CRISPR/Cas9-mediated knockin of human factor IX into swine factor IX locus effectively alleviates bleeding in hemophilia B pigs

Hemophilia B is an X-linked recessive bleeding disorder caused by abnormalities in the coagulation factor IX gene. Without prophylactic treatment, patients experience frequent spontaneous bleeding episodes. Well-characterized animal models are valuable for determining the pathobiology of the disease and testing novel therapeutic innovations. Here, we generated a porcine model of hemophilia B using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. Moreover, we tested the possibility of hemophilia B therapy by gene insertion. Frequent spontaneous joint bleeding episodes that occurred in hemophilia B pigs allowed a thorough investigation of the pathological process of hemophilic arthropathy. In contrast to the hemophilia B pigs, which showed a severe bleeding tendency and joint damage, the transgenic pigs carrying human coagulation factor IX exhibited a partial improvement of bleeding. In summary, this study not only offers a translational hemophilia B model for exploring the pathological process of hemophilic arthropathy but also provides a possibility for the permanent correction of hemophilia in the future by genome editing in situ

Proton inhibition of unitary currents of vanilloid receptors

Protons, which are released during inflammation and injury, regulate many receptors and ion channels involved in pain transduction, including capsaicin channels (transient receptor potential vanilloid receptors 1). Whereas extracellular acidification both sensitizes and directly activates the channel, it also causes concomitant reduction of the unitary current amplitudes. Here, we investigate the mechanisms and molecular basis of this inhibitory effect of protons on channel conductance. Single-channel recordings showed that the unitary current amplitudes decreased with extracellular pH in a dose-dependent manner, consistent with a model in which protons bind to a site within the channel with an apparent pKa of ∼6. The inhibition was voltage dependent, ∼65% at −60 mV and 37% at +60 mV when pH was reduced from 7.4 to 5.5. The unitary current amplitudes reached saturation at [K+] ≥ 1 M, and notably the maximum amplitudes did not converge with different pHs, inconsistent with a blockade model based on surface charge screening or competitive inhibition of permeating ions. Mutagenesis experiments uncovered two acidic residues critical for proton inhibition, one located at the pore entrance and the other on the pore helix. Based on homology to the KcsA structure, the two acidic residues, along with another basic residue also on the pore helix, could form a triad interacting with each other through extensive hydrogen bonds and electrostatic contacts, suggesting that protons may mediate the interactions between the selectivity filter and pore helix, thereby altering the local structure in the filter region and consequently the conductance of the channel

Application of high performance liquid chromatography (HPLC) and erythrocyte parameters in the screening of hemoglobinopathy

Objective: To explore the application value of high performance liquid chromatography (HPLC) and erythrocyte parameters for screening of thalassemia in Shanghai. Methods: The hemoglobin of 1 029 samples from patients with suspected thalassemia and with suspected hemolytic anemia were analyzed using HPLC. Genetic analysis of the globin were performed simultaneously with GAP-PCR,reverse dot blot(RDB) and DNA sequencing. Results: Among 1029 samples, 591（57.43%） were diagnosed as having thalassemia and 14 (1.36%) having structural hemoglobin variants.Compared with results of genetic analysis,the sensitivity,specificity,positive predictive value and negative predictive value of HbA2 >3.9% by HPLC as cut-off value for diagnosing β-thalassemia were 95.84%, 96.87%, 95.40%, 97.20%,with AUC of 0.962. When HbA2≤2.1% by HPLC as the cutoff value combined with presence of rapid band were taken for diagnosing α-thalassemia moderate（HbH disease）, sensitivity, specificity, positive predictive value, negative predictive value were 96.32%, 52.60%, 100.00 %, with AUC of 0.990. If f HbA2 ≤3.2% by HPLC was taken as the cutoff value for diagnosing minor α-thalassemia or α-thalassemia trait, the corresponding indice were 97.76%, 53.11%, 24.0%, 99.4%,and AUC was 0.753. If MCV≤73.3 fl,MCH≤23.5 pg,MCHC≤324g/L were taken as the cutoff value alone for diagnosing thalassemia, sensitivity and specificity were 87.98%,77.51%, 93.73% and 69.86%, 89.20%,47.61%,respectively. Conclusions: Based on RBC parameters analysis,results of HPLC technology have a good consistence with the resluts of gene detection in the diagnosis of β-thalassemia, HbH disease and structural hemoglobin variant, while a low specificity and positive predictive value may occur in the diagnosis of minor α-thalassemia orα-thalassemia trait

Complement Factor B Mediates Ocular Angiogenesis through Regulating the VEGF Signaling Pathway

Complement factor B (CFB), a 95-kDa protein, is a crucial catalytic element of the alternative pathway (AP) of complement. After binding of CFB to C3b, activation of the AP depends on the proteolytic cleavage of CFB by factor D to generate the C3 convertase (C3bBb). The C3 convertase contains the catalytic subunit of CFB (Bb), the enzymatic site for the cleavage of a new molecule of C3 into C3b. In addition to its role in activating the AP, CFB has been implicated in pathological ocular neovascularization, a common feature of several blinding eye diseases, however, with somewhat conflicting results. The focus of this study was to investigate the direct impact of CFB on ocular neovascularization in a tightly controlled environment. Using mouse models of laser-induced choroidal neovascularization (CNV) and oxygen-induced retinopathy (OIR), our study demonstrated an increase in CFB expression during pathological angiogenesis. Results from several in vitro and ex vivo functionality assays indicated a promoting effect of CFB in angiogenesis. Mechanistically, CFB exerts this pro-angiogenic effect by mediating the vascular endothelial growth factor (VEGF) signaling pathway. In summary, we demonstrate compelling evidence for the role of CFB in driving ocular angiogenesis in a VEGF-dependent manner. This work provides a framework for a more in-depth exploration of CFB-mediated effects in ocular angiogenesis in the future

Coordinated State Analysis and Differential Regulation of Territorial Spatial Functions in Underdeveloped Regions: A Case Study of Gansu Province, China

The coordinated development of territorial spatial functions is the main manifestation of the sustainable utilization of territorial resources. Identifying spatial functions and their coordinated relationship has become an important guarantee for regional coordinated development, and is of great significance to the construction of a sustainable land development and protection pattern. This study constructs a classification and function scoring system for Production-Living-Ecological Spaces (PLES) by using land-use data from Gansu Province in 2000 and 2020, and revises the spatial functions of the land to supplement the deficiencies of existing related researches by combining socio-economic data (GDP, population density) and ecosystem service value data. In the aforementioned works, the Mechanical Equilibrium Model in physics is referenced to explore the coordinated state among territorial spatial functions. The results show that the high-value areas of production function are mainly distributed in the eastern and central areas of the Loess Plateau, with a strip-like distribution in the Hexi Corridor. The high-value areas of living function are consistent with that in the center of the city. The distribution of high-value areas of ecological function is in line with the topographic distribution pattern of mountainous areas in Gansu Province. The distribution pattern of the coordinated state of land space function in Gansu Province is relatively stable from 2000 to 2020. Simultaneously, the functional dominant area and the promotion area are identified according to the deviation of the coordinated degree of production-living-ecological function. After clarifying the functional characteristics of each county, the author proposes a differentiated regulation strategy of territorial spatial function

Virtual Reality Technology in Cognitive Rehabilitation Application: Bibliometric Analysis

BackgroundIn recent years, with the development of computer science and medical science, virtual reality (VR) technology has become a promising tool for improving cognitive function. Research on VR-based cognitive training has garnered increasing attention. ObjectiveThis study aimed to investigate the application status, research hot spots, and emerging trends of VR in cognitive rehabilitation over the past 20 years. MethodsArticles on VR-based cognitive rehabilitation from 2001 to 2021 were retrieved from the Web of Science Core Collection. CiteSpace software was used for the visual analysis of authors and countries or regions, and Scimago Graphica software was used for the geographic visualization of published countries or regions. Keywords were clustered using the gCLUTO software. ResultsA total of 1259 papers were included. In recent years, research on the application of VR in cognitive rehabilitation has been widely conducted, and the annual publication of relevant literature has shown a positive trend. The main research areas include neuroscience and neurology, psychology, computer science, and rehabilitation. The United States ranked first with 328 papers, and Italy ranked second with 140 papers. Giuseppe Riva, an Italian academic, was the most prolific author with 29 publications. The most frequently cited reference was “Using Reality to Characterize Episodic Memory Profiles in Amnestic Mild Cognitive Impairment and Alzheimer’s Disease: Influence of Active and Passive Encoding.” The most common keywords used by researchers include “virtual reality,” “cognition,” “rehabilitation,” “performance,” and “older adult.” The largest source of research funding is from the public sector in the United States. ConclusionsThe bibliometric analysis provided an overview of the application of VR in cognitive rehabilitation. VR-based cognitive rehabilitation can be integrated into multiple disciplines. We conclude that, in the context of the COVID-19 pandemic, the development of VR-based telerehabilitation is crucial, and there are still many problems that need to be addressed, such as the lack of consensus on treatment methods and the existence of safety hazards

Effect of Fresh Dendrobe Juice Consumption on Senile Habitual Constipation for Older People: A Four-Week Randomized Controlled Trial

Background: In Chinese medicine, it is believed that fresh dendrobe juice, which is mild in nature, can relieve the symptom of constipation. Methods: A pilot research design was conducted. Fifty-six older people with senile habitual constipation were recruited and randomly enrolled into control or experimental groups, each with 28 people. The control group was offered water routinely in a day. In addition, the experimental group received 125 mL of dendrobe juice twice a day for four weeks. Quality of life for the old people was evaluated by the Patient Assessment of Constipation Quality of Life (PAC-QOL) Score and the symptom of constipation was assessed by Wexner score. Results: The four-week intervention brought significant performance improvement in all the measured parameters in the experimental group in comparison with the control group. These included significantly more frequency and shorter durations of defecation (p < 0.01, respectively), improved quality of life based on constipation score (PAC-QOL) (experimental group: 50.41 ± 3.46 vs. control group: 70.25 ± 2.35; p < 0.05), and improved score on the Wexner constipation scale (experimental group: 6.56 ± 0.89 vs. control group: 15.50 ± 0.64; p < 0.05). Conclusions: Fresh dendrobe beverage therapy is effective in improving stool frequency, reducing duration of defecation, and enhancing quality of life

Coordinated State Analysis and Differential Regulation of Territorial Spatial Functions in Underdeveloped Regions: A Case Study of Gansu Province, China

The coordinated development of territorial spatial functions is the main manifestation of the sustainable utilization of territorial resources. Identifying spatial functions and their coordinated relationship has become an important guarantee for regional coordinated development, and is of great significance to the construction of a sustainable land development and protection pattern. This study constructs a classification and function scoring system for Production-Living-Ecological Spaces (PLES) by using land-use data from Gansu Province in 2000 and 2020, and revises the spatial functions of the land to supplement the deficiencies of existing related researches by combining socio-economic data (GDP, population density) and ecosystem service value data. In the aforementioned works, the Mechanical Equilibrium Model in physics is referenced to explore the coordinated state among territorial spatial functions. The results show that the high-value areas of production function are mainly distributed in the eastern and central areas of the Loess Plateau, with a strip-like distribution in the Hexi Corridor. The high-value areas of living function are consistent with that in the center of the city. The distribution of high-value areas of ecological function is in line with the topographic distribution pattern of mountainous areas in Gansu Province. The distribution pattern of the coordinated state of land space function in Gansu Province is relatively stable from 2000 to 2020. Simultaneously, the functional dominant area and the promotion area are identified according to the deviation of the coordinated degree of production-living-ecological function. After clarifying the functional characteristics of each county, the author proposes a differentiated regulation strategy of territorial spatial function

Largazole Inhibits Ocular Angiogenesis by Modulating the Expression of VEGFR2 and p21

Ocular angiogenic diseases, characterized by abnormal blood vessel formation in the eye, are the leading cause of blindness. Although Anti-VEGF therapy is the first-line treatment in the market, a substantial number of patients are refractory to it or may develop resistance over time. As uncontrolled proliferation of vascular endothelial cells is one of the characteristic features of pathological neovascularization, we aimed to investigate the role of the class I histone deacetylase (HDAC) inhibitor Largazole, a cyclodepsipeptide from a marine cyanobacterium, in ocular angiogenesis. Our study showed that Largazole strongly inhibits retinal vascular endothelial cell viability, proliferation, and the ability to form tube-like structures. Largazole strongly inhibits the vessel outgrowth from choroidal explants in choroid sprouting assay while it does not affect the quiescent choroidal vasculature. Largazole also inhibits vessel outgrowth from metatarsal bones in metatarsal sprouting assay without affecting pericytes coverage. We further demonstrated a cooperative effect between Largazole and an approved anti-VEGF drug, Alflibercept. Mechanistically, Largazole strongly inhibits the expression of VEGFR2 and leads to an increased expression of cell cycle inhibitor, p21. Taken together, our study provides compelling evidence on the anti-angiogenic role of Largazole that exerts its function through mediating different signaling pathways