidence that asiatic acid may alleviate alcoholic hepatitis in mice by modulating the NF-κB pathway.

Particle Design and Inhalation Delivery of Iodine for Upper Respiratory Tract Infection Therapy.

Diseases caused by upper respiratory tract (URT) and pulmonary infections have been a serious threat to human health for millennia and lack of targeted effective therapeutic techniques. In this study, two kinds of cyclodextrin particles with typical particle shapes of nanocubes and microbars were synthesized through a facile process. Subsequently, the particles were used as carriers for loading and stabilizing iodine and characterizations were performed to demonstrate the loading mechanism. Next-generation impactor (NGI) experiments showed that iodine-loaded microbars (I2@microbars) had a deposition rate of 79.75% in URT, while iodine-loaded nanocubes (I2@nanocubes) were delivered to the deep lungs with a fine particle fraction (FPF) of 46.30%. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) indicated that the iodine-loaded nanocubes and microbars had similar bactericidal effect to povidone iodine solution. Cell viability studies and extracellular pro-inflammatory factor (TNF-α, IL-1ß, IL-6) evaluations demonstrate noncytotoxic effects of the blank carriers and anti-inflammatory effects of iodine-loaded samples. The irritation of the rat pharynx by I2@microbars was evaluated for the behavioral observations, body weight changes, histopathological studies, and TNF-α, IL-1ß, and IL-6 levels in pharyngeal tissues. The results showed that I2@microbars had no irritation to rat pharyngeal tissues at therapeutic doses. In conclusion, the present study provides novel treatment of URT infections via supramolecular cyclodextrin carriers for URT local therapy with iodine loading by a solvent-free method, which enhances the stability and reduces the inherent irritation without inhibiting their antimicrobial effects. Two kinds of cyclodextrin particles with typical shapes of microbars and nanocubes were synthesized by a facile process. Subsequently, iodine was successfully loaded into the particles by gas-solid interaction. The iodine-loaded microbars showed air dynamics characteristics for inhalation delivery to the upper respiratory tract with little alveolar deposition in the lungs.

Physical disorder and crime revisited: New evidence from intensive longitudinal data.

OBJECTIVES: The purpose of this study is to investigate whether the often-debated association between physical disorder and crime exists longitudinally and operates through a specific spatial process. METHODS: We combine four administrative and official databases to analyze patterns of crime occurring each month in New Orleans, LA, between 2012 and 2018 at the block group level. We adopt a generalized additive model (GAM) framework to efficiently account for potential spatial, temporal, and spatiotemporal autocorrelation to investigate this association. RESULTS: The resident-driven self-report measure of physical disorder (i.e., 311 calls) consistently predicts neighborhood criminal activities in New Orleans. The spatial effects of 311 calls in adjacent areas on drug violations, property crime, and violence in a focal neighborhood are also observed. In contrast, official reports of physical disorder, captured with code violation, is either weakly related to crime or lacking an empirical relationship. CONCLUSION: Overall, our findings suggest that the association between physical disorder and neighborhood crimes is robust over time. In addition, this association is conditioned by how physical disorder is operationalized, with dimensions such as residential report and official report of physical disorder playing a unique, and occasionally important role in understanding overall patterns of neighborhood criminal activity.

Children's immigrant generational status and delinquency: The mediating effects of friendship networks.

A mounting body of empirical studies demonstrates that first-generation immigrant children have a lower level of delinquency and crime but second and third-plus generations report a precipitous increase in these behaviors. Adopting a social network approach, we analyzed the behavioral and structural characteristics of children's friendship networks across the first, second, and third-plus immigrant generations, and investigated the mediating role of these friendship traits in explaining generational disparity. Our results reveal that children's friendship networks differ in structural (e.g., popularity) and behavioral features (e.g., network deviance) across immigrant generations. These friendship features, particularly network peer deviance, the percentage of second-generation friends, and children's popularity mediate the association between immigrant generational status and children's delinquency. Extending previous research, our study highlights the importance of applying the social network approach to understand delinquency disparity across immigrant generations and suggests that the composition of friendship networks play an important role in immigrant children's delinquency involvement.

Formulation and In Vitro Release Kinetics of Mucoadhesive Blend Gels Containing Matrine for Buccal Administration.

Enterovirus 71 (EV71) is a pathogenic factor of severe hand, foot, and mouth disease (HFMD). No vaccine or specific treatment is currently available for EV71 infection. Hence, we developed a buccal mucoadhesive gel containing matrine to protect against HFMD. Mucoadhesive gels were prepared by Carbopol 974P and were combined with Carbopol 971P, sodium carboxymethyl cellulose (CMC-Na), or hydroxypropylmethy cellulose (HPMC K100M). The formulations were characterized in terms of tensile testing and continuous flow techniques for mucoadhesion. The rheological studies and in vitro drug release characteristics were also investigated. The results showed that combinations of two polymers significantly improved mucoadhesion, especially Carbopol 974P blended with HPMC. Carbopol 974P to HPMC blend ratios of 1:1 and 2:1 induced better mucoadhesion in the tensile test and continuous flow method, respectively. The most sustained release was obtained at a Carbopol 974P to HPMC ratio of 2.5:1. A predominantly non-Fickian diffusion release mechanism was obtained. The gel containing 2.5% Carbopol 974P combined with 1% HPMC showed good mucoadhesion properties and sustained drug release.

Delivery of risperidone from gels across porcine skin in vitro and in vivo in rabbits.

The purpose of this study was to develop and evaluate a transdermal delivery system for RIS using hydrogels. First, the effects of different concentrations of hydroxypropyl methylcellulose and Carbomer 934 (CBR) on RIS permeation were investigated in porcine skin. The optimized formulation was chosen as the base gel to screen for penetration enhancers. The pharmacokinetics of the optimized RIS formulation was then studied in vitro in rabbits. A formulation with 0.5% CBR showed the highest RIS permeation and was selected as the base gel. RIS permeation was further increased by incorporation of Azone, lauryl alcohol, or menthol, and the enhancing effects of the three were dose-dependent. When each enhancer combined with propylene glycol (PG) a synergistic effect was found. A combination of 6% menthol and 6% PG exhibited highest RIS in vitro penetration rate and showed a high efficiency in vivo, with a relative bioavailability of 131.53% compared with intragastric administration. These findings showed that 1% RIS in 0.5% CBR, containing a combination of 6% menthol and 6% PG, can deliver doses of RIS that are therapeutically relevant for treating patients with schizophrenia.

Development and Evaluation of a Novel Hyaluronic Acid and Chitosan-modified Phytosome for Co-delivery of Oxymatrine and Glycyrrhizin for Combination Therapy.

BACKGROUND: Multidrug resistance (MDR) of cancer cells is a major obstacle to efficient cancer chemotherapy. Combination therapy is expected to enhance the anticancer effect and reverse MDR. Numerous patents involve different kinds of nanoparticles for the co-delivery of multiple chemotherapeutics, but the FDA has approved none. OBJECTIVE: In this study, oxymatrine (OMT) and glycyrrhizin (GL) were co-loaded into phytosomes as the core of nanocarriers, and the shell was cross-linked with chitosan (CS) and hyaluronic acid (HA) with the capability for the controlled, sequential release and the targeted drug uptake. METHODS: Phospholipid complexes of OMT and GL (OGPs) were prepared by a solvent evaporation technique and could self-assemble in an aqueous solution to form phytosomes. CS and HA were sequentially coated on the surface of OGPs via electrostatic interactions to obtain CS coated OGPs (CS-OGPs) and HA modified CS-OGPs (HA-CS-OGPs), respectively. The particle size and zeta potential were measured to optimize the formulations. In vitro cytotoxicity and cellular uptake experiments on HepG2 cells were performed to evaluate the anticancer activity. RESULTS: OGPs were obtained with nano-size around 100 nm, and CS and HA coating on phytosomes could change the particle size and surface potential. The drug loading of OMT and GL showed that the nanocarriers could maintain a fixed ratio of 1:1. The in vitro release experiments indicated the release of OMT and GL was pH-dependent and sequential: the release of OMT from CS-OGPs and HA-CS-OGPs was significantly increased at pH 5.0 compared to the release at pH 7.4, while GL exhibited sustained released from CS-OGPs and HA-CS-OGPs at pH 5.0. Furthermore, in vitro cytotoxicity and cellular uptake experiments on HepG2 cells demonstrated that the co-delivery system based on phytosomes had significant synergistic anti-tumor activities, and the effects were enhanced by CS and HA modification. CONCLUSION: The delivery of OMT and GL via HA-CS-OGPs might be a promising treatment to reverse MDR in cancer therapy.

The basic leucine zipper transcription factor MeaB is critical for biofilm formation, cell wall integrity, and virulence in Aspergillus fumigatus.

The regulation of fungal cell wall biosynthesis is crucial for cell wall integrity maintenance and directly impacts fungal pathogen virulence. Although numerous genes are involved in fungal cell wall polysaccharide biosynthesis through multiple pathways, the underlying regulatory mechanism is still not fully understood. In this study, we identified and functionally characterized a direct downstream target of SomA, the basic-region leucine zipper transcription factor MeaB, playing a certain role in Aspergillus fumigatus cell wall integrity. Loss of meaB reduces hyphal growth, causes severe defects in galactosaminogalactan-mediated biofilm formation, and attenuates virulence in a Galleria mellonella infection model. Furthermore, the meaB null mutant strain exhibited hypersensitivity to cell wall-perturbing agents and significantly alters the cell wall structure. Transcriptional profile analysis revealed that MeaB positively regulates the expression of the galactosaminogalactan biosynthesis and ß-1,3-glucanosyltransferase genes uge3, agd3, and sph3 and gel1, gel5, and gel7, respectively, as well as genes involved in amino sugar and nucleotide sugar metabolism. Further study demonstrated that MeaB could respond to cell wall stress and contribute to the proper expression of mitogen-activated protein kinase genes mpkA and mpkC in the presence of different concentrations of congo red. In conclusion, A. fumigatus MeaB plays a critical role in cell wall integrity by governing the expression of genes encoding cell wall-related proteins, thus impacting the virulence of this fungus.IMPORTANCEAspergillus fumigatus is a common opportunistic mold that causes life-threatening infections in immunosuppressed patients. The fungal cell wall is a complex and dynamic organelle essential for the development of pathogenic fungi. Genes involved in cell wall polysaccharide biosynthesis and remodeling are crucial for fungal pathogen virulence. However, the potential regulatory mechanism for cell wall integrity remains to be fully defined in A. fumigatus. In the present study, we identify basic-region leucine zipper transcription factor MeaB as an important regulator of cell wall galactosaminogalactan biosynthesis and ß-1,3-glucan remodeling that consequently impacts stress response and virulence of fungal pathogens. Thus, we illuminate a mechanism of transcriptional control fungal cell wall polysaccharide biosynthesis and stress response. As these cell wall components are promising therapeutic targets for fungal infections, understanding the regulatory mechanism of such polysaccharides will provide new therapeutic opportunities.

A Qualitative Assessment of Social Norms Related to Seeking Help for Intimate Partner Violence in Honduras.

Intimate partner violence (IPV) is a major public health issue in Honduras and other low- and middle-income countries, with few victims seeking help. While structural factors, such as lack of services and economic barriers, are often cited as reasons for not seeking help, social and cultural factors may also play a role. This study aims to describe the normative social environment that may hinder women's help-seeking behaviors for IPV. Thematic analysis was conducted on data from four focus group discussions with 30 women at a busy health center in urban Tegucigalpa, Honduras. Data were coded inductively and themes were identified deductively using the theory of normative social behavior and its components (descriptive and injunctive social norms, expected outcomes, and groups of reference). Four themes emerged: social norms and expected outcomes that discourage IPV help-seeking; factors that determine the direction of a social norm, either discouraging or encouraging help-seeking; groups of reference for IPV victims; and society sets women up for failure. Social norms, expected outcomes, and groups of reference hinder women's help-seeking behavior after IPV. These findings have significant implications for designing effective interventions and policies to support women and their families affected by IPV.

Identification of a conserved linear B-cell epitope in the M protein of porcine epidemic diarrhea virus.

BACKGROUND: The major structural protein of coronaviruses, the membrane (M) protein, can elicit the formation of protective antibodies, but little information is available about the M protein of porcine epidemic diarrhea virus (PEDV). Identification of epitopes on the PEDV M protein will be helpful in the elucidation of the antigenic properties of this protein. RESULTS: One hybridoma cell line secreting anti-M protein monoclonal antibody (McAb) was generated and designated 4D4. To map the epitopes on the PEDV M protein, a total of 17 partially overlapping fragments covering the C-terminus of M protein were expressed as fusion proteins with a 6×His tag or a GST tag. A linear motif, 193TGWAFYVR200, was identified by enzyme-linked immunosorbent assay (ELISA) and western blot (WB) analysis using McAb 4D4. The motif 195WAFYVR200 was the minimal requirement for reactivity, as demonstrated by removing amino acids individually from both ends of the motif 193TGWAFYVR200. The result of WB analysis showed that the 4D4-defined epitope could be recognized by PEDV-positive serum, but not transmissible gastroenteritis virus (TGEV)-positive serum. Furthermore, this epitope was highly conserved among different PEDV strains, as shown by alignment and comparison of sequences. CONCLUSION: A McAb, 4D4, directed against the M protein of PEDV, was obtained, and the 4D4-defined minimal epitope sequence was 195WAFYVR200. The McAb could serve as a candidate for development of a McAb-based antigen capture ELISA for detection of PEDV. The epitope identified provides a basis for the development of epitope-based differential diagnostic techniques and may be useful in the design of epitope-based vaccines.

Impact of running away on girls' pregnancy.

This study assessed the impact of running away on pregnancy in the subsequent year among U.S. adolescents. We also investigated interactions between running away and sexual assault, romance, and school disengagement. Pregnancy among females between 11 and 17 years (n = 6100) was examined utilizing the Longitudinal Study of Adolescent Health (Add Health) data from Waves 1 and 2. The odds of pregnancy in the next year were 1.67 times greater for runaways net of other factors. A history of sexual assault and romantic involvement increased the likelihood of pregnancy. The relationship between pregnancy and runaway behavior in the general population is understudied. Our findings suggest that runaway youth have a multiplicity of needs that require a complex array of medical, social, emotional, and academic resources to promote positive sexual health outcomes.

Compulsive immobility: Understanding the role of health on collective efficacy.

Most health care approaches to understanding social ills are rooted in strain or ecological models. Strain models assume that the impact of poor physical health operates through the individual, that it is the individual suffering from poor health who engages in social ills as a means of adapting, and that the impact of poor health is rather direct and immediate. Meanwhile, ecological approaches of health acknowledge how poor health may impact others and the collective, but poorly account for the case in which this is not so, leaving unexplained the many instances of people who are in poor health but remain actively engaged with their communities and preserve relationships that nurture trust, shared norms, and cooperation. To rectify this problem, we introduce the concept of "compulsive immobility": the situation in which those in poor health are compelled to stay indoors and refrain from community socialization. We argue that compulsive immobility mediates the relationship between poor physical health and collective efficacy, suggesting that illness, specifically to a point of physical immobility (e.g., bedridden), enables poor health to detract from collective efficacy. This allows scholars to both acknowledge how poor health may impact the individual and community, while specifying the mechanism through which it operates. To support our claim, we draw on GSS data to examine the relationship among poor health, health-related immobility, and collective efficacy. Our results provide empirical support for our argument, revealing that general health conditions influenced the level of generalized trust directly and indirectly through compulsive immobility. We conclude with suggestions on how compulsive immobility might impact neighborhood crime and propose ways through which subsequent research may refine and further test compulsive immobility as a mediator between poor health and collective efficacy.

Self-Control, Risky Lifestyles, and Victimization among Chinese Adolescents.

Violent and property victimization among Chinese adolescents remains a social problem, yet studies that incorporate individual characteristics and situational/contextual factors to explain such victimization remain scarce. Drawing upon survey data collected from a large, representative sample of middle school students from two areas in Guizhou Province, China, we test Schreck's integrated model of victimization, finding that self-control has both direct and indirect influences on violent and property victimization among Chinese adolescents. Delinquent peers play the most significant intermediate role in connecting self-control and adolescent victimization. Results reconfirm the importance of both self-control and risky lifestyles/situations in shaping victimization, and identify a victimization pathway that accentuates the key linking mechanism of delinquent peers in the self-control-victimization nexus.

Parental Migration and Children's Exposure to Polyvictimization in Rural China.

The present study aims to investigate (1) the difference in polyvictimization rate between children left behind and those living with non-migrant parents in rural China, and (2) the social processes through which the effects of parental migration are conveyed through children's behaviors and interaction with the immediate external environment (e.g., family, school, and peers). The research hypotheses were tested by using a probability sample consisting of 1,681 middle school students in rural areas in Guizhou province (Mean age = 13.55, SD = 1.01; 50% of the participants were boys). Findings from multivariate logistic regression models and indirect effect analyses suggest that (1) overall, left-behind children are exposed to a higher level of polyvictimization than children living with both non-migrant parents in rural China and (2) whereas all left-behind children face similar challenges in school and family settings, each type of parental migration and caretaking arrangement entails unique protective or risk factors of polyvictimization. As one of the first studies to systematically investigate the rate and etiology of polyvictimization among children left behind in rural China, this study highlights the prominent role of parental migration in reproducing and reinforcing children's differential exposure to polyvictimization among China's rural families.

Development and Evaluation of a Novel Diammonium Glycyrrhizinate Phytosome for Nasal Vaccination.

The objective of the present research was to formulate diammonium glycyrrhizinate (DG) into phytosomes (DG-P) to induce nasal immune responses and enhance absorption. Plackett- Burman design was used for process optimization, incorporating specific formulation and process variables to obtain the optimal parameters. Fourier transform infrared spectroscopy (FTIR), X-ray power diffraction (P-XRD), and transmission electron microscopy (TEM) were used for characterization. The adjuvant activity of the DG-P was evaluated by using bone marrow dendritic cells. In vitro nasal mucosal permeation and in situ nasal perfusion were also investigated to evaluate nasal absorption. The DG phytosomes were in the size range of 20~30 nm and zeta-potential range of -30~-40 mV. DG-P demonstrated 4.2-fold increased solubility in n-octanol. Coculturing bone marrow dendritic cells with DG-P led to enhanced dendritic cell maturation. Apparent permeability coefficient of the phytosomal formulation was almost four times higher than that of free DG determined by ex vivo permeation studies on excised porcine mucosa. In situ nasal perfusion studies in rats demonstrated that the nasal absorption of DG-P was significantly higher than that of free DG. Conclusively, the results confirmed that DG-P have potential for use as an adjuvant for nasal vaccine.

Neuroprotective effects of CysLTR antagonist on Streptococcus pneumoniae-induced meningitis in rats.

Cysteinyl leukotrienes (CysLTs) modulate central nervous system inflammatory responses via their receptors, CysLT1R and CysLT2R. It has been demonstrated that CysLTR participates in the infection process of Streptococcus pneumoniae (SP)-induced meningitis. In the present study, the effects and possible underlying mechanisms of CysLTR antagonists (pranlukast and HAMI 3379) on SP meningitis were further determined. SP meningitis was induced by intracerebroventricular injection of serotype III SP in Sprague-Dawley rats which were administrated intraperitoneally with 0.1 mg/kg antagonists. The clinical disease status of rats was evaluated by body weight and behavioral changes with neurological scoring. Survival neuron density, activated microglial and astrocytes were assessed by Nissl staining and immunohistochemical staining. The expression levels of inflammatory cytokines and NLRP3 inflammasome were detected by reverse transcription-quantitative PCR and western blotting, respectively. Pranlukast and HAMI 3379 treatment markedly alleviated the clinical disease status, which was manifested by improving body weight loss and neurological deficit. Furthermore, pranlukast and HAMI 3379 treatment ameliorated neuronal injury and inhibited microgliosis and astrogliosis. In addition, significant downregulation of inflammatory cytokines and NLRP3 expression was observed in pranlukast and HAMI 3379-treated rats. These in vivo findings indicated the neuroprotective effects of CysLTR antagonists against experimental SP-induced meningitis, and the mechanism of anti-inflammatory effects may partly be by inhibiting NLRP3 inflammasome overactivation.

Efficient capture and stabilization of iodine via gas-solid reaction using cyclodextrin metal-organic frameworks.

For treatment of wound infection with stabilized iodine, potassium iodide cyclodextrin metal-organic frameworks (KI-CD-MOF) was prepared to carry iodine via gas-solid reaction. Apart from highly ordered porous frameworks, KI-CD-MOF contains uniformly distributed iodide ions which stabilize iodine (I2). X-ray photoelectron spectroscopy and potentiometric titration were utilized to confirm the formation of I3- in the highly porous KI-CD-MOF as I2@KI-CD-MOF. Molecular simulation and characterizations of the synchrotron radiation Fourier transform infrared spectroscopy, differential scanning calorimeter, powder X-ray diffraction and N2 adsorption were conducted to illustrate the inclusion mechanism of iodine in I2@KI-CD-MOF. The apparent solubility of iodine in water was 3.86 times enhanced. The stability and antibacterial activity tests demonstrated that the highly-dispersed iodide ions in KI-CD-MOF are crucial in improvement to the solubility, stability, and bacteriostatic effects of active iodide. Therefore, KI-CD-MOF has broad application prospects and advantages in efficiently capturing and stabilizing iodine.

The influence of running away on the risk of female sexual assault in the subsequent year.

This study explores the sexual risk trajectories of female youths and sheds light on the long-term effects of running away. It evaluates whether running away increases the risk of sexual assault in the following year, which is after runaways return home. The sample consists of 5,387 heterosexual females between the ages of 11 and 18 years from the Longitudinal Study of Adolescent Health. Nearly one quarter (23%) of runaways report a previous sexual assault in contrast to 5% of nonrunaways. In a logistic regression model, childhood neglect increases the risk of sexual assault between Waves 1 and 2 by nearly two times. Poor mental health is statistically significant. Alcohol use doubles the odds of sexual assault. The risk of sexual assault is approximately three-fold for girls with a history of sexual onset and sexual touching in a romantic relationship. Running away increases the risk by nearly two and a half times. There is evidence that alcohol use and sexual onset partially mediates the relationship between running away and sexual assault.

[Co-localization analysis between porcine epidemic diarrhea virus nucleocapsid protein and nucleolar phosphoprotein B23.1].

OBJECTIVE: To elucidate the co-localization characteristic between porcine epidemic diarrhea virus (PEDV) N protein and B23.1 phosphoprotein. METHODS: Two pairs of primers used to amplify N gene and B23.1 gene were designed and synthesized according to CV777 N gene sequence (AF353511) and human nucleolar phosphoprotein B23.1 gene sequence (BC050628.1), respectively. The PEDV N gene and B23.1 gene were amplified by RT-PCR from PEDV strain CV777 and Vero E6 cells, respectively; then cloned into eukaryotic expression vector pAcGFP1-C1 and pDsRed2-N1, to generate the recombinant plasmids pAcGFP1-C1/N and pDsRed2-N1/B23.1, respectively. Vero E6 cells were transfected with plasmids pAcGFP1-C1/N and pDsRed2-N1/B23.1. RESULTS: The fusion proteins successfully expressed in transfected Vero E6 cells by western blot analysis, and the PEDV N protein and the B23.1 phosphoprotein showed co-localization features in co-transfected cells through confocal microscopy analysis. CONCLUSION: The results will help to identify the nucleolar localization signals in PEDV N protein and to elucidate the mechanism of N protein located in nucleus.