Perceived stress mediates the association between perceived control and emotional distress: The moderating role of psychological resources and sex differences.

Studies have confirmed that perceived control is strongly negatively correlated with emotional distress. However, few studies have explored whether perceived stress plays a potential mediating role in this relationship and whether the association between perceived stress and emotional distress is moderated by psychological resources, such as self-esteem and social support. Furthermore, it is unclear whether there are sex differences in the moderating effects of psychological resources on emotional distress. A total of 951 healthy adults (51.84% females) from different regions of mainland China participated in the study and completed questionnaires in early December 2022, when prevention and control policies concerning COVID-19 in China underwent rapid change. Perceived control negatively correlated with emotional distress, and perceived stress mediated the association between perceived control and emotional distress. In addition, both internal (i.e., self-esteem) and external psychological resources (i.e., social support) moderated the association between perceived stress and emotional distress, and the positive correlation between perceived stress and emotional distress was higher in individuals with low social support (and self-esteem) than in those with high social support (and self-esteem). We found sex differences in the moderating roles of psychological resources. Specifically, self-esteem had a moderating effect on both men and women, whereas social support had a moderating effect only on women. These findings improve understanding of the relationship between perceived control and emotional distress and suggest that intervention programs should be designed to target men and women differently.

Biosynthetic Melanin/Ce6-Based Photothermal and Sonodynamic Therapies Significantly Improved the Anti-Tumor Efficacy.

Photothermal therapy (PTT) and sonodynamic therapy (SDT) are becoming promising therapeutic modalities against various tumors in recent years. However, the single therapeutic modality with SDT or PTT makes it difficult to achieve a satisfactory anti-tumor outcome due to their own inherent limitations, such as poor tissue penetration for the near-infrared (NIR) laser and the limited cytotoxic reactive oxygen species (ROS) generated from conventional sonosensitizers irradiated by ultrasound (US). Here, we successfully biosynthesized melanin with a controllable particle size with genetically engineered bacteria harboring a heat-inducible gene circuit. The biosynthetic melanin with 8 nm size and chlorin e6 (Ce6) was further encapsulated into liposomes and obtained SDT/PTT dual-functional liposomes (designated as MC@Lip). The resulting MC@Lip had an approximately 100 nm particle size, with 74.71% ± 0.54% of encapsulation efficiency for melanin and 94.52% ± 0.78% for Ce6. MC@Lip exhibited efficient 1O2 production and photothermal conversion capability upon receiving irradiation by US and NIR laser, producing significantly enhanced anti-tumor efficacy in vitro and in vivo. Especially, US and NIR laser irradiation of tumors received with MC@Lip lead to complete tumor regression in all tested tumor-bearing mice, indicating the great advantage of the combined use of SDT and PTT. More importantly, MC@Lip possessed good photoacoustic (PA) and fluorescence dual-modal imaging performance, making it possible to treat tumors under imaging guidance. Our study provides a novel approach to synthesize a melanin nanoparticle with controllable size and develops a promising combined SDT/PTT strategy to treat tumors.

Remote control of the recruitment and capture of endogenous stem cells by ultrasound for in situ repair of bone defects.

Stem cell-based tissue engineering has provided a promising platform for repairing of bone defects. However, the use of exogenous bone marrow mesenchymal stem cells (BMSCs) still faces many challenges such as limited sources and potential risks. It is important to develop new approach to effectively recruit endogenous BMSCs and capture them for in situ bone regeneration. Here, we designed an acoustically responsive scaffold (ARS) and embedded it into SDF-1/BMP-2 loaded hydrogel to obtain biomimetic hydrogel scaffold complexes (BSC). The SDF-1/BMP-2 cytokines can be released on demand from the BSC implanted into the defected bone via pulsed ultrasound (p-US) irradiation at optimized acoustic parameters, recruiting the endogenous BMSCs to the bone defected or BSC site. Accompanied by the daily p-US irradiation for 14 days, the alginate hydrogel was degraded, resulting in the exposure of ARS to these recruited host stem cells. Then another set of sinusoidal continuous wave ultrasound (s-US) irradiation was applied to excite the ARS intrinsic resonance, forming highly localized acoustic field around its surface and generating enhanced acoustic trapping force, by which these recruited endogenous stem cells would be captured on the scaffold, greatly promoting them to adhesively grow for in situ bone tissue regeneration. Our study provides a novel and effective strategy for in situ bone defect repairing through acoustically manipulating endogenous BMSCs.

Ultrasound Molecular Imaging of Neovascularization for Evaluation of Endometrial Receptivity Using Magnetic iRGD-Modified Lipid-Polymer Hybrid Microbubbles.

Background: Angiogenesis plays an important role in endometrial receptivity, determining the response of the endometrium to the blastocyst at the early stage of embryo implantation. During the application of assisted reproduction technologies, it is very important to evaluate the status of uterine angiogenesis before deciding on embryo implantation. Targeted microbubbles (MBs)-based ultrasound molecular imaging (UMI) can noninvasively detect the expression status of biomarkers at the molecular level, thereby being a potential diagnosis strategy for various diseases and their therapeutic evaluation. Methods: The iRGD-lipopeptide (DSPE-PEG2000-iRGD) conjugate was prepared with iRGD peptides and DSPE-PEG2000-maleimide through the Michael-type addition reaction. Then, the magnetic iRGD-modified lipid-polymer hybrid MBs (Mag-iLPMs) were prepared with the double-emulsification-solvent-evaporation method. Magnetic targeting of Mag-iLPMs was confirmed under the microscope, followed by a rectangular magnet. Next, the in vitro targeted binding of MBs to murine brain-derived endothelial cells.3 (bEnd.3) and human umbilical vein endothelial cells (HUVEC) were evaluated. The ratio of MBs binding to bEnd.3 and HUVEC at the same field was also compared. For in vivo studies, bolus injections of targeted or control MBs were randomly administrated to non-pregnant or pregnant rats on day 5. Then, the uteri were imaged using a VisualSonics Vevo 2100 ultrasound system (Fujifilm VisualSonics Inc., Ontario, Canada) equipped with a high-frequency transducer. Ultrasonic imaging signals were acquired from Mag-iLPMs, and compared with Mag-LPMs, iLPMs, and LPMs. Results: The Mag-iLPMs showed excellent performance in ultrasound contrast imaging and binding affinity to target cells. Using the magnetic field, 10.5- and 12.47-fold higher binding efficiency to bEnd.3 and HUVEC were achieved compared to non-magnetic iLPMs, respectively. Significantly enhanced UMI signals were also observed in the uteri of rats intravenously injected pregnant rats (6.58-fold higher than rats injected with iLPMs). Conclusion: We provided a powerful ultrasonic molecular functional imaging tool for uterine angiogenesis evaluation before embryonic implantation.

An Artificial Intelligence Model Based on ACR TI-RADS Characteristics for US Diagnosis of Thyroid Nodules.

Background US-based diagnosis of thyroid nodules is subjective and influenced by radiologists' experience levels. Purpose To develop an artificial intelligence model based on American College of Radiology Thyroid Imaging Reporting and Data System characteristics for diagnosing thyroid nodules and identifying nodule characteristics (hereafter, MTI-RADS) and to compare the performance of MTI-RADS, radiologists, and a model trained on benign and malignant status based on surgical histopathologic analysis (hereafter, MDiag). Materials and Methods In this retrospective study, 1588 surgically proven nodules from 636 consecutive patients (mean age, 49 years ± 14 [SD]; 485 women) were included. MTI-RADS and MDiag were trained on US images of 1345 nodules (January 2018 to December 2019). The performance of MTI-RADS was compared with that of MDiag and radiologists with different experience levels on the test data set (243 nodules, January 2019 to December 2019) with the DeLong method and McNemar test. Results The area under the receiver operating characteristic curve (AUC) and sensitivity of MTI-RADS were 0.91 and 83% (55 of 66 nodules), respectively, which were not significantly different from those of experienced radiologists (0.93 [P = .45] and 92% [61 of 66 nodules; P = .07]) and exceeded those of junior radiologists (0.78 [P < .001] and 70% [46 of 66 nodules; P = .04]). The specificity of MTI-RADS (87% [154 of 177 nodules]) was higher than that of both experienced and junior radiologists (80% [141 of 177 nodules; P = .02] and 75% [133 of 177 nodules; P = .001], respectively). The AUC of MTI-RADS was higher than that of MDiag (0.91 vs 0.84, respectively; P = .001). In the test set of 243 nodules, the consistency rates between MTI-RADS and the experienced group were higher than those between MTI-RADS and the junior group for composition (79% [n = 193] vs 73% [n = 178], respectively; P = .02), echogenicity (75% [n = 183] vs 68% [n = 166]; P = .04), shape (93% [n = 227] vs 88% [n = 215]; P = .04), and smooth or ill-defined margin (72% [n = 174] vs 63% [n = 152]; P = .002). Conclusion The area under the receiver operating characteristic curve (AUC) of an artificial intelligence model based on the American College of Radiology Thyroid Imaging Reporting and Data System (TI-RADS) was higher than that of a model trained on benign and malignant status based on surgical histopathologic analysis. The AUC and sensitivity of the model based on TI-RADS exceeded those of junior radiologists; the specificity of the model was higher than that of both experienced and junior radiologists. © RSNA, 2022.

Ultrasound imaging tracking of mesenchymal stem cells intracellularly labeled with biosynthetic gas vesicles for treatment of rheumatoid arthritis.

Rationale: Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation and damage to articular tissues that can lead to irreversible joint damage and progressive disability. The multipotent mesenchymal stem cells (MSCs) play an important role in immune disorders and tissue regeneration. However, their immunosuppressive effects and the underlying mechanisms are largely unclear due to the lack of tools for real-time imaging of MSCs in vivo. Gas vesicles (GVs) are biosynthetic nanobubbles that are ejected from aquatic microbes, such as bacteria and archaea, and have an excellent ultrasound imaging capacity. Methods: We harvested MSCs from the bone marrow of Sprague Dawley (SD) rats. Then, GVs were synthesized and incubated with MSCs to obtain intracellularly labeled MSCs. We firstly tested the ultrasound imaging of GV@MSCs in vitro and in vivo and then explored the therapeutic effect of GV@MSCs combined with methotrexate (MTX) in RA rats. Results: These GV@MSCs showed significant contrast-enhanced ultrasound signals without a loss of viability and differentiation capacity. In addition, the GV@MSCs could be imaged in real-time for 5 days using ultrasound both in vitro and in vivo, making it possible to visually track their migration and homing to the joint cavity from the subcutaneous layer of lateral malleolus joints in the injected RA rats. Furthermore, GV@MSCs significantly enhanced the curative effect of methotrexate (MTX) against RA, resulting in decreased paw thickness, lower arthritis index score, reduced bone erosion and cartilage destruction, compared to the PBS, free MTX, and GV@MSCs groups. Conclusion: We developed a novel therapeutic strategy against RA using GVs-loaded MSCs that can be tracked in vivo in real-time.

The LPS induced pyroptosis exacerbates BMPR2 signaling deficiency to potentiate SLE-PAH.

Pulmonary arterial hypertension (PAH) is a common and fatal complication of systemic lupus erythematosus (SLE). Whether the BMP receptor deficiency found in the genetic form of PAH is also involved in SLE-PAH patients remains to be identified. In this study, we employed patient-derived samples from SLE-associated PAH (SLE-PAH) and established comparable mouse models to clarify the role of BMP signaling in the pathobiology of SLE-PAH. Firstly, serum levels of LPS and autoantibodies (auto-Abs) directed at BMP receptors were significantly increased in patients with SLE-PAH compared with control subjects, measured by ELISA. Mass cytometry was applied to compare peripheral blood leukocyte phenotype in patients prior to and after treatment with steroids, which demonstrated inflammatory cells alteration in SLE-PAH. Furthermore, BMPR2 signaling and pyroptotic factors were examined in human pulmonary arterial endothelial cells (PAECs) in response to LPS stimulation. Interleukin-8 (IL-8) and E-selectin (SELE) expressions were up-regulated in autologous BMPR2+/R899X endothelial cells and siBMPR2-interfered PAECs. A SLE-PH model was established in mice induced with pristane and hypoxia. Moreover, the combination of endothelial specific BMPR2 knockout in SLE mice exacerbated pulmonary hypertension. Pyroptotic factors including gasdermin D (GSDMD) were elevated in the lungs of SLE-PH mice, and the pyroptotic effects of serum samples isolated from SLE-PAH patients on PAECs were analyzed. BMPR2 signaling upregulator (BUR1) showed anti-pyroptotic effects in SLE-PH mice and PAECs. Our results implied that deficiencies of BMPR2 signaling and proinflammatory factors together contribute to the development of PAH in SLE.

The value of the Demetics ultrasound-assisted diagnosis system in the differential diagnosis of benign from malignant thyroid nodules and analysis of the influencing factors.

OBJECTIVES: To evaluate the value of Demetics and to explore whether Demetics can help radiologists with varying years of experience in the differential diagnosis of benign from malignant thyroid nodules. METHODS: The clinical application value of Demetics was assessed by comparing the diagnostic accuracy of radiologists before and after applying Demetics. This retrospective analysis included 284 thyroid nodules that underwent pathological examinations. Two different combined methods were applied. Using method 1: the original TI-RADS classification was forcibly upgraded or downgraded by one level when Demetics classified the thyroid nodules as malignant or benign. Using method 2: the TI-RADS and benign or malignant classification of the thyroid nodules were flexibly adjusted after the physician learned the Demetics' results. RESULTS: Demetics exhibited a higher sensitivity than did junior radiologist 1 (pD1 = 0.029) and was similar in sensitivity to the two senior radiologists. Demetics had a higher AUC than both junior radiologists (pD1 = 0.042, pD2 = 0.038) and an AUC similar to that of the senior radiologists. The sensitivity (p = 0.035) and AUC (p = 0.031) of junior radiologist 1 and the specificity (p < 0.001) and AUC (p = 0.026) of junior radiologist 2 improved with combined method 1. The AUC of junior radiologist 2 improved with combined method 2 (p = 0.045). The factors influencing the diagnostic results of Demetics include sonographic signs (echogenicity and echogenic foci), contrast of the image, and nodule size. CONCLUSION: Demetics exhibited high sensitivity and accuracy in the differential diagnosis of benign from malignant thyroid nodules. Demetics could improve the diagnostic accuracy of junior radiologists. KEY POINTS: • Demetics exhibited a high sensitivity and accuracy in the differential diagnosis of benign from malignant thyroid nodules. • Demetics could improve the diagnostic accuracy of junior radiologists in the differential diagnosis of benign from malignant thyroid nodules. • Factors influencing the diagnostic results of Demetics include the sonographic signs (echogenicity and echogenic foci), contrast of the image, and nodule size.

Transvaginal four-dimensional hysterosalpingo-contrast sonography: Pain perception and factors influencing pain severity.

AIM: The purpose is to investigate pain perception during transvaginal four-dimensional hysterosalpingo-contrast sonography (TV 4D-HyCoSy) and factors influencing pain severity. METHODS: This was a retrospective study included 340 women who underwent TV 4D-HyCoSy examination from January 2016 to October 2017. The factors were recorded, including age, childbearing history, infertility type, history of pelvic inflammation, pelvic surgery, history of uterine manipulation, history of ectopic pregnancy, atropine delivery mode, uterine position, uterine malformation, uterine lesion, fibroid, intrauterine adhesion, polycystic ovary, endometrial implantation cyst, dysmenorrhea score, the degree of patency of fallopian tube and contrast agents dosage. Pain was evaluated during and after TV 4D-HyCoSy. The time point of peak pain was evaluated and the influencing factors of moderate/severe pain were analyzed. RESULTS: The highest pain occurred at contrast instillation. The independent influencing factors of moderate/severe pain were age (P = 0.021), dysmenorrhea score (P = 0.003) and tubal patency (P < 0.001). Further analysis showed that age affected the pain score when TV 4D-HyCoSy started and the peak pain occurred. Dysmenorrhea score and tubal patency affect the pain score at most time points. CONCLUSION: Age, dysmenorrhea score and tubal patency are factors influencing the severity of pain during TV 4D-HyCoSy.

Effects of Different Degrees of Carotid Artery Stenosis on the Expression of XIAP and Smac in the Ischemic Penumbra of Rats with Cerebral Ischemia-Reperfusion.

OBJECTIVE: To investigate the effects of different degrees of carotid artery stenosis (CAS) on the expression of XIAP and Smac in ischemic penumbra of rats with cerebral ischemia-reperfusion (I/R). MATERIALS AND METHODS: Samples were collected at 12 h and 24 h after reperfusion, and then the treated groups were divided into the NC-12 group, NC-24 group, MIS-12 group, MIS-24 group, MOS-12 group, MOS-24 group, SES-12 group and SES-24 group. HE staining was used to observe the pathological changes of the brain tissue. TUNEL assay was used to detect the apoptosis in the ischemic penumbra. IHC and RT-qPCR were used to detect the expression of XIAP and Smac in the brain tissue. RESULTS: By observing the pathological sections of brain tissue, the rats in MIS, MOS and SES groups showed loose brain tissue on the infarcted side and neuronal pyknosis in the ischemic penumbra. And with the aggravation and prolongation of the degree of stenosis, the degree of brain injury deepened. It was further found that the TUNEL positive rate was significantly increased in the ischemic penumbra in the SES and MOS groups compared with that in the normal control (NC) group. The results of IHC and RT-qPCR showed that the mRNA expression of XIAP and Smac in the ischemic penumbra was significantly up-regulated in the MIS, MOS and SES groups compared with that in the NC group. CONCLUSIONS: CAS may activate XIAP/Smac signaling pathway to induce neuronal apoptosis and promote the injury in the ischemic penumbra caused by cerebral I/R.

Risk factors for ineffective recanalization after endovascular treatment in acute ischemic stroke.

OBJECTIVE: To assess the risk factors of early ineffective recanalization, defined as a modified Rankin scale (mRS) score of 4∼6 at two weeks after symptom onset, in acute ischemic stroke (AIS) patients receiving endovascular treatment (EVT). METHOD: One hundred eighty-four AIS patients who received EVT in 01/2016-12/2018 were analyzed. The recanalization rate was 81.5 % (150/184). Patients were divided into effective (mRS 0-3) and ineffective (mRS 4-6) recanalization groups. The independent risk factors for ineffective recanalization were analyzed. RESULT: The ineffective recanalization rate was 67.3 % (101/150 cases) at 14 days after symptom onset. Twenty-five patients (24.8 %) in the ineffective recanalization group had coronary heart disease, higher than that in the effective group (4 patients, 8.2 %, P = 0.016). Infratentorial pathology accounted for 34.7 % (35 cases) of the ineffective group and only 14.3 % (7 cases) of the control group (P = 0.011). Systolic blood pressure (SBP) was higher in the ineffective group than in the effective group (146 mmHg vs 140 mmHg, P = 0.038). General anesthesia was more common in the ineffective group than in the control group (49 cases, 48.5 %, vs 9 cases, 18.4 %; P = 0.000). Logistic regression showed that coronary heart disease (odds ratio (OR) = 3.399, 95 % confidence interval (CI) 1.013-11.401, P = 0.048), unconsciousness before EVT (OR = 4.292, 95 % CI 1.963-9.386, P = 0.000), SBP (OR = 1.016, 95 % CI 0.997-1.035, P = 0.090) and general anesthesia (OR = 3.378, 95 % CI 1.395-8.180, P = 0.007) were independent risk factors for ineffective recanalization. CONCLUSION: Patients with coronary heart disease, unconsciousness before EVT, elevated SBP and general anesthesia need precise assessment to benefit from EVT.

Palliative Radiofrequency Ablation Accelerates the Residual Tumor Progression Through Increasing Tumor-Infiltrating MDSCs and Reducing T-Cell-Mediated Anti-Tumor Immune Responses in Animal Model.

Previous studies showed that radiofrequency ablation (RFA) has a favorable treatment efficacy for hepatocellular carcinoma (HCC) or colorectal liver metastases (CRLMs). Palliative RFA (pRFA) resulting from larger HCC or multiple CRLMs further accelerated the progression of potential residual tumor, yet its mechanism was still unknown. This study investigated the influence of myeloid-derived suppressor cells (MDSCs) on T-cell immune responses and tumor recurrence after pRFA. CT26 tumor models were used. The percentage of MDSCs in peripheral blood was analyzed by flow cytometry after pRFA. The level of Th1 and Th2 cytokines were measured by ELISA through different treatments (n = 4/group). The tumor-infiltrating MDSCs, dendritic cells, and intracellular cytokines level were analyzed by IHC staining after different treatments. The functional CD8+ T cells were confirmed by the co-localization immunofluorescence staining. The long-term outcomes were also evaluated through CT26 and 4T1 tumor models. The results showed that tumor models treated with pRFA displayed significant increases in the percentage of MDSCs of peripheral blood and tumor infiltration. The expression level of TGF-ß and IL-6 after pRFA was higher than that before pRFA by ELISA and IHC staining. After depleting MDSCs by combining with Abs, the pRFA + Abs group achieved a higher level of Th1 cytokines and greatly enhanced the percentage of tumor-infiltrating functional CD8+ T cells when compared with pRFA alone. The depletion of MDSCs through combination with Abs also resulted in tumor regression. In conclusion, pRFA accelerates the residual tumor progression through increasing tumor-infiltrating MDSCs and reducing T-cell-mediated anti-tumor immune responses, which could provide a potential approach for delaying tumor recurrence caused by pRFA.

Spontaneous conception outcome in infertile women after four-dimensional hysterosalpingo-contrast-sonography.

BACKGROUND: Four-dimensional hysterosalpingo-contrast sonography (4D-HyCoSy) is the preferred way for evaluating fallopian tubal patency and it associated with higher rate of spontaneous conception. However, Few studies have evaluated the influencing factors of spontaneous conception in 4D-HyCoSy and suggested ways to choose treatment options after 4D-HyCoSy. The study was to evaluate the correlation between spontaneous conception outcome and the patients' clinical characteristics as well as tubal patency in infertile women to provide reference on ways to manage the patient after 4D-HyCoSy. METHODS: This was a retrospective study and analysis of two hundred and eighty three (283) infertile patients who underwent a 4D-HyCoSy between December 2014 and October 2017 in our center. Eligible patients were those whose partners semen parameters were normal when based on World Health Organization (WHO) criteria, and had spontaneous conception without clinical interventions after 4D-HyCoSy. RESULT(S): One hundred and sixteen patients (40.9%) conceived spontaneously and the mean conception time was (8.8 ± 0.3) months. Within a year after 4D-HyCoSy, the spontaneous conception rate was highest in type VI(62.5%), followed by type IV (46.2%), type III (44.4%), type V (39.4%), type II (33.9%) and type I (14.8%). With Cox regression analysis, two factors associated with spontaneous conception outcome appeared to increase spontaneous conception rate: patients with type IV or type VI tubes and duration of infertility less than 2 years. The age, type of infertility, multiparas, history of pelvic surgery, history of uterine cavity operation, uterine fibromyomata and polycystic ovary were unrelated to spontaneous conception outcome after 4D-HyCoSy. CONCLUSION(S): This study showed that some infertile women could succeed in spontaneous conception after 4D-HyCoSy. Hence, We recommend the usage of 4D-HyCoSy as first line for tubal patency test and infertile patients should be advised to accept 4D-HyCoSy examination as soon as possible. Expectant treatment of about 8-9 months is reported to be feasible for infertile women whose 4D-HyCoSy findings showed one tube patency or poor patency. Alternatively, an immediate clinical intervention is recommended for those with bilateral obstructed tubes .

[NF-κB inhibitor improves pulmonary vascular remodeling by reversing LPS-induced down-regulation of BMPRII].

The occurrence and development of pulmonary arterial hypertension (PAH) is closely related to the genetic mutation of bone morphogenetic protein receptor type II (BMPRII) encoding gene and the inflammatory response mediated by nuclear factor κB (NF-κB) pathway. This paper was aimed to investigate the effect of NF-κB pathway inhibitors on lipopolysaccharide (LPS)-induced pulmonary artery endothelial cell injury. Human pulmonary artery endothelial cells were treated with 1 µg/mL of LPS. The expression levels of BMPRII and interleukin-8 (IL-8) were detected by Western blot and qPCR. The rat PAH model was established by intraperitoneal (i.p.) injection of monocrotaline (MCT). The expression levels of BMPRII and IL-8 in pulmonary artery endothelial cells were detected by immunofluorescence staining. Cardiac hemodynamic changes and pulmonary vascular remodeling were detected in the MCT-PAH model rats. The results showed that LPS caused down-regulation of BMPRII expression and up-regulation of IL-8 expression in human pulmonary artery endothelial cells. NF-κB inhibitor BAY11-7082 (10 µmol/L) reversed the effect of LPS. In the pulmonary artery endothelial cells of MCT-PAH model, BMPRII expression was down-regulated, IL-8 expression was up-regulated, weight ratio of right ventricle to left ventricle plus septum [RV/(LV+S)] and right ventricular systolic pressure (RVSP) were significantly increased, cardiac output (CO) and tricuspid annular plane systolic excursion (TAPSE) were significantly reduced, and pulmonary vessel wall was significantly thickened. BAY11-7082 (5 mg/kg, i.p., 21 consecutive days) reversed the above changes in the MCT-PAH model rats. These results suggest that LPS down-regulates the expression level of BMPRII through NF-κB signaling pathway, promoting the occurrence and development of PAH. Therefore, the NF-κB pathway can be used as a potential therapeutic target for PAH.

Near-infrared fluorescence imaging-guided focused ultrasound-mediated therapy against Rheumatoid Arthritis by MTX-ICG-loaded iRGD-modified echogenic liposomes.

Rheumatoid arthritis (RA), a common inflammatory disorder of the joints characterized by synovitis and pannus formation, often results in irreversible joint erosion and disability. Methotrexate (MTX) is the first-line drug against RA, but the therapeutic effects are sub-optimal due to its poor retention at the target sites and systemic side effects. Multifunctional nanoparticles are highly promising agents for minimally invasive, traceable and effective targeted therapy. Methods: This study developed iRGD peptide-functionalized echogenic liposomes (iELPs) which encapsulates MTX and indocyanine green (ICG) fluorescent probe through the thin film-hydration method. Results: The resulting iELPs showed high affinity for endothelial cells overexpressing αvß3 integrin, favorable acoustic response and fluorescence tracking properties. Also, near-infrared (NIR) fluorescence imaging of iELPs and ultrasound-triggered drug release of MTX were proved in a mouse RA model, greatly improving the therapeutic efficacy and reducing MTX side effects. Histological assessment of the articular tissues further revealed significantly lower inflammatory cell infiltration and angiogenesis in the iELPs-treated and sonicated mice. Conclusion: Our study provides a promising nanoplatform for integrating ultrasound-controlled drug release and NIR fluorescence imaging for RA treatment.

Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development.

Vascular development is determined by the sequential expression of specific genes, which can be studied by performing in situ hybridization assays in zebrafish during different developmental stages. To investigate the role of endoglin(eng) in vessel formation during the development of hereditary hemorrhagic telangiectasia (HHT), morpholino-mediated targeted knockdown of eng in zebrafish are used to study its temporal expression and associated functions. Here, whole-mount in situ RNA hybridization (WISH) is employed for the analysis of eng and its downstream genes in zebrafish embryos. Also, tube formation assays are performed in HHT patient-derived induced pluripotent stem cell-differentiated endothelial cells (iPSC-ECs; with eng mutations). A specific signal amplifying system using the whole amount In Situ Hybridization - WISH provides higher resolution and lower background results compared to traditional methods. To obtain a better signal, the post-fixation time is adjusted to 30 min after probe hybridization. Because fluorescence staining is not sensitive in zebrafish embryos, it is replaced with diaminobezidine (DAB) staining here. In this protocol, HHT patient-derived iPSC lines containing an eng mutation are differentiated into endothelial cells. After coating a plate with basement membrane matrix for 30 min at 37 °C, iPSC-ECs are seeded as a monolayer into wells and kept at 37 °C for 3 h. Then, the tube length and number of branches are calculated using microscopic images. Thus, with this improved WISH protocol, it is shown that reduced eng expression affects endothelial progenitor formation in zebrafish embryos. This is further confirmed by tube formation assays using iPSC-ECs derived from a patient with HHT. These assays confirm the role for eng in early vascular development.

Evidence of Accumulated Endothelial Progenitor Cells in the Lungs of Rats with Pulmonary Arterial Hypertension by 89Zr-oxine PET Imaging.

Endothelial progenitor cells (EPCs) play a major role in regulating pulmonary vascular remodeling during pulmonary arterial hypertension (PAH) development. Several preclinical and clinical trials of EPCs transplantation have been performed for the treatment of PAH. However, there is no reliable method to monitor real-time cell trafficking and quantify transplanted EPCs. Here in this paper we isolated EPCs from human peripheral blood, identified their functional integrity, and efficiently labeled the EPCs with 89Zr-oxine and DiO. Labeled EPCs were injected into the tail vein of normal and PAH rats to be tracked in vivo. From the microPET/CT images, we found EPCs were distributed primarily in the lung at 1 h and then migrated to the liver and spleen. We could observe the 3,3' dioctadecyloxacarbocyanine perchlorate (DiO)-labeled EPCs binding in the pulmonary vasculature by CellVizio confocal. The result of quantitative analysis revealed significantly higher accumulation of EPCs in the lungs of PAH rats than in those of healthy rats. The distribution and higher accumulation of EPCs in the lungs of PAH rats could help to evaluate the safety and provide evidence of effectiveness of EPC therapy.

Theranostic nanosensitizers for highly efficient MR/fluorescence imaging-guided sonodynamic therapy of gliomas.

Glioma is the most frequent primary brain tumour of the central nervous system. Its high aggressiveness and deep-seated brain lesion make it a great challenge to develop a non-invasive, precise and effective treatment approach. Here, we report a multifunctional theranostic agent that can integrate imaging and therapy into a single nano-platform for imaging-guided sonodynamic therapy (SDT). The SDT agents were fabricated by encapsulation of sinoporphyrin sodium (DVDMS) chelating with manganese ions into nanoliposomes (DVDMS-Mn-LPs). DVDMS-Mn-LPs are physiologically stable and biologically compatible, and they can produce singlet oxygen upon ultrasound irradiation to kill cancer cells. Both cell and animal studies demonstrated that SDT with DVDMS-Mn-LPs can significantly improve the antitumour growth efficiency even in the presence of skull. In addition, DVDMS-Mn-LPs are good for MR and fluorescence imaging. Thus, DVDMS-Mn-LPs reported here may provide a promising strategy for imaging-guided modality for glioma treatment.

Fast preparation of carbon spheres from enzymatic hydrolysis lignin: Effects of hydrothermal carbonization conditions.

This study explores the effect of carbon sphere preparation conditions on the morphology of the carbon spheres and the micropore development by fast potassium hydroxide activation via microwave heating. Enzymatic hydrolysis lignin is used as the precursor for carbon sphere preparation via environmentally friendly hydrothermal carbonization. The effects of various carbonization temperatures, carbonization times and reaction concentrations on the physical morphology of the carbon sphere surfaces are investigated. The Brunauer-Emmett-Teller surface area, yield and scanning electron microscopic images are used to characterize the carbon spheres. High carbonization temperatures and times result in large particle sizes, high sphericity, uniform size, and high dispersity of the carbon spheres. The best carbon spheres are obtained at 270 °C for 7 hours with a reaction concentration of 0.06 g ml-1 and a particle size of 3-6 µm. After activation, the Brunauer-Emmett-Teller surface area of the activated carbon spheres increases from 248 m2 g-1 to 1278 m2 g-1. Carbon spheres activated by treatment with fast potassium hydroxide and microwave heating can develop micropores that enhance the adsorptive capacity for small molecules, such as gases. Enzymatic hydrolysis lignin-derived carbon spheres formed via hydrothermal carbonization should be potentially sustainable materials applicable in energy and environmental fields.

Synovitis in mice with inflammatory arthritis monitored with quantitative analysis of dynamic contrast-enhanced NIR fluorescence imaging using iRGD-targeted liposomes as fluorescence probes.

BACKGROUND: Rheumatoid arthritis (RA) is a common inflammatory disorder characterized primarily by synovitis and pannus formation in multiple joints, causing joints destruction and irreversible disability in most cases. Early diagnosis and effective therapy monitoring of RA are of importance for achieving the favorable prognosis. METHODS: We first prepared the targeted fluorescence probes, and then explored the feasibility of near-infrared (NIR) fluorescence molecular imaging to detect and evaluate the RA via the targeted fluorescence probes by quantitative analysis in this study. RESULTS: The targeted fluorescence probes (indocyanine green-liposomes decorated with iRGD peptide [iLPs]) was successfully prepared. The quantitative analysis found that strong fluorescence signal was detected in inflamed paws and the fluorescence signal in iLPs group was 3.03-fold higher than that in non-targeted (indocyanine green-liposomes decorated without iRGD peptide [LPs]) group (P<0.01) at 15 min after injection, whereas the fluorescence signal from iLPs signal can almost not be observed in the non-inflamed paws, showing the high sensitivity and accuracy for arthritis by the NIR fluorescence imaging based on iLPs. CONCLUSION: The NIR fluorescence imaging by iLPs may facilitate improved arthritis diagnosis and early assessment of the disease progression by providing an in vivo characterization of angiogenesis in inflammatory joint diseases.