Purpose To prepare biomimetic targeted nanobubbles(NBs)named IR780-RBCM@NBs and explore the ability of performing ultrasound enhancement,escaping immune surveillance and targeting tumor cells.Materials and Methods The red blood cell membrane(RBCM)was extracted from mouse blood,the IR780-RBCM@NBs were prepared by modified membrane hydration method,the physical and chemical properties of nanobubbles were detected.In vitro,self-made equipment was used to test the contrast-enhanced ultrasound imaging ability of the nanobubbles.Laser confocal fluorescence microscopy was used to observe the characteristics of the nanobubbles retaining CD47 on the RBCM surface,and further to observe the phagocytosis of the nanobubbles by macrophages and the targeting ability of the nanobubbles to different tumor cells.Results The newly prepared IR780-RBCM@NBs had the appearance of emulsion,the particle size was about(522.4±58.6)nm and the dispersity was good.It was spherical in uniform size under the microscope.IR-780 could be detected on the nanobubbles,which had the dual-mode imaging capabilities of near-infrared fluorescence imaging and contrast-enhanced ultrasound imaging.The RBCM-specific protein CD47 could be detected on the surface of the nanobubbles.The nanobubbles exhibited excellent immune phagocytic escape function and high efficiency to target different tumor cells.Conclusion Newly prepared biomimetic targeted IR780-RBCM@NBs,with good performance,can escape immune phagocytosis and efficiently target tumors,providing a new idea for molecular targeted accurate diagnosis and treatment of tumors.

Objective:To evaluate the performance of the self-developed transcranial ultrasound patch probe as well as its stability and safety in clinical applications.Methods:This study was a cross-sectional study. Fifty healthy subjects in Tangdu Hospital from October to December 2023 were randomly and equally divided into two groups using a simple randomization method. The transcranial Doppler ultrasound (TCD) examinations were performed using the self-developed transcranial ultrasound patch probe and normal TCD probe alternatively in each group from squatting to sitting and then to standing posture. Two experienced sonographers (A, B) each completed one test for the same subject using the above two different probes. The differences of the ultrasound parameters of middle cerebral artery (MCA) blood flow measured by the self-developed transcranial ultrasound patch probe and the normal TCD probe in different postures (squatting, sitting, standing) were compared. At the same time, operator satisfaction with the use of the two probes was assessed in terms of time taken to obtain satisfactory images, operational comfort, and ease of handling. In addition, the subject's comfort during the examination was examined, and the signal acquisition stability of the two probes was comprehensively evaluated.Results:There was no significant difference in detection of various ultrasound parameters of MCA between using the transcranial patch probe and the normal TCD probe (all P>0.05). The overall satisfaction of transcranial ultrasound patch probe and normal TCD probe was quite similar. The transcranial ultrasound patch probe acquired favorable and stable signals. No subjects experienced adverse events/accidents during the examination of the subjects using the transcranial ultrasound patch probe as well as the normal TCD probe.Both probes had good and stable signal acquisition. Conclusions:TCD examinations could be well performed using the transcranial ultrasound patch probe, which might provide a potential new method for real-time monitoring and evaluating of cerebral blood flow of a moving subject.

Background: Gut microbiota is closely related to the occurrence and development of diabetes and affects the prognosis of diabetic complications, and the underlying mechanisms are only partially understood. We aimed to explore the possible link between the gut microbiota and vascular inflammation of diabetic mice. Methods: The db/db diabetic and wild-type (WT) mice were used in this study. We profiled gut microbiota and examined the and vascular function in both db/db group and WT group. Gut microbiota was analyzed by 16s rRNA sequencing. Vascular function was examined by ultrasonographic hemodynamics and histological staining. Clostridium butyricum (CB) was orally administered to diabetic mice by intragastric gavage every 2 days for 2 consecutive months. Reactive oxygen species (ROS) and expression of nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by fluorescence microscopy. The mRNA expression of inflammatory cytokines was tested by quantitative polymerase chain reaction. Results: Compared with WT mice, CB abundance was significantly decreased in the gut of db/db mice, together with compromised vascular function and activated inflammation in the arterial tissue. Meanwhile, ROS in the vascular tissue of db/db mice was also significantly increased. Oral administration of CB restored the protective microbiota, and protected the vascular function in the db/db mice via activating the Nrf2/HO-1 pathway. Conclusion This study identified the potential link between decreased CB abundance in gut microbiota and vascular inflammation in diabetes. Therapeutic delivery of CB by gut transplantation alleviates the vascular lesions of diabetes mellitus by activating the Nrf2/HO-1 pathway.

Objective:To explore the level of arterial stiffness and its influencing factors in prediabetic population.Methods:From June 2021 to June 2022, 207 prediabetes patients were prospectively and randomly recruited from the physical examination center and outpatient clinic of Tangdu Hospital of Air Force Military Medical University to be the prediabetic group and 130 healthy volunteers at the same time with the same gender and age as the healthy controls. The carotid-femoral pulse wave velocity (PWV), brachial-radial PWV, and femoral-ankle PWV were measured by an automatic ultrasonic arterial stiffness measurement technology. The common carotid artery wall intima-media thickness (IMT) and left heart function were routinely evaluated. A questionnaire was designed to investigate the subjects′ smoking, drinking, diet, staying up late, exercise and other living habits. Comparison between groups and multivariate linear regression analysis were used to analyze the relevant data.Results:The carotid-femoral PWV and common carotid artery wall IMT in prediabetic group were significantly higher than those in healthy controls [(7.10±2.00)m/s vs (6.26±1.14)m/s, (0.57±0.11)mm vs (0.51±0.08)mm; both P<0.001], but there were no significant differences in the brachial-radial PWV and femoral-ankle PWV between the two groups (both P>0.05). Multivariate linear regression analysis showed that prediabetes was an independent influencing factor in carotid-femoral PWV after adjusting for confounding factors ( P<0.001), in addition, age ( P<0.001), diastolic blood pressure ( P<0.001), staying up late ( P=0.011) and low density lipoprotein cholesterol ( P=0.022) were also the independent influencing factors of carotid-femoral PWV. Conclusions:Compared with healthy people, the stiffness of aorta is significantly increased in prediabetic people, but there is no significant change in the stiffness of peripheral arteries. Prediabetes, age, diastolic blood pressure, staying up late and low density lipoprotein cholesterol are independent influencing factors of carotid-femoral PWV.

Objective:To investigate the effect of self-made phase-change nanodroplets IR780/FA-Nds-DTX as molecular targeted ultrasound contrast agents for accurate diagnosis and combined targeted therapy of pancreatic cancer.Methods:Pancreatic cancer cell lines were cultured in vitro and 50 tumor bearing nude mouse models were established. The experimental group (IR780/ FA-NdS-DTX) and four control groups[ normal saline, Nds(FITC), FA-Nds (FITC) and IR-780] were divided to verify dual-mode targeted imaging. The imaging using the IVIS Imaging System verified the high-efficiency targeted detection ability and near-infrared fluorescence imaging of IR780/FA-Nds-DTX for tumors in vivo, phase transformation induced by low-intensity focused ultrasound and further contrast-enhanced ultrasound imaging verified the high-efficiency aggregation of IR780/FA-Nds-DTX in local tumors and accurate evaluation of tumor contour. The therapeutic effect was observed in the experimental group (IR780/FA-Nds-DTX) and four control groups (FA-Nds-IR780, FA-Nds-DTX, FA-Nds and normal saline). After low-intensity focused ultrasound irradiation for 30s induced microbubble blasting after phase transformation in each group, 808nm photothermal therapy apparatus was used to irradiate tumor area in each group. Two-dimensional ultrasound was used to monitor the changes in tumor volume in each group before and at 3 d, 9 d, 15 d after treatment, and the changes in tumor volume rate and inhibition in each group were statistically analyzed and compared.Results:The amount of IR780/ FA-Nds-DTX locally targeted aggregation was the largest, and the average fluorescence intensity of tumor in the experimental group was significantly higher than that of the control groups: IR780/ FA-Nds-DTX group compared with Nds(FITC) group[(5.12±0.69)×10 7 vs (1.06±0.23)×10 7, P<0.05], IR780/FA-Nds-DTX group compared with FA-Nds (FITC) group [(5.12±0.69)×10 7 vs (2.98±0.34)×10 7, P<0.05], IR780/FA-Nds-DTX group compared with IR-780 group [(5.12±0.69)×10 7 vs (1.54±0.42)×10 7, P<0.05], and there was no fluorescence in tumor area in saline group. Further contrast-enhanced ultrasound imaging after nanodroplet phase transformation could more accurately locate the tumor boundary. After 15 days of photothermal ablation combined with chemotherapy, the growth rate of tumor volume in the IR780/ FA-Nds-DTX treatment group was significantly lower than that in the control groups: IR780/FA-Nds-DTX group compared with FA-Nds-IR780 group[(0.105±0.075) vs (0.405±0.175), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds-DTX group [(0.105±0.075) vs (1.385±0.035), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds group [(0.105±0.075) vs (2.255±0.105), P<0.05], IR780/ FA-Nds-DTX group compared with normal saline group [(0.105±0.075) vs (2.185±0.155), P<0.05]. And the tumor inhibition rate increased significantly: IR780/ FA-Nds-DTX group compared with FA-Nds-IR780 group [(0.93±0.06) vs (0.48±0.17), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds-DTX group [(0.93±0.06) vs (-0.51±0.105), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds group [(0.93±0.06) vs (-1.63±0.115), P<0.05], IR780/ FA-Nds-DTX group compared with normal saline[(0.93±0.06) vs (-1.35±0.245), P<0.05]. Conclusions:The self-made phase-change ultrasound contrast agents IR780/FA-Nds-DTX have good potential clinical value in targeted detection and combined therapy of pancreatic cancer with small lesions or even metastases.

Objective:To investigate the application of ultrasound imaging in evaluating the curative effect of patients with nutcracker syndrome (NCS) treated with three-dimensional(3D) printing extravascular titanium stents.Methods:From December 2015 to December 2018, a total of 41 NCS patients enrolled in the Second Affiliated Hospital of the Air Force Military Medical University to receive 3D-printing extravascular titanium stents treatment were included in this study. Ultrasound and CT angiography (CTA) were performed before and 1 week after operation to evaluate the compression of left renal vein and related hemodynamic changes.Results:Before the operative, the ultrasound examination showed that the angle between the abdominal aorta and superior mesenteric artery was (21.29±4.53)°, which was significantly improved in 1 week after treatment [(47.42±7.45)°, P<0.001]. Left renal vein was significantly compressed before treatment, with the smallest inner diameter of (1.51±0.49)mm and its peak blood flow velocity was (143.92±50.40)cm/s. Postoperative ultrasound images showed the significant expansion of left renal vein and no high velocity blood flow was observed. Ultrasound examination also showed that the diameter of left renal vein at renal hilum, blood flow velocity at renal hilum and inner diameter of spermatic vein were significantly improved after treatment ( P<0.001). Similar to ultrasound results, CTA also found that the angle between abdominal aorta and superior mesenteric artery increased significantly after treatment[(17.59±4.56)° vs (52.27±9.01)°, P<0.001]. CTA also showed the left renal vein was compressed before treatment, with the smallest inner diameter of (2.09±0.86)mm, and it was significantly expanded after operation. The inner diameter of left renal vein at renal hilum was also significantly reduced after treatment ( P<0.001). Conclusions:Ultrasound imaging can effectively evaluate the LRV compression and related hemodynamic changes before and after 3D printed stent implantation treatment in NCS patients and provide a reliable method to evaluate its clinical efficacy for these patients.

Objective:To observe and evaluate the imaging of the substantia nigra (SN) in the midbrain of 6-hydroxydopamine(6-OHDA)-constructed PD rat model by transcranial sonography (TCS), and to observe whether the substantia nigra hyperechogenicity (SNH) in nigral region appeared stably, for the purpose of providing a scientific method and theoretical basis for the selection and construction of laboratory animal disease model of PD.Methods:Rats were divided into vehicle group and 6-OHDA group, and received stereotactic injection. After 15 days, TCS was performed to observe both model and vehicle group whether the characteristic SNH appeared in ipsilateral substantia nigra compared with contralateral. The rats were sacrificed after detection, and the number of dopaminergic (DA) neurons and fibers in the nigrostriatal pathway of each rat were observed by immunofluorescence staining. Protein expression levels were observed by western blot. These experimental results of the 6-OHDA group and the vehicle group were compared to evaluate whether the model was successfully constructed and compared with the performance of their ultrasound images.Results:Nine rats survived PD model group and vehicle group. 6-OHDA PD rat models appeared stable SNH under the TCS examination, and the average area of SNH was (3.258±0.220)cm 2. Immunofluorescence staining showed apparently loss of TH-ir cells; Western blot showed TH protein level of ipsilateral dramatically decreased. Conclusions:The SNH consistently emerges in 6-OHDA unilateral rat PD models with relatively high survival rate and success rate.6-OHDA rat models can be used to study the unclear mechanism of SNH.

Objective:To prepare a dual targeting nanobubble both for HER2 target and cancer cell target, and compare the targeting ability with single targeting nanobubble either with HER2 targeting or cancer cell targeting ability in vitro.Methods:Nanobubbles(NBs) were prepared using the modified thin film hydration method and then connected with IR783 directly (NBs-IR783), HER2 antibody Affibody by avidin-biotin method (NBs-Affibody), or both of them (IR783-NBs-Affibody). The size distribution, stability, and biosafety of these contrast agents were observed. Flow cytometry and immunofluorescence were applied to compare the binding rate of these NBs against HER2 positive breast cancer cell in vitro.Results:The average size of NBs-Affibody, NBs-IR783 and IR783-NBs-Affibody was (538.4±95.8)nm, (551.8±114.8)nm, and (482.7±54.3)nm, respectively. The binding rate for HER2 positive cell was 26.6%, 97.6%, 84.5%, while for HER2 negative cell was 5.4%, 99.9% and 99.3%, respectively. The results of dual-labeled flow cytometry showed that the binding rate of IR783-NBs-Affibody mediated by IR783 to HER2 positive breast cancer cells were 79.5% and Affibody mediated HER2 targeting binding rate were 19.4%. However, NBs-IR783 only showed IR783-mediated tumor targeted binding without HER2 targeting ability with binding rate of 2.3%.Conclusions:The prepared IR783-NBs-Affibody has the dual targeting ability for HER2 positive breast cancer cell, which is superior to the single targeting NBs (NBs-Affibody and NBs-IR783). IR783-NBs-Affibody is optimal to meet the imaging requirements of tumor heterogeneity.

To evaluate the accuracy of transcranial color‐code sonography ( TCCS) in non‐invasive assessment of intracranial pressure( ICP ) . TCCS was used to monitor the cerebral hemodynamic parameters of patients with acute severe traumatic brain injury after decompressive craniectomy and make estimation of the non‐invasive intracranial pressure ( ICPtccs) . Methods A total of 91 patients with acute severe traumatic brain injury involved in this retrospective study were divided into the ICP normal group( ≤22 mm Hg ) and the ICP increased group ( ＞22 mm Hg ) . T he correlation and consistency of middle cerebral artery blood flow parameters and ICPtccs with invasive intracranial pressure ( iICP ) were analyzed . According to Glasgow score ( GCS) ,Patients( GCS 3－8) were divided into acute extremely severe traumatic brain injury( GCS 3 －5) and acute severe traumatic brain injury ( GCS 6 －8 ) . A comparison was made of ROC ( ICPtccs) curve and the area under the curve( AUC) between the two groups were cornpared . Results①No statistical differences were found in cerebral hemodynamic parameters between the side with and without decompressive craniectomy in patients with acute severe traumatic brain injury ( all P ＞0 .05 ) . ②M onitored resistive index ( RI) ,pulsatility index ( PI) and ICPtccs between the normal ICP group and the increased ICP group showed statistically significant differences ( all P ＜ 0 .05 ) ,w hile monitored systolic velocity ,diastolic velocity and mean velocity presented no statistically significant difference ( all P ＞0 .05) . T he correlations between RI ,PI with iICP were low ( r＝ 0 .247 ,0 .221 ; all P ＜ 0 .05 ) ,w hile there was a moderate correlation between ICPtccs and iICP( r ＝0 .417 , P ＜0 .001 ) . ③Bland‐Altman plot showed an overestimation of 2 .3 mm Hg ( 95% CI 0 .00－4 .59 mm Hg ) for ICPtccs compared to iICP . ④T he AUC of Glasgow score ( GCS 3－5 and GCS 6－8) in the two groups were 0 .759 ,0 .781 ( all P ＜0 .05) . All the cut‐off points of ICPtccs were 19 mm Hg ,with a sensitivity of 83 .33% ,81 .82% and a specificity of 64 .86% , 75 .68% ,respectively . Pairwise comparison of two AUCs showed no statistical difference ( P ＝ 0 .476) . ICPtccs presented the same ability to estimate ICP in patients with acute severe and extremely severe traumatic brain injury . TCCS could accurately assess the elevation of ICP in 72 .52% patients with acute severe traumatic brain injury . Conclusions TCCS can be used as a non‐invasive screening tool to assess w hether ICP of patients with acute severe traumatic brain injury is elevated and to semi‐quantitatively estimate ICP ,showing useful clinical value .

Objective To observe the blood flow changes of left ventricular cavity and quantify the energy loss (EL) and circulation of left ventricular during systole and diastole in patients with dilated cardiomyopathy(DCM) via vector flow mapping(VFM).Methods Thirty-six healthy volunteers and 32DCM patients were enrolled.According to the severity of mitral regurgitation(MR),DCM patients were divided into two groups.The quantitative parameters,including average energy loss(EL-base,EL mid,EL-apex) and circulation (vortex quantity,vortex area,circulation) were measured in the different periods of VFM imaging mode in apical four-chamber view,apical three-chamber view and apical two-chamber view respectively.The difference of parameters was evaluated between different groups during the different periods.The early transmitral valve blood flow velocity E,the late transmitral valve blood flow velocity A,the ratio of early transmitral valve blood flow velocity to the early diastolic mitral valve velocity E/e,the ratio of early transmitral valve blood flow velocity to the late transmitral valve blood flow velocity E/A and Tei index were derived via dual-Doppler imaging technology.Meanwhile,the correlation between VFM parameters with E,A,E/A,E/e or Tei index was also analyzed.Results ①The level of EL were decreased significantly among EL-mid and EL-apex during early diastole,EL-mid during mid-systole,EL-mid and EL-apex during late-systole (P ＜0.05),when compared with the control group.The vortex quantity,vortex area and circulation during early diastole and the vortex area and circulation during atrial contraction period were significantly different between two groups (all P ＜ 0.05).As for mid-diastole,only vortex area showed significant difference (P ＜0).01).(②) When compared with different grades of MR in DCM patients,the level of all segments EL during diastole and EL-base during systole increased with severity of MR(P ＜0.05).The circulation during diastole also increased with the severity of MR(P ＜0.05).③The vortex area and circulation had positive correlation with E/e during diastole in all subjects(all P ＜0.05).The vortex area during atrial contraction period and early diastole had positive correlation with E/A and Tei index in all subjects(r =0.630,0.345,0.468,0.316,0.663,0.264,respectively,allP ＜0.01).EL-mid during late-systole were negatively correlated with LV end diastolic volume,end systolic volume,but positively correlated with ejection fraction in all subjects(r =0.335,-0.312,0.340,respectively,all P ＜0.05).While,there was positive correlation between E/A with the basal segment of mid-diastole(r =0.376,P ＜0.01).Conclusions The visualization of hydromechanics inside the heart of DCM patients could be directly accomplished by VFM,which can provide a new method for diagnosis and evaluation of DCM.