Mx250 probe

Contrast enhanced US was performed using a Vevo3100 scanner (FUJIFILM Visualsonics, Joop Geesinkweg 140, 1114 AB Amsterdam, Netherlandsusing) with a MX250 probe (Centre Transmit Frequency: 20 MHz, Axial Resolution: 75 μm). SV (50 µL) was administered i.v. to anaesthetised mice while imaging was acquired in Non-Linear Contrast mode. A second injection of SV was administered 10 min later and high amplitude US bursts were applied every 10 s in order to acquire destruction-replenishment curves and study the perfusion kinetics (see SI). VevoLab software was used to trace regions of interest within the tumour and quantify contrast intensity as a function of time.

After treatment with HJ11 decoction for 28 days, rats of each group were subjected to echocardiographic analysis using Vevo3100 with an MX250 probe (FUJIFILM VisualSonics, Inc., Toronto, ON, Canada) at an ultrasound frequency of 21 MHz. Before echocardiographic analysis, the rats were anesthetized by inhalation of 2% isoflurane and placed in the supine position on a thermal blanket at 37°C. Long-axis B-ultrasound and M-ultrasound images of the heart were obtained by performing ultrasound examination in the left parasternal region. Then, the MX250 probe was rotated 90° to acquire short-axis B-ultrasound and M-ultrasound images at the papillary muscle level. Short-axis M-ultrasound images of the left ventricular region were performed to measure the left ventricular end-diastolic diameter (LVEDd), left ventricular ejection fraction (LVEF), left ventricular end-systolic diameter (LVEDs), and left ventricular fractional shortening (LVFS) of rats. All ultrasound measurements were performed over three consecutive cardiac cycles to calculate the average values.

FUJIFILM Vevo 3100LT system and MX250 probe were used to detect ultrasound. Pre‐detection chest hair removal was performed to expose the overall chest skin. The mice were anesthetized (maintenance oxygen flow 2.0/min, isoflurane 1.5%–2.0%), maintained at 38.0°C, and their heart rate was 400–500 beats per minute. The left ventricular long‐axis activity images were obtained in Parasternal long‐axis views in B‐Mode, and the mitral flow spectra were obtained in pulse‐wave mode in the apical four chambers at the maximum opening of the mitral valve. An ultrasound analysis was performed using Vevo LAB 3.2.6, and an average of at least five cycles of systolic and diastolic function was recorded.