Active Anchoring Stimuli-Responsive Nano-Craft to Relieve Pulmonary Vasoconstriction by Targeting Smooth Muscle Cell for Hypoxic Pulmonary Hypertension Treatment.

Recently, nanotechnology-based drug delivery platforms in treating pulmonary arterial hypertension (PAH) have gradually emerged. However, large mechanical stress and shear stress in blood vessels greatly affect the retention of nanopreparative materials at lesion sites, severely limiting nanotechnology-based drug delivery. Herein, a stimuli-responsive nanocraft is rationally designed by actively anchoring E-selectin overexpressed on pulmonary arterial endothelial cells (PAECs), under hypoxic conditions, allowing effective accumulation and retention of the drug at the lesion site. Briefly, a nitrobenzene group is incorporated into the framework of a nanocarrier, and then it is simultaneously linked with chitosan. Additionally, the surface of the nanocarrier with sialic acid (SA) and encapsulated the clinically used drug ambrisentan (Am), which enables the anchoring of E-selectin and subsequent drug delivery is modifed. This system facilitates intercellular transport to pulmonary artery smooth muscle cells (PASMCs) when targeting PAECs and specifically responds to a reductive hypoxic microenvironment with elevated nitroreductase in PASMCs. Moreover, compared with free Am, nanoencapsulation and SA-PEG2000-NH2 prolong the blood circulation time, achieving better therapeutic outcomes in preventing vascular remodeling and reversing systolic dysfunction. The originality and contribution of this work reveal the promising value of this pulmonary arterial anchoring stimuli-responsive nanocraft as a novel therapeutic strategy for satisfactory PAH treatment.

Low-Intensity Focused Ultrasound Targeted Microbubble Destruction Enhanced Paclitaxel Sensitivity by Decreasing Autophagy in Paclitaxel-Resistant Ovarian Cancer.

Ultrasound targeted microbubble destruction (UTMD) was introduced as a promising method to improve anti-tumor therapeutic efficacy, while minimizing side effects to healthy tissues. Nevertheless, the acoustical phenomenon behind the UTMD as well as the exact mechanisms of autophagy action involved in the increased anti-cancer response are still not fully understood. Therefore, we examined the drug resistance-reversing effects of low-intensity focused ultrasound with microbubble (LIFU+MB) in paclitaxel (PTX)-resistant ovarian cancer cells. Cell viability was evaluated using CCK8 (Cell Counting Kit-8), apoptosis was detected by flow cytometry, quantitative real-time PCR and Western blot were used to detect the expressions of mRNA and protein, and autophagy was observed by transmission electron microscopy (TEM). We revealed that the level of autophagy was increased (p < 0.05) in PTX-resistant ovarian cancer cells. Treatment of LIFU+MB combined with PTX can notably inhibit proliferation as well as increase apoptosis (p < 0.01) in drug-resistant cells. We proposed that LIFU+MB might affect the sensitivity of ovarian cancer cells to PTX by modulating autophagy. To verify the hypothesis, we analyzed the autophagy level of drug-resistant cells after the treatment of LIFU+MB and found that autophagy was significantly inhibited. Altogether, our findings demonstrated that LIFU+MB could reverse PTX resistance in ovarian cancer via inhibiting autophagy, which provides a novel strategy to improve chemosensitivity in ovarian cancer.

Self-preparation system using glucose oxidase-inspired nitroreductase amplification for cascade-responsive drug release and multidrug resistance reversion.

Early antitumor therapy is an important determinant of survival in patients with cancer. Utilization of specific pathological states, such as hypoxia, greatly promotes the development of intelligent drug delivery systems (DDSs) for targeted antitumor therapy. However, a slight decrease in oxygen levels in early-stage tumors is not sufficient to trigger hypoxia-responsive drug release. Nitroreductase (NTR) is overexpressed in bioreductive hypoxic cancers, and its expression level has been verified to be directly related to hypoxic status. Herein, using glucose oxidase (GOx) as an O2-consuming agent to exacerbate hypoxia, a cascade strategy of GOx-induced overexpression of NTR and amplified NTR-catalyzed release was proposed for early antitumor therapy. Briefly, NTR-sensitive p-nitrobenzyl chloroformate (PNZ-Cl) was adopted to conjugate with the polysaccharide chitosan (CS) and self-assemble into CS-PNZ-Cl micelles. These polymer micelles possess the dual abilities to specifically immobilize GOx and load mitoxantrone (MIT) to form the NTR-responsive nanocascade reactor GOx/MIT@CS-PNZ-Cl. First, as a "key", tumor hypoxia triggers the initial release of GOx, which serves as the O2-consuming agent when catalyzing its reaction with glucose, which is accompanied by H2O2 production. Depleted oxygen levels facilitate the expression of NTR, which in turn amplifies the capacity of the nanocascade reactor to decompose into secondary micelles for enhanced intratumoral permeation. GOx-inspired NTR amplification further elicits MIT release, realizing a synergistic "domino effect" cascade. In addition, upregulated H2O2 has been shown to effectively reverse GSH-mediated MIT resistance, reaching the superior tumor inhibition rate of 93.08%. This GOx-based NTR-responsive nanocascade reactor provides amplification of the bioreductive hypoxic tumor microenvironment for early antitumor therapy.

Quercetin Covalently Linked Lipid Nanoparticles: Multifaceted Killing Effect on Tumor Cells.

The encapsulation of hydrophobic drugs is a problem that many researchers are working on. The goal of this study is to achieve the delivery of hydrophobic drugs by means of prodrugs and nanoformulations for a stronger tumor cell-killing effect and explore related killing mechanisms. Lipophilic quercetin (Qu) was covalently linked to glyceryl caprylate-caprate (Gcc) via disulfide bonds-containing 3,3'-dithiodipropionic acid (DTPA) to synthesize novel lipid Qu-SS-Gcc. Qu-SS-Gcc lipid nanoparticles (Qu-SS-Gcc LNPs) were fabricated using the solvent diffusion technique. The intracellular release of Qu by cleavage of nanocarriers was determined by liquid chromatography and compared with the uptake of free Qu. Detection methods, such as fluorescent quantitation, flow cytometry, and western blot were applied to explore the action mechanism induced by Qu. It was revealed that Qu-SS-Gcc LNPs could be cleaved by the high concentrations of reduction molecules in MCF-7/ADR (human multidrug-resistant breast cancer) cells, followed by the release of Qu. The intracellular Qu content produced by dissociation of Qu-SS-Gcc LNPs was higher than that produced by internalization of free Qu. The resulting release of Qu exerted superior cell-killing effects on MCF-7/ADR cells, such as P-gp inhibition by binding to P-gp binding sites, blocking the cell cycle in the G2 phase, and causing cell apoptosis and autophagy. Moreover, it was revealed autophagy triggered by a low concentration of Qu-SS-Gcc LNPs was beneficial to cell survival, while at a higher concentration, it acted as a cell killer. Qu-SS-Gcc LNPs can realize massive accumulation of Qu in tumor cells and exert a multifaceted killing effect on tumor cells, which is a reference for the delivery of hydrophobic drugs.

Calcium Supplement by Tetracycline guided amorphous Calcium Carbonate potentiates Osteoblast promotion for Synergetic Osteoporosis Therapy.

Background: The calcium supplement is a clinically approved approach for osteoporosis therapy but usually requires a large dosage without targetability and with poor outcome. This modality is not fully explored in current osteoporosis therapy due to the lack of proper calcium supplement carrier. Methods: In this study, we constructed a tetracycline (Tc) modified and simvastatin (Sim) loaded phospholipid-amorphous calcium carbonate (ACC) hybrid nanoparticle (Tc/ACC/Sim). Results: The resulted Tc/ACC/Sim was able to enhance its accumulation at the osteoporosis site. Most importantly, the combination of calcium supplement and Sim offered synergetic osteoblast promotion therapy of osteoporosis with advanced performance than non-targeted system or mono therapy. Conclusion: This platform provides an alternative approach to stimulate bone formation by synergetic promotion of osteoblast differentiation using calcium supplement and Sim.

Chitosan-modified lipid nanodrug delivery system for the targeted and responsive treatment of ulcerative colitis.

Targeted and sensitive drug release at the colitis site is critical for the effective therapy of ulcerative colitis and reduction of side effects from the drug. Herein, we used 3,3'-dithiodipropionic acid (DTPA) to covalently link quercetin (Qu) and glyceryl caprylate-caprate (Gcc) via ester bonds to prepare Qu-SS-Gcc lipid nanoparticles (Qu-SS-Gcc LNPs). Dexamethasone (Dex) was used as a model drug, and chitosan (CSO) was modified on the surface of Qu-SS-Gcc LNPs to obtain CSO-modified Dex-loaded Qu-SS-Gcc LNPs (CSO/Dex/LNPs). The encapsulation efficiency and drug loading of CSO/Dex/LNPs were 93.1 % and 8.1 %, respectively. The in vitro release results showed that CSO/Dex/LNPs had esterase-responsive characteristics and could release the drug rapidly in esterase-containing artificial intestinal fluid. A human colorectal adenocarcinoma cell (Caco-2) monolayer was used as the intestinal cell barrier model. Transmembrane resistance measurements and permeation experiments showed that CSO/Dex/LNPs had a protective effect on the lipopolysaccharide (LPS)-stimulated Caco-2 cell monolayer and increased the expression of E-cadherin in LPS-stimulated Caco-2 cells. Moreover, CSO/Dex/LNPs could significantly reduce the expression of the inflammatory factors TNF-α, IL-6 and NO in LPS-stimulated RAW 264.7 cells. The ulcerative colitis mouse model was constructed by using C57BL/6 mice. The in vivo distribution results showed that CSO/Dex/LNPs had colon-targeting effects and strong retention ability in the colons of mice with colitis. The results also showed that CSO/Dex/LNPs had better anti-inflammatory effects than free Dex, which could reduce colonic atrophy, reduce histomorphological changes and increase the expression of E-cadherin in the colon. Furthermore, the expression levels of TNF-α, IL-6 and NO in the CSO/Dex/LNP-treated group were 37.4 %, 35.5 % and 33.2 % of those in mice with colitis, respectively.

[Sonographic features and diagnostic analysis of breast granulomatous diseases: a report of 32 cases].

OBJECTIVE: To evaluate the sonographic features of different pathological types of breast granulomatous diseases and analyze the feasibility of ultrasonic diagnosis. METHODS: A total of 32 patients with different pathological types of breast granulomatous diseases were recruited. Their clinical and sonographic findings were retrospectively reviewed. There were granulomatous mastitis (n = 12), breast xanthogranuloma (n = 5), lipogranuloma (n = 2), foreign body granuloma (n = 1) and nonspecific granulation hyperplasia (n = 12). RESULTS: Based on major sonographic appearances, they were divided into 4 patterns of tubular, mass, diffuse and cystic mass. In 12 patients with granulomatous mastitis and 12 patients with nonspecific granulation hyperplasia, the major sonographic appearance was of tubular pattern (n = 6, 5), followed by mass pattern (n = 4, 5) and diffuse pattern (n = 2, 2). Five patients with breast xanthogranuloma and 1 patient with foreign body granuloma all showed mass pattern. In 2 patients with lipogranuloma, one was of mass pattern and another cystic pattern. In patients with granulomatous mastitis and patients with nonspecific granulation hyperplasia, it showed a high diagnostic reliability of ultrasound. The ratio of inflammatory lesion as the first sonographic diagnosis was 10/12 and 8/12 respectively and ultrasonic BI-RADS 4b or above both only 1/12. However, the ratio of sonographic imaging in patients with xanthogranuloma and Lipogranuloma mimic breast cancer, in which ultrasonic score as breast imaging-reporting and data system (BI-RADS) 4b or above was 4/5 and 1/1 respectively. CONCLUSIONS: Ultrasound is valuable in evaluating the lesions in patients with granulomatous mastitis and nonspecific granulation hyperplasia. However a definite diagnosis is still dependent on histopathology.

[Sonographic and computed tomography features of hepatic angiomyolipoma].

OBJECTIVE: To investigate the sonographic and computed tomography (CT) features of hepatic angiomyolipoma (HAML). METHODS: Sonographic and CT findings were analyzed in 12 patients (9 females and 3 males) with pathologically proved HAML. The size, margin, location, gray scale, and color Doppler flow imaging characteristics were observed. RESULTS: HAML was located correctly with ultrasound in all patients. The sonographic features of 12 HAML included regular shape, clear margin, and three type of echoes including homogeneous hyperechoes (n=5), heterogeneous internal echoes (n=5), or homogeneous hypoechoes (n=2). The arterial flow signal was detected in two HAML. The CT findings included adipose density (n=3), soft tissue density (n=3), and mixed density (n=6). The sonographic and CT findings were correlated with the composition and distribution of fat, vessels, and smooth muscle tissue. CONCLUSIONS: Fatty tissues within HAML shows typical imaging findings. The ultrasonographic and CT have their own advantages in detecting the fatty tissue inside HAML, and therefore a combination of these two techniques may increase the diagnostic accuracy of HAML.

[Application values of four risk of malignancy indices in the preoperative evaluation of patients with adnexal masses].

OBJECTIVE: To evaluate the diagnostic values of four risk of malignancy indices (RMI) for malignant adnexal masses. METHODS: The data of 223 women with adnexal masses admitted to the Department of Obstetrics and Gynecology of Peking Union Medical College Hospital for surgical exploration between June 2008 and December 2008 were retrospectively analyzed. The sensitivity, specificity, positive predictive value and negative predictive value of RMI1, RMI2, RMI3, and RMI4 in the diagnosis of malignant adnexal masses were calculated. RESULTS: When the cutoff levels of RMI1, RMI2, RMI3 were set at 200 and RMI4 at 450, the sensitivities for diagnosing malignant adnexal masses ranged 59.0%-67.2%, the specificities ranged 94.4%-96.9 %, the positive predictive values ranged 82.0%-87.8%, and the negative predictive values ranged 90.9%-92.6%. The Youdens indexes (YI) of RMI1, RMI2, RMI3, and RMI4 were 0.559,0.606,0.576, and 0.559, respectively. RMI2 was significantly different from RMI1 (P=0.000), RMI3 (P=0.008), and RMI4 (P=0.000) in terms of diagnostic efficiency. RMI1, RMI2, RMI3, and RMI4 at a cutoff level of 75.688.679.1, 177.2 respectively, according to ROC curves, yielded sensitivities of 77.8%-82.5%, specificities of 84.6%-90.1%, positive predictive values of 69.0%-75.4%, and negative predictive values of 90.9%-92.6%; the relevant YI of RMI1, RMI2, RMI3, and RMI4 were 0.635, 0.665, 0.651 and 0.705, respectively. Under this cutoff level, the difference between RMI1, RMI2, RMI3, and RMI4 in diagnosing malignancy had no statistic significant. The primary histological types arising false negative were early stage epithelial ovarian cancer and non-epithelial ovarian cancer. The primary histological types arising false positive were endometriosis masses and degenerative sex cord-stromal tumor. CONCLUSIONS: RMIs are useful indices for the differentiation between benign and malignant pelvic diseases. Meanwhile, their cutoff levels for Chinese populations need further study.

[Possibility of PTEN expression on predicting pathologic risk factors of endometrioid adenocarcinoma before operation].

BACKGROUND & OBJECTIVE: Fully estimating pathologic risk factors is important for selecting operation and predicting prognosis for endometrioid adenocarcinoma. Phosphatase and tension homology deleted on chromosome ten (PTEN), taken as the housekeeping gene of endometrium, has the highest mutation rate in endometrioid adenocarcinoma. This study was to investigate the effect of PTEN on predicting pathologic risk factors of endometrioid adenocarcinoma before operation. METHODS: Clinicopathologic data of 107 patients with endometrioid adenocarcinoma were retrospectively analyzed. The expression of PTEN was detected by SP immunohistochemistry. Correlations of PTEN to high risk factors, such as differentiation, myometrium invasion, and lymphatic metastasis, were analyzed. RESULTS: Deletion rate of PTEN was 56.1% in the 107 endometrioid adenocarcinoma patients. PTEN expression had no correlations to histological differentiation (P=0.695), myometrium invasion (P=0.921), lymphatic metastasis (P=0.682), surgical stage (P=0.750), estrogen receptor (P=0.281), and progestin receptor (P=0.260). CONCLUSION: Detection of PTEN can't predict the high risk factors of endometrioid adenocarcinoma before operation.