Transcriptome and proteomic analysis of mpox virus F3L-expressing cells.

Background: Monkeypox or mpox virus (mpox) is a double-stranded DNA virus that poses a significant threat to global public health security. The F3 protein, encoded by mpox, is an apoenzyme believed to possess a double-stranded RNA-binding domain (dsRBD). However, limited research has been conducted on its function. In this study, we present data on the transcriptomics and proteomics of F3L-transfected HEK293T cells, aiming to enhance our comprehension of F3L. Methods: The gene expression profiles of pCAGGS-HA-F3L transfected HEK293T cells were analyzed using RNA-seq. Proteomics was used to identify and study proteins that interact with F3L. Real-time PCR was used to detect mRNA levels of several differentially expressed genes (DEGs) in HEK293T cells (or Vero cells) after the expression of F3 protein. Results: A total of 14,822 genes were obtained in cells by RNA-Seq and 1,672 DEGs were identified, including 1,156 up-regulated genes and 516 down-regulated genes. A total of 27 cellular proteins interacting with F3 proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and 19 cellular proteins with large differences in abundance ratios were considered to be candidate cellular proteins. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the DEGs were significantly enriched in immune-related pathways, including type I interferon signaling pathway, response to virus, RIG-I-like receptor signaling pathway, NOD-like receptor signaling pathway, etc. Moreover, some selected DEGs were further confirmed by real-time PCR and the results were consistent with the transcriptome data. Proteomics data show that cellular proteins interacting with F3 proteins are mainly related to RNA splicing and protein translation. Conclusions: Our analysis of transcriptomic and proteomic data showed that (1) F3L up-regulates the transcript levels of key genes in the innate immune signaling pathway, such as RIGI, MDA5, IRF5, IRF7, IRF9, ISG15, IFNA14, and elicits a broad spectrum of antiviral immune responses in the host. F3L also increases the expression of the FOS and JNK genes while decreasing the expression of TNFR2, these factors may ultimately induce apoptosis. (2) F3 protein interacts with host proteins involved in RNA splicing and protein translation, such as SNRNP70, POLR2H, HNRNPA1, DDX17, etc. The findings of this study shed light on the function of the F3 protein.

The complete mitochondrial genome and phylogenetic analysis of Polythlipta liquidalis Leech, 1889 (Crambidae: spilomelinae).

The complete mitochondrial genome of Polythlipta liquidalis Leech, 1889 was sequenced and annotated in this study, which was the first reported complete mitogenome of the genus Polythlipta. The mitogenome of P. liquidalis is 15,305 bp in length and was predicted to encode 37 typical mitochondrial genes including 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and one major non-coding A-T rich region. The maximum likelihood phylogenetic analysis based on the 13 PCGs was constructed, including P. liquidalis and 15 related Spilomelinae species, using Ostrinia furnacalis as the outgroup. The result showed that P. liquidalis is grouped with Sinomphisa plagialis. These data will serve as a molecular resource for species identification of P. liquidalis and become a valuable resource for a range of genetic, functional, evolutionary and comparative genomic studies on members of Spilomelinae.

[Protein expression, purification and mouse antiserum preparation of monkeypox virus A23R].

Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-ß-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.

Reduced binding activity of vaccine serum to omicron receptor-binding domain.

Coronavirus disease 2019 (COVID-19) vaccination regimens contribute to limiting the spread of severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2). However, the emergence and rapid transmission of the SARS-CoV-2 variant Omicron raise a concern about the efficacy of the current vaccination strategy. Here, we expressed monomeric and dimeric receptor-binding domains (RBDs) of the spike protein of prototype SARS-CoV-2 and Omicron variant in E. coli and investigated the reactivity of anti-sera from Chinese subjects immunized with SARS-CoV-2 vaccines to these recombinant RBDs. In 106 human blood samples collected from 91 participants from Jiangxi, China, 26 sera were identified to be positive for SARS-CoV-2 spike protein antibodies by lateral flow dipstick (LFD) assays, which were enriched in the ones collected from day 7 to 1 month post-boost (87.0%) compared to those harvested within 1 week post-boost (23.8%) (P < 0.0001). A higher positive ratio was observed in the child group (40.8%) than adults (13.6%) (P = 0.0073). ELISA results showed that the binding activity of anti-SARS-CoV-2 antibody-positive sera to Omicron RBDs dropped by 1.48- to 2.07-fold compared to its homogeneous recombinant RBDs. Thus, our data indicate that current SARS-CoV-2 vaccines provide restricted humoral protection against the Omicron variant.

Clinical and Structural Outcomes After Rotator Cuff Repair in Patients With Diabetes: A Meta-analysis.

BACKGROUND: The impact of diabetes on clinical and structural outcomes after rotator cuff repair remains controversial. PURPOSE/HYPOTHESIS: The purpose of this study was to compare clinical outcomes and retear rates after rotator cuff repair in patients with and without diabetes. Our hypotheses were that adequate control of diabetes would decrease the retear rate after rotator cuff repair and that patients with diabetes would have worse clinical outcomes. STUDY DESIGN: Systematic review; Level of evidence, 3. METHODS: The PubMed, Embase, and Cochrane Library databases were searched for studies comparing outcomes in patients with and without diabetes after full-thickness rotator cuff repair. Clinical outcome analysis included the Constant score, the American Shoulder and Elbow Surgeons (ASES) score, and the University of California-Los Angeles shoulder rating scale; we compared preoperative, postoperative, and change in functional scores from baseline to final follow-up among the included studies. The pooled relative risk was calculated using a random-effects model for retear rates. Clinical outcomes were also pooled using a random-effects model. RESULTS: Overall, 10 studies were included. Compared with patients without diabetes, patients with diabetes had a worse preoperative ASES score (P = .009) as well as worse postoperative Constant score (final follow-up range, 9-103 months; P = .0003). However, there was no significant difference in the absolute mean change in clinical outcomes between patients with and without diabetes. Diabetes was associated with a higher retear rate (19.3% in patients without diabetes vs 28.2% in patients with diabetes; P < .0001). The retear rate according to the severity of sustained hyperglycemia in the subgroup analysis was 14.6% in patients without diabetes, versus 22.7% in patients with well-controlled diabetes (<7.0% of preoperative serum HbA1c level; P = .12) and 40.0% in patients with uncontrolled diabetes (HbA1c level ≥7.0%; P < .00001). CONCLUSION: This meta-analysis suggests that diabetes mellitus is associated with an increased risk of retears after rotator cuff repair, and improved blood glucose control may reduce the risk of retears in patients with diabetes mellitus. Although effective glycemic control was associated with a decreased risk of retears in patients with diabetes, we could not prove causation because of potential bias and confounding in the included studies.

Enhancement of osteogenesis and biodegradation control by brushite coating on Mg-Nd-Zn-Zr alloy for mandibular bone repair.

To diminish incongruity between bone regeneration and biodegradation of implant magnesium alloy applied for mandibular bone repair, a brushite coating was deposited on a matrix of a Mg-Nd-Zn-Zr (hereafter, denoted as JDBM) alloy to control the degradation rate of the implant and enhance osteogenesis of the mandible bone. Both in vitro and in vivo evaluations were carried out in the present work. Viability and adhesion assays of rabbit bone marrow mesenchyal stem cells (rBM-MSCs) were applied to determine the biocompatibility of a brushite-coated JDBM alloy. Osteogenic gene expression was characterized by quantitative real-time polymerase chain reaction (RT-PCR). Brushite-coated JDBM screws were implanted into mandible bones of rabbits for 1, 4, and 7 months, respectively, using 316L stainless steel screws as a control group. In vivo biodegradation rate was determined by synchrotron radiation X-ray microtomography, and osteogenesis was observed and evaluated using Van Gieson's picric acid-fuchsin. Both the naked JDBM and brushite-coated JDBM samples revealed adequate biosafety and biocompatibility as bone repair substitutes. In vitro results showed that brushite-coated JDBM considerably induced osteogenic differentiation of rBM-MSCs. And in vivo experiments indicated that brushite-coated JDBM screws presented advantages in osteoconductivity and osteogenesis of mandible bone of rabbits. Degradation rate was suppressed at a lower level at the initial stage of implantation when new bone tissue formed. Brushite, which can enhance oeteogenesis and partly control the degradation rate of an implant, is an appropriate coating for JDBM alloys used for mandibular repair. The Mg-Nd-Zn-Zr alloy with brushite coating possesses great potential for clinical applications for mandibular repair.

[Effect of adrenomedullin on renal arteriole remodeling in spontaneous hypertensive rats].

OBJECTIVE: To investigate the effect of adrenomedullin (ADM) on renal arteriole remodeling and phosphorylation of extracellular signal-regulated protein kinases 1/2 (p-ERK1/2) in spontaneous hypertensive rats (SHR). METHODS: Male SHR (4 weeks old) were randomized into hypertensive group (SHR) and ADM-treated group (ADM) to receive subcutaneous saline and ADM injections (daily dose of 1.0 nmol/kg, 5 days a week), respectively, with age-matched Wistar-Kyota (WKY) rats as the blank control. The systolic blood pressure (SBP) was measured at the end of each week, and histological changes of the renal arterioles were observed using HE and Weigert staining; the expression of P-ERK1/2 in the arterioles was detected with immunohistochemistry and Western blotting. RESULTS: At 16 and 24 weeks of age, the rats in both SHR and ADM groups showed significantly higher SBP levels than WKY rats (P<0.05), and at 24 weeks, SBP was significantly lower in ADM group than in SHR group (P<0.05). The intima thickness/lumen diameter (IT/LD) ratio of the renal arterioles increased in both SHR and ADM groups at 16 and 24 weeks as compared with that of WKY rats (P<0.05), and for arterioles with an outer diameter <40 µm, the IT/LD ratio was significantly lower at 24 weeks in ADM group than in SHR group (P<0.05). The renal expression of p-ERK1/2, which increased significantly in SHR and ADM groups at 16 and 24 weeks (P<0.05), was significantly lower in ADM group than in SHR group at 24 weeks (P<0.05). CONCLUSIONS: Long-term ADM treatment can control SPB elevation in SHR rats and reduce renal arteriole remodeling by inhibiting the phosphorylation of ERK1/2.

[Prevalence of trachoma in rural primary school children in Tengzhou City of Shandong Province in China].

OBJECTIVE: To investigate the prevalence of trachoma and its risk factors in rural primary school children in Tengzhou City of Shandong Province. METHODS: In this cross-sectional population-based study, children aged 5 to 14 years old in primary school were randomly selected by a cluster sampling in which school shift was the sampling unit. Out of 2742 students, 2676 were eligible. The examination rate was 97.60%. All selected students were assessed for trachoma using the simplified grading scheme proposed by the World Health Organization (WHO). Statistical significance was calculated using Chi-square tests. RESULTS: Out of 2676 eligible children, 593 cases of active trachoma were found, the prevalence of trachoma was 22.16% (95% CI:20.59%-23.73%). In 1606 boys, the prevalence of active trachoma was 19.74%, compared with 25.79% for girls. Girls were affected by active trachoma higher than boys (25.79% versus 19.74%, P=0.000). There was no significant difference among different age groups in term of the prevalence of trachoma (P=0.052). The prevalence of trachomatous follicle (TF), trachomatous inflammation (TI), and trachomatous scarring (TS) was 5.68% (152/2676), 19.21% (514/2676), 0.56% (15/2676), respectively. TI was more prevalent in girls than in boys (22.90% versus 16.75%, P=0.000). CONCLUSIONS: Trachoma is still endemic in children of primary schools in Tengzhou rural areas. Some interventions including mass treatment with antibiotics, improvement of hygienic conditions, and improvement of primary eye care are needed.