Asperpyrone F, a new dimeric naphtho-γ-pyrone from the edible fungus Pleurotus ostreatus.

A new dimeric naphtho-γ-pyrone, asperpyrone F (1), along with six known ones, asperpyrones B (2) and C (3), fonsecinones A (4) and B (5), aurasperones A (6) and E (7), have been isolated from the solid culture of the edible fungus Pleurotus ostreatus. The structures of 1-7 were determined mainly by NMR and MS experiments. The absolute configuration of compound 1 was assigned via the circular dichroism (CD) data analysis. Compounds 1-7 showed modest antioxidant and immunomodulatory activities. All compounds were isolated from the fungus P. ostreatus for the first time.

Capsaicin and dihydrocapsaicin induce apoptosis in human glioma cells via ROS and Ca2+­mediated mitochondrial pathway.

Human glioma is the most common type of primary brain tumor and one of the most invasive and aggressive tumors, which, even with treatments including surgery, radiotherapy and chemotherapy, often relapses and exhibits resistance to conventional treatment methods. Developing novel strategies to control human glioma is, therefore, an important research focus. The present study investigated the mechanism of apoptosis induction in U251 human glioma cells by capsaicin (Cap) and dihydrocapsaicin (DHC), the major pungent ingredients of red chili pepper, using the Cell Counting Kit­8 assay, transmission electron microscopy analysis, flow cytometry analysis, laser scanning confocal microscope analysis and immunohistochemical staining. Treatment of U251 glioma cells with Cap and DHC resulted in a dose­ and time­dependent inhibition of cell viability and induction of apoptosis, whereas few effects were observed on the viability of L929 normal murine fibroblast cells. The apoptosis­inducing effects of Cap and DHC in U251 cells were associated with the generation of reactive oxygen species, increased Ca2+ concentrations, mitochondrial depolarization, release of cytochrome c into the cytosol and activation of caspase­9 and ­3. These effects were further confirmed by observations of the anti­tumor effects of Cap and DHC in vivo in a U251 cell murine tumor xenograft model. These results demonstrate that Cap and DHC are effective inhibitors of in vitro and in vivo survival of human glioma cells, and provide the rationale for further clinical investigation of Cap and DHC as treatments for human glioma.

Protein tyrosine phosphatase 1B (PTP1B) is dispensable for IgE-mediated cutaneous reaction in vivo.

Mast cells play a critical role in allergic reactions. The cross-linking of FcεRI-bound IgE with multivalent antigen initiates a cascade of signaling events leading to mast cell activation. It has been well-recognized that cross linking of FcεRI mediates tyrosine phosphorylation. However, the mechanism involved in tyrosine dephosphorylation in mast cells is less clear. Here we demonstrated that protein tyrosine phosphatase 1B (PTP1B)-deficient mast cells showed increased IgE-mediated phosphorylation of the signal transducer and activator of transcription 5 (STAT5) and enhanced production of CCL9 (MIP-1γ) and IL-6 in IgE-mediated mast cells activation in vitro. However, IgE-mediated calcium mobilization, ß-hexaosaminidase release (degranulation), and phosphorylation of IκB and MAP kinases were not affected by PTP1B deficiency. Furthermore, PTP1B deficient mice showed normal IgE-dependent passive cutaneous anaphylaxis and late phase cutaneous reactions in vivo. Thus, PTP1B specifically regulates IgE-mediated STAT5 pathway, but is redundant in influencing mast cell function in vivo.

Pu-erh tea powder preventive effects on cisplatin-induced liver oxidative damage in Wistar rats.

BACKGROUND: Chemotherapy is one of the major means for control of malignancies, with cisplatin (CDDP) as one of the main agents, widely used for the treatment of various malignant solid tumors. However, prevention of hepatotoxicity from cisplatin is one of the urgent issues in cancer chemotherapy. In this study, we aimed to investigate the effects of pu-erh tea on hepatotoxicity through body weight and tissue antioxidant parameters like, liver coefficient, serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde(MDA) and glutathione (GSH) levels, and light microscopic evaluation by histological findings. MATERIALS AND METHODS: The rats were randomly divided into five groups: Control (n=10), cisplatin (3 mg/kg p.i., n=10), cisplatin+pu-erh (0.32 g/kg/day i.g., n=10), cisplatin+pu-erh (0.8 g/kg/day i.g., n=10) and cisplatin+pu-erh (1.6 g/kg/day i.g., n=10). Pu-erh tea powder was administrated for 31 consecutive days. The rats were sacrificed at the end on the second day after a single dose of cisplatin treatment for measuring indices. RESULTS: Pu-erh tea powder exhibited a protective effect by decreasing MDA and GSH and increasing the SOD and GSH-PX levels and GSH-PX/MDA ratio in comparison with the control group. Besides, pu-erh tea was also able to alleviate the pathological damage to some extent. CONCLUSION: Pu-erh tea powder is protective against cisplatin-induced liver oxidative damages, especially at the medium dosage (0.8 g/kg/d).

In vitro study on the feasibility of magnetic stent hyperthermia for the treatment of cardiovascular restenosis.

Thermal treatment or hyperthermia has received considerable attention in recent years due to its high efficiency, safety and relatively few side-effects. In this study, we investigated whether it was possible to utilize targeted thermal or instent thermal treatments for the treatment of restenosis following percutaneous transluminal coronary angioplasty (PTCA) through magnetic stent hyperthermia (MSH). A 316L stainless steel stent and rabbit vascular smooth muscle cells (VSMCs) were used in the present study, in which the inductive heating characteristics of the stent under alternative magnetic field (AMF) exposure, as well as the effect of MSH on the proliferation, apoptosis, cell cycle and proliferating cell nuclear antigen (PCNA) expression of the rabbit VSMCs, were evaluated. The results demonstrated that 316L stainless steel coronary stents possess ideal inductive heating characteristics under 300 kHz AMF exposure. The heating properties were shown to be affected by the field intensity of the AMF, as well as the orientation the stent axis. MSH had a significant effect on the proliferation and apoptosis of VSMCs, and the effect was temperature-dependent. While a mild temperature of 43°C demonstrated negligible effects on the growth of VSMCs, MSH treatment above 47°C effectively inhibited the VSMC proliferation and induced apoptosis. Furthermore, a 47°C treatment exhibited a significant and long-term inhibitory effect on VSMC migration. The results strongly suggested that MSH may be potentially applied in the clinic as an alternative approach for the prevention and treatment of restenosis.

Antitumor effect and immune response induced by local hyperthermia in B16 murine melanoma: Effect of thermal dose.

This study aimed at investigating the antitumor effect and immune response induced by local high-temperature hyperthermia at different thermal doses in B16 murine melanoma. The screened optimal thermal dose (50°C, 15 min) which was demonstrated to be the most effective in immune response activation was applied to the treatment of lung metastasis. The optimal thermal dose was determined by evaluating the tumor volume change, survival period of tumor-bearing mice, and immune indices including interleukin (IL)-2, interferon (IFN)-γ and TNF-α mRNA expression in the spleen of mice subjected to local hyperthermia at various thermal doses. The activation of the immune response was further investigated by rechallenging the cured mice 60 days after hyperthermia treatment. The screened optimal thermal dose combined with immunoadjuvant compound 48/80 was applied for melanoma lung metastasis. While local hyperthermia effectively inhibited B16 melanoma tumor growth and prolonged the survival period of tumor-bearing mice, the antitumor immunity was significantly enhanced and the effect was thermal dose-dependent. Higher temperatures (≥50°C) induced a significant effect even with a short treatment time (≤15 min). No tumor regrowth was observed for rechallenged B16 melanoma in mice following treatment with local hyperthermia at a higher temperature. Local hyperthermia by optimal thermal dose in combination with immunoadjuvant compound 48/80 is an effective approach for the treatment of B16 melanoma lung metastasis. This study indicated that the use of a local high-temperature hyperthermia protocol inhibits tumor growth and stimulates a favorable antitumor immune response against malignant melanoma. The results of these experiments may have clinical significance for the treatment of melanoma.

Magnetic fluid hyperthermia induced by radiofrequency capacitive field for the treatment of transplanted subcutaneous tumors in rats.

Magnetic fluid hyperthermia (MFH) induced by a magnetic field has become a new heating technology for the treatment of malignant tumors due to its ability to heat the tumor tissue precisely and properly, and due to its significant therapeutic effects. In this study, MFH induced by radiofrequency capacitive field (RCF) for the treatment of transplanted subcutaneous tumors in rats, was investigated. A total of 50 rats bearing subcutaneous tumors were randomly divided into five groups, including i) a pseudo-treatment (PT) control group, ii) magnetic fluid (MF) group, iii) pure hyperthermia (PH) group, iv) magnetic fluid hyperthermia 1 (MFH1) group, and v) magnetic fluid hyperthermia 2 (MFH2) group. Tumors were irradiated for 30 min in the MFH1 group 24 h following injection of MF. Tumors were irradiated for 30 min in the MFH2 group 24 h following injection of MF, and irradiation was repeated for 30 min 72 h following injection of MF. Tumor volumes, tumor volume inhibition ratios and survival times in the rat model were examined. Temperatures of tumor cores and rims both rapidly reached the desired temperature (∼50°C) for tumor treatment within 5 to 10 min in the MFH1 and MFH2 groups, and we maintained this temperature level by manually adjusting the output power (70-130 W). Tumor volumes of the MFH1 and MFH2 groups were reduced compared to those of the PT, MF and PH groups. The inhibitory effect on tumor growth in the MFH2 group (91.57%) was higher compared to that in the MFH1 group (85.21%) and the other groups. The survival time of the MFH2 group (51.62±2.28 days) and MFH1 group (43.10±1.57 days) was increased compared to that of the PH, MF and PT groups. The results obtained show that MFH induced by RCF may serve as a potential and promising method for the treatment of tumors.

Magnetic stent hyperthermia for esophageal cancer: an in vitro investigation in the ECA-109 cell line.

Magnetic stent hyperthermia (MSH) is a novel approach for targeted thermotherapy for esophageal cancer, which is based on the mechanism that inductive heat can be generated by the esophageal stent upon exposure under an alternative magnetic field (AMF). A positive effect of MSH on esophageal cancer has been demonstrated, however, there is no study on the in vitro effects of heating treatment or of the effects of AMF exposure on human esophageal cancer cells. This study aimed to investigate the effect of MSH and of AMF exposure in esophageal cancer cells. Inductive heating characteristics of esophageal stents were assessed by exposing the stents under AMF. A rather rapid temperature rise of the Ni-Ti stent when subjected to AMF exposure was observed and the desired hyperthermic temperature could be controlled by adjusting the field parameter of the AMF. Human esophageal squamous carcinoma (ESCC) ECA-109 cells were divided into four groups: the control group, the water-bath heating group, the MSH group and the AMF exposure group. Hyperthermic temperatures were 43, 48 and 53˚C and the treatment time was in the range of 5-30 min. The MTT assay, apoptotic analysis and TUNEL staining were applied in the current investigation. Exposure of ECA-109 cells under AMF with a field intensity of 50 to 110 kA/m had negligible effect on cell viability, cell necrosis and apoptosis. Hyperthermia had a remarkable inhibitory effect on the cell viability and the effect was dependent on the thermal dose (temperature and time). The optimal thermal dose of MSH for ECA-109 cells was 48˚C for 20-30 min. The study also elucidated that there was a difference in the effects on cell necrosis and apoptosis between the heating mode of water bath and MSH. The data suggest that MSH may have clinical significance for esophageal cancer treatment.

Liquiritigenin attenuates the learning and memory deficits in an amyloid protein precursor transgenic mouse model and the underlying mechanisms.

The present paper is to examine whether liquiritigenin is able to attenuate the Alzheimer's-like learning and memory deficits in a transgenic (Tg) mouse model that over-expresses amyloid protein precursor (APP), and explores the underlying mechanisms. Consistent with our previous observations, we found that treatment with liquiritigenin improved the behavioral performance of Tg mice and it attenuated the protein expression of oligomeric form of amyloid ß-peptide (Aß). Furthermore, treatment with liquiritigenin inhibited astrocytosis in the hippocampus, and it may through its inhibitory activities on Notch-2, an important molecular regulating neural proliferation and differentiation. These findings provide evidence for beneficial activity of liquiritigenin in a mouse model of Alzheimer's disease and support the continued investigation of Notch signaling pathway as a target for treatment of Alzheimer's disease.

Regulator of calcineurin 1 (Rcan1) is required for the development of pulmonary eosinophilia in allergic inflammation in mice.

The presence of eosinophils in the lung is often regarded as a defining feature of asthma. On allergen stimulation, numbers of eosinophils and their progenitors are increased in both the bone marrow and lungs. Eosinophil progenitors provide an ongoing supply of mature eosinophils. Here, we report that deficiency in the regulator of calcineurin 1 gene (Rcan1) leads to a near-complete absence of eosinophilia in ovalbumin-induced allergic asthma in mice. In the absence of Rcan1, bone marrow cells produce significantly fewer eosinophils in vivo and in vitro on interleukin-5 stimulation. Importantly, eosinophil progenitor populations are significantly reduced in both naïve and ovalbumin-challenged Rcan1(-/-) mice. Bone marrow cells from Rcan1(-/-) mice are capable of developing into fully mature eosinophils, suggesting that Rcan1 is required for eosinophil progenitor production but may not be necessary for eosinophil maturation. Thus, Rcan1 represents a novel contributor in the development of eosinophilia in allergic asthma through regulation of eosinophil progenitor production.

Feasibility study of high-temperature thermoseed inductive hyperthermia in melanoma treatment.

Current treatment modalities for melanoma do not offer satisfactory efficacy. We have developed a new, minimally invasive hyperthermia technology based on radio-frequency hyperthermia. Herein, we investigated the feasibility of using a nickel-copper thermoseed for inductive hyperthermia at a relatively high temperature (46-55 ˚C). In vitro, the thermoseed showed good thermal effects and effective killing of B16/F10 melanoma cells. Temperatures of 53.1 ± 0.5 ˚C were achieved for a single thermoseed and 56.5 ± 0.5 ˚C for two in parallel (spacing 5 mm). No B16/F10 melanoma cells survived with heating time longer than 20 min in the parallel thermoseed group. Magnetic fields or thermoseeds alone did not affect the survival rate of B16/F10 cells (P>0.05). In vivo, B16/F10 melanoma cells were subcutaneously injected into the right axilla of C57BL/6 mice. After the tumors grew to ~11-13 mm, two thermoseeds (spacing 5 mm) were implanted into the tumors and the mice were subjected to an alternating magnetic field (100-250 kHz, 15 kA/m) to induce hyperthermia. The temperature at the center of the tumor reached 46 ˚C at 5 min and plateaued at 50 ˚C. Thermoseed treatment produced large necrotic areas, inhibited tumor growth in 60% (6 of 10) of animals and prolonged survival time (P<0.05). Thus, with further optimization and testing, high-temperature thermoseed inductive hyperthermia may have therapeutic potential for melanoma.

Apoptotic effects of Tian-Long compound on endometrial adenocarcinoma cells in vitro.

The Tian-Long (TL) compound is a water-soluble extract of six Chinese medicinal herbs. To explore its antitumor properties and the mechanism for activity in gynecological malignancies, the present studies were carried out using Ishikawa cells derived from uterine endometrial adenocarcinoma. Morphologically, cell death and decrease in the number of viable cells were observed in the presence of the TL compound. The proliferation of Ishikawa cells was significantly suppressed in a time- and dose-dependent manner, as indicated by both the WST-1 and the BrdU incorporation assay. Results from both the WST-1 and the BrdU incorporation assay demonstrated that the compound could inhibit the cell proliferation despite the presence of 17beta-estradiol in the medium. It is generally noted that the disturbance in mitochondrial function and DNA synthesis during cell proliferation can result in apoptosis. Being consistent with this notion, redistribution of the plasma membrane phosphatidylserine was identified with fluoromicroscopy and flow cytometry. Analysis of the fluorescent patterns of JC-1 staining revealed depolarization of the mitochondrial membrane in the exposed cells. Moreover, the amount of Bcl-2 enhanced in the presence of 17beta-estradiol was repressed by the compound. The present results indicate that the ingredients of TL compound are very promising for use in the treatment of endometrial adenocarcinoma. Further studies are necessary to elucidate the therapeutic mechanisms in its antitumor activity.

[Experimental study of Solanum nigrum on inhibiting angiogenesis in chick chorioallantoic membrane].

OBJECTIVE: To study the inhibitory effect of Solanum nigrum on angiogenesis. METHOD: We examined the effects of S. nigrum on angiogenesis in the chick chorioallantoic membrane (CAM) model. On day 7 of chick embryo incubation, three concentrations of S. nigrum aqueous extracts were applied to CAMs, and their effects were evaluated on day 9. RESULT: The angiogenesis area was significantly smaller in the CAM treated with S. nigrum than that in the control group (P < 0.001). Pathology analysis indicated that less angiogenesis occurred in the tissue of CAM under the filter paper treated with S. nigrum and the structure of large arteries was destroyed. The surrounding CAM showed a few angiogenesis formation. However, in the control group, a number of angiogenesis were observed. CONCLUSION: S. nigrum could inhibit the angiogenesis on CAM.

The early growth response factor-1 contributes to interleukin-13 production by mast cells in response to stem cell factor stimulation.

Stem cell factor (SCF) is not only critical for mast cell development, but also an important mast cell functional regulator. However, roles of transcription factors involved in SCF-induced effects remain incompletely defined. Early growth response factor-1 (Egr-1) is a member of zinc-finger transcription factor family. Mouse bone marrow-derived mast cells (BMMC) were used to examine a role of Egr-1 in SCF-induced mast cell activation and growth. SCF induced a strong and rapid expression of Egr-1 mRNA as tested by real-time PCR analysis. SCF-induced Egr-1 nuclear translocation and DNA binding were demonstrated by electrophoretic mobility shift assay (EMSA) and immunofluorescence assay. To examine if Egr-1 is required for SCF-induced IL-13 expression, Egr-1-deficient BMMC were used. Levels of SCF-induced IL-13 mRNA and protein were reduced in Egr-1 deficient BMMC when compared with wild-type BMMC. Although Egr-1 is required for macrophage and lymphocyte development, SCF-induced mast cells growth was not affected by Egr-1 deficiency. Interestingly, SCF-induced Egr activation was blocked by a tyrosine kinase inhibitor PP2, suggesting a role of tyrosine phosphorylation in SCF-induced Egr-1 activation. Taken together, our results suggest that Egr-1 is required for SCF-induced IL-13 expression, but not mast cell growth.

[Diffraction enhanced imaging of condylar cartilage of osteoarthritic rats].

OBJECTIVE: To observe the images of early lesions of condylar cartilage of osteoarthritic rats in synchrotron radiation diffraction enhanced imaging (DEI). METHODS: The animal model of temporomandibular joint osteoarthrosis was established in rat following the method of partial resection of the joint disc. The changes of osteoarthritic condylar cartilage in different pathological stages were observed by DEI and compared with those in situ histopathological sections. RESULTS: With DEI, straight and orbicular lines were detected in condylar cartilage 45 to 60 days after discs resection. The lines were confirmed by histopathology to be collagen degradation and tiny fissure formation inside the cartilage. CONCLUSIONS: DEI is capable of imaging the early stages of pathological changes of excised condylar cartilage such as collagen degradation and tiny fissure formation, and this technique is of potential value to clinical application.

The early growth response factor-1 is involved in stem cell factor (SCF)-induced interleukin 13 production by mast cells, but is dispensable for SCF-dependent mast cell growth.

The stem cell factor (SCF) plays a central role in the regulation of mast cell function and growth. However, roles of transcription factors involved in these processes remain incompletely defined. The early growth response factor-1 (Egr-1) is a member of the zinc finger transcription factor family. A role for Egr-1 in SCF-induced mast cell activation and growth was investigated in mouse bone marrow-derived mast cells (BMMC). The stimulation of BMMC with SCF induced a strong expression of Egr-1 mRNA. SCF-induced Egr-1 nuclear translocation and DNA binding were demonstrated by electrophoretic mobility shift assay (EMSA) and immunofluorescence assay. SCF-induced IL-13 expression was significantly reduced at both mRNA and protein levels in Egr-1-deficient BMMC. In addition, the synergy between IgE and SCF on IL-13 and IL-4 production was reduced in Egr-1-deficient mast cells. Interestingly, Egr-1 deficiency had little effect on SCF-induced mast cell growth. SCF-induced Egr activation likely requires tyrosine phosphorylation because a tyrosine kinase inhibitor PP2 blocked SCF-induced nuclear protein binding to Egr probe as determined by EMSA. Thus, Egr-1 is required for SCF-induced IL-13 expression, but not mast cell growth. Egr-1 represents a novel mechanism for SCF-induced mast cell activation.

[Review about mechanisms of anti-cancer of Solanum nigrum].

This paper gave a brief introduction of the effect of Solanum nigrum on anti-cancer. The experimental results showed that the total alkaloid isolated from S. nigrum interfered structure and function of tumor cell membrane, disturbed the synthesis of DNA and RNA, changed the cell cycle distribution, so that total alkaloids could play in inhibabition to tumor cells, while the glycoprotein (150 x 10(3)) isolated from S. nigrum might have shown anti-cancer abilities by blocking the anti-apoptotic pathway of NF-kappaB, activating caspase cascades reaction and increasing the production of nitric oxide.

[Development of measurement and control technologies for hyperthermia treatments of tumors with AC magnetic field].

Hyperthermia therapy with AC magnetic field heating magnetic nanoparticles is a new kind of treatment method. The paper reviews the research progresses about AC magnetic heating setups for hyperthermia therapy measurements of magnetic field temperature control, and so on.

Inductive heat property of Fe3O4/polymer composite nanoparticles in an ac magnetic field for localized hyperthermia.

The magnetite (Fe(3)O(4)) nanoparticles were prepared by coprecipitation of Fe(3+) and Fe(2+) with an aqueous NaOH solution. The Fe(3)O(4)/polyaniline (PANI) magnetic composite nanoparticles with a core-shell structure with a diameter of 30-50 nm were prepared via an in situ polymerization of aniline in an aqueous solution containing the Fe(3)O(4) magnetic fluid. The inductive heat property of Fe(3)O(4)/PANI composite nanoparticles in an alternating current (ac) magnetic field was investigated. The potential of Fe(3)O(4)/PANI nanoparticles was evaluated for localized hyperthermia treatment of cancers. The saturation magnetization, M(s), and coercivity, H(c), are 50.05 emu g(-1) and 137 Oe for Fe(3)O(4) nanoparticles and 26.34 emu g(-1) and 0 Oe for Fe(3)O(4)/PANI composite nanoparticles, respectively. Exposed in the ac magnetic field for 29 min, the temperatures of physiological saline suspensions containing Fe(3)O(4) nanoparticles or Fe(3)O(4)/PANI composite nanoparticles are 63.6 degrees C and 52.4 degrees C, respectively. The Fe(3)O(4)/PANI composite nanoparticles would be useful as good thermoseeds for localized hyperthermia treatment of cancers.

[A method of showing thermal effect of iron oxide nanoparticles in alternating magnetic field].

BACKGROUND & OBJECTIVE: Ferromagnet has thermal effect in alternating magnetic field (AMF). Magnetically mediating hyperthermia is to localize magnetic substance within tumor tissue under AMF to promote the targeting ability and heat distribution of hyperthermia. This study was to develop a new method of showing the thermal effect of iron oxide Fe3O4 nanoparticles in AMF in vitro. METHODS: Melted polyethelene glycol (PEG) was dropped on a slide to form crystal monolayer after cooling, and was covered with a bipartite blood coverslip. Certain amount of Fe3O4 powder (average diameter, 10 nm) was added in the gap of the blood coverslip to form a strip wandering on the slide with the width of 0.5-1.0 mm. This apparatus, named analog vessel bed, was then alternated in a 4.6 mT AMF at 100-250 kHz for 15 min. The changes of PEG crystal images were observed under micropolariscope. The thermal range of the heated Fe3O4 powder was affirmed according to melting status and melting point value of PEG crystal. RESULTS: In AMF, 5 mg of Fe3O4 could rise the temperature to above 45 degrees C-50 degrees C, which exceeded the required hyperthermic temperature 43 degree C; 15 mg of Fe3O4 could rise the temperature to no more than 59 degrees C-61 degrees C; while 25 mg of Fe3O4 could be heated up to 59 degrees C-61 degrees C. The PEG melting area was enlarged with the increasing Fe3O4 quantity or decreasing PEG melting point. CONCLUSION: The analog vessel bed apparatus could display the thermal effect of Fe3O4 nanoparticles in AMF, and would be helpful for further studies on effect of magnetically mediated hyperthermia on cancer cells.