Codelivery of Doxorubicin and p53 Gene by ß-Cyclodextrin-Based Supramolecular Nanoparticles Formed via Host-Guest Complexation and Electrostatic Interaction.

We developed a supramolecular system for codelivery of doxorubicin (Dox) and p53 gene based on a ß-CD-containing star-shaped cationic polymer. First, a star-shaped cationic polymer consisting of a ß-CD core and 3 arms of oligoethylenimine (OEI), named CD-OEI, was used to form a supramolecular inclusion complex with hydrophobic Dox. The CD-OEI/Dox complex was subsequently used to condense plasmid DNA via electrostatic interactions to form CD-OEI/Dox/DNA polyplex nanoparticles with positive surface charges that enhanced the cellular uptake of both Dox and DNA. This supramolecular drug and gene codelivery system showed high gene transfection efficiency and effective protein expression in cancer cells. The codelivery of Dox and DNA encoding the p53 gene resulted in reduced cell viability and enhanced antitumor effects at low Dox concentrations. With its enhanced cellular uptake and anticancer efficacy, the system holds promise as a delivery carrier for potential combination cancer therapies.

Impacts of biochar aging on its interactions with As(III) and the combined cytotoxicity.

Despite the extensive use of biochar (BC) in soil and aqueous media for pollutant immobilization, the environmental behaviors and health risks of aged BC with multiple pollutants, especially with metal ions possessing various valence states, remain unexplored. Here, we prepared fresh banana peel BC (BP-BC) and aged BP-BCs by acidification (ABP-BC) and oxidation (OBP-BC). ABP-BC was then chosen to explore its environmental behaviors (i.e., adsorption, desorption, and arsenic valence transfer) towards As(III)-Cu(II) and the combined cytotoxicity of BCs with As(III)-Cu(II) was investigated in Human Gastric epithelium cells (GES-1). Our results demonstrate that the aging process notably alters the physicochemical properties of BP-BC, including surface morphology, elemental composition, and surface functional groups, which are key factors affecting the long-term environmental behaviors of BC with As(III)/Cu(II). Specifically, the aging process significantly enhanced the adsorption of As(III) on BC but reduced the adsorption of Cu(II). Although the oxidation of As(III) to As(V) did not change much, the aging process improved the stability of ABP-BC-metal ion complexes, alleviating the release of As(III) in acidic solution. Consequently, the combined cytotoxicity induced by ABP-BC-As(III)-Cu(II) was reduced compared to BP-BC-As(III)-Cu(II). The study highlights the critical roles of the aging process in regulating the As(III) adsorption/desorption dynamics on BCs and their combined cytotoxicity in the presence of multiple metal ions.

Sequencing of the ZMYND15 gene in a cohort of infertile Chinese men reveals novel mutations in patients with teratozoospermia.

BACKGROUND: The information of ZMYND15 in human reproduction is very limited, resulting in the unclear link between ZMYND15 variants and male infertility. METHODS: Whole exome sequencing and Sanger sequencing to identify the potential pathogenic variation of ZMYND15 in infertile men, Papanicolaou staining and electron microscopy to investigate the spermatozoa morphology, western blotting and immunofluorescence staining to confirm the pathogenicity of the identified variants, and proteomic analysis and coimmunoprecipitation to clarify the potential molecular mechanism. RESULTS: A total of 31 ZMYND15 variants were identified in 227 infertile patients. Three deleterious biallelic variants, including a novel compound heterozygous variant of c.1105delG (p.A369Qfs*15) and c.1853T>C (p.F618S), a new homozygous splicing mutation of c.1297+5G>A and a reported homozygous nonsense mutation of c.1209T>A (p.Y403*), were detected in three affected individuals with oligoasthenoteratozoospermia, showing a biallelic pathogenic mutation frequency of 1.3% (3/227). No biallelic pathogenic mutation was found in 692 fertile men. Morphology analysis showed abnormalities in sperm morphology in the patients harbouring ZMYND15 mutations. Western blotting and immunofluorescence staining confirmed the nearly absent ZMYND15 expression in the sperm of the patients. Mechanistically, ZMYND15 might regulate spermatogenesis by interacting with key molecules involved in sperm development, such as DPY19L2, AKAP4 and FSIP2, and might also mediate the expression of the autophagy-associated protein SPATA33 to maintain sperm individualisation and unnecessary cytoplasm removal. CONCLUSION: Our findings broaden the variant and phenotype spectrum of ZMYND15 in male infertility, and reveal the potential signalling pathway of ZMYND15 regulating spermatogenesis, finally confirming the essential role of ZMYND15 in human fertility.

FSIP2 plays a role in the acrosome development during spermiogenesis.

BACKGROUND: Loss-of-function mutations in FSIP2 result in multiple morphological abnormalities of the flagella in humans and mice. Intriguingly, a recent study found that FSIP2 might regulate the expression of acrosomal proteins, indicating that Fsip2 might be involved in acrosome development in mice. However, whether FSIP2 also function in acrosome biogenesis in humans is largely unknown, and the underlying mechanism of which is unexplored. OBJECTIVE: Our objective was to reveal potential function of FSIP2 in regulating sperm acrosome formation. METHODS: We performed whole exome sequencing on four asthenoteratozoospermic patients. Western blot analysis and immunofluorescence staining were conducted to assess the protein expression of FSIP2. Proteomics approach, liquid chromatography-tandem mass spectrometry and co-immunoprecipitation were implemented to clarify the molecules in acrosome biogenesis regulated by FSIP2. RESULTS: Biallelic FSIP2 variants were identified in four asthenoteratozoospermic individuals. The protein expression of MUT-FSIP2 was sharply decreased or absent in vitro or in vivo. Interestingly, aside from the sperm flagellar defects, the acrosomal hypoplasia was detected in numerous sperm from the four patients. FSIP2 co-localised with peanut agglutinin in the acrosome during spermatogenesis. Moreover, FSIP2 interacted with proteins (DPY19L2, SPACA1, HSP90B1, KIAA1210, HSPA2 and CLTC) involved in acrosome biogenesis. In addition, spermatozoa from patients carrying FSIP2 mutations showed downregulated expression of DPY19L2, ZPBP, SPACA1, CCDC62, CCIN, SPINK2 and CSNK2A2. CONCLUSION: Our findings unveil that FSIP2 might involve in sperm acrosome development, and consequently, its mutations might contribute to globozoospermia or acrosomal aplasia. We meanwhile first uncover the potential molecular mechanism of FSIP2 regulating acrosome biogenesis.

Soluble E-cadherin participates in BLM-induced pulmonary fibrosis by promoting EMT and lung fibroblast migration.

Soluble E-cadherin (sE-cad) is an 80 kDa fragment derived from E-cadherin that is shed from the cell surface through proteolytic cleavage and is a biomarker in various cancers that promotes invasion and migration. Alveolar epithelial destruction, aberrant lung fibroblast migration and inflammation contribute to pulmonary fibrosis. Here, we hypothesized that E-cadherin plays an important role in lung fibrosis. In this study, we found that E-cadherin was markedly increased in the bronchoalveolar lavage fluid (BALF) and serum of mice with pulmonary fibrosis and that blocking sE-cad with HECD-1, a neutralizing antibody targeting the ectodomain of E-cadherin, effectively inhibited myofibroblast accumulation and collagen deposition in the lungs after bleomycin (BLM) exposure. Moreover, transforming growth factor-ß (TGF-ß1) induced the shedding of sE-cad from A549 cells, and treatment with HECD-1 inhibited epithelial-mesenchymal transition (EMT) stimulated by TGF-ß1. Fc-E-cadherin (Fc-Ecad), which is an exogenous form of sE-cad, robustly promoted lung fibroblast migration. E-cadherin participates in bleomycin (BLM)-induced lung fibrosis by promoting EMT in the alveolar epithelium and fibroblast activation. E-cadherin may be a novel therapeutic target for lung fibrosis.

Design, synthesis, and analgesia evaluation of novel Transient Receptor Potential Vanilloid 1 (TRPV1) agonists modified from Cannabidiol (CBD).

Pain-relief is a long-term research hotspot with huge demand in clinical treatment. The analgesics currently used have several side effects, such as being addictive and causing gastrointestinal bleeding. Therefore, new drugs and targets in analgesic field are both desirable. Transient Receptor Potential Vanilloid 1 (TRPV1) plays an essential role in pain perception and regulation, providing a new strategy for the development of antinociceptive agents. Here, a series of novel TRPV1 agonists were designed and synthesized based on Cannabidiol (CBD), a widely used pain-relieving agent with weak agonistic activity on TRPV1. According to the results of systematic in vitro and in vivo biological assays, compound 10f was finally identified as a promising TRPV1 agonist, with higher target affinity, stronger analgesic activity, and weak side effect of hyperthermia. Molecular docking simulations revealed a significant hydrogen bond interaction between 10f and Arg557, an amino acid residue key to the activity of TRPV1 protein. Taken together, compound 10f can be used as a lead compound for further optimization.

First report of Watermelon silver mottle orthotospovirus infecting Siraitia grosvenorii in China.

Siraitia grosvenorii, known as "Luohanguo or monk fruit", is a perennial vine belonging to the family Cucurbitaceae. It is cultivated for its fruits, which are used as a Chinese traditional medicine to treat throat, lung and intestine ailments, or as raw material to extract sweet cucurbitane-glycosides as sugar substitute sweeteners (Chen et al., 2007). The production of S. grosvenorii is limited by viral diseases especially cucumber green mottle mosaic virus (CGMMV), papaya ringspot virus (PRSV), watermelon mosaic virus, and zucchini yellow mosaic virus (Liao et al., 2005; Xie et al., 2020). In 2022, virus-like disease consisting of leaf mottling, crinkling, and ringspot was observed on S. grosvenorii plants grown in an insect-proof greenhouse in Guilin City, Guangxi Province, China, with an incidence rate of ~17%. High-throughput sequencing (HTS) was applied to identify potential viruses in the diseased plants. Briefly, total RNA was extracted from a pool of 28 leaf samples (with or without symptoms) of S. grosvenorii using Trizol reagent according to manufacturer's instructions (Invitrogen, U.S.A.). The rRNA was depleted (Epicentre Ribo-zero™ rRNA Removal Kit, Epicentre, U.S.A.), before steps of cDNA library construction (NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®, NEB, U.S.A.), and sequencing (Hiseq 4000 platform, Illumina, U.S.A.). The subsequent bioinformatics analyses were performed according to Liu et al. (2021). HTS of the sample and raw reads processing resulted in 8.4 Gb clean data. The clean reads (150 bp) were de novo assembled into 87,414 contigs (≥200 bp), using CLC Genomics Workbench 21 (Qiagen, Germany). The contigs were annotated by local BLASTX, resulting in matches to CGMMV, PRSV, and watermelon silver mottle virus (WSMoV). Three contigs of 6,557 bp, 4,950 bp, and 3,594 bp were most identical to L (GenBank accession no. JX177647), M (MW051789), and S (KM242056) segments of WSMoV. The complete genome sequences corresponding to the contigs derived from the sample (designated as GL-1 variant of WSMoV, OQ401466-OQ401468) were obtained by reads mapping to segments of these isolates. The reads coverage was ≥99.75% in each RNA segment and the depth of the coverage was in a range of 74-285. To detect the presence of GL-1 in S. grosvenorii plants, three primer pairs D7280F/D7382R (5'-TGATAGCCTGATGAACACCA/5'-TGTCTCTAAACCTTCTACCGC, Tm = 55℃, product size 172 bp), D4512F/D4703R (5'-GCATTGAACTCGCTCACAC/5'-AGTAGACGACCCTGAAGACCT, Tm = 55℃, 192 bp), and D109F/D451R (5'-TTATGGCACAAGAGACAACAGAG/5'-GGGCGTTATGTTCAGTATATTGG, Tm = 56℃, 342 bp) were designed in the L, M, and S segments, respectively. Fresh symptomatic and asymptomatic leaf tissues (n=38) were collected from three fields and their extracted nucleic acids were individually tested with the primers designed by two-steps RT-PCR using TaKaRa RNA PCR kit Ver.3.0 (Takara, Japan). Expected amplicons were obtained in symptomatic samples (n=7) showing mottling, crinkling, and chlorosis. Other samples (n=31) with or without symptoms were negative to WSMoV infection. The amplicons were sequenced, and the sequences obtained shared >99% nt identities with the corresponding GL-1 sequences in GenBank. This is the first report of WSMoV on S. grosvenorii, which provides the basic information for virus disease management.

Association between career adaptability and turnover intention among nursing assistants: the mediating role of psychological capital.

BACKGROUND: High turnover intention of nursing assistants was detrimental to the sustainability of long-term care. Career adaptability is an important determinant in reducing turnover intention, but little research has explored the mechanism from the perspective of psychological capital. The aim of this study was to analyze the association between career adaptability and turnover intention and to examine the mediating role of psychological capital between career adaptability and turnover intention among nursing assistants in mainland China. METHODS: A cross-sectional online study was conducted among 276 nursing assistants from eight nursing homes in Nanjing, China. The participants' career adaptability, psychological capital, and turnover intention were obtained. SPSS 26.0 and Amos 24.0 software were employed for statistical analysis. RESULTS: Career adaptability was positively related to psychological capital and negatively linked to turnover intention (P < 0.01). Psychological capital played a fully mediating role (ß = -0.085, P < 0.05) in the relationship between career adaptability and turnover intention, and the largest indirect effect was generated through the curiosity dimension. CONCLUSIONS: The management of long-term care facilities should focus on assessing the level of career adaptability of nursing assistants. The overall improvement of career adaptability and psychological capital is conducive in reducing turnover intention. Targeted interventions are recommended to improve career adaptability and reduce turnover intentions by increasing career curiosity. Online career adaptability programs can be developed for nursing assistant students to improve their psychological capital and facilitate career transitions.

TNFα-induced abnormal activation of TNFR/NF-κB/FTH1 in endometrium is involved in the pathogenesis of early spontaneous abortion.

Early spontaneous abortion (ESA) is one of the most common complications during pregnancy and the inflammation condition in uterine environment such as long-term exposure to high TNFα plays an essential role in the aetiology. Ferritin heavy chain (FTH1) is considered to be closely associated with inflammation and very important in normal pregnancy, yet the underlying mechanism of how TNFα induced abortion and its relationship with FTH1 remain elusive. In this study, we found that TNFα and FTH1 were positively expressed in decidual stromal cells and increased significantly in the ESA group compared with the normal pregnancy group (NP group). Besides, TNFα expression was positively correlated with FTH1 expression. Furthermore, in vitro cell model demonstrated that high TNFα could induce the abnormal signals of TNFR/NF-κB/FTH1 and activate apoptosis both in human endometrium stromal cells (hESCs) and in local decidual tissues. Taken together, the present findings suggest that the excessive apoptosis in response to TNFα-induced upregulation of FTH1 may be responsible for the occurrence of ESA, and thus provide a possible therapeutic target for the treatment of ESA.

Rapid and simultaneous detection of multiple pathogens in the lower reproductive tract during pregnancy based on loop-mediated isothermal amplification-microfluidic chip.

BACKGROUND: Female reproductive tract infection (RTI) is the common source of varied diseases, especially as an important risk factor for pregnancy outcomes, therefore the rapid, accurate and simultaneous detection of multiple pathogens is in urgent need for assisting the diagnosis and treatment of RTI in pregnant women. Streptococcus agalactiae (S. agalactiae), Enterococcus faecalis (E. faecalis), Gardnerella vaginalis (G. vaginalis), Candida albicans (C. albicans) and Chlamydia trachomatis (C. trachomatis) are five main pathogens in lower genital tract with high risk, serious consequences and clinical demands. The combination of loop-mediated isothermal amplification (LAMP) and microfluidic technology was used to develop the LAMP-microfluidic chip for rapid, simple, sensitive and simultaneous detection of the five target pathogens above. RESULTS: Standard strains and clinical isolates were used for the establishment of the novel LAMP method in tube and LAMP-microfluidic chip, followed by the chip detection on 103 clinical samples and PCR verification partially. The sensitivities of LAMP of S. agalactiae, E. faecalis, G. vaginalis, and C. albicans in tube were 22.0, 76.0, 13.2, 1.11 CFU/µL, respectively, and C. trachomatis was 41.3 copies/µL; on LAMP-microfluidic chip they were 260, 154, 3.9 and 7.53 CFU/µL, respectively, and C. trachomatis was 120 copies/µL. The positive coincidence rates of clinical stains in tube and on chip experiments were 100%. Compared with the classic culture method performed in hospitals, the positive coincidence rate of the 103 clinical samples detected by LAMP-microfluidic chip were 100%. For the six inconsistent ones, including four G. vaginalis and two C. albicans positive samples tested by LAMP-microfluidic chip and verified by PCR were negative by culturing method in hospitals, indicating the lack of efficient detection by the classic culturing method. CONCLUSION: Our study suggested that the LAMP-microfluidic chips could simultaneously, efficiently, and accurately detect multiple main pathogens, including S. agalactiae, E. faecalis, G. vaginalis, C. albicans and C. trachomatis, in clinical samples of female RTI to give a great clinical value. Accordingly, this novel method has the potential to provide a valuable reference for female RTI screening and early diagnosis during pregnancy.

A loss-of-function variant in SSFA2 causes male infertility with globozoospermia and failed oocyte activation.

Globozoospermia (OMIM: 102530) is a rare type of teratozoospermia (< 0.1%). The etiology of globozoospermia is complicated and has not been fully revealed. Here, we report an infertile patient with globozoospermia. Variational analysis revealed a homozygous missense variant in the SSFA2 gene (NM_001130445.3: c.3671G > A; p.R1224Q) in the patient. This variant significantly reduced the protein expression of SSFA2. Immunofluorescence staining showed positive SSFA2 expression in the acrosome of human sperm. Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and Coimmunoprecipitation (Co-IP) analyses identified that GSTM3 and Actin interact with SSFA2. Further investigation revealed that for the patient, regular intracytoplasmic sperm injection (ICSI) treatment had a poor prognosis. However, Artificial oocyte activation (AOA) by a calcium ionophore (A23187) after ICSI successfully rescued the oocyte activation failure for the patient with the SSFA2 variant, and the couple achieved a live birth. This study revealed that SSFA2 plays an important role in acrosome formation, and the homozygous c.3671G > A loss-of-function variant in SSFA2 caused globozoospermia. SSFA2 may represent a new gene in the genetic diagnosis of globozoospermia, especially the successful outcome of AOA-ICSI treatment for couples, which has potential value for clinicians in their treatment regimen selections.

Synthesis and Characterization of Palmitoyl-block-poly(methacryloyloxyethyl phosphorylcholine) Polymer Micelles for Anticancer Drug Delivery.

We report the synthesis and characterization of an amphiphilic polymer comprising a hydrophobic palmitoyl (Pal) group and a zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (pMPC) block, which is capable of forming micelles as a drug carrier system for delivering hydrophobic anticancer drugs such as doxorubicin (DOX). We hypothesize that the sharp polarity contrast between the Pal domain and the pMPC block would strengthen the micelles and improve the drug loading capacity, while the pMPC shells improve the micelle stability and cellular uptake efficiency. In this study, the Pal-pMPC polymer was characterized and compared with a Pal-poly(ethylene glycol) (Pal-PEG) polymer in terms of their micelle formation, cytotoxicity, and drug loading of DOX. The DOX-loaded Pal-pMPC micelles were further evaluated for the cellular uptake and anticancer activities in cell culture systems including the non-multidrug-resistance HeLa cell line and the multidrug-resistance AT3B-1 cell line. The results showed that the Pal-pMPC polymer had a minimal toxicity. The Pal-pMPC micelles exhibited higher drug loading capacity and enhanced cellular internalization efficiency compared to micelles formed by the Pal-PEG polymer. It was also found that DOX-loaded Pal-pMPC micelles exhibited a more efficient anticancer effect than Pal-PEG micelles in multidrug-resistance cancer cells in an environment with fetal bovine serum.

High-strength deep learning image reconstruction in coronary CT angiography at 70-kVp tube voltage significantly improves image quality and reduces both radiation and contrast doses.

OBJECTIVES: To explore the use of 70-kVp tube voltage combined with high-strength deep learning image reconstruction (DLIR-H) in reducing radiation and contrast doses in coronary CT angiography (CCTA) in patients with body mass index (BMI) < 26 kg/m2, in comparison with the conventional scan protocol using 120 kVp and adaptive statistical iterative reconstruction (ASIR-V). METHODS: A total of 100 patients referred to CCTA were prospectively enrolled and randomly divided into two groups: low-dose group (n = 50) with 70 kVp, Smart mA for noise index (NI) of 36HU, contrast dose rate of 16mgI/kg/s, and DLIR-H, and conventional group (n = 50) with 120 kV, Smart mA for NI of 25HU, contrast dose rate of 32mgI/kg/s, and 60%ASIR-V. Radiation and contrast dose, subjective image quality score, and objective image quality measurement (image noise, contrast-noise-ratio (CNR), and signal-noise-ratio (SNR) for vessel) were compared between the two groups. RESULTS: Low-dose group used significantly reduced contrast dose (23.82 ± 3.69 mL, 50.6% reduction) and radiation dose (0.75 ± 0.14 mSv, 54.5% reduction) compared to the conventional group (48.23 ± 6.38 mL and 1.65 ± 0.66 mSv, respectively) (all p < 0.001). Both groups had similar enhancement in vessels. However, the low-dose group had lower background noise (23.57 ± 4.74 HU vs. 35.04 ± 8.41 HU), higher CNR in RCA (48.63 ± 10.76 vs. 29.32 ± 5.52), LAD (47.33 ± 10.20 vs. 29.27 ± 5.12), and LCX (46.74 ± 9.76 vs. 28.58 ± 5.12) (all p < 0.001) compared to the conventional group. CONCLUSIONS: The use of 70-kVp tube voltage combined with DLIR-H for CCTA in normal size patients significantly reduces radiation dose and contrast dose while further improving image quality compared with the conventional 120-kVp tube voltage with 60%ASIR-V. KEY POINTS: • The combination of 70-kVp tube voltage and high-strength deep learning image reconstruction (DLIR-H) algorithm protocol reduces approximately 50% of radiation and contrast doses in coronary computed tomography angiography (CCTA) compared with the conventional scan protocol. • CCTA of normal size (BMI < 26 kg/m2) patients acquired at sub-mSv radiation dose and 24 mL contrast dose through the combination of 70-kVp tube voltage and DLIR-H algorithm achieves excellent diagnostic image quality with a good inter-rater agreement. • DLIR-H algorithm shows a higher capacity of significantly reducing image noise than adaptive statistical iterative reconstruction algorithm in CCTA examination.

Whole-exome sequencing of a cohort of infertile men reveals novel causative genes in teratozoospermia that are chiefly related to sperm head defects.

STUDY QUESTION: Can whole-exome sequencing (WES) and in vitro validation studies identify new causative genes associated with teratozoospermia, particularly for sperm head defect? SUMMARY ANSWER: We investigated a core group of infertile patients, including 82 cases with unexplained abnormal sperm head and 67 individuals with multiple morphological abnormalities of the sperm flagella (MMAF), and revealed rare and novel deleterious gene variants correlated with morphological abnormalities of the sperm head or tail defects. WHAT IS KNOWN ALREADY: Teratozoospermia is one of the most common factors causing male infertility. Owing to high phenotypic variability, currently known genetic causes of teratozoospermia can only explain a rather minor component for patients with anomalous sperm-head shapes, and the agents responsible for atypical sperm head shapes remain largely unknown. STUDY DESIGN, SIZE, DURATION: We executed WES analysis of a Chinese cohort of patients (N = 149) with teratozoospermia to identify novel genetic causes particularly for defective sperm head. We also sought to reveal the influence of different abnormalities of sperm morphology on ICSI outcome. PARTICIPANTS/MATERIALS, SETTING, METHODS: In this study, a cohort of 149 infertile men (82 with abnormal sperm head and 67 with MMAF) were recruited. We implemented WES on infertile patients and analyzed the negative effects of the mutations of candidate genes on their protein conformations and/or expression. We also investigated the candidate genes' spatiotemporal expression/localization during spermatogenesis in both humans and mice, and explored their interactions with proteins that are known to be involved in sperm development. We also compared the ICSI outcomes of the affected individuals with various aberrations in sperm morphology. MAIN RESULTS AND THE ROLE OF CHANCE: We identified rare and deleterious variants of piwi like RNA-mediated gene silencing 4 (PIWIL4: 1/82 patients, 1.21%), coiled-coil and C2 domain containing 1B (CC2D1B: 1/82 patients, 1.21%), cyclin B3 (CCNB3: 1/82 patients, 1.21%), KIAA1210 (KIAA1210: 2/82 patients, 2.43%) and choline phosphotransferase 1 (CHPT1: 1/82 patients, 1.21%), which are novel correlates of morphological abnormalities of the sperm head; functional evidence supports roles for all of these genes in sperm head formation. The mutations of septin 12 (SEPTIN12: 2/82 patients, 2.43%) are suggested to be associated with acrosome defects. We additionally observed novel causative mutations of dynein axonemal heavy chain 2 (DNAH2: 1/67 patients, 1.49%), dynein axonemal heavy chain 10 (DNAH10: 1/67 patients, 1.49%) and dynein axonemal heavy chain 12 (DNAH12: 1/67 patients, 1.49%) in patients with MMAF, and revealed a significantly lower fertilization rate of the abnormal sperm-head group compared to the MMAF group following ICSI. Consequently, our study also suggests that the mutations of PIWIL4 and CC2D1B might be circumvented by ICSI to a degree, and that CHPT1 and KIAA1210 loss-of-function variants might be associated with failed ICSI treatment. LIMITATIONS, REASONS FOR CAUTION: In this study, we discovered the relationship between the genotype and phenotype of the novel causative genes of sperm head deformities in humans. However, the molecular mechanism of the relevant genes involved in sperm head development needs to be further illuminated in future research. Furthermore, evidence should be provided using knockout/knock-in mouse models for additional confirmation of the roles of these novel genes in spermatogenesis. WIDER IMPLICATIONS OF THE FINDINGS: This cohort study of 149 Chinese infertile men documents novel genetic factors involved in teratozoospermia, particularly in anomalous sperm head formation. For the first time, we suggest that SEPTIN12 is related to human acrosomal hypoplasia, and that CCNB3 is a novel causative gene for globozoospermia in humans. We also uncovered variants in two genes-KIAA1210 and CHPT1associated with acrosomal biogenesis in patients with small or absent acrosomes. Additionally, it is postulated that loss-of-function mutations of PIWIL4 and CC2D1B have a contribution to the abnormal sperm-head formation. Furthermore, we are first to demonstrate the influence of different sperm morphologies on ICSI outcomes and indicates that the abnormal sperm head may play a significant role in fertilization failure. Our findings therefore provide valuable information for the diagnosis of teratozoospermia, particularly with respect to abnormalities of the sperm head. This will allow clinicians to adopt the optimal treatment strategy and to develop personalized medicine directly targeting these effects. STUDY FUNDING/COMPETING INTEREST(S): This work was financed by the West China Second University Hospital of Sichuan University (KS369 and KL042). The authors declare that they do not have any conflicts of interests. TRIAL REGISTRATION NUMBER: N/A.

ß-Cyclodextrin-Polyacrylamide Hydrogel for Removal of Organic Micropollutants from Water.

Water pollution by various toxic substances remains a serious environmental problem, especially the occurrence of organic micropollutants including endocrine disruptors, pharmaceutical pollutants and naphthol pollutants. Adsorption process has been an effective method for pollutant removal in wastewater treatment. However, the thermal regeneration process for the most widely used activated carbon is costly and energy-consuming. Therefore, there has been an increasing need to develop alternative low-cost and effective adsorption materials for pollutant removal. Herein, ß-cyclodextrin (ß-CD), a cheap and versatile material, was modified with methacrylate groups by reacting with methacryloyl chloride, giving an average degree of substitution of 3 per ß-CD molecule. ß-CD-methacrylate, which could function as a crosslinker, was then copolymerized with acrylamide monomer via free-radical copolymerization to form ß-CD-polyacrylamide (ß-CD-PAAm) hydrogel. Interestingly, in the structure of the ß-CD-PAAm hydrogel, ß-CD is not only a functional unit binding pollutant molecules through inclusion complexation, but also a structural unit crosslinking PAAm leading to the formation of the hydrogel 3D networks. Morphological studies showed that ß-CD-PAAm gel had larger pore size than the control PAAm gel, which was synthesized using conventional crosslinker instead of ß-CD-methacrylate. This was consistent with the higher swelling ratio of ß-CD-PAAm gel than that of PAAm gel (29.4 vs. 12.7). In the kinetic adsorption studies, phenolphthalein, a model dye, and bisphenol A, propranolol hydrochloride, and 2-naphthol were used as model pollutants from different classes. The adsorption data for ß-CD-PAAm gel fitted well into the pseudo-second-order model. In addition, the thermodynamic studies revealed that ß-CD-PAAm gel was able to effectively adsorb the different dye and pollutants at various concentrations, while the control PAAm gel had very low adsorption, confirming that the pollutant removal was due to the inclusion complexation between ß-CD units and pollutant molecules. The adsorption isotherms of the different dye and pollutants by the ß-CD-PAAm gel fitted well into the Langmuir model. Furthermore, the ß-CD-PAAm gel could be easily recycled by soaking in methanol and reused without compromising its performance for five consecutive adsorption/desorption cycles. Therefore, the ß-CD-PAAm gel, which combines the advantage of an easy-to-handle hydrogel platform and the effectiveness of adsorption by ß-CD units, could be a promising pollutant removal system for wastewater treatment applications.

Surface Charge Switchable Polymer/DNA Nanoparticles Responsive to Tumor Extracellular pH for Tumor-Triggered Enhanced Gene Delivery.

A tumor-targeted surface charge switchable polymeric gene delivery system with the function of switching surface charge upon reaching the tumor site owing to the tumor extracellular pH (pHe) was developed. The delivery system was fabricated by two steps. First, the positively charged polyplexe nanoparticles were formed between ß-cyclodextrin-oligoethylenimine star polymer (CD-OEI) and plasmid DNA (pDNA). Next, the CD-OEI/pDNA polyplex nanoparticles were coated with a pHe-responsive anionic polymer via an electrostatic interaction to form ternary complexes. The pHe-responsive anionic polymer was block copolymers of poly(ethylene glycol) (PEG) and poly(2-aminoethyl methacrylate) (pAEMA) modified with 2,3-dimethylmaleic anhydride (denoted as PPD). The coating polymer was mixed with a small amount of pHe-insensitive PEG-pAEMA modified with succinic anhydride (denoted as PPS), giving a balanced negatively charged and PEG-shielded surface with a pHe-responsive property for achieving the expected tumor-triggered enhanced gene delivery. At physiological pH 7.4, owing to the charge shielding of anionic surface coating and the PEGylation, the negatively charged CD-OEI/pDNA/PPD+PPS polyplex complexes could avoid the undesirable interaction with serum proteins and nontargeted components. However, the amide bond of PPD was sensitive to pH changes and could be easily hydrolyzed under acidic pHe (<6.8) to expose the primary amine group due to nucleophile catalysis by the carboxylic acid. The PEG block in the copolymers was used to further enhance the surface-shielding effect. Our data showed that excellent particle salt stability and serum tolerance were achieved through the PPD+PPS surface coating. The CD-OEI/pDNA/PPD+PPS complexes achieved lower cellular uptake and transfection efficiency at neutral pH 7.4 while exhibiting comparable cellular uptake and transfection efficiency at acidic pH 6.5 as compared to the uncoated polyplexes, indicating that the surface charge switching worked well.

Thermoresponsive Hydrogel Induced by Dual Supramolecular Assemblies and Its Controlled Release Property for Enhanced Anticancer Drug Delivery.

Supramolecular hydrogels based on inclusion complexation between cyclodextrins (CDs) and polymers have attracted much interest because of their potential for biomedical applications. It is also attractive to incorporate stimuli-responsive properties into the system to create "smart" hydrogels. Herein, a poly(N-isopropylacrylamide) (PNIPAAm) star polymer with a ß-CD core and an adamantyl-terminated poly(ethylene glycol) (Ad-PEG) polymer were synthesized. They self-assembled into a thermoresponsive pseudo-block copolymer through host-guest complexation and formed supramolecular micelles with the change in environment temperature. Subsequently, an injectable polypseudorotaxane-based supramolecular hydrogel was formed between α-CD and the PEG chains of the pseudo-block copolymer. The hydrogel had a unique network structure involving two types of supramolecular self-assemblies between cyclodextrins and polymers, that is, the host-guest complexation between ß-CD units and adamantyl groups and the polypseudorotaxane formation between α-CD and PEG chains. We hypothesize that the dual supramolecular hydrogel formed at room temperature may be enhanced by increasing the temperature over the lower critical solution temperature of PNIPAAm because of the hydrophobic interactions of PNIPAAm segments. Furthermore, if the hydrogel is applied for sustained delivery of hydrophobic drugs, the copolymer dissolved from the hydrogel could micellize and continue to serve as micellar drug carriers with the drug encapsulated in the hydrophobic core. Rheological tests revealed that the hydrophobic interactions of the PNIPAAm segments could significantly enhance the strength of the hydrogel when the temperature increased from 25 to 37 °C. As compared to hydrogels formed by α-CD and PEG alone, the sustained release property of this thermoresponsive hydrogel for an anticancer drug, doxorubicin (DOX), improved at 37 °C. The hydrogel dissolved slowly and released the pseudo-block copolymer in the form of micelles that continued to serve as drug carriers with DOX encapsulated in the hydrophobic core, achieving a better cellular uptake and anticancer effect than free DOX controls, even in multidrug-resistant cancer cells. According to these findings, the dual supramolecular hydrogel developed in this work with remarkable thermoresponsive properties might have potential for sustained anticancer drug delivery with enhanced therapeutic effect in multidrug-resistant cancer cells.

The SIRT2 inhibitor AK-7 decreases cochlear cell apoptosis and attenuates noise-induced hearing loss.

Oxidative damage plays a critical role in cochlear cell apoptosis, which is central to the physiopathology of noise-induced hearing loss (NIHL). Sirtuin 2 (SIRT2) is an NAD-dependent deacetylase that regulates cellular response to oxidative stress, however, its role in NIHL remains poorly understood. Here, we report that SIRT2 is upregulated in the cochlea after noise exposure. Functionally, the treatment of AK-7, one specific SIRT2 inhibitor, attenuates the progression of NIHL. In addition, AK-7 treatment reduces oxidative nuclear DNA damage and apoptosis in the cochlea after noise exposure. Moreover, AK-7 treatment reduces apoptosis of mouse inner ear HEI-OC1 cells exposed to oxidative stress in vitro. Taken together, these results suggest that SIRT2 inhibition with AK-7 reduces cochlear cell apoptosis through attenuating oxidative stress-induced damage, which may underlie its protective role against NIHL. This study also implies that AK-7 may have potential therapeutic significance in the intervention of NIHL.

Development and Validation of a Nomogram for Predicting Survival in Patients with Thyroid Cancer.

BACKGROUND The AJCC staging system is inadequate for use in patients with thyroid carcinomas. Here, we aimed to establish a nomogram for thyroid cancer, and we compare its prognostic value with the AJCC staging system in adults diagnosed with thyroid carcinoma. MATERIAL AND METHODS Patient records were obtained from the Surveillance, Epidemiology, and End Result database. The 8491 included patients were divided into a modeling cohort (n=5943) and a validation cohort (n=2548). The variables included in the modeling cohort were selected using a backward stepwise selection method with Cox regression, and the prognosis nomogram was constructed. In the validation cohort, we compared our survival model with the AJCC prognosis model using the concordance index, the area under the time-dependent receiver operating characteristic curve, the net reclassification improvement, the integrated discrimination improvement, calibration plotting, and decision curve analysis. RESULTS Twelve independent prognostic factors were identified and used to establish the nomogram. In particular, marital status was included in a survival prediction model of thyroid cancer for the first time. The concordance index, area under the time-dependent receiver operating characteristic curve, net reclassification improvement, integrated discrimination improvement, calibration plotting, and decision curve analysis for the nomogram showed better performance compared to the AJCC staging system. CONCLUSIONS We have developed and validated a highly accurate thyroid cancer prognosis nomogram. The prognostic value of the nomogram is better than that of the AJCC staging system alone.

Prediction of protein-protein interactions by label propagation with protein evolutionary and chemical information derived from heterogeneous network.

Prediction of protein-protein interactions (PPIs) is of great significance. To achieve this, we propose a novel computational method for PPIs prediction based on a similarity network fusion (SNF) model for integrating the physical and chemical properties of proteins. Specifically, the physical and chemical properties of protein are the protein amino acid mutation rate and its hydrophobicity, respectively. The amino acid mutation rate is extracted using a BLOSUM62 matrix, which puts the protein sequence into block substitution matrix. The SNF model is exploited to fuse protein physical and chemical features of multiple data by iteratively updating each original network. Finally, the complementary features from the fused network are fed into a label propagation algorithm (LPA) for PPIs prediction. The experimental results show that the proposed method achieves promising performance and outperforms the traditional methods for the public dataset of H. pylori, Human, and Yeast. In addition, our proposed method achieves average accuracy of 76.65%, 81.98%, 84.56%, 84.01% and 84.38% on E. coli, C. elegans, H. sapien, H. pylori and M. musculus datasets, respectively. Comparison results demonstrate that the proposed method is very promising and provides a cost-effective alternative for predicting PPIs. The source code and all datasets are available at http://pan.baidu.com/s/1dF7rp7N.