Revealing the backbone structures of land-based fulvic acids derived from river and soil through n-butylsilane reduction.

While ultra-high-resolution mass spectrometry has enabled the identification of the molecular composition of dissolved organic matter (DOM), elucidating its molecular structure remains a challenging endeavor. Here, two fulvic acids (FAs), one from river and the other from forest soil, were subjected to reduction using an optimized n-butylsilane (n-BS) reduction method. The reduction products were purified through a combination of liquid-liquid extraction and silica gel column chromatography, resulting in the separation into saturates, aromatics, and polar products. The polar products were analyzed by high-resolution mass spectrometry (HRMS), and the saturates and aromatics were analyzed using gas chromatography-mass spectrometry (GC-MS). HRMS results showed that the number of oxygen atoms and double-bond equivalent (DBE) values of FA decreased after reduction. GC-MS results revealed that a total of 270 hydrocarbon monomers were identified from the reduction products of a single sample, with the highest carbon number of cycloalkanes reaching C33. For the first time, steranes and hopanes were detected in the reduction products, potentially serving as evidence for the existence of carboxyl-rich alicyclic molecule (CRAM) precursors. Additionally, a significant number of polycyclic aromatic hydrocarbons were identified, and the potential sources of various compounds were preliminarily inferred based on their isomers. This study extends the knowledge of the possible backbone structure of the DOM and provides a new potential tool for investigating the origin and transformation mechanisms of DOM.

Effect of copper sulphate on Cryptocaryon irritans based on metabolome analysis.

Cryptocaryon irritans is one of the most harmful marine parasites in mariculture. Copper sulphate is often used to kill parasites and the influence of copper sulphate on the tomont stage of C. irritans was explored in this study. The results showed that excystment rate was not significantly affected when tomonts were exposed to 5 mg/L (76.7%) and 10 mg/L (78.9%) of copper sulphate for 3 h. However, excystment rate was significantly inhibited when exposed to 15 mg/L (33.3%) for 3 h and 5 mg/L (28.9%), 10 mg/L (33.3%) and 15 mg/L (33.3%) for 6 h. After treatment with high concentrations of copper sulphate, the interior of the tomonts was fuzzy under the microscope, and the division process could not be observed. Metabolomic results combined with preliminary transcriptome analysis results showed that the tomonts were induced to produce linoleate, riboflavin, inositol and other substances under the stress of Cu2+ , which affected the antioxidant mechanism of the body. Using MDA content determination and antioxidant enzyme activity analysis, copper sulphate was found to cause oxidative damage to tomonts by affecting the generation of metabolites, leading to the death of tomonts.

[Relationship between physical activity and muscle mass loss among people aged 60 and above in 15 provinces of China in 2015].

OBJECTIVE: To analyze the relationship between physical activity and muscle mass loss among Chinese elderly aged 60 years and above in 15 provinces. METHODS: Data was collected from 2015 China Nutritional Transition Cohort Study. Information on demographic characteristics and physical activity was investigated by questionnaire. Dietary intake was estimated from three consecutive 24-h recalls for each individual. Appendicular skeletal muscle mass(ASM) was assessed using the bioelectrical impedance analysis(BIA). Skeletal muscle mass(SMI) loss was diagnosed according Asian working group of sarcopenia(AWGS) 2019 recommendation(male: SMI <7.0 kg/m~2, female: <5.7 kg/m~2). A total of 4565 participants aged 60 years and above from 15 provinces in China with complete information were included in the final analysis. Different domains(occupational, domestic, travel and leisure), light physical activity(<3.0 METs), moderate physical activity(3.0-5.9 METs) and vigorous physical activity(≥6.0 METs) of physical activity among elderly adults were evaluated. Logistic multilevel model was used to analyze the relationship between low muscle mass and different intensity of physical activity duration. RESULTS: Among all participants aged 60 years and above in 15 provinces of China in 2015, women's participation rate in domestic physical activity was 91.7%, which was higher than men's rate of 62.3%. However, men's participation rate in occupational, travel and leisure physical activity were higher than those of women. The participation rate, duration of light physical activity and moderate physical activity were higher in women than in men(light physical activity: female duration 11.0 h/week vs. male duration 3.5 h/week; female participation rate 89.9% vs. male participation rate 62.8%; moderate physical activity: female duration 7.0 h/week vs. male duration 4.7 h/week; female participation rate 90.6% vs. male participation rate 75.2%). The median vigorous physical activity was 0 h/week in both male and female group. The prevalence of muscle mass loss was 13.9% among subjects. The median weekly light physical activity and moderate physical activity in muscle mass loss group were 5.8 h and 4.7 h, which were lower than in non-muscle mass loss group(9.3 h and 6.0 h). The OR of muscle mass lass was 0.71(95%CI 0.42-0.98, P<0.05) for the male with 3.5-6.9 hours of moderate physical activity per week, compared to moderate physical activity less than 3.5 h/week. And OR of the female was 0.67(95%CI 0.47-0.95, P<0.05). CONCLUSION: The vast majority of physical activity in China among the elderly is domestic and light physical activity. Light physical activity and moderate physical activity duration increments in female and moderate physical activity duration increments in male were associated with decreased risk of muscle mass loss.

Trajectories of energy intake distribution and subsequent risk of hyperglycemia among Chinese adults: findings from the China Health and Nutrition Survey (1997-2018).

AIMS: Few studies have examined the secular trend of the energy intake distribution, and its effect on future risk of hyperglycemia. This study aims to describe trajectories of energy intake distribution over 12 years and relate them to subsequent risk of hyperglycemia over 9 years of follow-up. METHODS: Our study used ten waves of data from the CHNS survey, a population-based longitudinal survey in China, ongoing since 1989. We examined a cohort of adult participants who were free from diabetes but had at least three waves of dietary data from 1997 to 2009. We assessed energy intake using three consecutive 24 h recalls. We used these data to identify trajectory groups of energy intake distribution by multi-trajectory model based on energy intake proportions of breakfast, lunch, and dinner. We followed up participants for hyperglycemia, diabetes, and impaired fasting glucose for 9 years from 2009 to 2018. Outcomes were ascertained with fasting glucose, serum HbA1c, and self-report of diabetes and/or glucose-lowering medication. We estimated relative risk (RR) for hyperglycemia, diabetes, and impaired fasting glucose by identified trajectory groups using multilevel mixed-effects modified Poisson regression with robust (sandwich) estimation of variance. Gender difference was additionally examined. RESULTS: A total of 4417 participants were included. Four trajectory groups were identified, characterized and labeled by "Energy evenly distributed with steady trend group" (Group 1), "Dinner and lunch energy dominant with relatively steady trend group" (Group 2), "Dinner energy dominant with increasing trend and breakfast energy with declining trend group" (Group 3), and "breakfast and dinner energy dominant with increasing trend group" (Group 4). During 48,091 person-years, 1053 cases of incident hyperglycemia occurred, 537 cases of incident diabetes occurred, and 516 cases of impaired fasting glucose occurred. Compared with Group 1, Group 3 was associated with higher subsequent risk of incident hyperglycemia in 9 years of follow-up (RR = 1.28, 95% CI = 1.02, 1.61). No association was found for incident diabetes and impaired fasting glucose. Among males, Group 3 was associated with higher risk of incident hyperglycemia in 9 years of follow-up (RR = 1.44, 95% CI = 1.07, 1.94). No relationship was found in females. CONCLUSIONS: Energy intake distribution characterized by over 40% of energy intake from dinner with a rising trend over years was associated with higher long-term risk of hyperglycemia in Chinese adults.

[Multilevel study on the effects of dietary nutrients on body mass index of Chinese children and adolescents aged 7-17].

OBJECTIVE: Analyze the relationship between dietary nutrients and body mass index of children and adolescents aged 7-17 in China. METHODS: The data comes from the "China Health and Nutrition Survey", and 5562 children and adolescents aged 7-17 who participated in at least one round(2000, 2006, 2011 and 2015) of the survey and had complete dietary and physical measurement survey data were selected as the research objects. A three-level(community-individual-observation level) linear random intercept mixed effect model of body mass index was constructed to analyze the influence of dietary nutrient intake of children and adolescents of different genders in urban and rural areas on their body mass index(BMI).24 hours for 3 consecutive days and family weight accounting were used to evaluate the dietary nutrient intake. RESULTS: BMI of urban children and adolescents is higher than that of rural children and adolescents. The BMI of children and adolescents aged 12-17 is higher than that of children and adolescents aged 7-11. BMI of boys was higher than that of girls, but the difference was statistically significant only in 2011 and 2015. After controlling for confounding factors such as individual level(survey year, age, physical activity and family per capita income) and community level(community urbanization index), the three-level model showed that the BMI of rural boys increased with the increase of cholesterol intake(P<0.01). BMI of urban girls increased with the increase of vitamin B_1 intake(P<0.05) and iron intake(P<0.01). BMI of rural girls increased with the increase of vitamin E intake(P<0.001) and sodium intake(P<0.05). CONCLUSION: There are some differences in dietary nutrients that affect the BMI level of 7-17 years old children and adolescents between urban and rural areas.

[Trends of overweight and obesity among children and adolescents aged 7-17 in 16 provinces of China from 2000 to 2018].

OBJECTIVE: To analyze the distribution characteristics and trends of overweight and obesity in children and adolescents aged 7-17 years in 16 provinces of China. METHODS: Data were collected from China health and nutrition survey in 2000, 2006, 2011, 2015 and 2018, 8398 children and adolescents aged 7-17 with complete data were selected as the research objects. Overweight and obesity were classified by body mass index(BMI) reference norm for screening overweight and obesity in Chinese children and adolescents. The distribution and trend of overweight and obesity among children and adolescents aged 7-17 in 16 provinces of China were analyzed by using Cochran-Armitage trend test and two-level random intercept model. RESULTS: The prevalence of overweight and obesity in children and adolescents aged 7-17 in China showed an upward trend from 2000 to 2018(P<0.05). The prevalence of overweight increased from 5.42% in 2000 to 12.92% in 2018, and the prevalence of obesity increased from 2.45% in 2000 to 12.36% in 2018. The result of multilevel model analysis showed that gender and urban and rural areas had significant effects on centralized BMI(P<0.05), the BMI increase value of boys was greater than that of girls(ß=0.2824), the BMI increase value of children and adolescents in rural areas was greater than that in urban areas(ß=0.5902). CONCLUSION: The prevalence of overweight and obesity among children and adolescents aged 7-17 in 16 provinces of China are increasing.

Scalable and Flexible Unsupervised Feature Selection.

Recently, graph-based unsupervised feature selection algorithms (GUFS) have been shown to efficiently handle prevalent high-dimensional unlabeled data. One common drawback associated with existing graph-based approaches is that they tend to be time-consuming and in need of large storage, especially when faced with the increasing size of data. Research has started using anchors to accelerate graph-based learning model for feature selection, while the hard linear constraint between the data matrix and the lower-dimensional representation is usually overstrict in many applications. In this letter, we propose a flexible linearization model with anchor graph and ℓ21 -norm regularization, which can deal with large-scale data sets and improve the performance of the existing anchor-based method. In addition, the anchor-based graph Laplacian is constructed to characterize the manifold embedding structure by means of a parameter-free adaptive neighbor assignment strategy. An efficient iterative algorithm is developed to address the optimization problem, and we also prove the convergence of the algorithm. Experiments on several public data sets demonstrate the effectiveness and efficiency of the method we propose.

Intrathecal Delivery of ssAAV9-DAO Extends Survival in SOD1G93A ALS Mice.

Amyotrophic lateral sclerosis (ALS) is an adult-onset, irreversible neurodegenerative disease that leads to progressive paralysis and inevitable death 3-5 years after diagnosis. The mechanisms underlying this process remain unknown, but new evidence indicates that accumulating levels of D-serine result from the downregulation of D-amino acid oxidase (DAO) and that this is a novel mechanism that leads to motoneuronal death in ALS via N-methyl-D-aspartate receptor-mediated cell toxicity. Here, we explored a new therapeutic approach to ALS by overexpressing DAO in the lumbar region of the mouse spinal cord using a single stranded adeno-associated virus serotype 9 (ssAAV9) vector. A single intrathecal injection of ssAAV9-DAO was made in SOD1G93A mice, a well-established mouse model of ALS. Treatment resulted in moderate expression of exogenous DAO in motorneurons in the lumbar spinal cord, reduced immunoreactivity of D-serine, alleviated motoneuronal loss and glial activation, and extended survival. The potential mechanisms underlying these effects were associated with the down-regulation of NF-κB and the restoration of the phosphorylation of Akt. In conclusion, administering ssAAV9-DAO may be an effective complementary approach to gene therapy to extend lifespans in symptomatic ALS.

Metabolomic changes in Cryptocaryon irritans from Larimichthys crocea after exposure to copper plate.

Cryptocaryon irritans is a highly detrimental parasite in mariculture, causing significant economic losses to the aquaculture industry of Larimichthys crocea. In recent years, copper and copper alloy materials have been used to kill parasites. In this study, the effect of copper plates on the tomont period of C. irritans was explored. The findings indicated that copper plates effectively eradicated tomonts, resulting in a hatching rate of 0. The metabolomic analysis revealed that a total of 2,663 differentially expressed metabolites (1,032 up-regulated and 1,631 down-regulated) were screened in the positive ion mode, and 2,199 differentially expressed metabolites (840 up-regulated and 1,359 down-regulated) were screened in the negative ion mode. L-arginine and L-aspartic acid could be used as potential biomarkers. Copper plate treatment affected 25 metabolic pathways in the tomont, most notably influencing histidine metabolism, retinol metabolism, the biosynthesis of phenylalanine, tyrosine, and tryptophan, as well as arginine and proline metabolism. It was shown that high concentrations of copper ions caused a certain degree of disruption to the metabolome of tomonts in C. irritans, thereby impacting their metabolic processes. Consequently, this disturbance ultimately leads to the rapid demise of tomonts upon exposure to copper plates. The metabolomic changes observed in this study elucidate the lethal impact of copper on C. irritans tomonts, providing valuable reference data for the prevention and control of C. irritans in aquaculture.

Phytoplankton Diversity, Spatial Patterns, and Photosynthetic Characteristics Under Environmental Gradients and Anthropogenic Influence in the Pearl River Estuary.

The Pearl River Estuary (PRE) is one of the world's most urbanized subtropical coastal systems. It presents a typical environmental gradient suitable for studying estuarine phytoplankton communities' dynamics and photosynthetic physiology. In September 2018, the maximum photochemical quantum yield (Fv/Fm) of phytoplankton in different salinity habitats of PRE (oceanic, estuarine, and freshwater zones) was studied, revealing a complex correlation with the environment. Fv/Fm of phytoplankton ranged from 0.16 to 0.45, with taxa in the upper Lingdingyang found to be more stressed. Community composition and structure were analyzed using 18S rRNA, accompanied by a pigment analysis utilized as a supplementary method. Nonmetric multidimensional scaling analysis indicated differences in the phytoplankton spatial distribution along the estuarine gradients. Specificity-occupancy plots identified different specialist taxa for each salinity habitat. Dinophyta and Haptophyta were the predominant taxa in oceanic areas, while Chlorophyta and Cryptophyta dominated freshwater. Bacillariophyta prevailed across all salinity gradients. Canonical correlation analysis and Mantel tests revealed that temperature, salinity, and elevated nutrient levels (i.e., NO3--N, PO43--P, and SiO32--Si) associated with anthropogenic activities significantly influenced the heterogeneity of community structure. The spatial distribution of phytoplankton, along with in situ photosynthetic characteristics, serves as a foundational basis to access estuarine primary productivity, as well as community function and ecosystem health.

Selenoprotein o as a regulator of macrophage metabolism in selenium deficiency-induced lung inflammation.

Selenium (Se) deficiency induces an inflammatory response in the lungs, but the underlying mechanisms are unknown. Selenoprotein O (SelO) is the largest selenoprotein in terms of molecular weight, yet its potential biological functions have yet to be characterized. Our study revealed that Se deficiency leads to an imbalance in the expression of pro-inflammatory "M1" macrophages and anti-inflammatory "M2" macrophages in alveolar macrophages (AMs) and interstitial macrophages (IMs) and contributed to the development of lung inflammation. Through the analysis of differentially expressed selenoproteins, we identified SelO as a potential regulator of the imbalance in pulmonary macrophage polarization caused by Se deficiency. In vitro experiments showed that SelO knockdown enhanced the polarization of M1 macrophages while suppressing that of M2 macrophages. In addition, SelO knockdown reprogrammed macrophage metabolism to glycolysis, disrupting oxidative phosphorylation (OXPHOS). Mechanistically, SelO primarily targets mitochondrial transcription factor A (TFAM), which plays a crucial role in the transcription and replication of mitochondrial DNA (mtDNA) and is essential for mitochondrial biogenesis and energy metabolism. The deficiency of SelO affects TFAM, resulting in its uncontrolled degradation, which compromises mitochondrial function and energy metabolism. In summary, the findings presented here offer significant theoretical insights into the physiological functions of SelO.

Alterations in nasal airflow and air conditioning after endoscopic nasopharyngectomy for recurrent nasopharyngeal carcinoma: a pilot computational fluid dynamics study.

Endoscopic nasopharyngectomy represents a significant intervention for recurrent nasopharyngeal carcinoma (NPC). Various surgical techniques, including transnasal and transoral approaches, are employed. However, the impact of these procedures on nasal airflow dynamics is not well understood. This computational fluid dynamics (CFD) study aimed to investigate alterations in nasal airflow and air conditioning following endoscopic nasopharyngectomy. A 55-year-old male patient with recurrent NPC was selected, whose CT data were utilized for image reconstruction. A preoperative model and two postoperative models, including the transnasal and transoral approach models, were established. The airflow patterns and various CFD parameters were analyzed. In the postoperative models, the high-speed airflow went along the soft palate and into the nasopharyngeal outlet, and there was the low-speed turbulence in the expanded nasopharyngeal cavity. Compared to the preoperative model, the postoperative models exhibited reductions in surface-to-volume ratio, nasal resistance, airflow velocity and proportion of high wall shear stress regions in nasopharynx. The changing trends of nasopharyngeal air temperature and humidity in the preoperative and transoral models were consistent. The heating and humidification efficiency decreased in the transnasal model compared to the transoral model. The endoscopic nasopharyngectomy for recurrent NPC affects the nasal airflow and warming and humidification function. The transoral approach has less influence on aerodynamics of the upper airway compared to the transnasal approach. From a CFD perspective, the endoscopic nasopharyngectomy does not increase the risk of postoperative complications, including the empty nose syndrome and the carotid blowout syndrome.

Physical exercise-induced circAnks1b upregulation promotes protective endoplasmic reticulum stress and suppresses apoptosis via miR-130b-5p/Pak2 signaling in an ischemic stroke model.

AIMS: Physical exercise (PE) can accelerate post-stroke recovery. This study investigated contributions of circRNAs to PE-induced improvements in post-stroke neurological function. METHODS: Rats subjected to transient middle cerebral artery occlusion were left sedentary or provided running-wheel access for 4 weeks during recovery. CircRNAs from peri-infarct cortex were identified by high-throughput sequencing, and interactions with miRNAs by immunoprecipitation, fluorescence in suit hybridization, and dual-luciferase reporter assays. In vivo circRNA knockdown was achieved using shRNA-AAVs and in vitro overexpression by plasmid transfection. Transmission electron microscopy, western blotting, and TUNEL assays were conducted to explore circRNA contributions to endoplasmic reticulum (ER) stress and neuronal apoptosis. CircRNA levels were measured in plasma from stroke patients by qRT-PCR and associations with neurological scores assessed by Pearson's correlation analysis. RESULTS: PE upregulated circAnks1b, reduced infarct volume, and mitigated neurological dysfunction, while circAnks1b knockdown exacerbated neurological dysfunction and increased infarct size despite PE. CircAnks1b sponged miR-130b-5p, thereby disinhibiting Pak2 expression. Conversely, Pak2 downregulation disrupted PE-mediated protective ER stress, leading to reduced IRE1/XBP1 and heightened apoptosis. Plasma circAnks1b was higher in stroke patients receiving PE than sedentary patients and correlated negatively with neurological scores. CONCLUSIONS: CircAnks1b upregulation may be an effective therapeutic strategy for post-stroke recovery.

Bilateral facial palsy caused by HIV infection: A case report and literature review.

OBJECTIVE: Bilateral facial palsy (BFP) has been identified as a possible neurological complication of human immunodeficiency virus (HIV) infection, but only a limited number of cases have been reported in the literature. The purpose of this study was to deepen our understanding of the etiology of BFP. Case report: We report the case of a 46-year-old married bisexual man with BFP associated with HIV infection. The patient underwent serological testing for HIV and was positive. In the absence of any other evidence of underlying systemic disease, facial palsy is thought to be secondary to HIV infection. After antiretroviral therapy, the patient recovered completely from facial palsy within 3 months. Results: HIV infection often involves BFP. The pathophysiology of this clinical presentation is thought to be related to the immune response to the systemic transmission of the virus. Conclusions: Most patients with BFP have underlying systemic causes, particularly autoimmune diseases. The exclusion of HIV infection in patients with BFP is essential for the early diagnosis and management of HIV.

Exploring the mechanism of the PTP1B inhibitors by molecular dynamics and experimental study.

Protein tyrosine phosphatase 1B (PTP1B) has proven to be an attractive target for the treatment of cancer, diabetes and other diseases. Although many PTP1B inhibitors with various scaffolds have been developed, there is still a lack of PTP1B inhibitor with high specificity and acceptable pharmacological properties. Therefore, it is urgent to develop more methods to explore complex action mode of PTP1B and ligands for designing ideal PTP1B modulators. In this work, we developed a potential molecular dynamics (MD) analytic mode to analyze the mechanism of active compounds 6a and 6e against PTP1B from different perspectives, including the stable ability, interactions and binding site of ligand and protein, the binding energy, relative movement between residues and changes in protein internal interactions. The simulated results demonstrated that compound 6a bound more stably to the active pocket of PTP1B than 6e due to its smaller molecular volume (326 Å3), matched electronegativity, and enhanced the positive correlation motion of residues, especially for WPD loop and P loop. Lastly, compound 6a as a competitive inhibitor for PTP1B was verified by enzyme kinetic assay. This work successfully studied the mechanism of compound 6a against PTP1B from various aspects, enriched the analysis of interaction mode between PTP1B and inhibitors. In summary, we hope that this work could provide more theoretical information for designing and developing more novel and ideal PTP1B inhibitors in the future.

Longitudinal Association of Dietary Energy Density with Abdominal Obesity among Chinese Adults from CHNS 1993-2018.

Few studies have explored the longitudinal association between dietary energy density and waist circumference and abdominal obesity in adults in China. This study aimed to analyze the relationship between dietary energy density and waist circumference and abdominal obesity in Chinese residents aged 18-64. Using data from the CHNS from 1993 to 2018, 25,817 adult residents aged 18 to 64 were selected for the study. Three consecutive 24-h dietary recalls and home-weighed seasonings were used to assess food intake. A multilevel model was used to analyze the relationship between dietary energy density and waist circumference and abdominal obesity. The results showed that compared with the lowest dietary energy density group, females had an increased risk of abdominal obesity (OR = 1.16, 95% CI: 1.05, 1.29), and females' waist circumference increased significantly by 0.24 cm (95% CI: 0.39-1.09) in the highest dietary energy density group. No association between dietary energy density and waist circumference and abdominal obesity was observed in males. This study shows that higher dietary energy density is significantly associated with females' waist circumference and abdominal obesity. Further research on high dietary energy density and abdominal obesity will provide scientific basis for the effective control of abdominal obesity.

Longitudinal Association between Urbanicity and Total Dietary Fat Intake in Adults in Urbanizing China from 1991 to 2015: Findings from the CHNS.

Few studies have examined the longitudinal association between urbanicity and dietary fat intake in Chinese adults. A population-based longitudinal observational study was carried out in Chinese adults aged 18-65 from the China Health and Nutrition Survey. Three consecutive 24 h dietary recalls were used to assess dietary fat intake. Multilevel models were used to explore the relationship between urbanicity and dietary fat intake. People in the highest urbanicity quartile had the increments of 7.48 g/d (95% CI:5.42-9.58) and 8.92 g/d (95% CI: 7.03-10.80) in dietary fat intake, 2.86 (95% CI: 2.29-3.44) and 2.69 (95% CI: 2.13-3.25) in proportion of energy from total fat, and odds ratios (ORs) for the risk of excess dietary fat intake of 1.84 (95% CI: 1.65-2.05) and 2.01 (95% CI: 1.78-2.26) for men and women, respectively, compared to the lowest quartile after controlling for potential confounders. These results indicate that urbanicity was an important factor influencing dietary fat intake among Chinese adults. Aggressive nutritional education action coupled with governmental guidelines and programs tailored for the Chinese population are required to promote less dietary fat intake, especially in those adults living in less urbanized areas and whose dietary fat intake is ≥ 30% of their total energy intake per day.

Intrathecal Injection of scAAV9-hIGF1 Prolongs the Survival of ALS Model Mice by Inhibiting the NF-kB Pathway.

Amyotrophic lateral sclerosis (ALS) is a chronic, fatal neurodegenerative disorder characterized by the progressive loss of upper and lower motor neurons. Currently, there is no effective drug for ALS. Recent studies in ALS model mice have shown that insulin-like growth factor-1 (IGF1) may be a promising therapeutic drug. We demonstrate that self-complementary adeno-associated virus serum type 9 encoding the human IGF1 (scAAV9-hIGF1) could significantly postpone the onset and slow down the progression of the disease owning to inhibiting the NF-κB signaling pathway. Furthermore, the results were supported by experiments in which the CRISPR/Cas9 system was used to knock-down IGF1 in ALS mice (mIGF1). Our data indicate that IGF1-mediated suppression of NF-κB activation in microglia is a novel molecular mechanism underlying MN death in ALS. It provides new insight into IGF1 and points toward novel therapeutic targets of IGF1 in ALS.

Intramuscular Delivery of scAAV9-hIGF1 Prolongs Survival in the hSOD1G93A ALS Mouse Model via Upregulation of D-Amino Acid Oxidase.

Self-complementary adeno-associated viral vector 9 (scAAV9) has been confirmed to be an efficient AAV serotype for gene transfer to the central nervous system (CNS). Neurotrophic factors have been considered to be therapeutic targets for amyotrophic lateral sclerosis (ALS). In the present study, we intramuscularly injected scAAV9 encoding human insulin-like growth factor 1 (hIGF1) into an hSOD1G93A ALS mouse model. We observed that scAAV9-hIGF1 significantly reduced the loss of motor neurons of the anterior horn in the lumbar spinal cord and delayed muscle atrophy in ALS mice. Importantly, IGF1 significantly delayed disease onset and prolonged the life span of ALS mice. In addition, scAAV9-hIGF1 protected motor neurons from apoptosis through upregulation of D-amino acid oxidase (DAO), which controls the level of D-serine. Moreover, to further verify these results, we used CRISPR-Cas9 system to target the central nervous system knockdown of IGF1. This experiment supported the continued investigation of neurotrophic factor gene therapies targeting the central nervous system as a potential treatment for ALS.

scAAV9-VEGF prolongs the survival of transgenic ALS mice by promoting activation of M2 microglia and the PI3K/Akt pathway.

Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease that leads to paralysis and death three to five years after diagnosis in most patients. The disease is incurable, and the mechanism of motoneuron degeneration remains unknown, although research has demonstrated that activated microglia are involved in motor neuron death. Here, we used a simple method to deliver AAV9 virus by direct intrathecal injection and found that scAAV9-VEGF-165 improved the motor performance and prolonged the life span of SOD1-G93A mice. Furthermore, scAAV9-VEGF-165 activated the PI3K/Akt survival pathway and increased the level of Bcl-2, which contributed to the protection of motor neurons. Additionally, scAAV9-VEGF-165 attenuated the expression of classically activated (M1) microglial markers and enhanced the expression of alternatively activated (M2) microglial markers. Taken together, the results of our study suggest that simple, direct intrathecal injection of scAAV9-VEGF-165 may have a curative effect for ALS.