Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system.

CRISPR/Cas9 technology applied to Plasmodium falciparum offers the potential to greatly improve gene editing, but such expectations including large DNA fragment knock-ins and sequential gene editing have remained unfulfilled. Here, we achieved a major advance in addressing this challenge, especially for creating large DNA fragment knock-ins and sequential editing, by modifying our suicide-rescue-based system that has already been demonstrated to be highly efficient for conventional gene editing. This improved approach was confirmed to mediate efficient knock-ins of DNA fragments up to 6.3 kb, to produce "marker-free" genetically engineered parasites and to show potential for sequential gene editing. This represents an important advancement in establishing platforms for large-scale genome editing, which might gain a better understanding of gene function for the most lethal cause of malaria and contribute to adjusting synthetic biology strategies to live parasite malaria vaccine development. Site-directed knock-in of large DNA fragments is highly efficient using suicide-rescue-based CRISPR/Cas9 system, and sequential gene insertion is feasible but further confirmation is still needed.

Prognostic nomogram for predicting lower extremity deep venous thrombosis in ruptured intracranial aneurysm patients who underwent endovascular treatment.

Background: Lower extremity deep vein thrombosis (DVT) is one of the major postoperative complications in patients with ruptured intracranial aneurysms (RIA) who underwent endovascular treatment (EVT). However, patient-specific predictive models are still lacking. This study aimed to construct and validate a nomogram model for estimating the risk of lower extremity DVT for RIA patients who underwent EVT. Methods: This cohort study enrolled 471 RIA patients who received EVT in our institution between 1 January 2020 to 4 February 2022. Perioperative information on participants is collected to develop and validate a nomogram for predicting lower extremity DVT in RIA patients after EVT. Predictive accuracy, discriminatory capability, and clinical effectiveness were evaluated by concordance index (C-index), calibration curves, and decision curve analysis. Result: Multivariate logistic regression analysis showed that age, albumin, D-dimer, GCS score, middle cerebral artery aneurysm, and delayed cerebral ischemia were independent predictors for lower extremity DVT. The nomogram for assessing individual risk of lower extremity DVT indicated good predictive accuracy in the primary cohort (c-index, 0.92) and the validation cohort (c-index, 0.85), with a wide threshold probability range (4-82%) and superior net benefit. Conclusion: The present study provided a reliable and convenient nomogram model developed with six optimal predictors to assess postoperative lower extremity DVT in RIA patients, which may benefit to strengthen the awareness of lower extremity DVT control and supply appropriate resources to forecast patients at high risk of RIA-related lower extremity DVT.

The role of neutrophils in corneal nerve regeneration.

BACKGROUND: To investigate the role of neutrophils in corneal nerve regeneration. METHODS: A mouse model simulating corneal nerve injury was established and samples from corneal scraping with and without neutrophil closure were collected. These samples were used for corneal nerve staining, ribonucleic acid sequencing, and bioinformatics. Differential expression analysis was used to perform enrichment analysis to identify any significant differences between these two groups. The differential genes were then intersected with neutrophil-associated genes and a protein-protein interaction network was constructed using the intersected genes. The immune infiltration between the two groups was examined along with the immune cell variation between the high and low gene expression groups. RESULTS: Neutrophil removal delays corneal epithelial and nerve regeneration. A total of 546 differential genes and 980 neutrophil-associated genes, with 27 genes common to both sets were obtained. Molecular Complex Detection analysis yielded five key genes, namely integrin subunit beta 2 (ITGB2), matrix metallopeptidase 9 (MMP9), epidermal growth factor (EGF), serpin family E member 1 (SERPINE1), and plasminogen activator urokinase receptor (PLAUR). Among these genes, ITGB2, SERPINE1, and PLAUR exhibited increased expression in the neutrophil-confined group, while MMP9 and EGF showed decreased expression, with MMP9 and EGF displaying a more significant difference. Immune infiltration was also observed between the two groups, revealing significant differences in the infiltration of M0 macrophages, activated mast cells, and neutrophils. Moreover, the neutrophil levels were lower in the groups with low MMP9 and EGF expressions and higher in the high-expression group. CONCLUSION: Neutrophil confinement might significantly affect the MMP9 and EGF expression levels. Strategies to inhibit MMP9 could potentially yield therapeutic benefits.

Boosting short-chain fatty acids production from co-fermentation of orange peel waste and waste activated sludge: Critical role of pH on fermentation steps and microbial function traits.

The anaerobic co-fermentation of orange peel waste (OPW) and waste activated sludge (WAS) for useful short-chain fatty acids (SCFAs) generation presents an environmentally friendly and efficient method for their disposal. This study amied to investigate the effects of pH regulation on OPW/WAS co-fermentation, and found that the alkaline pH regulation (pH 9) significantly enhanced the promotion of SCFAs (11843 ± 424 mg COD/L), with a high proportion of acetate (51%). Further analysis revealed that alkaline pH regulation facilitated solubilization, hydrolysis, and acidification while simultaneously inhibiting methanogenesis. Furthermore, the functional anaerobes, as well as the expressions of corresponding gene involved in SCFAs biosynthesis, were generally improved under alkaline pH regulation. Alkaline treatment might played a critical role in alleviating the toxicity of OPW, resulting in improving microbial metabolic activity. This work provided an effective strategy to recover biomass waste as high-value products, and insightful understanding of microbial traits during OPW/WAS co-fermentation.

Small Incision Lenticule Extraction (SMILE) and Laser in Situ Keratomileusis (LASIK) Used to Treat Myopia and Myopic Astigmatism: A Systematic Review and Meta-analysis of Randomized Clinical Trials.

PURPOSES: The purpose of this meta-analysis is to systematically compare the safety, efficacy, and predictability of small incision lenticule extraction (SMILE) and laser in situ keratomileusis (LASIK). METHODS: This study covered the data searched from the PubMed, the EMBASE and the Cochrane Library. The Cochrane Handbook was also referred to as evaluating the quality of the included studies. In addition, this meta-analysis was performed using Revman 5.4 software. RESULTS: A total of 11 randomized controlled trails (RCTs) were included. The proportion of eyes with refraction within ±0.5D was higher in LASIK group compared with SMILE group (RR, 0.91; 95% CI, 0.83 to 0.99; p = .04). The spherical aberration (SA) was smaller in SMILE group compared with LASIK group (RR, -0.12; 95% CI, -0.23 to -0.01; p = .04). There were no significant differences between two groups with regard to final mean refractive spherical equivalent (SE) (MD, -0.04; 95% CI, -0.12 to 0.03; p = .22), proportion of eyes losing one or more lines of corrected distance visual acuity (CDVA) (RR, 1.14; 95% CI, 0.58 to 2.27; p = .70), proportion of eyes with uncorrected distance visual acuity (UCVA) of 20/20 or better (RR, 0.99; 95% CI, 0.94 to 1.05; p = .71), postoperative mean logMAR UCVA (MD, 0.01; 95% CI, -0.00 to 0.03; p = .13), postoperative refraction within ±1.0D (RR, 1.00; 95% CI, 0.98 to 1.02; p = .60), postoperative astigmatism within ±0.25, 0.5 and 1.0D (RR, 0.80, 0.99, 1.00; 95% CI, 0.35 to 1.83, 0.94 to 1.05, 0.98 to 1.02; p = .60, 0.86, 0.87), postoperative higher order aberrations (HOAs) (RR, 0.00; 95% CI, -0.16 to 0.16; p = .99). CONCLUSION: For predictability, LASIK was superior to SMILE. There were comparably safety and efficacy for the correction of myopia and myopic astigmatism in SMILE and LASIK. SA was smaller after SMILE than after LASIK.

Microstent Implantation with Phacoemulsification Versus Phacoemulsification Alone for Patients with Open Angle Glaucoma: A Systematic Review and Meta-Analysis of Randomised Clinical Trials.

PURPOSES: The purpose of this meta-analysis is to systematically compare the IOP-lowering effect of different microstents combined with phacoemulsification versus phacoemulsification for patients with OAG and cataract. METHODS: This work was done through the data searched from PubMed, EMBASE, and the Cochrane Library. The Cochrane Handbook was also used to evaluate the quality of the included studies. In addition, this meta-analysis was performed using Revman 5.4 software. RESULTS: A total of 8 randomized controlled trials (RCTs) were included. Compared with phacoemulsification alone, microstent implantation with phacoemulsification resulted in significant reduction in the postoperative IOP (MD = -1.66, 95%CI: [-2.25 to -1.06]). Patients with medication free and patients with beyond 20% IOP reduction were significantly increased in the microstent implantation with phacoemulsification group compared with phacoemulsification alone group (RR = 1.54, 95%CI: [1.34 to 1.77]; RR = 1.34, 95%CI: [1.24 to 1.45]). CONCLUSION: Both microstent implantation with concurrent phacoemulsification and phacoemulsification alone result in a significant reduction in IOP. In terms of both reductions, microstent implantation with phacoemulsification significantly outperforms phacoemulsification alone.

Establishment of the New Zealand white rabbit animal model of fatty keratopathy associated with corneal neovascularization.

The term fatty keratopathy is used to describe the phenomenon of fat deposition caused by corneal neovascularization, which will severely affect the eye's beauty and vision. The purpose of this study was to establish a New Zealand white rabbit animal model of fatty keratopathy, that is, the establishment of an animal model of fatty keratopathy. The goal was achieved by the combination of a corneal neovascularization animal model and a hyperlipidemia animal model. Two groups were created according to the experimental sequence. The first group initially induced a corneal neovascularization pattern and later induced a hyperlipidemia pattern, and the second group followed the opposite sequence. The results of the two groups showed that all the significant crystalline deposits of the cornea were visible. So the animal models of fatty keratopathy were successfully established in both groups.

A method to alleviate false-positive results of the Elecsys HIV combi PT assay.

To explore the effects of urea dissociation on reducing false-positive results of  the Elecsys HIV combi PT assay. A retrospective analysis was used to evaluate the false-positive rate of the Elecsys HIV combi PT assay. Six false-positive sera, six positive sera and six sera from patients with early HIV infection were collected. Dissociation was performed using 1 mol/L, 2 mol/L, 4 mol/L, 6 mol/L, or 8 mol/L urea, and HIV screening assay were then detected to select the appropriate concentration of urea dissociation. Next, 55 false-positive sera and 15 sera from early HIV infection were used to verify the best concentration of urea to achieve dissociation. Retrospective analysis showed that the COI of the Elecsys HIV combi PT assay in false-positive sera ranged from 1.0 to 200.0, and approximately 97.01%(227/234) of false-positive sera were in the range of 1.0-15.0. The avidity index (AI) in positive and false-positive sera decreased as the urea dissociation concentration increased. When the dissociation concentration was 6 mol/L, the AI of false-positive serum was between 0.0234 and 0.2567, and the AI of early HIV infection sera was between 0.4325 and 0.5017. The difference in AI between false-positive and positive samples was significant. When negativity was defined as an AI of less than 0.3970, the sensitivity and specificity were 100.0% and 100.0%, respectively. Urea-mediated dissociation could significantly reduce the false-positive rate of the Elecsys HIV combi PT assay with a low COI. Our findings provided a reference for distinguishing positive and false-positive of the Elecsys HIV combi PT assay.

Public authority control strategy for opinion evolution in social networks.

This paper addresses the need to deal with and control public opinion and rumors. Existing strategies to control public opinion include degree, random, and adaptive bridge control strategies. In this paper, we use the HK model to present a public opinion control strategy based on public authority (PA). This means utilizing the influence of expert or high authority individuals whose opinions we control to obtain the optimum effect in the shortest time possible and thus reach a consensus of public opinion. Public authority (PA) is only influenced by individuals' attributes (age, economic status, and education level) and not their degree distribution; hence, in this paper, we assume that PA complies with two types of public authority distribution (normal and power-law). According to the proposed control strategy, our experiment is based on random, degree, and public authority control strategies in three different social networks (small-world, scale-free, and random) and we compare and analyze the strategies in terms of convergence time (T), final number of controlled agents (C), and comprehensive efficiency (E). We find that different network topologies and the distribution of the PA in the network can influence the final controlling effect. While the effect of PA strategy differs in different network topology structures, all structures achieve comprehensive efficiency with any kind of public authority distribution in any network. Our findings are consistent with several current sociological phenomena and show that in the process of public opinion/rumor control, considerable attention should be paid to high authority individuals.

Human infection with a novel, highly pathogenic avian influenza A (H5N6) virus: Virological and clinical findings.

BACKGROUND AND OBJECTIVES: Severe infection with avian influenza A (H5N6) virus in humans was identified first in 2014 in China. Before that, it was unknown or unclear if the disease or the pathogen affected people. This study illustrates the virological and clinical findings of a fatal H5N6 virus infection in a human patient. METHODS: We obtained and analyzed the clinical, epidemiological, and virological data from the patient. Reverse transcription polymerase chain reaction (RT-PCR), viral culture, and sequencing were conducted for determination of the causative pathogen. RESULTS: The patient, who presented with fever, severe pneumonia, leucopenia, and lymphopenia, developed septic shock and acute respiratory distress syndrome (ARDS), and died on day 10 after illness onset. A novel reassortant avian-origin influenza A (H5N6) virus was isolated from the throat swab or trachea aspirate of the patient. The virus was reassorted with the HA gene of clade 2.3.4.4 H5, the internal genes of clade 2.3.2.1 H5, and the NA gene of the H6N6 avian virus. The cleavage site of the HA gene contained multiple basic amino acids, indicating that the novel H5N6 virus was highly pathogenic in chicken. CONCLUSIONS: A novel, highly pathogenic avian influenza H5N6 virus with a backbone of H5N1 virus acquired from the NA gene from the H6N6 virus has been identified. It caused human infection resulting in severe respiratory disease.