Dapagliflozin Improves Angiogenesis after Hindlimb Ischemia through the PI3K-Akt-eNOS Pathway.

Recently, the vascular protective effect of anti-diabetic agents has been receiving much attention. Sodium glucose cotransporter 2 (SGLT2) inhibitors had demonstrated reductions in cardiovascular (CV) events. However, the therapeutic effect of dapagliflozin on angiogenesis in peripheral arterial disease was unclear. This study aimed to explore the effect and mechanism of dapagliflozin on angiogenesis after hindlimb ischemia. We first evaluated the effect of dapagliflozin on post-ischemic angiogenesis in the hindlimbs of rats. Laser doppler imaging was used to detect the hindlimb blood perfusion. In addition, we used immunohistochemistry to detect the density of new capillaries after ischemia. The relevant signaling pathways of dapagliflozin affecting post-ischemic angiogenesis were screened through phosphoproteomic detection, and then the mechanism of dapagliflozin affecting post-ischemic angiogenesis was verified at the level of human umbilical vein endothelial cells (HUVECs). After subjection to excision of the left femoral artery, all rats were randomly distributed into two groups: the dapagliflozin group (left femoral artery resection, receiving intragastric feeding with dapagliflozin (1 mg/kg/d), for 21 consecutive days) and the model group, that is, the positive control group (left femoral artery resection, receiving intragastric feeding with citric acid-sodium citrate buffer solution (1 mg/kg/d), for 21 consecutive days). In addition, the control group, that is the negative control group (without left femoral artery resection, receiving intragastric feeding with citric acid-sodium citrate buffer solution (1 mg/kg/d), for 21 consecutive days) was added. At day 21 post-surgery, the dapagliflozin-treatment group had the greatest blood perfusion, accompanied by elevated capillary density. The results showed that dapagliflozin could promote angiogenesis after hindlimb ischemia. Then, the ischemic hindlimb adductor-muscle tissue samples from three rats of model group and dapagliflozin group were taken for phosphoproteomic testing. The results showed that the PI3K-Akt-eNOS signaling pathway was closely related to the effect of dapagliflozin on post-ischemic angiogenesis. Our study intended to verify this mechanism from the perspective of endothelial cells. In vitro, dapagliflozin enhanced the tube formation, migration, and proliferation of HUVECs under ischemic and hypoxic conditions. Additionally, the dapagliflozin administration upregulated the expression of angiogenic factors phosphorylated Akt (p-Akt) and phosphorylated endothelial nitric oxide synthase (p-eNOS), as well as vascular endothelial growth factor A (VEGFA), both in vivo and in vitro. These benefits could be blocked by either phosphoinositide 3-kinase (PI3K) or eNOS inhibitor. dapagliflozin could promote angiogenesis after ischemia. This effect might be achieved by promoting the activation of the PI3K-Akt-eNOS signaling pathway. This study provided a new perspective, new ideas, and a theoretical basis for the treatment of peripheral arterial disease.

Identification of acute myeloid leukemia by infrared difference spectrum of peripheral blood.

Acute myeloid leukemia (AML) is a high mortality and recurrence rates hematologic malignancy. Thus, whatever early detection or subsequent visit are both of high significance. Traditional AML diagnosis is conducted via peripheral blood (PB) smear and bone marrow (BM) aspiration. But BM aspiration is a painful burden for patients especially in early detection or subsequent visit. Herein, the use of PB to evaluate and identify the leukemia characteristics will be an attractive alternative source for early detection or subsequent visit. Fourier transform infrared spectroscopy (FTIR) is a time- and cost-effective approach to reveal the disease-related molecular features and variations. However, to the best of our knowledge, there is no attempts using infrared spectroscopic signatures of PB to replace BM for identifying AML. In this work, we are the first to develop a rapid and minimally invasive method to identify AML by infrared difference spectrum (IDS) of PB with only 6 characteristic wavenumbers. We dissect the leukemia-related spectroscopic signatures of three subtypes of leukemia cells (U937, HL-60, THP-1) by IDS, revealing biochemical molecular information about leukemia for the first time. Furthermore, the novel study links cellular features to complex features of blood system which demonstrates the sensitivity and specificity with IDS method. On this basis, BM and PB of AML patients and healthy controls were provided to parallel comparison. The IDS of BM and PB combined with principal component analysis method revealing that the leukemic components in BM and PB can be described by IDS peaks of PCA loadings, respectively. It is demonstrated that the leukemic IDS signatures of BM can be replaced by the leukemic IDS signatures of PB. In addition, the IDS signatures of leukemia cells are reflected in PB of AML patients with peaks of 1629, 1610, 1604, 1536, 1528 and 1404 cm-1 for the first time as well. To this end, we access the leukemic signatures of IDS peaks to compare the PB of AMLs and healthy controls. It is confirmed that the leukemic components can be detected from PB of AML and distinguished into positive (100%) and negative (100%) groups successfully by IDS classifier which is a novel and unique spectral classifier. This work demonstrates the potential use of IDS as a powerful tool to detect leukemia via PB which can release subjects' pain remarkably.

Erratum: Comparative study of nanostructured carriers of calcium phosphate and magnesium phosphate loaded with SRT1720 for the protection of H2O2-induced senescent endothelium.

[This corrects the article on p. 2068 in vol. 10, PMID: 30093944.].

Robotic written silver ink on photographic paper for detection of thiram residues in fruits.

With the widespread application of pesticide in agriculture, pesticide residues in food have posed serious health risks to human. There is an urgent requirement to detect pesticide residues in food. In this work, a sensitive and effective method was employed to measure thiram residues in fruit using surface-enhanced Raman scattering (SERS) technique. Silver ink was written on photographic paper (AgNPs-photographic paper) directly by robotic writing technique. The AgNPs-photographic paper substrates possessed good SERS activities and high stability among four months. A good linear response between the peaks intensities and the logarithmic concentrations of thiram was obtained with the limit of detection (LOD) of 0.024 ppb. The substrates also exhibited excellent reproducibility with relative standard deviation (RSD) value less than 10% from ten different substrates. SERS mapping was tested to characterize the uniformity of AgNPs-photographic paper, and the RSD value was calculated to be 14.34% at 1377 cm-1 measured by 120 points. The LOD values of apple and peach juice adulterated with thiram were 0.0024 and 0.024 ppm, respectively. The LOD values of thiram residues on apple and peach peels were both 0.25 ng/cm2. It was demonstrated that the substrates prepared by robotic writing technique had great potential for practical application in food safety inspection.

Correction: Highly porous and elastic aerogel based on ultralong hydroxyapatite nanowires for high-performance bone regeneration and neovascularization.

Correction for 'Highly porous and elastic aerogel based on ultralong hydroxyapatite nanowires for high-performance bone regeneration and neovascularization' by Gao-Jian Huang et al., J. Mater. Chem. B, 2021, 9, 1277-1287, DOI: 10.1039/D0TB02288H.

Efficient fabrication of highly sensitive AgNPs-drawing paper SERS substrates by robotic writing approach.

Inspired by hand writing approach for preparing surface-enhanced Raman scattering (SERS) substrates, silver nanoparticles (AgNPs) decorated drawing paper substrates were prepared by robotic writing technique. The wettabilities and surface morphologies of the drawing paper before and after the deposition of AgNPs were characterized by contact angle analyzer and scanning electron microscope, respectively. Malachite green was employed as a probe molecule to evaluate the SERS activities of the AgNPs-drawing paper substrates. The AgNPs-drawing paper substrates exhibited extremely high sensitivity that the detection limit for malachite green was down to 10-18 mol/L and the Raman enhancement factor was calculated to be 1015. The relative standard deviation (RSD) values of the Raman peaks intensities collected from twelve points on a single substrate and fifteen substrates were used to evaluate the uniformity and reproducibility of the AgNPs-drawing paper substrates. It was found that the substrates had good reproducibility and uniformity with RSD values of 7.29% and 9.70%, respectively. Furthermore, the prepared AgNPs-drawing paper substrates exhibited long-term stability among six months.

Highly porous and elastic aerogel based on ultralong hydroxyapatite nanowires for high-performance bone regeneration and neovascularization.

Hydroxyapatite (HAP) is promising for the clinical treatment of bone defects because of its excellent biocompatibility and osteo-conductivity. However, highly porous HAP scaffolds usually exhibit high brittleness and poor mechanical properties, thus organic constituents are usually added to form composite materials. In this work, a highly porous and elastic aerogel made from ultralong HAP nanowires with ultrahigh porosity (∼98.5%), excellent elasticity and suitable porous structure is prepared as the high-performance scaffold for bone defect repair. The highly porous structure of the as-prepared aerogel is beneficial to bone ingrowth and matter/fluid transfer, and the high elasticity can ensure the structural integrity of the scaffold during bone regeneration. Therefore, the HAP nanowire aerogel scaffold can promote the adhesion, proliferation and migration of rat bone marrow derived mesenchymal stem cells (rBMSCs), and elevate the protein expression of osteogenesis and angiogenesis related genes. The in vivo experimental results demonstrate that the HAP nanowire aerogel scaffold is favorable for the ingrowth of new bone and blood vessels, and thus can greatly accelerate bone regeneration and neovascularization. The as-prepared HAP nanowire aerogel scaffold shows promising potential for biomedical applications such as bone defect repair.

Graphene Quantum Dots with Pyrrole N and Pyridine N: Superior Reactive Oxygen Species Generation Efficiency for Metal-Free Sonodynamic Tumor Therapy.

Those responsible for the development of sonosensitizers are faced with a dilemma between high sonosensitization efficacy and good biosecurity that limited the development of sonodynamic therapy (SDT). Herein, inspired by the intriguing therapeutic features of SDT and the potential catalytic activity of graphene quantum dots, the potential of N-doped graphene quantum dots (N-GQDs) to act as a sonosensitizer is demonstrated. The superior sonosensitization effect of N-GQDs is believed to be three to five times higher than that of traditional sonosensitizers (such as porphyrin, porphyrin Mn, porphyrin Zn, TiO2 , etc.). More importantly, the sonochemical mechanism of N-GQDs is revealed. Pyrrole N and pyridine N are believed to form catalytic centers in sonochemical processing of N-GQDs. This knowledge is important from the perspective of understanding the structure-dependent SDT enhancement of carbon nanostructure. Moreover, N-GQDs modified by folic acid (FA-N-GQDs) show a high marker rate for tumor cells (greater than 96%). Both in vitro and in vivo therapeutic results have exhibited high tumor inhibition efficiency (greater than 90%) of FA-N-GQDs as sonosensitizers while the oxidative stress response of tumor cells is activated through the PEX pathway and induced apoptosis via the p53 pathway.

Inkjet printed silver nanoparticles on hydrophobic papers for efficient detection of thiram.

Silver nanoparticles coated paper (AgNPs-paper) substrates were prepared by inkjet printing Ag ink on four different wettability papers. Scanning electron microscope and contact angle analyzer were used to characterize their surface morphology and wettability. AgNPs-paper substrates were used to detect the surface-enhanced Raman scattering (SERS) spectra of thiram aqueous solution. Relationships between the surface wettability, surface morphology and SERS activities of the substrates were systematically studied. The silver nanoparticles deposited on the hydrophobic papers (photographic paper, graph paper, and weighing paper) were evenly and densely arranged. While in-homogeneous distribution was observed on the hydrophilic printing paper. It can be found that the AgNPs-photographic paper with the maximum contact angle exhibited the highest SERS enhancement. The detection limit for thiram adsorbed on the AgNPs-photographic paper was 10-10 mol/L, which was lower than the others. Good linear responses (R2 = 0.9918, 0.9897) between the SERS intensities and logarithmic concentrations were obtained from 104 to 10-10 mol/L. Moreover, the substrate had good uniformity and reproducibility with relative standard deviation values of 4.20% and 4.90% measured by eight points and ten substrates, respectively. The AgNPs-photographic paper exhibited high stability within eight months.

Fabrication and SERS performance of silver nanoarrays by inkjet printing silver nanoparticles ink on the gratings of compact disc recordable.

Silver nanoarrays were fabricated by inkjet printing silver nanoparticles ink on the gratings of compact disc recordable (CD-R). Rhodamine 6G (R6G) was chosen as a probe molecule to evaluate their surface-enhanced Raman scattering (SERS) performance. The finite-difference time domain (FDTD) solution was used to simulate local electric field distribution of silver nanoparticles on the grating surface and flat surface, respectively. It was found that the Ag/grating substrate possessed higher enhancement ability than the Ag/flat due to the high-density hot spots of periodic structure of the grating. The silver nanoarrays substrate exhibited high stability and the characteristic peaks of R6G can be still well observed after eight months. The substrate also exhibited a good spot-to-spot reproducibility with an RSD of 10.21% by eight points. SERS mappings of R6G adsorbed on silver nanoarrays were tested under the ultra-fast Raman imaging mode, and the relative standard deviation (RSD) values of uniformity were calculated to be 8.35% and 11.53% at 610 cm-1 band measured by 2500 and 6480 points, respectively. In addition, the as-prepared silver nanoarrays was successfully applied to the detection of melamine in adult milk powder solution directly. A good linear relationship with the correlation coefficient of 0.9968 between peak intensity and concentration was obtained from 1.2 to 100 mg/L.

Atorvastatin plus therapeutic ultrasound improve postnatal neovascularization in response to hindlimb ischemia via the PI3K-Akt pathway.

Statins and therapeutic ultrasound (TUS) have been shown to ameliorate angiogenesis on ischemic hindlimb animals and promote human umbilical vein endothelial cells (HUVECs) tube formation and proliferation. Here, we evaluate the therapeutic effect of TUS in combination with atorvastatin (Ator) therapy on angiogenesis in hindlimb ischemia and HUVECs. After subjecting excision of the left femoral artery, all mice were randomly distributed to one of four groups: Control; Ator treated mice (Ator); TUS treated mice (TUS); and Ator plus TUS treated mice (Ator+TUS). At day 14 post-surgery, the Ator plus TUS treatment cohort had the greatest blood perfusion, accompanied by elevated capillary density. In vitro, Ator plus TUS augmented tube formation, migration and proliferative capacities of HUVECs. Additionally, the united administration upregulated expression of angiogenic factors phosphorylated Akt (p-Akt), phosphorylated endothelial nitric oxide synthase (p-eNOS), as well as vascular endothelial growth factor (VEGF), both in vivo and in vitro. These benefits could be blocked by either phosphoinositide 3-kinase (PI3K) or eNOS inhibitor. Our data indicated that the united administration could significantly enhance ischemia-mediated angiogenesis and exert a protective effect against ischemic/hypoxia induced damage among HUVECs through up-regulating VEGF expression and activating the PI3K-Akt-eNOS pathway.

Malignant pleural effusion supernatant is an alternative liquid biopsy specimen for comprehensive mutational profiling.

BACKGROUND: The clinical utility of malignant pleural effusion (MPE) to detect mutation has been well documented; however, routine practice of the use of MPE involves collection of the cell pellet to detect mutation, and limited studies have interrogated the MPE supernatant as an alternative source of tumor-derived DNA for mutation profiling. In this study, we investigated the potential of MPE supernatant as a liquid biopsy specimen by comparing its mutation profile with that of matched MPE cell pellets, tissue, and plasma samples. METHODS: Sequencing data from 17 patients with matched lung tissue, plasma, and MPE samples were retrospectively analyzed. Capture-based targeted sequencing was performed on matched plasma and MPE supernatant samples obtained from 154 patients with advanced lung adenocarcinoma. RESULTS: MPE supernatants had significantly higher median maximum allelic fractions (maxAFs) than their corresponding cell pellets (P = 0.008) and plasma samples (P = 0.036), and a comparable maxAF value to that of tissue samples (P = 0.675). Comparison of MPE supernatant and matched plasma samples from the larger cohort (n = 154) revealed a comparable mutation detection rate; however, MPE supernatant had a significantly higher median maxAF than plasma (20.3% vs. 1.13%; P < 0.001). Furthermore, the concordance rates between MPE supernatant and plasma for single-nucleotide and copy number variations were 56% and 18%, respectively, suggesting that MPE supernatant reveals a more comprehensive mutation spectrum, particularly for copy number variations. CONCLUSION: Overall, our study shows that MPE supernatant is an optimal alternative source of tumor-derived DNA for comprehensive mutation profiling.

Therapeutic ultrasound potentiates the anti-nociceptive and anti-inflammatory effects of curcumin to postoperative pain via Sirt1/NF-κB signaling pathway.

Background: Postoperative pain has well defined and is perceived by patients as one of the most obnoxious aspects of surgical pain. The aim of this study was to determine whether the combination of Therapeutic ultrasound (TUS) and Curcumin (CUR) resulted in an enhancement of their pain relieving activities in a rat model of postoperative pain. Methods: We explored the effect of these treatment and their interaction with signal transduction pathways involved in inflammatory. In this study, TUS and CUR alone or in combination were administered prior to or simultaneously with or after the incisional surgery. Results: At the start time of administration, we observed that the TUS plus CUR treatment reduced the mean paw withdrawal threshold more efficiently than CUR alone. Then we demonstrated that TUS potentiates the antinociceptive effect of CUR in a rat model of chronic postoperative pain and that the combination could facilitate the recovery of surgical pain. However, preventive value was not statistically significant when the treatments were given prior to the incisional surgery. We provide evidence that TUS plus CUR administrations were safe and significantly reduced the ED50 compared to treatment with the single CUR treatment in rats. TUS plus CUR administrations decreases incisional surgery induced activation of inflammatory cells and down-regulation of chemokines and proinflammatory cytokines, MCP-1, MIP-1α, IL-1ß, and TNF-α through regulating Sirt1/NF-κB signaling pathway. Conclusions: Taken together, our results indicate that the combinations of TUS and CUR can be more effective in the anti-nociceptive effects than the treatment with CUR alone.

Comparative study of nanostructured carriers of calcium phosphate and magnesium phosphate loaded with SRT1720 for the protection of H2O2-induced senescent endothelium.

Nanostructured calcium phosphate (CaP) and magnesium phosphate (MgP) are promising for the application as the nanocarriers in drug delivery. However, the difference between CaP and MgP nanocarriers in drug delivery is rarely investigated. In this work, we comparatively investigated nanostructured CaP, MgP and calcium magnesium phosphate (CMP) for the delivery of SRT1720, which is a silent information regulator (SIRT1) specific activator with pro-angiogenic and anti-aging properties in response to hydrogen peroxide (H2O2)-induced endothelial senescence. The protection of SRT1720-loaded CaP nanospheres, MgP nanosheets and CMP microspheres on the H2O2-induced senescent endothelium was examined by using human umbilical vein endothelial cells (HUVECs), demonstrating the improved cell viability, anti-aging, tube formation and migration. In addition, the SRT1720-loaded CaP nanospheres, MgP nanosheets and CMP microspheres can rescue the impaired angiogenic potential of HUVECs via activation of Akt/eNOS/VEGF pathway. The SRT1720-loaded MgP nanosheets and CMP microspheres have a similar protective effect compared with the pure SRT1720, while the SRT1720-loaded CaP nanospheres decrease the protective capability of SRT1720. These results lead us to figure out both MgP nanosheets and CMP microspheres are suitable and effective delivery for SRT1720 and this system can be further applied in vivo treatment.

MiR-27b Promotes Muscle Development by Inhibiting MDFI Expression.

BACKGROUND/AIMS: Skeletal muscle plays an essential role in the body movement. However, injuries to the skeletal muscle are common. Lifelong maintenance of skeletal muscle function largely depends on preserving the regenerative capacity of muscle. Muscle satellite cells proliferation, differentiation, and myoblast fusion play an important role in muscle regeneration after injury. Therefore, understanding of the mechanisms associated with muscle development during muscle regeneration is essential for devising the alternative treatments for muscle injury in the future. METHODS: Edu staining, qRT-PCR and western blot were used to evaluate the miR-27b effects on pig muscle satellite cells (PSCs) proliferation and differentiation in vitro. Then, we used bioinformatics analysis and dual-luciferase reporter assay to predict and confirm the miR-27b target gene. Finally, we elucidate the target gene function on muscle development in vitro and in vivo through Edu staining, qRT-PCR, western blot, H&E staining and morphological observation. RESULT: miR-27b inhibits PSCs proliferation and promotes PSCs differentiation. And the miR-27b target gene, MDFI, promotes PSCs proliferation and inhibits PSCs differentiation in vitro. Furthermore, interfering MDFI expression promotes mice muscle regeneration after injury. CONCLUSION: our results conclude that miR-27b promotes PSCs myogenesis by targeting MDFI. These results expand our understanding of muscle development mechanism in which miRNAs and genes work collaboratively in regulating skeletal muscle development. Furthermore, this finding has implications for obtaining the alternative treatments for patients with the muscle injury.

Mitochondrial Fission Is Required for Angiotensin II-Induced Cardiomyocyte Apoptosis Mediated by a Sirt1-p53 Signaling Pathway.

Hypertension-induced cardiac apoptosis is a major contributor to early-stage heart-failure. Our previous studies have found that p53-mediated mitochondrial fission is involved in aldosterone-induced podocyte apoptosis. However, it is not clear that whether p53-induced mitochondrial fission is critical for hypertensive Angiotensin II (AngII)-induced cardiomyocyte apoptosis. In this study, we found that inhibition of the mitochondrial fission protein Drp1 (dynamin-related protein 1) by Mdivi-1 prevented cardiomyocyte apoptosis and cardiac remodeling in SHRs. In vitro we found that treatment of cultured neonatal rat cardiomyocytes with AngII induced Drp1 expression, mitochondrial fission, and apoptosis. Knockdown of Drp1 inhibited AngII-induced mitochondrial fission and cardiomyocyte apoptosis. Furthermore, AngII induced p53 acetylation. Knockdown of p53 inhibited AngII-induced Drp1 expression, mitochondrial fission, and cardiomyocyte apoptosis. Besides, we found that Sirt1 was able to reverse AngII-induced p53 acetylation and its binding to the Drp1 promoter, which subsequently inhibited mitochondrial fission and eventually attenuated cardiomyocyte apoptosis. Collectively, these results suggest that AngII degrades Sirt1 to increase p53 acetylation, which induces Drp1 expression and eventually results in cardiomyocyte apoptosis. Sirt1/p53/Drp1dependent mitochondrial fission may be a valuable therapeutic target for hypertension induced heart failure.

Erratum: Comparative study of nanostructured carriers of calcium phosphate and magnesium phosphate loaded with SRT1720 for the protection of H2O2-induced senescent endothelium.

[This corrects the article on p. 2068 in vol. 10, PMID: 30093944.].

Therapeutic ultrasound plus pulsed electromagnetic field improves recovery from peripheral arterial disease in hypertension.

The objective of this investigation was to evaluate the therapy effect of combined therapeutic ultrasound (TUS) treatment and pulsed electromagnetic field (PEMF) therapy on angiogenesis in hypertension-related hindlimb ischemia. After subjecting excision of the left femoral artery, spontaneously hypertensive rats (SHRs) were randomly distributed to one of four groups: SHR; TUS treated SHR (SHR-TUS); PEMF treated SHR (PEMF-TUS); and TUS plus PEMF treated SHR (SHR-TUS-PEMF). Wistar-Kyoto rats (WKYs) with femoral artery excision were regarded as a control group. At day 14 after surgery, the TUS plus PEMF united administration had the greatest blood perfusion accompanied by elevated capillary density and the lowest TUNEL index. Interestingly, the united administration up-regulated the angiogenic factors expression of phosphorylated Akt (p-Akt), phosphorylated endothelial nitric oxide synthase (p-eNOS), vascular endothelial growth factor (VEGF), anti-apoptotic protein of Bcl-2 and down-regulated pro-apoptotic protein levels of Bax and Cleaved caspase-3 in vivo. Our results demonstrated that the united administration could significantly rescue hypertension-related inhibition of ischemia-induced neovascularization partly by promoting angiogenesis and inhibiting apoptosis.

Curcumin induces G2/M cell cycle arrest and apoptosis of head and neck squamous cell carcinoma in vitro and in vivo through ATM/Chk2/p53-dependent pathway.

Studies have demonstrated that curcumin (CUR) exerts its tumor suppressor function in a variety of human cancers including head and neck squamous cell carcinoma (HNSCC). However, the exact underlying molecular mechanisms remain obscure. Here, we aim to test whether CUR affects ATM/Chk2/p53 signaling pathway, leading to the induction of cell cycle arrest, inhibition of angiogenesis of HNSCC in vitro and in vivo. To this end, we conducted multiple methods such as MTT assay, Invasion assay, Flow cytometry, Western blotting, RT-PCR, and transfection to explore the functions and molecular insights of CUR in HNSCC. We observed that CUR significantly induced apoptosis and cell cycle arrest, inhibited angiogenesis in HNSCC. Mechanistically, we demonstrated that CUR markedly up-regulated ATM expression and subsequently down-regulated HIF-1α expression. Blockage of ATM production totally reversed CUR induced cell cycle arrest as well as anti-angiogenesis in HNSCC. Moreover, our results demonstrated that CUR exerts its antitumor activity through targeting ATM/Chk2/p53 signal pathway. In addition, the results of xenograft experiments in mice were highly consistent with in vitro studies. Collectively, our findings suggest that targeting ATM/Chk2/p53 signal pathway by CUR could be a promising therapeutic approach for HNSCC prevention and therapy.

Therapeutic ultrasound protects HUVECs from ischemia/hypoxia-induced apoptosis via the PI3K-Akt pathway.

Background: Previous studies have demonstrated that therapeutic ultrasound (TUS) ameliorates angiogenesis on ischemic hind limb animals and also promotes human umbilical vein endothelial cells (HUVECs) tube formation. Apoptosis plays a key role in post-ischemic angiogenesis pathogenesis. However, the mechanisms underlying the anti-apoptotic effects of TUS are not clear. Therefore we put forward the hypothesis that TUS might promote angiogenesis during ischemia/hypoxia (I/H) by decreasing apoptosis. Methods: We investigated the cytoprotective role of TUS and the underlying mechanisms in I/H-induced HUVEC apoptosis. HUVECs were treated under hypoxic serum-starved conditions for 36 h and then treated with or without TUS (9 minutes, 1 MHz, 0.3 W/cm2). The cell viability was examined by the CCK-8 assay, apoptosis cell rate was determined by TUNEL staining and flow cytometry assay. In addition, the mitochondrial-dependent apoptosis pathway was evaluated by the protein activity of Bax, Bcl-2 and Caspase-3. Results: 1) apoptosis could be induced by I/H in HUVECs. 2) TUS attenuates HUVECs cell apoptosis induced by I/H. 3) TUS inhibits the protein expression of apoptosis modulators and effectors that regulate the mitochondrial pathway of apoptosis in HUVECs. 4) TUS increases the phosphorylation of Akt, which demonstrates the activation of the phosphoinositide 3-kinase (PI3K)- serine/threonine kinase (Akt) signal pathway. Conclusions: The present study indicates that exposure to TUS exerts a protective effect against I/H-induced apoptosis among HUVECs and that this process is mediated through the mitochondrial-dependent intrinsic apoptotic pathway. We also confirm that the PI3K-Akt signal cascade may be taken part in the TUS effects on apoptosis.