Multiple mutations of SARS-CoV-2 Omicron BA.2 variant orchestrate its virological characteristics.

IMPORTANCE: Most studies investigating the characteristics of emerging SARS-CoV-2 variants have been focusing on mutations in the spike proteins that affect viral infectivity, fusogenicity, and pathogenicity. However, few studies have addressed how naturally occurring mutations in the non-spike regions of the SARS-CoV-2 genome impact virological properties. In this study, we proved that multiple SARS-CoV-2 Omicron BA.2 mutations, one in the spike protein and another downstream of the spike gene, orchestrally characterize this variant, shedding light on the importance of Omicron BA.2 mutations out of the spike protein.

A Live-Attenuated Zika Virus Vaccine with High Production Capacity Confers Effective Protection in Neonatal Mice.

Zika virus (ZIKV) infection during pregnancy has been linked to congenital abnormalities, such as microcephaly in infants. An efficacious vaccine is desirable for preventing the potential recurrence of ZIKV epidemic. Here, we report the generation of an attenuated ZIKV (rGZ02a) that has sharply decreased virulence in mice but grows to high titers in Vero cells, a widely approved cell line for manufacturing human vaccines. Compared to the wild-type ZIKV (GZ02) and a plasmid-launched rGZ02p, rGZ02a has 3 unique amino acid alterations in the envelope (E, S304F), nonstructural protein 1 (NS1, R103K), and NS5 (W637R). rGZ02a is more sensitive to type I interferon than GZ02 and rGZ02p, and causes no severe neurological disorders in either wild-type neonatal C57BL/6 mice or type I interferon receptor knockout (Ifnar1-/-) C57BL/6 mice. Immunization with rGZ02a elicits robust inhibitory antibody responses with a certain long-term durability. Neonates born to the immunized dams are effectively protected against ZIKV-caused neurological disorders and brain damage. rGZ02a as a booster vaccine greatly improves the protective immunity primed by Ad2-prME, an adenovirus-vectored vaccine expressing ZIKV prM and E proteins. Our results illustrate that rGZ02a-induced maternal immunity can be transferred to the neonates and confer effective protection. Hence, rGZ02a may be developed as an alternative live-attenuated vaccine and warrants further evaluation. IMPORTANCE Zika virus (ZIKV), a mosquito-borne flavivirus that has caused global outbreaks since 2013, is associated with severe neurological disorders, such as Guillian-Barré syndrome in adults and microcephaly in infants. The ZIKV epidemic has gradually subsided, but a safe and effective vaccine is still desirable to prevent its potential recurrence, especially in countries of endemicity with competent mosquito vectors. Here, we describe a novel live-attenuated ZIKV, rGZ02a, that carries 3 unique amino acid alterations compared to the wild-type GZ02 and a plasmid-launched rGZ02p. The growth capacity of rGZ02a is comparable to GZ02 in Vero cells, but the pathogenicity is significantly attenuated in two mice models. Immunization with rGZ02a elicits robust inhibitory antibody responses in the dams and effectively protects their offspring against ZIKV disease. Importantly, in a heterologous prime-boost regimen, rGZ02a effectively boosts the protective immunity primed by an adenovirus-vectored vaccine. Thus, rGZ02a is a promising candidate for a live-attenuated ZIKV vaccine.

Upregulating Lin28a Promotes Axon Regeneration in Adult Mice with Optic Nerve and Spinal Cord Injury.

Severed CNS axons fail to regenerate in adult mammals and there are no effective regenerative strategies to treat patients with CNS injuries. Several genes, including phosphatase and tensin homolog (PTEN) and Krüppel-like factors, regulate intrinsic growth capacity of mature neurons. The Lin28 gene is essential for cell development and pluripotency in worms and mammals. In this study, we evaluated the role of Lin28a in regulating regenerative capacity of diverse populations of CNS neurons in adult mammals. Using a neuron-specific Thy1 promoter, we generated transgenic mice that overexpress Lin28a protein in multiple populations of projection neurons, including corticospinal tracts and retinal ganglion cells. We demonstrate that upregulation of Lin28a in transgenic mice induces significant long distance regeneration of both corticospinal axons and the optic nerve in adult mice. Importantly, overexpression of Lin28a by post-injury treatment with adeno-associated virus type 2 (AAV2) vector stimulates dramatic regeneration of descending spinal tracts and optic nerve axons after lesions. Upregulation of Lin28a also enhances activity of the Akt signaling pathway in mature CNS neurons. Therefore, Lin28a is critical for regulating growth capacity of multiple CNS neurons and may become an important molecular target for treating CNS injuries.

Promoting Axon Regeneration in Adult CNS by Targeting Liver Kinase B1.

Liver kinase B1 (LKB1), a downstream effector of cyclic AMP (cAMP)/PKA and phosphatidylinositol 3-kinase (PI3K) pathways, is a determinant for migration and differentiation of many cells, but its role in CNS axon regeneration is unknown. Therefore, LKB1 was overexpressed in sensorimotor cortex of adult mice five days after mid-thoracic spinal cord injury, using an AAV2 vector. Regeneration of corticospinal axons was dramatically enhanced. Next, systemic injection of a mutant-AAV9 vector was used to upregulate LKB1 specifically in neurons. This promoted long-distance regeneration of injured corticospinal fibers into caudal spinal cord in adult mice and regrowth of descending serotonergic and tyrosine hydroxylase immunoreactive axons. Either intracortical or systemic viral delivery of LKB1 significantly improved recovery of locomotor functions in adult mice with spinal cord injury. Moreover, we demonstrated that LKB1 used AMPKα, NUAK1, and ERK as the downstream effectors in the cortex of adult mice. Thus, LKB1 may be a critical factor for enhancing the growth capacity of mature neurons and may be an important molecular target in the treatment of CNS injuries.

Establishing criteria for human mesenchymal stem cell potency.

This study sought to identify critical determinants of mesenchymal stem cell (MSC) potency using in vitro and in vivo attributes of cells isolated from the bone marrow of age- and sex-matched donors. Adherence to plastic was not indicative of potency, yet capacity for long-term expansion in vitro varied considerably between donors, allowing the grouping of MSCs from the donors into either those with high-growth capacity or low-growth capacity. Using this grouping strategy, high-growth capacity MSCs were smaller in size, had greater colony-forming efficiency, and had longer telomeres. Cell-surface biomarker analysis revealed that the International Society for Cellular Therapy (ISCT) criteria did not distinguish between high-growth capacity and low-growth capacity MSCs, whereas STRO-1 and platelet-derived growth factor receptor alpha were preferentially expressed on high-growth capacity MSCs. These cells also had the highest mean expression of the mRNA transcripts TWIST-1 and DERMO-1. Irrespective of these differences, both groups of donor MSCs produced similar levels of key growth factors and cytokines involved in tissue regeneration and were capable of multilineage differentiation. However, high-growth capacity MSCs produced approximately double the volume of mineralized tissue compared to low-growth capacity MSCs when assessed for ectopic bone-forming ability. The additional phenotypic criteria presented in this study when combined with the existing ISCT minimum criteria and working proposal will permit an improved assessment of MSC potency and provide a basis for establishing the quality of MSCs prior to their therapeutic application.

Use of fermented Chinese medicine residues as a feed additive and effects on growth performance, meat quality, and intestinal health of broilers.

Introduction: The purpose of this research was to investigate how dietary supplementation with fermented herbal residues (FCMR) affected birds' development capacity, quality of meat, gut barrier, and cecum microbiota. Methods: 540 cyan-shank partridge birds aged 47 days were chosen and divided into two groups of six replicates each and 45 birds for each replicate. The control group (CON) received a basal diet, while the trial group decreased a basic diet containing 5% FCMR. Results and discussion: The findings revealed that the addition of FCMR decreased FCR and increased ADG in broilers (P < 0.05). Adding FCMR increased steaming loss in broiler chicken breasts (p < 0.05). Supplementation with FCMR significantly enhanced VH/CD and VH in the bird's intestine (jejunum, duodenum, and ileum) (p < 0.05). In addition, the addition of FCMR significantly down-regulated mRNA expression of INF-γ, IL-6, IL-1ß, and TNF-α and up-regulated mRNA expression of ZO-1, Occludin, and Claudin (P < 0.05). Microbial 16S rDNA high-throughput sequencing study revealed that supplements with FCMR modified the cecum microbiota, and α-diversity analysis showed that supplementation with FCMR reduced the cecum bacterial abundance in broilers (P < 0.05). At the phylum level, the relative abundance of Spirochaetota increased considerably following FCMR supplementation (P < 0.05). The broiler cecum's close lot of Prevotellaceae_UCG-001 (P < 0.05), Desulfovibrio, Muribaculaceae, and Fusobacterium (p < 0.05) reduced when FCMR was supplemented. Supplementation with FCMR can promote growth capacity and maintain intestinal health in birds by enhancing gut barrier function and modulating the inflammatory response and microbial composition.

Virological Characteristics of Five SARS-CoV-2 Variants, Including Beta, Delta and Omicron BA.1, BA.2, BA.5.

SARS-CoV-2 variants of concern (VOCs) show increasing transmissibility and infectivity and induce substantial injuries to human health and the ecology. Therefore, it is vital to understand the related features for controlling infection. In this study, SARS-CoV-2 WIV04 (prototype) and five VOCs (Beta, Delta, Omicron BA.1, BA.2 and BA.5 variants) were inoculated in Vero cells to observe their growth activities. Apart from evaluating the environmental stability at different temperatures, residual virus titers and infectivity at different temperatures (4 °C, room temperature (RT) and 37 °C) were measured over 7 days. The experiment also assessed the infectivity for different incubation durations. The growth capacity assay suggested that the WIV04, Beta and Delta variants replicated efficiently in Vero cells compared with Omicron Variants, and BA.2 replicated more efficiently in Vero cells than BA.1 and BA.5. In addition, all variants exhibited longer survivals at 4 °C and could remain infectious after 7 days, compared to RT' survival after 5 days and at 37 °C after 1 day. The virus infection assay indicated that the Omicron variant had a weaker ability to infect cells compared to the WIV04, Beta and Delta strains, and a longer infection time was required for these strains, except for BA.2.

The trade-offs between resistance and resilience of forage stay robust with varied growth potentials under different soil water and salt stress.

Water shortage and soil salinization are important factors restricting crop production worldwide. To conduct accurate yield prediction and reasonable crop layout, more attention should be paid to the performances of crop resistance and resilience under water and salt stress and their trade-off relationships. Here, we set different water (full irrigation, W0; moderate deficit irrigation, W1; and severe deficit irrigation, W2) and salt (S0, S1, S2, S3, S4, S5, and S6, representing 0 ‰, 1 ‰, 2 ‰, 3 ‰, 4 ‰, 5 ‰, and 6 ‰ salt in soil) treatments. Together with relevant studies, we analyzed the performances of forage resistance (Rt) and resilience (Rs) and their relationships under varied water and salt stress. The results indicated that logarithmic Rt (lg(Rt), the same as lg(Rs)) and the distribution of lg(Rs) were affected by water and salt stress, however, the relationships of lg(Rs)-lg(Rt) stayed stable with the constant slopes (k) and declined intercepts (m) as stress intensified. The physiological mechanisms and trade-offs for fixed species remained robust while the growth potentials varied under stress, which were closely related to stomatal regulations. Forage with larger |k| was suitable for fully irrigated regions to achieve higher yields, while regions with detrimental water and salt conditions should select cultivars with smaller |k| to ensure production. This study laid the groundwork for the estimation of the perennial forage adaptation and stability, and the method of long-term yield prediction and cultivar management under soil water and salt stress.

Liver Kinase B1 Functions as a Regulator for Neural Development and a Therapeutic Target for Neural Repair.

The liver kinase B1 (LKB1), also known as serine/threonine kinase 11 (STK11) and Par-4 in C. elegans, has been identified as a master kinase of AMPKs and AMPK-related kinases. LKB1 plays a crucial role in cell growth, metabolism, polarity, and tumor suppression. By interacting with the downstream signals of SAD, NUAK, MARK, and other kinases, LKB1 is critical to regulating neuronal polarization and axon branching during development. It also regulates Schwann cell function and the myelination of peripheral axons. Regulating LKB1 activity has become an attractive strategy for repairing an injured nervous system. LKB1 upregulation enhances the regenerative capacity of adult CNS neurons and the recovery of locomotor function in adult rodents with CNS axon injury. Here, we update the major cellular and molecular mechanisms of LKB1 in regulating neuronal polarization and neural development, and the implications thereof for promoting neural repair, axon regeneration, and functional recovery in adult mammals.

Quercetin promotes motor and sensory function recovery following sciatic nerve-crush injury in C57BL/6J mice.

Injuries and diseases that occur in the nervous system are common and have few effective treatments. Previous studies have shown that quercetin has a therapeutic effect on nervous system injuries, but its potential effects on and mechanisms of action related to behavioral recovery and axonal regrowth have not been investigated. Here, we showed that quercetin administration promotes behavioral recovery following sciatic nerve-crush injury in mice. Long-term evaluation showed that mice administered 20 mg·kg-1·day-1 quercetin for 35 days had a greater sensorimotor recovery compared with all other treatment groups. The mechanisms behind these effects were further investigated, and quercetin was found to regulate the expression of genes involved in regeneration and trophic support. Moreover, quercetin increased cyclic adenosine monophosphate expression and downstream pathway activation, which directly leads to neuronal growth activation in peripheral axon regeneration. In addition, quercetin enhanced axon remyelination, motor nerve conduction velocity and plantar muscle function, indicating that the degree of distal portion hypotrophy during the peripheral axon regeneration process was reduced. These results suggest that quercetin accelerates functional recovery by up-regulating neuronal intrinsic growth capacity and postponing distal atrophy. Overall, quercetin triggered multiple effects to promote behavioral recovery following sciatic nerve-crush injury in mice.

PTEN inhibition and axon regeneration and neural repair.

The intrinsic growth ability of all the neurons declines during development although some may grow better than others. Numerous intracellular signaling proteins and transcription factors have been shown to regulate the intrinsic growth capacity in mature neurons. Among them, PI3 kinase/Akt pathway is important for controlling axon elongation. As a negative regulator of this pathway, the tumor suppressor phosphatase and tensin homolog (PTEN) appears critical to control the regenerative ability of young and adult neurons. This review will focus on recent research progress in axon regeneration and neural repair by PTEN inhibition and therapeutic potential of blocking this phosphatase for neurological disorders. Inhibition of PTEN by deletion in conditional knockout mice, knockdown by short-hairpin RNA, or blockade by pharmacological approaches, including administration of selective PTEN antagonist peptides, stimulates various degrees of axon regrowth in juvenile or adult rodents with central nervous system injuries. Importantly, post-injury PTEN suppression could enhance axonal growth and functional recovery in adult central nervous system after injury.

The relationships between relative growth rate, meristematic potential and compensatory growth of semiarid-land shrubs.

Inherent relative growth rate has been suggested as a major determinant of plant species' capacity to regrow and compensate for tissues lost to herbivores. We investigated: 1) the relationship between compensatory growth capacity and relative growth rate (RGR) in six semiarid-land shrubs following removal in winter or spring of 90% of the previous year's growth, 2) the influence of loss of buds on production of new growth and 3) the relationship between meristematic potential and the capacity to produce new growth in four of the six semiarid-land shrub species. Four-year-old plants growing under field conditions were used in the study. The species with the highest inherent growth rate, sagebrush [Artemisia tridentata ssp. vaseyana (Rydb.) Beetle], died following the severe clipping treatments. The other five species exactly compensated for lost tissues. Inherent growth rates and compensatory growth capacity of the shrubs were not correlated. Loss of 90% of the buds on the previous year's growth did not limit production of new growth. Instead, shrubs that lost buds produced more new growth biomass than the controls. The shrub species had significantly different meristematic potential. Curlleaf mountain mahogany (Cercocarpus ledifolius Nutt.) and serviceberry (Amelanchier alnifolia Nutt.) had the greatest and least number of buds and long shoots per plant, respectively. The number of long shoots produced following bud removal was positively correlated with new growth biomass, while the percentage of long shoots produced at the basal position on twigs was negatively correlated with new growth biomass. Our results suggest that inherent growth rate is not likely to influence production of new growth following browsing when resources for growth are not limiting. In contrast, the ability of a shrub to initiate long shoot growth is likely to influence production of new growth even when resources for growth are abundant.

The effect of systemic PTEN antagonist peptides on axon growth and functional recovery after spinal cord injury.

Knockout studies suggest that PTEN limits the regenerative capacities of CNS axons as a dominant antagonist of PI3 kinase, but the transgenic approach is not feasible for treating patients. Although application of bisperoxovanadium may block PTEN function, it is a general inhibitor of phosphotyrosine phosphatases and may target enzymes other than PTEN, causing side effects and preventing firm conclusions about PTEN inhibition on regulating neuronal growth. A pharmacological method to selectively suppress PTEN post-injury could be a valuable strategy for promoting CNS axon regeneration. We identified PTEN antagonist peptides (PAPs) by targeting PTEN critical functional domains and evaluated their efficacy for promoting axon growth. Four PAPs (PAP 1-4) bound to PTEN protein expressed in COS7 cells and blocked PTEN signaling in vivo. Subcutaneous administration of PAPs initiated two days after dorsal over-hemisection injury significantly stimulated growth of descending serotonergic fibers in the caudal spinal cord of adult mice. Systemic PAPs induce significant sprouting of corticospinal fibers in the rostral spinal cord and limited growth of corticospinal axons in the caudal spinal cord. More importantly, PAP treatment enhanced recovery of locomotor function in adult rodents with spinal cord injury. This study may facilitate development of effective therapeutic agents for CNS injuries.

The model of the hospital's growth capacity and its significance / 中华医院管理杂志

The paper reviews the evolution of the enterprise growth theory, pointing out that its shortcoming lies in its stress only on what the enterprise possesses while ignoring what the customers need. In view of the characteristics of the medical industry, the paper argues that similar problems can also be found in the model for assessing the integrated strength of the hospital. It proposes the use of the model of the hospital's growth capacity in assessing the future development of the hospital. The model is composed of three aspects: material resources, manpower, and brand competitiveness. The paper also discusses the practical values resulting from the use of the hospital's growth capacity as the model for assessing hospital development.