Intestinal microbiota via NLRP3 inflammasome dependent neuronal pyroptosis mediates anxiety-like behaviour in mice exposed to 3.5 GHz radiofrequency radiation.

The rapid development of 5G communication technology has increased public concern about the potential adverse effects on human health. Till now, the impacts of radiofrequency radiation (RFR) from 5G communication on the central nervous system and gut-brain axis are still unclear. Therefore, we investigated the effects of 3.5 GHz (a frequency commonly used in 5G communication) RFR on neurobehavior, gut microbiota, and gut-brain axis metabolites in mice. The results showed that exposure to 3.5 GHz RFR at 50 W/m2 for 1 h over 35 d induced anxiety-like behaviour in mice, accompanied by NLRP3-dependent neuronal pyroptosis in CA3 region of the dorsal hippocampus. In addition, the microbial composition was widely divergent between the sham and RFR groups. 3.5 GHz RFR also caused changes in metabolites of feces, serum, and brain. The differential metabolites were mainly enriched in glycerophospholipid metabolism, tryptophan metabolism, and arginine biosynthesis. Further correlation analysis showed that gut microbiota dysbiosis was associated with differential metabolites. Based on the above results, we speculate that dysfunctional intestinal flora and metabolites may be involved in RFR-induced anxiety-like behaviour in mice through neuronal pyroptosis in the brain. The findings provide novel insights into the mechanism of 5G RFR-induced neurotoxicity.

Role of Low-Energy (<20 eV) Secondary Electrons in the Extraterrestrial Synthesis of Prebiotic Molecules.

We demonstrate for the first time that Galactic cosmic rays with energies as high as ∼1010 eV can trigger a cascade of low-energy (<20 eV) secondary electrons that could be a significant contributor to the interstellar synthesis of prebiotic molecules whose delivery by comets, meteorites, and interplanetary dust particles may have kick-started life on Earth. For the energetic processing of interstellar ice mantles inside dark, dense molecular clouds, we explore the relative importance of low-energy (<20 eV) secondary electrons-agents of radiation chemistry-and low-energy (<10 eV), nonionizing photons-instigators of photochemistry. Our calculations indicate fluxes of ∼102 electrons cm-2 s-1 for low-energy secondary electrons produced within interstellar ices due to attenuated Galactic cosmic-ray protons. Consequently, in certain star-forming regions where internal high-energy radiation sources produce ionization rates that are observed to be a thousand times greater than the typical interstellar Galactic ionization rate, the flux of low-energy secondary electrons should far exceed that of nonionizing photons. Because reaction cross sections can be several orders of magnitude larger for electrons than for photons, even in the absence of such enhancement, our calculations indicate that secondary low-energy (<20 eV) electrons are at least as significant as low-energy (<10 eV) nonionizing photons in the interstellar synthesis of prebiotic molecules. Most importantly, our results demonstrate the pressing need for explicitly incorporating low-energy electrons in current and future astrochemical simulations of cosmic ices. Such models are critically important for interpreting James Webb Space Telescope infrared measurements, which are currently being used to probe the origins of life by studying complex organic molecules found in ices near star-forming regions.

rTMS concurrent with cognitive training rewires AD brain by enhancing GM-WM functional connectivity: a preliminary study.

Repetitive transcranial magnetic stimulation (rTMS) and cognitive training for patients with Alzheimer's disease (AD) can change functional connectivity (FC) within gray matter (GM). However, the role of white matter (WM) and changes of GM-WM FC under these therapies are still unclear. To clarify this problem, we applied 40 Hz rTMS over angular gyrus (AG) concurrent with cognitive training to 15 mild-moderate AD patients and analyzed the resting-state functional magnetic resonance imaging before and after treatment. Through AG-based FC analysis, corona radiata and superior longitudinal fasciculus (SLF) were identified as activated WM tracts. Compared with the GM results with AG as seed, more GM regions were found with activated WM tracts as seeds. The averaged FC, fractional amplitude of low-frequency fluctuation (fALFF), and regional homogeneity (ReHo) of the above GM regions had stronger clinical correlations (r/P = 0.363/0.048 vs 0.299/0.108, 0.351/0.057 vs 0.267/0.153, 0.420/0.021 vs 0.408/0.025, for FC/fALFF/ReHo, respectively) and better classification performance to distinguish pre-/post-treatment groups (AUC = 0.91 vs 0.88, 0.65 vs 0.63, 0.87 vs 0.82, for FC/fALFF/ReHo, respectively). Our results indicated that rTMS concurrent with cognitive training could rewire brain network by enhancing GM-WM FC in AD, and corona radiata and SLF played an important role in this process.

Structural Insight into Polymerase Mechanism via a Chiral Center Generated with a Single Selenium Atom.

DNA synthesis catalyzed by DNA polymerase is essential for all life forms, and phosphodiester bond formation with phosphorus center inversion is a key step in this process. Herein, by using a single-selenium-atom-modified dNTP probe, we report a novel strategy to visualize the reaction stereochemistry and catalysis. We capture the before- and after-reaction states and provide explicit evidence of the center inversion and in-line attacking SN2 mechanism of DNA polymerization, while solving the diastereomer absolute configurations. Further, our kinetic and thermodynamic studies demonstrate that in the presence of Mg2+ ions (or Mn2+), the binding affinity (Km) and reaction selectivity (kcat/Km) of dGTPαSe-Rp were 51.1-fold (or 19.5-fold) stronger and 21.8-fold (or 11.3-fold) higher than those of dGTPαSe-Sp, respectively, indicating that the diastereomeric Se-Sp atom was quite disruptive of the binding and catalysis. Our findings reveal that the third metal ion is much more critical than the other two metal ions in both substrate recognition and bond formation, providing insights into how to better design the polymerase inhibitors and discover the therapeutics.

High-resolution single-photon imaging with physics-informed deep learning.

High-resolution single-photon imaging remains a big challenge due to the complex hardware manufacturing craft and noise disturbances. Here, we introduce deep learning into SPAD, enabling super-resolution single-photon imaging with enhancement of bit depth and imaging quality. We first studied the complex photon flow model of SPAD electronics to accurately characterize multiple physical noise sources, and collected a real SPAD image dataset (64 × 32 pixels, 90 scenes, 10 different bit depths, 3 different illumination flux, 2790 images in total) to calibrate noise model parameters. With this physical noise model, we synthesized a large-scale realistic single-photon image dataset (image pairs of 5 different resolutions with maximum megapixels, 17250 scenes, 10 different bit depths, 3 different illumination flux, 2.6 million images in total) for subsequent network training. To tackle the severe super-resolution challenge of SPAD inputs with low bit depth, low resolution, and heavy noise, we further built a deep transformer network with a content-adaptive self-attention mechanism and gated fusion modules, which can dig global contextual features to remove multi-source noise and extract full-frequency details. We applied the technique in a series of experiments including microfluidic inspection, Fourier ptychography, and high-speed imaging. The experiments validate the technique's state-of-the-art super-resolution SPAD imaging performance.

Polydopamine modified cellulose nanocrystals (CNC) for efficient cellulase immobilization towards advanced bamboo fiber flexibility and tissue softness.

Herein, a green facile approach to improve the flexibility of unbleached bamboo kraft pulp (UBKP) via an immobilized enzyme technology is proposed. Polydopamine (PDA) acts as versatile modification and coating materials of cellulose nanocrystals (CNC) for assembling versatile bio-carriers (PDA@CNC). Cellulase biomacromolecules are efficiently immobilized on PDA@CNC to form cellulase@PDA@CNC nanocomposites. The relative enzyme activity, temperature/pH tolerance, and storage stability of cellulase were significantly improved after immobilization. The degree of polymerization treated UBKP decreased by 5.42 % (25 U/g pulp) compared to the control sample. The flexibility of treated fibers was 6.61 × 1014/(N·m2), which was 96.93 % higher (25 U/g) compared to the control and 3.88 times higher than that of the blank fibers. Cellulase@PDA@CNC performs excellent accessibility to fiber structure and induces high degree of fibrillation and hydrolysis of UBKP fibers, which contributes high softness of obtained tissue handsheets. The bio-carrier PDA@CNC within paper framework may further enhance tissue tensile strength. This study proposes a practical and environmentally friendly immobilization approach of cellulase@PDA@CNC for improving the hydrolysis efficiency and flexibility of UBKP fibers, which provides the possibility to maintain the strength of tissue paper while improving its softness, thus broadening the high-value application of immobilized enzyme technology in tissue production.

Identification and Determination of Impurities in a New Therapeutic Agent for Fatty Liver Disease.

Methyl 7,7'-dimethoxy-5'-(morpholinomethyl)-[4,4'-bibenzo[d][1,3] dioxole]-5-carboxylate methanesulfonate (IMM) is an innovative drug for the treatment of nonalcoholic fatty liver disease (NAFLD) owing to its high efficacy and low toxicity. In this study, five minor impurities (I, II, III, IV, and V) were identified and analyzed using spectroscopic evidence, chemical synthetic methods, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The impurities included hydrolysates and oxidation by-products extracted from both the drug in its final formulation and during synthesis. Toxicity prediction revealed potential carcinogenicity of impurity V containing an N-oxygen fragment. A reliable and selective HPLC method for the quantitative analysis of impurities I-IV and a sensitive HPLC-MS/MS method for potential genotoxic impurity V were developed and optimized. The methods were validated based on the International Council for Harmonization guidelines. Satisfactory linearity was obtained for the analytes over the range of 0.1-2.0 µg/mL for impurities I-IV and 0.3-30.0 ng/mL for impurity V, and in all cases, the fitting correlation coefficients exceeded 0.999. The obtained limits of detection values were 0.05 ng/mL and 0.005 µg/mL for impurity V and impurities I-IV, respectively. The precision and repeatability of the methods were less than 1.08% and 8.72% for each impurity. The recovery percentages of all impurities were in the range of 91.18%-111.27%, with the relative standard deviation of less than 3.69%. The greenness assessment of the HPLC method and the HPLC-MS/MS method were evaluated by using AGREE software with a score value of 0.72 and 0.68, respectively. The recommended procedures that were accurate, specific, and ecofriendly were applied to the existing active pharmaceutical ingredients of IMM, and they generated satisfactory results.

Conjoint analysis of transcriptome and metabolome profiles of normal captivity and arch soil free-range in Meishan pigs.

The hygiene hypothesis has been advanced as a potential explanation for the increasingly high levels of atopy and allergic disease in the general human population. In an effort to conduct a more detailed study of the link between immune activity and the hygiene hypothesis, Meishan pigs raised under normal captivity (NC) or arch soil free-range (ASF) conditions were selected as an experimental model system. Cytokine levels were found to differ significantly between these two groups consistent with a difference in cellular immune status. Integrated transcriptomic and metabolomic profiling of duodenal tissue samples from Meishan pigs were then performed, leading to the identification of differentially expressed genes (DEGs), differentially abundant metabolites (DAMs), and key pathways that were able to distinguish the NC and ASF groups. This approach led to the identification of 1,113 DEGs, as well as 577 and 372 DAMs in positive and negative ion modes, respectively. When an interaction network incorporating DEGs and metabolites associated with immune responsivity was constructed, it included factors such as 9-cis-Retinoic acid, (9Z,11E)-(13S)-13-Hydroxyoctadeca-9,11-dienoic acid and (10E,12Z)-(9S)-9-Hydroxyoctadeca-10,12-dienoic acid. Functional enrichment analyses confirmed that identified DEGs and DAMs were associated with immune-related pathways including the intestinal IgA production and PPAR signaling pathways. Together, these results offer new insight into the roles that particular genes and metabolites enriched in response to environmental stressors in free-range Meishan pigs may play in the regulation of cellular immunity, thus offering a foundation for future efforts to better understand the immunological mechanisms underlying the hygiene hypothesis.

Molecular Characteristics of Bovine Viral Diarrhea Virus Strains Isolated from Persistently Infected Cattle.

In this study, we reported the isolation, identification, and molecular characteristics of nine BVDV strains that were isolated from the serum of persistently infected cattle. The new strains were designated as BVDV TJ2101, TJ2102, TJ2103, TJ2104, TJ2105, TJ2106, TJ2107, TJ2108 and TJ2109. The TJ2102 and TJ2104 strains were found to be cytopathic BVDV, and the other strains were non-cytopathic BVDV. An alignment and phylogenetic analysis showed that the new isolates share 92.2-96.3% homology with the CP7 strain and, thus, were classified as the BVDV-1b subgenotype. A recombination analysis of the genome sequences showed that the new strains could be recombined by the major parent BVDV-1a NADL strain and the minor parent BVDV-1m SD-15 strain. Some genome variations or unique amino acid mutations were found in 5'-UTR, E0 and E2 of these new isolates. In addition, a potential linear B cell epitopes prediction showed that the potential linear B cell epitope at positions 56-61 is highly variable in BVDV-1b. In conclusion, the present study has identified nine strains of BVDV from persistently infected cattle in China. Further studies on the virulence and pathogenesis of these new strains are recommended.

[Application progress of hypercrosslinked porous organic polymers in cartridge-based solid phase extraction].

Hypercrosslinked porous organic polymers (HCPs), a novel type of porous materials synthesized via the Friedel-Crafts reaction, are widely used in gas storage, heterogeneous catalysis, chromatographic separation, and organic pollutant capture. HCPs have the advantages of a wide monomer source, low cost, mild synthesis conditions, and easy functionalization. In recent years, HCPs have shown great application potential in solid phase extraction. Given their high specific surface area, excellent adsorption properties, diverse chemical structures, and easy chemical modification, HCPs have been successfully applied to the extraction of different types of analytes with efficient extraction performance. Based on the chemical structure of HCPs, their target analytes, and the adsorption mechanism, HCPs can be classified as hydrophobic, hydrophilic, and ionic species. Hydrophobic HCPs are usually constructed as extended conjugated structures by overcrosslinking aromatic compounds as monomers. Common monomers include ferrocene, triphenylamine, triphenylphosphine, etc. This type of HCPs shows good adsorption effects on nonpolar analytes such as benzuron herbicides and phthalates through strong π-π and hydrophobic interactions. Hydrophilic HCPs are prepared by introducing polar monomers or crosslinking agents, or by modifying polar functional groups. This type of adsorbent is commonly used to extract polar analytes such as nitroimidazole, chlorophenol, tetracycline, etc. In addition to hydrophobic forces, polar interactions, such as hydrogen-bonding and dipole-dipole interactions, also occur between the adsorbent and analyte. Ionic HCPs are mixed-mode solid phase extraction materials formed by introducing ionic functional groups into the polymer. Mixed-mode adsorbents usually have a dual reversed-phase/ion-exchange retention mechanism, which helps control the retention behavior of the adsorbent by adjusting the elution strength of the eluting solvent. In addition, the extraction mode can be switched by controlling the pH of the sample solution and eluting solvent. In this manner, matrix interferences can be removed while the target analytes are enriched. Ionic HCPs present a unique advantage in the extraction of acid-base drugs in water. The combination of new HCP extraction materials with modern analytical techniques, such as chromatography and mass spectrometry, has been widely used in environmental monitoring, food safety, and biochemical analyses. In this review, the characteristics and synthesis methods of HCPs are briefly introduced, and the application progress of different types of HCPs in cartridge-based solid phase extraction is described. Finally, the future outlook of HCP applications is discussed.

Image-free single-pixel object detection.

Recently developed image-free sensing techniques have achieved remarkable performance in various vision tasks. However, existing image-free methods still cannot simultaneously obtain the category, location, and size information of all objects. In this Letter, we report a novel image-free single-pixel object detection (SPOD) technique. SPOD enables efficient and robust multi-object detection directly from a small number of measurements, eliminating the requirement for complicated image reconstruction. Different from the conventional full-size pattern sampling method, the reported small-size optimized pattern sampling method achieves higher image-free sensing accuracy with fewer pattern parameters (∼1 order of magnitude). Moreover, instead of simply stacking CNN layers, we design the SPOD network based on the transformer architecture. It can better model global features and reinforce the network's attention to the targets in the scene, thus improving the object detection performance. We demonstrate the effectiveness of SPOD on the Voc dataset, which achieves a detection accuracy of 82.41% mAP at a sampling rate of 5% with a refresh rate of 63 f.p.s.

Phase retrieval via nonlocal complex-domain sparsity.

Phase retrieval is indispensable for a number of coherent imaging systems. Owing to limited exposure, it is a challenge for traditional phase retrieval algorithms to reconstruct fine details in the presence of noise. In this Letter, we report an iterative framework for noise-robust phase retrieval with high fidelity. In the framework, we investigate nonlocal structural sparsity in the complex domain by low-rank regularization, which effectively suppresses artifacts caused by measurement noise. The joint optimization of sparsity regularization and data fidelity with forward models enables satisfying detail recovery. To further improve computational efficiency, we develop an adaptive iteration strategy that automatically adjusts matching frequency. The effectiveness of the reported technique has been validated for coherent diffraction imaging and Fourier ptychography, with ≈7 dB higher peak SNR (PSNR) on average, compared with conventional alternating projection reconstruction.

Machine learning- and structure-based discovery of a novel chemotype as FXR agonists for potential treatment of nonalcoholic fatty liver disease.

Farnesoid X receptor (FXR) is a promising target for drug discovery against nonalcoholic fatty liver disease (NAFLD). However, no FXR agonist has been approved for NAFLD so far. The R & D of FXR agonists are somewhat hindered by the lack of effective and safe chemotypes. To this end, we developed a multi-stage computational workflow to screen the Specs and ChemDiv chemical library for FXR agonists, which consisted of machine learning (ML)-based classifiers, shape-based and electrostatic-based models, a FRED-based molecular docking protocol, an ADMET prediction protocol and substructure search. As a result, we identified a novel chemotype that has never been reported before, with compound XJ02862 (ChemDiv ID: Y020-6413) as the representative. By designing an asymmetric synthesis strategy, we were able to prepare four isomers of compound XJ02862. Interestingly, one of the isomers, 2-((S)-1-((2S,4R)-2-methyl-4-(phenylamino)-3,4-dihydroquinolin-1(2H)-yl)-1-oxopropan-2-yl)hexahydro-1H-isoindole-1,3(2H)-dione (XJ02862-S2), showed potent FXR agonistic activity in HEK293T cells. The molecular docking, molecular dynamics simulations and site-directed mutagenesis suggested the hydrogen bond between compound XJ02862-S2 and HIS294 of FXR is essential for ligand binding. We further demonstrated that compound XJ02862-S2 had no agonistic effect on TGR5. Further biological experiments have shown that compound XJ02862-S2 could ameliorate hypercholesterolemia, hepatic steatosis, hyperglycemia, insulin resistance (IR) in high-fat-diet induced obese (DIO) mice. In term of molecular mechanism, compound XJ02862-S2 regulates the expression of FXR downstream genes involved in lipogenesis, cholesterol transport and bile acid biosynthesis and transport. Taken together, we have discovered a novel chemotype as potent FXR agonists for NAFLD by computational modeling, chemical synthesis and biological evaluation.

Novel PORCN inhibitor WHN-88 targets Wnt/ß-catenin pathway and prevents the growth of Wnt-driven cancers.

Wnt/ß-catenin signaling pathway is a classical and crucial oncogenic pathway in many carcinomas, and Porcupine (PORCN) is an O-acyltransferase, which is indispensable and highly specific for catalyzing palmitoylation of Wnt ligands and facilitating their secretion and biofunction. Targeting PORCN provides a promising approach to specifically cure Wnt-driven cancers from the root. In this study, we designed series of pyridonyl acetamide compounds, and discovered a novel PORCN inhibitor WHN-88 with a unique di-iodinated pyridone structural fragment, which is significantly different from the reported inhibitors. We demonstrated that WHN-88 effectively abolished palmitoylation of Wnt ligands and prevented their secretion and the subsequent Wnt/ß-catenin signaling transduction. Further experiments showed that, at well-tolerated doses, WHN-88 remarkably suppressed the spontaneous occurrence and growth of MMTV-Wnt1 murine breast tumors. Consistently, WHN-88 also notably restrained the progress of xenografted Wnt-driven human tumors, including PA-1 teratocarcinoma with high autocrine Wnt signaling and Aspc-1 pancreatic carcinoma with Wnt-sensitizing RNF43 mutation. Additionally, we disclosed that WHN-88 inhibited cancer cell stemness obviously. Together, we verified WHN-88 is a novel PORCN inhibitor with potent efficacy against the Wnt-driven cancers. Our findings enriched the structural types of PORCN inhibitors, and facilitated the development and application of PORCN inhibiting therapy in clinic.

Vasoplegic syndrome in patients undergoing heart transplantation.

Objectives: To summarize the risk factors, onset time, and treatment of vasoplegic syndrome in patients undergoing heart transplantation. Methods: The PubMed, OVID, CNKI, VIP, and WANFANG databases were searched using the terms "vasoplegic syndrome," "vasoplegia," "vasodilatory shock," and "heart transplant*," to identify eligible studies. Data on patient characteristics, vasoplegic syndrome manifestation, perioperative management, and clinical outcomes were extracted and analyzed. Results: Nine studies enrolling 12 patients (aged from 7 to 69 years) were included. Nine (75%) patients had nonischemic cardiomyopathy, and three (25%) patients had ischemic cardiomyopathy. The onset time of vasoplegic syndrome varied from intraoperatively to 2 weeks postoperatively. Nine (75%) patients developed various complications. All patients were insensitive to vasoactive agents. Conclusions: Vasoplegic syndrome can occur at any time during the perioperative period of heart tranplantation, especially after the discontinuation of bypass. Methylene blue, angiotensin II, ascorbic acid, and hydroxocobalamin have been used to treat refractory vasoplegic syndrome.

Inhalable vaccine of bacterial culture supernatant extract mediates protection against fatal pulmonary anthrax.

Pulmonary anthrax is the most fatal clinical form of anthrax and currently available injectable vaccines do not provide adequate protection against it. Hence, next-generation vaccines that effectively induce immunity against pulmonary anthrax are urgently needed. In the present study, we prepared an attenuated and low protease activity Bacillus anthracis strain A16R-5.1 by deleting five of its extracellular protease activity-associated genes and its lef gene through the CRISPR-Cas9 genome editing system. This mutant strain was then used to formulate a lethal toxin (LeTx)-free culture supernatant extract (CSE) anthrax vaccine, of which half was protective antigen (PA). We generated liquid, powder, and powder reconstituted formulations that could be delivered by aerosolized intratracheal inoculation. All of them induced strong humoral, cellular, and mucosal immune responses. The vaccines also produced LeTx neutralizing antibodies and conferred full protection against the lethal aerosol challenges of B. anthracis Pasteur II spores in mice. Compared to the recombinant PA vaccine, the CSE anthrax vaccine with equal PA content provided superior immunoprotection against pulmonary anthrax. The preceding results suggest that the CSE anthrax vaccine developed herein is suitable and scalable for use in inhalational immunization against pulmonary anthrax.

Structural optimization and biological evaluation of 1-adamantylcarbonyl-4-phenylpiperazine derivatives as FXR agonists for NAFLD.

Farnesoid X receptor (FXR) is an attractive target for drug discovery against non-alcoholic fatty liver disease (NAFLD). We previously reported an orally active, new-chemotype FXR agonist XJ034 by ensemble learning-driven drug discovery. However, its FXR agonistic activity and the efficacy in vivo remain to be improved. In this study, we designed and synthesized 52 derivatives, and preliminarily evaluated their FXR transactivation activity in HEK293T cells at the concentration of 10 µM. 12 FXR agonists were superior or comparable to compound XJ034, two of which showed over 9-fold activity of compound XJ034, and were as potent as OCA. The molecular docking and molecular dynamics simulations implied an additional hydrogen bond with TYR383 is involved in FXR transactivation for both compounds. According to EC50 determined by the confirmatory transactivation assay, we selected adamantan-1-yl(4-(2-amino-5-chlorophenyl)piperazin-1-yl)methanone (10a, EC50: 1.05 µM) as our lead compound. Interestingly, compound 10a had no agonistic effect on TGR5 or PPAR, and no cytotoxicity to HepG2 cells. In vivo bioassays with high-fat-diet induced C57BL/6J obese (DIO) mice have shown that compound 10a (100 mg/kg) is more effective than compound XJ034 (200 mg/kg) in improving hyperlipidemia, hepatic steatosis and insulin resistance. We also observed that compound 10a down-regulated the expression of genes involved in liver inflammation in vivo, implying its potential to treat hepatic inflammation. In summary, the present data have proved that our strategy for structural optimization is effective, and compound 10a is a promising lead compound with improved efficacy for NAFLD.

Loss-of-function CFTR p.G970D missense mutation might cause congenital bilateral absence of the vas deferens and be associated with impaired spermatogenesis.

Congenital bilateral absence of the vas deferens (CBAVD) is observed in 1%-2% of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator (CFTR) mutations. CFTR is one of the most well-known genes related to male fertility. The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia (NOA). CFTR mutations are highly polymorphic and have established ethnic specificity. Compared with F508Del in Caucasians, the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis. However, whether p.G970D participates in male infertility remains unknown. Herein, a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA. Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation (4.1%, 5/122), excluding polymorphic sites. Furthermore, we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients. The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells. In spermatocyte GC-2(spd)ts (GC2) Cftr p.G965D cells, RNA splicing variants were detected and CFTR expression decreased, which may contribute to the phenotypes associated with impaired spermatogenesis. Thus, this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.

Loss-of-function CFTR p.G970D missense mutation might cause congenital bilateral absence of the vas deferens and be associated with impaired spermatogenesis / 亚洲男科学杂志(英文版)

Congenital bilateral absence of the vas deferens (CBAVD) is observed in 1%-2% of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator (CFTR) mutations. CFTR is one of the most well-known genes related to male fertility. The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia (NOA). CFTR mutations are highly polymorphic and have established ethnic specificity. Compared with F508Del in Caucasians, the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis. However, whether p.G970D participates in male infertility remains unknown. Herein, a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA. Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation (4.1%, 5/122), excluding polymorphic sites. Furthermore, we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients. The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells. In spermatocyte GC-2(spd)ts (GC2) Cftr p.G965D cells, RNA splicing variants were detected and CFTR expression decreased, which may contribute to the phenotypes associated with impaired spermatogenesis. Thus, this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.

Characterization of the VP2 and NS1 genes from canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) in Northern China.

Canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) cause severe disease in young animals, pups, and kittens. CPV-2 evolved from FPV by altering the species-specific binding of the viral capsid to the host receptor, i.e., the transferrin receptor (TfR), and CPV-2 genetic variants have been identified by specific VP2 amino acid residues (297, 426). Early studies focused on the main capsid protein VP2; however, there have been limited studies on the non-structural protein NS1. In this study, we identified the genetic variants of clinical samples in dogs and cats in northern China during 2019-2020. The genetic characterization and phylogenetic analyses of VP2 and NS1 gene were also conducted. The results revealed that the CPV-2c was identified as the major genetic variant. One new CPV-2b and two CPV-2c strains were collected from cats. Four mutation sites (60, 630, 443, and 545 amino acid residues) were located in the functional domains of the NS1 protein. The phylogenetic analysis of VP2 and NS1 genes showed that they were clustered by geographical regions and genotypes. The gene mutation rate of CPV-2 was increasing in recent years, resulting in a complex pattern of gene evolution in terms of host preference, geographical selection, and new genetic variants. This study emphasizes that continuous molecular epidemiological surveillance is required to understand the genetic diversity of FPV and CPV-2 strains.