Enhancing the therapeutic efficacy of gefitinib on subcutaneously transplanted SKOV3 ovarian cancer tumors in nude mice via ultrasound­stimulated microbubble cavitation.

The present study aimed to explore the effect of ultrasound-stimulated microbubble cavitation (USMC) on drug concentration and therapeutic efficacy of oral gefitinib in treating subcutaneously transplanted SKOV3 ovarian cancer tumors in nude mice. The present study employed the VINNO70 ultrasonic diagnostic and treatment integrated machine for USMC therapy. Firstly, the mechanical index was set at 0.25, and the therapeutic efficacy of USMC treatment was assessed at intervals of 5, 10 and 20 min. Briefly, 72 nude mice were randomized into the following four groups (n=18/group): Control group, USMC5 min group, USMC10 min group and USMC20 min group, and the therapeutic response to USMC treatment was evaluated by comparing pre-and post-intervention effects. Additionally, the combined therapeutic efficacy of USMC and gefitinib was investigated by randomly dividing 96 tumor-bearing mice into the following four groups (n=24/group): Control group, USMC group, gefitinib group and USMC + gefitinib group. Contrast-enhanced ultrasound, hematoxylin and eosin staining, western blotting, immunofluorescence staining, TUNEL staining, ELISA and liquid chromatography-mass spectrometry were performed in the present study. The results showed that USMC combined with gefitinib had the best treatment effect; the tumor inhibition rate was higher than that of gefitinib alone and the overall survival time was prolonged. In addition, the drug concentration in the tumor tissue obtained from the USMC + gefitinib group was revealed to be ~1.4 times higher than that detected in the group treated with gefitinib alone. The experimental results also confirmed that the strongest tumor inhibition rate and longest overall survival time was observed in the USMC + gefitinib group, followed by the gefitinib group and USMC group. STAT3 is an important signaling transducer and transcription factor, which, when phosphorylated, can lead to abnormal cell proliferation and malignant transformation. In addition, the upregulation of phosphorylated (p)-STAT3 is consider a reason for the poor efficacy of gefitinib in treating ovarian cancer. The present study revealed that ultrasound microbubble therapy could overcome this side effect. In conclusion, USMC improved the effects of oral gefitinib on subcutaneously transplanted SKOV3 ovarian cancer tumors in nude mice and increased drug penetration. In addition, USMC overcame the gefitinib-induced side effect of upregulated STAT3 phosphorylation and reduced the expression levels of p-STAT3 in the tumor.

The application of shear wave quantitative ultrasound elastography in chronic kidney disease.

BACKGROUND: Chronic kidney disease (CKD) is a major public health problem, so it is particularly important to quantitatively assess and intervene in the degree of early renal damage in CKD. OBJECTIVE: The objective of the research is to establish reference values for kidney elasticity by using real-time shear wave elastography (RT-SWE) technology to quantify Young's modulus values in the renal cortex of normal adults. The intention is to provide a foundation for evaluating renal function and structural changes in patients with CKD. Furthermore, this research investigates the role of RT-SWE in the early detection of renal fibrosis in CKD, providing insights into its diagnostic value for detecting pathological changes at an early stage. METHODS: Between August 2019 and December 2021, we collected a sample of 100 healthy people (55 men with an average age of 43.5 ± 15.2 years and 45 women with an average age of 41.6 ± 19.8 years) for medical evaluations at our hospital's Department of Ultrasound Medicine. In addition, 97 individuals with CKD1-3 stage were considered. Following the removal of contraindications and relevant confounding variables, we included a final cohort of 80 individuals in the research (45 men and 35 females, with an average age of 39.1 ± 19.2 years). The RENAL mode was selected and a convex array probe S6-1 operating at a frequency of 3.5-5.5 MHz was used in the research, which made use of the French Supersonic AixPlorer ultrasonic diagnostic instrument. Renal RT-SWE elastography was performed after conventional two-dimensional and color Doppler ultrasonography. The study used RT-SWE technology to assess the mean Young's modulus of the cortex in healthy individuals (Emean), with data analysis and comparisons based on age and gender. Furthermore, the Emean values of CKD stage 1-3 patients were determined, and analyses were performed about 24-hour urine protein quantitative (24hUTP), serum creatinine concentration (SCr), and renal biopsy pathology, specifically the degree of interstitial fibrosis. RESULTS: Healthy group: a) The average kPa values of the left kidney (4.2 ± 2.3), right kidney (4.3 + 2.5) kPa, both kidneys' average kPa values (4.3 ± 2.4) kPa, and the average kPa values of the left and right kidneys do not differ statistically (p= 0.986). b) There was no difference in the kPa values of healthy male and female kidneys (4.4 + 2.1 and 4.2 + 2.6, respectively. c) There was no difference in the renal kPa values of healthy adults aged 50 (4.4 ± 2.8) kPa and renal kPa of the 50-year-old population (4.2 + 2.1) kPa (p= 0.041). Case group: a) the group of patients with CKD1-3 stage and the group did not vary in their Emean values (both p< 0.05); b) There is a difference between CKD stages 1, 2, and 3 (p< 0.05), however, there is still no difference in the pyEmean value corrected for patient age between patients in stages 1 and 2 (p> 0.05). CONCLUSION: The study reveals no significant differences in the Emean value of bilateral kidneys in normal people and no differences in the elasticity value of kidneys and gender. However, age-based differences were statistically significant. pyEmean may be useful for comparing CKD stage 1, 2, and 3 patients, and RT-SWE can assess early renal damage.

Enhancement of Tumor Perfusion and Antiangiogenic Therapy in Murine Models of Clear Cell Renal Cell Carcinoma Using Ultrasound-Stimulated Microbubbles.

OBJECTIVE: To explore the effect of ultrasound-stimulated microbubble cavitation (USMC) on enhancing antiangiogenic therapy in clear cell renal cell carcinoma. MATERIALS AND METHODS: We explored the effects of USMC with different mechanical indices (MIs) on tumor perfusion, 36 786-O tumor-bearing nude mice were randomly assigned into four groups: (i) control group, (ii) USMC0.25 group (MI = 0.25), (iii) USMC1.4 group (MI = 1.4) (iv) US1.4 group (MI = 1.4). Tumor perfusion was assessed by contrast-enhanced ultrasound (CEUS) before the USMC treatment and 30 min, 4h and 6h after the USMC treatment, respectively. Then we evaluated vascular normalization(VN) induced by low-MI (0.25) USMC treatment, 12 tumor-bearing nude mice were randomly divided into two groups: (i) control group (ii) USMC0.25 group. USMC treatment was performed, and tumor microvascular imaging and blood perfusion were analyzed by MicroFlow imaging (MFI) and CEUS 30 min after each treatment. In combination therapy, a total of 144 tumor-bearing nude mice were randomly assigned to six groups (n = 24): (i) control group, (ii) USMC1.4 group, (iii) USMC0.25 group, (iv) bevacizumab(BEV) group, (v) USMC1.4 +BEV group, (vi) USMC0.25 +BEV group. BEV was injected on the 6th, 10th, 14th, and 18th d after the tumors were inoculated, while USMC treatment was performed 24 h before and after every BEV administration. We examined the effects of the combination therapy through a series of experiments. RESULTS: Tumor blood perfusion enhanced by USMC with low MI (0.25)could last for more than 6h, inducing tumor VN and promoting drug delivery. Compared with other groups, USMC0.25+BEV combination therapy had the strongest inhibition on tumor growth, led to the longest survival time of the mice. CONCLUSION: The optimized USMC is a promising therapeutic approach that can be combined with antiangiogenic therapy to combat tumor progression.

PFKFB3-driven vascular smooth muscle cell glycolysis promotes vascular calcification via the altered FoxO3 and lactate production.

A link between increased glycolysis and vascular calcification has recently been reported, but it remains unclear how increased glycolysis contributes to vascular calcification. We therefore investigated the role of PFKFB3, a critical enzyme of glycolysis, in vascular calcification. We found that PFKFB3 expression was upregulated in calcified mouse VSMCs and arteries. We showed that expression of miR-26a-5p and miR-26b-5p in calcified mouse arteries was significantly decreased, and a negative correlation between Pfkfb3 mRNA expression and miR-26a-5p or miR-26b-5p was seen in these samples. Overexpression of miR-26a/b-5p significantly inhibited PFKFB3 expression in VSMCs. Intriguingly, pharmacological inhibition of PFKFB3 using PFK15 or knockdown of PFKFB3 ameliorated vascular calcification in vD3 -overloaded mice in vivo or attenuated high phosphate (Pi)-induced VSMC calcification in vitro. Consistently, knockdown of PFKFB3 significantly reduced glycolysis and osteogenic transdifferentiation of VSMCs, whereas overexpression of PFKFB3 in VSMCs induced the opposite effects. RNA-seq analysis and subsequent experiments revealed that silencing of PFKFB3 inhibited FoxO3 expression in VSMCs. Silencing of FoxO3 phenocopied the effects of PFKFB3 depletion on Ocn and Opg expression but not Alpl in VSMCs. Pyruvate or lactate supplementation, the product of glycolysis, reversed the PFKFB3 depletion-mediated effects on ALP activity and OPG protein expression in VSMCs. Our results reveal that blockade of PFKFB3-mediated glycolysis inhibits vascular calcification in vitro and in vivo. Mechanistically, we show that FoxO3 and lactate production are involved in PFKFB3-driven osteogenic transdifferentiation of VSMCs. PFKFB3 may be a promising therapeutic target for the treatment of vascular calcification.

Effects of first-line antidiabetic drugs on the improvement of arterial stiffness: A Bayesian network meta-analysis.

BACKGROUND: Changes in vascular function are closely associated with the development of cardiovascular disease (CVD). Pulse wave velocity (PWV) is a potential indicator of vascular dysfunction; it allows noninvasive assessment of arterial stiffness. Currently, evidence for the effects of different classes of antidiabetic drugs on arterial stiffness remains limited. In this study, a network meta-analysis (NMA) was performed to explore the associations between changes in arterial stiffness and first-line antidiabetic drugs by evaluating PWV in patients with different metabolic abnormalities. METHODS: We systematically searched several electronic databases for randomized controlled trials (RCTs) published from inception until 25 August 2022, without language restrictions. The primary outcome was the change in PWV (ΔPWV) in all included studies; subgroup analysis was performed for patients with abnormal glucose metabolism, including prediabetes and diabetes mellitus. NMA was performed to calculate the mean differences (MDs) with 95% confidence intervals (CIs) as effect sizes to evaluate the ΔPWV. RESULTS: Among the 2257 candidate articles identified in the initial search, 18 RCTs were eventually included in the analysis. In all studies, two classes of new antidiabetic drugs, glucagon-like peptide-1 receptor (GLP-1R) agonists and sSodium-glucose co-transporter 2 (SGLT-2) inhibitors, improved arterial stiffness by decreasing PWV compared with placebo (MD = -1.11, 95% CI: -1.94 to 0.28) and (MD = -0.76, 95% CI: -1.45 to -0.08). A conventional antidiabetic drug, metformin, also showed similar efficacy compared with placebo (MD = -0.73, 95% CI: -1.33 to -0.12). Finally, in subgroup studies of patients with abnormal glucose metabolism diseases, GLP-1R agonists (MD = -1.06, 95% CI: -2.05 to -0.10) significantly decreased PWV compared with placebo. CONCLUSION: Three classes of antidiabetic drugs-GLP-1R agonists, SGLT-2 inhibitors, and metformin-have the potential to improve arterial stiffness. Among the six classes of antidiabetic drugs analyzed, GLP-1R agonists constitute the only class of drugs that improves arterial stiffness in patients with abnormal glucose metabolism diseases.

A novel NSUN5/ENO3 pathway promotes the Warburg effect and cell growth in clear cell renal cell carcinoma by 5-methylcytosine-stabilized ENO3 mRNA.

OBJECTIVES: Clear cell renal cell carcinoma (ccRCC) cells often reprogram their metabolisms. Enolase 3 (ENO3) is closely related to the Warburg effect observed in cells during tumor progression. However, the expression and function of ENO3 in ccRCC cells remain unclear. Therefore, this study investigated the expression and functional significance of ENO3 in the Warburg effect observed in ccRCC cells. METHODS: In this study, B-mode and microflow imaging ultrasound examinations were performed to evaluate patients with ccRCC. The extracellular acidification rate test and glucose uptake and lactate production assays were used to examine the Warburg effect in ccRCC cells. Western blotting, quantitative reverse transcription polymerase chain reaction, and immunochemistry were used to detect the expression of ENO3 and NOP2/Sun RNA methyltransferase 5 (NSUN5). RESULTS: ENO3 upregulation in ccRCC tumor tissues was accompanied by an increase in tumor size. Importantly, ENO3 participated in the Warburg effect observed in ccRCC cells, and high levels of ENO3 indicated a poor prognosis for patients. Loss of ENO3 reduced glucose uptake, lactate production, and extracellular acidification rate as well as inhibited ccRCC cell proliferation. Furthermore, NSUN5 was involved in the ENO3-regulated Warburg effect and ccRCC cell progression. Mechanically, NSUN5 was upregulated in ccRCC tissues, and NSUN5 upregulation mediated 5-methylcytosine modification of messenger RNA (mRNA) in ccRCC cells to promote mRNA stability and ENO3 expression. CONCLUSIONS: Collectively, the destruction of the NSUN5/ENO3 axis prevents ccRCC growth in vivo and in vitro, and targeting this pathway may be an effective strategy against ccRCC progression.

Effects of Organophosphate-Degrading Bacteria on the Plant Biomass, Active Medicinal Components, and Soil Phosphorus Levels of Paris polyphylla var. yunnanensis.

Paris polyphylla var. yunnanensis, a medicinal plant that originated in Yunnan (China), has been over-harvested in the wild population, resulting in its artificial cultivation. Given the negative environmental impacts of the excessive use of phosphorus (P) fertilization, the application of organophosphate-degrading bacteria (OPDB) is a sustainable approach for improving the P use efficiency in Paris polyphylla var. yunnanensis production. The present work aimed to analyze the effects of three organic phosphate-solubilizing bacteria of Bacillus on the yield and quality of P. polyphylla var. yunnanensis and the P concentrations in the soil. All the inoculation treatments distinctly increased the rhizome biomass, steroidal, and total saponin concentrations of the rhizomes and the Olsen-P and organic P in the soil. The highest growth rate of rhizomes biomass, steroidal saponins, available phosphorus, and total phosphorus content was seen in the S7 group, which was inoculated with all three OPDB strains, showing increases of 134.58%, 132.56%, 51.64%, and 17.19%, respectively. The highest total saponin content was found in the group inoculated with B. mycoides and B. wiedmannii, which increased by 33.68%. Moreover, the highest organic P content was seen in the group inoculated with B. wiedmannii and B. proteolyticus, which increased by 96.20%. In addition, the rhizome biomass was significantly positively correlated with the saponin concentration, together with the positive correlation between the Olsen-P and organic P and total P. It is concluded that inoculation with organophosphate-degrading bacteria improved the biomass and medicinal ingredients of the rhizome in P. polyphylla var. yunnanensis, coupled with increased soil P fertility, with a mixture of the three bacteria performing best.

Development and validation of a new predictive model for macrosomia at late-term pregnancy: A prospective study.

Objective: Fetal macrosomia is defined as a birth weight more than 4,000 g and is associated with maternal and fetal complications. This early metabolic disease may influence the entire life of the infant. Currently, macrosomia is predicted by using the estimated fetal weight (EFW). However, the EFW is inaccurate when the gestational week is gradually increasing. To assess precisely the risk of macrosomia, we developed a new predictive model to estimate the risk of macrosomia. Methods: We continuously collected data on 655 subjects who attended regular antenatal visits and delivered at the Second Hospital of Hebei Medical University (Shijiazhuang, China) from November 2020 to September 2021. A total of 17 maternal features and 2 fetal ultrasonographic features were included at late-term pregnancy. The 655 subjects were divided into a model training set and an internal validation set. Then, 450 pregnant women were recruited from Handan Central Hospital (Handan, China) from November 2021 to March 2022 as the external validation set. The least absolute shrinkage and selection operator method was used to select the most appropriate predictive features and optimize them via 10-fold cross-validation. The multivariate logistical regressions were used to build the predictive model. Receiver operating characteristic (ROC) curves, C-indices, and calibration plots were obtained to assess model discrimination and accuracy. The model's clinical utility was evaluated via decision curve analysis (DCA). Results: Four predictors were finally included to develop this new model: prepregnancy obesity (prepregnancy body mass index ≥ 30 kg/m2), hypertriglyceridemia, gestational diabetes mellitus, and fetal abdominal circumference. This model afforded moderate predictive power [area under the ROC curve 0.788 (95% confidence interval [CI] 0.736, 0.840) for the training set, 0.819 (95% CI 0.744,0.894) for the internal validation set, and 0.773 (95% CI 0.713,0.833) for the external validation set]. On DCA, the model evidenced a good fit with, and positive net benefits for, both the internal and external validation sets. Conclusions: We developed a predictive model for macrosomia and performed external validation in other regions to further prove the discrimination and accuracy of this predictive model. This novel model will aid clinicians in easily identifying those at high risk of macrosomia and assist obstetricians to plan accordingly.

Preparation of ultrasound contrast agents: The exploration of the structure-echogenicity relationship of contrast agents based on neural network model.

There is a need to standardize the process of micro/nanobubble preparation to bring it closer to clinical translation. We explored a neural network-based model to predict the structure-echogenicity relationship for the preparation and fabrication of ultrasound-enhanced contrast agents. Seven formulations were screened, and 109 measurements were obtained. An artificial neural network-multilayer perceptron (ANN-MLP) model was used. The original data were divided into the training and testing groups, which included 73 and 36 groups of data, respectively. The hidden layer was selected from three hidden layers and included bias. The classification graph showed that the predicted values of the training and testing groups were 76.7% and 66.7%, respectively. According to the receiver operating characteristic curve, the accuracy of different imaging effects could achieve a prediction rate of 88.1-96.5%. The percentage graph showed that the data were gradually converging. The predictive analysis curves of different ultrasound effects gradually approached stable value of Gain. Normalized importance predicted contributions for the Pk1, poly-dispersity index (PDI), and intensity account were 100%, 98.5%, and 89.7%, respectively. The application of the ANN-MLP model is feasible and effective for the exploration of the synthesis process of ultrasound contrast agents. 1,2-Distearoyl-sn-glycero-3 phosphoethanolamine-N (methoxy[polyethylene glycol]-2000) (DSPE PEG-2000) correlated highly with the success rate of contrast agent synthesis.

Pitfalls of Using Imaging Technique in the Presence of Eustachian Valve or Chiari Network: Effects on Right-to-Left Shunt and Related Influencing Factors.

When patent foramen ovale (PFO) combines with the prominent Eustachian valve or Chiari network (EV/CN), contrast transthoracic echocardiography (cTTE) may miss the diagnosis of PFO. We sought to determine the characteristics of right-to-left shunt (RLS) in PFO patients with prominent EV/CN on cTTE and identify the causal factors of missed diagnosis. We consecutively enrolled 98 patients who suffered from PFO-related stroke and with prominent EV/CN. All patients were divided into the delayed and non-delayed groups according to the characteristics of RLS on cTTE. The characteristics of RLS were compared with those of 42 intrapulmonary shunt patients. The anatomical characteristics of PFO and EV/CN were analyzed in the 98 PFO patients. Upon cTTE, significantly delayed occurrence and longer duration of the RLS in the delayed group were found both at rest and during the Valsalva maneuver, similar to the intrapulmonary shunt. Multivariate logistic analysis revealed that the length of EV/CN (>19 mm) and the diameter of PFO at the left atrium aspect (<1.2 mm) were high-risk factors for missed diagnosis. In conclusion, RLS showed delayed emergence and disappearance in some of the PFO patients with prominent EV/CN. The length of EV/CN and the diameter of PFO may have been related to the missed diagnosis of PFO.

The effect of gestational diabetes mellitus on fetal right heart growth in late-term pregnancy: A prospective study.

BACKGROUND: Gestational diabetes mellitus (GDM) is a complication of pregnancy strongly associated with an increased risk of structural fetal abnormalities. As the fetal heart grows quickly during the late-term pregnancy period, it is important to understand fetal heart growth before birth. This study explored how GDM affects fetal heart growth by evaluating basic echocardiography indicators during late pregnancy. METHODS: This prospective, longitudinal study included 63 GDM patients (GDM group) and 67 healthy pregnant women (control group). All subjects underwent fetal echocardiography scans at gestational weeks 28-32, 32-36, and 36-40. Twelve echocardiographic indicators were assessed at each observation and analyzed by using a mixed model. RESULTS: The left atrial diameter (LA) and left ventricular end-diastolic diameter (LV) similarly increased from the first to the third observation. The right ventricular end-diastolic diameter (RV) was significantly different between the groups, and a group × time interaction was detected. The tricuspid annular peak systolic velocity (s') increased more rapidly in the GDM than the control group during the first to second observations, and the group × time interaction was significant. The increase in the tricuspid annular plane systolic excursion (TAPSE) of the GDM group was "slow-fast", while that of the control group was "fast-slow", during three observations. After adjusting covariates, the group difference and interaction effect of TAPSE and RV remained significant. CONCLUSIONS: The differences in fetal right heart indicators between the GDM and control groups suggest that GDM may affect the structure and functional growth of the fetal right heart during late-term pregnancy.

Diagnosis of Atherosclerotic Plaques Using Vascular Endothelial Growth Factor Receptor-2 Targeting Antibody Nano-microbubble as Ultrasound Contrast Agent.

The atherosclerotic plaque is characterized by narrowing of blood vessels and reduced blood flow leading to the insufficient blood supply to the brain. The hemodynamic changes caused by arterial stenosis increase the shearing force of the fibrous cap on the surface of the plaque, thereby reducing the stability of the plaque. Unstable plaques are more likely to promote angiogenesis and increase the risk of patients with cerebrovascular diseases. A timely understanding of the formation and stability of the arterial plaque can guide in taking targeted measures for reducing the risk of acute stroke in patients. It has been confirmed that nano-microbubbles can enter these plaques through the gaps in the patient's vascular endothelial cells, thereby enhancing the acquisition of ultrasound information for plaque visualization. Therefore, we aim to investigate the diagnostic value of targeted nano-microbubbles for atherosclerotic plaques. This study constructed vascular endothelial growth factor receptor-2 (VEGFR-2) targeting antibody nano-microbubbles and compared its diagnostic value with that of blank nano-microbubbles for atherosclerotic plaques. Studies have found that VEGFR-2 targeting antibody nano-microbubbles can accurately detect the position of plaques. Its detection rate, sensitivity, and specificity for plaques are higher than those of blank nano-microbubbles. Similarly, the peak intensity and average transit time of VEGFR-2 targeting antibody nano-microbubbles were greater than those of blank nano-microbubbles. Therefore, we believe that the combination of VEGFR-2 antibody and nano-microbubbles can enhance the acquisition of ultrasound information on atherosclerotic plaque neovascularization, thereby improving the early diagnosis of unstable plaque.

Artificial Intelligence-Based Breast Cancer Diagnosis Using Ultrasound Images and Grid-Based Deep Feature Generator.

Purpose: Breast cancer is a prominent cancer type with high mortality. Early detection of breast cancer could serve to improve clinical outcomes. Ultrasonography is a digital imaging technique used to differentiate benign and malignant tumors. Several artificial intelligence techniques have been suggested in the literature for breast cancer detection using breast ultrasonography (BUS). Nowadays, particularly deep learning methods have been applied to biomedical images to achieve high classification performances. Patients and Methods: This work presents a new deep feature generation technique for breast cancer detection using BUS images. The widely known 16 pre-trained CNN models have been used in this framework as feature generators. In the feature generation phase, the used input image is divided into rows and columns, and these deep feature generators (pre-trained models) have applied to each row and column. Therefore, this method is called a grid-based deep feature generator. The proposed grid-based deep feature generator can calculate the error value of each deep feature generator, and then it selects the best three feature vectors as a final feature vector. In the feature selection phase, iterative neighborhood component analysis (INCA) chooses 980 features as an optimal number of features. Finally, these features are classified by using a deep neural network (DNN). Results: The developed grid-based deep feature generation-based image classification model reached 97.18% classification accuracy on the ultrasonic images for three classes, namely malignant, benign, and normal. Conclusion: The findings obviously denoted that the proposed grid deep feature generator and INCA-based feature selection model successfully classified breast ultrasonic images.

Evaluation of Achilles Tendon in Patients with Diabetic Foot Ulcer by High-Frequency Ultrasound Combined with Real-Time Shear-Wave Elastography.

To our best knowledge, no study investigated the utility of the combination of high-frequency US and real-time shear wave elastography (RTSWE) in the evaluation of Achilles tendon (AT). Our study aimed to evaluate AT in patients with diabetic foot ulcers using high-frequency US and RTSWE. We retrospectively reviewed 100 patients who visited our hospital due to diabetes. Patients were divided into the study group (those with diabetic foot ulcers) and the control group (those without diabetic foot ulcers). Patients' demographics, basic medical records, and laboratory tests were reviewed. High-frequency ultrasound and RTSWE were performed in both AT for all 100 patients. Young's modulus was measured at the upper, middle, and lower parts of each AT by RTSWE. There were 50 patients with diabetic foot ulcers. Patients in the study group had older age, a higher incidence of insulin use, and a higher level of cholesterol than those in the control group. However, thickness and Young's modulus of AT were comparable in two groups, when evaluated by the combination of high-frequency ultrasound and RTSWE. There was no significant difference in thickness and elastic modulus of AT in patients with and without diabetic foot ulcers when evaluated by the combination of high-frequency ultrasound and RTSWE.

Acceleration time to Ejection time ratio in fetal pulmonary artery system can predict neonatal respiratory disorders in gestational diabetic mellitus women.

BACKGROUND: Few researches studied fetal pulmonary pulse wave doppler and the clinical end point disorders in gestational diabetic mellitus (GDM) cohort. OBJECTIVE: To investigate fetal pulmonary artery acceleration time to ejection time ratio (PATET) in the prediction of neonatal respiratory disorders (NRD). METHODS: 238 pregnant women diagnosed with GDM who attended our hospital between February 2018 and March 2020 were retrospectively included. Fetal pulmonary artery Doppler wave measurements were recorded, including main, left and right pulmonary artery blood flow, and left and right peripheral pulmonary artery blood flow. Acceleration time (At)/ejection time (Et) were calculated. RESULTS: 183 GDM pregnant women and neonates were divided into NRD(+)(n = 42) and NRD(-) group (n = 141). 16 cases were neonatal pneumonia (NP) within 28 days after birth in NRD(+) group. The area under curve (AUC) of left peripheral pulmonary artery acceleration time to ejection time (LPPATET) was 0.967 (95% CI: 0.927∼1.000, P < 0.001) and the AUC of right peripheral pulmonary artery acceleration time to ejection time (RPPATET) was 0.967 (95% CI: 0.927∼1.000, P < 0.001), indicating that LPPATET and RPPATET are both predictors for NRD. The results of interobserver variabilty and intraobserver variability showed a good consistency. CONCLUSIONS: The At/Et of fetal peripheral pulmonary artery (PPA) in GDM women may be predictors for NRD, and the indicator can provide assistance in clinical management of diabetes pregnant patients.

Automated Analysis of Doppler Echocardiographic Videos as a Screening Tool for Valvular Heart Diseases.

OBJECTIVES: This study sought to develop a deep learning (DL) framework to automatically analyze echocardiographic videos for the presence of valvular heart diseases (VHDs). BACKGROUND: Although advances in DL have been applied to the interpretation of echocardiograms, such techniques have not been reported for interpretation of color Doppler videos for diagnosing VHDs. METHODS: The authors developed a 3-stage DL framework for automatic screening of echocardiographic videos for mitral stenosis (MS), mitral regurgitation (MR), aortic stenosis (AS), and aortic regurgitation (AR) that classifies echocardiographic views, detects the presence of VHDs, and, when present, quantifies key metrics related to VHD severities. The algorithm was trained (n = 1,335), validated (n = 311), and tested (n = 434) using retrospectively selected studies from 5 hospitals. A prospectively collected set of 1,374 consecutive echocardiograms served as a real-world test data set. RESULTS: Disease classification accuracy was high, with areas under the curve of 0.99 (95% CI: 0.97-0.99) for MS; 0.88 (95% CI: 0.86-0.90) for MR; 0.97 (95% CI: 0.95-0.99) for AS; and 0.90 (95% CI: 0.88-0.92) for AR in the prospective test data set. The limits of agreement (LOA) between the DL algorithm and physician estimates of metrics of valve lesion severities compared to the LOAs between 2 experienced physicians spanned from -0.60 to 0.77 cm2 vs -0.48 to 0.44 cm2 for MV area; from -0.27 to 0.25 vs -0.23 to 0.08 for MR jet area/left atrial area; from -0.86 to 0.52 m/s vs -0.48 to 0.54 m/s for peak aortic valve blood flow velocity (Vmax); from -10.6 to 9.5 mm Hg vs -10.2 to 4.9 mm Hg for average peak aortic valve gradient; and from -0.39 to 0.32 vs -0.31 to 0.32 for AR jet width/left ventricular outflow tract diameter. CONCLUSIONS: The proposed deep learning algorithm has the potential to automate and increase efficiency of the clinical workflow for screening echocardiographic images for the presence of VHDs and for quantifying metrics of disease severity.

[Tanshinone IIa attenuates vascular calcification through inhibition of NF-κB and ß-catenin signaling pathways].

Tanshinone IIa is a key ingredient extracted from the traditional Chinese medicine Salvia miltiorrhiza (Danshen), and is widely used to treat various cardiovascular diseases. Vascular calcification is a common pathological change of cardiovascular tissues in patients with chronic kidney disease, diabetes, hypertension and atherosclerosis. However, whether Tanshinone IIa inhibits vascular calcification and the underlying mechanisms remain largely unknown. This study aims to investigate whether Tanshinone IIa can inhibit vascular calcification using high phosphate-induced vascular smooth muscle cell and aortic ring calcification model, and high dose vitamin D3 (vD3)-induced mouse models of vascular calcification. Alizarin red staining and calcium quantitative assay showed that Tanshinone IIa significantly inhibited high phosphate-induced vascular smooth muscle cell and aortic ring calcification. qPCR and Western blot showed that Tanshinone IIa attenuated the osteogenic transition of vascular smooth muscle cells. In addition, Tanshinone IIa also significantly inhibited high dose vD3-induced mouse aortic calcification and aortic osteogenic transition. Mechanistically, Tanshinone IIa inhibited the activation of NF-κB and ß-catenin signaling in normal vascular smooth muscle cells. Similar to Tanshinone IIa, inhibition of NF-κB and ß-catenin signaling using the chemical inhibitors SC75741 and LF3 attenuated high phosphate-induced vascular smooth muscle cell calcification. These results suggest that Tanshinone IIa attenuates vascular calcification at least in part through inhibition of NF-κB and ß-catenin signaling, and Tanshinone IIa may be a potential drug for the treatment of vascular calcification.

KLK11 promotes the activation of mTOR and protein synthesis to facilitate cardiac hypertrophy.

BACKGROUND: Cardiovascular diseases have become the leading cause of death worldwide, and cardiac hypertrophy is the core mechanism underlying cardiac defect and heart failure. However, the underlying mechanisms of cardiac hypertrophy are not fully understood. Here we investigated the roles of Kallikrein 11 (KLK11) in cardiac hypertrophy. METHODS: Human and mouse hypertrophic heart tissues were used to determine the expression of KLK11 with quantitative real-time PCR and western blot. Mouse cardiac hypertrophy was induced by transverse aortic constriction (TAC), and cardiomyocyte hypertrophy was induced by angiotensin II. Cardiac function was analyzed by echocardiography. The signaling pathway was analyzed by western blot. Protein synthesis was monitored by the incorporation of [3H]-leucine. Gene expression was analyzed by quantitative real-time PCR. RESULTS: The mRNA and protein levels of KLK11 were upregulated in human hypertrophic hearts. We also induced cardiac hypertrophy in mice and observed the upregulation of KLK11 in hypertrophic hearts. Our in vitro experiments demonstrated that KLK11 overexpression promoted whereas KLK11 knockdown repressed cardiomyocytes hypertrophy induced by angiotensin II, as evidenced by cardiomyocyte size and the expression of hypertrophy-related fetal genes. Besides, we knocked down KLK11 expression in mouse hearts with adeno-associated virus 9. Knockdown of KLK11 in mouse hearts inhibited TAC-induced decline in fraction shortening and ejection fraction, reduced the increase in heart weight, cardiomyocyte size, and expression of hypertrophic fetal genes. We also observed that KLK11 promoted protein synthesis, the key feature of cardiomyocyte hypertrophy, by regulating the pivotal machines S6K1 and 4EBP1. Mechanism study demonstrated that KLK11 promoted the activation of AKT-mTOR signaling to promote S6K1 and 4EBP1 pathway and protein synthesis. Repression of mTOR with rapamycin blocked the effects of KLK11 on S6K1 and 4EBP1 as well as protein synthesis. Besides, rapamycin treatment blocked the roles of KLK11 in the regulation of cardiomyocyte hypertrophy. CONCLUSIONS: Our findings demonstrated that KLK11 promoted cardiomyocyte hypertrophy by activating AKT-mTOR signaling to promote protein synthesis.

Utility of lung ultrasound to identify interstitial lung disease: An observational study based on the STROBE guidelines.

ABSTRACT: Lung ultrasound (LUS) has recently been used to identify interstitial lung disease (ILD). However, data on the role of LUS in the detection of ILD remain limited. The aim of this study was to investigate the diagnostic value of LUS compared with high-resolution computed tomography (HRCT) in patients with ILD.The retrospective study was carried out by reviewing the medical records of patients with respiratory signs and symptoms discharged from the respiratory ward. Only patients with suspected ILD who underwent HRCT and LUS within a week were selected. ILD was identified with a semi-quantitative score of B-lines >5 and a Warrick score >0 points. The endpoints of LUS in diagnosing ILD (i.e., sensitivity, specificity, positive likelihood ratio [PLR], negative likelihood ratio [NLR], positive predictive value [PPV], and negative predictive value [NPV], and receiver operating characteristic [ROC] curve) was compared with that of HRCT. The reference standard used for the diagnosis of ILD was based on history, clinical findings and examination, and laboratory and instrumental tests, including pulmonary function tests, lung histopathology, and HRCT (without LUS findings).The final clinical diagnosis of ILD was 55 in 66 patients with suspected ILD. HRCT was positive in 55 patients, whereas LUS detected ILD in 51 patients. Four patients with negative LUS findings were positive on HRCT. The results showed 93% sensitivity, 73% specificity, 3.40 PLR, 0.10 NLR, 94% PPV, and 67% NPV for LUS, whereas 100% sensitivity, 82% specificity, 5.49 PLR, 0.01 NLR, 97% PPV, and 100% NPV for HRCT. Comparison of the 2 ROC curves revealed significant difference in the diagnostic value of the 2 methods for the diagnosis of ILD (P = .048).Our results indicated that LUS is a useful technique to identify ILD. Considering its non-radiation, portable and non-invasive advantages, LUS should be recommended as a valuable screening tool in patients with suspected ILD.