Enhanced Anti-Cancer Effects of Conditioned Medium from Hypoxic Human Adult Dermal Fibroblasts on Cervical Cancer Cells.

Hypoxia regulates fibroblast function by changing intracellular signaling and secretion factors, that influence the states of nearby cells. In this work, we investigated how medium (CM) from human adult dermal fibroblasts (HDFs) cultured in normoxic and hypoxic conditions affected cervical cancer (HeLa) cells. The HeLa cells showed decreased cell viability, increased apoptosis, and cell cycle arrest in response to CM from hypoxic-cultured HDFs (H-CM) compared with CM from normoxic-cultured HDFs (N-CM). Among the proteins up-regulated (>2-fold) in H-CM compared with N-CM, lymphotoxin-beta receptor (LTBR) decreased the viability of HeLa cells. Among the intracellular proteins down-regulated (>2-fold) in HeLa cells treated with H-CM compared with N-CM, the most enriched biological process GO term and KEGG pathway were protein deubiquitination and hsa05166:HTLV-I infection, respectively. In the protein−protein interaction network of intracellular proteins with altered expression (>2-fold), 1 up-regulated (TNF) and 8 down-regulated (ESR1, MCL1, TBP, CD19, LCK, PCNA, CHEK1, and POLA1) hub proteins were defined. Among the down-regulated hub proteins, the most enriched biological process GO term and KEGG pathway were leading strand elongation and hsa05166:HTLV-I infection, respectively. This study reveals that H-CM had stronger anti-cancer effects on cervical cancer cells than N-CM and induced intracellular signaling patterns related to those enhanced anti-cancer effects.

Sensitive optical bio-sensing of p-type WSe2 hybridized with fluorescent dye attached DNA by doping and de-doping effects.

Layered transition metal dichalcogenides, such as MoS2, WSe2 and WS2, are exciting two-dimensional (2D) materials because they possess tunable optical and electrical properties that depend on the number of layers. In this study, the nanoscale photoluminescence (PL) characteristics of the p-type WSe2 monolayer, and WSe2 layers hybridized with the fluorescent dye Cy3 attached to probe-DNA (Cy3/p-DNA), have been investigated as a function of the concentration of Cy3/DNA by using high-resolution laser confocal microscopy. With increasing concentration of Cy3/p-DNA, the measured PL intensity decreases and its peak is red-shifted, suggesting that the WSe2 layer has been p-type doped with Cy3/p-DNA. Then, the PL intensity of the WSe2/Cy3/p-DNA hybrid system increases and the peak is blue-shifted through hybridization with relatively small amounts of target-DNA (t-DNA) (50-100 nM). This effect originates from charge and energy transfer from the Cy3/DNA to the WSe2. For t-DNA detection, our systems using p-type WSe2 have the merit in terms of the increase of PL intensity. The p-type WSe2 monolayers can be a promising nanoscale 2D material for sensitive optical bio-sensing based on the doping and de-doping responses to biomaterials.

Enhancement of angiogenic effects by hypoxia-preconditioned human umbilical cord-derived mesenchymal stem cells in a mouse model of hindlimb ischemia.

It has been studied that mesenchymal stem cells (MSCs) have the capability to promote angiogenesis. Furthermore, there is strong evidence that hypoxic conditions can enhance angiogenesis and immune modulation mediated by MSCs, a notion that has been applied in many fields of clinical application. In the present study, we compared the efficacy of hypoxia preconditioned human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) and normoxia conditioned hUC-MSCs for the treatment of ischemic injury in hindlimbs of an immunodeficient mouse model. Expression of negative markers for MSC such as CD31, CD34, and CD45 or positive markers such as CD44, CD73, CD90, and CD105 was not significantly changed in hypoxia preconditioned hUC-MSCs compared with hUC-MSCs cultured in normoxic condition. Expression of angiogenesis-related genes such as COX-2, VEGF, Tie-2, and TGF-ß1 was increased compared with hUC-MSCs cultured in normoxic conditions. In the in vivo model, CD31 expression as a marker of angiogenesis was significantly increased in the ischemic limbs at 1 month after injection with hypoxic hUC-MSCs. Angiogenesis-related genes such as Ang-1, COX-1, PIGF, and MCP-1 were significantly upregulated in the muscle of ischemic hindlimbs treated with hypoxic hUC-MSCs than normoxic hUC-MSCs. Expression of proinflammatory genes such as IL-1, and IL-20 was reduced, whereas TGF-ß1, which has an anti-inflammatory effect, was strongly increased. In conclusion, hypoxic culture conditions could induce expression of angiogenesis related genes in hUC-MSCs, and hypoxia preconditioned hUC-MSCs showed enhancing effects by inducing angiogenesis and low inflammatory immune response compared with normoxic hUC-MSCs in the ischemia injured hindlimb of immunodeficient mice.

Effects of stimulating interleukin -2/anti- interleukin -2 antibody complexes on renal cell carcinoma.

BACKGROUND: Current therapies for advanced renal cell carcinoma (RCC) have low cure rates or significant side effects. It has been reported that complexes composed of interleukin (IL)-2 and stimulating anti-IL-2 antibody (IL-2C) suppress malignant melanoma growth. We investigated whether it could have similar effects on RCC. METHODS: A syngeneic RCC model was established by subcutaneously injecting RENCA cells into BALB/c mice, which were administered IL-2C or phosphate-buffered saline every other day for 4 weeks. RCC size was measured serially, and its weight was assessed 4 weeks after RENCA injection. Immune cell infiltration into RCC lesions and spleen was assessed by flow cytometry and immunohistochemistry. RESULTS: IL-2C treatment increased the numbers of CD8(+) memory T and natural killer (NK) cells in healthy BALB/c mice (P < 0.01). In the spleen of RCC mice, IL-2C treatment also increased the number of CD8(+) memory T, NK cells, and macrophages as compared to PBS-treated controls (P < 0.01). The number of interferon-γ- and IL-10-producing splenocytes increased and decreased, respectively after 4 weeks in the IL-2C-treated mice (P < 0.01). Tumor-infiltrating immune cells including CD4(+) T, CD8(+) T, NK cells as well as macrophages were increased in IL-2C-treated mice than controls (P < 0.05). Pulmonary edema, the most serious side effect of IL-2 therapy, was not exacerbated by IL-2C treatment. However, IL-2C had insignificant inhibitory effect on RCC growth (P = 0.1756). CONCLUSIONS: IL-2C enhanced immune response without significant side effects; however, this activity was not sufficient to inhibit RCC growth in a syngeneic, murine model.

IL-2/anti-IL-2 complex attenuates renal ischemia-reperfusion injury through expansion of regulatory T cells.

Regulatory T cells (Tregs) can suppress immunologic damage in renal ischemia-reperfusion injury (IRI), but the isolation and ex vivo expansion of these cells for clinical application remains challenging. Here, we investigated whether the IL-2/anti-IL-2 complex (IL-2C), a mediator of Treg expansion, can attenuate renal IRI in mice. IL-2C administered before bilateral renal IRI induced Treg expansion in both spleen and kidney, improved renal function, and attenuated histologic renal injury and apoptosis after IRI. Furthermore, IL-2C administration reduced the expression of inflammatory cytokines and attenuated the infiltration of neutrophils and macrophages in renal tissue. Depletion of Tregs with anti-CD25 antibodies abrogated the beneficial effects of IL-2C. However, IL-2C-mediated renal protection was not dependent on either IL-10 or TGF-ß. Notably, IL-2C administered after IRI also enhanced Treg expansion in spleen and kidney, increased tubular cell proliferation, improved renal function, and reduced renal fibrosis. In conclusion, these results indicate that IL-2C-induced Treg expansion attenuates acute renal damage and improves renal recovery in vivo, suggesting that IL-2C may be a therapeutic strategy for renal IRI.

In-Depth Analysis on Self Alignment Effect of the Fermi-Level Using Graphene on Both n- and p-Type Semiconductors.

Optimizing the contact structure while reducing the contact resistance in advanced transistors has become an extremely challenging problem. Because the existing techniques are limited to controlling only one semiconductor type, either n- or p-type, owing to their work function differences, significant challenges are encountered in the integration of a contact structure and metal suitable for both n- and p-type semiconductors. This is a formidable drawback of the complementary metal-oxide-semiconductor (CMOS) technology. In this paper, we demonstrate the effectiveness of a metal/graphene/semiconductor (MGrS) as a universal source/drain contact structure for both n- and p-type transistors. The MGrS contact structure significantly enhanced the reverse current density (JR) and reduced the Schottky barrier height (SBH) for both semiconductor types. From the analysis of the SBH values and their relationship with the metal work function, which refers to the S-parameter, the van der Waals contact of graphene (Gr) effectively alleviated the Fermi level (FL) pinning for both semiconductor types, reducing the metal-induced gap states (MIGS) at the Gr/semiconductor interface. Furthermore, Gr effectively modulated the work function of the contact metal to yield a position favorable for each semiconductor type. Consequently, a single MGrS contact structure on a Si substrate resulted in excellent Ohmic contacts in both n- and p-type Si, with SBH values reduced to 0.012 and 0.024 eV for n- and p-type Si, respectively. This new approach for integrating the contact structures of semiconductor types will lead to extended capabilities for high-performance device applications and CMOS logical circuitry.

Highly Tunable Negative Differential Resistance Device Based on Insulator-to-Metal Phase Transition of Vanadium Dioxide.

Negative differential resistance (NDR) based on the band-to-band tunneling (BTBT) mechanism has recently shown great potential in improving the performance of various electronic devices. However, the applicability of conventional BTBT-based NDR devices is restricted by their insufficient performance due to the limitations of the NDR mechanism. In this study, we develop an insulator-to-metal phase transition (IMT)-based NDR device that exploits the abrupt resistive switching of vanadium dioxide (VO2) to achieve a high peak-to-valley current ratio (PVCR) and peak current density (Jpeak) as well as controllable peak and valley voltages (Vpeak/valley). When a phase transition is induced in VO2, the effective voltage bias on the two-dimensional channel is decreased by the reduction in the VO2 resistance. Accordingly, the effective voltage adjustment induced by the IMT results in an abrupt NDR. This NDR mechanism based on the abrupt IMT results in a maximum PVCR of 71.1 through its gate voltage and VO2 threshold voltage tunability characteristics. Moreover, Vpeak/valley is easily modulated by controlling the length of VO2. In addition, a maximum Jpeak of 1.6 × 106 A/m2 is achieved through light-tunable characteristics. The proposed IMT-based NDR device is expected to contribute to the development of various NDR devices for next-generation electronics.

Ultralow Schottky Barrier Height Achieved by Using Molybdenum Disulfide/Dielectric Stack for Source/Drain Contact.

Energy barrier formed at a metal/semiconductor interface is a critical factor determining the performance of nanoelectronic devices. Although diverse methods for reducing the Schottky barrier height (SBH) via interface engineering have been developed, it is still difficult to achieve both an ultralow SBH and a low dependence on the contact metals. In this study, a novel structure, namely, a metal/transition-metal dichalcogenide (TMD) interlayer (IL)/dielectric IL/semiconductor (MTDS) structure, was developed to overcome these issues. Molybdenum disulfide (MoS2) is a promising TMD IL material owing to its interface characteristics, which yields a low SBH and reduces the reliance on contact metals. Moreover, an ultralow SBH is achieved via the insertion of an ultrathin ZnO layer between MoS2 and a semiconductor, thereby inducing an n-type doping effect on the MoS2 IL and forming an interface dipole in the favorable direction at the ZnO IL/semiconductor interfaces. Consequently, the lowest SBH (0.07 eV) and a remarkable improvement in the reverse current density (by a factor of approximately 5400) are achieved, with a wide room for contact-metal dependence. This study experimentally and theoretically validates the effect of the proposed MTDS structure, which can be a key technique for next-generation nanoelectronics.

Metabolites can regulate stem cell behavior through the STAT3/AKT pathway in a similar trend to that under hypoxic conditions.

Stem cell therapy has long been considered a promising mode of treatment for many incurable diseases. Human mesenchymal stem cells (hMSCs) have provided the most promising results to date for regenerative medicine. Nevertheless, due to several obstacles such as difficulty in sourcing and characterizing hMSCs, they remain largely unavailable for clinical use. The signaling requirements for maintaining stem cell function have been studied widely, but little is known about how metabolism contributes to stem cell function. hMSCs have been shown to promote therapeutic efficacy in hypoxic conditions through metabolic conversion. According to published studies, certain metabolites are able to convert stem cell metabolism from oxidative phosphorylation to glycolysis. In this study, we selected several metabolites (fructose-1,6-bisphosphate (FBP), Phosphoenolpyruvic acid (PEP) and sodium oxalate (OXA)) to examine the relation between metabolites and stem cell functions. In addition, we investigated the ability of selected metabolites to induce rapid expansion of this cell population. Our results indicate that selected metabolites stimulate stem cell proliferation by induce glycolytic metabolism via AKT/STAT signaling.

Reduction of Threshold Voltage Hysteresis of MoS2 Transistors with 3-Aminopropyltriethoxysilane Passivation and Its Application for Improved Synaptic Behavior.

Although molybdenum disulfide (MoS2) is highlighted as a promising channel material, MoS2-based field-effect transistors (FETs) have a large threshold voltage hysteresis (Δ VTH) from interface traps at their gate interfaces. In this work, the Δ VTH of MoS2 FETs is significantly reduced by inserting a 3-aminopropyltriethoxysilane (APTES) passivation layer at the MoS2/SiO2 gate interface owing to passivation of the interface traps. The Δ VTH is reduced from 23 to 10.8 V by inserting the 1%-APTES passivation layers because APTES passivation prevents trapping and detrapping of electrons, which are the major source of the Δ VTH. The reduction in the density of interface traps ( Dit) is confirmed by the improvement of the subthreshold swing (SS) after inserting the APTES layer. Furthermore, the improvement in the synaptic characteristics of the MoS2 FET through the APTES passivation is investigated. Both inhibitory and excitatory postsynaptic currents (PSC) are increased by 33% owing to the reduction in the Δ VTH and the n-type doping effect of the APTES layer; moreover, the linearity of PSC characteristics is significantly improved because the reduction in Δ VTH enables the synaptic operation to be over the threshold region, which is linear. The application of the APTES gate passivation technique to MoS2 FETs is promising for reliable and accurate synaptic applications in neuromorphic computing technology as well as for the next-generation complementary logic applications.

Schottky Barrier Height Modulation Using Interface Characteristics of MoS2 Interlayer for Contact Structure.

Schottky barrier height (SBH) engineering of contact structures is a primary challenge to achieve high performance in nanoelectronic and optoelectronic applications. Although SBH can be lowered through various Fermi-level (FL) unpinning techniques, such as a metal/interlayer/semiconductor (MIS) structure, the room for contact metal adoption is too narrow because the work function of contact metals should be near the conduction band edge (CBE) of the semiconductor to achieve low SBH. Here, we propose a novel structure, the metal/transition metal dichalcogenide/semiconductor structure, as a contact structure that can effectively lower the SBH with wide room for contact metal adoption. A perpendicularly integrated molybdenum disulfide (MoS2) interlayer effectively alleviates FL pinning by reducing metal-induced gap states at the MoS2/semiconductor interface. Additionally, it can induce strong FL pinning of contact metals near its CBE at the metal/MoS2 interface. The technique using FL pinning and unpinning at metal/MoS2/semiconductor interfaces is first introduced in the MIS scheme to allow the use of various contact metals. Consequently, significant reductions of the SBH from 0.48 to 0.12 eV for GaAs and from 0.56 to 0.10 eV for Ge are achieved with several different contact metals. This work significantly reduces the dependence on contact metals with lowest SBH and proposes a new way of overcoming current severe contact issues for future nanoelectronic and optoelectronic applications.

FGF-17 from Hypoxic Human Wharton's Jelly-Derived Mesenchymal Stem Cells Is Responsible for Maintenance of Cell Proliferation at Late Passages.

BACKGROUND AND OBJECTIVES: Although it is well known that hypoxic culture conditions enhance proliferation of human mesenchymal stem cells, the exact mechanism is not fully understood. In this study, we investigated the effect of fibroblast growth factor (FGF)-17 from hypoxic human Wharton's Jelly-derived mesenchymal stem cells (hWJ-MSCs) on cell proliferation at late passages. METHODS AND RESULTS: hWJ-MSCs were cultured in α-MEM medium supplemented with 10% fetal bovine serum (FBS) in normoxic (21% O2) and hypoxic (1% O2) conditions. Protein antibody array was performed to analyze secretory proteins in conditioned medium from normoxic and hypoxic hWJ-MSCs at passage 10. Cell proliferation of hypoxic hWJ-MSCs was increased compared with normoxic hWJ-MSCs from passage 7 to 10, and expression of secretory FGF-17 was highly increased in conditioned medium from hypoxic hWJ-MSCs at passage 10. Knockdown of FGF-17 in hypoxic and normoxic hWJ-MSCs decreased cell proliferation, whereas treatment of hypoxic and normoxic hWJ-MSCs with recombinant protein FGF-17 increased their proliferation. Signal transduction of FGF-17 in hypoxic and normoxic hWJ-MSCs involved the ERK1/2 pathway. Cell phenotypes were not changed under either condition. Differentiation-related genes adiponectin, Runx2, and chondroadherin were downregulated in normoxic hWJ-MSCs treated with rFGF-17, and upregulated by siFGF-17. Expression of alkaline phosphatase (ALP), Runx2, and chondroadherin was upregulated in hypoxic hWJ-MSCs, and this effect was rescued by transfection with siFGF-17. Only chondroadherin was upregulated in hypoxic hWJ-MSCs with rFGF-17. CONCLUSIONS: In hypoxic culture conditions, FGF-17 from hypoxic hWJ-MSCs contributes to the maintenance of high proliferation at late passages through the ERK1/2 pathway.

Enhanced Anti-Cancer Effects of Conditioned Medium from Hypoxic Human Umbilical Cord-Derived Mesenchymal Stem Cells.

BACKGROUND AND OBJECTIVES: There have been contradictory reports on the pro-cancer or anti-cancer effects of mesenchymal stem cells. In this study, we investigated whether conditioned medium (CM) from hypoxic human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) (H-CM) showed enhanced anti-cancer effects compared with CM from normoxic hUC-MSCs (N-CM). METHODS AND RESULTS: Compared with N-CM, H-CM not only strongly reduced cell viability and increased apoptosis of human cervical cancer cells (HeLa cells), but also increased caspase-3/7 activity, decreased mitochondrial membrane potential (MMP), and induced cell cycle arrest. In contrast, cell viability, apoptosis, MMP, and cell cycle of human dermal fibroblast (hDFs) were not significantly changed by either CM whereas caspase-3/7 activity was decreased by H-CM. Protein antibody array showed that activin A, Beta IG-H3, TIMP-2, RET, and IGFBP-3 were upregulated in H-CM compared with N-CM. Intracellular proteins that were upregulated by H-CM in HeLa cells were represented by apoptosis and cell cycle arrest terms of biological processes of Gene Ontology (GO), and by cell cycle of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. In hDFs, negative regulation of apoptosis in biological process of GO and PI3K-Akt signaling pathway of KEGG pathways were represented. CONCLUSIONS: H-CM showed enhanced anti-cancer effects on HeLa cells but did not influence cell viability or apoptosis of hDFs and these different effects were supported by profiling of secretory proteins in both kinds of CM and intracellular signaling of HeLa cells and hDFs.

Schottky Barrier Height Engineering for Electrical Contacts of Multilayered MoS2 Transistors with Reduction of Metal-Induced Gap States.

The difficulty in Schottky barrier height (SBH) control arising from Fermi-level pinning (FLP) at electrical contacts is a bottleneck in designing high-performance nanoscale electronics and optoelectronics based on molybdenum disulfide (MoS2). For electrical contacts of multilayered MoS2, the Fermi level on the metal side is strongly pinned near the conduction-band edge of MoS2, which makes most MoS2-channel field-effect transistors (MoS2 FETs) exhibit n-type transfer characteristics regardless of their source/drain (S/D) contact metals. In this work, SBH engineering is conducted to control the SBH of electrical top contacts of multilayered MoS2 by introducing a metal-interlayer-semiconductor (MIS) structure which induces the Fermi-level unpinning by a reduction of metal-induced gap states (MIGS). An ultrathin titanium dioxide (TiO2) interlayer is inserted between the metal contact and the multilayered MoS2 to alleviate FLP and tune the SBH at the S/D contacts of multilayered MoS2 FETs. A significant alleviation of FLP is demonstrated as MIS structures with 1 nm thick TiO2 interlayers are introduced into the S/D contacts. Consequently, the pinning factor ( S) increases from 0.02 for metal-semiconductor (MS) contacts to 0.24 for MIS contacts, and the controllable SBH range is widened from 37 meV (50-87 meV) to 344 meV (107-451 meV). Furthermore, the Fermi-level unpinning effect is reinforced as the interlayer becomes thicker. This work widens the scope for modifying electrical characteristics of contacts by providing a platform to control the SBH through a simple process as well as understanding of the FLP at the electrical top contacts of multilayered MoS2.

Simple measurement of spinal cord evoked potential: a valuable data source in the rat spinal cord injury model.

Measurement of spinal cord evoked potentials (SCEPs) is proposed as a means of predicting locomotion outcome in the rat spinal cord injury (SCI) model. Using 55 rats, three reproducible peak waves (waves I, II and III) were observed during stimulation at the C7 level with recording at the L1 epidural space. Hemisection at the T13 level showed three wave loss patterns: wave III loss only, loss of both wave II and III, and loss of all three waves. Defining an ideal SCI model as establishment of stable monoparesis or paraparesis, all animals in the wave II-III loss group showed favorable results. Histological data and electrophysiological properties allowed reasonable assumptions of wave origin: wave I from extrapyramidal tracts, wave II from the ventral corticospinal tract, and wave III from the dorsal corticospinal tract. Complete destruction of pyramidal tracts in both dorsal and ventral fibers was essential for long-term impairment of locomotion.

Heme Oxygenase-1 Induction and Anti-inflammatory Actions of Atractylodes macrocephala and Taraxacum herba Extracts Prevented Colitis and Was More Effective than Sulfasalazine in Preventing Relapse.

BACKGROUND/AIMS: In inflammatory bowel disease (IBD), repeated bouts of remission and relapse occur in patients and can impose a risk of colitis-associated cancer. We hypothesized that plant extracts of Atractylodes macrocephala (AM) or Taraxacum herba (TH) may be better than sulfasalazine for treating this disease because these extracts can promote additional regeneration. METHODS: Murine intestinal epithelial IEC-6 cells were pretreated with AM or TH before a lipopolysaccharide (LPS)-induced challenge. Acute colitis was induced with 7 days of dextran sulfate sodium (DSS) in male C57BL/6 mice, and extracts of AM and TH were administered for 2 weeks before DSS administration. RESULTS: In vitro studies demonstrated that AM or TH treatment reduced LPS-induced COX-2 and tumor necrosis factor-α mRNA levels but increased heme oxygenase-1 (HO-1). Oral preadministration of AM and TH rescued mice from DSS-induced colitis by inhibiting inflammatory mediators via inactivated extracellular signal regulated kinase and repressed nuclear factor κB and signal transducer and activator of transcription 3, but the effect was weaker for sulfasalazine than that for the extracts. Anti-inflammatory activities occurred via the inhibition of macrophage and T lymphocyte infiltrations. Unlike sulfasalazine, which did not induce HO-1, TH extracts afforded significant HO-1 induction. CONCLUSIONS: Because the AM or TH extracts were far superior in preventing DSS-induced colitis than sulfasalazine, AM or TH extracts can be considered natural agents that can prevent IBD relapse.

Protein profiling and angiogenic effect of hypoxia-cultured human umbilical cord blood-derived mesenchymal stem cells in hindlimb ischemia.

The aim of the present study was to investigate protein profiles of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) cultured in normoxic (21% O2) and hypoxic (1% O2) conditions, and evaluate oxygenation effects on angiogenesis in an ischemic hindlimb mouse model using a modified ischemic scoring system. Hypoxic conditions did not change the expression of phenotypic markers and increased adipogenesis and chondrogenesis. Epidermal growth factor (EGF), transforming growth factor alpha (TGF-α), TGF-ß RII, and vascular endothelial growth factor (VEGF) were upregulated in the conditioned medium of hypoxic hUCB-MSCs, which are commonly related to angiogenesis and proliferation of biological processes by Gene Ontology. In the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, significant enrichment of the phosphorylation of abelson murine leukemia viral oncogene homolog 1 (ABL1) (Phospho-Tyr204) and B-cell lymphoma-extra large (BCL-XL) (Phospho-Thr47) as anti-apoptotic pathways was observed in hypoxic hUCB-MSCs. Furthermore, hypoxic conditions induced proliferation and migration, and reduced apoptosis of hUCB-MSCs in vitro. Based on the results of protein antibody array, we evaluated the angiogenic effects of injecting normoxic or hypoxic hUCB-MSCs (1×106) into the ischemic hindlimb muscles of mice. Ischemic scores and capillary generation were significantly greater in the hypoxic hUCB-MSC injection group than in the normoxic hUCB-MSC group. Our findings demonstrate that culturing hUCB-MSCs in hypoxic conditions not only significantly enriches phosphorylation in the anti-apoptosis pathway and enhances the secretion of several angiogenic proteins from cells, but also alleviates ischemic injury of hindlimb of mice.

Therapeutic Potential of Human Mesenchymal Stem Cells for Treating Ischemic Limb Diseases.

Ischemic limb diseases are induced by different obstructions of peripheral arteries. These obstructions result in insufficient nutrient and oxygen supplies to the extremities, thereby leading to severe tissue damage that is in turn related to severe morbidities and mortalities. Mesenchymal stem cells (MSCs) have been isolated from various sources. These cells are multipotent with respect to differentiation and are also characterized by migration, immune suppression, and secretion of paracrine factors. Mesenchymal stem cells have been proposed to have therapeutic potential for the treatment of ischemic limb diseases. In preclinical experiments, injection of single MSCs has been shown to increase angiogenesis and blood flow in ischemic hindlimb animal models; several molecular mechanisms of angiogenesis have also been elucidated. Furthermore, modified strategies have been developed for enhancing angiogenesis and the efficacy of MSCs. These strategies have demonstrated significant effects in pre-clinical studies. In clinical trials, MSCs have shown significant effects in the treatment of ischemic limb diseases. In this review, we focus on the therapeutic properties of human MSCs and the modified methods for enhancing angiogenesis in pre-clinical experiments. We also discuss the clinical applications of MSCs for treating limb ischemia.

Mechanism underlying cytotoxicity of thialysine, lysine analog, toward human acute leukemia Jurkat T cells.

We first report the mechanism for the inhibitory effect of the lysine analog, thialysine on human acute leukemia Jurkat T cells. When Jurkat T cells were treated with thialysine (0.32-2.5 mM), apoptotic cell death along with several biochemical events such as mitochondrial cytochrome c release, caspase-9 activation, caspase-3 activation, degradation of poly (ADP-ribose) polymerase, and DNA fragmentation was induced in a dose- and time-dependent manner. However, these thialysine-induced apoptotic events were significantly abrogated by an ectopic expression of Bcl-xL, which is known to block mitochondrial cytochrome c release. Decylubiquinone, a mitochondrial permeability transition pore inhibitor, also suppressed thialysine-induced apoptotic events. Comparison of the thialysine-induced alterations in the cell cycle distribution between Jurkat T cells transfected with Bcl-xL gene (J/Bcl-xL) and Jurkat T cells transfected with vector (J/Neo) revealed that the apoptotic cells were mainly derived from the cells accumulated in S and G2/M phases following thialysine treatment. The interruption of cell cycle progression in the presence of thialysine was accompanied by a significant decline in the protein level of cdk4, cdk6, cdc2, cyclin A, cyclin B1, and cyclin E. These results demonstrate that the cytotoxic activity of thialysine toward Jurkat T cells is attributable to not only apoptotic cell death mediated by a mitochondria-dependent death signaling pathway, but also interruption of cell cycle progression by a massive down-regulation in the level of cdks and cyclins.

A case of invasive pulmonary aspergillosis with direct invasion of the mediastinum and the left atrium in an immunocompetent patient.

We report a case of invasive pulmonary aspergillosis invading the mediastinum and the left atrium. A 70-year-old woman was hospitalized for dyspnea. She had been well controlled for her diabetes mellitus and hypertension. The chest X-ray disclosed mediastinal widening, and the computed tomography scan of the chest showed that there was a large mediastinal mass and this lesion extended into the left atrium and right bronchus. The cardiac echocardiography showed that a huge mediastinal cystic mass compressed in the right atrium and a hyperechoic polypoid lesion in the left. The pathology from the bronchoscopic biopsy observed abundant fungal hyphae which was stained with periodic acid-Schiff and Gomori's methenamine silver. Despite the treatment with antifungal agents, she died from cardiac tamponade after three months. Invasive pulmonary aspergillosis, which involves the mediastinum and the heart, is very rare in immunocompetent patients.