Moscatilin Inhibits Metastatic Behavior of Human Hepatocellular Carcinoma Cells: A Crucial Role of uPA Suppression via Akt/NF-κB-Dependent Pathway.

Hepatocellular carcinoma (HCC) frequently shows early invasion into blood vessels as well as intrahepatic metastasis. Innovations of novel small-molecule agents to block HCC invasion and subsequent metastasis are urgently needed. Moscatilin is a bibenzyl derivative extracted from the stems of a traditional Chinese medicine, orchid Dendrobium loddigesii. Although moscatilin has been reported to suppress tumor angiogenesis and growth, the anti-metastatic property of moscatilin has not been elucidated. The present results revealed that moscatilin inhibited metastatic behavior of HCC cells without cytotoxic fashion in highly invasive human HCC cell lines. Furthermore, moscatilin significantly suppressed the activity of urokinase plasminogen activator (uPA), but not matrix metalloproteinase (MMP)-2 and MMP-9. Interestingly, moscatilin-suppressed uPA activity was through down-regulation the protein level of uPA, and did not impair the uPA receptor and uPA inhibitory molecule (PAI-1) expressions. Meanwhile, the mRNA expression of uPA was inhibited via moscatilin in a concentration-dependent manner. In addition, the expression of phosphorylated Akt, rather than ERK1/2, was inhibited by moscatilin treatment. The expression of phosphor-IκBα, and -p65, as well as κB-luciferase activity were also repressed after moscatilin treatment. Transfection of constitutively active Akt (Myr-Akt) obviously restored the moscatilin-inhibited the activation of NF-κB and uPA, and cancer invasion in HCC cells. Taken together, these results suggest that moscatilin impedes HCC invasion and uPA expression through the Akt/NF-κB signaling pathway. Moscatilin might serve as a potential anti-metastatic agent against the disease progression of human HCC.

Evaluating Molecular Properties Involved in Transport of Small Molecules in Stratum Corneum: A Quantitative Structure-Activity Relationship for Skin Permeability.

The skin permeability (Kp) defines the rate of a chemical penetrating across the stratum corneum. This value is widely used to quantitatively describe the transport of molecules in the outermost layer of epidermal skin and indicate the significance of skin absorption. This study defined a Kp quantitative structure-activity relationship (QSAR) based on 106 chemical substances of Kp measured using human skin and interpreted the molecular interactions underlying transport behavior of small molecules in the stratum corneum. The Kp QSAR developed in this study identified four molecular descriptors that described the molecular cyclicity in the molecule reflecting local geometrical environments, topological distances between pairs of oxygen and chlorine atoms, lipophilicity, and similarity to antineoplastics in molecular properties. This Kp QSAR considered the octanol-water partition coefficient to be a direct influence on transdermal movement of molecules. Moreover, the Kp QSAR identified a sub-domain of molecular properties initially defined to describe the antineoplastic resemblance of a compound as a significant factor in affecting transdermal permeation of solutes. This finding suggests that the influence of molecular size on the chemical's skin-permeating capability should be interpreted with other relevant physicochemical properties rather than being represented by molecular weight alone.

CAN008 prolongs overall survival in patients with newly diagnosed GBM characterized by high tumor mutational burden.

BACKGROUND: A previous phase 1 dose-escalation study in Taiwan indicated CAN008 (asunercept) with standard concurrent chemoradiotherapy (CCRT) improved progression-free survival (PFS) in newly diagnosed glioblastoma (GBM) patients. This study evaluates the efficacy of CAN008 in promoting overall survival (OS) and identifies genetic alterations associated with treatment responses. METHODS: We compared OS of 5-year follow-ups from 9 evaluable CAN008 cohort patients (6 received high-dose and 3 received low-dose) to a historical Taiwanese GBM cohort with 164 newly diagnosed patients. CAN008 treatment response-associated genetic alterations were identified by whole-exome sequencing and comparing variant differences between response groups. Associations among patient survival, tumor mutational burden (TMB), and genetic alterations were analyzed using CAN008 cohort and TCGA-GBM dataset. RESULTS: OS for high-dose CAN008 patients at 2 and 5 years was 83% and 67%, respectively, and 40.1% and 8.8% for the historical GBM cohort, respectively. Better OS was observed in the high-dose CAN008 cohort (without reaching the median survival) than the historical GBM cohort (median OS: 20 months; p=0.0103). Five high-dose CAN008 patients were divided into good and poor response groups based on their PFS. A higher variant count and TMB were observed in good response patients, whereas no significant association was observed between TMB and patient survival in the newly diagnosed TCGA-GBM dataset, suggesting TMB may modulate patient CAN008 response. CONCLUSION: CAN008 combined with standard CCRT treatment prolonged the PFS and OS of newly diagnosed GBM patients compared to standard therapy alone. Higher treatment efficacy was associated with higher TMB.

Solution-phase parallel synthesis and screening of anti-tumor activities from fenbufen and ethacrynic acid libraries.

The derivatives with fenbufen and ethacrynic acid core compounds was synthesized through a facial preparation of 1-amino-4-azidobutane. The subsequent coupling with 102 members of carboxylic acids afforded amide products. The in situ screening using colorimetric assay with 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide showed that fenbufen but not ethacrynic acid butyl amide members displayed the cytotoxicities to tumor cells substantially, including two human cell lines (MCF7 and A549) and two murine cell lines (C26 and TRAMP-C1). Three fenbufen analogs were found to have a good anti-tumor activity comparable to cisplatin.

Synthesis and structure-activity relationships of fenbufen amide analogs.

The previous discoveries of butyl fenbufen amide analogs with antitumor effects were further examined. The amide analogs with 1, 3, 4 and 8 carbons chains were prepared in 70-80% yield. Fenbufen had no cytotoxic effects at concentrations ranging from 10 to 100 µM. Methyl fenbufen amide had significant cytotoxic effects at a concentration of 100 µM. As the length of the alkyl amide side chain increased, the cytotoxic effects increased, and the octyl fenbufen amide had the greatest cytotoxic effect. After treatment with 30 µM octyl fenbufen amide, nearly seventy percent of the cells lost their viability. At the concentration of 10 µM, fenbufen amide analogs did not show cytotoxicity according to the MTT assay results. The NO scavenging activities of the fenbufen amide analogs were not significantly different from those of fenbufen.

Inflammatory pseudotumor of the sinonasal tract.

Inflammatory pseudotumors (IPTs) are a clinically and histologically diverse group of lesions characterized by a tumor mass of acute and chronic inflammatory cells with a variable fibrous response. IPTs most commonly involve the lung and orbit, but rarely the sinonasal tract. We report a 68-year-old male with an IPT of the sinonasal tract presenting as nasal obstruction and postnasal dripping for several years. A gray-white soft mass was noted in the right nasal cavity. Computed tomography revealed a solid mass filling the right nasal cavity and maxillary sinus. Caldwell-Luc operation with ethmoidectomy (right) was conducted to resect the mass en bloc. Pathology revealed admixture of plasma cells, lymphocytes and eosinophils, confirming the diagnosis of IPT. The patient remained symptom-free over 4 years of follow-up. Awareness of the clinical presentations, histopathologic features and treatment of choice of this rare disease entity is necessary to distinguish it from malignancy and avoid unnecessary management.

Release of Doxorubicin by a Folate-Grafted, Chitosan-Coated Magnetic Nanoparticle.

In clinical tumor therapy, chemotherapeutic routes have caused severe side effects; current delivery methods are unsatisfactory. Successful design of a remotely folate (FA)-grafted chitosan (CS)-coated magnetic nanoparticle (MNP) with low toxicity, has been achieved. A chemotherapeutic drug such as doxorubicin (DOX), is loaded in the MNP-based matrix (FA-grafted CS-DOX-TPP-MNP), which is coated by an activated target tumor molecule of FA-grafted CS biopolymer with the inclusion of tripolyphosphate (TPP) as a linker. The resultant nano-complexes exhibited random aggregates (~240 nm) and zeta potential (-24.9 mV). In vivo experiments using athymic BALB/c nude mice with human glioblastoma U87 cells in a subcutaneous tumor model revealed that magnetic guidance of FA-grafted CS-DOX-TPP-MNP, injected via the tail vein, significantly decreased tumor growth. This manuscript demonstrates the feasibility of magnetizing control of FA-grafted CS-DOX-TPP-MNP to enhance the localization of drug release.

Quantitative Measurement of the Thyroid Uptake Function of Mouse by Cerenkov Luminescence Imaging.

Cerenkov luminescence imaging (CLI) has been an evolutional and alternative approach of nuclear imaging in basic research. This study aimed to measure the 131I thyroid uptake of mouse using CLI for assessment of thyroid function. Quantification of 131I thyroid uptake of mice in euthyroid, hypothyroid and hyperthyroid status was performed by CLI and γ-scintigraphy at 24 hours after injection of 131I. The 131I thyroid uptake was calculated using the equation: (thyroid counts - background counts)/(counts of injected dose of 131I) × 100%. Serum T4 concentration was determined to evaluate the thyroid function. The radioactivity of 131I was linearly correlated with the CL signals in both in vitro and in vivo measurements. CLI showed a significant decrease and increase of 131I thyroid uptake in the mice in hypo- and hyperfunctioning status, respectively, and highly correlated with that measured by γ-scintigraphy. However, the percent thyroid uptake measured by CLI were one-fifth of those measured by γ-scintigraphy due to insufficient tissue penetration of CL. These results indicate that CLI, in addition to nuclear imaging, is able to image and evaluate the 131I thyroid uptake function in mice in preclinical and research settings.

Submandibular cavernous hemangiomas with multiple phleboliths masquerading as sialolithiasis.

Vasoformative tumors (almost exclusively hemangiomas) are the most common lesions of the major salivary glands during infancy and early childhood. They are more common in the parotid gland but are particularly rare in the submandibular gland. Changes in blood flow dynamics within hemangiomas result in thrombus formation and phleboliths. Hemangiomas of the salivary glands in adults are histologically unlike those in infants, the former being characteristically of the cavernous variant. Most cavernous hemangiomas require surgery since they do not show a tendency to regress. A case of an adult man with cavernous hemangioma affecting the submandibular salivary gland that clinically simulated sialolithiasis is presented to alert surgeons to the possibility of such a lesion. We describe the clinical course and review the literature.

Diagnosing minimal adenocarcinoma on prostate needle biopsy by real-time dynamic telepathology through the internet: evaluation of an economic technology for remote consultation.

Computer-aided telepathology was introduced about 10 years ago, but has not yet met with worldwide acceptance. Recently, the internet has been used for image transmission in telepathology. We set up an easily assembled system comprising a common microscope, a charge-coupled device (CCD) camera, a personal computer, and a commercial internet surveillance program with internet accessibility. The consultant then views the real-time images using a common web browser at the remote site. The purpose of the study was to assess the ability of the system to transmit images of sufficient quality to achieve high concordance between the diagnoses made at the home base and at the remote site. We chose cases of minimal adenocarcinoma on prostate needle biopsy, because these lesions are liable to be overlooked and, even if discovered, are subject to differences in interpretation due to their limited size and subtle histologic changes. One hundred prostate needle biopsy specimens, including 45 minimal adenocarcinoma, 11 atypical small acinar proliferation, and 44 benign lesions, were tested. Two pathologists, unaware of the final diagnoses, were recruited to provide intra- and interinstitutional consultation. The overall concordance rates between telepathology diagnoses and final diagnoses were 97% and 94% for the two pathologists, respectively. Our results demonstrate that this method is effective for teleconsultation. Similar systems using the internet can be easily set up by ordinary pathology laboratories to facilitate remote consultation.

Renal tubular dysgenesis in siblings.

Renal tubular dysgenesis is a recently recognized autosomal recessive condition characterized by absence or poor development of proximal convoluted tubules, clinical presentation of oligohydromnios, Potter sequence, and neonatal respiratory failure. Only a few cases of renal tubular dysgenesis have been diagnosed. We report 2 cases from a single family. Histologically, the renal cortices showed crowding of the glomeruli and primitive tubules, most of which demonstrated positive cytoplasmic staining for epithelial membrane antigen and peanut lectin. Electron microscopy of the kidneys of the second baby revealed absence of differentiated characteristics of the proximal tubules.

Gadolinium-functionalized nanographene oxide for combined drug and microRNA delivery and magnetic resonance imaging.

The delivery of anti-cancer therapeutics to tumors at clinically effective concentrations, while avoiding nonspecific toxicity, remains a major challenge for cancer treatment. Here we present nanoparticles of poly(amidoamine) dendrimer-grafted gadolinium-functionalized nanographene oxide (Gd-NGO) as effective carriers to deliver both chemotherapeutic drugs and highly specific gene-targeting agents such as microRNAs (miRNAs) to cancer cells. The positively charged surface of Gd-NGO was capable of simultaneous adsorption of the anti-cancer drug epirubicin (EPI) and interaction with negatively charged Let-7g miRNA. Using human glioblastoma (U87) cells as a model, we found that this conjugate of Let-7g and EPI (Gd-NGO/Let-7g/EPI) not only exhibited considerably higher transfection efficiency, but also induced better inhibition of cancer cell growth than Gd-NGO/Let-7g or Gd-NGO/EPI. The concentration of Gd-NGO/Let-7g/EPI required for 50% inhibition of cellular growth (IC50) was significantly reduced (to the equivalent of 1.3 µg/mL EPI) compared to Gd-NGO/EPI (3.4 µg/mL EPI). In addition, Gd-NGO/Let-7g/EPI could be used as a contrast agent for magnetic resonance imaging to identify the location and extent of blood-brain barrier opening and quantitate drug delivery to tumor tissues. These results suggest that Gd-NGO/Let-7g/EPI may be a promising non-viral vector for chemogene therapy and molecular imaging diagnosis in future clinical applications.

Compartment syndrome following robotic-assisted prostatectomy: rhabdomyolysis in bone scintigraphy.

Major urologic surgery performed in the lithotomy position sometimes results in the serious complication of rhabdomyolysis. A 56-year-old man was admitted to the hospital for prostate adenocarcinoma. A whole-body bone scan was performed to exclude bony metastases, which demonstrated no bone lesions but showed intense soft-tissue activity in gluteus maximus muscles, findings suggestive of a myopathy. He had just undergone right nerve-sparing radical prostatectomy in the lithotomy position for 6 hours and presented with swollen bilateral thighs. Elevation of creatine kinase level confirmed muscle injury.

The use of cationic microbubbles to improve ultrasound-targeted gene delivery to the ischemic myocardium.

We synthesized a cationic microbubble (CMB) with the aim of enhancing its DNA-carrying capacity to improve targeted gene transfection of the ischemic heart for cardiac regeneration. We previously reported that ultrasound-targeted microbubble destruction (UTMD) employing the commercial Definity microbubble (MB) successfully transfected genes into rodent hearts, but the transfection efficiency was modest. We synthesized a CMB and compared its DNA-carrying capacity and reporter gene transfection efficiency with the Definity MB. The CMB bound 70% more plasmid DNA than the Definity MB. UTMD-mediated gene delivery with the CMB enhanced both transfection efficiency and gene expression. In vivo studies assessed the ability of the CMB to deliver the therapeutic AKT gene to the ischemic rat myocardium and evaluated the effects on apoptosis, angiogenesis, and cardiac function. AKT transfection with the CMB reduced infarct size (p < 0.05), increased infarct thickness (p < 0.05), reduced apoptosis (p < 0.05), increased vascular density (p < 0.05), and improved cardiac perfusion and function (p < 0.05) compared to the Definity MB. Delivery of AKT with the CMB resulted in greater cardiac functional improvements compared to the Definity MB. UTMD therapy with this CMB provides an efficient platform for the targeted delivery of factors required to regenerate the ischemic heart and preserve cardiac function.

Study of [18F]FLT and [123I]IaraU for cellular imaging in HSV1 tk-transfected murine fibrosarcoma cells: evaluation of the tracer uptake using 5-fluoro, 5-iodo and 5-iodovinyl arabinosyl uridines as competitive probes.

As one of the most intensively studied probes for imaging of the cellular proliferation, [(18)F]FLT was investigated whether the targeting specificity of thymidine kinase 1 (TK1) dependency could be enhanced through a synergistic effect mediated by herpes simplex type 1 virus (HSV1) tk gene in terms of the TK1 or TK2 expression. 5-[(123)I]Iodo arabinosyl uridine ([(123)I]IaraU) was prepared in a radiochemical yield of 8% and specific activity of 21 GBq/µmol, respectively. Inhibition of the cellular uptake of these two tracers was compared by using the arabinosyl uridine analogs such as 5-iodo, 5-fluoro and 5-(E)-iodovinyl arabinosyl uridine along with 2'-fluoro-5-iodo arabinosyl uridine (FIAU). Due to potential instability of the iodo group, accumulation index of 1.6 for [(123)I]IaraU by HSV1-TK vs. control cells could virtually be achieved at 1.5 h, but dropped to 0.2 compared to 2.0 for [(18)F]FLT at 5 h. The results from competitive inhibition by these nucleosides against the accumulation of [(18)F]FLT implied that FLT exerted a mixed TK1- and TK2-dependent inhibition with HSV1-tk gene transfection because of the shifting of thymidine kinase status. Taken together, the combination of [(18)F]FLT and HSV1-TK provides a synergistic imaging potency.

Magnetically directed self-assembly of electrospun superparamagnetic fibrous bundles to form three-dimensional tissues with a highly ordered architecture.

Engineering three-dimensional (3D) cell-dense tissues with a well-organized structure remains a challenge in tissue engineering. In this study, highly oriented fibrous bundles, consisted of composite fibers of poly(L-lactide-co-glycolide)/superparamagnetic iron oxide nanoparticles, were fabricated using an electrospinning technique. The magnetic properties of the fabricated fibrous bundles were examined by a vibrating sample magnetometer and a superconducting quantum interference device; the results demonstrate that the fabricated fibrous bundles revealed superparamagnetic behavior without magnetic hysteresis. After seeding C2C12 myoblasts on the fibrous bundles, cells were grown along the direction of the underlying fibers (cell rods), an aligned pattern similar to those in native skeletal muscle tissues. When treated with the differentiation medium, myoblasts were fused together and formed multinucleated myotubes. As soon as applying an external magnetic field, the cell rods can spontaneously response to the magnetic control and self-assemble into 3D tissues with a highly ordered architecture. These findings demonstrate that the magnetically susceptible fibrous bundles not only can serve as a functional unit providing the topographic cue for cell orientation, but also can be magnetically manipulated for the creation of 3D cell-dense constructs. This technique may be applied to various cell types and scaffold configurations, thus advancing the design of engineered tissues that more closely replicate native tissues.

Type II collagen-chondroitin sulfate-hyaluronan scaffold cross-linked by genipin for cartilage tissue engineering.

Owing to of the limited repair capacity of articular cartilage, it is essential to develop tissue-engineered cartilage for patients suffering from joint disease. Chondroitin sulfate (CS) and hyaluronan (HA) are the components of the cartilage extracellular matrix (ECM) and are known to influence the proliferation and differentiation of chondrocytes. Scaffolds composed of type-II collagen, CS, and HA may create an environment that can preserve the normal phenotype of cells to promote regeneration of cartilage-like constructs. In this investigation, we prepared and characterized 3-dimensional type-II collagen scaffolds both with and without HA and CS. Porous composite scaffolds fabricated by freeze-drying showed interconnected pores with mean diameters of 140+/-30 microm and porosities of 92-95% after cross-linking with genipin. After a 14-day in vitro culture, morphologically round chondrocytes were found to be uniformly distributed throughout the sponges. Expression of genes of aggrecan, type-II collagen and cartilage oligomeric matrix protein (COMP) was statistically and significantly increased on scaffolds with CS and HA than those without CS and HA. Furthermore, there was a markedly greater accumulation of proteoglycans (PGs) on the scaffolds with CS and HA.

Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1.

Curcumin (diferuloylmethane) is an active component of the spice turmeric and has a diversity of antitumor activities. In this study, we found that curcumin can inhibit cancer cell invasion and metastasis through activation of the tumor suppressor DnaJ-like heat shock protein 40 (HLJ1). Human lung adenocarcinoma cells (CL1-5) treated with curcumin (1-20 mumol/L) showed a concentration-dependent reduction in cell migration, invasion, and metastatic ability, and this was associated with increased HLJ1 expression. Knockdown of HLJ1 expression by siRNA was able to reverse the curcumin-induced anti-invasive and antimetastasis effects in vitro and in vivo. The HLJ1 promoter and enhancer in a luciferase reporter assay revealed that curcumin transcriptionally up-regulates HLJ1 expression through an activator protein (AP-1) site within the HLJ1 enhancer. JunD, one of the AP-1 components, was significantly up-regulated by curcumin (1-20 mumol/L) in a concentration- and time-dependent manner. Knockdown of JunD expression could partially reduce the curcumin-induced HLJ1 activation and diminish the anti-invasive effect of curcumin, indicating that JunD would seem to be involved in curcumin-induced HLJ1 expression. Curcumin was able to induce c-Jun NH(2)-kinase (JNK) phosphorylation, whereas the JNK inhibitor (SP-600125) could attenuate curcumin-induced JunD and HLJ1 expression. Activation of HLJ1 by curcumin further leads to up-regulation of E-cadherin and a suppression of cancer cell invasion. Our results show that curcumin induces HLJ1, through activation of the JNK/JunD pathway, and inhibits lung cancer cell invasion and metastasis by modulating E-cadherin expression. This is a novel mechanism and supports the application of curcumin in anti-cancer metastasis therapy.

Synchronous advanced scrotal verrucous carcinoma with peripheral T-cell lymphoma.

Scrotal verrucous carcinoma is extremely rare. Classic cases have been suggested to be occupation related. We report on a 65-year-old male drifter, who had concomitant multiple condyloma acuminatum and advanced verrucous carcinoma of the right hemiscrotal skin with peripheral T-cell lymphoma and extensive lymphadenopathy. To our knowledge, it is the first case of synchronous verrucous carcinoma and peripheral T-cell lymphoma. The patient died of disseminated disease 2 months after undergoing wide excision of the tumor. A brief discussion of the diagnosis, treatment, and prognosis is presented.

Structure and function of the XpsE N-terminal domain, an essential component of the Xanthomonas campestris type II secretion system.

Secretion of fully folded extracellular proteins across the outer membrane of Gram-negative bacteria is mainly assisted by the ATP-dependent type II secretion system (T2SS). Depending on species, 12-15 proteins are usually required for the function of T2SS by forming a trans-envelope multiprotein secretion complex. Here we report crystal structures of an essential component of the Xanthomonas campestris T2SS, the 21-kDa N-terminal domain of cytosolic secretion ATPase XpsE (XpsEN), in two conformational states. By mediating interaction between XpsE and the cytoplasmic membrane protein XpsL, XpsEN anchors XpsE to the membrane-associated secretion complex to allow the coupling between ATP utilization and exoprotein secretion. The structure of XpsEN observed in crystal form P4(3)2(1)2 is composed of a 90-residue alpha/beta sandwich core domain capped by a 62-residue N-terminal helical region. The core domain exhibits structural similarity with the NifU-like domain, suggesting that XpsE(N) may be involved in the regulation of XpsE ATPase activity. Surprisingly, although a similar core domain structure was observed in crystal form I4(1)22, the N-terminal 36 residues of the helical region undergo a large structural rearrangement. Deletion analysis indicates that these residues are required for exoprotein secretion by mediating the XpsE/XpsL interaction. Site-directed mutagenesis study further suggests the more compact conformation observed in the P4(3)2(1)2 crystal likely represents the XpsL binding-competent state. Based on these findings, we speculate that XpsE might function in T2SS by cycling between two conformational states. As a closely related protein to XpsE, secretion ATPase PilB may function similarly in the type IV pilus assembly.