Tracking mitochondrial Cu(I) fluctuations through a ratiometric fluorescent probe in AD model cells: Towards understanding how AßOs induce mitochondrial Cu(I) dyshomeostasis.

Mitochondrial copper signaling pathway plays a role in Alzheimer's disease (AD), especially in relevant Amyloid-ß oligomers (AßOs) neurotoxicity and mitochondrial dysfunction. Clarifying the relationship between mitochondrial copper homeostasis and both of mitochondrial dysfunction and AßOs neurotoxicity is important for understanding AD pathogenesis. Herein, we designed and synthesized a ratiometric fluorescent probe CHC-NS4 for Cu(I). CHC-NS4 possesses excellent ratiometric response, high selectivity to Cu(I) and specific ability to target mitochondria. Under mitochondrial dysfunction induced by oligomycin, mitochondrial Cu(I) levels gradually increased, which may be related to inhibition of ATP7A-mediated Cu(I) exportation and/or high expression of COX. On this basis, CHC-NS4 was further utilized to visualize the fluctuations of mitochondrial Cu(I) levels during progression of AD model cells induced by AßOs. It was found that mitochondrial Cu(I) levels were gradually elevated during the AD progression, which depended on not only AßOs concentration but also incubation time. Moreover, endocytosis maybe served as a prime pathway mode for mitochondrial Cu(I) dyshomeostasis induced by AßOs during AD progression. These results have provided a novel inspiration into mitochondrial copper biology in AD pathogenesis.

Can Concurrent Fibrate Use Reduce Cardiovascular Risks among Moderate Chronic Kidney Disease Patients Undergoing Statin Therapy? A Cohort Study.

The role of fibrates in treating hypertriglyceridemia in chronic kidney disease (CKD) patients to prevent cardiovascular disease (CVD) has been insufficiently investigated. Since statin is considered the first-line treatment for dyslipidemia in CKD patients, this study aims to evaluate the role of concurrent fibrate therapy with statins among moderate CKD patients. We recruited CKD3 patients from the Chang Gung Research Database who were receiving statin treatment but had not previously been administered ezetimibe or niacin. The participants were divided into two groups based on their use of fibrates (fibrate group) or those with triglyceride levels >200 mg/dL without fibrate treatment (non-fibrate group). The fibrate group (n = 954) only exhibited a significantly lower incidence of AMI (4.4% vs. 5.4%, HR: 0.77, 95% CI: 0.61 to 0.98). The risk of major adverse cardiovascular and cerebrovascular events (14.7% vs. 15.6%, HR: 0.91, 95% CI: 0.72 to 1.15) and all-cause mortality (5.7% vs. 6.1%, HR: 0.91, 95% CI: 0.63 to 1.30) did not significantly differ between the fibrate group and the non-fibrate group (n = 2358). In moderate CKD patients, combining fibrate therapy with statins may not offer additional cardiovascular protection compared to statin alone.

Effects of Suramin on Polycystic Kidney Disease in a Mouse Model of Polycystin-1 Deficiency.

The aberrant activation of the purinergic signaling pathway has been shown to promote cyst growth and fluid secretion in autosomal dominant polycystic kidney disease (ADPKD). Suramin is an anti-parasitic drug that has strong anti-purinergic properties. Whether suramin could have a therapeutic effect on ADPKD has not been fully investigated. We examined the effect of suramin on cyst progression in a Pkd1 microRNAs transgenic mouse model that presented stable Pkd1 knockdown and moderate disease progression. The Pkd1-deficient mice were treated with suramin (60 mg/kg) by intraperitoneal injection twice a week from postnatal days 35 to 90. Kidney-to-body weight ratios, cyst indices, and blood urea nitrogen (BUN) levels were measured. Cell proliferation and macrophage infiltration were determined by immunohistochemistry. The suramin-treated group had significantly lower renal cyst densities, cell proliferation, and macrophage infiltration compared with saline-treated controls. Suramin significantly inhibited ERK phosphorylation and the expression of Il1b, Il6, Nlrp3, Tgfb, Fn1, P2rx7, and P2ry2 mRNAs in the kidneys. However, BUN levels remained high despite the reduction in cyst growth. Furthermore, plasma cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) levels were significantly higher in the suramin-treated group compared with the control group. Periodic acid-Schiff staining revealed degenerative changes and epithelial cell vacuolation in the non-cystic renal tubules, which indicated phospholipidosis following suramin treatment. These results suggest that suramin may reduce renal cyst growth and inflammation, but the associated tubular cell injuries could limit its therapeutic potential. Other purinergic receptor antagonists with less nephrotoxicity may deserve further investigation for the treatment of ADPKD.

Customized Photothermal Therapy of Subcutaneous Orthotopic Cancer by Multichannel Luminescent Nanocomposites.

Photothermal therapy (PTT) is a potentially advanced strategy for highly precise cancer treatment. Tumor-microenvironment-activatable agents provide useful tools for PTT, but their photothermal conversion capacities vary and cannot be evaluated in vivo; thus, a general PTT prescription does not work with individual activatable agents. Here, glutathione (GSH)-activatable nanocomposites, silicomolybdate-functionalized NaLuF4 :Yb,Er@NaLuF4 @NaLuF4 :Nd are prepared, for customized PTT of subcutaneous orthotopic cancer. By simultaneously determining intratumoral GSH concentration and the amount of accumulated agent using multiple orthogonal luminescent emissions of nanocomposites, near-infrared absorbance of photothermal conversion agents is evaluated in vivo, based on the optimized irradiating prescriptions (irradiating power density and time) established. This allows customized PTT of each individual case with high efficacy and viability. This work also includes a method for investigating individual intratumoral variation, and the development of the next generation of customized nanomedicine for efficacious PTT of subcutaneous orthotopic cancer.

Orthogonal Near-Infrared-II Imaging Enables Spatially Distinguishing Tissues Based on Lanthanide-Doped Nanoprobes.

Multichannel near-infrared (NIR)-II imaging provides more precise and detailed information for studying complex biological processes. When studying specific biological processes, a separated single signal and multisignals are essential but difficult to obtain by traditional multichannel NIR-II imaging methods. Taking advantage of the unique optical properties of lanthanide ions, especially in atom-like absorbance and emission spectroscopy in the NIR region, in this study, we synthesized two lanthanide-doped nanoprobes, NaYF4:Gd@NaYF4:Nd@NaYF4 (cssNd) and NaYF4:Gd@NaYF4:Er@NaYF4 (cssEr). These two nanoprobes show orthogonal NIR-II emissions (1064 and 1330 nm for cssNd and 1550 nm for cssEr) under 730 and 980 nm excitation, respectively. The feasibility of cssNd and cssEr for multichannel NIR-II imaging was proven in vitro. Under different methods of administering the nanoprobes, in vivo multichannel NIR-II imaging with both the separated single signal and multisignals was successfully performed and could spatially distinguish tissues under two different excitation sources. Our results provide a new method for multichannel NIR-II imaging with separable signals, which is promising for precisely studying complex biological processes precisely.

A reproduceable in situ xenograft model of spinal glioma.

BACKGROUND: Spinal glioma is a nervous system tumor that tends to relapse and has no specific prognostic molecular biomarkers. Thus, a stable and reproduceable animal research model of spinal glioma is urgently needed. NEW METHOD: We established a new in situ tumor xenograft model of spinal glioma using nude mice. In this study, we implanted tumors into the cervical spinal cord of nude mice to mimic the pathological characteristics of the original tumors. RESULTS: Through anatomical experiments, we found that the cervical lamina of mice was thinner, the intervertebral space was much wider, and the adhesion muscles were more easily separated. According to the examination of spinal cord sections, the best puncture point we identified was located 0.9 mm lateral to the posterior median line at the level of the line between the midpoints of the scapulae and at a depth of 0.9 mm. In the nude mouse xenograft experiment, the implanted tumor tissue retained the pathological characteristics of the original tumor. COMPARISON WITH EXISTING METHOD(S): This model used the cervical spinal cord as the puncture site and patient-derived primary tumor cells, which has never been performed before. Tumor cells could be injected directly without damaging the lamina. Thus, we could reduce the risk of man-made spinal cord injury and infection and avoid destroying the stability and integrity of the spine. CONCLUSIONS: This study established a stable and reliable animal model of spinal glioma for further molecular research and targeted therapy development.

A Novel Classification and Its Clinical Significance in Spinal Schwannoma Based on the Membranous Hierarchy.

BACKGROUND: Spinal schwannoma is a common benign tumor. However, the high recurrence rate and incidence of surgical complications are unsolved problems. OBJECTIVE: To propose a morphological classification of spinal schwannoma based on tumor-membrane relationships to increase the gross total resection (GTR) rate and to decrease the incidence of surgical complications. METHODS: Histological techniques were used to study 7 adult cadavers. Following picrosirius staining, the membranes around the nerve root were observed under a microscope. Data from 101 patients with spinal schwannoma were also collected for clinical analysis. RESULTS: The sleeve around the spinal nerve root consisted of dura and arachnoid tissues. The space between them gradually narrowed and fused at the proximal pole of the nerve root ganglion. Spinal schwannomas were divided into 4 types based on membranous structure: intrapial (type I), subarachnoidal (type II), intra- and extradural (type III), and extradural growth (type IV). Types II and III were further subdivided into 2 subtypes. GTR was achieved in all patients (100%), with no tumor recurrence during follow-up. Overall functional status significantly improved postoperatively. A total of 59 patients (92%) showed improvement or significant improvement postoperatively. There was no difference in surgical outcomes among the tumor classifications (P = .618). No intraoperative vertebral artery injuries or postoperative cerebrospinal fluid fistula occurred. CONCLUSION: Spinal schwannoma classification based on a membranous hierarchy provides an intuitive platform for preoperative planning and intraoperative safety. This classification scheme may help surgeons better define surgical goals and anticipate or even avoid complications from resection.

HMGB1-Induced p62 Overexpression Promotes Snail-Mediated Epithelial-Mesenchymal Transition in Glioblastoma Cells via the Degradation of GSK-3ß.

Rationale: Glioblastoma (GBM) is the most common and aggressive brain tumor, characterized by its propensity to invade the surrounding brain parenchyma. The effect of extracellular high-mobility group box 1 (HMGB1) protein on glioblastoma (GBM) progression is still controversial. p62 is overexpressed in glioma cells, and has been associated with the malignant features and poor prognosis of GBM patients. Hence, this study aimed to clarify the role of p62 in HMGB1-induced epithelial-mesenchymal transition (EMT) of GBM both in vitro and in vivo. Methods: Immunoblotting, immunofluorescence and qRT-PCR were performed to evaluate EMT progression in both human GBM cell line and primary GBM cells. Transwell and wound healing assays were used to assess the invasion and migration of GBM cells. shRNA technique was used to investigate the role of p62 in HMGB1-induced EMT both in vitro and in vivo orthotopic tumor model. Co-immunoprecipitation assay was used to reveal the interaction between p62 and GSK-3ß (glycogen synthase kinase 3 beta). Immunohistochemistry was performed to detect the expression levels of proteins in human GBM tissues. Results: In this study, GBM cells treated with recombinant human HMGB1 (rhHMGB1) underwent spontaneous EMT through GSK-3ß/Snail signaling pathway. In addition, our study revealed that rhHMGB1-induced EMT of GBM cells was accompanied by p62 overexpression, which was mediated by the activation of TLR4-p38-Nrf2 signaling pathway. Moreover, the results demonstrated that p62 knockdown impaired rhHMGB1-induced EMT both in vitro and in vivo. Subsequent mechanistic investigations showed that p62 served as a shuttling factor for the interaction of GSK-3ß with proteasome, and ultimately activated GSK-3ß/Snail signaling pathway by augmenting the degradation of GSK-3ß. Furthermore, immunohistochemistry analysis revealed a significant inverse correlation between p62 and GSK-3ß, and a combination of the both might serve as a more powerful predictor of poor survival in GBM patients. Conclusions: This study suggests that p62 is an effector for HMGB1-induced EMT, and may represent a novel therapeutic target in GBM.

HERC3-Mediated SMAD7 Ubiquitination Degradation Promotes Autophagy-Induced EMT and Chemoresistance in Glioblastoma.

PURPOSE: Glioblastoma, a common malignant intracranial tumor, has the most dismal prognosis. Autophagy was reported to act as a survival-promoting mechanism in gliomas by inducing epithelial-to-mesenchymal transition (EMT). Here, we determined the critical molecules involved in autophagy-induced EMT and elucidated the possible mechanism of chemoradiotherapy resistance and tumor recurrence. EXPERIMENTAL DESIGN: We used isobaric tags for relative and absolute quantitation to identify the critical proteins and pathway mediating EMT via autophagy inducer treatment, and tested the expression of these proteins using tissue microarray of gliomas and clinical glioblastoma samples as well as tissues and cells separated from the core lesion and tumor-peripheral region. Analysis of the Cancer Genome Atlas database and 110 glioblastoma cases revealed the prognostic value of these molecules. The functional role of these critical molecules was further confirmed by in vitro experiments and intracranial xenograft in nude mice. RESULTS: Autophagy inducers significantly upregulated the expression of HERC3, which promotes ubiquitination-mediated degradation of SMAD7 in an autolysosome-dependent manner. The corresponding increase in p-SMAD2/3 level and TGFß pathway activation finally induced EMT in cell lines and primary glioblastoma cells. Moreover, HERC3 overexpression was observed in pseudo-palisade cells surrounding tumor necrosis and in tumor-adjacent tissue; high HERC3 and low SMAD7 levels predicted poor clinical outcome in glioblastoma; xenograft of nude mice and in vitro experiments confirmed these findings. CONCLUSIONS: Together, our findings reveal the indispensable role of HERC3 in regulating canonical SMAD2/3-dependent TGFß pathway involvement in autophagy-induced EMT, providing insights toward a better understanding of the mechanism of resistance to temozolomide and peripheral recurrence of glioblastoma.

Biocompatible Heat-Shock Protein Inhibitor-Delivered Flowerlike Short-Wave Infrared Nanoprobe for Mild Temperature-Driven Highly Efficient Tumor Ablation.

Multifunctional nanomaterials for dual-mode imaging guided cancer therapy are highly desirable in clinical applications. Herein, a flowerlike NiS2-coated NaLuF4:Nd (Lu:Nd@NiS2) nanoparticle was synthesized as a novel therapeutic agent for short-wave infrared light imaging and magnetic resonance imaging to guide photothermal therapy (PTT). The material was then loaded with phenolic epigallocatechin 3-gallate (EGCG), which is a natural heat-shock protein 90 (HSP90) inhibitor. Upon near infrared irradiation, EGCG was released from the Lu:Nd@NiS2-EGCG, which bound HSP90 and reduced cell tolerance to heat, resulting in a better therapeutic effect at the same elevated temperature. Therefore, with minimal side effects and remarkable antitumor efficacy in vivo, Lu:Nd@NiS2-EGCG appeared to be a promising photothermal agent for enhanced PTT.

Green synthesis of ultra-small VOx nanodots for acidic-activated HSP60 inhibition and therapeutic enhancement.

Using metal oxide semiconductor nanomaterials for synergistic cancer treatment has recently attracted the attention of numerous researchers. Herein, oxygen-defective vanadium oxide nanodots (VOx NDs) with ultra-small size and great dispersibility were synthesized via a novel user-friendly method, and then doxorubicin was loaded onto the VOx NDs surfaces. The VOx NDs had great photothermal conversion efficiency and stability. Doxorubicin-loaded VOx NDs can simultaneously serve as therapeutic agent and tumor microenvironment-activable HSP60 inhibitor, resulting in improved efficacy of photothermal therapy and released active doxorubicin for chemotherapy. Finally, we show that synergistic treatment achieved significant therapeutic effects in mice. These results provided a promising strategy for developing novel methods of synthesizing metal oxide semiconductors for enhanced synergistic cancer treatment.

Rationally designed upconversion nanoprobe for simultaneous highly sensitive ratiometric detection of fluoride ions and fluorosis theranostics.

For many years, fluorosis has been known as a worldwide disease which seriously diminishes quality of life through skeletal embrittlement and hepatic damage. Aiming to develop novel drugs for simultaneous fluorosis diagnosis and therapy, in this work we explore the feasibility of a novel pyrogallic acid-titanium(iv) complex-modified upconversion nanoprobe (UCNP-PA-Ti) for F- capture and real-time quantification. Utilizing the strong interaction between Ti4+ and F-, the modified PA-Ti decomposes in F--containing solution, which not only weakens the FRET but results in upconversion luminescence (UCL) recovery. Both in vitro and in vivo experiments demonstrate a highly sensitive F- UCL response and therapeutic efficiency, which was promising for successful UCL image monitoring and the therapeutic process. Long blood circulation time and low toxicity ensured their safe application for fluorosis theranostics. Our work provides a new possibility for F- concentration detection within fluorosis therapeutic periods and encourages the development of novel drugs for fluorosis theranostics.

Policosanol fabrication from insect wax and optimization by response surface methodology.

BACKGROUND: Insect wax is a famous biological resource for the role in economic production in China. Insect wax is a good source of policosanol, which may is a candidate supplement in foodstuff and pharmaceuticals that has important physiological activities. Therefore, this work aims to investigate a high-yield and rapid method for policosanol fabrication from insect wax. RESULTS: The conditions for policosanol fabrication were optimized as follows: an oil bath temperature of 112.7°C and reductant dosage of 0.97 g (used for the reduction of 10.00 g of insect wax). The yield of policosanol reached 83.20%, which was 4 times greater than that of existing methods, such as saponification. The total content of policosanol obtained under the optimal conditions reached 87%. In other words, a high yield of policosanol was obtained from insect wax (723.84 mg/g), that was 55 times higher than that generated from beeswax-brown via saponification. The concentrations of metal residues in policosanol were within the limits of the European Union regulations and EFSA stipulation. The LD50 values for oral doses of insect wax and policosanol were both > 5 g/kg. CONCLUSION: Policosanol was fabricated via solvent-free reduction from insect wax using LiAlH4 at a high yield. The fabrication conditions were optimized. Policosanol and insect wax showed high security, which made them potential candidates as supplements in foods, pharmaceuticals and cosmetics. The rapid and high-yield method has great potential for commercial manufacturing of policosanol.

In vivo evaluation of insect wax for hair growth potential.

Insect wax is secreted by Ericerus pela Chavanness. It has been traditionally used to treat hair loss in China, but few reports have been published on the hair growth-promoting effect of insect wax. In this work, we examined the hair growth-promoting effects of insect wax on model animals. Different concentrations of insect wax were topically applied to the denuded backs of mice, and 5% minoxidil was applied topically as a positive control. We found that insect wax significantly promoted hair growth in a dose-dependent manner, 45% and 30% insect wax both induced hair to regrow, while less visible hair growth was observed in blank controls on the 16th day. The experimental areas treated with 45% and 30% insect wax exhibited significant differences in hair scores compared to blank controls, and hair lengths in the 45% and 30% insect wax group was significantly longer than in blank controls on the 16th and 20th days. There were no new hair follicles forming in the treated areas, and the hair follicles were prematurely converted to the anagen phase from the telogen phase in experimental areas treated with 45% and 30% insect wax. Both 45% and 30% insect wax upregulated vascular endothelial growth factor expression. The results indicated that 45% and 30% insect wax showed hair growth-promoting potential approximately as potent as 5% minoxidil by inducing the premature conversion of telogen-to-anagen and by prolonging the mature anagen phase rather than increasing the number of hair follicles, which was likely related to the upregulation of VEGF expression. The dissociative policosanol in insect wax was considered the key ingredient most likely responsible for the hair growth promoting potential.

Artemisinin-Loaded Mesoporous Nanoplatform for pH-Responsive Radical Generation Synergistic Tumor Theranostics.

The development of novel and effective cancer treatments will greatly contribute to prolonging and improving patient lives. In this study, a multifunctional nanoplatform was designed and developed based on mesoporous NiO (mNiO) nanoparticles and terbium complexes as an artemisinin (ART) vehicle, a T2-weighted contrast agent, and a luminescence imaging probe. mNiO is a novel pH-responsive material that can degrade and release nickel ions (Ni2+) in an acidic tumor microenvironment. The endoperoxide bridge bond in the structure of ART tends to react with Ni2+ to produce radicals that can kill tumor cells. On the basis of its excellent near-infrared absorbance, mNiO can also be considered as a novel photothermal conversion agent for cancer photothermal therapy (PTT). Compared with free ART or PTT only, this novel agent showed remarkably enhanced antitumor activity in cultured cells and in tumor mice models, owing to the hypoxic tumor microenvironment impelling synergistic therapeutic action. These results provide a novel way of using a promising natural drug-based nanoplatform for synergistic therapy of tumors.

Hair growth promoting effect of white wax and policosanol from white wax on the mouse model of testosterone-induced hair loss.

White wax (WW) has been traditionally used to treat hair loss in China. However there has been no reporter WW and its extract responsible for hair growth-promoting effect on androgenetic alopecia. In this paper, we examined the hair growth-promoting effects of WW and policosanol of white wax (WWP) on model animal of androgenetic alopecia and the potential target cell of WW and WWP. WW (1, 10 and 20%) and WWP (0.5, 1 and 2%) were applied topically to the backs of mice. Finasteride (2%) was applied topically as a positive control. MTS assays were performed to evaluate cell proliferation in culture human follicle dermal papilla cells (HFDPCs). The inhibition of WW and WWP for 5α- reductase were tested in Vitro. Results showed more lost hairs were clearly seen in mice treated with TP only and TP plus vehicle. Mice which received TP plus WW and WWP showed less hair loss. WW and WWP showed an outstanding hair growth-promoting activity as reflected by the follicular length, follicular density, A/T ratio, and hair bulb diameter. The optimal treatment effect was observed at 10% WW and 1% WWP, which were better than 2% finasteride treatment. MTS assay results suggested that WW and WWP remarkably increased the proliferation of HFDPCs. Inhibitor assay of 5α- reductase showed that WW and WWP inhibited significantly the conversion of testosterone to dihydrotesterone, and the IC50 values of WW and WWP were higher than that of finasteride. In Conclusion, WW and WWP could act against testosterone-induced alopecia in mice, and they promoted hair growth by inhibiting 5α-reductase activity and HFDPCs proliferation. DPCs is the target cell of WW and WWP.

Artificially controlled degradable inorganic nanomaterial for cancer theranostics.

Multifunctional nanomaterials for cancer diagnosis and therapy have recently prompted widespread concern. To avoid nanotoxicity, the development of novel degradable functional materials must be our main focus. In this study, we firstly developed ethylenediaminetetraacetic acid calcium disodium salt (EDTA)- and bovine serum albumin (BSA)-capped Mn3O4 nanoparticles (MONPs-BSA-EDTA) as a novel inorganic nanomaterials for multifunctional imaging-guided photothermal therapy, which can be degraded in a progress-controlled way by artificially introduced ascorbic acid. The degradation products can also be captured and their excretion accelerated. Careful studies suggested that the toxicity of the MONPs-BSA-EDTA and its degradation products is low. The degradation mechanism also suggests a new method of controlled drug release. The development of artificially controlled degradable inorganic nanomaterials also provides a new way to degrade nanomaterials and minimize ion release, which may have potential applications in cancer theranostics without nanotoxicity.

Polydopamine-Encapsulated Fe3O4 with an Adsorbed HSP70 Inhibitor for Improved Photothermal Inactivation of Bacteria.

Photothermal treatment, a new approach for inactivation of bacteria and pathogens that does not depend on traditional therapeutic approaches, has recently received much attention. In this study, a new type of nanoplatform (PDA@Fe3O4 + PES) was fabricated by using polydopamine (PDA, a photothermal conversion agent) to encapsulate Fe3O4 (a magnetic nanoparticle) and support 2-phenylethynesulfonamide (PES, an inhibitor of heat shock protein 70 (HSP70)). Upon near-infrared light irradiation, the increased temperature weakens π-π and hydrogen bonding interactions, and PES is released from the PDA@Fe3O4 + PES. The released PES inhibits the function of HSP70, reducing bacterial tolerance to photothermal therapy and improving the therapeutic effect against infectious bacterial pathogens. After treatment, PDA@Fe3O4 + PES can be recovered using the magnetic property of the Fe3O4 cores. Consequently, PDA@Fe3O4 + PES possesses the potential to be a recyclable photothermal agent for enhanced photothermal bacterial inactivation without causing secondary pollution.