A novel XNA-based Luminex assay to detect UBA1 somatic mutations associated with VEXAS syndrome.

Objectives: Patients with VEXAS syndrome carry mutations of UBA1 gene coding for the E1 enzyme. The three most frequent mutations are p.M41T(122T > C), p.M41V (c.121A > G), and p.M41L (c.121A > C) in codon 41 of exon 3. Currently, sanger sequencing was mainly used to detect these mutations, which has low sensitivity and low throughput. There is a need of high sensitivity, simple and high throughput method to characterize patients with VEXAS syndrome. Methods: Based on our proprietary XNA technology, we have developed a QClamp® Plex platform to detect eight mutations in a single reaction using the Luminex xMap technology. The assay sensitivity, specificity and precision were subsequently evaluated. Furthermore, the reference interval and clinical sensitivity/specificity were estimated using clinical healthy/positive DNA samples and the sanger sequencing method was used for comparison. Results: With spiking synthetic mutant DNA in wildtype GM24385 cell line DNA, this assay can detect UBA1 mutations with a detection limit of variant allele frequency (VAF) at 0.1-5%. Our assay shows 100% concordance with Sanger sequencing results when used for analyzing 15 positive and 19 negative clinical samples. Conclusions: The QClamps® Plex UBA1 Mutation Detection Assay is a quicker, simpler, and more sensitive assay that can accurately detect the UBA1 mutations even at early stages with low mutation frequency.

A Stochastic Model Based on Optimal Satellite Subset Selection Strategy for Smartphone Pseudorange Relative Positioning.

In order to overcome the limitations of traditional stochastic models for smartphones, we introduce a double-difference code pseudorange residual (DDPR)-dependent stochastic model based on an optimal satellite subset, with the goal of mitigating the constraints imposed by the quality of GNSS observations in smartphones on the accuracy and reliability of phone-based GNSS positioning. In our methodology, the satellite selection process involved considering the integrated carrier-to-noise density ratio (C/N0) index of both the reference station and the smartphone, enabling us to construct a satellite subset characterized by superior observation quality. Furthermore, by leveraging the optimal subset of satellites and incorporating the C/N0-dependent stochastic model, we could determine the approximate location of the terminal through pseudorange differential positioning. Subsequently, we estimated the DDPRs for all satellites and utilized these values as prior information to build a stochastic model of the observations. Our findings indicate that in occluded environments, the DDPR-dependent stochastic model significantly enhances positioning accuracy for both the Huawei Mate40 and P40 terminals compared to the C/N0-dependent model. Numerically, the improvements in the north (N), east (E), and up (U) directions were approximately 30%, 32%, and 34% for the Mate40, and 26%, 33%, and 24% for the P40 terminal. This suggests that the proposed DDPR-dependent stochastic model effectively identifies and mitigates large gross error signals caused by multipath and non-line-of-sight (NLOS) signals, thereby assigning lower weights to these problematic observations and ultimately enhancing positioning accuracy. Moreover, the weighting method involves minimal computations and is straightforward to implement, making it particularly suitable for GNSS positioning applications on smartphones in complex urban environments.

Effects of cluster nursing interventions on the prevention of pressure ulcers in intensive care units patients: A meta-analysis.

A meta-analysis was conducted to comprehensively evaluate the impact of cluster nursing interventions on the prevention of pressure ulcers (PUs) in intensive care unit (ICU) patients. Computer searches were performed in databases including Embase, Google Scholar, Cochrane Library, PubMed, Wanfang and China National Knowledge Infrastructure for randomized controlled trials (RCTs) implementing cluster nursing interventions for PUs prevention in ICU patients, with the search period covering the database inception to November 2023. Two researchers independently screened the literature, extracted data and conducted quality assessments. Stata 17.0 software was employed for data analysis. Overall, 17 RCTs involving 1463 ICU patients were included. The analysis showed that compared with conventional nursing, cluster nursing interventions significantly reduced the incidence of PUs (odds ratio: 0.24, 95% confidence intervals [CI]: 0.17-0.34, p < 0.001) and also significantly improved the levels of anxiety (standardized mean difference [SMD]: -1.39, 95% CI: -1.57 to 1.22, p < 0.001) and depression (SMD: -1.64, 95% CI: -2.02 to 1.26, p < 0.001) in ICU patients. This study indicates that the application of cluster nursing interventions in ICU patients can effectively reduce the incidence of PUs, as well as improve patients' anxiety and depression levels, thereby enhancing their quality of life, which is worth clinical promotion and application.

A Highly Sensitive XNA-Based RT-qPCR Assay for the Identification of ALK, RET, and ROS1 Fusions in Lung Cancer.

Lung cancer is often triggered by genetic alterations that result in the expression of oncogenic tyrosine kinases. Specifically, ALK, RET, and ROS1 chimeric receptor tyrosine kinases are observed in approximately 5-7%, 1-2%, and 1-2% of NSCLC patients, respectively. The presence of these fusion genes determines the response to tyrosine kinase inhibitors. Thus, accurate detection of these gene fusions is essential in cancer research and precision oncology. To address this need, we have developed a multiplexed RT-qPCR assay using xeno nucleic acid (XNA) molecular clamping technology to detect lung cancer fusions. This assay can quantitatively detect thirteen ALK, seven ROS1, and seven RET gene fusions in FFPE samples. The sensitivity of the assay was established at a limit of detection of 50 copies of the synthetic template. Our assay has successfully identified all fusion transcripts using 50 ng of RNA from both reference FFPE samples and cell lines. After validation, a total of 77 lung cancer patient FFPE samples were tested, demonstrating the effectiveness of the XNA-based fusion gene assay with clinical samples. Importantly, this assay is adaptable to highly degraded RNA samples with low input amounts. Future steps involve expanding the testing to include a broader range of clinical samples as well as cell-free RNAs to further validate its applicability and reliability.

Targeting and Microenvironment-Activated Nanoreactor for Diabetic Chronic Wound Healing via Multienzyme Cascade Reactions.

The development of cell-like nanoreactors with the ability to initiate biocatalytic cascades under special conditions holds tremendous potential for therapeutic applications. Herein, conformationally gated nanoreactors that respond to the acidic microenvironment of infected diabetic wounds were developed by cucur[8]bituril (CB[8])-based supramolecular assembly. The bioinspired nanoreactors exhibit not only self-regulated permeability and selectivity to control internal enzyme activities by substance exchange but also distinct binding specificities toward Gram-positive and Gram-negative bacteria via noncovalent modification with different ligands. The encapsulation of glucose oxidase (GOx), Fe3O4 nanozyme, and l-arginine (l-Arg) into the nanocarriers enables intelligent activation of multienzyme cascade reactions upon glucose (Glu) uptake to produce gluconic acid (GA) and hydrogen peroxide (H2O2), which is further converted into highly toxic hydroxyl radicals (·OH) for selective antibacterial activity. Moreover, acidic H2O2 promotes the oxidization of l-Arg, leading to the release of nitric oxide (NO). Consequently, this nanoreactor provides a multifunctional and synergistic platform for diabetic chronic wound healing by combining enzyme dynamic therapy with NO gas therapy to combat bacterial infections and inflammation under high blood Glu levels.

Near-infrared imaging of hepatocellular carcinoma and its medicinal treatment with a γ-glutamyl transpeptidase-monitoring fluorescence probe.

Herein, the Near-infrared imaging of hepatocellular carcinoma (HCC) and its medicinal treatment was achieved with a γ-glutamyl transpeptidase (GGT)-monitoring fluorescence probe KYZ-GGT which consisted of the typical recognition group γ-glutamyl and the structurally modified signal reporting group hemicyanine-thioxanthene. Compared with the recently reported probes, KYZ-GGT suggested practical and steady capability for monitoring the GGT level in the cellular, xenograft, induced as well as medicinal treatment HCC models. It realized the mitochondrial targeting intracellular imaging to reflect the GGT dynamics in the induction or medicinal treatment of HCC. In the xenograft and induced model mice with multiple factors, KYZ-GGT showed stable performance for visualizing the HCC status. In the medicinal treatment of the long-period-induced HCC model mice verified by the serum indexes and histopathological analysis, KYZ-GGT successfully imaged the medicinal treatment process of HCC with two marketed drugs (Sorafenib and Lenvatinib) respectively, with an applicative penetration depth. The information here was meaningful for investigating effective medicinal strategies for overcoming HCC.

Imaging Investigation of Hepatocellular Carcinoma Progress via Monitoring γ-Glutamyltranspeptidase Level with a Near-Infrared Fluorescence/Photoacoustic Bimodal Probe.

Hepatocellular carcinoma (HCC) is one of the main principal causes of cancer death, and the late definite diagnosis limits therapeutic approaches in time. The early diagnosis of HCC is essential, and the previous investigations on the biomarkers inferred that the γ-glutamyltranspeptidase (GGT) level could indicate the HCC process. Herein, a near-infrared fluorescence/photoacoustic (NIRF/PA) bimodal probe, CySO3-GGT, was developed for monitoring the GGT level and thus to image the HCC process. After the in-solution tests, the bimodal response was convinced. The various HCC processes were imaged by CySO3-GGT at the cellular level. Then, the CCl4-induced HCC (both induction and treatment) and the subcutaneous and orthotopic xenograft mice models were selected. All throughout the tests, CySO3-GGT achieved NIRF and PA bimodal imaging of the HCC process. In particular, CySO3-GGT could effectively realize 3D imaging of the HCC nodule by visualizing the boundary between the tumor and the normal tissue. The information here might offer significant guidance for the dynamic monitoring of HCC in the near future.

Elastic Stable Intramedullary Nailing Improves Fracture Healing and Mobility in Children with Femoral Fractures.

Objective: To explore the application value and safety of elastic stable intramedullary nailing (ESIN) in pediatric femoral fractures (FFs), providing more reliable safety for the treatment of FFs in the future. Methods: This study selected 60 cases of pediatric FFs who completed fracture treatment in our hospital between March 2014 and January 2023, with 32 cases undergoing ESIN fixation included in the research group (RG) and another 28 cases receiving plate internal fixation assigned to the control group (CG). The operative time (OT), intraoperative blood loss (IBL), incision length, fracture healing time, fixator removal time, weight-bearing time, and hospital length of stay (HLOS) of the two groups were counted, and the pain of the children was evaluated by the Visual Analogue Scale (VAS). The clinical efficacy and complication rate were recorded, and the hip and knee functions before and after treatment were evaluated by the Hospital for Special Surgery (HSS) score. After the completion of treatment, the child's family was surveyed about their satisfaction with the treatment. Results: The research group had less OT, IBL, and incision length, as well as shorter fracture healing time, fixator removal time, weight-bearing time, and HLOS than the control group (P < .05), with markedly lower VAS scores at 12h-48h postoperatively (P < .05). In addition, the research group demonstrated an obviously higher overall response rate (96.88%) and a lower complication rate (15.63%) than the control group (P < .05). Furthermore, HSS scores and treatment satisfaction were higher in the research group than in the control group (P < .05). Conclusions: ESIN is a highly effective treatment for pediatric femoral fractures, leading to accelerated fracture healing, improved mobility, and exhibiting high clinical application value.

Response of Plant and Soil N, P, and N:P Stoichiometry to N Addition in China: A Meta-Analysis.

Nitrogen (N) and phosphorus (P) are key elements essential for plant growth and development. Due to fertilizer application, rapid urbanization, and fossil fuel combustion, nitrogen deposition has reached relatively high levels in China. However, there is still uncertainty regarding the response of N:P stoichiometry in plants and soil to N deposition across different ecosystems. Therefore, a meta-analysis was conducted using 845 observations from 75 studies to evaluate the response of plant and soil N and P concentrations and N to P ratios across various ecosystems to N addition. The analysis revealed that N concentration and N:P stoichiometry in plants and soil increased under N addition, while P concentration in plants and soil decreased on average. Furthermore, the magnitude of these responses was related to the N input rate and experimental duration. Finally, the effects of N addition on N concentration, P concentration, and N:P in terrestrial ecosystems would alter their allocation patterns, depending on relevant climate factors such as mean annual temperature and mean annual precipitation. This study highlights the ecological impact of N addition on the biogeochemical cycling of major elements (N and P) in terrestrial ecosystems in China. These findings are necessary for improving our understanding of the characteristics of plant ecological stoichiometry and helping to plan measures for increasing N deposition.

Dichromatic "Breather Molecules" in a Mode-Locked Fiber Laser.

Bound states of solitons ("molecules") occur in various settings, playing an important role in the operation of fiber lasers, optical emulation, encoding, and communications. Soliton interactions are generally related to breathing dynamics in nonlinear dissipative systems, and maintain potential applications in spectroscopy. In the present work, dichromatic breather molecules (DBMs) are created in a synchronized mode-locked fiber laser. Real-time delay-shifting interference spectra are measured to display the temporal evolution of the DBMs, that cannot be observed by means of the usual real-time spectroscopy. As a result, robust out-of-phase vibrations are found as a typical intrinsic mode of DBMs. The same bound states are produced numerically in the framework of a model combining equations for the population inversion in the mode-locked laser and cross-phase-modulation-coupled complex Ginzburg-Landau equations for amplitudes of the optical fields in the fiber segments of the laser cavity. The results demonstrate that the Q-switching instability induces the onset of breathing oscillations. The findings offer new possibilities for the design of various regimes of the operation of ultrafast lasers.

Nrf2/PINK1-mediated mitophagy induction alleviates sodium fluoride-induced hepatic injury by improving mitochondrial function, oxidative stress, and inflammation.

Mitophagy has distinct functions, which can lead to either protection or damage of tissues. Though current evidence indicated that NaF triggers mitophagy, the role and regulation of mitophagy in sodium fluoride (NaF)-induced liver injury still remain unclear. Therefore, we exployed the cell and mouse models and confirmed that NaF treatment activates mitophagy. Knocking down PTEN-induced putative kinase protein 1 (PINK1) expression attenuated mitophagy and increased the degree of mitochondrial impairment, oxidative stress, and apoptosis in NaF-treated HepG2 cells. In vivo experiments indicated that PINK1 deficiency weakened NaF-induced mitophagy. Moreover, PINK1-deficient mices aggravated NaF-induced hepatic mitochondrial injury, oxidative stress, and inflammation in livers, evidenced by the increased number of abnormal mitochondria, decreased adenosine triphosphate (ATP) and glutathione (GSH) levels, elevated reactive oxygen species (ROS) and malondialdehyde (MDA) content, enhanced hepatic macrophage infiltration and inflammatory cytokine levels. Notably, NaF exposure activated Nrf2 signaling both in vitro and in vivo. Nrf2 siRNA transfection blocked the upregulation of PINK1 expression and the induction of mitophagy in NaF-treated HepG2 cells. Also, ML385 (Nrf2 inhibitor) partially blocked the upregulation of PINK1 expression caused by NaF in mice livers. To sum up, the present study provided the demonstration that Nrf2/PINK1-mediated mitophagy activation offers a hepatoprotective effect by inhibiting NaF-induced mitochondrial dysfunction, oxidative stress, and inflammation.

The Common Neural Mechanism of Somatic Symptoms of Depression and Anxiety Disorders: A Resting-State Functional Magnetic Resonance Imaging Study.

INTRODUCTION: Somatic symptoms often occur as a manifestation of depression and anxiety. The subgenual anterior cingulate cortex (sgACC) has been shown to be closely related to both depression and anxiety and plays an important role in somatic symptoms. However, little is known regarding whether the abnormal function of the sgACC contributes to the common somatic symptoms of depression and anxiety. METHODS: Resting-state functional connectivity (RSFC) analysis based on the seed of the sgACC was investigated in 23 major depressive disorder (MDD) patients with somatic symptoms, 20 generalized anxiety disorder (GAD) patients with somatic symptoms, and 22 demographically matched healthy controls (HCs). The severity of depression, anxiety, and somatic symptoms was assessed using the Hamilton Depression Rating Scale (HAMD), Hamilton Anxiety Rating Scale (HAMA), and the 15-item somatic symptom severity scale from the Patient Health Questionnaire (PHQ-15), respectively. An analysis of covariance analysis (ANCOVA) was conducted to determine RSFC alterations among GAD, MDD, and HC groups with age, gender, and head motion as covariates. Correlation analyses were conducted between the RSFC of the sgACC and PHQ-15. RESULTS: The significantly different RSFC of right sgACC among the three groups was found in right STG, left cerebellum, and right postcentral. Post hoc analysis indicated that both MDD and GAD patients showed a decreased RSFC between the right sgACC and right STG than HCs, and both were negatively correlated with the PHQ-15 scores. CONCLUSION: The abnormally decreased RSFC of the sgACC and STG may be the underlying common mechanisms of depression and anxiety combined with somatic symptoms.

Identification of a novel cell cycle-related risk signature predicting prognosis in patients with pancreatic adenocarcinoma.

BACKGROUND: Growing evidence have indicated that cell cycle-related genes (CRGs) play an essential role in the progression of pancreatic adenocarcinoma (PAAD). Nevertheless, the application of CRGs in estimating the prognosis of PAAD patients is still lacking. This study aimed to establish a risk signature based on CRGs that can predict patients' overall survival for PAAD. METHODS: The expression and corresponding clinical data of PAAD patients from The Cancer Genome Atlas database and 200 cell cycle-related genes from the MSigDB were used for the generation and validation of the signature. LASSO Cox regression was applied to build the prediction model. The diagnostic value of signature was evaluated by receiver operating characteristic curves. Univariate and multivariate regression was used to construct the nomogram providing the clinicians a useful tool. RESULTS: A total of 103 CRGs were identified. Seven genes (RBM14, SMAD3, CENPA, KIF23, NUSAP1, INCENP, SMC4) with non-zero coefficients in LASSO analysis were used to construct the prognostic signature. The 7-gene signature significantly stratified patients into high- and low-risk groups in terms of overall survival, and the area under the receiver operating characteristic curve of 5-year survival reached 0.749. Multivariate analysis showed that the signature is an independent prognostic factor. We then mapped a nomogram to predict 1-, 3-, and 5-year survival for PAAD patients. The calibration curves indicated that the model was reliable. Finally, we discovered that TP53 and KRAS mutated most frequently in low and high-risk groups, respectively. CONCLUSION: Our findings suggested that the seven genes identified in this study are valuable prognostic predictors for patients with PAAD. These findings provided us with a novel insight that it is useful for understanding cell cycle mechanisms and for identifying patients with PAAD with poor prognosis.

Systematic review and meta-analysis of the incidence rates of adverse events after digestive endoscopy in children.

Background: With the widespread use of digestive endoscopy in children, a variety of adverse events (AEs) have occurred after digestive endoscopy. However, there are notable differences in the incidence of adverse reactions in digestive endoscopy in children at present, which makes it difficult to assess the safety of digestive endoscopy in children. Methods: Studies related to digestive endoscopy in children were screened from January 2005 to October 2021 from PubMed, Web of Science, Spring, CNKI, and Science Direct databases. RevMan5.3 and Stata were employed to carry out meta-analysis on the incidence of adverse respiratory events, myoclonus, abdominal pain, fever, bleeding, chest pain, sore throat, vomiting, and delayed capsule discharge after digestive endoscopy in children. The article quality was evaluated by the Agency for Healthcare Research and Quality (AHRQ). The chi-square test and I2 were adopted to test literature heterogeneity, and the article publication bias was assessed by displaying an inverted funnel plot as a funnel plot. Results: In all, 15 articles were included, involving a total of 27,770 children. In all, 15 articles were included, involving a total of 27,770 children. The risk ratio (RR) value of adverse respiratory events after digestive endoscopy in children was 1.31 [95% confidence interval (CI): 1.17 to 1.47, P<0.00001]; the odds ratio (OR) value of the incidence of myoclonus was 1.21 (95% CI: 1.01 to 1.46, P=0.04); the incidence of abdominal pain was 1.18 (95% CI: 1.11 to 1.27, P<0.00001); the incidence of fever was 1.09 (95% CI: 1.06 to 1.12, P<0.00001); the incidence of bleeding was 1.24 (95% CI: 0.94 to 1.64, P=0.13); the incidence of chest pain was 1.06 (95% CI: 1.03 to 1.09, P<0.0001); incidence of sore throat was 1.11 (95% CI: 1.05 to 1.18, P=0.0004); incidence of vomiting was 1.13 (95% CI: 1.06 to 1.21, P=0.0001); and the incidence of delayed capsule expulsion was 1.18 (95% CI: 1.00 to 1.40, P=0.05). Discussion: The incidence of AEs after digestive endoscopy in children was low, which can be used in the diagnosis and therapy of digestive system diseases in children.

Safety and immunogenicity of COVID-19 vaccination in patients with hepatocellular carcinoma (CHESS-NMCID 2101): A multicenter prospective study.

Data on safety and immunogenicity of coronavirus disease 2019 (COVID-19) vaccinations in hepatocellular carcinoma (HCC) patients are limited. In this multicenter prospective study, HCC patients received two doses of inactivated whole-virion COVID-19 vaccines. The safety and neutralizing antibody were monitored. Totally, 74 patients were enrolled from 10 centers in China, and 37 (50.0%), 25 (33.8%), and 12 (16.2%) received the CoronaVac, BBIBP-CorV, and WIBP-CorV, respectively. The vaccines were well tolerated, where pain at the injection site (6.8% [5/74]) and anorexia (2.7% [2/74]) were the most frequent local and systemic adverse events. The median level of neutralizing antibody was 13.5 (interquartile range [IQR]: 6.9-23.2) AU/ml at 45 (IQR: 19-72) days after the second dose of vaccinations, and 60.8% (45/74) of patients had positive neutralizing antibody. Additionally, lower γ-glutamyl transpeptidase level was related to positive neutralizing antibody (odds ratio = 1.022 [1.003-1.049], p = 0.049). In conclusion, this study found that inactivated COVID-19 vaccinations are safe and the immunogenicity is acceptable or hyporesponsive in patients with HCC. Given that the potential benefits may outweigh the risks and the continuing emergences of novel severe acute respiratory syndrome coronavirus 2 variants, we suggest HCC patients to be vaccinated against COVID-19. Future validation studies are warranted.

Evaluation of different combination of pam2CSK4, poly (I:C) and imiquimod enhance immune responses to H9N2 avian influenza antigen in dendritic cells and duck.

Current commercial H9 avian influenza viruses (AIVs) vaccines cannot provide satisfactory antibody titers and protective immunity against AIVs in duck. Toll like receptors (TLR) ligand as AIVs adjuvants can activate dendritic cells to improve immune responses in multiple animals, while the studies were absent in duck. Therefore, we investigated TLR ligands pam2CSK4, poly (I:C) and/or imiquimod enhance immune responses to inactivated H9N2 avian influenza antigen (H9N2 IAIV) in peripheral blood monocyte-derived dendritic cells (MoDCs) and duck. In vitro, we observed that transcription factor NF-κB, Th1/Th2 type cytokines (IFN-γ, IL-2 and IL-6) and the ability of catching H9N2 IAIV antigen were significantly up-regulated when H9N2 IAIV along with TLR ligands (pam2CSK4, poly (I:C) and imiquimod, alone or combination) in duck MoDCs. Also, the best enhancement effects were showed in combination of pam2CSK4, poly (I:C) and imiquimod group, whereas IFN-α showed no significant enhancement in all experimental groups. In vivo, the results demonstrated that the percentages of CD4+/ CD8+ T lymphocytes, the levels of Th1/Th2 type cytokines and H9N2 HI titers were significant enhanced in combination of pam2CSK4, poly (I:C) and imiquimod group. However, pam2CSK4 alone or combining with imiquimod showed no enhancement or additive effects on Th1 cytokines (IFN-γ and IL-2), Th2 cytokines (IL-6) and HI titers in Muscovy duck, respectively. Taken together, our results concluded that not all TLR ligands showed enhancement of immune responses to H9N2 IAIV in duck. The combination of poly (I:C), imiquimod and pam2CSK4 that can be an effectively adjuvant candidate for H9N2 AIVs inactivated vaccine in duck, which provide novel insights in explore waterfowl vaccine.

Current and Future Potential of Shellfish and Algae Mariculture Carbon Sinks in China.

Shellfish and algae mariculture make up an important part of the marine fishery carbon sink. Carbon sink research is necessary to ensure China achieves its goal of carbon neutrality. This study used the material quality assessment method to estimate the carbon sink capacity of shellfish and algae. Product value, carbon storage value, and oxygen release value were used to calculate the economic value of shellfish and algae carbon sequestration. The results showed that the annual average shellfish and algae carbon sink in China was 1.10 million tons from 2003 to 2019, of which shellfish accounted for 91.63%, wherein Crassostreagigas, Ruditapesphilippinarum, and Chlamysfarreri were the main contributors. The annual average economic value of China's shellfish and algae carbon sequestration was USD 71,303.56 million, and the product value was the main contributor, accounting for 99.11%. The carbon sink conversion ratios of shellfish and algae were 8.37% and 5.20%, respectively, thus making shellfish the aquaculture species with the strongest carbon sink capacity and the greatest carbon sink potential. The estimated growth rate in the shellfish and algae removable carbon sink was 33,900 tons/year in China, but this trend was uncertain. The capacity for carbon sequestration and exchange by aquaculture can be improved by expanding breeding space, promoting multi-level comprehensive breeding modes, and marine artificial upwelling projects.

Expression and Immunogenicity of Recombinant African Swine Fever Virus Proteins Using the Semliki Forest Virus.

African swine fever virus (ASFV) is a large DNA virus belonging to the Asfarviridae family that damages the immune system of pigs, resulting in the death or slaughter of millions of animals worldwide. Recent modern techniques in ASFV vaccination have highlighted the potential of viral replicon particles (RPs), which can efficiently express foreign proteins and induce robust cellular and humoral immune responses compared with the existing vaccines. In this study, we established a Semliki Forest virus (SFV) vector by producing replication-defective viral particles. This vector was used to deliver RPs expressing ASFV antigens. SFV-RPs expressing ASFV p32 (SFV-p32) and p54 (SFV-p54) were tested in baby hamster kidney (BHK-21) cells. Proteins expression was evaluated via western blotting and indirect immunofluorescence, while immunogenicity was evaluated in BALB/c mice. The resulting RPs exhibited high levels of protein expression and elicited robust humoral and cellular immune responses.

Interaction of HnRNP F with the guanine-rich segments in viral antigenomic RNA enhances porcine reproductive and respiratory syndrome virus-2 replication.

BACKGROUND: Heterogeneous nuclear ribonucleoprotein (HnRNP) F is a member of HnRNP family proteins that participate in splicing of cellular newly synthesized mRNAs by specifically recognizing tandem guanine-tracts (G-tracts) RNA sequences. Whether HnRNP F could recognize viral-derived tandem G-tracts and affect virus replication remain poorly defined. METHODS: The effect of HnRNP F on porcine reproductive and respiratory syndrome virus (PRRSV) propagation was evaluated by real-time PCR, western blotting, and plaque-forming unit assay. The association between HnRNP F and PRRSV guanine-rich segments (GRS) were analyzed by RNA pulldown and RNA immunoprecipitation. The expression pattern of HnRNP F was investigated by western blotting and nuclear and cytoplasmic fractionation. RESULTS: Knockdown of endogenous HnRNP F effectively blocks the synthesis of viral RNA and nucleocapsid (N) protein. Conversely, overexpression of porcine HnRNP F has the opposite effect. Moreover, RNA pulldown and RNA immunoprecipitation assays reveal that the qRMM1 and qRRM2 domains of HnRNP F recognize the GRS in PRRSV antigenomic RNA. Finally, HnRNP F is redistributed into the cytoplasm and forms a complex with guanine-quadruplex (G4) helicase DHX36 during PRRSV infection. CONCLUSIONS: These findings elucidate the potential functions of HnRNP F in regulating the proliferation of PRRSV and contribute to a better molecular understanding of host-PRRSV interactions.