Coronary artery calcification: concepts and clinical applications.

Vascular calcification is an important hallmark of atherosclerosis. Coronary artery calcification (CAC) implies the presence of coronary artery disease (CAD), irrespective of risk factors or symptoms, is concomitant with the development of advanced atherosclerosis. Coronary thrombosis is the most common clinical end event leading to acute coronary syndrome (ACS). The least common type of pathology associated with thrombosis is the calcified nodule (CN). It usually occurs in elderly patients with severely calcified and tortuous arteries. The prevalence of calcified nodules in patients with ACS may be underestimated due to the lack of easily recognisable diagnostic methods. In this review, the authors will focus on the classification, clinical significance, pathogenesis, and diagnostic evaluation and treatment of CAC to further explore the clinical significance of CN.

Advances in the study of gastric organoids as disease models.

Gastric organoids are models created in the laboratory using stem cells and sophisticated three-dimensional cell culture techniques. These models have shown great promise in providing valuable insights into gastric physiology and advanced disease research. This review comprehensively summarizes and analyzes the research advances in culture methods and techniques for adult stem cells and induced pluripotent stem cell-derived organoids, and patient-derived organoids. The potential value of gastric organoids in studying the pathogenesis of stomach-related diseases and facilitating drug screening is initially discussed. The construction of gastric organoids involves several key steps, including cell extraction and culture, three-dimensional structure formation, and functional expression. Simulating the structure and function of the human stomach by disease modeling with gastric organoids provides a platform to study the mechanism of gastric cancer induction by Helicobacter pylori. In addition, in drug screening and development, gastric organoids can be used as a key tool to evaluate drug efficacy and toxicity in preclinical trials. They can also be used for precision medicine according to the specific conditions of patients with gastric cancer, to assess drug resistance, and to predict the possibility of adverse reactions. However, despite the impressive progress in the field of gastric organoids, there are still many unknowns that need to be addressed, especially in the field of regenerative medicine. Meanwhile, the reproducibility and consistency of organoid cultures are major challenges that must be overcome. These challenges have had a significant impact on the development of gastric organoids. Nonetheless, as technology continues to advance, we can foresee more comprehensive research in the construction of gastric organoids. Such research will provide better solutions for the treatment of stomach-related diseases and personalized medicine.

Characterization of AST-001 non-clinical pharmacokinetics: A novel selective AKR1C3-activated prodrug in mice, rats, and cynomolgus monkeys.

AST-001 is a chemically synthesized inactive nitrogen mustard prodrug that is selectively cleaved to a cytotoxic aziridine (AST-2660) via aldo-keto reductase family 1 member C3 (AKR1C3). The purpose of this study was to investigate the pharmacokinetics and tissue distribution of the prodrug, AST-001, and its active metabolite, AST-2660, in mice, rats, and monkeys. After single and once daily intravenous bolus doses of 1.5, 4.5, and 13.5 mg/kg AST-001 to Sprague-Dawley rats and once daily 1 h intravenous infusions of 0.5, 1.5, and 4.5 mg/kg AST-001 to cynomolgus monkeys, AST-001 exhibited dose-dependent pharmacokinetics and reached peak plasma levels at the end of the infusion. No significant accumulation and gender differences were observed after 7 days of repeated dosing. In rats, the half-life of AST-001 was dose independent and ranged from 4.89 to 5.75 h. In cynomolgus monkeys, the half-life of AST-001 was from 1.66 to 5.56 h and increased with dose. In tissue distribution studies conducted in Sprague-Dawley rats and in liver cancer PDX models in female athymic nude mice implanted with LI6643 or LI6280 HepG2-GFP tumor fragments, AST-001 was extensively distributed to selected tissues. Following a single intravenous dose, AST-001 was not excreted primarily as the prodrug, AST-001 or the metabolite AST-2660 in the urine, feces, and bile. A comprehensive analysis of the preclinical data and inter-species allometric scaling were used to estimate the pharmacokinetic parameters of AST-001 in humans and led to the recommendation of a starting dose of 5 mg/m2 in the first-in-human dose escalation study.

High-resolution free-breathing hepatobiliary phase MRI of the liver using XD-GRASP.

PURPOSE: To evaluate the performance of high-resolution free-breathing (FB) hepatobiliary phase imaging of the liver using the eXtra-Dimension Golden-angle RAdial Sparse Parallel (XD-GRASP) MRI technique. METHODS: Fifty-eight clinical patients (41 males, mean age = 52.9 ± 12.9) with liver lesions who underwent dynamic contrast-enhanced MRI with a liver-specific contrast agent were prospectively recruited for this study. Both breath-hold volumetric interpolated examination (BH-VIBE) imaging and FB imaging were performed during the hepatobiliary phase. FB images were acquired using a stack-of-stars golden-angle radial sequence and were reconstructed using the XD-GRASP method. Two experienced radiologists blinded to acquisition schemes independently scored the overall image quality, liver edge sharpness, hepatic vessel clarity, conspicuity of lesion, and overall artifact level of each image. The non-parametric paired two-tailed Wilcoxon signed-rank test was used for statistical analysis. RESULTS: Compared to BH-VIBE images, XD-GRASP images received significantly higher scores (P < 0.05) for the liver edge sharpness (4.83 ± 0.45 vs 4.29 ± 0.46), the hepatic vessel clarity (4.64 ± 0.67 vs 4.15 ± 0.56) and the conspicuity of lesion (4.75 ± 0.53 vs 4.31 ± 0.50). There were no significant differences (P > 0.05) between BH-VIBE and XD-GRASP images for the overall image quality (4.61 ± 0.50 vs 4.74 ± 0.47) and the overall artifact level (4.13 ± 0.44 vs 4.05 ± 0.61). CONCLUSION: Compared to conventional BH-VIBE MRI, FB radial acquisition combined with XD-GRASP reconstruction facilitates higher spatial resolution imaging of the liver during the hepatobiliary phase. This enhancement can significantly improve the visualization and evaluation of the liver.

A national survey on current state and development needs of clinical and academic emergency medicine in China.

BACKGROUND: To characterize the current state of emergency medicine (EM) and the requirements for advancing EM clinical practice, education and research in China. METHODS: An anonymous electronic survey was conducted by Chinese Society of Emergency Medicine during September to October 2021. The survey contained 30 questions divided into 2 sections: the current state of EM development and the requirements for EM growth. RESULTS: 722 hospitals were included, of 487 were Level III and 235 were Level II hospitals. We found that after 40 years of development, EM had established a mature disciplinary system and refined sub-specialties including critical care, cardiopulmonary resuscitation, toxicology, disaster and emergency rescue. In Level III hospitals, 70.8% of EDs were standardized training centers for EM residents, but master's degree program, Doctor Degree program and post-doctoral degree program was approved in only 37.8%, 8.4% and 2.9% of EDs respectively and postgraduate curriculum was available in 1/4 of EDs. Only 8% have national or provincial key laboratories. In addition to advance clinical practice, there was also a high demand to improve teaching and research capacities, mainly focusing on literature review, research design and delivery, paper writing, residency training. CONCLUSIONS: EM has built a mature discipline system and refined sub-specialties in China. Teaching and research developed parallel with clinical practice. However, there was still a lack of EM master's and doctoral programs and research capacities need to be improved. More outstanding clinical and academic training should be provided to promote the rapid growth of EM in China.

Experimental study of precursory features of CO2 blasting-induced coal rock fracture based on grayscale and texture analysis.

CO2 blasting has been identified as a potent method for enhancing the permeability of coal seams and improving gas drainage efficiency. This study is focused on elucidating the deformation and fracture mechanisms of coal and rock during CO2 blasting and on identifying the precursor characteristics of these processes. To this end, a CO2 blasting-induced coal rock fracture pressure model and a gas pressure distribution model were developed. The research utilized a self-developed CO2 blasting test platform along with a non-contact full-strain field measurement analysis system. Briquette samples were subjected to CO2 blasting tests under controlled experimental conditions, which included an axial pressure of 1.0 MPa and variable gas pressures of 0.5, 1.0, and 1.5 MPa. This methodology enabled the capture of the principal strain field on the surface of the samples. The Gray Level Co-occurrence Matrix (GLCM) was employed to extract and analyze the grayscale and texture features of the strain cloud maps, facilitating a quantitative assessment of their evolution. The aim was to pinpoint the precursor characteristics associated with coal rock cracking and crack propagation. The results revealed that: (1) During the cracking and subsequent propagation of samples, the strain field's grayscale histogram underwent a transformation from a "broad and low" to a "narrow and high" configuration, with a consistent increase in peak frequency. Specifically, at 3 ms, a primary crack was observed in the sample, evidenced by a grayscale peak frequency of 0.0846. By 9 ms, as the crack propagated, the grayscale peak frequency escalated to 0.1626. (2) The texture feature parameters experienced their initial abrupt change at 3ms. Correlation with the gas pressure distribution model indicated that this was the crack initiation moment in the sample. (3) A secondary abrupt shift in the texture feature parameters occurred at 9ms, in conjunction with experimental phenomena, was identified as the crack propagation phase. Monitoring the grayscale and texture features of the principal strain field on the coal rock surface proved effective in recognizing the precursor characteristics of crack initiation and propagation. This research has the potential to reduce blasting costs in coal mines, optimize blasting effects, and provided theoretical guidance for enhancing gas extraction efficiency from deep and low permeability coal seams.

Efficacy of bortezomib, cyclophosphamide, and dexamethasone for newly diagnosed POEMS syndrome patients.

Background: Due to the rarity of polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome, the best first-line treatment has not been established, although there are several options in guidelines. The preferred treatments vary according to the preference of the physician and anecdote. Objectives: First, to analyze the efficacy of a new treatment mode in POEMS syndrome that uses the four-cycle treatment as the induction regimen, followed by sequential transplantation as the consolidation regimen for transplantation-eligible patients, or received another two-cycle treatment for transplantation-ineligible patients. Second, to compare the efficacy and safety of regimens with a proteasome inhibitor (bortezomib-cyclophosphamide-dexamethasone, BCD) or without a proteasome inhibitor (cyclophosphamide-dexamethasone ± thalidomide, CD ± T). Design: We conducted a retrospective study using real-world data from Capital Medical University, Xuanwu Hospital. Methods: A total of 34 newly diagnosed POEMS syndrome patients met Dispenzieri's diagnostic criteria, and those who completed at least four cycles of treatment from July 2013 to March 2021 were included. Results: The overall vascular endothelial growth factor (VEGF) response rate of this new treatment mode was 100%. The cumulative VEGF complete remission (CRV) rate was 67.9%, and the cumulative complete hematological response (CRH) rate was 55.6%. During the median 49-month follow-up, the 5-year-overall survival (OS) rate was 90.7%, the 3-year-progression-free survival (PFS) rate was 78.4%, and the 5-year-PFS rate was 73.8%. The BCD regimen achieved a 75% CRV rate (median time from diagnosis to CRV = 130 days) and 66.7% CRH rate (median time from diagnosis to CRH = 218 days). In addition, the VEGF response was less than the partial remission (PRV) after four-cycle induction treatment, which, together with a decrease on the Overall Neurological Limitation Scale of less than three points 1 year after consolidation treatment, was an independent poor prognostic factor. Conclusion: Bortezomib was well-tolerated by patients with POEMS syndrome. Compared with CD ± T regimen, BCD as the induction regimen achieved better VEGF response and earlier hematological remission. Autologous stem cell transplantation used as consolidation therapy further improved the neurological and hematological remission rates, resulting in better OS and PFS.

Epidemiology and antifungal susceptibility of fungal infections from 2018 to 2021 in Shandong, eastern China: A report from the SPARSS program.

PURPOSE: We analyzed the pathogenic fungal epidemiology and antifungal susceptibility from 2018 to 2021 in Shandong Province, China, to provide the basis for empiric antifungal therapy. METHODS: Fungal isolates were collected from 54 hospitals in Shandong province from 2018 to 2021 through the Shandong Province Pediatric bacterial & fungal Antimicrobial Resistance Surveillance System (SPARSS), WHONET v5.6 and SPSS software v20.0 were used for statistical analysis. RESULTS: A total of 15,348 strains of fungi were collected, with Candida accounting for 78.25 %, followed by Aspergillus at 15.45 %, and other species at 6.27 %. Candida albicans was the predominant Candida species, but more than half of the Candida isolates were non-albicans species, with C. tropicalis being the most dominant (22.74 %), followed by C. glabrata (17.50 %) and C. parapsilosis (11.02 %). The composition of fungi varied significantly among different age groups. Children had a higher proportion of C. albicans (47.30 %) compared to non-children (32.06 %). The non-wild-type phenotype rate of Candida for Amphotericin B was less than 3 %, while Cryptococcus neoformans was 16.67 %. In addition, less than 6 % of C. albicans and C. parapsilosis were resistant to fluconazole and voriconazole, and 96.30 % of C. glabrata were SDD to fluconazole. We also found that 80.56 % of C. glabrata and 83.70 % of C. krusei were voriconazole WT/susceptibility phenotype. However, the susceptibility rates of C. tropicalis to fluconazole/voriconazole decreased from 70.40 %/46.40 % in 2018 to 62.30 %/35.20 % in 2021. The comprehensive susceptibility rate to fluconazole of C. albicans, C. tropicalis, C. parapsilosis and C. glabrata isolated from the blood has decreased from 69.36 % to 56.62 %. CONCLUSIONS: The study reveals that the composition and antifungal susceptibility of pathogenic fungi in Shandong Province differ from other regions. Moreover, the resistance to azoles is more severe, especially in C. tropicalis. These findings indicate the need for region-specific antifungal treatment strategies to combat fungal infections effectively.

Denoising magnetic resonance spectroscopy (MRS) data using stacked autoencoder for improving signal-to-noise ratio and speed of MRS.

BACKGROUND: While magnetic resonance imaging (MRI) provides high resolution anatomical images with sharp soft tissue contrast, magnetic resonance spectroscopy (MRS) enables non-invasive detection and measurement of biochemicals and metabolites. However, MRS has low signal-to-noise ratio (SNR) when concentrations of metabolites are in the range of millimolar. Standard approach of using a high number of signal averaging (NSA) to achieve sufficient SNR comes at the cost of a long acquisition time. PURPOSE: We propose to use deep-learning approaches to denoise MRS data without increasing NSA. This method has potential to reduce the acquisition time as well as improve SNR and quality of spectra, which could enhance the diagnostic value and broaden the clinical applications of MRS. METHODS: The study was conducted using data collected from the brain spectroscopy phantom and human subjects. We utilized a stack auto-encoder (SAE) network to train deep learning models for denoising low NSA data (NSA = 1, 2, 4, 8, and 16) randomly truncated from high SNR data collected with high NSA (NSA = 192), which were also used to obtain the ground truth. We applied both self-supervised and fully-supervised training approaches and compared their performance of denoising low NSA data based on improvement in SNR. To prevent overfitting, the SAE network was trained in a patch-based manner. We then tested the denoising methods on noise-containing data collected from the phantom and human subjects, including data from brain tumor patients. We evaluated their performance by comparing the SNR levels and mean squared errors (MSEs) calculated for the whole spectra against high SNR "ground truth", as well as the value of chemical shift of N-acetyl-aspartate (NAA) before and after denoising. RESULTS: With the SAE model, the SNR of low NSA data (NSA = 1) obtained from the phantom increased by 28.5% and the MSE decreased by 42.9%. For low NSA data of the human parietal and temporal lobes, the SNR increased by 32.9% and the MSE decreased by 63.1%. In all cases, the chemical shift of NAA in the denoised spectra closely matched with the high SNR spectra without significant distortion to the spectra after denoising. Furthermore, the denoising performance of the SAE model was more effective in denoising spectra with higher noise levels. CONCLUSIONS: The reported SAE denoising method is a model-free approach to enhance the SNR of MRS data collected with low NSA. With the denoising capability, it is possible to acquire MRS data with a few NSA, shortening the scan time while maintaining adequate spectroscopic information for detecting and quantifying the metabolites of interest. This approach has the potential to improve the efficiency and effectiveness of clinical MRS data acquisition by reducing the scan time and increasing the quality of spectroscopic data.

Effectiveness of Different Telerehabilitation Strategies on Pain and Physical Function in Patients With Knee Osteoarthritis: Systematic Review and Meta-Analysis.

BACKGROUND: Knee osteoarthritis (OA) is a chronic, degenerative bone and joint disease. It can lead to major pressure to the quality of life and mental health of patients, and also brings a serious economic burden to society. However, it is difficult for patients with knee OA to access rehabilitation when discharging from the hospital. Internet-based rehabilitation is one of the promising telemedicine strategies for the improvement of knee OA, but the effect of different telerehabilitation strategies on knee OA is not clear. OBJECTIVE: The aim of this systematic review and meta-analysis was to identify telerehabilitation strategies attributing to the improvement of pain and physical function outcomes in patients with knee OA. METHODS: We reviewed and analyzed telerehabilitation strategies from randomized controlled trials (RCTs) comparing telerehabilitation with conventional treatment or usual care. For each strategy, we examined whether RCTs that applied the telerehabilitation strategy resulted in a significant improvement in pain or physical function compared with conventional treatment or usual care. RESULTS: We included 6 RCTs (n=734) incorporating 8 different telerehabilitation strategies. The duration of the interventions ranged from 1 to 48 weeks, and sample sizes ranged from 20 to 350 patients. The results showed that RCTs that provided telerehabilitation were found to be more effective than conventional treatments for improving pain (P=.003; standardized mean difference [SMD] -0.21, 95% CI -0.35 to -0.07), but not physical function (P=.24; SMD -0.09, 95% CI -0.25 to 0.06). Furthermore, this systematic review and meta-analysis indicated that there is no significant correlation between different telerehabilitation strategies and the pain and physical function of patients with knee OA. CONCLUSIONS: This systematic review and meta-analysis showed that telerehabilitation programs could relieve pain but not improve physical function for patients with knee OA. These results indicated that telerehabilitation is beneficial for the implementation of home rehabilitation exercises for patients with knee OA, thereby reducing the economic burden of health. However, there were limitations in terms of the number of search results and the number of studies that were eligible for this review and meta-analysis. Therefore, the results need to be interpreted with caution, and more high-quality studies with large samples are needed to focus on the long-term outcomes of telerehabilitation for patients with knee OA to address this limitation.

Impact of percutaneous coronary intervention on renal function in patients with coronary heart disease.

The relationship between cardiac and renal function is complicated. The impact of percutaneous coronary intervention (PCI) on renal function in patients with coronary artery disease is still unclear. The current study sought to assess renal function change, including the time course of renal function, after elective PCI in patients with improved renal function and to identify renal function predictors of major adverse cardiovascular events. We examined data from 1572 CHD patients who had coronary angiography (CAG) or PCI in this retrospective cohort study. Patients receiving elective PCI (n=1240) and CAG (n=332) between January 2013 and December 2018 were included. Pre-PCI and procedural variables associated with post-PCI eGFR, change in renal function after post-PCI follow-up, and post-PCI eGFR association with major adverse cardiovascular events were investigated. Following the procedure, 88.7 percent of PCI group patients had unchanged or improved renal function. The treatment of PCI was found to independently correlate with IRF following coronary angiography in an analysis of patients undergoing PCI [OR 4.561 (95% CI:2 .556-8.139); p<0.001]. The area under the receiver operating characteristic (ROC) curve is 0.763 (model with the treatment of PCI). Improved renal function (IRF) and stable renal function were both associated with a lower risk of a major cardiovascular event.

Clinical value of quantitative analysis of contrast-enhanced ultrasonography in the differential diagnosis of benign and malignant pelvic tumors.

Background: Cervical cancer, endometrial cancer, and ovarian cancer are among the top 10 most common cancers in women, with ovarian cancer in particular being considered a "silent killer". Therefore, early detection, diagnosis, and treatment constitute important means of care for women's health. This study investigated the clinical value of the quantitative analysis of contrast-enhanced ultrasonography (CEUS) in the differential diagnosis of benign and malignant pelvic tumors. Methods: CEUS was performed on 151 patients with pelvic masses. Subsequently, a qualitative diagnosis was completed using the image enhancement features and tumor parameters. A multiparametric analysis of CEUS images was performed, which included the following parameters: arrival time (AT), time to peak (TTP), peak intensity (PI), and ascent slope (AS). In addition, the qualitative diagnostic efficiency of CEUS was assessed in a multiparametric analysis, and the results were compared with pathological findings. Results: The patients in the malignant group were older (P=0.001) and had larger lesion PI values (P<0.01) than those in the benign group. The PI difference (PId) and the AS difference (ASd) showed statistical differences (P<0.01) between the myometrium and lesion tissues in the same patient. Moreover, the PId and ASd showed the largest receiver operating characteristic (ROC) curve and area under the ROC curve (AUC), with sensitivities of 90.9% and 91.7% and specificities of 86.4% and 72.5%, respectively. Conclusions: The quantitative analysis of CEUS provides a new, simpler, and more accurate method for the differential diagnosis of benign and malignant pelvic masses in clinical practice. The sensitivities and specificities of PId and ASd were higher compared to other parameters from the same patient.

Treatment of multidrug-resistant Klebsiella pneumoniae pneumonia in rats with the Wen Run Fei Ning formula: A preliminary study.

BACKGROUND & OBJECTIVES: To determine the effect of Wen Run Fei Ning formula (WRFNF) intervention in class I integron-mediated carbapenem-resistant Klebsiella pneumoniae. METHODS: A drug-susceptibility test and PCR amplification were used to screen for carbapenem-resistant K. pneumoniae containing class I integrons. Following nasal drip and tail vein injection to infect healthy male rats with carbapenem-resistant K. pneumoniae, three models were created: control (group A); model (group B, tail vein injection); and model-WRFNF treatment group (group C, by tail vein injection). Rats in Group C were gavaged with pre-warmed WRFNF extract. On the third, fifth, and seventh days after the experiment, the rats in groups A and B were gavaged with an equal quantity of saline and killed in batches. RESULTS: Group C showed considerably higher serum IL-6 and TNF- levels on days 3, 5, and 7 compared to group A, as well as a significant increase in peripheral blood leukocyte count and a histopathologic inflammatory cell infiltration of the lungs. As the WRFNF delivery duration was prolonged, group C's histopathologic inflammatory cell infiltration gradually improved in contrast to group B, with the biggest improvement occurring on day 7. Compared to group B, group C's serum IL-6 and TNF- levels were lower. When the trial's duration was increased to 7 days, the levels of IL-6 and TNF- in group C decreased on day 7 compared to on day 5. INTERPRETATION & CONCLUSION: WRFNF decreased inflammatory cell infiltration as well as IL-6 and TNF expression in the lung of the rats infected with carbapenem-resistant K. pneumoniae.

Machine learning model for the prediction of gram-positive and gram-negative bacterial bloodstream infection based on routine laboratory parameters.

BACKGROUND: Bacterial bloodstream infection is responsible for the majority of cases of sepsis and septic shock. Early recognition of the causative pathogen is pivotal for administration of adequate empiric antibiotic therapy and for the survival of the patients. In this study, we developed a feasible machine learning (ML) model to predict gram-positive and gram-negative bacteremia based on routine laboratory parameters. METHODS: Data for 2118 patients with bacteremia were obtained from the Medical Information Mart for Intensive Care dataset. Patients were randomly split into the training set and test set by stratified sampling, and 374 routine laboratory blood test variables were retrieved. Variables with missing values in more than 40% of the patients were excluded. Pearson correlation test was employed to eliminate redundant features. Five ML algorithms were used to build the model based on the selected features. Additionally, 132 patients with bacteremia who were treated at Qilu Hospital of Shandong University were included in an independent test cohort to evaluate the model. RESULTS: After feature selection, 32 variables remained. All the five ML algorithms performed well in terms of discriminating between gram-positive and gram-negative bacteremia, but the performance of convolutional neural network (CNN) and random forest (RF) were better than other three algorithms. Consider of the interpretability of models, RF was chosen for further test (ROC-AUC = 0.768; 95%CI = 0.715-0.798, with a sensitivity of 75.20% and a specificity of 63.79%). To expand the application of the model, a decision tree (DT) was built utilizing the major variables, and it achieved an AUC of 0.679 (95%CI = 0.632-0.723), a sensitivity of 66%, and a specificity of 67.82% in the test cohort. When tested in the Qilu Hospital cohort, the ROC-AUC of the RF and DT models were 0.666 (95%CI = 0.579-0.746) and 0.615 (95%CI = 0.526-0.698), respectively. Finally, a software was developed to make the RF- and DT-based prediction models easily accessible. CONCLUSION: The present ML-based models could effectively discriminate between gram-positive and gram-negative bacteremia based on routine laboratory blood test results. This simple model would be beneficial in terms of guiding timely antibiotic selection and administration in critically ill patients with bacteremia before their pathogen test results are available.

[Seasonal Regulation of Soil Microbial Carbon and Phosphorus Metabolisms in an Apple Orchard: Evidence from the Enzymatic Stoichiometry Method].

Soil microbial carbon (C), nitrogen (N), and phosphorus (P) nutrient requirements and metabolic limitations are closely related to the availability of environmental nutrients. However, it is unclear how manure and chemical fertilization shift nutrient limitations for microbes in terms of the soil enzymatic stoichiometry in an apple orchard. Therefore, based on the long-term experiment located in an apple orchard established in 2008, this study applied the theory and method of soil enzyme stoichiometry to systematically investigate the effects of the combined application of manure and chemical fertilizers on soil C, N, and P turnover-related enzyme activities (ß-1,4-glucosidase, BG; leucine aminopeptidase, NAG; ß-1,4-N-acetylglucosaminidase, LAP; and acid or alkaline phosphatase, PHOS) and their stoichiometric characteristics and analyzed their relationships with environmental factors and microbial carbon use efficiency. The experiment was designed with four treatments, such as, no fertilization input as the control (CK), single application of chemical fertilizer (NPK), combined application of manure and chemical fertilizer (MNPK), and single application of manure (M). The results revealed that:① at different growth stages of fruit trees, the soil microbial biomass C (microC) content of manure fertilizer treatments (MNPK and M) was significantly higher than that of no manure fertilizer treatments (CK and NPK). The content of microbial biomass N (microN) in the NPK, MNPK, and M treatments increased by 89%, 269%, and 213%, respectively, compared with that in CK (P<0.05). ② Compared with those in the fertilization treatments, CK had higher leaf N and P contents (29.8 g·kg-1 and 2.17 g·kg-1) at the germination stage, and the leaf P content at the germination stage alone was significantly negatively correlated with soil available phosphorus (AP) content. ③ Soil enzyme stoichiometry analysis demonstrated that all data points in this study were above the 1:1 line, indicating that microbial communities had a strong phosphorus limitation. The range of vector length and angle was 0.56-0.79 and 59.3°-67.7°, respectively, in the growth period of fruit trees, and the vector angle was >45° in this study, which also reflected the strong phosphorus limitation of microorganisms. ④ RDA and random forest model analysis showed that organic carbon and available nitrogen (AN) were the main physical and chemical factors affecting vector length; AP, AN, and soil water content were the main physical and chemical factors affecting vector angle. Combined with SEM analysis, AN and dissolved organic carbon (DOC) directly affected microC and microN, AP directly affected microP and microN, DOC and AP directly affected vector length, and AP and microN directly affected vector angle. In addition, microbial carbon utilization was positively correlated with vector length and negatively correlated with vector angle. In summary, the combined application of manure and chemical fertilizers regulated microbial carbon and phosphorus metabolism by affecting soil carbon and phosphorus content at different growth stages of fruit trees, thereby affecting microbial carbon utilization. This study provides a scientific basis for manure and chemical fertilizers to improve soil quality and maintain soil health.

FBXL5 promotes lipid accumulation in alcoholic fatty liver disease by promoting the ubiquitination and degradation of TFEB.

BACKGROUND: Alcoholic fatty liver disease (AFLD) is characterized by abnormal lipid droplet accumulation in liver. Epigenetic regulation plays an important role in the pathogenesis of AFLD. Comprehensive bioinformatics analysis revealed that an E3 ubiquitin ligase, F-box and leucine-rich repeats protein 5 (FBXL5), was significantly upregulated in AFLD mice. METHODS: The mouse model of AFLD was established by feeding Lieber-DeCarli liquid diet containing ethanol. An in vitro model of AFLD was established by treating HepG2 cells with ethanol (EtOH). The FBXL5 expression was assessed by quantitative real-time PCR (qRT-PCR) and western blotting assays. The levels of triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lipid accumulation were analyzed by enzyme-linked immunosorbent assay (ELISA) and Nile red staining. RESULTS: The FBXL5 expression was markedly up-regulated in in vivo and in vitro models of AFLD compared with controls. Functionally, FBXL5 knockdown alleviated lipid accumulation in EtOH-treated HepG2 cells. Mechanistically, FBXL5 directly interacted with transcription factor EB (TFEB) and accelerated its ubiquitination-mediated degradation. TFEB knockdown reversed the effect of FBXL5 inhibition on decreasing EtOH-induced lipid accumulation. CONCLUSION: Our data suggest that FBXL5 promotes lipid accumulation in AFLD by promoting the ubiquitination and degradation of TFEB.

Transcription factor c-fos induces the development of premature ovarian insufficiency by regulating MALAT1/miR-22-3p/STAT1 network.

BACKGROUND: The current study attempted to investigate the role of transcription factor c-fos in the development of premature ovarian insufficiency (POI) as well as the underlying mechanism involving the MALAT1/miR-22-3p/STAT1 ceRNA network. METHODS: Bioinformatics analysis was performed to extract POI-related microarray dataset for identifying the target genes. Interaction among c-fos, MALAT1, miR-22-3p, and STAT1 was analyzed. An in vivo POI mouse model was prepared followed by injection of sh-c-fos and sh-STAT1 lentiviruses. Besides, an in vitro POI cell model was constructed to study the regulatory roles of c-fos, MALAT1, miR-22-3p, and STAT1. RESULTS: c-fos, MALAT1, and STAT1 were highly expressed in ovarian tissues from POI mice and CTX-induced KGN cells, while miR-22-3p was poorly expressed. c-fos targeted MALAT1 and promoted MALAT1 transcription. MALAT1 competitively bound to miR-22-3p and miR-22-3p could suppress STAT1 expression. Mechanically, c-fos aggravated ovarian function impairment in POI mice and inhibited KGN cell proliferation through regulation of the MALAT1/miR-22-3p/STAT1 regulatory network. CONCLUSION: Our findings highlighted inducing role of the transcription factor c-fos in POI through modulation of the MALAT1/miR-22-3p/STAT1 ceRNA network.

Improving Blood Culture Quality with a Medical Staff Educational Program: A Prospective Cohort Study.

Purpose: Blood cultures (BCs) are essential laboratory tests for diagnosing blood stream infections. BC diagnostic improvement depends on several factors during the preanalytical phase outside of innovative technologies. In order to evaluate the impact of an educational program on BC quality improvement, a total of 11 hospitals across China were included from June 1st 2020 to January 31st 2021. Methods: Each hospital recruited 3 to 4 wards to participate. The project was divided into three different periods, pre-implementation (baseline), implementation (educational activities administered to the medical staff) and post-implementation (experimental group). The educational program was led by hospital microbiologists and included professional presentations, morning meetings, academic salons, seminars, posters and procedural feedback. Results: The total number of valid BC case report forms was 6299, including 2739 sets during the pre-implementation period and 3560 sets during the post-implementation period. Compared with the pre-implementation period, some indicators, such as the proportion of patients who had 2 sets or more, volume of blood cultured, and BC sets per 1000 patient days, were improved in the post-implementation period (61.2% vs 49.8%, 18.56 vs 16.09 sets, and 8.0 vs 9.0mL). While BC positivity and contamination rates did not change following the educational intervention (10.44% vs 11.97%, 1.86% vs 1.94%, respectively), the proportion of coagulase negative staphylococci-positive samples decreased in BSI patients (6.87% vs 4.28%). Conclusion: Therefore, medical staff education can improve BC quality, especially increasing volume of blood cultured as the most important variable to determine BC positivity, which may lead to improved BSI diagnosis.

Changes in the intestinal microbiota of children with hand, foot, and mouth disease under 3 years old.

This study aimed to clarify the characteristics of intestinal microbiota in children with hand, foot, and mouth disease (HFMD) under 3 years old. Fresh feces were collected from 54 children with HFMD and 30 healthy children. All of them were <3 years old. Sequencing of the 16S rDNA amplicons was performed. Between the 2 groups, the richness, diversity, and structure of the intestinal microbiota were analyzed by α-diversity and ß-diversity. Linear discriminant analysis and LEfSe analyses were used to compare different bacterial classifications. The sex and age of the children in the 2 groups were not statistically significant (P = .92 and P = .98, respectively). Compared to healthy children, the Shannon index, Ace index, and Chao index were lower in children with HFMD (P = .027, P = .012, and P = .012, respectively). Based on the weighted or unweighted UniFrac distance analysis, the structure of the intestinal microbiota in HFMD was also significantly changed (P = .002 and P < .001, respectively). Linear discriminant analysis and LEfSe analysis showed that the changes of key bacteria were manifested as a decrease in Prevotella and Clostridium_XIVa (P < .001 and P < .001, respectively), while Escherichia and Bifidobacterium increased (P = .025 and P = .001, respectively). Children with HFMD under 3 years of age have intestinal microbiota disorder and show a decrease in diversity and richness. The decrease in the abundance of Prevotella and Clostridium, which can produce short-chain fatty acids, is also one of the characteristics of the change. These results can offer a theoretical foundation for the pathogenesis and microecological treatment of HFMD in infants.