Preparation of ß-CD-Vitexin Microspheres and their Effects on SW480 Cell Proliferation.

OBJECTIVE: In order to overcome the insolution and low bioavailability of the vitexin in vivo, ß-cyclodextrin-vitexin (ß-CD-vitexin) microspheres were prepared, and their effects on the proliferation of SW480 cells were observed. METHODS: Scanning electron microscopy, ultraviolet spectrum, Fourier transform infrared spectroscopy, and release rate analysis identified the formation of ß-CD-vitexin microspheres. MTT assay detected the effect of ß-CD-vitexin microspheres on tumor cell proliferation at 6, 12, 24, and 48 h. Fluorescence microscopy and flow cytometry were used to observe the effect of ß-CD-vitexin microspheres on the apoptosis of SW480 cells. The mRNA expression of the p53 gene was measured by qPCR. RESULTS: ß-CD-vitexin microspheres were successfully prepared. SW480 cell proliferation was inhibited by 0.1, 0.2, and 0.4 mg/mL of ß-CD-vitexin microspheres in a dose- and time-dependent manner, and the mechanism of proliferation inhibition was related to cell apoptosis caused by the upregulated expression of p53 gene. CONCLUSION: The preparation of ß-CD-vitexin sustained release microspheres is feasible, and ß-CDvitexin microspheres have potential anti-colorectal cancer value.

Distribution and model prediction of antibiotic resistance genes in Weishan Lake based on the indication of Chironomidae larvae.

The widespread contamination of antibiotic resistance genes (ARGs) in freshwater environment are becoming a serious challenge to human health and ecological safety. Rapid and efficient monitoring of ARGs pollution is of great significance to ARGs control. Water, bottom mud, and fish have all been used to indicate ARG contamination in aquatic environments. However, it is unclear whether macrobenthic invertebrates in the food chain of aquatic environments can be indicators of ARG contamination. In this study, we demonstrated that ARGs including tetA gene, sul2 gene, and km gene were distributed in Chironomidae larvae in Weishan Lake. The ARG distribution was related to animal species, body parts, sampling sites, time, urban environment, animal farming, south-to-north water diversion, food chain, antibiotics, and water storage. Mathematical model predictions of ARG contamination in Weishan Lake were constructed based on the structural equation model (SEM) and the distribution of ARG sul2 in Chironomidae larvae. Influencing factors such as water storage, metal elements, antibiotic, and temperature were found to be closely related to the prediction of ARG contamination. This study provided a new indicator for ARG contamination in freshwater environments and a method to predict ARGs contamination.

Protective effect of hawthorn vitexin on the ethanol-injured DNA of BRL-3A hepatocytes.

ABSTRACT: Vitexin is a natural active ingredient in hawthorn leaves, which has a wide range of anti-tumor effects. This study was conducted to assess the protective effect of hawthorn vitexin on the ethanol-injured DNA of hepatocytes in vitro and to explore its mechanism. The effect of different concentrations of hawthorn vitexin on ethanol-injured hepatocytes was detected via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method to study the protective effect of hawthorn vitexin on ethanol-injured DNA damage in hepatocytes. Single-cell gel electrophoresis was used to observe the effect of hawthorn vitexin on ethanol-induced DNA damage in hepatocytes, and the Olive tail moment was measured. Cell physiological and biochemical indexes, such as superoxide dismutase activity, malonaldehyde content, and glutathione peroxidase activity, were detected with kits. The mRNA expression of the superoxide dismutase gene was measured via real-time quantitative polymerase chain reaction. It was showed that 0.2, 0.4, and 0.8 mg mL-1 hawthorn vitexin could significantly repair hepatocyte growth and ethanol-induced DNA damage. This effect was closely related to the improvement in superoxide dismutase, malonaldehyde, and glutathione peroxidase. Hawthorn vitexin could be used to repair ethanol-injured hepatocytes through antioxidation effects, and showed potential for the treatment of liver injury.

Nano-Al2O3 can mediate transduction-like transformation of antibiotic resistance genes in water.

The prevalence of various antibiotic resistance genes (ARGs) and resistant bacteria has caused global public health risks. The carrier transport mediated by phages or membrane vesicles is an important way for horizontal transfer of ARGs. Nano metal oxide particles (NMOPs), which can enter cell through the cell membrane, may be used as the carriers of genes. However, whether they can be used as transmembrane delivery vectors for the horizontal ARG transfer remains unknown. Here, we set up a model of MONPs-mediated transfer of ARGs, and demonstrate that NMOPs, especially for nano-Al2O3, can act as carriers mediating the transduction-like ARG transformation in water. The highest transfer rate mediated by nano-Al2O3 is 4.53 × 104 cfu/mmol, and it is 104 times higher than that of control. Nano-Al2O3 can combine with plasmid coding for ARGs to form high-density package and prevent ARGs from degradation by endonuclease. The results of superresolution fluorescence microscopy and transmission electron microscopy show that nano-Al2O3 can carry ARGs for transmembrane transport. Genome-wide transcription microarray and qPCR indicate that SOS response was closely related to transduction-like ARG transformation mediated by nano-Al2O3. This study is the first to demonstrate that as a new transmembrane carrier, nano-Al2O3 can also cause ARGs diffusion in water.

Detecting Genetic Ancestry and Adaptation in the Taiwanese Han People.

The Taiwanese people are composed of diverse indigenous populations and the Taiwanese Han. About 95% of the Taiwanese identify themselves as Taiwanese Han, but this may not be a homogeneous population because they migrated to the island from various regions of continental East Asia over a period of 400 years. Little is known about the underlying patterns of genetic ancestry, population admixture, and evolutionary adaptation in the Taiwanese Han people. Here, we analyzed the whole-genome single-nucleotide polymorphism genotyping data from 14,401 individuals of Taiwanese Han collected by the Taiwan Biobank and the whole-genome sequencing data for a subset of 772 people. We detected four major genetic ancestries with distinct geographic distributions (i.e., Northern, Southeastern, Japonic, and Island Southeast Asian ancestries) and signatures of population mixture contributing to the genomes of Taiwanese Han. We further scanned for signatures of positive natural selection that caused unusually long-range haplotypes and elevations of hitchhiked variants. As a result, we identified 16 candidate loci in which selection signals can be unambiguously localized at five single genes: CTNNA2, LRP1B, CSNK1G3, ASTN2, and NEO1. Statistical associations were examined in 16 metabolic-related traits to further elucidate the functional effects of each candidate gene. All five genes appear to have pleiotropic connections to various types of disease susceptibility and significant associations with at least one metabolic-related trait. Together, our results provide critical insights for understanding the evolutionary history and adaption of the Taiwanese Han population.

Effects of free antibiotic resistance genes in the environment on intestinal microecology of mice.

The rapid spread of antibiotic resistance genes (ARGs) is a great challenge to the ecological safety and human health. The intestine of humans and animals is an important site for the increase and spread of ARGs due to the great diversity and abundance of microorganisms in the intestinal microecology. ARGs, including the intracellular (iARGs) and the extracellular (eARGs) ARGs, are usually introduced into the intestinal tract through the diet, and the iARGs are colonized and spread in the intestinal microbiota with the help of the host bacteria. However, whether the eARGs can enter the intestinal microorganisms in the absence of host bacteria is not known. Here, we show the transformation and the diffusion of the ampramycin resistance gene (Ap) carried by the free plasmid RK2 in the intestinal microbiota of mice. After two days of consecutive gavage with free RK2, the intracellular Ap gene increases from days 0-8 in the feces of mice, and has remained constant. Bacterial transformation happens in the small intestine, including proximal and distal jejuna and proximal and distal ilea, at the early stage (first two days), and the intracellular RK2 is diffused into the intestinal microbiota of mice by conjugation on days 2-8 day, which is based on the distribution of eARG and iARG and the mRNA expression levels of trbBp, trfAp, korA, korB, and trbA. The characteristics of ARGs susceptible microbiota for transformation are analyzed using 16s rRNA gene sequencing, transmission electron microscopy, and flow cytometric. The ingestion of RK2 affects the composition of intestinal microbiota especially for Proteobacteria, and the antibiotic residue promotes the increase in Escherichia coli. These findings are important to assess the risk of ARGs, especially the eARGs in the intestinal microecology.

Preparation of ß-CD-Quercetin Complex and its Effects on Ethanol- Damaged BRL-3A Hepatocytes.

OBJECTIVE: To prepare the sustained-release complex, quercetin was incorporated with ß- cyclodextrin (ß-CD) and the effect of ß-CD-quercetin complex on the growth of ethanol-injuried hepatocytes was studied. METHODS: By using scanning electron microscopy, infrared spectroscopy, and release rate analysis, ß- CD-quercetin complex was identified. The effect of different concentrations of ß-CD-quercetin complex on the growth of ethanol-damaged hepatocytes at different time was observed by using MTT assay, and the cell quantity and morphology were observed by using hematoxylin-eosin staining. By using single-cell gel electrophoresis, the prevention of ß-CD-quercetin complex from the DNA damage of ethanol-damaged BRL-3A cells was studied, and Olive tail moment was calculated. RESULTS: ß-CD-quercetin complex as the sustained-release complex was successfully prepared. The ethanol induced damage of BRL-3A cells could be prevented by 20, 40 and 80 mg/L of quercetin complex, and the protection mechanism of hepatocyte was related to the antioxidation of DNA. CONCLUSION: Quercetin sustained-release complex could be prepared with ß-CD, and it might be used to treat alcoholic liver disease.

Preparation of Chitosan/Alginate-ellagic Acid Sustained-release Microspheres and their Inhibition of Preadipocyte Adipogenic Differentiation.

OBJECTIVE: In this study, chitosan/alginate-ellagic acid sustained-release microspheres were prepared, and the effect of sustained-release microspheres on preadipocyte adipogenic differentiation was analyzed. METHODS: Chitosan/alginate-ellagic acid microspheres were prepared and identified by scanning electron microscopy (SEM) and infrared spectroscopy (IR). The drug release rates were measured at pH 6.8, 7.0, 7.2, 7.4 to determine sustained release of ellagic acid from microspheres. The effects of 0.1, 1, 10 mg/L chitosan/alginate-ellagic acid microsphere on 3T3-F442A preadipocyte proliferation were determined by Methyl thiazolyl tetrazolium assay (MTT), and cell morphology was checked by hematoxylin/ eosin staining (HE staining). The effect of chitosan/alginate-ellagic acid microspheres on preadipocyte adipogenic differentiation was also determined by Oil red O staining, and lipogenesis was measured by isopropanol extraction. The molecular mechanism was investigated by detecting the mRNA expression of CCAAT/enhancer binding protein alpha (C/EBPα) and peroxisome proliferatorsactivated receptor gamma (PPARγ). RESULTS: Chitosan/alginate-ellagic acid sustained-release microspheres were successfully prepared, and the inhibition of proliferation and adipogenic differentiation of preadipocytes was found to be dosedependent. The mechanism of differentiation inhibition was found to be closely related to the expression of transcription factor C/EBPα and PPARγ. CONCLUSION: Chitosan/alginate can be used as a good material to prepare ellagic acid sustained-release microspheres, and these microspheres can be used for treating the obesity.

Rational Design and General Synthesis of S-Doped Hard Carbon with Tunable Doping Sites toward Excellent Na-Ion Storage Performance.

Heteroatom-doping is a promising strategy to tuning the microstructure of carbon material toward improved electrochemical storage performance. However, it is a big challenge to control the doping sites for heteroatom-doping and the rational design of doping is urgently needed. Herein, S doping sites and the influence of interlayer spacing for two kinds of hard carbon, perfect structure and vacancy defect structure, are explored by the first-principles method. S prefers doping in the interlayer for the former with interlayer distance of 3.997 Å, while S is doped on the carbon layer for the latter with interlayer distance of 3.695 Å. More importantly, one step molten salts method is developed as a universal synthetic strategy to fabricate hard carbon with tunable microstructure. It is demonstrated by the experimental results that S-doping hard carbon with fewer pores exhibits a larger interlayer spacing than that of porous carbon, agreeing well with the theoretical prediction. Furthermore, the S-doping carbon with larger interlayer distance and fewer pores exhibits remarkably large reversible capacity, excellent rate performance, and long-term cycling stability for Na-ion storage. A stable and reversible capacity of ≈200 mAh g-1 is steadily kept even after 4000 cycles at 1 A g-1 .

Self-defense of Escherichia coli against damages caused by nanoalumina.

Although studies showed effects of nanoalumina (nano-Al2O3) on Escherichia coli, no study completely provides understanding on how bacterial cells respond to damages, especially on how they initiate self-defense. In this study, we showed three types of responses of E. coli to damages caused by nano-Al2O3. Live, dead, and injured, bacteria showed improved survival rates reaching 104%, 116%, and 104% after exposure to 0.1, 1, and 10mmol/L of nano-Al2O3 respectively. Survival rates improved from 100% to 114%, corresponding to an exposure time of 0-9h, and from 100% to 127%, corresponding to 0-1000µg/L Al3+. Improvements were noted in survival rates of E. coli K12 MG1655, HB101, DH5α, and K12 MG1655 △lexA treated by nano-Al2O3 in Luria-Bertani (LB) exposure system or K12 MG1655 in LB, normal saline(NS) and H2O exposure system. Bacterial cells transformed from long rods to ellipsoidal or nearly spherical as form of self-preservation mechanism; this phenomenon may be related to changes in membrane potential induced by free Al3+ released from nano-Al2O3 particles. Molecular mechanism of this response involved inhibited gene expression of sythesis and metabolism of carbohydrates, lipids and proteins. Findings presented in this study may improve understanding of potential danger of nanomaterials and control their spread to environmen.

Synthesis of Mesoporous Co2+-Doped TiO2 Nanodisks Derived from Metal Organic Frameworks with Improved Sodium Storage Performance.

TiO2 is a most promising anode candidate for rechargeable Na-ion batteries (NIBs) because of its appropriate working voltage, low cost, and superior structural stability during chage/discharge process. Nevertheless, it suffers from intrinsically low electrical conductivity. Herein, we report an in situ synthesis of Co2+-doped TiO2 through the thermal treatment of metal organic frameworks precursors of MIL-125(Ti)-Co as a superior anode material for NIBs. The Co2+-doped TiO2 possesses uniform nanodisk morphology, a large surface area and mesoporous structure with narrow pore distribution. The reversible capacity, Coulombic efficiency (CE) and rate capability can be improved by Co2+ doping in mesoporous TiO2 anode. Co2+-doped mesoporous TiO2 nanodisks exhibited a high reversible capacity of 232 mAhg-1 at 0.1 Ag1-, good rate capability and cycling stability with a stable capacity of about 140 mAhg-1 at 0.5 Ag1- after 500 cycles. The enhanced Na-ion storage performance could be due to the increased electrical conductivity revealed by Kelvin probe force microscopy measurements.

Preparation of ß-CD-Ellagic Acid Microspheres and Their Effects on HepG2 Cell Proliferation.

OBJECTIVE: In this study, ß-cyclodextrin (ß-CD) was chosen as the coating for ellagic acid to prepare ellagic acid microspheres, and the effect of microspheres on the growth of HepG2 cells was observed. METHODS: Scanning electron microscopy, infrared spectroscopy, and release rate analysis were used to identify the formation of ellagic acid microspheres. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effect of different concentrations of ellagic acid microspheres on tumor cell proliferation at 6, 12, 24 and 36 h, and cell morphology and quantity were observed using hematoxylin-eosin (HE) staining. Single-cell gel electrophoresis was used to observe the effect of ellagic acid microspheres on the DNA damage of HepG2 cells, and the Olive tail moment and the mRNA expression of tumor suppressor protein gene p53 was measured. RESULTS: ß-CD could be used as wrapping material of ellagic acid to prepare ellagic acid microspheres. HepG2 cell proliferation could be inhibited by 0.1, 0.3 and 0.5 g/L of ellagic acid microspheres in a dose- and time-dependent manner, and the mechanism of proliferation inhibition was related to DNA damage and cell apoptosis. CONCLUSION: Preparing ellagic acid microspheres with ß-CD is feasible, and ellagic acid microspheres have potential therapeutic value (anticancer).

Efficacy of targeted agents in the treatment of elderly patients with advanced non-small-cell lung cancer: a systematic review and meta-analysis.

PURPOSE: The efficacy of targeted agents (TAs) in the treatment of elderly patients with advanced non-small-cell lung cancer (NSCLC) remains controversial. We aimed to assess the efficacy of TAs in the treatment of advanced NSCLC in this setting. MATERIALS AND METHODS: Relevant trials were identified by searching electronic databases and conference meetings. Prospective randomized controlled trials assessing chemotherapies with or without TAs in elderly patients with advanced NSCLC were included. Outcomes of interest included overall survival (OS) and progression-free survival (PFS) in elderly patients with advanced NSCLC. RESULTS: A total of 4,093 elderly patients from 17 randomized controlled trials were included for analysis. The addition of TAs to chemotherapy significantly improved PFS (hazard ratio [HR] 0.85, 95% confidence interval [CI]: 0.75-0.96, P=0.01) when compared to chemotherapy alone. There was also a tendency to improve OS in the combination groups (HR 0.92, 95% CI: 0.85-1.01, P=0.064). Subgroup analysis based on treatment line indicated that TAs plus chemotherapy as first-line chemotherapy in elderly patients with advanced NSCLC significantly improved PFS (HR 0.80, 95% CI: 0.68-0.95, P=0.01) and OS (HR 0.91, 95% CI: 0.83-0.99, P=0.037), while the use of TA-containing regimens as second-line therapy in these patients did not significantly improve PFS (HR 0.91, 95% CI: 0.75-1.10, P=0.33) and OS (HR 1.04, 95% CI: 0.81-1.33, P=0.77) in comparison with chemotherapy alone. No publication bias was detected by Begg's and Egger's tests for OS. CONCLUSION: The findings of this study suggest that the addition of TAs to first-line chemotherapy in elderly patients with advanced NSCLC offers an improved PFS and OS. Further trials are recommended to clearly investigate the efficacy of adding specific TAs to first-line chemotherapy for advanced NSCLC in this setting.

A fully integrated standalone portable cavity ringdown breath acetone analyzer.

Breath analysis is a promising new technique for nonintrusive disease diagnosis and metabolic status monitoring. One challenging issue in using a breath biomarker for potential particular disease screening is to find a quantitative relationship between the concentration of the breath biomarker and clinical diagnostic parameters of the specific disease. In order to address this issue, we need a new instrument that is capable of conducting real-time, online breath analysis with high data throughput, so that a large scale of clinical test (more subjects) can be achieved in a short period of time. In this work, we report a fully integrated, standalone, portable analyzer based on the cavity ringdown spectroscopy technique for near-real time, online breath acetone measurements. The performance of the portable analyzer in measurements of breath acetone was interrogated and validated by using the certificated gas chromatography-mass spectrometry. The results show that this new analyzer is useful for reliable online (online introduction of a breath sample without pre-treatment) breath acetone analysis with high sensitivity (57 ppb) and high data throughput (one data per second). Subsequently, the validated breath analyzer was employed for acetone measurements in 119 human subjects under various situations. The instrument design, packaging, specifications, and future improvements were also described. From an optical ringdown cavity operated by the lab-set electronics reported previously to this fully integrated standalone new instrument, we have enabled a new scientific tool suited for large scales of breath acetone analysis and created an instrument platform that can even be adopted for study of other breath biomarkers by using different lasers and ringdown mirrors covering corresponding spectral fingerprints.

Case report: Enteral nutritional supplement as a likely cause of false-positive galactomannan testing.

The detection of galactomannan (GM) in the serum of in immunocompromised patients is widely used for the early diagnosis of invasive aspergillosis. We report a case of a false-positive GM test presumably caused by the enteral nutritional supplement given to a non-neutropenic patient with intestinal graft-versus-host disease after a hematopoietic stem cell transplant. Clinicians should be alert to the possibility of false-positive GM results in patients on nutritional supplements.

[The noninvasive measurement research of human blood parameters under flow control of time gate].

In order to further improve the noninvasive measurement precision of human blood components and achieve clinical requirements, we propose a new measurement method based on the dynamic dual wavelength time-resolved transmittance measurement, combined with the advanced time gate technology and Laplasse transform to detect human blood components noninvasively in the blocked blood flow conditions. Simulation results show that when p>0, emphasizing the importance of early arriving photons contribution can enhance the detection sensitivity of human body blood parameters.

Polycyclic aromatic hydrocarbons (PAHs) in soils and vegetation near an e-waste recycling site in South China: concentration, distribution, source, and risk assessment.

This study determined the concentrations of PAHs generated from e-waste recycling activities and their potential impacts on soil, vegetation, and human health. The total PAH concentrations in soils and plants ranged from 127 to 10,600 and 199 to 2420 ng/g, respectively. Samples from an e-waste burning site had higher PAH concentrations than samples from adjacent locations. The PAHs in plants varied with plant species and tissue, and Lactuca sativa L. contained the highest PAHs of all the vegetable species. Various land use types showed different PAH concentrations in soils, with vegetable fields showing higher concentrations than paddy fields. Low molecular weight PAHs, such as phenanthrene, were the predominant congeners in soils, whereas high molecular weight PAHs, such as fluoranthene, pyrene, and benzo[a]anthracene, were enriched in plants relative to soils. Dissimilar PAH profiles in soil and the corresponding vegetation indicated that the uptake of PAHs by plants was selective. A source analysis showed that the contamination by PAHs originated primarily from the open burning of e-waste. The total daily intakes of PAHs and carcinogenic PAHs through vegetables at the e-waste dismantling site were estimated to be 279 and 108 ng/kg/d, respectively, indicating that the consumption of vegetables grown near e-waste recycling sites is risky and should be completely avoided.

EMRSA-15 Bacteremia is not Associated with a Worse Outcome Compared with Bacteremia Caused by Multidrug-Resistant MRSA.

EMRSA-15 (ST22-MRSA-IV) is rapidly replacing the endemic ST239 health care-associated methicillin-resistant Staphylococcus aureus clone in Singapore. A one-year single-centre cohort study of inpatients with MRSA bacteremia was performed to determine if bacteremia caused by EMRSA-15 was associated with worse outcomes compared to bacteremia caused by the endemic ST239 strain. Strains were identified by antibiotypes, and subsequent validation was performed on a selected sample of MRSA strains via pulsed-field gel electrophoresis and staphylococcal chromosome cassette mec typing. Two hundred and twenty-eight patients with MRSA bacteremia were studied; Seventy-three were infected with EMRSA-15. EMRSA-15 and ST239-infected patients were similar regarding gender, frequencies of most co-morbidities, and risk factors for adverse outcomes. Similar numbers of EMRSA-15-infected and ST239-infected patients died (24.7% vs 27.1%, P=0.70) or developed complicated infections (41.1% vs 40.0%, P=0.88). After multivariate analysis, EMRSA-15 as a cause of bacteremia was not significantly associated with either death or development of complicated infections, although inappropriate therapy (5.45-fold, P<0.01) and a respiratory source of bacteremia (4.69, P<0.01) were independently associated with subsequent mortality. The increased propensity of EMRSA-15 for dissemination was not associated with increased virulence in our patients. Further work in determining the mechanisms by which highly transmissible MRSA spreads rapidly is required to better target infection control approaches at these important emerging MRSA clones.

Dissemination of multisusceptible methicillin-resistant Staphylococcus aureus in Singapore.

Analysis of hospital-acquired methicillin-resistant Staphylococcus aureus strains isolated from a tertiary public hospital in Singapore revealed that multisusceptible strains had gradually started to replace the endemic multiresistant strain (ST239-MRSA-III) since 2002. Molecular typing showed that this was a predominantly clonal outbreak of a UK-EMRSA-15 strain (ST22-MRSA-IV).