Automatic high-throughput and non-invasive selection of sperm at the biochemical level.

BACKGROUND: Sperm selection, a key step in assisted reproductive technology (ART), has long been restrained at the preliminary physical level (morphology or motility); however, subsequent fertilization and embryogenesis are complicated biochemical processes. Such an enormous "gap" poses tough problems for couples dealing with infertility, especially patients with severe/total asthenozoospermia . METHODS: We developed a biochemical-level, automatic-screening/separation, smart droplet-TO-hydrogel chip (BLASTO-chip) for sperm selection. The droplet can sense the pH change caused by sperm's respiration products and then transforms into a hydrogel to be selected out. FINDINGS: The BLASTO-chip system can select biochemically active sperm with an accuracy of over 90%, and its selection efficiency can be flexibly tuned by nearly 10-fold. All the substances in the system were proven to be biosafe via evaluating mice fertilization and offspring health. Live sperm down to 1% could be enriched by over 76-fold to 76%. For clinical application to patients with severe/total asthenozoospermia, the BLASTO-chip could select live sperm from human semen samples containing 10% live but 100% immotile sperm. The rates of fertilization, cleavage, early embryos, and blastocysts were drastically elevated from 15% to 70.83%, 10% to 62.5%, 5% to 37.5%, and 0% to 16.67%, respectively. CONCLUSIONS: The BLASTO-chip represents a real biochemical-level technology for sperm selection that is completely independent of sperm's motility. It can be a powerful tool in ART, especially for patients with severe/total asthenozoospermia. FUNDING: This work was funded by the Ministry of Science and Technology of China, the Ministry of Education of China, and the Shenzhen-Hong Kong Hetao Cooperation Zone.

Inferring Causal Relationships Between Metabolites and Polycystic Ovary Syndrome Using Summary Statistics from Genome­Wide Association Studies.

Polycystic ovary syndrome (PCOS) is a disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. Previous studies have suggested that metabolites may play a pivotal mediating role in the progression of phenotypic variations. Although several metabolites had been identified as potential markers for PCOS, the relationship between blood metabolites and PCOS was not comprehensively explored. Previously, Pickrell et al. designed a robust approach to infer evidence of a causal relationship between different phenotypes using independently putative causal SNPs. Our previous paper extended this approach to make it more suitable for cases where only a few independently putative causal SNPs were identified to be significantly associated with the phenotypes (i.e., metabolites). When the most significant SNPs in each independent locus (the independent lead SNPs) with p-values of < 1 × 10-5 were used, 3 metabolites (2-tetradecenoyl carnitine, threitol, 1-docosahexaenoylglycerophosphocholine) causally influencing PCOS and 2 metabolites (asparagine and phenyllactate) influenced by PCOS were identified, (relative likelihood r < 0.01). Under a less stringent threshold of r < 0.05, 7 metabolites (trans-4-hydroxyproline, glutaroyl carnitine, stachydrine, undecanoate, 7-Hoca, N-acetylalanine and 2-hydroxyisobutyrate) were identified. Taken together, this study can provide novel insights into the pathophysiological mechanisms underlying PCOS; whether these metabolites can serve as biomarkers to predict PCOS in clinical practice warrants further investigations.

A de novo mutation of ADAMTS8 in a patient with Wiedemann-Steiner syndrome.

BACKGROUND: Wiedemann-Steiner syndrome (WDSTS) is a rare autosomal dominant disorder caused by mutations in the KMT2A gene and is usually characterized by hairy elbows, short stature, developmental delay, intellectual disability and obvious facial dysmorphism. CASE PRESENTATION: Here, we report a 5-year-old girl with clinical features similar to WDSTS, including postnatal growth delay, retarded intellectual development, and ocular hypertelorism. Through whole-exome sequencing (WES), a frameshift variant of KMT2A was found in the patient but not in her parents' genomic DNA. By bioinformatics analysis, the KMT2A variant was demonstrated to be the top candidate pathogenic variant for the clinical phenotype consistent with WDSTS. Moreover, a duplication of exon 1 in ADAMTS8 (belonging to the zinc metalloproteinase family) was found in the genomic DNA of this patient, which may be responsible for the characteristics that are different from those of WDSTS, including early teething, rapid tooth replacement, and dysplastic enamel. CONCLUSIONS: From the above results, we propose that in our patient, the frameshift variant in KMT2A is the main reason for the WDSTS phenotype, and the unreported mutation in ADAMTS8 may be the candidate reason for other characteristics that are different from those of WDSTS. Therefore, this study not only provides a new KMT2A variant associated with WDSTS but is also a reminder that combined mutations may be present in a case with more characteristics than those seen in WDSTS.

Functional Enrichment Analysis Identifying Regulatory Information Associated with Human Fracture.

Dozens of loci associated with fracture have been identified by genome-wide association studies (GWASs). However, most of these variants are located in the noncoding regions including introns, long terminal repeats, and intergenic regions. Although combining regulation information helps to identify the causal SNPs and interpret the involvement of these variants in the etiology of human fracture, regulation information which was truly associated with fracture was unknown. A novel functional enrichment method GARFIELD (GWAS Analysis of Regulatory of Functional Information Enrichment with LD correction) was applied to identify fracture-associated regulation information, including transcript factor binding sites, expression quantitative trait loci (eQTLs), chromatin states, enhancer, promoter, dyadic, super enhancer and Epigenome marks. Fracture SNPs were significantly enriched in exon (Bonferroni correction, p value < 7.14 × 10-3) at two GWAS p value thresholds through GARFIELD. High level of fold-enrichment was observed in super enhancer of monocyte and the enhancer of chondrocyte (Bonferroni correction, p value < 4.45 × 10-3). eQTLs of 44 tissues/cells and 10 transcription factors (TFs) were identified to be associated with human fracture. These results provide new insight into the etiology of human fracture, which might increase the identification of the causal SNPs through the fine-mapping study combined with functional annotation, as well as polygenic risk score.

Dramatic improvement in the mechanical properties of polydopamine/polyacrylamide hydrogel mediated human amniotic membrane.

Human amniotic membrane (hAM) is a promising material for tissue engineering due to several benefits, including desirable biocompatibility, stem cell source, antibacterial activity, etc. However, because of its low elasticity, the clinical application of hAM is severely restricted. To solve this issue, we employed polydopamine/polyacrylamide (PDA/PAM) hydrogels to toughen hAM. The test results indicated that the PDA/PAM hydrogel can enhance the toughness of hAM dramatically due to the formation of abundant chemical bonds and the strong mechanical properties of the hydrogel itself. Compared to pure hAM, the break elongation and tensile strength of PDA/PAM-toughened hAM rose by 154.15 and 492.31%, respectively. And most importantly, the fracture toughness was almost 15 times higher than untreated hAM. In addition, the cytotoxicity of the PDA/PAM-coated hAM was not detected due to the superior biocompatibility of the chemicals used in the study. Treating hAM with adhesive hydrogels to increase its mechanical characteristics will further promote the application of hAM as a tissue engineering material.

Stigmatization and Interpersonal Deviance Behaviors of Tour Guides: The Influence of Self-Identity Threat and Moral Disengagement.

Severe tour guide stigma is a significant problem hindering tourism development. Based on self-identity threat and moral disengagement theory, this study analyzed the relationship between tour guide stigmatization and tour guides' interpersonal deviance behavior. Survey data collected from 241 tour guides at three different points in time showed that tour guide stigmatization was positively related to tour guides' interpersonal deviance behavior and that self-identity threat mediated this effect. The results also show that moral disengagement moderated the effect of tour guides' self-identity threat on interpersonal deviance behavior, as well as the indirect effect of tour guide stigmatization on tour guides' interpersonal deviance behavior via self-identity threat. This study enriches theoretical research on tour guide stigmatization and offers practical suggestions for solving stigmatization problems for tour guides and organizations.

Identification of Multiple Cracks in Composite Laminated Beams Using Perturbation to Dynamic Equilibrium.

Identification of cracks in beam-type components is significant to ensure the safety of structures. Among the approaches relying on mode shapes, the concept of transverse pseudo-force (TPF) has been well proved for single and multiple crack identification in beams made of isotropic materials; however, there is a noticeable gap between the concept of TPF and its applications in composite laminated beams. To fill this gap, an enhanced TPF approach that relies on perturbation to dynamic equilibrium is proposed for the identification of multiple cracks in composite laminated beams. Starting from the transverse equation of motion, this study formulates the TPF in a composite laminated beam for the identification of multiple cracks. The capability of the approach is numerically verified using the FE method. The applicability of the approach is experimentally validated on a carbon fiber-reinforced polymer laminated beam with three cracks, the mode shapes of which are acquired through non-contact vibration measurement using a scanning laser vibrometer. In particular, a statistic manner is utilized to enable the approach to be feasible to real scenarios in the absence of material and structural information; besides, an integrating scheme is utilized to enable the approach to be capable of identifying cracks even in the vicinity of nodes of mode shapes.

The Use of CRISPR/Cas9 as a Tool to Study Human Infectious Viruses.

Clustered regularly interspaced short palindromic repeats (CRISPR) systems are a set of versatile gene-editing toolkit that perform diverse revolutionary functions in various fields of application such as agricultural practices, food industry, biotechnology, biomedicine, and clinical research. Specially, as a novel antiviral method of choice, CRISPR/Cas9 system has been extensively and effectively exploited to fight against human infectious viruses. Infectious diseases including human immunodeficiency virus (HIV), hepatitis B virus (HBV), human papillomavirus (HPV), and other viruses are still global threats with persistent potential to probably cause pandemics. To facilitate virus removals, the CRISPR/Cas9 system has already been customized to confer new antiviral capabilities into host animals either by modifying host genome or by directly targeting viral inherent factors in the form of DNA. Although several limitations and difficulties still need to be conquered, this technology holds great promises in the treatment of human viral infectious diseases. In this review, we will first present a brief biological feature of CRISPR/Cas9 systems, which includes a description of CRISPR/Cas9 structure and composition; thereafter, we will focus on the investigations and applications that employ CRISPR/Cas9 system to combat several human infectious viruses and discuss challenges and future perspectives of using this new platform in the preclinical and clinical settings as an antiviral strategy.

Measurement and Spatial Variation of Green Total Factor Productivity of the Tourism Industry in China.

Promoting tourism in China using sustainable practices has become a very important issue. In order to analyze temporal characteristics and spatial regularities of green total factor productivity (GTFP), carbon emissions and the consumption of energy related to tourism in China were estimated using a "bottom-up" method. The construction of a measurement framework (including carbon emissions and energy consumption) of GTFP for the tourism industry was also undertaken. The data envelopment analysis (DEA) model and the Malmquist-Luenberger (ML) index were used to measure and calculate tourism GTFP in China between 2007 and 2018, as well as analyze spatio-temporal differences. Results indicate that: (1) carbon emissions and the consumption of energy are increasing, and they have not yet peaked, with traffic associated with tourism accounting for the largest proportion among tourism sectors; the spatial distribution of carbon emissions and the consumption of energy is not balanced; (2) green development of tourism in China has achieved a good level of performance during the study period, driven by technical efficiency. Since 2014, pure technical efficiency (PE) has been >1, indicating that the tourism industry in China has entered a stage of change and promotion; (3) significant spatial differences exist in tourism GTFP in China. For example, the overall pattern of being strongest in the east and weakest in the west has not changed. Currently, eastern, central, and western regions in China rely on different dynamic mechanisms to promote tourism green development. In addition, some provinces have become the core or secondary growth poles of tourism green development in China.

Assessment of aneuploidy formation in human blastocysts resulting from cryopreserved donor eggs.

BACKGROUND: Increased embryo implantation rates were reported after transfer of euploid embryos selected by preimplantation genetic screening (PGS). Egg cryopreservation by vitrification has become one of the most important assisted human reproduction technologies. Although reports indicate that development and implantation of human embryos derived from frozen donor eggs are comparative to fresh eggs, it is still unknown whether egg vitrification increases chromosomal abnormalities in eggs, which in turn causes formation of embryonic aneuploidy. Therefore, in this study, we evaluated the aneuploidy formation in the blastocysts derived from frozen donor eggs and also evaluated the efficiency of egg vitrification as an advanced technology for egg cryopreservation. RESULTS: In this study, donated human eggs from young women were cryopreserved by vitrification and PGS was performed in the resulted blastocysts by DNA microarray. A total of 764 frozen eggs from 75 egg thawing cycles were warmed and 38 blastocysts were biopsied for PGS before embryo transfer. A 97.1% of egg survival rate was obtained and 59.1% of embryos developed to blastocyst stage. After biopsy and PGS, it was found that 84.2% of blastocysts were euploid and 15.8% were aneuploid. Aneuploidy rates varied among donors. Transfers of blastocysts without PGS resulted in higher clinical pregnancy and implantation rates as compared with transfer of blastocysts with PGS. CONCLUSIONS: Although the overall aneuploidy rate was low in the blastocysts derived from frozen donor eggs, high aneuploidy rates were observed in the embryos resulting from some donated eggs. Clinical pregnancy rate was not improved by PGS of embryos resulting from donor eggs, indicating that PGS may not be necessary for embryos derived from donor eggs in most cases.

Multipoint subcutaneous injection of long-acting glucocorticid as a cure for pretibial myxedema.

BACKGROUND: Pretibial myxedema (PM) is an uncommon and characteristic manifestation of Graves' disease (GD), with local autoimmune reaction in cutaneous tissue. The treatment of PM is a clinical challenge. We herein report a patient with PM who achieved complete remission by multipoint subcutaneous injection of long-acting glucocorticid. SUMMARY: A 38-year-old man presented with an 18-month history of GD and a 1-year history of PM. Physical examination revealed mildly prominent eyes, diffuse enlargement of the thyroid gland, and PM of both lower extremities. The patient was treated with triamcinolone acetonide by multipoint subcutaneous injections in a combined dose of 20 mg in each lower extremity administered every 25-28 days. The injection was started from the borderline of the lesions and normal skin by selecting 4 to 5 points per leg for each course and then moving to other parts in subsequent courses of treatment. The depth of needle insertion was 0.5-1.0 cm. The patient's PM achieved complete remission in both lower extremities after an approximately 6-month period that included seven courses of treatment with a total dosage of 280 mg triamcinolone acetonide. CONCLUSIONS: Our experience with this patient suggests that multipoint subcutaneous injection of long-acting glucocorticid is a safe, effective, and convenient treatment of PM in patients with GD.