An Activity-Based Ratiometric Fluorescent Probe for In†Vivo Real-Time Imaging of Hydrogen Molecules.

To reveal the biomedical effects and mechanisms of hydrogen molecules urgently needs hydrogen molecular imaging probes as an imperative tool, but the development of these probes is extremely challenging. A catalytic hydrogenation strategy is proposed to design and synthesize a ratiometric fluorescent probe by encapsulating Pd nanoparticles and conjugating azido-/coumarin-modified fluorophore into mesoporous silica nanoparticles, realizing in vitro and in vivo fluorescence imaging of hydrogen molecules. The developed hydrogen probe exhibits high sensitivity, rapid responsivity, high selectivity and low detection limit, enabling rapid and real-time detection of hydrogen molecules both in cells and in the body of animal and plant. By application of the developed fluorescent probe, we have directly observed the super-high transmembrane and ultrafast transport abilities of hydrogen molecules in cells, animals and plants, and discovered in vivo high diffusion of hydrogen molecules.

Sulourea-coordinated Pd nanocubes for NIR-responsive photothermal/H2S therapy of cancer.

BACKGROUND: Photothermal therapy (PTT) frequently cause thermal resistance in tumor cells by inducing the heat shock response, limiting its therapeutic effect. Hydrogen sulfide (H2S) with appropriate concentration can reverse the Warburg effect in cancer cells. The combination of PTT with H2S gas therapy is expected to achieve synergistic tumor treatment. METHODS: Here, sulourea (Su) is developed as a thermosensitive/hydrolysable H2S donor to be loaded into Pd nanocubes through in-depth coordination for construction of the Pd-Su nanomedicine for the first time to achieve photo-controlled H2S release, realizing the effective combination of photothermal therapy and H2S gas therapy. RESULTS: The Pd-Su nanomedicine shows a high Su loading capacity (85 mg g-1), a high near-infrared (NIR) photothermal conversion efficiency (69.4%), and NIR-controlled H2S release by the photothermal-triggered hydrolysis of Su. The combination of photothermal heating and H2S produces a strong synergetic effect by H2S-induced inhibition of heat shock response, thereby effectively inhibiting tumor growth. Moreover, high intratumoral accumulation of the Pd-Su nanomedicine after intravenous injection also enables photothermal/photoacoustic dual-mode imaging-guided tumor treatment. CONCLUSIONS: The proposed NIR-responsive heat/H2S release strategy provides a new approach for effective cancer therapy.

Nanoliposomes Co-Encapsulating Photoswitchable Probe and Photosensitizer for Super-Resolution Optical Imaging and Photodynamic Therapy.

Photosensitizers (PSs) are ideal cancer theranostic drugs that can be administered as both fluorescence imaging reagents and photodynamic therapy (PDT) drugs. To improve the tumoritropic behavior of PSs, nanoliposomes are presently being considered as optimal PSs carriers. Although nanoliposomal PSs have been utilized in clinical therapy, PSs localization and photosensitive processing in nanoliposomal PSs are rarely observed on nanoscale. Investigating changes in the fine structure of nanoliposomes under photosensitive processing will further our understanding of the photosensitive effect on nanoliposomal PSs. In this study, nanoliposomes co-encapsulating the PSs benzoporphyrin derivative monoacid A (BPD) and the photoswitchable probe Cy5-927 were prepared to realize PDT and nanoscale super-resolution optical imaging. The fine structures of nanoliposomal BPD and Cy5-927 (LBC) were visualized by a home-built stochastic optical reconstruction microscopy (STORM). Our PDT results showed that the photorelease and PDT efficiency of BPD were not decreased by co-encapsulating with Cy5-927 in LBC. Taken together, LBC can be used as a new optical probe and PDT reagent for investigating changes in nanoliposomes fine structure and micro-interaction in the cellular process of PDT. Therefore, our results deepened our understanding of liposome-based PDT for optimizing cancer treatment. © 2019 International Society for Advancement of Cytometry.

Inhibition and Stabilization: Cucurbituril Induced Distinct Effects on the Schiff Base Reaction.

The different effects of cucurbit[7]uril (CB[7]) on the Schiff base reactions in aqueous solution were explored by 1H NMR spectroscopy and single X-ray crystallography. With CB[7], the condensation reaction of aldehyde and primary amine is dramatically inhibited. In contrast, the presence of CB[7] does tremendously stabilize iminium cation in water through ion-dipole interactions. A single crystal structure of the complex of iminium ion 7 with CB[7] grown in water is reported.

From Packed "Sandwich" to "Russian Doll": Assembly by Charge-Transfer Interactions in Cucurbit[10]uril.

As the host possessing the largest cavity in the cucurbit[n]uril (CB[n]) family, CB[10] has previously displayed unusual recognition and assembly properties with guests but much remains to be explored. Herein, we present the recognition properties of CB[10] toward a series of bipyridinium guests including the tetracationic cyclophane known as blue box along with electron-rich guests and detail the influence of encapsulation on the charge-transfer interactions between guests. For the mono-bipyridinium guest (methylviologen, MV2+ ), CB[10] not only forms 1:1 and 1:2 inclusion complexes, but also enhances the charge-transfer interactions between methylviologen and dihydroxynaphthalene (HN) by mainly forming the 1:2:1 packed "sandwich" complex (CB[10]⋅2 MV2+ ⋅HN). For guest 1 with two bipyridinium units, an interesting conformational switching from linear to "U" shape is observed by adding catechol to the solution of CB[10] and the guest. For the tetracationic cyclophane-blue box, CB[10] forms a stable 1:1 inclusion complex; the two bipyridinium units tilt inside the cavity of CB[10] according to the X-ray crystal structure. Finally, a supramolecular "Russian doll" was built up by threading a guest through the cavities of both blue box and CB[10].

Therapeutic gas delivery strategies.

Gas molecules with pharmaceutical effects offer emerging solutions to diseases. In addition to traditional medical gases including O2 and NO, more gases such as H2 , H2 S, SO2 , and CO have recently been discovered to play important roles in various diseases. Though some issues need to be addressed before clinical application, the increasing attention to gas therapy clearly indicates the potentials of these gases for disease treatment. The most important and difficult part of developing gas therapy systems is to transport gas molecules of high diffusibility and penetrability to interesting targets. Given the particular importance of gas molecule delivery for gas therapy, distinguished strategies have been explored to improve gas delivery efficiency and controllable gas release. Here, we summarize the strategies of therapeutic gas delivery for gas therapy, including direct gas molecule delivery by chemical and physical absorption, inorganic/organic/hybrid gas prodrugs, and natural/artificial/hybrid catalyst delivery for gas generation. The advantages and shortcomings of these gas delivery strategies are analyzed. On this basis, intelligent gas delivery strategies and catalysts use in future gas therapy are discussed. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.

DNA damage response in resting and proliferating peripheral blood lymphocytes treated by camptothecin or X-ray.

DNA damage response (DDR) in different cell cycle status of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA). The apoptotic ratio and the phosphorylation H2AX (S139) were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P<0.05). The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.

Super-resolution imaging of the dynamic cleavage of intercellular tunneling nanotubes.

As a new method of cell-cell communication, tunneling nanotubes (TNTs) play important roles in cell-cell signaling and mass exchanges. However, a lack of powerful tools to visualize dynamic TNTs with high temporal/spatial resolution restricts the exploration of their formation and cleavage, hindering the complete understanding of its mechanism. Herein, we present the first example of using stochastic optical reconstruction microscopy (STORM) to observe the tube-like structures of TNTs linking live cells with an easily prepared fluorescent dye. Because of this new imaging microscopy, the cleavage process of TNTs was observed with a high spatial resolution.

STORM imaging of mitochondrial dynamics using a vicinal-dithiol-proteins-targeted probe.

Stochastic optical reconstruction microscopy (STORM) is a promising method for the visualization of ultra-fine mitochondrial structures. However, this approach is limited to monitoring dynamic intracellular events owing to its low temporal resolution. We developed a new strategy to capture mitochondrial dynamics using a compressed sensing STORM algorithm following raw data pre-treatments by a noise-corrected principal component analysis and K-factor image factorization. Using STORM microscopy with a vicinal-dithiol-proteins targeting probe, visualizing mitochondrial dynamics was attainable with spatial and temporal resolutions of 45 nm and 0.8 s, notably, dynamic mitochondrial tubulation retraction of ~746 nm in 1.2 s was monitored. The labeled conjugate was observed as clusters (radii, ~90 nm) distributed on the outer mitochondrial membranes, not yet reported as far as we know. This strategy is promising for the quantitative analysis of intracellular behaviors below the optical diffraction limit.

Long non-coding RNA SNHG1 indicates poor prognosis and facilitates disease progression in acute myeloid leukemia.

The role of long non-coding RNAs (lncRNAs) in acute myeloid leukemia (AML) is becoming increasingly questioned. Previous studies have reported that the lncRNA small nucleolar RNA host gene 1 (SNHG1) is involved in multiple human malignant tumors, while its expression and role in AML is still unexplored. Here, we show that SNHG1 is highly expressed in AML specimens from non-M3 patients, as well as AML cell lines. Meanwhile, upregulation of SNHG1 is correlated with poor prognosis. Notably, SNHG1 facilitates the proliferation and inhibits the apoptosis of AML cells in vitro Consistent with these findings, knockdown of SNHG1 significantly inhibits AML progression in an immunodeficient mouse model. Mechanistically, we found that an anti-tumor microRNA-101 (miR-101) is upregulated and its target genes are downregulated in AML cells after SNHG1 knockdown. Further investigations display that SNHG1 can serve as a competing endogenous RNA to inhibit miR-101. In conclusion, our data indicate that SNHG1 plays an important role in facilitating AML progression at least in part by negatively regulating miR-101, and provides a new target for treating AML.

Redefining the photo-stability of common fluorophores with triplet state quenchers: mechanistic insights and recent updates.

Light microscopy can offer certain advantages over electron microscopy in terms of acquiring detailed insights into the biological/intra-cellular milieu. In recent years, with the development of new fluorescence imaging technologies, it has become extremely important to assess the role of designing appropriate fluorophores in acquiring desired biological information without encountering any untoward hitches. Over the years, external fluorophores have been prevalently used in fluorescence microscopy and single-molecule fluorescence microscopy-based studies. Photostable fluorogenic probes with high extinction coefficients and quantum yields, exhibiting minimum autofluorescence and photobleaching properties, are preferred in single-molecule microscopy as they can tolerate long-term laser exposure. Therefore, the development of triplet state quenchers and/or any other suitable new strategy to ensure the photo-stability of the fluorophores during long-term live cell imaging exercises is highly anticipated. In this feature article, various strategies for stabilizing fluorophores, including the mechanisms of TSQ-induced stabilization, have been thoroughly reviewed considering contemporary literature reports and applications.

Dual-functional fluorescent molecular rotor for endoplasmic reticulum microviscosity imaging during reticulophagy.

The microviscosity change associated with reticulophagy is an important component for studying endoplasmic reticulum (ER) stress disorders. Here, a BODIPY-arsenicate conjugate 1-based fluorescent molecular rotor was designed to covalently bind vicinal dithiol-containing proteins in the ER, exhibiting a bifunction of reticulophagy initiation and microviscosity evaluation. Therefore, we could quantify the local viscosity changes during reticulophagy based on the fluorescence lifetime changes of probe 1.

Elevated serum level of lncRNA-HIF1A-AS1 as a novel diagnostic predictor for worse prognosis in colorectal carcinoma.

To evaluate the diagnostic efficacy and prognostic value of serum long non-coding RNA (lncRNA) HIF 1alpha-antisense RNA 1 (HIF1A-AS1) in patients with colorectal carcinoma (CRC). We obtained serum samples from 151 CRC patients and 160 health controls. Serum level of HIF1A-AS1 was detected via real-time PCR (RT-PCR). Receiver operating characteristics (ROC) curve analysis was conducted to determine the diagnostic value of HIF1A-AS1. Then HIF1A-AS1 in CRC was divided into high- and low-expression groups, and the associations of the HIF1A-AS1 serum level with clinicopathological features and prognosis were analyzed. Serum level of HIF1A-AS1 was significantly increased from CRC patients as compared to those of health controls (P< 0.05). ROC curve analysis revealed a relative high diagnostic performance of HIF1A-AS1 to distinguish CRC from health controls, with the area under the curves (AUC) of 0.960 (95% CI: 0.940 ∼ 0.980; P< 0.001). Kaplan-Meier analysis showed that differentiation degree, tumor size, TNM stage, T stage, N stage, M stage and serum level of HIF1A-AS1 were all linked to CRC prognosis (All P< 0.05). Compared to CRC patients with low HIF1A-AS1 expression, high expression of patients were associated with a shorter 5-year-survival rate (P< 0.001). Multivariate Cox regression analysis revealed that lower differentiation degree, tumor > 5 cm and higher expression of HIF1A-AS1 were independent risk factors affecting the survival rate of patients with CRC (P< 0.05). Our results illustrated that elevated serum HIF1AAS1 could be clinically functioned as a potential biomarker for CRC diagnoses and prognosis.

DNA Damage Response in Resting and Proliferating Peripheral Blood Lymphocytes Treated by Camptothecin or X-ray / 华中科技大学学报（医学）（英德文版）

DNA damage response (DDR) in different cell cycle starus of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated.The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA).The apoptotic ratio and the phosphorylation H2AX (S139)were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray.The expressions of γH2AX,Bcl-2,caspase-3 and caspase-9 were detected by Western blotting.DDR in 293T cells was detected after H2AX was silenced by RNAi method.Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment.The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P＜0.05).The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased.No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls.It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates.γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.