Effect of food on the pharmacokinetics and safety profiles of a new PARP inhibitor fuzuloparib capsules in healthy volunteers.

PURPOSE: This study assessed effect of food on pharmacokinetics (PK) and safety of fuzuloparib capsules. METHODS: A randomized, open-label, two-cycle, two-sequence, crossover clinical trial was conducted. 20 subjects were randomly assigned to 2 groups at a 1:1 ratio. The first group subjects were orally administered 150 mg fuzuloparib capsules under fasting condition in first dosing cycle. The same dose of fuzuloparib capsules were taken under postprandial state after a 7-day washout period. The second group was reversed. 3 ml whole blood was collected at each blood collection point until 72 h post dose. PK parameters were calculated. Furthermore, safety assessment was performed. RESULTS: The time to maximum concentration (Tmax) was prolonged to 3 h and maximum concentration (Cmax) decreased by 18.6% on high-fat diets. 90% confidence intervals (CIs) of geometric mean ratios (GMRs) for Cmax, area under the concentration-time curve from time zero to time t (AUC0-t), and area under the concentration-time curve extrapolated to infinity (AUC0-∞) after high-fat meal were 71.6-92.6%, 81.7-102.7% and 81.6-102.5%, respectively. All treatment-emergent adverse events (TEAEs) were grade 1; No serious adverse events (SAEs), serious unexpected suspected adverse reaction (SUSAR) or deaths were reported. CONCLUSION: Food decreased the absorption rate and slowed time to peak exposure of fuzuloparib capsules, without impact on absorption extent. Dosing with food was found to be safe for fuzuloparib capsules in this study. CLINICAL TRIAL REGISTRATION: This study was registered with chinadrugtrials.org.cn (identifier: CTR20221498).

mRNA-responsive two-in-one nanodrug for enhanced anti-tumor chemo-gene therapy.

The combination of chemotherapy and gene therapy holds great promise for the treatment and eradication of tumors. However, due to significant differences in physicochemical properties between chemotherapeutic agents and functional nucleic acid drugs, direct integration into a single nano-agent is hindered, impeding the design and construction of an effective co-delivery nano-platform for synergistic anti-tumor treatments. In this study, we have developed an mRNA-responsive two-in-one nano-drug for effective anti-tumor therapy by the direct self-assembly of 2'-fluoro-substituted antisense DNA against P-glycoprotein (2'F-DNA) and chemo drug paclitaxel (PTX). The 2'-fluoro modification of DNA could significantly increase the interaction between the therapeutic nucleic acid and the chemotherapeutic drug, promoting the successful formation of 2'F-DNA/PTX nanospheres (2'F-DNA/PTX NSs). Due to the one-step self-assembly process without additional carrier materials, the prepared 2'F-DNA/PTX NSs exhibited considerable loading efficiency and bioavailability of PTX. In the presence of endogenous P-glycoprotein mRNA, the 2'F-DNA/PTX NSs were disassembled. The released 2'F-DNA could down-regulate the expression of P-glycoprotein, which decreased the multidrug resistance of tumor cells and enhanced the chemotherapy effect caused by PTX. In this way, the 2'F-DNA/PTX NSs could synergistically induce the apoptosis of tumor cells and realize the combined anti-tumor therapy. This strategy might provide a new tool to explore functional intracellular co-delivery nano-systems with high bioavailability and exhibit potential promising in the applications of accurate diagnosis and treatment of tumors.

Benign thyroid nodules classified as ACR TI-RADS 4 or 5: Imaging and histological features.

BACKGROUND: American College of Radiology Thyroid Imaging Reporting and Data System (ACR TI-RADS) being most widely applied in clinical practice, there is an overlap in US imaging manifestations between benign and malignant thyroid nodules. OBJECTIVES: To analyze the imaging and histological characteristics of pathological benign thyroid nodules categorized as American College of Radiology Thyroid Imaging Reporting and Data System (ACR TI-RADS) 4 or 5, and to explore the correlation between the suspicious sonographic signs resulting in the misdiagnoses and the histopathological features. MATERIALS AND METHODS: Overall, 227 benign thyroid nodules (215 patients) in ACR TI-RADS 4 or 5 sampled through surgical excision were analyzed between December 2020 and August 2022. We retrospectively reread the ultrasound (US) images of the pathological discordant cases, after which we performed a systematic analysis focusing on the histopathological characteristics of thyroid lesions and recorded the findings. Qualitative US features and pathological significance of the thyroid nodules were analyzed using the chi-square and Fisher's exact tests. RESULTS: The pathological type of 227 thyroid nodules (n = 103 in ACR TI-RADS 4 and n = 124 in ACR TI-RADS 5) was nodular goiter together with other histopathological features, namely, fibrosis (n = 103, 45.4 %), calcification (n = 70, 30.8 %), adenomatous hyperplasia (n = 31, 13.7 %), follicular epithelial hyperplasia (n = 23, 10.1 %), Hashimoto's thyroiditis (n = 18, 7.9 %), and cystic degeneration (n = 16, 7.1 %). Fibrosis was the most common histopathological feature in both ACR TI-RADS 4 (n = 42, 40.8 %) and 5 (n = 61, 49.2 %) categories of benign thyroid nodules. Thyroid nodules with fibrosis demonstrated sonographic features of "taller than wide" (p < 0.05), while lesions with follicular epithelial hyperplasia were likely to be detected with irregular and/or lobulated margins and very hypoechoic on US (p < 0.05 for both). CONCLUSION: Benign thyroid nodules with histopathological findings such as fibrosis are associated with suspicious US features, which may give inappropriately higher TIRADS stratification.

Diagnostic performance of ACR-TIRADS combined with superb microvascular imaging for differential diagnosis of mummified thyroid nodules and papillary thyroid carcinomas.

Objective: The aim was to investigate the ability of superb microvascular imaging (SMI) to improve the differential diagnosis of mummified thyroid nodules (MTNs) and papillary thyroid carcinomas (PTCs) using the 2017 American College of Radiology Thyroid Imaging Reporting and Data System (ACR-TIRADS). Materials and methods: We enrolled 110 cases of MTNs and 110 cases of PTCs confirmed by fine needle aspiration (FNA) or surgery. Conventional ultrasound (US) and the quantity of microvessels detected by SMI were analyzed for all nodules. Thyroid nodules were initially categorized by ACR-TIRADS based on US imaging features and then reclassified based on ACR-TIRADS combined with SMI blood-flow grade (SMI-TIRADS). We compared the diagnostic performances of ACR-TIRADS and SMI-TIRADS by receiver operating characteristic curve, sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV). Results: US-detected margin, shape, and echogenic foci differed between MTNs and PTCs (P < 0.05). The SMI blood-flow grade was significantly greater in PTCs compared with MTNs (Χ2 = 158.78, P < 0.05). There was no significant difference in ACR-TIRADS indicators between MTNs and PTCs (Χ2 = 1.585, P = 0.453); however, reclassification by SMI-TIRADS showed significant differences between the groups (Χ2 = 129.521, P < 0.001). The area under the curve was significantly lower for ACR-TIRADS compared with SMI-TIRADS (0.517 vs 0.887, P < 0.05). SMI-TIRADS had significantly higher diagnostic value for distinguishing MTNs and PTCs than ACR-TIRADS (sensitivity: 91.82% vs 74.55%, P < 0.05; specificity: 84.55% vs 21.82%, P < 0.05; accuracy: 88.18% vs 48.18%, P < 0.05; PPV: 85.59% vs 48.81%, P < 0.05; and NPV: 91.18% vs 46.15%, P < 0.05). Conclusion: The detection of microvascular flow and large vessels in thyroid nodules by SMI resulted in high diagnostic specificity and sensitivity. ACR-TIRADS combined with SMI could effectively distinguish between MTNs and PTCs, to avoid unnecessary FNA or surgical excision.

Postoperative quality of life in patients with ankylosing spondylitis and thoracolumbar kyphosis: risk factors and personalized sagittal reconstruction strategy.

OBJECTIVE: The aim of this study was to investigate the factors affecting postoperative quality of life in patients with ankylosing spondylitis (AS) and thoracolumbar kyphosis (TLK), and establish a personalized sagittal reconstruction strategy. METHODS: Patients with AS and TLK who underwent pedicle subtraction osteotomy (PSO) from February 2009 to May 2019 were retrospectively included. Quality of life and spinal sagittal radiographic parameters were collected before surgery and at the last follow-up. Patients were divided into two groups based on the attainment of minimal clinically important difference (MCID) on the Bath Ankylosing Spondylitis Functional Index and Oswestry Disability Index. Comparisons of radiographic parameters and clinical outcomes were conducted between and within groups. Regression analysis was used to identify the risk factors within the missing MCID cohort. Sagittal reconstruction equations were established using the pelvic incidence (PI) and thoracic inlet angle (TIA) in the reached MCID cohort. RESULTS: The study comprised 82 participants. Significant improvements were observed in most radiographic parameters and all quality-of-life indicators during the final follow-up compared with the preoperative measures (p < 0.05). Factors including cervical lordosis (CL) ≥ 18° (OR 9.75, 95% CI 2.26-58.01, p = 0.005), chin-brow vertical angle (CBVA) ≥ 25° (OR 14.7, 95% CI 3.29-91.21, p = 0.001), and pelvic tilt (PT) ≥ 33° (OR 21.77, 95% CI 5.92-103.44, p < 0.001) independently correlated with a failure to attain MCID (p < 0.05). Sagittal realignment targets were constructed as follows: sacral slope (SS) = 0.84 PI - 17.4° (R2 = 0.81, p < 0.001), thoracic kyphosis (TK) = 0.51 PI + 10.8° (R2 = 0.46, p = 0.002), neck tilt (NT) = 0.52 TIA - 5.8° (R2 = 0.49, p < 0.001), and T1 slope (T1S) = 0.48 TIA + 5.8° (R2 = 0.45, p = 0.002). CONCLUSIONS: PSO proved efficacious in treating AS complicated by TLK, yielding favorable outcomes. CBVA ≥ 25°, CL ≥ 18°, and PT ≥ 33° were the primary factors affecting postoperative quality of life in patients with AS. The personalized sagittal reconstruction strategy in this study focused on the subjective sensations and daily needs of patients with AS, which were delineated by the equations SS = 0.84 PI - 17.4°, TK = 0.51 PI + 10.8°, NT = 0.52 TIA - 5.8°, and T1S = 0.48 TIA + 5.8°.

Ultrasound-Guided Percutaneous Transhepatic Cholangioscopic Lithotripsy for the Treatment of Common Bile Duct Stones and Analysis of Risk Factors for Recurrence.

BACKGROUND: As a minimally invasive treatment for common bile duct (CBD) stones, ultrasound-guided percutaneous transhepatic cholangioscopic lithotripsy (PTCSL) is gaining attention and recognition from the medical community. METHODS: A retrospective analysis was conducted on patients with CBD stones treated in our hospital from January 2016 to April 2022. Patients were divided into three groups: 77 treated with PTCSL, 93 with endoscopic retrograde cholangiopancreatography (ERCP), and 103 with laparoscopic common bile duct exploration (LCBDE). Their clinical data, perioperative indicators, and complications were analyzed comparatively. Then, risk factors for the post-PTCSL recurrence of CBD stones were analyzed by logistic regressions. Finally, the receiver operating characteristic curve was drawn. RESULTS: All perioperative indicators of the PTCSL group were better than the LCBDE group (P < 0.001). The incidences of cholangitis, hemobilia, and incisional infection after surgery were lower in the PTCSL group than in the LCBDE group (P < 0.05). Pancreatitis, reflux esophagitis, and papillary stenosis occurred less frequently in the PTCSL group than in the ERCP group (P < 0.05). Logistic regression analysis indicated that gallstones and family history were independent risk factors. The AUC for recurrent CBD stones predicted by multi-indicators was 0.895 (95% CI 0.792-0.999, P < 0.001) with a sensitivity of 96.7% and specificity of 68.8%. CONCLUSIONS: Ultrasound-guided PTCSL is a safe and effective treatment for CBD stones. Patients recovered quickly with fewer postoperative complications. It can be a first-line treatment for CBD stones. Gallstones and family history are independent risk factors for recurrent CBD stones, which provide a reference for clinicians in identifying the high-risk population needing close follow-up.

Autophagy-amplifying nanoparticles evoke immunogenic cell death combined with anti-PD-1/PD-L1 for residual tumors immunotherapy after RFA.

Incomplete radiofrequency ablation (IRFA) triggers mild protective autophagy in residual tumor cells and results in an immunosuppressive microenvironment. This accelerates the recurrence of residual tumors and causes resistance to anti-PD-1/PDL1 therapy, which bringing a great clinical challenge in residual tumors immunotherapy. Mild autophagy activation can promote cancer cell survival while further amplification of autophagy contributes to immunogenic cell death (ICD). To this regard, we constructed active targeting zeolitic imidazolate framework-8 (ZIF-8) nanoparticles (NPs) loaded with STF62247 or both STF62247 and BMS202, namely STF62247@ZIF-8/PEG-FA (SZP) or STF62247-BMS202@ZIF-8/PEG-FA (SBZP) NPs. We found that SZP NPs inhibited proliferation and stimulated apoptosis of residual tumor cells exposed to sublethal heat stress in an autophagy-dependent manner. Further results discovered that SZP NPs could amplify autophagy in residual tumor cells and evoke their ICD, which dramatically boosted the maturation of dendritic cells (DCs). Through vaccination experiments, we found for the first time that vaccination with heat + SZP treatment could efficiently suppress the growth of new tumors and establish long-term immunological memory. Furthermore, SBZP NPs could remarkably promote the ICD of residual tumor cells, obviously activate the anti-tumor immune microenvironment, and significantly inhibit the growth of residual tumors. Thus, amplified autophagy coupled with anti-PD-1/PDL1 therapy is potentially a novel strategy for treating residual tumors after IRFA.

Precise localization of microvascular invasion in hepatocellular carcinoma based on three-dimensional histology-MR image fusion: an ex vivo experimental study.

Background: Microvascular invasion (MVI) is an independent risk factor for postoperative recurrence of hepatocellular carcinoma (HCC). However, MVI cannot be detected by conventional imaging. To localize MVI precisely on magnetic resonance (MR) images, we evaluated the feasibility and accuracy of 3-dimensional (3D) histology-MR image fusion of the liver. Methods: Animal models of VX2 liver tumors were established in 10 New Zealand white rabbits under ultrasonographic guidance. The whole liver lobe containing the VX2 tumor was extracted and divided into 4 specimens, for a total of 40 specimens. MR images were obtained with a T2-weighted sequence for each specimen, and then histological images were obtained by intermittent, serial pathological sections. 3D histology-MR image fusion was performed via landmark registration in 3D Slicer software. We calculated the success rate and registration errors of image fusion, and then we located the MVI on MR images. Regarding influencing factors, we evaluated the uniformity of tissue thickness after sampling and the uniformity of tissue shrinkage after dehydration. Results: The VX2 liver tumor model was successfully established in the 10 rabbits. The incidence of MVI was 80% (8/10). 3D histology-MR image fusion was successfully performed in the 39 specimens, and the success rate was 97.5% (39/40). The average registration error was 0.44±0.15 mm. MVI was detected in 20 of the 39 successfully registered specimens, resulting in a total of 166 MVI lesions. The specific location of all MVI lesions was accurately identified on MR images using 3D histology-MR image fusion. All MVI lesions showed as slightly hyperintense on the high-resolution MR T2-weighted images. The results of the influencing factor assessment showed that the tissue thickness was uniform after sampling (P=0.38), but the rates of the tissue shrinkage was inconsistent after dehydration (P<0.001). Conclusions: 3D histology-MR image fusion of the isolated liver tumor model is feasible and accurate and allows for the successful identification of the specific location of MVI on MR images.

Endogenous dual miRNA-triggered dynamic assembly of DNA nanostructures for in-situ dual siRNA delivery.

A theranostic strategy of multiple microRNA (miRNA)-triggered in-situ delivery of small interfering RNA (siRNA) can effectively improve the precise therapy of cancer cells. Benefiting from the advantages of programmability, specific molecular recognition, easy functionalization and marked biocompatibility of DNA nanostructures, we designed a three-dimensional (3D) DNA nano-therapeutic platform for dual miRNA-triggered in-situ delivery of siRNA. The 3D DNA nanostructure (TY1Y2) was constructed based on the self-assembly of a DNA tetrahedra scaffold, two sets of Y-shaped DNA (Y1 and Y2), and EpCAM-aptamer which functionalized as the ligand molecule for the recognition of specific cancer cells. After being specifically internalized into the targeted cancer cells, TY1Y2 was triggered by two endogenous miRNAs (miR-21 and miR-122), resulting in the generation of strong fluorescence resonance energy transfer fluorescent signal for dual miRNAs imaging. Meanwhile, the therapeutic siRNAs (siSurvivin and siBcl2) could also be in-situ generated and released from TY1Y2 through the strand-displacement reactions for the synergistic gene therapy of cancer cells. This 3D DNA nanostructure integrated the specific imaging of endogenous biomarkers and the in-situ delivery of therapeutic genes into the multifunctional nanoplatform, revealing the promising applications for the diagnosis and treatment of cancer. Electronic Supplementary Material: Supplementary material is available in the online version of this article at 10.1007/s40843-022-2420-y.

Umbilical cord mesenchymal stem cells promote the repair of trochlear groove reconstruction in dogs.

Trochlear groove reconstruction (TGR) is a common treatment for patellar luxation (PL) in dogs. Nevertheless, the prognosis of TGR is poor due to the cartilage damage and secondary inflammation. To study the repair effect of canine umbilical cord mesenchymal stem cells (UC-MSCs) after TGR, 10 experimental dogs were given TGR surgery and then randomized into two groups: Treatment group (1 ml suspension allogeneic UC-MSCs (106 cells/kg) was injected into the cavum articulare on days 0, 7, and 14 after TGR); and the Model group (injected with 1 ml of physiological saline as negative control). The therapeutic effect of UC-MSCs was studied by blood routine examination, inflammatory factor index detection, double-blind knee score, histopathology, and computed tomography (CT) scans. The results showed that the total number of white blood cells and neutrophils in the model group were significantly higher than those in the treatment group on both 7 days and 21 days, postoperatively (P < 0.05); there were no significant changes in the levels of IL-6, MMP-13, and TGF-ß1 between the model group and the treatment group throughout the days of testing. The double-blind knee scores of the treatment group were significantly lower than the model group on 1st, 4th, and 5th days postoperatively (P < 0.05). The treatment group showed low-pain sensation, stable gait, and fast recovery of muscle strength in the knee score, and the wound healing of the treatment group returned to normal on the 5th day after surgery; CT scans and gross observation showed that the cartilage growth in the treatment group was faster than that in the model group. Histological observation of cases showed that fibro chondrocytes were predominantly found in the treatment group, and the distribution of chondrocytes was uneven, while the model group showed a large number of fibrous tissue hyperplasia, fissures, and unequal matrix staining. Intra-articular injection of UC-MSCs after TGR has the effect of relieving pain and promoting the repair of bone defects, making the operative limb recover function earlier, making up for the deficiency of TGR, and improving the effect of PL treatment. Future studies should furthermore explore the dose and frequency of therapy based on the multiple advantages of UC-MSCs and the mechanism of cartilage repair in dogs.

Analysis of the Role and Mechanism of ZEB1 in Regulating Cervical Carcinoma Progression via Modulating PD-1/PD-L1 Checkpoint.

Background: Cervical carcinoma (CC) is a common and highly malignant tumor in women. The involvement of zinc finger E-box binding homeobox 1 (ZEB1) in many kinds of tumors has been well-documented; however, its role and mechanism in CC remain to be clarified. Objective: This study investigated the mechanism of ZEB1 in modulating the growth and metastasis of CC cells. Methods: The expression of ZEB1 in CC tissues and adjacent normal counterparts was determined by reverse transcription-polymerase chain reaction (RT-PCR). The correlation between ZEB1 and patient clinicopathological indexes was analyzed. In vitro, gain and loss functions of ZEB1 were performed in C-33A and HeLa cell lines. The proliferation, migration, and invasion of CC cells were detected by Cell Counting Kit-8 (CCK-8) assay and transwell assay, respectively. The expression levels of apoptosis-related proteins such as BCL2-associated X (Bax), B-cell lymphoma-2 (Bcl2), and Caspase-3, as well as epithelial-mesenchymal transition (EMT)-associated proteins including E-cadherin, Vimentin, and Snail, were measured by Western blotting. In addition, the targeting relationship between ZEB1 and programmed death 1 (PD-1)/programmed cell death-ligand 1 (PD-L1) was predicted by bioinformatics and further verified by dual-luciferase reporter assay. Results: ZEB1 was significantly up-regulated in CC tissues compared with normal counterparts. ZEB1 overexpression promoted the migration, proliferation, and invasion of CC cells and inhibited apoptosis, while knocking down ZEB1 contributed to the opposite effects. In addition, experiments on related mechanisms confirmed that ZEB1 targeted the 3'EUTR terminal of PD-1/PD-L1 and negatively regulated its expression. And an interaction between ZEB1 and PD-1/PD-L1 was identified. Conclusion: ZEB1 can promote the proliferation and metastasis of CC cells via modulating the PD-1/PD-L1 checkpoint.

Effects of Medical Masks on Voice Quality in Patients With Voice Disorders.

PURPOSE: The purpose of this study was to explore the effects of medical masks on the voice quality of patients with voice disorders. METHOD: We included 106 patients diagnosed with voice disorders. Among them, 59 were diagnosed with vocal-fold benign lesions, 27 with insufficient glottis closure, and 20 with precancerous lesions/early-stage glottic carcinoma. Perceptual parameters (GRBAS [grade, roughness, breathiness, asthenia, strain] scale), acoustic parameters (f o, sound pressure level [SPL], jitter, shimmer, noise-to-harmonic ratio [NHR], and cepstral peak prominence [CPP]), and maximum phonation time (MPT) without and with medical masks were analyzed. Changes in the GRBAS scale after wearing medical masks were also evaluated. RESULTS: With medical mask wearing, the G, R, and B scales in the vocal-fold benign lesion and insufficient glottic closure groups decreased, with a statistical significance seen in the G and R scales of the vocal-fold benign lesion group (G 1.07 ± 0.59, 0.95 ± 0.68, p < .01; R 1.07 ± 0.59, 0.95 ± 0.68, p < .01). The B scale in the precancerous lesions/early-stage glottic carcinoma (95%) and vocal-fold benign lesion groups (83%) and R scale in the insufficient glottic closure group (77.8%) were stable with mask wearing. f o and SPL in the vocal-fold benign lesion group and f o and jitter in the insufficient glottic closure group increased significantly with medical masks. The NHR and CPP in each group changed little, and all the parameters in the precancerous lesions/early-stage glottic carcinoma group showed no significant change. CONCLUSIONS: The effects of medical masks on the voice quality of patients with voice disorders were associated with the type of the disease, degree of hoarseness, and subjective scale influencing specific voice disorder. When wearing medical masks, the pitch and loudness of patients increased as compensation. Medical masks had the least impact on the precancerous lesions/early-stage glottic carcinoma group.

Activatable Nanoprobe with Aggregation-Induced Dual Fluorescence and Photoacoustic Signal Enhancement for Tumor Precision Imaging and Radiotherapy.

Owing to the high sensitivity and high spatial resolution, fluorescence (FL) imaging has been widely applied for visualizing biological processes. To gain insight into molecular events on deeper tissues, photoacoustic (PA) imaging with better deep-tissue imaging capability can be incorporated to provide complementary visualization and quantitative information on the pathological status. However, the development of activatable imaging probes to achieve both FL and PA signal amplification remains challenging because the enhancement of light absorption in PA imaging often caused the quenching of FL signal. Herein, we first developed a caspase-3 enzyme activatable nanoprobe of a nanogapped gold nanoparticle coated with AIE molecule INT20 and DEVD peptides (AuNNP@DEVD-INT20) for tumor FL and PA imaging and subsequent imaging-guided radiotherapy. The nanoprobe could interact with GSH and caspase-3 enzyme to liberate INT20 molecules, leading to AIE. Simultaneously, the in situ self-assembly of AuNPs was achieved through the cross-linking reaction between the sulfhydryl and the maleimide, resulting in ratiometric PA imaging in tumor. Remarkably, the nanoprobe can generate richful ROS for cancer radiotherapy under X-ray irradiation. The platform not only achieves the aggregation-induced FL and PA signal enhancement but also provides a general strategy for imaging of various biomarkers, eventually benefiting precise cancer therapy.

An aptamer-tethered DNA origami amplifier for sensitive and accurate imaging of intracellular microRNA.

Accurate detection and imaging of low-abundance microRNA (miRNA) in living cells are essential for the diagnosis and prognosis of diseases. Designing nanoprobes with resistance to enzyme degradation, effective cell-binding, and efficient signal amplification is crucial for in vivo imaging. In this study, we present an aptamer-tethered DNA origami amplifier (ADOA) that functions inside living cells to detect miRNA with high sensitivity and stability. In the design, cancer cell-targeting aptamers were tethered onto the border of the DNA origami to improve the discrimination between cancer cells and normal cells. Two substrate modules for the intramolecular entropy-driven reaction (EDR) circuit were alternately arranged on the DNA origami plane. The target miRNA will initiate the sequential hybridization of the two substrate modules on the DNA origami, generating amplified fluorescence signals. The proposed ADOA achieved an accelerated cascade reaction due to the "confinement effect" and significantly enhanced the sensitivity compared with a traditional EDR. Meanwhile, with the rigid structure of the DNA origami, the ADOA possessed excellent signalling stability in living cells. Therefore, the ADOA could expand the application of DNA origami in miRNA sensing and has potential value in early-stage clinical diagnosis.

Multifunctional Carbon Monoxide Prodrug-Loaded Nanoplatforms for Effective Photoacoustic Imaging-Guided Photothermal/Gas Synergistic Therapy.

Multifunctional cancer treatments based on gas therapy combined with other cancer treatments have gained tremendous attention and hold great promise in biomedical applications. In this study, a carbon monoxide-releasing nanoplatform combined with near-infrared (NIR) laser-triggered photothermal therapy (PTT) was constructed. The nanoplatform was composed of manganese pentacarbonyl bromide (MnCO)-loaded g-carbon nitride/polypyrrole (CNPpy) nanomaterials (MnCO@CNPpy). MnCO can be triggered to produce CO under H2O2 conditions. Upon exogenous NIR light stimulation and tumor microenvironment-overexpressed H2O2, MnCO@CNPpy exhibited excellent CO generation performance and photothermal effect. The generation of CO induced intracellular oxidative stress and caused cell apoptosis. Additionally, photoacoustic (PA) imaging was performed to track the delivery and accumulation of the nanomaterial in tumor sites because of the great photothermal conversion of CNPpy. The presented MnCO@CNPpy nanoplatform displayed desirable PTT and CO therapy in the inhibition of tumor growth and may provide a promising strategy for multifunctional antitumor synergistic treatments.

Functional Self-Assembled DNA Nanohydrogels for Specific Telomerase Activity Imaging and Telomerase-Activated Antitumor Gene Therapy.

Engineering a functional nanoplatform that integrates dynamic monitoring of endogenous biomarkers and a stimuli-activated therapeutic mode is promising for early diagnosis and treatment of cancers. In this study, we developed an intelligent DNA nanohydrogel with specific targeting capability that can be stimuli-activated for both in vitro telomerase detection and in vivo telomerase-triggered gene therapy. The DNA nanohydrogel was formed simply by the self-assembly of two Y-shaped DNA units and a double-stranded DNA linker labeled with fluorophores and loaded with therapeutic siRNA. When intracellular telomerase was overexpressed, the DNA nanohydrogel collapsed owing to the prolongation of the telomeric primer at the terminal sequence of one of the Y-shaped DNA units. As a result, the quenched fluorescence due to fluorescence resonance energy transfer (FRET) of the DNA nanohydrogel recovered and the trapped siRNA was released, enabling the accurate detection and imaging of intracellular telomerase activity as well as effective gene therapy of tumors. Benefiting from the great biocompatibility, specificity, and stimuli-responsive property, the developed DNA nanoplatform provides a new opportunity for precise cancer diagnosis and treatment as well as other biological applications.

An electrochemical sensor based on enzyme-free recycling amplification for sensitive and specific detection of miRNAs from cancer cells.

MicroRNAs (miRNAs) are desirable targets for the diagnosis, prognosis and treatment of diverse human diseases. In this study, we developed a universal and enzyme-free signal amplification approach for the quick, sensitive and specific electrochemical detection of diverse miRNAs from tumor cells using a catalyzed hairpin assembly (CHA) and the DNA three-way junction (DNA TWJ). The target miRNA, which was formed as an initiator through specific recognition, periodically triggered the assembly of two hairpins into a duplex via CHA. Subsequently, these duplexes induced the catalytic assembly of the DNA three-way junction and the release of the methylene blue-labeled DNA strand S (S-MB). Thereafter, the S-MB was captured using the electrode through hybridization with the capture probe to generate a notable current response. The proposed biosensor could be switched in response to different miRNA targets by changing the specific sequence of H0. These findings indicate its ability to distinguish single base mismatch miRNAs and analyze miRNAs extracted from cells. Therefore, the universal and enzyme-free signal amplified electrochemical method described herein can be potentially useful in diverse miRNA-related clinical analysis and disease diagnosis.

Transcriptome profile of rat genes in bone marrow-derived macrophages at different activation statuses by RNA-sequencing.

The underlying mechanisms of macrophage polarization have been detected by genome-wide transcriptome analysis in a variety of mammals. However, the transcriptome profile of rat genes in bone marrow-derived macrophages (BMM) at different activation statuses has not been reported. Therefore, we performed RNA-Sequencing to identify gene expression signatures of rat BMM polarized in vitro with different stimuli. The differentially expressed genes (DEGs) among unactivated (M0), classically activated pro-inflammatory (M1), and alternatively activated anti-inflammatory macrophages (M2) were analyzed by using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis. In this study, not only we have identified the changes of global gene expression in rat M0, M1 and M2, but we have also made clear systematically the key genes and signaling pathways in the differentiation process of M0 to M1 and M2. These will provide a foundation for future researches of macrophage polarization.

DNA-mediated reversible capture and release of circulating tumor cells with a multivalent dual-specific aptamer coating network.

DNA-triggered reversible isolation and recovery of circulating tumor cells (CTCs) is presented based on a multivalent dual-specific aptamer-tethered rolling circle amplification (MA-RCA) network. The multivalent binding sites endow the MA-RCA network with a strong binding ability towards CTCs, and the repeated cell capture/release processes are also actualized in a noninvasive manner.