Ferrous Ion Alleviates Lipid Deposition and Inflammatory Responses Caused by a High Cottonseed Meal Diet by Modulating Hepatic Iron Transport Homeostasis and Controlling Ferroptosis in Juvenile Ctenopharyngodon idellus.

To investigate the mechanisms through which ferrous ion (Fe2+) addition improves the utilization of a cottonseed meal (CSM) diet, two experimental diets with equal nitrogen and energy content (low-cottonseed meal (LCM) and high-cottonseed meal (HCM) diets, respectively) containing 16.31% and 38.46% CSM were prepared. Additionally, the HCM diet was supplemented with graded levels of FeSO4·7H2O to establish two different Fe2+ supplementation groups (HCM + 0.2%Fe2+ and HCM + 0.4%Fe2+). Juvenile Ctenopharyngodon idellus (grass carps) (5.0 ± 0.5 g) were fed one of these four diets (HCM, LCM, HCM + 0.2%Fe2+ and HCM + 0.4%Fe2+ diets) for eight weeks. Our findings revealed that the HCM diet significantly increased lipid peroxide (LPO) concentration and the expression of lipogenic genes, e.g., sterol regulatory element binding transcription factor 1 (srebp1) and stearoyl-CoA desaturase (scd), leading to excessive lipid droplet deposition in the liver (p < 0.05). However, these effects were significantly reduced in the HCM + 0.2%Fe2+ and HCM + 0.4%Fe2+ groups (p < 0.05). Plasma high-density lipoprotein (HDL) concentration was also significantly lower in the HCM and HCM + 0.2%Fe2+ groups compared to the LCM group (p < 0.05), whereas low-density lipoprotein (LDL) concentration was significantly higher in the HCM + 0.2%Fe2+ and HCM + 0.4%Fe2+ groups than in the LCM group (p < 0.05). Furthermore, the plasma levels of liver functional indices, including alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and glucose (GLU), were significantly lower in the HCM + 0.4%Fe2+ group (p < 0.05). Regarding the expression of genes related to iron transport regulation, transferrin 2 (tfr2) expression in the HCM group and Fe2+ supplementation groups were significantly suppressed compared to the LCM group (p < 0.05). The addition of 0.4% Fe2+ in the HCM diet activated hepcidin expression and suppressed ferroportin-1 (fpn1) expression (p < 0.05). Compared to the LCM group, the expression of genes associated with ferroptosis and inflammation, including acyl-CoA synthetase long-chain family member 4b (acsl4b), lysophosphatidylcholine acyltransferase 3 (lpcat3), cyclooxygenase (cox), interleukin 1ß (il-1ß), and nuclear factor kappa b (nfκb), were significantly increased in the HCM group (p < 0.05), whereas Fe2+ supplementation in the HCM diet significantly inhibited their expression (p < 0.05) and significantly suppressed lipoxygenase (lox) expression (p < 0.05). Compared with the HCM group without Fe2+ supplementation, Fe2+ supplementation in the HCM diet significantly upregulated the expression of genes associated with ferroptosis, such as heat shock protein beta-associated protein1 (hspbap1), glutamate cysteine ligase (gcl), and glutathione peroxidase 4a (gpx4a) (p < 0.05), and significantly decreased the expression of the inflammation-related genes interleukin 15/10 (il-15/il-10) (p < 0.05). In conclusion, FeSO4·7H2O supplementation in the HCM diet maintained iron transport and homeostasis in the liver of juvenile grass carps, thus reducing the occurrence of ferroptosis and alleviating hepatic lipid deposition and inflammatory responses caused by high dietary CSM contents.

TNF-α and IL-8 levels are positively correlated with hypobaric hypoxic pulmonary hypertension and pulmonary vascular remodeling in rats.

The expression status of proinflammatory cytokines in high-altitude pulmonary arterial hypertension (PAH) has been well studied. However, the changes in interleukin (IL)-8 and tumor necrosis factor α (TNF-α) during the reversible changes in pulmonary vascular remodeling (PVR) in PAH after detaching from a hypobaric hypoxic environment have not been elucidated. This investigation elucidated a high-altitude PAH rat model. Then, PAH rats in the high-altitude group were maintained in the high-altitude area, and rats in the low-altitude group returned to the low-altitude area. After 0, 10, 20, and 30 days of PAH modeling, right ventricular systolic pressure (RVSP) and the mean pulmonary arterial pressure (mPAP) were assessed. Right ventricular (RV) hypertrophy was reflected by the ratio of RV/[left ventricle + interventricular septum (S)]. Pathological changes in PVR were accessed by hematoxylin-eosin staining, and medial wall thickness (WT%) and medial wall area (WA%) were measured. TNF-α and IL-8 levels in pulmonary artery tissues and blood were measured with Western blot assay and enzyme-linked immunosorbent assay, respectively. Our results showed that PAH rats exhibited a substantial increase in RVSP and mPAP, RV hypertrophy, PVR, and enhanced generation of TNF-α and IL-8. Then, we found that these pathological changes were gradually aggravated and TNF-α and IL-8 levels were increased in rats in the high-altitude group after 10, 20, and 30 days of PAH modeling. In contrast, the mPAP was decreased and PVR was alleviated in rats in the low-altitude group, accompanying with reduced TNF-α and IL-8 production. In conclusion, our study demonstrated that the generation of TNF-α and IL-8 was also reversible during the reversible changes in PVR after detaching from a hypobaric hypoxic environment. Thus, proinflammatory cytokine TNF-α and IL-8 levels are positively correlated with PVR severity.

Predicting Extrathyroidal Extension in Papillary Thyroid Carcinoma Using a Clinical-Radiomics Nomogram Based on B-Mode and Contrast-Enhanced Ultrasound.

Papillary thyroid carcinoma (PTC) is the most common pathological type of thyroid cancer. PTC patients with extrathyroidal extension (ETE) are associated with poor prognoses. The preoperative accurate prediction of ETE is crucial for helping the surgeon decide on the surgical plan. This study aimed to establish a novel clinical-radiomics nomogram based on B-mode ultrasound (BMUS) and contrast-enhanced ultrasound (CEUS) for the prediction of ETE in PTC. A total of 216 patients with PTC between January 2018 and June 2020 were collected and divided into the training set (n = 152) and the validation set (n = 64). The least absolute shrinkage and selection operator (LASSO) algorithm was applied for radiomics feature selection. Univariate analysis was performed to find clinical risk factors for predicting ETE. The BMUS Radscore, CEUS Radscore, clinical model, and clinical-radiomics model were established using multivariate backward stepwise logistic regression (LR) based on BMUS radiomics features, CEUS radiomics features, clinical risk factors, and the combination of those features, respectively. The diagnostic efficacy of the models was assessed using receiver operating characteristic (ROC) curves and the DeLong test. The model with the best performance was then selected to develop a nomogram. The results show that the clinical-radiomics model, which is constructed by age, CEUS-reported ETE, BMUS Radscore, and CEUS Radscore, showed the best diagnostic efficiency in both the training set (AUC = 0.843) and validation set (AUC = 0.792). Moreover, a clinical-radiomics nomogram was established for easier clinical practices. The Hosmer-Lemeshow test and the calibration curves demonstrated satisfactory calibration. The decision curve analysis (DCA) showed that the clinical-radiomics nomogram had substantial clinical benefits. The clinical-radiomics nomogram constructed from the dual-modal ultrasound can be exploited as a promising tool for the pre-operative prediction of ETE in PTC.

Ultrasonography-based radiomics and computer-aided diagnosis in thyroid nodule management: performance comparison and clinical strategy optimization.

Objectives: To compare ultrasonography (US) feature-based radiomics and computer-aided diagnosis (CAD) models for predicting malignancy in thyroid nodules, and to evaluate their utility for thyroid nodule management. Methods: This prospective study included 262 thyroid nodules obtained between January 2022 and June 2022. All nodules previously underwent standardized US image acquisition, and the nature of the nodules was confirmed by the pathological results. The CAD model exploited two vertical US images of the thyroid nodule to differentiate the lesions. The least absolute shrinkage and operator algorithm (LASSO) was applied to choose radiomics features with excellent predictive properties for building a radiomics model. Ultimately, the area under the receiver operating characteristic curve (AUC) and calibration curves were assessed to compare diagnostic performance between the models. DeLong's test was used to analyze the difference between groups. Both models were used to revise the American College of Radiology Thyroid Imaging Reporting and Data Systems (ACR TI-RADS) to provide biopsy recommendations, and their performance was compared with the original recommendations. Results: Of the 262 thyroid nodules, 157 were malignant, and the remaining 105 were benign. The diagnostic performance of radiomics, CAD, and ACR TI-RADS models had an AUC of 0.915 (95% confidence interval (CI): 0.881-0.947), 0.814 (95% CI: 0.766-0.863), and 0.849 (95% CI: 0.804-0.894), respectively. DeLong's test showed a statistically significant between the AUC values of models (p < 0.05). Calibration curves showed good agreement in each model. When both models were applied to revise the ACR TI-RADS, our recommendations significantly improved the performance. The revised recommendations based on radiomics and CAD showed an increased sensitivity, accuracy, positive predictive value, and negative predictive value, and decreased unnecessary fine-needle aspiration rates. Furthermore, the radiomics model's improvement scale was more pronounced (33.3-16.7% vs. 33.3-9.7%). Conclusion: The radiomics strategy and CAD system showed good diagnostic performance for discriminating thyroid nodules and could be used to optimize the ACR TI-RADS recommendation, which successfully reduces unnecessary biopsies, especially in the radiomics model.

Clinical-Radiomics Nomogram Based on Contrast-Enhanced Ultrasound for Preoperative Prediction of Cervical Lymph Node Metastasis in Papillary Thyroid Carcinoma.

This study aimed to establish a new clinical-radiomics nomogram based on ultrasound (US) for cervical lymph node metastasis (LNM) in papillary thyroid carcinoma (PTC). We collected 211 patients with PTC between June 2018 and April 2020, then we randomly divided these patients into the training set (n = 148) and the validation set (n = 63). 837 radiomics features were extracted from B-mode ultrasound (BMUS) images and contrast-enhanced ultrasound (CEUS) images. The maximum relevance minimum redundancy (mRMR) algorithm, least absolute shrinkage and selection operator (LASSO) algorithm, and backward stepwise logistic regression (LR) were applied to select key features and establish a radiomics score (Radscore), including BMUS Radscore and CEUS Radscore. The clinical model and clinical-radiomics model were established using the univariate analysis and multivariate backward stepwise LR. The clinical-radiomics model was finally presented as a clinical-radiomics nomogram, the performance of which was evaluated by the receiver operating characteristic curves, Hosmer-Lemeshow test, calibration curves, and decision curve analysis (DCA). The results show that the clinical-radiomics nomogram was constructed by four predictors, including gender, age, US-reported LNM, and CEUS Radscore. The clinical-radiomics nomogram performed well in both the training set (AUC = 0.820) and the validation set (AUC = 0.814). The Hosmer-Lemeshow test and the calibration curves demonstrated good calibration. The DCA showed that the clinical-radiomics nomogram had satisfactory clinical utility. The clinical-radiomics nomogram constructed by CEUS Radscore and key clinical features can be used as an effective tool for individualized prediction of cervical LNM in PTC.

An indolizine squaraine-based water-soluble NIR dye for fluorescence imaging of multidrug-resistant bacteria and antibacterial/antibiofilm activity using the photothermal effect.

The majority of nosocomial infections are caused by bacteria with antimicrobial resistance and the formation of biofilms, such as implant-related bacterial infections and sepsis. There is an urgent need to develop new strategies for early-stage screening, destruction of multidrug-resistant bacteria, and efficient inhibition of biofilms. Organic dyes that absorb and emit in the near-infrared (NIR) region are potentially non-invasive, high-resolution, and rapid biological imaging materials. In this study, a non-toxic and biocompatible indolizine squaraine dye with water-solubilizing sulfonate groups (SO3SQ) is studied for bacterial imaging and photothermal therapy (PTT). PTT is efficient in eliminating microorganisms through local hyperthermia without the risk of developing drug-resistant bacteria. The optical properties of SO3SQ are studied extensively in phosphate-buffered saline (PBS). UV-Vis-NIR absorption spectra analysis shows a strong absorption between 650 nm - 1000 nm. SO3SQ allows for the wash-free fluorescence imaging of drug-resistant bacteria via NIR fluorescence imaging due to a "turn-on" fluorescence property of the dye when interacting with bacteria. Although SO3SQ exhibits no toxicity against both Gram-positive bacteria and Gram-negative bacteria, the PTT property of SO3SQ is efficient in killing bacteria as well as inhibiting and eradicating biofilms. PTT experiments demonstrate that SO3SQ reduces 90% of cell viability in bacterial strains under NIR radiation with a minimum inhibition concentration (MIC90) of >450 µg/mL. The PTT property of SO3SQ can also inhibit biofilms (BIC90 = 1000-2000 µg/mL) and eradicate both preformed young and mature biofilms (MBEC90 = 1500-2000 µg/mL) as observed by crystal violet assays.

Comparison of clinical characteristics, treatment efficacy and survival in patients with newly diagnosed multiple myeloma with single- versus multi-site extramedullary invasion.

PURPOSE: We aimed to compare the clinical characteristics, treatment efficacy and survival in patients with newly diagnosed multiple myeloma with single- versus multi-site extramedullary invasion. METHODS: A total of 90 extramedullary multiple myeloma (EMM) patients were included. We compared the characteristics, overall remission rates (ORRs) and survival among patients with single-site and multi-site invasions. Comparison was also done regarding extramedullary-bone-related (EM-B) and extramedullary-extraosseous (EM-E) multiple myeloma patients. RESULTS: Patients with multi-site invasion had higher LDH (179.0 U/L) than single-site invasion (P = 0.016). Significantly higher LDH was also showed in patients with EM-E (189.4 U/L) than EM-B (P = 0.025). The ORR of patients with single-site invasion (72.1%) was not significantly higher than multi-site invasion (68.2%) (P = 0.690). But the ORR of patients with EM-B was significantly higher than EM-E (78.2 vs. 56.3%, P = 0.031). Among patients with single-site invasion, the multivariate survival analysis showed that PI plus IMiD regimen significantly improved the PFS (P < 0.05). Among patients with multi-site invasion or with EM-B, the multivariate survival analysis confirmed the associations of rISS III with poor PFS and OS (P < 0.05). Among patients with EM-E, plasma cell percentage ≥ 20% was associated with significantly poor PFS and OS (P < 0.05). CONCLUSION: rISS stage III was possibly associated with poor survival of EMM patients with multi-site invasion or with EM-B. Plasma cell percentage ≥ 20% was associated with poor survival of EMM patients with EM-E. Comparison should not only be done between single- and multi-site invasions, but also between EM-B and EM-E.

A 5-year follow-up study on the efficacy and safety of ultrasound-guided laser ablation in elderly patients with papillary thyroid microcarcinoma: A retrospective, single-center study from China.

Objective: The aim of this study is to evaluate the long-term efficacy and safety of ultrasound-guided percutaneous laser ablation (PLA) for the treatment of elderly patients with papillary thyroid microcarcinoma (PTMC). Methods: From September 2015 to April 2017, 38 elderly patients with PTMC confirmed through fine-needle aspiration biopsy (FNAB) were treated with PLA. Before the treatment, the location and volume of the nodule together with the patients' symptoms were evaluated. Twenty-four hours after the treatment, contrast-enhanced ultrasound (CEUS) was performed to evaluate the completeness of the ablation. To evaluate the volume of the ablation area and recurrence or metastasis, ultrasound examination was performed at 1, 3, 6, and 12 months after the treatment and every 6 months thereafter. FNAB was performed for any suspicious recurrence or metastasis lesions. Result: The ablation of all the 38 patients was all achieved completely as confirmed by CEUS. No obvious complications were found. The success rate of single ablation was 100%. The average follow-up time was 64.58 ± 5.29 months (60-78 months). By the time of the last follow-up, 31 (81.58%) ablation lesions disappeared completely and seven (18.42%) ablation lesions showed scar-like changes. The volume of nodules was 40.69 ± 16.45 mm3 before operation, which decreased to 0.22 ± 0.76 mm3 by the end of 42 months, and all nodules disappeared 4 years after ablation (P < 0.01). At 6, 12, 18, 24, 30, 36, and 42 months after ablation, the average volume reduction rates (VRRs) were 12.09%, 31.21%, 50.9%, 72.06%, 84.79%, 95.65%, and 100%, respectively. Of all the patients enrolled, one patient (2.6%) had local recurrence and was treated with PLA again. No regrowth of treated nodule or lymph node metastasis and distant metastases was detected. Conclusion: Ultrasound-guided PLA is effective and safe for the treatment of elderly patients with PTMC who are ineligible for surgery.

The value of ultrasound guided laser ablation in papillary thyroid recurrence carcinoma: A retrospective, single center study from China.

Objective: The efficacy and safety of ultrasound-guided percutaneous laser ablation (PLA) for treating recurrent papillary thyroid cancer nodules (RPTCNs). Methods: A retrospective study was conducted in 43 patients with single recurrent thyroid cancer which was diagnosed by fine needle aspiration biopsy (FNAB). The extent of ablation was assessed by contrast-enhanced ultrasound (CEUS) 24h after PLA. At baseline (before ablation), 6, and 12 months, and every 6 months thereafter, the following were recorded: nodule maximum diameter, volume reduction rate (VRR), complications, and side effects. Result: All 43 patients were successfully treated with PLA without serious complications. All patients underwent CEUS 24 hours after PLA treatment, and all achieved complete ablation. The success rate of single ablation was 100%. The average follow-up time was 23.47 ± 6.50 months, 12 ~ 36 months. At the last follow-up, 32 (74.4%) ablation lesions disappeared completely and 11 (25.6%) ablation lesions showed scar-like changes. No lymph node metastasis was found during follow-up. The maximum diameter and volume of nodules decreased from 5.1 ± 1.4 mm, 86.22 ± 20.46 mm3 before operation to 0.73 ± 1.1 mm, 1.02 ± 1.92 mm3 at the end of observation (P < 0.01). The average volume reduction rates (VRR) at 6, 12, 18, 24, 30 and 36 months after ablation were 11.92%, 60.64%, 82.26%, 90.96%, 93.7% and 97.79% respectively. No regrowth of treated nodule and distant metastases were detected. One patient (2.3%) had local recurrence and was treated with PLA again. Conclusion: Ultrasound-guided PLA appears to be effective and safe for treating unifocal RPTCNs in selected patients who are ineligible for surgery, which is suitable for clinical application and promotion.

Summary of diagnostic and characteristic parameters of C0 to C2 patients based on the CEAP2020 classification.

OBJECTIVES: Hemodynamic changes in C0 to C2 according to the clinical/etiological/anatomical/pathophysiological classification have not been analyzed extensively. We intend to investigate the characteristics of early stage venous disease using venous clinical severity score (VCSS), heaviness/ache/swelling/throbbing/itching (HASTI) score and duplex ultrasound (DUS) derived parameters. METHODS: From Aug. 2020 to Jul. 2021, consecutive patients were categorized according to the clinical/etiological/anatomical/pathophysiological 2020 classification. The vein diameter (mm), reflux time (s), cross-sectional area (cm2), peak and mean reflux velocity (cm/s), and total reflux volume (mL) were documented in the superficial system and deep system. VCSS and HASTI scores were assessed and DUS parameters were analyzed. A P value of less than .05 was considered statistically significant. RESULTS: We studied 257 consecutive patients (142 female) with 371 limbs. The mean age was 50.96 ± 13.27 years (range, 20-81 years; median, 53 years) with an average body mass index of 24.03 ± 2.96 kg/m2 (range, 17.00-33.06 kg/m2; median, 23.67 kg/m2). The proportion of clinical (C) category was as followed: 47 C0S (12.67%), 45 C1 (12.13%), and 279 C2 (75.20%). There were 42.6% of C0 and 62.2% of C1 associated with some form of venous reflux, Pr (pathology [P]-reflux). Reflux of the great saphenous vein above the knee (GSVa) was the most commonly affected vein segment from C0-2. C1 differed from C0 only by age and severity scores with a cut-off value of 1 for both VCSS and HASTI. Larger saphenous veins diameter (GSV and small saphenous vein) were seen in Pr (P-reflux) than Pn (P-none) of C0 cases. Larger deep vein diameters (common femoral vein, femoral vein, and popliteal vein) were evident in Pr versus Pn of C1 cases. In C2, the popliteal vein, saphenofemoral junction, and GSVa were dilated in Pr cases. CONCLUSIONS: Both HASTI and VCSS were discriminative from C0 to C2. C0 differed from C1 by age and severity scores as DUS characteristics were not significantly different between the two groups. Vessel diameter played a discriminative role in distinguishing Pn versus Pr cases in each C0-2 category. GSVa was the most frequent reflux segment irrespective of reflux types.

A novel tissue-mimicking phantom for US/CT/MR-guided tumor puncture and thermal ablation.

AIM: This study aimed to develop a novel tumor-bearing tissue phantom model that can be used for US/CT/MR-guided tumor puncture and thermal ablation. METHODS: The phantom model comprised two parts: a normal tissue-mimicking phantom and a tumor-mimicking phantom. A normal tissue phantom was prepared based on a polyacrylamide gel mixed with thermochromic ink. Moreover, a spherical phantom containing contrast agents was constructed and embedded in the tissue phantom to mimic a tumor lesion. US/CT/MR imaging features and thermochromic property of the phantom model were characterized. Finally, the utility of the phantom model for imaging-guided microwave ablation training was examined. RESULTS: The tumor phantom containing contrast agents showed hyper-echogenicity, higher CT numbers, and lower T2 signal intensity compared with the normal tissue phantom in US/CT/MR images. Consequently, we could locate the position of the tumor in US/CT/MR imaging and perform an imaging-guided tumor puncture. When the temperature reached the threshold of 60 °C, the phantom exhibited a permanent color change from cream white to magenta. Based on this obvious color change, our phantom model could clearly map the thermal ablation region after thermotherapy. CONCLUSIONS: We developed a novel US/CT/MR-imageable tumor-bearing tissue model that can be used for imaging-guided tumor puncture and thermal ablation. Furthermore, it allows visual assessment of the ablation region by analyzing the obvious color change. Overall, this phantom model could be a good training tool in the field of thermal ablation.

Polyethylene glycol-coated ultrasmall superparamagnetic iron oxide nanoparticles-coupled sialyl Lewis X nanotheranostic platform for nasopharyngeal carcinoma imaging and photothermal therapy.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a type of head and neck malignant tumor with a high incidence in specific regional distribution, and its traditional therapies face some challenges. It has become an urgent need to seek new therapeutic strategies without or with low toxicity and side effects. At present, more and more researchers has been attracting attention by nanotheranostic platform. Therefore, our team synthesized the polyethylene glycol-coated ultrasmall superparamagnetic iron oxide nanoparticles-coupled sialyl Lewis X (USPIO-PEG-sLex) nanotheranostic platform with high temperature pyrolysis. RESULTS: The USPIO-PEG-sLex nanoparticles had excellent photothermal conversion property, and the temperature of USPIO-PEG-sLex nanoparticles solution increased with its concentration and power density of near-infrared (NIR) on 808 nm wavelengths. Five USPIO-PEG-sLex nanoparticles with different concentrations of 0 mg/ml, 0.025 mg/ml, 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml were prepared. The biological toxicity results showed that the viability of NPC 5-8F cells is related to the concentration of USPIO-PEG-sLex nanoparticles and the culture time (P < 0.001). The results of photothermal therapy (PTT) in vitro indicated that the viability of 5-8F cells decreased significantly with the concentration of USPIO-PEG-sLex nanoparticles increases (P < 0.001), and the viability of NPC 5-8F cells were 91.04% ± 5.20%, 77.83% ± 3.01%, 73.48% ± 5.55%, 59.50% ± 10.98%, 17.11% ± 3.14%, respectively. The USPIO-PEG-sLex nanoparticles could target the tumor area, and reduce the T2* value of tumor tissue. The T2* values of tumor pre- and post-injection were 30.870 ± 5.604 and 18.335 ± 4.351, respectively (P < 0.001). In addition, USPIO-PEG-sLex nanoparticles as a photothermal agent for PTT could effectively inhibit tumor progression. The ratio of volume change between tail vein injection group, control group, nanoparticles without laser irradiation group and blank group after 5 treatments were 3.04 ± 0.57, 5.80 ± 1.06, 8.09 ± 1.96, 7.89 ± 2.20, respectively (P < 0.001). CONCLUSIONS: Our synthesized USPIO-PEG-sLex nanotheranostic platform, and it may be become a new strategy for the treatment of NPC.

Versatile Surface Functionalization of Water-Dispersible Iron Oxide Nanoparticles with Precisely Controlled Sizes.

The synthesis of highly water-dispersible iron oxide nanoparticles with surface functional groups and precisely controlled sizes is essential for biomedical application. In this paper, we report a one-pot strategy for versatile surface functionalization. The iron oxide nanoparticles are first synthesized by thermal decomposition of iron(III) acetylacetonate (Fe(acac)3) in diethylene glycol (DEG), and their surfaces are modified by adding the surface ligands at the end of the reaction. The size of iron oxide nanoparticles can be precisely controlled in nanometer scale by continuous growth. This facile synthesis method enables the surface modification with different coating materials such as dopamine (DOPA), polyethylene glycol with thiol end group (thiol-PEG), and poly(acrylic acid) (PAA) onto the iron oxide nanoparticles, introducing new surface functionalities for future biomedical application. From transmission electron microscopy (TEM) and X-ray diffraction (XRD), the morphology and crystal structure are not changed during surface functionalization. The attachment of surface ligands is studied by Fourier transform infrared spectroscopy (FTIR) and Thermogravimetric Analysis (TGA). The surface functional groups are confirmed by X-ray Photoelectron Spectroscopy (XPS). In correlation with the change of hydrodynamic size, PAA coated nanoparticles are found to exhibit outstanding stability in aqueous solution. Furthermore, we demonstrate that the functional groups are available for conjugating with other molecules such as fluorescent dye, showing potential biological applications. Lastly, the magnetic resonance phantom studies demonstrate that iron oxide nanoparticles with PAA coating can be used as T1 and T2 dual-modality contrast agents. Both r1 and r2 relaxivities significantly increase after surface functionalization with PAA, indicating improved sensitivity.

Measurement for the Area of Red Blood Cells From Microscopic Images Based on Image Processing Technology and Its Applications in Aplastic Anemia, Megaloblastic Anemia, and Myelodysplastic Syndrome.

BACKGROUND: Aplastic anemia (AA), megaloblastic anemia (MA), and myelodysplastic syndrome (MDS) were common anemic diseases. Sometimes it was difficult to distinguish patients with these diseases. METHODS: In this article, we proposed one measurement method for the area of red blood cells (RBCs) from microscopic images based on image processing technology and analyzed the differences of the area in 25 patients with AA, 64 patients with MA, and 68 patients with MDS. RESULTS: The area of RBCs was 44.19 ± 3.88, 42.09 ± 5.35, 52.87 ± 7.68, and 45.75 ± 8.07 µm2 in normal subjects, patients with AA, MA, and MDS, respectively. The coefficients of variation were 8.78%, 10.05%, 14.53%, and 14.00%, respectively, in these groups. The area of RBCs in patients with MA was significantly higher than normal subjects (p < 0.001). Compared with patients with AA and MDS, the area of RBCs in patients with MA was also significantly higher (p < 0.001). The results of correlation analysis between the area of RBCs and mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), MCH concentration (MCHC), and red cell distribution width showed no significant correlations (p > 0.05). The area under the curve (AUC) results of the Receiver Operating Characteristic (ROC) curves of RBCs area were 0.421, 0.580, and 0.850, respectively, in patients with AA (p = 0.337), MDS (p = 0.237), and MA (p < 0.001). CONCLUSION: Identifying the area of RBCs in peripheral blood smears based on the image processing technology could achieve rapid and efficient diagnostic support for patients with MDS and MA, especially for patients with MA and in combination with MCV. However, a larger sample study is needed to find the cutoff area values.

Association between vein diameters, reflux characteristics, and clinical severity in patients with chronic venous insufficiency in Northwest China.

OBJECTIVE: Our goal was to summarize the relationship between vein diameters, reflux characteristics, and clinical severity in consecutive patients with chronic venous insufficiency (CVI) in Northwest China. METHODS: We evaluated 531 consecutive patients with CVI (249 women) who presented to the Department of Ultrasound of Xijing Hospital from September 2017 to July 2019. Reflux times and the mean diameters of the great saphenous, the small saphenous, and the calf perforator veins based on duplex ultrasound scans obtained in the standing position were recorded. Venous-specific assessment tools-the Heaviness, Achiness, Swelling, Throbbing, Itching (HASTI) score, the Venous Clinical Severity Score (VCSS), and the Clinical, Etiological, Anatomical, Pathophysiological (CEAP) class-were analyzed. Regression analysis was used to investigate the relationship between the clinical scores, vein diameters, and reflux times. A P value of less than .05 was considered statistically significant. RESULTS: We analyzed 531 consecutive patients with 728 limbs. The mean age was 55.24 ± 11.38 years; the mean body mass index (BMI) was 24.75 ± 3.49 kg/m2. Three hundred thirty-four patients (62.9%) presented with unilateral limb findings and 197 (37.1%), with bilateral limb involvement. No significant changes were noted in age and BMI across CEAP classes (F = 2.322 and F = 3.917, respectively; P > .05 for both). Both the HASTI score (r2 = 0.8741; P < .001) and the VCSS (r2 = 0.9257; P < .001) correlated with the CEAP class. The HASTI score strongly correlated with the mean diameters of the great saphenous and small saphenous veins (r2 = 0.9252, r2 = 0.6304, respectively; P < .001 for both) similarly to VCSS (r2 = 0.9396, r2 = 0.7195, respectively; P < .001 for both). The HASTI score and VCSS correlated equally with the mean diameters of the calf perforator veins (r = 0.7773 and r = 0.7781, respectively; P < .001 for both). In those with C6, both great saphenous vein (F = 4.608; P < .001) and small saphenous vein reflux times (F = 14.97; P < .001) were significantly higher than those in C1. Both the HASTI score and VCSS strongly associated with the reflux times of the great saphenous (r2 = 0.7706 and r2 = 0.8181, respectively; P < .001 for both) and small saphenous veins (r2 = 0.6470 and r2 = 0.7865, respectively; P < .001 for both). CONCLUSIONS: This analysis is one of the few epidemiologic studies of patients with CVI in Northwest China. Age and BMI did not correlate with CEAP class. Both the HASTI score and VCSS correlated strongly with the CEAP classification; vein diameters and reflux time in both the great saphenous vein and the small saphenous vein, indicating the validity of these outcome tools to venous hemodynamics and to CVI in general.

Downregulation of RKIP promotes radioresistance of nasopharyngeal carcinoma by activating NRF2/NQO1 axis via downregulating miR-450b-5p.

Dysregulation of RKIP and NRF2 has been widely involved in the therapy resistance of multiple malignances, however, their relation and the corresponding mechanisms, especially in radiation response, have not been elucidated. In this study, we revealed that RKIP could negatively regulate the expression of NRF2 in nasopharyngeal carcinoma (NPC) cells. Depletion or ectopic expression of NRF2 countered the pro- or anti- radioresistant effects of RKIP knockdown or overexpression on NPC cells, respectively, both in vitro and in vivo. Furthermore, our results indicated that NQO1 was positively regulated by NRF2 and served as the downstream effector of RKIP/NRF2 axis in regulation of NPC radioresistance. Mechanistically, miR-450b-5p, being positively regulated by RKIP in NPC cells, could sensitize NPC cells to irradiation by directly targeting and suppressing the level of NRF2. Besides, we analyzed the level of aforementioned molecules in NPC tissues. The results indicated that RKIP was significantly downregulated, NRF2 and NQO1 were notably upregulated in NPC tissues compared with in normal nasopharyngeal mucosa (NNM) tissues. Furthermore, RKIP and miR-450b-5p were remarkably lower, yet NRF2 and NQO1 were notably higher, in radioresistant NPC tissues relative to in radiosensitive NPC tissues. Consistent with the pattern in NPC cells, the RKIP/miR-450b-5p/NRF2/NQO1 axis was significantly correlated in NPC tissues. Downregulation of RKIP and miR-450b-5p, and upregulation of NRF2 and NQO1, positively correlated to malignant pathological parameters such as primary T stage, Lymph node (N) metastasis, and TNM stage. Finally, RKIP and miR-450b-5p served as favorable prognostic indicators, and NRF2 and NQO1 acted as unfavorable prognostic biomarkers in patients with NPC. Collectively, our outcomes reveal that RKIP downregulation promotes radioresistance of NPC by downregulating miR-450b-5p and subsequently upregulating and activating NRF2 and NQO1, highlighting RKIP/miR-450b-5p/NRF2/NQO1 axis as a potential therapeutic target for improving the radiosensitivity of NPC.

Continuous growth phenomenon for direct synthesis of monodisperse water-soluble iron oxide nanoparticles with extraordinarily high relaxivity.

The direct synthesis of highly water-soluble nanoparticles has attracted intensive interest, but systematic size control has not been reported. Here, we developed a general method for synthesizing monodisperse water-soluble iron oxide nanoparticles with nanometer-scale size increments from 4 nm to 13 nm in a single reaction. Precise size control was achieved by continuous growth in an amphiphilic solvent, diethylene glycol (DEG), where the growth step was separated from the nucleation step by sequential addition of a reactant. There was only one reactant in the synthesis and no need for additional capping agents and reducing agents. This study reveals the "living growth" character of iron oxide nanoparticles synthesised in an amphiphilic solvent. The synthetic method shows high reproducibility. The as-prepared iron oxide nanoparticles are extremely water soluble without any surface modification. Surprisingly, the synthesized 9 nm iron oxide nanoparticles exhibit extremely high transversal and longitudinal relaxivities of 425 mM-1 s-1 and 32 mM-1 s-1 respectively, which is among the highest transversal relaxivity in the literature for sub-10 nm spherical nanoparticles. This study will not only shed light on the continuous growth phenomenon of iron oxide nanoparticles in an amphiphilic solvent, but could also stimulate the synthesis and application of iron oxide nanoparticles. The continuous growth method could be further extended to other materials for the controlled synthesis of water-soluble nanoparticles.

[Application of superb microvascular imaging and contrast enhanced ultrasound in the differential diagnosis of thyroid nodules].

OBJECTIVE: To compare the clinical value of superb microvascular imaging (SMI) and contrast enhanced ultrasound (CEUS) in the differential diagnosis of thyroid nodules, and to further study whether the combination of SMI and/or CEUS with thyroid imaging reporting and data system (TI-RADS) can improve the diagnostic value of TI-RADS.
 Methods: SMI, CEUS, TI-RADS, TI-RADS combined with CEUS, TI-RADS combined with SMI, TI-RADS combined with SMI and CEUS were used to differentiate thyroid nodules. TI-RADS 4b, 5 categories in two-dimensional ultrasound, hypoenhancement in CEUS, and peripheral blood flow with penetrating vessels in SMI were considered malignant signs. The diagnostic efficacy of these methods was compared according to post-operative pathology or fine needle aspiration cytology.
 Results: A total of 237 patients with 296 thyroid nodules were included in the study. The sensitivity, specificity and accuracy for TI-RADS were 78.4%, 77.7% and 78.0% respectively, 75.5%, 86.6% and 81.4% in SMI, 82.0%, 88.5% and 85.5% in CEUS, 92.1%, 90.4% and 91.2% in TI-RADS combined with SMI and CEUS. Area under the receiver operating characteristic curve (AUROC) showed no significant difference between SMI, CEUS, TI-RADS (χ2=4.29, P=0.117). AUROC of both TI-RADS combined with CEUS (χ2=39.62, P<0.001), TI-RADS combined with SMI (χ2=36.61, P<0.001) were higher than TI-RADS. AUROC of TI-RADS combined with SMI and CEUS was higher than TI-RADS combined with SMI (χ2=4.50, P=0.033) or TI-RADS combined with CEUS (χ2=4.24, P=0.039).
 Conclusion: Peripheral blood flow with penetrating vessels in SMI possesses high diagnostic value in differentiating thyroid cancer. The combination of SMI and/or CEUS with TI-RADS can be used to improve the diagnostic efficacy of TI-RADS in differential diagnosis of thyroid nodules.

Assessment of Copper Nanoclusters for Accurate in Vivo Tumor Imaging and Potential for Translation.

Nanoparticles have been widely used for preclinical cancer imaging. However, their successful clinical translation is largely hampered by potential toxicity, unsatisfactory detection of malignancy at early stages, inaccurate diagnosis of tumor biomarkers, and histology for imaging-guided treatment. Herein, a targeted copper nanocluster (CuNC) is reported with high potential to address these challenges for future translation. Its ultrasmall structure enables efficient renal/bowel clearance, minimized off-target effects in nontargeted organs, and low nonspecific tumor retention. The pH-dependent in vivo dissolution of CuNCs affords minimal toxicity and potentially selective drug delivery to tumors. The intrinsic radiolabeling through the direct addition of 64Cu to CuNC (64Cu-CuNCs-FC131) synthesis offers high specific activity for sensitive and accurate detection of CXCR4 via FC131-directed targeting in novel triple negative breast cancer (TNBC) patient-derived xenograft mouse models and human TNBC tissues. In summary, this study not only reveals the potential of CXCR4-targeted 64Cu-CuNCs for TNBC imaging in clinical settings, but also provides a useful strategy to design and assess the translational potential of nanoparticles for cancer theranostics.