Predicting adult height from DNA variants in a European-Asian admixed population.

Accurate genomic profiling for adult height is of high practical relevance in forensics genetics. Adult height is a classical reference trait in the field of human complex trait genetics characterized by highly polygenic nature and relatively high heritability. A meta-analysis of genome-wide association studies by the Genetic Investigation of Anthropocentric Traits (GIANT) consortium has identified 697 DNA variants associated with adult height in Europeans; however, whether these variants will still be informative in non-Europeans is still in question. The present study investigated the predictive power of these 697 height-associated SNPs in 687 Uyghurs of European-Asian admixed origin. Among all GIANT SNPs, 11% showed nominally significant association (6.78 × 10-4 < p < 0.05) with adult height in the Uyghur population and among the significant SNPs 77% of allele effects were in the same direction as those in Europeans reported in the GIANT study. Fitting linear and logistic models using a polygenic score consisting of all GIANT SNPs resulted in an 80-20 cross-validated mean R2 of 10.08% (95% CI 3.16-18.40%) for quantitative height prediction and a mean AUC value of 0.65 (95% CI 0.57-0.72%) for qualitative "above average" prediction. Fine-tuning the SNP set using their association p values considerably improved the prediction results (number of SNPs = 62, R2 = 15.59%, 95% CI 6.80-25.71%; AUC = 0.70, 95% CI 62-0.77) in the Uyghurs. Overall, our findings demonstrate substantial differences between the European and Asian populations in the genetics of adult height, emphasizing the importance of population heterogeneity underlying the genetic architecture of adult height.

[Study of reconstruction of digital three-dimensional model of normal human placental vascular network based on MRI data in vitro].

OBJECTIVE: To investigate the reconstruction of digital three-dimensional (3D) model of normal human placental vascular network based on MRI data in vitro. METHODS: Six full term placentas were collected, casted with modified self-curing denture base resin and scanned by T1 e-THRIVE high resolution magnetic resonance imaging. MRI images were imported into Mimics 14.0 software for 3D reconstruction, and the 3D model was compared with placental vascular casting model. RESULTS: (1) The placental vascular network could be obtained on MR 2D images. The 3D model were reconstructed successfully, which showed clear, realistic images. The 3D model could be zoomed and revolved from any direction to observe the branches of arteries and veins. (2) The umbilical vein and 2 umbilical arteries could be seen in the 3D model. In the root of the umbilical cord, the umbilical vein divided into 5-7 branches. While the 2 umbilical arteries anatomoses to form blood sinus and then devided into sub-branches. All the peripheral vessels ended in chorionic plate with abundant sub-branches. (3) When compared with the casting of placental arterial-venous vascular network, the morphology, structure, angle and trend of vessels in 3D model was consistent with the casting network. CONCLUSIONS: Reconstruction of digital 3D model of normal human placental vascular network based on MRI in vitro is a new and promising method for the study of placental vasculature. It has better vascular exposure, free rotation, radiation-free. It provides a promising base for the study of placental vasculature in vivo in the future.

Studies of bicalutamide-excipients interaction by combination of molecular docking and molecular dynamics simulation.

While the effects of hydrophilic excipients in enhancing the dissolution rate of water-insoluble drugs have been validated, the underlying mechanism remains poorly understood, particularly at a molecular level. In this work, a combination of docking calculations and MD simulations was applied to investigate the molecular interactions between bicalutamide (BIC) and each of three excipients: lactose (LAC), hydroxypropyl methylcellulose (HPMC), and mannitol (MAN). The calculated results indicated that BIC interacted with HPMC and MAN mainly by Lennard-Jones (LJ) interactions but with LAC mainly by Coulomb (Coul) interactions. There was no hydrogen bond formed between BIC and excipient. It was shown that BIC/LAC had the biggest total solvent accessible surface area with the biggest hydrophilic area and formed the most hydrogen bonds between excipient and water. In addition to the structure analyses, BIC/LAC had both the lowest interaction energy between BIC and excipient and the lowest interaction energy between BIC/excipient and water. All these led to the best dissolution performance of BIC/LAC, which could correspond to the experimental results of dissolution test. The present study suggests that a combination of docking calculations and MD simulations, which aims at complementing the experimental work, could provide a molecular insight into the interaction between drug and excipient. It also holds the great potential to simplify the optimization process of drug delivery system and reduce both time and costs.

[Characteristics and Risk Evaluation of Heavy Metal Contamination in Paddy Soils in the Three Gorges Reservoir Area].

Soil Cd, Hg, Pb, As, Cr, Cu, Zn, and Ni of 12 districts in the Three Gorges Reservoir area (Chongqing section) were analyzed, and different evaluation methods were used to assess the degree of contamination, potential ecological risk, and human health risk of soil heavy metals in paddy soils. The results showed that the average values of all heavy metals except Cr in paddy soils in the Three Gorges Reservoir area exceeded the background values of soils in the Three Gorges Reservoir area, and the contents of Cd, Cu, and Ni in 12.32%, 4.35%, and 2.54% of the soil samples exceeded the screening values, respectively. The variation coefficients of the eight heavy metals were 29.08%-56.43%, which belonged to the medium and above-intensity variation levels and were influenced by anthropogenic activities. The eight heavy metals were contaminated in the soil, and 16.30%, 6.52%, and 2.90% of the soil Cd, Hg, and Pb were heavily contaminated. At the same time, the potential ecological risk of soil Hg and Cd were in the medium risk level on the whole. Wuxi County and Wushan County had relatively high pollution levels among the 12 districts, the Nemerow pollution index showed a moderate pollution level, and the comprehensive potential ecological risks were also at a moderate ecological hazard level. The results of the health risk evaluation showed that hand-mouth intake was the main exposure path of non-carcinogenic risk and carcinogenic risk. Soil heavy metals presented no non-carcinogenic risk for adults (HI<1), but 12.68% of the sites had non-carcinogenic risk for children (HI>1). As and Cr were the main influencing factors for non-carcinogenic and carcinogenic risks in the study area, and their total contributions to non-carcinogenic and carcinogenic risks were more than 75% and 95%, respectively, which was cause for concern.

Synergistic effect of poly(ionic liquid) and phosphoramide on flame retardancy and crystallization of poly(lactic acid).

Crystallinity and flame retardancy are two key properties for poly(lactic acid)(PLA) in applications. In this paper, a quaternary phosphonium salt poly(ionic liquid) (PIL) and a phosphamide (POFA) were prepared. The PIL, POFA and their blend were used to regulate the flame retardancy and crystallization behaviors of PLA using the limiting oxygen index, UL-94 vertical burning, and thermogravimetric analysis, and differential scanning calorimetry etc. The results showed that a synergistic effect exists between PIL and POFA on flame retardancy. When 6 wt% PIL/POFA (2/1) was added into PLA, its LOI value is 28.0 vol%, and achieves the UL-94 V-0 rating while the PLA composites containing 6 wt% PIL or POFA just achieve the UL-94 V2. The PIL/POFA improves the flame retardancy of PLA by melting-away mode. In addition, the crystallization rate of PLA containing PIL/POFA is faster than that of PLA/PIL and PLA/POFA. The degradation of PLA induced by PIL/POFA produces some small molecular oligomers, which enhances the molecular chain mobility and rearrangement, thus contributes to better flame retardancy and faster crystallization.

Association study of monoamine oxidase A/B genes and schizophrenia in Han Chinese.

BACKGROUND: Monoamine oxidases (MAOs) catalyze the metabolism of dopaminergic neurotransmitters. Polymorphisms of isoforms MAOA and MAOB have been implicated in the etiology of mental disorders such as schizophrenia. Association studies detected these polymorphisms in several populations, however the data have not been conclusive to date. Here, we investigated the association of MAOA and MAOB polymorphisms with schizophrenia in a Han Chinese population. METHODS: Two functional single nucleotide polymorphisms (SNPs), rs6323 of MAOA and rs1799836 of MAOB, were selected for association analysis in 537 unrelated schizophrenia patients and 536 healthy controls. Single-locus and Haplotype associations were calculated. RESULTS: No differences were found in the allelic distribution of rs6323. The G allele of rs1799836 was identified as a risk factor in the development of schizophrenia (P = 0.00001). The risk haplotype rs6323T-rs1799836G was associated with schizophrenia in female patients (P = 0.0002), but the frequency difference was not significant among male groups. CONCLUSIONS: Our results suggest that MAOB is a susceptibility gene for schizophrenia. In contrast, no significant associations were observed for the MAOA functional polymorphism with schizophrenia in Han Chinese. These data support further investigation of the role of MAO genes in schizophrenia.

Emergence of a non vaccine-cognate enterovirus A71 genotype C1 in mainland China.

BACKGROUND: EV-A71 is a common causative agent of hand foot and mouth disease. In mainland China, EV-A71 subgenotype C4 has been the sole circulating genotype since 2008, and was used in the production of multiple licensed vaccines. Here, we report the first detection EV-A71 C1 strains in China. METHODS: Full genomic sequence were obtained. The origin of the EV-A71 C1 strains were tracked down by Bayesian inferences. Recombination was analyzed using Simplot program. And the antigenicity were tested using the microneutralization test. RESULTS: The C1-GD2019 shared high identity with the C1-like lineage recently identified in Europe and was introduced into Guangdong in 2018-2019. Close genetic relatedness between the C1-GD2019 and Europe C1-like strains were observed except for the 3D-3'UTR region. The late showed high similarity with CVA genomes. Antigenic variance was found. The C1-GD2019 could not be effectively neutralized by EV-A71 C4a neutralizing antibody positive samples. CONCLUSION: This is the first report of EV-A71 subgenotype C1 isolated in China. It is a recombinant strain originating from C1-like strains recently identified in Europe and CVA strains. The different antigenicity between the C1 strains and C4a vaccine strains highlighted the importance on closely monitoring the EV-A71 C1 strains in China.

Design, preparation and characterization of cyclic RGDfK peptide modified poly(ethylene glycol)-block-poly(lactic acid) micelle for targeted delivery.

Molecular targeted cancer therapy is a promising strategy to overcome the lack of specificity of anticancer drug. While the binding of c(RGDfK) (cyclic Arginine-Glycine-Aspartic acid-Phenylalanine-Lysine) to αvß3 over-expressed on tumor cell has been validated, the underlying interaction remains poorly understood. In this work, docking calculation was applied to investigate the interactions between c(RGDfK)/c(RGDfK)-PEG and αvß3. The calculated results indicated that c(RGDfK) interacted with αvß3 mainly by electrostatic interaction, stabilization interaction, and hydrophobic interaction. Conjugation of PEG chain to the c(RGDfK) weakened the binding affinity of c(RGDfK) to αvß3. Accordingly, docetaxel(DTX)-loaded target micelles (c(RGDfK)-PEG-PLA/PEG-PLA/DTX) were designed, characterized and evaluated using HeLa cells. In vitro release studies demonstrated both target and non-target micelles displayed almost the same profiles, which best fit in Ritger-Peppas model. Cellular uptake and MTT studies revealed that the target micelles with the presence of c(RGDfK) were more efficiently taken up by HeLa cells and significantly improved the cytotoxicity compared to that of non-target micelles. Cell inhibition rate of target micelles was improved by 20% after 24h. Our findings suggest that target micelles may be a potential anticancer drug delivery system in the treatment of integrin αvß3 over-expressed on tumor cell.

Generation of Tg(cyp1a:gfp) Transgenic Zebrafish for Development of a Convenient and Sensitive In Vivo Assay for Aryl Hydrocarbon Receptor Activity.

Both dioxins/dioxin-like compounds and polycyclic aromatic hydrocarbons (PAHs) are persistent organic pollutants and cause multiple adverse health effects on human and wildlife. Cyp1a is the most commonly used biomarker induced by these pollutants through activation of the aryl hydrocarbon receptor (AhR) pathway. Here we generated Tg(cyp1a:gfp) transgenic zebrafish for establishing a convenient in vivo assay for analysing these xenobiotic compounds. The Tg(cyp1a:gfp) larvae at 4 day post-fertilization were tested with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and GFP induction was observed mainly in the kidney, liver and gut. Similar GFP expression was also induced strongly by two dioxin-like chemicals, co-planar polychlorinated biphenyl (PCB126) and polychlorinated dibenzo-p-furan (PeCDF) and relatively weakly by two PAHs, 3-methylcholanthrene (3-MC) and benzo[a]pyrene (BAP). The lowest observed effective concentration (LOEC) of TCDD was estimated to be ∼1 pM and the EC50 (effective concentration to induce GFP in 50 % of Tg(cyp1a:gfp) larvae) was ∼10 pM. PCB126 and PeCDF had ∼10× lower potencies in GFP induction than TCDD, while the potencies for 3-MC and BAP were at least 1000× lower. The sensitivity of Tg(cyp1a:gfp) larvae to respond TCDD was also favourable compared to that of ethoxyresorufin-O-deethylase (EROD) assay in both zebrafish larvae and adult livers. As GFP-based assay in transgenic zebrafish can be easily accommodated in multi-well dishes, the Tg(cyp1a:gfp) zebrafish should provide not only a valuable biomonitoring tool for aquatic contaminants but also a potential high-throughput chemical screening platform for identification of new AhR agonists.

An efficient method for in vitro gene delivery via regulation of cellular endocytosis pathway.

Transfection efficiency was the primary goal for in vitro gene delivery mediated by nonviral gene carriers. Here, we report a modified gene transfection method that could greatly increase the efficiency of, and accelerate the process mediated by, 25 kDa branched polyethyleneimine and Lipofectamine™ 2000 in a broad range of cell strains, including tumor, normal, primary, and embryonic stem cells. In this method, the combination of transfection procedure with optimized complexation volume had a determinant effect on gene delivery result. The superiorities of the method were found to be related to the change of cellular endocytosis pathway and decrease of particle size. The efficient and simple method established in this study can be widely used for in vitro gene delivery into cultured cells. We think it may also be applicable for many more nonviral gene delivery materials than polyethyleneimine and liposome.

Pluronic L64-mediated stable HIF-1α expression in muscle for therapeutic angiogenesis in mouse hindlimb ischemia.

Intramuscular injection of plasmid DNA (pDNA) to express a therapeutic protein is a promising method for the treatment of many diseases. However, the therapeutic applications are usually hindered by gene delivery efficiency and expression level. In this study, critical factors in a pDNA-based gene therapy system, such as gene delivery materials, a therapeutic gene, and its regulatory elements, were optimized to establish an integrated system for the treatment of mouse hindlimb ischemia. The results showed that Pluronic L64 (L64) was an efficient and safe material for gene delivery into mouse skeletal muscle. It also showed intrinsic ability to promote in vivo angiogenesis in a concentration-dependent manner, which might be through the activation of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB)-regulated angiogenic factors. The combination of 0.1% L64 with a hybrid gene promoter (pSC) increased the gene expression level, elongated the gene expression duration, and enhanced the number of transfected muscle fibers. In mice ischemic limbs, a gene medicine (pSC-HIF1α(tri)/L64) composed of L64 and pSC-based expression plasmid encoding hypoxia-inducible factor 1-alpha triple mutant (HIF-1α(tri)), improved the expression of stable HIF-1α, and in turn, the expression of multiple angiogenic factors. As a result, the ischemic limbs showed accelerated function recovery, reduced foot necrosis, faster blood reperfusion, and higher capillary density. These results indicated that the pSC-HIF1α(tri)/L64 combination presented a potential and convenient venue for the treatment of peripheral vascular diseases, especially critical limb ischemia.

A filter paper-based microdevice for low-cost, rapid, and automated DNA extraction and amplification from diverse sample types.

A plastic microfluidic device that integrates a filter disc as a DNA capture phase was successfully developed for low-cost, rapid and automated DNA extraction and PCR amplification from various raw samples. The microdevice was constructed by sandwiching a piece of Fusion 5 filter, as well as a PDMS (polydimethylsiloxane) membrane, between two PMMA (poly(methyl methacrylate)) layers. An automated DNA extraction from 1 µL of human whole blood can be finished on the chip in 7 minutes by sequentially aspirating NaOH, HCl, and water through the filter. The filter disc containing extracted DNA was then taken out directly for PCR. On-chip DNA purification from 0.25-1 µL of human whole blood yielded 8.1-21.8 ng of DNA, higher than those obtained using QIAamp® DNA Micro kits. To realize DNA extraction from raw samples, an additional sample loading chamber containing a filter net with an 80 µm mesh size was designed in front of the extraction chamber to accommodate sample materials. Real-world samples, including whole blood, dried blood stains on Whatman® 903 paper, dried blood stains on FTA™ cards, buccal swabs, saliva, and cigarette butts, can all be processed in the system in 8 minutes. In addition, multiplex amplification of 15 STR (short tandem repeat) loci and Sanger-based DNA sequencing of the 520 bp GJB2 gene were accomplished from the filters that contained extracted DNA from blood. To further prove the feasibility of integrating this extraction method with downstream analyses, "in situ" PCR amplifications were successfully performed in the DNA extraction chamber following DNA purification from blood and blood stains without DNA elution. Using a modified protocol to bond the PDMS and PMMA, our plastic PDMS devices withstood the PCR process without any leakage. This study represents a significant step towards the practical application of on-chip DNA extraction methods, as well as the development of fully integrated genetic analytical systems.

Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.

BACKGROUND: Effective gene transfection without serum deprivation is a prerequisite for successful stem cell-based gene therapy. Polyethylenimine (PEI) is an efficient nonviral gene vector, but its application has been hindered by serum sensitivity and severe cytotoxicity. METHODS: To solve this problem, a new family of lipopolyplexes was developed by coating PEI/DNA polyplexes with three serum-resistant cationic lipids, namely, lysinylated, histidylated, and arginylated cholesterol. The physical properties, transfection efficiency, cellular uptake, subcellular distribution, and cytotoxicity of the lipopolyplexes was investigated. RESULTS: The outer coat composed of lysinylated or histidylated cholesterol remarkably improved the transfection efficiency of the polyplex with a low PEI/DNA ratio of 2 in the presence of serum. The resulting lysinylated and histidylated cholesterol lipopolyplexes were even more efficient than the best performing polyplex with a high PEI/DNA ratio of 10. Results from cellular uptake and subcellular distribution studies suggest that their higher transfection efficiency may result from accelerated DNA nuclear localization. The superiority of the lipopolyplexes over the best performing polyplex was also confirmed by delivering the therapeutic gene, hVEGF(165). Equally importantly, the lipid coating removed the necessity of introducing excess free PEI chains into the transfection solution for higher efficiency, generating lipopolyplexes with no signs of cytotoxicity. CONCLUSION: Noncovalent modification of polyplexes with lysinylated and histidylated cholesterol lipids can simultaneously improve efficiency and reduce the toxicity of gene delivery under serum conditions, showing great promise for genetic modification of bone marrow stem cells.

Transcatheter arterial chemoembolization versus systemic methotrexate for the management of cesarean scar pregnancy.

OBJECTIVE: To evaluate the effectiveness/safety of systemic methotrexate (MTX) treatment versus transcatheter arterial chemoembolization using different embolic agents for termination of cesarean scar pregnancy (CSP). METHODS: Women with CSP were randomized to receive intravenous infusion of MTX (group 1, n=13), or chemoembolization with MTX and either gelatin sponge (GS; group 2, n=15) or polyvinyl alcohol (PVA; group 3, n=16) particles. Uterine suction curettage followed all procedures. Bleeding volume, time until resolution of serum ß-hCG, and length of hospital stay were recorded as outcome endpoints. RESULTS: Bleeding volume was smaller in groups 2 (mean ± SD, 73±20 mL) and 3 (63±22 mL) than in group 1 (952±471 mL) (P<0.001). Time until resolution of ß-hCG was shorter in groups 2 (29±16 days) and 3 (30±19 days) than in group 1 (57±25 days) (P<0.01). Length of hospital stay was shorter in groups 2 (13±4 days) and 3 (12±3 days) than in group 1 (36±8 days) (P<0.01). CONCLUSION: Transcatheter arterial chemoembolization was more effective than systemic MTX treatment for termination of CSP. Large cohort studies are warranted to compare effectiveness between PVA and GS particles.