Monoclonal antibody targeting a novel linear epitope on nucleoprotein confers pan-reactivity to influenza A virus.

Nucleoprotein (NP) functions crucially in the replicative cycle of influenza A virus (IAV) via forming the ribonucleoprotein complex together with PB2, PB1, and PA proteins. As its high conservation, NP ranks one of the hot targets for design of universal diagnostic reagents and antiviral drugs for IAV. Here, we report an anti-NP murine monoclonal antibody (mAb) 5F10 prepared from traditional lymphocyte hybridoma technique with the immunogen of a clade 2.3.4.4 H5N1 subtype avian influenza virus. The specificity of mAb 5F10 to NP protein was confirmed by immunofluorescence assay and western blotting, and the mAb 5F10 could be used in immunoprecipitation and immunohistochemistry assays. Importantly, mAb 5F10 possessed broad-spectrum reactivity against H1~H11 subtypes of avian influenza viruses, including various HA clades of H5Nx subtype. In addition, mAb 5F10 also showed good affinity with H1N1 and H3N2 subtype influenza viruses of swine and human origin. Furthermore, the recognized antigenic epitope of mAb 5F10 was identified to consist of the conserved amino acid motif 81EHPSA85 in the second flexible loop region of NP protein through screening the phage display peptide library. Collectively, the mAb 5F10 which recognizes the novel universal NP linear B-cell epitope of IAV with diverse origins and subtypes will be a powerful tool for NP protein-based structural, functional, and mechanistic studies, as well as the development of detection methods and universal vaccines for IAV. KEY POINTS: • A broad-spectrum mAb against various subtypes and sources of IAV was developed • The mAb possessed good reactivity in IFA, western blot, IP, and IHC assays • The mAb targeted a novel conserved linear B-cell epitope involving 81EHPSA85 on NP protein.

Zoonotic Threat of G4 Genotype Eurasian Avian-Like Swine Influenza A(H1N1) Viruses, China, 2020.

We investigated genetic and biologic characteristics of 2 Eurasian avian-like H1N1 swine influenza viruses from pigs in China that belong to the predominant G4 genotype. One swine isolate exhibited strikingly great homology to contemporaneous human Eurasian avian-like H1N1 isolates, preferential binding to the human-type receptor, and vigorous replication in mice without adaptation.

One-year clinical efficacy evaluation of selective corneal wavefront aberration-guided FS-LASIK correction in patients with high myopia.

AIM: To evaluate the effectiveness, safety, predictability, precision and changes of higher-order aberrations (HOAs) on corneal front surface of selective corneal wavefront aberration-guided femtosecond laser-assisted in situ keratomileusis (CW-FS-LASIK) in patients with high myopia 1-year postoperatively. METHODS: Totally 74 eyes of 37 patients with high myopia or myopic astigmatism in both the eyes who underwent the CW-FS-LASIK procedure in Xi'an Gaoxin Hospital from January 2021 to June 2021 were included. The changes of uncorrected distance visual acuity (UDVA), best corrected visual acuity (BCVA), spherical equivalent refraction (SER), astigmatism, HOAs and Strehl ratio (SR) on the anterior surface of the cornea after 1y of the surgery were analyzed. RESULTS: At postoperative 1y, the UDVA (logMAR) of 74 eyes (100%) reached 0 or better, including 0 in 8 eyes (10.81%), -0.1 in 45 eyes (60.81%), and -0.2 in 21 eyes (28.38%). The effectiveness index was 1.29±0.134. There was no decrease in postoperative BCVA compared with preoperative BCVA in all patients. Postoperative BCVA was the same in 44 eyes (59.46%) as preoperative BCVA, increased by 1 line in 23 eyes (31.08%) and increased by 2 lines in 7 eyes (9.46%) compared with preoperative BCVA. The safety index was 1.11±0.159. The estimated corrected SER before surgery was (-7.76±1.21) D, and the actual corrected SER was (-7.83±1.25) D (Y=0.9811X+0.2156, R2=0.9084). There was a high correlation between the estimated corrected SER and the actual corrected SER. The postoperative SER in 74 eyes (100%) was within ±0.75 D. The postoperative astigmatism of all was within -0.75 D to 0. Root mean square (RMS) HOAs of spherical aberration and SR within 5 mm of the corneal front surface were all increased compared with those before operation (P<0.01). The total coma, horizontal coma and vertical coma were all decreased compared with those before operation (P<0.01). There was no statistically significant difference in horizontal trefoil and vertical trefoil compared with preoperative ones (P>0.05). CONCLUSION: Selective CW-FS-LASIK for correction of high myopia is effective, safe, predictive, and accurate. For patients with preoperative RMS HOAs over 0.25 defocus equivalent, postoperative coma aberration can be significantly reduced, and SR value can be increased, thus corneal imaging quality can be improved.

Changes in corneal biomechanics and posterior corneal surface elevation after FS-LASIK.

AIM: To investigate the changes in corneal biomechanics and posterior corneal surface elevation after femtosecond laser-assisted in situ keratomileusis (FS-LASIK). METHODS: Totally 197 eyes of 100 patients who underwent the FS-LASIK from April 2022 to November 2022 were included. They were divided into three groups according to the ratio of residual corneal stroma thickness/corneal thickness (RCST/CT): Group I (50%≤RCST/CT<55%, 63 eyes of 32 patients), Group II (55%≤RCST/CT<60%, 67 eyes of 34 patients), and Group III (RCST/CT≥60%, 67 eyes of 34 patients). The intraocular pressure (IOP), corneal compensated IOP (IOPcc), corneal hysteresis (CH) and corneal resistance factor (CRF) were measured immediately, 1, and 3mo postoperatively by ocular response analyzer (ORA) and the posterior elevation difference (PED) was measured by Pentacam. RESULTS: After operation, IOP, CH, CRF, and PED were statistically different among the three groups (F=12.99, 31.148, 23.998, all P<0.0001). There was no statistically significant difference in IOPcc among the three groups (F=0.603, P>0.05). The IOP, IOPcc, CH, and CRF were statistical changed after surgery (F=699.635, 104.125, 308.474, 640.145, all P<0.0001). The PED of Group I was significantly higher than that of Group II (P<0.05), and Group II was significantly higher than that of Group III (P<0.05). The PED value of 3mo after surgery decreased in each group compared with 1mo after surgery, but there was no statistical difference (Group I: t=0.82, P=0.41; Group II: t=0.17, P=0.87; Group III: t=1.35, P=0.18). The correlation analysis of corneal biomechanical parameter changes with PED at 1mo and 3mo after surgery showed that ΔIOP, ΔIOPcc, ΔCH, and ΔCRF were not correlated with PED value in three groups (P>0.05). CONCLUSION: The smaller the RCST/CT, the greater effect on corneal biomechanics and posterior surface elevation. There is no correlation between changes in corneal biomechanics and posterior corneal surface elevation in the range of RCST/CT≥50%.

Multipronged Micelles-Hydrogel for Targeted and Prolonged Drug Delivery in Chronic Wound Infections.

Chronic diabetic wounds are a growing threat globally. Many aspects contribute to its deterioration, including bacterial infection, unbalanced microenvironment, dysfunction of cell repair, etc. In this work, we designed a multipronged micelles-hydrogel platform loaded with curcumin and rifampicin (CRMs-hydrogel) for bacteria-infected chronic wound treatment. The curcumin- and rifampicin-loaded micelles (CRMs) exhibited both MMP9-responsive and epidermal growth factor receptor (EGFR)-targeting abilities. On the one hand, drugs could be released from micelles due to responsive disassembly by MMP9, a matrix metalloproteinase overexpressed in a chronic wound environment; on the other hand, CRMs showed specific targeting to EGFR on epithelial cells and fibroblasts and therefore increased intracellular drug delivery. The thermosensitive CRMs-hydrogel could form strong adhesion with the wound area and served as a suitable matrix for sustained release of CRMs directly at the wound bed, with excellent intracellular and extracellular bacterial elimination efficiency and wound healing promotion capability. We found that a single dose of CRMs-hydrogel achieved 99% antibacterial rate at the MRSA-infected diabetic wound, which effectively reduced inflammatory response and promoted the neovascularization and re-epithelialization process, with nearly half reduction of the skin barrier regeneration period. Collectively, our thermosensitive, MMP9-responsive, and targeted micelles-hydrogel nanoplatform is promising for chronic wound treatment.

A new species of Rhinogobius (Pisces: Gobiidae), with analyses of its DNA barcode.

The genus Rhinogobius Gill 1859 is widely distributed in fresh waters along the Western Pacific coast of tropical and temperate Asia. A new species, Rhinogobius maxillivirgatus, is described from Anhui Province in Eastern China. This species can be differentiated from all congeners by a combination of the following characters: up to 6 longitudinal brown to black stripes along the side of the body; pectoral-fin rays modally 14; predorsal scale series 5-9; lateral scale series 28-30; transverse scale series 6-7; branchiostegal membrane with about 20 red round spots in males; and 2 black oblique stripes parallel along the upper jaw on the anterior portion of the cheek. Analyzing sequences of cytochrome c oxidase subunit I revealed that the new species is closely related to, but distinct, from Rhinogobius wuyanlingensis.