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.

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.

[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.

Very high contrast volume holographic gratings recorded in photopolymerizable nanocomposite materials.

Volume holographic phase gratings possessing the saturated refractive index modulation amplitudes as large as 4.5×10-2 were recorded at a wavelength of 532 nm in a photopolymerizable nanoparticle-polymer composite (NPC) film dispersed with ultrahigh refractive index hyperbranched-polymer (HBP) organic nanoparticles. This prominent result was achieved by a combination of the HBP nanoparticles with triazine and aromatic ring units and an electron donor/acceptor photo-initiator system doped in an acrylate monomer blend with low viscosity. As a result, efficient mutual diffusion of HBP nanoparticles and monomer having their very large refractive index difference took place. Obtained results suggest a potentiality of our newly developed HBP-dispersed NPC gratings as efficient volume holographic optical elements for various photonic applications including wearable headsets for augmented and mixed reality.

Assessment of left ventricular energy loss using vector flow mapping in patients with stages 1-3 chronic kidney disease.

BACKGROUND: Patients with chronic kidney disease (CKD) experience abnormality of intracardiac blood flow status during early-stages of disease. Left ventricular energy loss (EL) derived from vector flow mapping (VFM) represents fluid energy lost as heat in left ventricle and had been used to detect intracardiac blood flow efficiency. We aimed to evaluate the left ventricular EL in stage 1-3 CKD patients, and explored whether hypertension, a main cardiovascular risk, deteriorate the abnormality of intracardiac blood flow status. METHODS: Transthoracic echocardiography was performed in 41 controls and 48 patients with stages 1-3 CKD. CKD patients consisted a subgroup with no hypertension, a subgroup with well-controlled hypertension and a subgroup with poorly controlled hypertension. The EL were calculated in the left ventricle using VFM analysis from the apical 3-chamber view. Furthermore, the correlation and stepwise multiple regression analysis were used to explore the potential independent predictors of left ventricular EL. RESULTS: Compared with controls, stage 1-3 CKD patients showed increased left ventricular EL during total diastole, late diastole, total systole, isovolumic contraction and ejection. CKD patients with poorly controlled hypertension had higher left ventricular EL compared to the other CKD subgroups. Additionally, the ratio of mitral early filling wave peak velocity and early mitral annular peak velocity on septal side, mitral early filling wave peak velocity, and left ventricular mass index were independent predictors of the diastolic EL; whereas systolic blood pressure and left ventricular mass index were independent predictors of the systolic EL. CONCLUSIONS: Left ventricular EL was a useful echocardiographic parameter to evaluate the impaired intracardiac blood flow efficiency in patients with stages 1-3 CKD. Hypertension was a crucial contributor for intracardiac blood flow abnormality. This study might provide valuable clinical data to discern cardiac dysfunction and reduce the cardiovascular risk in early-stage CKD.

Evaluation of left ventricular diastolic function based on flow energetic parameters in chronic kidney disease with diastolic dysfunction.

BACKGROUND: Intra-ventricular blood flow dynamics is considered as an important component of left ventricular (LV) function assessment. The purpose of this study was to evaluate the LV diastolic function in chronic kidney disease (CKD) with different degrees of LV diastolic dysfunction (LVDD) by using flow energetic parameters. METHODS: In this study, a total of 96 cases were recruited, including 58 CKD patients and 38 healthy controls. CKD patients were divided into 2 groups according to LVDD severity, named as DD1 and DD2. Vector flow-mapping (VFM) analysis was executed to calculate left ventricle average energy loss (EL) during early filling phase (E-EL_ave), atrial filling phase (A-EL_ave), diastole phase (D-EL_ave), and ejection phase (S-EL_ave). Moreover, the average vortex circulation during early filling phase (E-cir_ave) and atrial filling phase (A-cir_ave) was also assessed in the apical three-chamber view. The rate of average EL during early filling and atrial filling was expressed as E/A-EL. RESULT: Compared to the control group, A-EL_ave, S-EL_ave, and A-cir_ave in the DD1 group were higher (P < 0.05), and all parameters were obviously higher in the DD2 group (P < 0.05). In the control group and the DD2 subgroup, the E-EL_ave value was significantly higher than A-EL_ave value, which was opposite to the DD1 group. As diastolic dysfunction worsened, E-EL_ave and D-EL_ave risen gradually (P < 0.05), and A-EL_ave and S-EL_ave were slightly elevated with no significance. There were significant correlations between LV diastolic function and flow energetic parameters. Stepwise multiple regression analysis revealed that various LV function parameters could be regarded as independent predictors of average diastolic EL (all P < 0.01). CONCLUSIONS: For CKD patient with LVDD and LVEF > 50%, effective LV filling and systolic ejection with optimized energy consumption have been impaired. As a new flow-derived index, EL can quantitatively evaluate LV diastolic function in terms of blood fluid dynamics in CKD with various LVDD.

Spatio-temporal image correlation rendering mode visualizes the specific location and surrounding structure of ventricular septal defect.

The limitations of traditional two-dimensional (2D) echocardiography in accurate diagnosis of ventricular septal defect (VSD) prompt the necessity of spatio-temporal image correlation (STIC) technology. Our study attemped to reconstruct the volume data of VSD using STIC rendering mode, and dynamically display the specific location and surrounding structure of VSD, in order to assist conventional 2D echocardiography. A total of 20 fetuses diagnosed as VSD using 2D echocardiography were enrolled in our study. Multiplane imaging mode was applied to obtain 2D four-chamber view, five-chamber view, and main artery short-axis view. STIC rendering mode was performed to collect volume data and visualize the shape, specific location and surrounding structure of VSD. A more detailed VSD classification for 20 VSD cases was identified. Based on the specific location and surrounding structure under STIC images, they were further subdivided into different types. STIC rendering mode intuitively showed the specific shape and surrounding structure of VSD. STIC technology can be used for definite classification and diagnosis of VSD, combining with 2D echocardiography to make a better therapeutic intervention in clinical practice. Clin. Anat. 32:408-420, 2019. © 2019 Wiley Periodicals, Inc.

The Association of Functional Capacity With Right Atrial Deformation in Patients With Pulmonary Arterial Hypertension: A Study With Two-Dimensional Speckle Tracking.

BACKGROUND: The purpose of this study was to assess right atrial (RA) myocardial mechanics in pulmonary hypertension (PH) patients using two-dimensional speckle tracking (2D-STE), and define the relationship between RA function and exercise capacity in PH patients. METHODS: Thirty-eight consecutive PH patients were studied and compared with a control group of 25 healthy volunteers. Peak atrial longitudinal strain (PALS), RA strain rate were measured in all subjects. PALS values were obtained by averaging all segments (global PALS), and by averaging segments measured in the four-chamber view. RESULTS: Right atrial PALS was significantly lower in PH patients than in controls, and gradually reduced with the development of cardiac insufficiency. A significantly positive correlation between global PALS and 6-minute walk distance (6MWD) was found (p=0.003). Furthermore, global PALS demonstrated the highest diagnostic accuracy (AUC of 0.979) and excellent sensitivity and specificity of 86.8% and 84%, respectively, to predict functional status using a cut-off value less than 38.08%. CONCLUSIONS: Right atrial deformation is significantly damaged in PH patients. Right atrial reservoir function can be estimated using 2D-STE, which gradually decreases in PH patients with different World Health Organization-functional class (WHO-FC). Right atrial PALS is a valuable factor for predicting the functional status and exercise capacity in PH patients.

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.

The incremental value of left ventricular energy loss in predicting adverse events in chronic kidney disease patients with preserved ejection fraction.

BACKGROUND: Vector flow mapping (VFM) is a new echocardiographic technology that can effectively evaluate systolic and diastolic hemodynamic function. However, little is known about the prognostic value of VFM-related parameters. In this paper we aimed to investigate whether left ventricular energy loss (EL) parameters as assessed by VFM enhance prediction of adverse events in patients with chronic kidney disease with preserved ejection fraction. METHODS: One hundred thirty-nine prospectively recruited patients (66% male, 58% on dialysis) with CKD stage 3-5 with normal left ventricular ejection fraction (LVEF) made up the study cohort. Global longitudinal strain (GLS) was calculated using 2-dimensional speckle tracking, and the LV EL during one cardiac cycle for each period was measured using VFM technology. Participants were followed for 4.17 ± 1.58 years for the primary end point of overall mortality and major adverse cardiovascular events (MACE). RESULTS: Forty-five (32%) patients had a primary endpoint event. The EL during each period especially during the ejection stage (Ej-EL) was significantly higher in patients with adverse events than in those without, meanwhile the LV GLS were lower. The Ej-EL (HR: 1.11; 95% CI: 1.06-1.15) and LV GLS (HR: 0.87; 95% CI: 0.81-0.94) (all P < .001) were independent predictors for the primary end point. Increased Ej-EL (≥6.13, 10-3 J/m s) and impaired GLS (<15.52, %) were associated with a higher risk of overall mortality death and MACE (log rank χ2 = 26.94, 7.19; P < .001, =0.007), and DeLong tests showed that Ej-EL (AUC = 0.823) has a slight advantage in predicting adverse events compared to GLS (AUC = 0.681). Furthermore, the addition of Ej-EL to a model with conventional parameters did more to improve the model's discrimination compared to GLS. CONCLUSIONS: Increased Ej-EL as determined by VFM is associated with a higher risk of overall death and MACE in CKD patients with preserved EF.

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.

Sirt7 protects against vascular calcification via modulation of reactive oxygen species and senescence of vascular smooth muscle cells.

Vascular calcification is frequently seen in patients with chronic kidney disease (CKD), and significantly increases cardiovascular mortality and morbidity. Sirt7, a NAD+-dependent histone deacetylases, plays a crucial role in cardiovascular disease. However, the role of Sirt7 in vascular calcification remains largely unknown. Using in vitro and in vivo models of vascular calcification, this study showed that Sirt7 expression was significantly reduced in calcified arteries from mice administered with high dose of vitamin D3 (vD3). We found that knockdown or inhibition of Sirt7 promoted vascular smooth muscle cell (VSMC), aortic ring and vascular calcification in mice, whereas overexpression of Sirt7 had opposite effects. Intriguingly, this protective effect of Sirt7 on vascular calcification is dependent on its deacetylase activity. Unexpectedly, Sirt7 did not alter the osteogenic transition of VSMCs. However, our RNA-seq and subsequent studies demonstrated that knockdown of Sirt7 in VSMCs resulted in increased intracellular reactive oxygen species (ROS) accumulation, and induced an Nrf-2 mediated oxidative stress response. Treatment with the ROS inhibitor N-acetylcysteine (NAC) significantly attenuated the inhibitory effect of Sirt7 on VSMC calcification. Furthermore, we found that knockdown of Sirt7 delayed cell cycle progression and accelerated cellular senescence of VSMCs. Taken together, our results indicate that Sirt7 regulates vascular calcification at least in part through modulation of ROS and cellular senescence of VSMCs. Sirt7 may be a potential therapeutic target for vascular calcification.

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.

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.

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 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.

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.

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.

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.