Screening, characterization, and optimization of the fermentation conditions of a novel cellulase-producing microorganism from soil of Qinghai-Tibet Plateau.

Cellulases play an important role in the bioconversion of lignocellulose. Microorganisms found in extreme environments are a potentially rich source of cellulases with unique properties. Due to the uniqueness of the environment, the abundant microbial resources in the Qinghai-Tibet Plateau (QTP) are worth being explored. The aim of this study was to isolate and characterize an acidic, mesophilic cellulase-producing microorganism from QTP. Moreover, the fermentation conditions for cellulase production were optimized for future application of cellulase in the development of lignocellulose biomass. A novel cellulase-producing strain, Penicillium oxalicum XC10, was isolated from the soil of QTP. The cellulase produced by XC10 was a mesophilic cellulase that exhibited good acid resistance and some cold-adaptation properties, with maximum activity at pH 4.0 and 40°C. Cellulase activity was significantly enhanced by Na+ (p < 0.05) and inhibited by Mg2+, Ca2+, Cu2+, and Fe3+ (p < 0.05). After optimization, maximum cellulase activity (8.56 U/mL) was increased nearly 10-fold. Optimal fermentation conditions included an inoculum size of 3% (v/v) in a mixture of corn straw (40 g/L), peptone (5 g/L), and Mg2+ (4 g/L), at pH 4.0, 33°C, and shaking at 200 rpm. The specific properties of the P. oxalicum XC10 cellulase suggest the enzyme may serve as an excellent candidate for the bioconversion and utilization of lignocellulose biomass generated as agricultural and food-processing wastes.

Characterization and Assessment of Native Lactic Acid Bacteria from Broiler Intestines for Potential Probiotic Properties.

Probiotics are the most promising alternative to antibiotics for improving animal production and controlling pathogenic infections, while strains derived from natural hosts are considered highly desirable due to their good adaptation to the gastrointestinal tract. The aim of this study was to screen Lactobacillus with broad-spectrum antibacterial activity from broilers fed an antibiotic-free diet and evaluate their potential as poultry probiotics. A total of 44 lactic acid bacteria (LAB) strains were isolated from the intestines of healthy broilers, among which 3 strains exhibited outstanding antimicrobial activity and were subsequently identified through 16S rRNA sequencing as Enterococcus faecium L8, Lactiplantibacillus plantarum L10, and Limosilactobacillus reuteri H11. These three isolates demonstrated potent bacteriostatic activity against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella cholerae, with inhibition zones ranging from 15.67 ± 1.53 to 21.33 ± 0.58 mm. The selected LAB strains exhibited high tolerance to acid and bile salts, with L. reuteri H11 displaying the highest survival rate (ranging from 34.68% to 110.28%) after exposure to 0.3% (w/v) bile salts for 6 h or a low pH environment (pH 2, 2.5, and 3) for 3 h. Notably, L. reuteri H11 outperformed other strains in terms of hydrophobicity (84.31%), auto-aggregation (53.12%), and co-aggregation with E. coli ATCC 25922 (36.81%) and S. aureus ATCC 6538 (40.20%). In addition, the three LAB isolates were either fully or moderately susceptible to the tested antibiotics, except for strain L8, which resisted gentamycin and vancomycin. Consequently, these three LAB strains, especially L. reuteri H11, isolated from the intestines of broiler chickens, represent promising probiotic candidates that can be employed as feed additives to enhance production performance and control poultry pathogens.

Netrin-1 protects blood-brain barrier (BBB) integrity after cerebral ischemia-reperfusion by activating the Kruppel-like factor 2 (KLF2)/occludin pathway.

Ischemia/reperfusion (I/R)-induced neural damage and neuroinflammation have been associated with pathological progression during stroke. Netrin-1 is an important member of the family of laminin-related secreted proteins, which plays an important role in governing axon elongation. However, it is unknown whether Netrin-1 possesses a beneficial role in stroke. Here, we employed the middle cerebral artery occlusion (MCAO) model to study the function of Netrin-1 in alleviating brain injuries. Our results demonstrate that Netrin-1 rescued poststroke neurological deficits and inhibited production of the inflammatory cytokines such as interleukin 6 (IL-6) and endothelial chemokine (C-X-C motif) ligand 1 (Cxcl1). Importantly, Netrin-1 protected against MCAO-induced dysfunction of the blood-brain barrier (BBB) in mice and a reduction in the expression of the tight junction (TJ) protein occludin. Additionally, we report that Netrin-1 could ameliorate oxygen-glucose deprivation/reoxygenation (OGD/R)-induced injury and prevent aggravation in endothelial monolayer permeability in bEnd.3 human brain microvascular endothelial cells (HBMVECs). Mechanistically, Netrin-1 ameliorated OGD/R-induced decrease in occludin and Kruppel-like factor 2 (KLF2) in HBMVECs. Notably, silencing of KLF2 abolished the beneficial effects of Netrin-1 in protecting endothelial permeability and occludin expression, suggesting that these effects are mediated by KLF2. In conclusion, our findings suggest that Netrin-1 could constitute a novel therapeutic strategy for ischemic stroke.

The Effects of Zinc Oxide Nanoparticles on Antioxidation, Inflammation, Tight Junction Integrity, and Apoptosis in Heat-Stressed Bovine Intestinal Epithelial Cells In Vitro.

Zinc oxide nanoparticles (nano-ZnO) have diverse applications in numerous biomedical processes. The present study explored the effects of these nanoparticles on antioxidation, inflammation, tight junction integrity, and apoptosis in heat-stressed bovine intestinal epithelial cells (BIECs). Primary BIECs that were isolated and cultured from calves either were subjected to heat stress alone (42°C for 6 h) or were simultaneously heat-stressed and treated with nano-ZnO (0.8 µg/mL). Cell viability, apoptosis, and expression of genes involved in antioxidation (Nrf2, HO-1, SOD1, and GCLM), inflammation-related genes (TLR4, NF-κB, TNF-α, IL-6, IL-8, and IL-10), intestinal barrier genes (Claudin, Occludin, and ZO-1), and apoptosis-related genes (Cyt-c, Caspase-3, and Caspase-9) were assessed to evaluate the effect of nano-ZnO on heat-stressed BIECs. The nanoparticles significantly increased cell viability and decreased the rate of apoptosis of BIECs induced by heat stress. In addition, nano-ZnO promoted the expression of antioxidant-related genes HO-1 and GCLM and anti-inflammatory cytokine gene IL-10, and inhibited the pro-inflammatory cytokine-related genes IL-6 and IL-8. The nanoparticles also enhanced expression of the Claudin and ZO-1 genes, and decreased expression of the apoptosis-related genes Cyt-c and Caspase-3. These results reveal that nano-ZnO improve the antioxidant and immune capacity of BIECs and mitigate apoptosis of intestinal epithelial cells induced by heat stress. Thus, nano-ZnO have potential for detrimental the adverse effects of heat stress in dairy cows.

Idiopathic pulmonary haemosiderosis misdiagnosed as haemolytic anaemia: a case report.

Idiopathic pulmonary haemosiderosis is a rare disease primarily affecting children. The condition is characterized by widespread bleeding from alveolar capillaries, resulting in symptoms such as haemoptysis, shortness of breath and iron deficiency anaemia. However, it is not a specific disease and sometimes can manifest solely as anaemia, which may be easily overlooked and misdiagnosed. The purpose of this case report was to describe a 1-year-old boy who exhibited haemolytic anaemia as the only symptom of idiopathic pulmonary haemosiderosis, with the intention of offering clinical insights into the precise diagnosis and subsequent management of this rare and easily misdiagnosed disease. Clinicians should keep idiopathic pulmonary haemosiderosis in mind when evaluating children with haemolytic anaemia and promptly initiate testing and treatment to prevent misdiagnosis and improve outcomes.

Effect of guanidinoacetic acid on the growth performance, myofiber, and adenine nucleotide of meat-type rabbits.

OBJECTIVE: This study aimed to investigate the effect of dietary guanidinoacetic acid (GAA) on the growth performance, slaughter traits, myofiber, and adenine nucleotide of meat-type rabbits. METHODS: Experimental treatments consisted of control (CON) and GAA addition at 0.04% (T1), 0.08% (T2), and 0.12% (T3) of diet. A total of 240 weaned rabbits (meat-type male Chinese black rabbits) were randomly distributed into four groups with six replicates of ten rabbits each. RESULTS: Results showed that the three doses of GAA increased (p<0.05) final body weight, carcass weight, the density and area of quadriceps femoris fiber; and T3 showed significant effects (p<0.05) on weight gain, feed/gain, and dressing percentage, and the traits of longissimus fiber, compared to CON. Dietary GAA increased (p<0.05) the meat color a* and b* in longissimus and quadriceps; and T3 showed the lowest (p<0.05) shear force of longissimus. Furthermore, GAA increased (p<0.05) the contents of adenosine triphosphate and total adenine nucleotide in longissimus and quadriceps. In longissimus adenosine triphosphate, total adenine nucleotide, and adenylate energy charges, T3 treatment was most effective (p<0.05); while T2 and T3 treatment was more effective (p<0.05) than T1 in quadriceps. Additionally, linear or quadratic responses (p<0.05) to the increased doses of GAA were found on body weight gain, meat color, total adenine nucleotide, and adenylate energy charges. CONCLUSION: It is concluded that GAA can be used in the rabbit diet to improve growth and carcass traits, and these are related to the high levels of muscle adenine nucleotide.

Novel pneumonia score based on a machine learning model for predicting mortality in pneumonia patients on admission to the intensive care unit.

BACKGROUND: Scores for predicting the long-term mortality of severe pneumonia are lacking. The purpose of this study is to use machine learning methods to develop new pneumonia scores to predict the 1-year mortality and hospital mortality of pneumonia patients on admission to the intensive care unit (ICU). METHODS: The study population was screened from the MIMIC-IV and eICU databases. The main outcomes evaluated were 1-year mortality and hospital mortality in the MIMIC-IV database and hospital mortality in the eICU database. From the full data set, we separated patients diagnosed with community-acquired pneumonia (CAP) and ventilator-associated pneumonia (VAP) for subgroup analysis. We used common shallow machine learning algorithms, including logistic regression, decision tree, random forest, multilayer perceptron and XGBoost. RESULTS: The full data set of the MIMIC-IV database contained 4697 patients, while that of the eICU database contained 13760 patients. We defined a new pneumonia score, the "Integrated CCI-APS", using a multivariate logistic regression model including six variables: metastatic solid tumor, Charlson Comorbidity Index, readmission, congestive heart failure, age, and Acute Physiology Score III. The area under the curve (AUC) and accuracy of the integrated CCI-APS were assessed in three data sets (full, CAP, and VAP) using both the test set derived from the MIMIC-IV database and the external validation set derived from the eICU database. The AUC value ranges in predicting 1-year and hospital mortality were 0.784-0.797 and 0.691-0.780, respectively, and the corresponding accuracy ranges were 0.723-0.725 and 0.641-0.718, respectively. CONCLUSIONS: The main contribution of this study was a benchmark for using machine learning models to build pneumonia scores. Based on the idea of integrated learning, we propose a new integrated CCI-APS score for severe pneumonia. In the prediction of 1-year mortality and hospital mortality, our new pneumonia score outperformed the existing score.

Monoammonium glycyrrhizinate improves antioxidant capacity of calf intestinal epithelial cells exposed to heat stress in vitro.

Dairy calves are highly susceptible to the negative effects of heat stress, which can cause organ hypoxia after blood redistribution, damage the intestinal barrier, and trigger intestinal oxidative stress. This study aimed to investigate the antioxidant effects of monoammonium glycyrrhizinate (MAG) on calf small intestinal epithelial cells under heat stress in vitro. Small intestinal epithelial cells were isolated from a 1-d-old healthy calf and purified by differential enzymatic detachment. The purified cells were divided into seven groups. The control group was cultured with DMEM/F-12 at 37 °C for 6 h, and the treatment groups were cultured with 0, 0.1, 0.25, 0.5, 1, or 5 µg/mL MAG at 42 °C for 6 h. Heat stress causes oxidative damage to cells. Adding MAG to the medium can significantly improve cell activity and reduce cellular oxidative stress. MAG significantly increased the total antioxidant capacity and superoxide dismutase activity caused by heat stress, and significantly decreased malondialdehyde and nitric oxide levels. The MAG treatment also reduced lactate dehydrogenase release, increased mitochondrial membrane potential, and decreased apoptosis under heat stress. MAG also upregulated the expression of the antioxidant-related genes, Nrf2 and GSTT1, in heat-stressed intestinal epithelial cells and significantly downregulated the expression of the heat shock response-related proteins, MAPK, HSP70, HSP90, and HSP27. From the above results, we conclude that 0.25 µg/mL MAG improves the capability of the antioxidant system in small intestinal epithelial cells to eliminate reactive oxygen species by activating antioxidant pathways, improving the oxidant/antioxidant balance, lowering excessive heat shock responses, and reducing intestinal oxidative stress.

In this study, we investigated the antioxidant effect of monoammonium glycyrrhizinate (MAG) on calf intestinal epithelial cells (CIECs) exposed to heat stress in vitro. Calves are sensitive to heat stress, and high temperatures can stimulate heat stress and produce a large number of reactive oxygen species (ROS) to induce oxidative stress. The intestinal tract plays a very important role in the immune defense system of dairy calves. The large amount of ROS can lead to the death of intestinal epithelial cells and damage to intestinal barrier. In order to investigate the antioxidant function of MAG, different concentrations of MAG were added to the culture medium of CIECs and the cells were subsequently exposed to heat stress. The results showed that MAG could effectively relieve oxidative stress and reduce the apoptosis of CIECs exposed to heat stress.

Worldwide Trends in Registering Real-World Studies at ClinicalTrials.gov: A Cross-Sectional Analysis.

Objective: The purpose of this study was to characterize real-world studies (RWSs) registered at ClinicalTrials.gov to help investigators better conduct relevant research in clinical practice. Methods: A retrospective analysis of 944 studies was performed on February 28, 2023. Results: A total of 944 studies were included. The included studies involved a total of 48 countries. China was the leading country in terms of the total number of registered studies (37.9%, 358), followed by the United States (19.7%, 186). Regarding intervention type, 42.4% (400) of the studies involved drugs, and only 9.1% (86) of the studies involved devices. Only 8.5% (80) of the studies mentioned both the detailed study design type and data source in the "Brief Summary". A total of 49.4% (466) of studies had a sample size of 500 participants and above. Overall, 63% (595) of the studies were single-center studies. A total of 213 conditions were covered in the included studies. One-third of the studies (32.7%, 309) involved neoplasms (or tumors). China and the United States were very different regarding the study of different conditions. Conclusion: Although the pandemic has provided new opportunities for RWSs, the rigor of scientific research still needs to be emphasized. Special attention needs to be given to the correct and comprehensive description of the study design in the Brief Summary of registered studies, thereby promoting communication and understanding. In addition, deficiencies in ClinicalTrials.gov registration data remain prominent.

Liver fibrosis is a common pathological change in the liver of dairy cows with fatty liver.

Fatty liver (i.e., hepatic lipidosis) is a prevalent metabolic disorder in dairy cows during the transition period, characterized by excess hepatic accumulation of triglyceride (TG), tissue dysfunction, and cell death. Detailed pathological changes, particularly hepatic fibrosis, during fatty liver remain to be determined. Liver fibrosis occurs as a consequence of liver damage, resulting from the excessive accumulation of extracellular matrix, which distorts the architecture of the normal liver, compromising its normal synthetic and metabolic functions. Thus, we aimed to investigate liver fibrosis status and its potential causal factors including oxidative stress, hepatocyte apoptosis, and production of inflammatory cytokines in the liver of cows with fatty liver. Forty-five dairy cows (parity, 3-5) were selected, and liver biopsy and blood were collected on the second week postpartum (days in milk, 10-14 d). On the basis of the degree of lipid accumulation in liver, selected cows were categorized into normal (n = 25; TG <1% wet wt), mild fatty liver (n = 15; 1% ≤ TG <5% wet wt), and moderate fatty liver (n = 5; 5% ≤ TG <10% wet wt). Compared with normal cows, blood concentrations of nonesterified fatty acids and ß-hydroxybutyrate, along with alanine aminotransferase and aspartate aminotransferase activities, were greater in the cows with fatty liver (mild and moderate). Hepatic extracellular matrix deposition, as indicated by Picrosirius red staining, was greater in cows with fatty liver than those with normal ones. In addition, we observed an increased proportion of collagen type I fiber in extracellular matrix with increased lipid accumulation in the liver. Compared with normal cows, the area of α-smooth muscle actin (α-SMA)-positive staining along with the mRNA abundance of collagen type I α 1 (COL1A1), ACTA2 (gene encoding α-SMA), and transforming growth factor-ß (TGFB) were greater in cows with fatty liver. Compared with normal cows, hepatic contents of malondialdehyde, glutathione disulfide, and 8-isoprostane were greater, whereas total antioxidant capacity, the hepatic content of glutathione, and activities of antioxidant indicators, including superoxide dismutase, glutathione peroxidase, and catalase, were lower in cows with fatty liver. The number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells and abundance of apoptosis-related molecules BAX, CASP3, CASP8, and CASP9 were greater in cows with fatty liver. However, mRNA abundance of the anti-apoptotic gene BCL2 did not differ. The mRNA abundance of pro-inflammatory cytokines including tumor necrosis factor-α (TNFA), interleukin-1ß (IL1B), and interleukin-6 (IL6) was greater in the liver of cows with fatty liver. Overall, the present study indicated that fibrosis is a common pathological response to liver damage and is associated with oxidative stress, hepatocyte death, and inflammation.

Effects of dietary Glycyrrhiza polysaccharide on growth, serum biochemistry, immunity, and egg laying in quail.

This study was conducted to evaluate the effects of dietary supplementation with Glycyrrhiza polysaccharide (GCP) on growth performance, serum biochemistry, immunity, and egg laying in female quail. 300 1-day-old female quail were sorted into four dietary treatments with five replicate cages of 15 birds each. The basic diet in the four treatment groups was supplemented with 0, 500, 1000 and 1500 mg/kg GCP, and the experiment continued for 80 days. Results showed that dietary supplementation with GCP significantly (p < 0.05) increased average daily gain in a dose-dependent fashion, and decreased (p < 0.05) the feed-to-gain ratio and mortality. The relative weights of the thymus and bursa of Fabricius increased (p < 0.05) linearly with increasing dose of GCP from 0 to 1500 mg/kg on day 20. GCP birds showed higher serum levels of protein, glucose, immunoglobulin A and immunoglobulin M, but lower serum triglycerides (p < 0.05) on day 50. GCP increased (p < 0.05) average laying rate and average egg weight linearly from days 60 to 80, whereas feed to egg ratio was decreased (p < 0.05). Taken together, these results revealed that GCP could improve growth performance, serum biochemistry, immunity, and egg laying in female quail. Therefore, GCP may be a potential replacement for antibiotic growth promoters in poultry.

L-arginine reduces injury from heat stress to bovine intestinal epithelial cells by improving antioxidant and inflammatory response.

Heat stress (HS) has a negative impact on the health and performance of dairy cows, resulting in economic losses. Damage to the intestinal epithelium is the main cause of the adverse effects of heat stress on bovine health. This study investigated the repair capability of L-arginine (L-Arg) in reducing the adverse effects of HS on bovine intestinal epithelial cells (BIECs). BIECs were treated as follows: (1) control cells were cultured at 37 °C continuously and received no L-Arg; (2) cells in HS group were grown at 42 °C for 6 h followed by 12 h at 37 °C; and (3) the L-Arg group was cultured at 42 °C for 6 h, then treated with L-Arg at 37 °C for 12 h. HS disrupted redox homeostasis and reduced viability in BIECs, while treatment with L-Arg (6 mmol/L) for 12 h markedly reduced the negative effects of HS. L-Arg protected cells by preventing HS-induced changes in mitochondrial membrane-potential, inflammation, apoptosis-related gene expression and regulation of antioxidant enzymes. The above results indicated that L-Arg reduced the level of damage from HS in BIECs by lowering oxidant stress and inflammation, suggesting that L-Arg could be an effective dietary addition to protect cows from adverse intestinal effects caused by HS.

Effects of rumen-protected methionine supplementation on production performance, apparent digestibility, blood parameters, and ruminal fermentation of lactating Holstein dairy cows.

This study aimed to evaluate the effects of reducing dietary CP and supplementing rumen protected-methionine (RPM) on production performance, blood parameters, digestibility of nutrients or ruminal fermentation in lactating Holstein dairy cows. A total of 96 lactating cows were randomly assigned to 1 of 2 treatments: a diet containing 17.3% CP without RPM (control group; CON; n = 49) or a diet containing 16.4% CP and supplemented with 15.0 g/d of RPM (treatment group; RPM; n = 47). No effect was observed in the RPM group on milk yield, milk composition and digestibility of nutrients. The results of blood parameters showed that cows in the RPM group exhibited lower blood urea nitrogen concentration than in CON group. Rumen microbial crude protein (MCP) was higher in the RPM group compared to the CON group. Ruminal volatile fatty acid (VFA) concentrations were not different between treatments except for butyrate and isovalerate, which were higher in the RPM group than the CON group 2 h after feeding. In conclusion, reducing dietary CP with RPM supplementation did not limit milk yield, milk composition or digestibility of nutrients, but could improve nitrogen utilization, synthesis of MCP and partially increase VFA production 2 h after feeding cows.

Probiotic Properties of Chicken-Derived Highly Adherent Lactic Acid Bacteria and Inhibition of Enteropathogenic Bacteria in Caco-2 Cells.

Lactic acid bacteria (LAB) as probiotic candidates have various beneficial functions, such as regulating gut microbiota, inhibiting intestinal pathogens, and improving gut immunity. The colonization of the intestine is a prerequisite for probiotic function. Therefore, it is necessary to screen the highly adherent LAB. In this study, the cell surface properties, such as hydrophobicity, auto-aggregation, co-aggregation, and adhesion abilities of the six chicken-derived LAB to Caco-2 cells were investigated. All six strains showed different hydrophobicity (21.18-95.27%), auto-aggregation (13.61-30.17%), co-aggregation with Escherichia coli ATCC 25922 (10.23-36.23%), and Salmonella enterica subsp. enterica serovar Typhimurium ATCC 13311 (11.71-39.35%), and adhesion to Caco-2 cells (8.57-26.37%). Pediococcus pentosaceus 2-5 and Lactobacillus reuteri L-3 were identified as the strains with strong adhesion abilities (26.37% and 21.57%, respectively). Moreover, these strains could survive in a gastric acid environment at pH 2, 3, and 4 for 3 h and in a bile salt environment at 0.1%, 0.2%, and 0.3% (w/v) concentration for 6 h. Furthermore, the cell-free supernatant of P. pentosaceus 2-5 and L. reuteri L-3 inhibited the growth of enteropathogenic bacteria and the strains inhibited the adhesion of these pathogens to Caco-2 cells. In this study, these results suggested that P. pentosaceus 2-5 and L. reuteri L-3, isolated from chicken intestines might be good probiotic candidates to be used as feed additives or delivery vehicles of biologically active substances.

Effects of antibacterial peptide-producing Bacillus subtilis, gallic acid, and cellulase on fermentation quality and bacterial community of whole-plant corn silage.

In the current study, we assessed the effects of antibacterial peptide-producing Bacillus subtilis (BS), gallic acid (GA) and cellulase (CL) on the fermentation quality and bacterial community of various varieties of whole-plant corn silage. Three different varieties of whole-plant corn (Yuqing386, Enxiai298, and Nonghe35) were treated with 0.02% BS (fresh material basis), 0.2% GA (fresh material basis) and 0.02% CL (fresh material basis), after which 45 days of anaerobic fermentation were conducted. With the exception of its low dry matter content, the results showed that Yuqing386's crude protein, water-soluble carbohydrate, and lactic acid contents were significantly higher than those of the other two corn varieties. However, its acid detergent fiber and cellulose contents were significantly lower than those of the other two corn varieties. Among the three corn variety silages, Yuqing386 had the highest relative abundance of Lactobacillus at the genus level and the biggest relative abundance of Firmicutes at the phylum level. In addition, the three additives markedly enhanced the quantity of dry matter and crude protein as compared to the control group. The application of GA considerably decreased the level of neutral detergent fiber while significantly increasing the content of lactic acid and water-soluble carbohydrates. Even though all additives enhanced the structure of the bacterial community following silage, the GA group experienced the greatest enhancement. On a phylum and genus level, the GA group contains the highest relative abundance of Firmicutes and Lactobacillus, respectively. Overall, of the three corn varieties, Yuqing386 provides the best silage qualities. GA has the biggest impact among the additions employed in this experiment to enhance the nutritional preservation and fermentation quality of whole-plant corn silage.

Generation of a human induced pluripotent stem cell line (LZUSHi002-A) from a MADD patient with ETFDH mutation.

Multiple acyl-coenzyme A dehydrogenase deficiency (MADD) is an inborn metabolic disorder that affects fatty acid oxidation and the catabolism of branched-chain amino acids, vitamins B and energy metabolism. In this study, the induced pluripotent stem cell (iPSC) line LZUSHi002-A from PBMCs of a 10-year-old male patient with ETFDH mutations using the episomal plasmids was established, which is an ideal in vitro model to understand the exact pathogenesis of MADD.

Protective effects of monoammonium glycyrrhizinate on fatty deposit degeneration induced in primary calf hepatocytes by sodium oleate administration in vitro.

In this study we investigated the effect of MAG on fatty deposit-induced degeneration of primary calf hepatocytes induced by sodium oleate. Primary hepatocytes were isolated from dairy calves and cultured before allocation to the following treatment groups: control (untreated), model (starved for 12 h before treatment with 0.25 mM sodium oleate to induce steatosis-like changes, and the MAG group pretreated with MAG (0.1, 0.25, 0.5, and 1.5 mM) for 12 h before sodium oleate treatment (0.25 mM) for 12 h). To evaluate the effect of MAG on fat-induced degradation of primary hepatocytes, we evaluated lipid deposition, cell viability, apoptosis rate, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity in the culture supernatant, and expression of oxidant and antioxidant enzymes (LDH, MDA, GSH, and CAT), as well as the expression of lipid metabolism-related genes (PPARα, SREBP-1c, ChREBP, CPT1, CPT2, and MTP) and apoptosis-related genes (Cyt-c, Caspase 9, Caspase 8, Caspase 3, Bax, and Bcl-2). MAG significantly reduced lipid accumulation in hepatocytes induced by sodium oleate (P < 0.05), increased cell viability, decreased the apoptosis rate (P < 0.05), and significantly decreased ALT and AST activity in the culture supernatant (P < 0.05). MAG significantly decreased MDA levels in cells and LDH levels in the culture supernatant, while GSH and CAT levels were increased (P < 0.05). MAG significantly increased the expression of the lipid transport- and metabolism-related genes MTP, PPARα, CPT1 and CPT2, and decreased ChREBP expression (P < 0.05). At concentrations higher than 0.25 mM, MAG significantly decreased SREBP-1c expression (P < 0.05). MAG significantly decreased the expression of the apoptosis-related genes Cyt-c, Caspase 9, Caspase 8, Caspase3 and Bax, while Bcl-2 expression was increased (P < 0.05). These findings demonstrate that MAG improves the antioxidant capacity of hepatocytes and effectively reduces lipid deposition by inhibiting the expression of lipid metabolism- and apoptosis-related genes.

Vancomycin, Daptomycin, Antistaphylococcal ß-Lactam, and Trimethoprim-Sulfamethoxazole Monotherapy and Combination Therapy in the Management of Methicillin-Resistant Staphylococcus aureus: A Network Meta-Analysis.

Objective: The aim was to evaluate the efficacy and safety of vancomycin or daptomycin (VAN/DAP), antistaphylococcal ß-lactam (ASBL), trimethoprim-sulfamethoxazole (TMP-SMX), and combination therapy of VAN/DAP + ASBL in the management of methicillin-resistant Staphylococcus aureus (MRSA). Methods: Databases including PubMed, Cochrane Library, Embase database, and google scholar were searched on 1 September 2021. The randomized control trials (RCTs) and comparable clinical studies of VAN/DAP, VAN/DAP + ASBL, ASBL, and TMP-SMX in the management of MRSA were identified. A network meta-analysis was conducted with STATA 14.0. Results: Seven RCTs and two matched cohorts with 1,048 patients were included in the analysis. The pooled results showed that VAN/DAP + ASBL had a significantly lower rate of persistent bacteremia >3 days than VAN/DAP alone [OR:0.46, 95%CI (0.26, 0.81), p < 0.001]. No obvious differences were observed in the outcomes of all-cause mortality, relapsed bacteremia, microbiological treatment failure, embolic or metastatic infection, and total adverse events. However, the ranking results showed that VAN/DAP + ASBL had slightly better efficacy (all-cause mortality, persistent bacteremia >3 days, duration of bacteremia, microbiological treatment failure, and relapsed bacteremia) but slightly higher adverse events than VAN/DAP alone. No obvious differences in the comparisons of VAN/DAP vs. ASBL, and VAN/DAP vs TMP-SMX in the analyzed outcomes. The ranking results revealed that ASBL and TMP-SMX did not have better efficacy or lower adverse events compared with the treatment of VAN/DAP. Conclusion: The efficacy of VAN/DAP + ASBL was slightly but not significantly better than VAN/DAP alone in the management of MRSA.

COVID-19 Clinical Trials Registered Worldwide for Drug Intervention: An Overview and Characteristic Analysis.

OBJECTIVE: This study aims to comprehensively evaluate the characteristics of clinical drug trials to facilitate the collection of evidence for COVID-19 drug treatments. METHODS: A retrospective analysis of 910 trials retrieved on August 7, 2020. RESULTS: A total of 910 registered clinical trials with at least one drug intervention were evaluated. The number of registrations (32.4%, 295) from the United States accounted for nearly one-third of the total and far exceeded that of other countries individually. Furthermore, the peak number of trials were registered in April (34.3%, 312). Over half of the trials (51.2%, 466) are in the recruitment phase, and only 4.2% (38) of the trials have been completed. The median (interquartile range) estimated enrollment is 127 (59, 365). In 39% (355) of trials, the estimated enrollment is less than 100 participants. A total of 94.5% (790) of the trials use randomization in the allocation, 82.7% (753) use a parallel intervention mode, and 52.2% (475) use masking. A total of 287 drug names have been standardized and mapped. "Hydroxychloroquine" is the leading drug among the registered trials (7.47%, 68). Among the main countries contributing to investigations on "hydroxychloroquine", the United States ranks first with 36.76% (25) of the trials. CONCLUSION: The designs of COVID-19 clinical drug trials have greatly improved in terms of the implementation of randomization and, particularly, blinding methods. In terms of drug reuse, the number of drug types has greatly increased, and hundreds of drugs have been used for efficacy screening. The emergence of large-sample registration trials is expected to address the uncertainty regarding the current clinical efficacy of some drugs.

Clinical features and genetic characteristics of hereditary diffuse leukoencephalopathy with spheroids due to CSF1R mutation: a case report and literature review.

BACKGROUND: Hereditary diffuse leukoencephalopathy with spheroid (HDLS) is an autosomal dominant white matter disease characterized by adult-onset cognitive impairment, behavioral or emotional changes, paresis, Parkinsonism, and seizures. Mutations in the gene encoding colony-stimulating factor 1 receptor (CSF1R) have been identified as the cause of HDLS. METHODS: Detail medical history, clinical features and brain imaging of a patient with adult-onset leukoencephalopathy, cognitive impairment and motor dysfunction was reviewed and next generation sequencing was performed. An extensive literature research was then performed to identify all patients with HDLS previously reported. The clinical characteristics, brain imaging and genetic features of patients with HDLS were reviewed. RESULTS: A novel CSF1R mutation, c.1952G>A p.G651E was identified in the patient. Extensive review showed that HDLS typically presents with broad phenotypic variability. The most common symptoms of HDLS were cognitive impairment, followed by psychiatric symptoms, Parkinsonism, gait disorder, and dysphagia. The most common brain imaging findings of HDLS were bilateral white matter lesion, mostly around the ventricles, frontal lobe, and parietal lobe. Calcifications in white matter on CT, cerebral atrophy and thinning of corpus callosum were also common features. Although HDLS demonstrates an autosomal dominant pattern, sporadic cases are not uncommon. CONCLUSIONS: Early recognition of clinical and neuroradiographical characteristics of HDLS is key for the correct diagnosis of the disease.