Proactive functional classification of all possible missense single-nucleotide variants in KCNQ4.

Clinical exome sequencing has yielded extensive disease-related missense single-nucleotide variants (SNVs) of uncertain significance, leading to diagnostic uncertainty. KCNQ4 is one of the most commonly responsible genes for autosomal dominant nonsyndromic hearing loss. According to the gnomAD cohort, approximately one in 100 people harbors missense variants in KCNQ4 (missense variants with minor allele frequency > 0.1% were excluded), but most are of unknown consequence. To prospectively characterize the function of all 4085 possible missense SNVs of human KCNQ4, we recorded the whole-cell currents using the patch-clamp technique and categorized 1068 missense SNVs as loss of function, as well as 728 loss-of-function SNVs located in the transmembrane domains. Further, to mimic the heterozygous condition in Deafness nonsyndromic autosomal dominant 2 (DFNA2) patients caused by KCNQ4 variants, we coexpressed loss-of-function variants with wild-type KCNQ4 and found 516 variants showed impaired or only partially rescued heterogeneous channel function. Overall, our functional classification is highly concordant with the auditory phenotypes in Kcnq4 mutant mice and the assessments of pathogenicity in clinical variant interpretations. Taken together, our results provide strong functional evidence to support the pathogenicity classification of newly discovered KCNQ4 missense variants in clinical genetic testing.

Rapid quality assessment of Gentianae Macrophyllae Radix based on near-infrared spectroscopy and capillary electrophoresis.

The aim of this study was to establish a rapid quality assessment method for Gentianae Macrophyllae Radix (RGM) using near-infrared (NIR) spectra combined with chemometric analysis. The NIR spectra were acquired using an integrating sphere diffuse reflectance module, using air as the reference. Capillary electrophoresis (CE) analyses were performed on a model P/ACE MDQ Plus system. Partial least squares-discriminant analysis qualitative model was developed to distinguish different species of RGM samples, and the prediction accuracy for all samples was 91%. The CE response values at each retention time were predicted by building a partial least squares regression (PLSR) calibration model with the CE data set as the Y matrix and the NIR spectra data set as the X matrix. The converted CE fingerprints basically match the real ones, and the six main peaks can be accurately predicted. Transforming NIR spectra fingerprints into the form of CE fingerprints increases its interpretability and more intuitively demonstrates the components that cause diversity among samples of different species and origins. Loganic acid, gentiopicroside, and roburic acid were considered quality indicators of RGM and calibration models were built using PLSR algorithm. The developed models gave root mean square error of prediction of 0.2592% for loganic acid, 0.5341% for gentiopicroside, and 0.0846% for roburic acid. The overall results demonstrate that the rapid quality assessment system can be used for quality control of RGM.

Wavelength-Tunable, Long Lifetime, and Biocompatible Luminescent Nanoparticles Based on a Vitamin E-Derived Material for Inflammation and Tumor Imaging.

Luminescence imaging is one of the most effective noninvasive strategies for detection and stratification of inflammation and oxidative stress that are closely related to the pathogenesis of numerous acute and chronic diseases. Herein biocompatible nanoparticles based on a peroxalate ester derived from vitamin E (defined as OVE) are developed. In combination with different fluorophores, OVE can generate luminescence systems with emission wavelengths varying from blue to the near-infrared light in its native and nanoparticle forms, in the presence of hydrogen peroxide (H2 O2 ). The OVE-based nanoprobes exhibit high luminescence signals with extremely long lifetime, upon triggering by inflammatory conditions with abnormally elevated H2 O2 . Activated neutrophils and macrophages can be illuminated by this type of luminescent nanoprobes, with luminescence intensities positively correlated with inflammatory cell counts. In mouse models of peritonitis, alcoholic liver injury, drug-induced acute liver injury, and acute lung injury, the developed luminescence nanoprobes enable precision imaging of inflammation and disease progression. Moreover, tumors expressing a high level of H2 O2 can be shined. Importantly, the OVE-based nanoplatform shows excellent in vitro and in vivo biocompatibility.

Systematical evolution on a Zn-Mg alloy potentially developed for biodegradable cardiovascular stents.

To reduce the long-term side effects of permanent metallic stents, a new generation of cardiovascular stents called "biodegradable stents" is being extensively developed. Zinc has been considered as a promising candidate material for biodegradable cardiovascular stents due to its excellent biocompatibility and appropriate biodegradability. However, weak mechanical properties limit its further clinic application. In this study, hot extruded pure Zn and Zn-0.02 Mg alloy were prepared. Compared with pure Zn, Zn-0.02 Mg alloy showed more homogeneous microstructure, much smaller grain size and higher mechanical strength. Zn-0.02 Mg alloy presented uniform corrosion morphologies during the immersion process, and its corrosion rates was higher than that of pure Zn. Hemocompatibility results showed that the Zn-based alloy had extremely low hemolysis rate (0.74 ± 0.15%) and strong inhibitory effect on blood coagulation, platelet adhesion and aggregation. Zn-0.02 Mg alloy also exhibited excellent cytocompatibility. Its extracts could significantly promote the proliferation of endothelial cells. Moreover, the antibacterial activities of the Zn-based alloy were demonstrated by spread plate assay, live/dead viability assay and bacterial morphology observation. These results indicate that the extruded Zn-0.02 Mg alloy has a potential in biodegradable cardiovascular stents.

Low-Affinity Zinc Sensor Showing Fluorescence Responses with Minimal Artifacts.

The study of the zinc biology requires molecular probes with proper zinc affinity. We developed a low-affinity zinc probe (HBO-ACR) based on an azacrown ether (ACR) and an 2-(2-hydroxyphenyl)benzoxazole (HBO) fluorophore. This probe design imposed positive charge in the vicinity of a zinc coordination center, which enabled fluorescence turn-on responses to high levels of zinc without being affected by the pH and the presence of other transition-metal ions. Steady-state and transient photophysical investigations suggested that such a high tolerance benefits from orchestrated actions of proton-induced nonradiative and zinc-induced radiative control. The zinc bioimaging utility of HBO-ACR has been fully demonstrated with the use of human pancreas epidermoid carcinoma, PANC-1 cells, and rodent hippocampal neurons from cultures and acute brain slices. The results obtained through our studies established the validity of incorporating positively charged ionophores for the creation of low-affinity probes for the visualization of biometals.

Chemical constituents, pharmacological activities and quality evaluation methods of genus Hippocampus: A comprehensive review.

The genus Hippocampus is a multi-origin animal species with high medicinal and healthcare values. About 57 species of Hippocampus spread worldwide, of which about 14 species can be used as medicine, showing anti-oxidation, anti-inflammation, anti-depressant, anti-hypertension, anti-prostatic hyperplasia, antivirus, anti-apoptotic, antifatigue, and so on. And those pharmacological effects are mainly related to their active ingredients, including amino acids, abundant proteins (peptides and oligopeptides), fatty acids, nucleosides, steroids, and other small molecular compounds. The main means of authentication of Hippocampus species are morphological identification, microscopic identification, thin layer chromatography method, fingerprint method and genomics method. This review will provide useful insight for exploration, further study and precise medication of Hippocampus in the future.

DNA binding and anticancer activity of naphthalimides with 4-hydroxyl-alkylamine side chains at different lengths.

A series of novel naphthalimide derivatives modified with various hydroxyl-alkylamines at 4-position have been synthesized. Their DNA binding properties were investigated by UV-Vis, fluoescence, and circular dichroism (CD) spectroscopies and thermal denaturation. The results showed that compounds 3a-e as the DNA intercalator exhibited middle binding affinities with Ct-DNA. The anticancer activities of 3a-e were preliminarily evaluated, compounds 3c and 3e exhibited potent anticancer activities against Bel-7402 cell line with IC(50) values of 5.57 and 9.17µM, respectively. More interestingly, enhancement of the fluorescence emission was found in the complexes of 3a-e with Ct-DNA, especially for 3c. This would make these compounds as potential DNA staining agents.

A Multi-Bioactive Nanomicelle-Based "One Stone for Multiple Birds" Strategy for Precision Therapy of Abdominal Aortic Aneurysms.

Abdominal aortic aneurysm (AAA) remains a lethal aortic disease in the elderly. Currently, no effective drugs can be clinically applied to prevent the development of AAA. Herein, a "one stone for multiple birds" strategy for AAA therapy is reported. As a proof of concept, three bioactive conjugates are designed and synthesized, which can assemble into nanomicelles. Cellularly, these nanomicelles significantly inhibit migration and activation of inflammatory cells as well as protect vascular smooth muscle cells (VSMCs) from induced oxidative stress, calcification and apoptosis, with the best effect for nanomicelles (TPTN) derived from a conjugate defined as TPT. After intravenous delivery, TPTN efficiently accumulates in the aneurysmal tissue of AAA rats, showing notable distribution in neutrophils, macrophages and VSMCs, all relevant to AAA pathogenesis. Whereas three examined nanomicelles effectively delay expansion of AAA in rats, TPTN most potently prevents AAA growth by simultaneously normalizing the pro-inflammatory microenvironment and regulating multiple pathological cells. TPTN is effective even at 0.2 mg kg-1 . Besides, TPTN can function as a bioactive nanoplatform for site-specifically delivering and triggerably releasing anti-aneurysmal drugs, affording synergistic therapeutic effects. Consequently, TPTN is a promising multi-bioactive nanotherapy and bioresponsive targeting delivery nanocarrier for effective therapy of AAA and other inflammatory vascular diseases.

Synthesis and biological activity of novel 5'-arylamino-nucleosides by microwave-assisted one-pot tandem Staudinger/aza-Wittig/reduction.

Novel pseudonucleosides with benzylamino group on 5'-position (4) were synthesized by using the microwave-assisted one-pot tandem Staudinger/aza-Wittig/reduction reaction in good yields of 55.2-71.7%. The deacetylation of 4 afforded compounds 5. HIV-1 reverse transcriptase (RT) inhibitory and antitumor activities were preliminarily evaluated with 5. The results showed that the new pseudonucleosides (5) could effectively inhibit HIV-1 RT activity, but no antitumor activity.

Synthesis and antitumor activity of novel ribonucleosides with C-5 OH replaced by a diaminopyrimidinyl group.

A series of novel ribonucleosides with C-5 OH replaced by a diaminopyrimidinyl group were synthesized by successively nucleophilic substitutions of 5'-deoxy-5'-amino-ribonucleosides with 2,4-dichloropyrimidine and then with various fatty amines under microwave irradiation. Their anticancer activities in vitro were preliminarily evaluated. Compounds 7a and 8a only exhibited anticancer activity against A549 cell line with the IC(50) values of 10.73 and 10.99 µM, respectively. In addition, 7h and 8h showed potent activities against both A549 and Hela cell lines with the IC(50) values of 12.71, 8.55 and 8.44, 5.55 µM, respectively.

Development of a carbamate-conjugated catechol ligand and its application to Cs extraction from contaminated soil by using supercritical CO2.

Extraction of radioactive Cs from contaminated soil is a crucial aspect of remediation after nuclear accidents. For this purpose, we have developed a new type of ligand, carbamate-conjugated catechol, to assist in metal extraction by using supercritical CO2 (SCCO2). The synthesis process for this ligand is relatively simple, and the carbamate-conjugated catechol ligand dissolves well in SCCO2. The measured ligand distribution coefficient increased according to a power law with an exponent of 1.7 as the ligand concentration increased, indicating that approximately two ligand molecules are needed to extract one Cs ion. The roles of additives (ligand, co-ligand, and water) were limited when they were used separately, but the combination of these additives was important. We tested 27 combinations of these three additives for extracting Cs from artificially contaminated sea sand. A quantitative analysis indicated that the ligand had the strongest influence on Cs extraction, followed by water, and the co-ligand. The carbamate-conjugated catechol ligand was then used for Cs extraction from artificially contaminated real soil. Three types of soil were prepared: coarse soil (particle size = 0.5-1 mm), medium soil (particle size = 0.2-0.5 mm), and fine soil (particle size < 0.2 mm). The Cs fractions extracted from the coarse, medium, and fine soils were measured to be 95%, 91% and 70% of the Cs fraction extracted from sea sand, respectively, which indicates the existence of a surface area effect. Additionally, we suspect that Cs undergoes chemical interaction on the surface of real soil.

Corrosion behavior and biocompatibility evaluation of a novel zinc-based alloy stent in rabbit carotid artery model.

Zinc (Zn) and its alloys have been proved to be promising candidate materials for biodegradable cardiovascular stents. In this study, a novel extruded Zn-0.02 Mg-0.02Cu alloy was prepared. Compared with pure Zn, the Zn-based alloy showed higher mechanical properties, and the Zn-based alloy could significantly accelerate Zn2+ release, reaching 0.61 ± 0.11 µg/mL at 15 days of immersion. In vitro biocompatibility studies demonstrated that the Zn-based alloy had excellent cytocompatibility and hemocompatibility, including low hemolysis rate (0.63 ± 0.12%) and strong inhibitory effect on platelet adhesion. Subsequently, the Zn-based alloy stent was implanted into the left carotid arteries of New Zealand white rabbits for 12 months. All the rabbits survived without any adverse clinical events, and all the stented arteries were patent during the study period. Rapid endothelialization at 1 week of implantation was observed, suggesting a low cytotoxicity and thrombosis risk. The stent corroded slowly and no obvious intimal hyperplasia was observed for 6 months, after which corrosion accelerated at 12 months. In addition, no obvious thrombosis and systemic toxicity during implantation period were observed, indicating its potential as the backbone of biodegradable cardiovascular stents. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1814-1823, 2019.

[Pilot study of a cell membrane like biomimetic drug-eluting coronary stent].

A novel bioinspired phospholipid copolymer has been synthesized by the radical polymerization of poly2-Methacryloyloxyethylphosphorylcholine (MPC), stearyl methacrylate (SMA), hydroxypropyl methacrylate (HPMA) and trimethoxysilylpropyl methacrylate (TSMA). Contact angle results indicated that the coating surface rearranged to get a more hydrophilic surface at the polymer/water interface. The membrane mimic phosphorylcholine coating surface could resist the platelet adhesion and prolong plasma recalcification time significantly. Rapamycin was used as model drugs to prepare drug-eluting coating. The animal experiments showed that this novel drug-eluting stent could effectively prevent the phenomena of restenosis.