Plasminogen activator inhibitor-1 (PAI-1) gene transfection inhibits the liver metastasis of pancreatic cancer by preventing angiogenesis

Plasminogen activator inhibitor-1 (PAI-1) is a unique type of serine protease inhibitor and one of the key regulators of tumor invasion and metastasis. The purpose of this study was to elucidate the effect of PAI-1 gene transfection on liver metastasis and its mechanism by using the human high liver metastasis pancreatic cancer cell line, SW1990. PAI-1-transfected SW1990 (SW/PAI-1) produced a significantly higher level of PAI-1 in supernatant than parental cells. While no difference was observed for the production of u-PA and u-PA activity in the supernatant, cell proliferation of SW/PAI-1 was slightly suppressed on the 7th day of incubation compared to parental cells. Cellular invasion, in vivo tumorigenesis in xenograft and liver metastasis were significantly suppressed in SW/PAI-1 cells compared to parental cells. The angiogenesis of xenograft by detecting microvascular density and the production of metastasis-related factors, such as VEGF and TGF-beta1, were also decreased in SW/PAI-1 cells. These findings suggested that PAI-1 gene transfection might have the ability to prevent the liver metastasis of pancreatic cancer by modulating angiogenesis.     

Radiation-induced cell death is independent of the apoptotic signals mediated by death-associated protein kinase in human cervical squamous cell carcinoma cells

Death-associated protein kinase (DAPK) was originally identified as a positive mediator of interferon-gamma (IFNgamma)-induced apoptosis in cervical cancer cells, and interferons have been reported to enhance radiosensitivity in various types of squamous cell carcinoma. To examine whether DAPK can regulate cancer cell radiosensitivity, we investigated DAPK expression and radiosensitivity in human cancer cell lines, including the cervical squamous cell carcinoma cell line, ME180, which is both radiosensitive and IFNgamma-sensitive. Of the 5 human cancer cell lines examined, ME180 cells were the most radiosensitive, but their level of DAPK protein expression was undetectable by western blotting. A comparative study of ME180 cells with 2 other uterine cancer cell lines, HHUA and HOKUG, revealed no significant relationships between cellular radiosensitivity and DAPK protein expression or hypermethylation of the DAPK promoter CpG island. INFgamma dose-dependently inhibited ME180 cell proliferation, but did not induce any cell death. IFNgamma significantly enhanced the radiosensitivity of ME180 cells with a slight increase in DAPK protein expression, while irradiation significantly reduced their sensitivity to the growth-inhibitory signals of INFgamma. Analyses of 6 monoclonal cisplatin-resistant subclones established from ME180 cells revealed that all 6 were significantly more radioresistant than the parent ME180 cells without any change in the DAPK protein expression. These results indicate that DAPK does not regulate radiation-induced cell death and that it cannot be either a target molecule for radiotherapy with gene therapy or a prognostic marker for cervical cancer patients treated with radiotherapy.     

Cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a paracrine tissue-specific growth factor(s)

To examine the possibility of a cervical squamous cell carcinoma-specific growth factor(s), previously suggested by clinical findings and in vitro culture experiments, we examined in vitro cultures and nude mouse transplantations of cervical squamous cell carcinoma cells obtained from a patient with a bulky cervical tumor without detectable distant metastases. The cancer cells did not proliferate without additional growth factors but remained viable in vitro. Fourteen weeks after transplantation of the tumor cells on their backs, 1 of the 3 nude mice developed large metastatic pelvic tumors without macroscopic metastatic lesions in their lungs or liver. When these pelvic tumor fragments were trans-planted onto the backs of other nude mice, ulcerated back tumors and larger metastatic pelvic tumors, but no macroscopic metastatic lesions in the lungs or liver, were observed. Histopathological examination of these pelvic tumors showed that all were the same squamous cell carcinoma as the primary tumor. These results indicate that cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a squamous cell carcinoma-specific growth factor(s) in a paracrine manner.     

Efficacy of intraperitoneal continuous hyperthermic chemotherapy as consolidation therapy in patients with advanced epithelial ovarian cancer: a long-term follow-up

This trial was performed to determine the efficacy and progression-free and overall survivals of patients with advanced ovarian cancer who had been treated with intraperitoneal hyperthermic chemotherapy (IPHC). Ten patients with advanced ovarian cancer participated in this trial and were treated with IPHC. The median progression-free and overall survival rates for all patients treated in this study were 41.2 and 70.2 months, respectively. Two of ten patients received optimal primary cytoreduction surgery followed by IPHC; four of ten, optimal interval debulking surgery followed by IPHC; and four of ten, negative second-look operation followed by IPHC. The groups had 5 and 14.5, 17.75 and 38, and 82.75 and 130.25 months median progression-free and overall survival rates, respectively. Grades 3-4 toxicity included myelosuppresion, and nephropathy was detected. One patient required blood transfusions due to grade 4 anemia and thrombocytopenia. Another patient developed grade 3 nephrotoxicity but did not require continuous hemodialysis. IPHC was feasible, produced manageable toxicity, and showed promise for the treatment of advanced ovarian cancer. Negative second-look laparotomy followed by IPHC was especially effective when consolidation intraperitoneal chemotherapy had been indicated. It produced excellent median progression-free and overall survival rates.     

Radiation reduces pirarubicin sensitivity in human cervical squamous cell carcinoma cells

To find an effective protocol for chemoradiotherapy with pirarubicin hydrochloride (THP) for advanced cervical cancer patients, effects of irradiation on THP sensitivity were examined using the radiosensitive human cervical squamous cell carcinoma cell line ME180. Concurrent irradiation significantly reduced THP sensitivity, and this was further reduced when cells were treated with THP 8 h after irradiation. However, the THP sensitivity of cells treated with THP 8 h before irradiation was significantly enhanced. Four months after the first irradiation, 4 subclones of ME180 cells that survived repetitive irradiation demonstrated significantly higher THP sensitivity than non-irradiated parent cells. These results indicate that THP injections, more than several hours before irradiation or after completion of radiotherapy, might be better therapies than concurrent chemoradiotherapy with THP for cervical cancer.     

Survey of hip fractures in Japan: Recent trends in prevalence and treatment

Background

A nationwide survey of hip fractures by the Japanese Orthopaedic Association (JOA) from 1998 to 2008 found a drastic increase in incidence. The aims of this study were to elucidate the status of hip fractures from 2009 to 2014 and to survey the causes for delayed surgery.

Upregulation of IL-13 concentration in vivo by the IL13 variant associated with bronchial asthma

BACKGROUND: A substantial body of evidence exists to support the pivotal role of IL-13 in the pathogenesis of bronchial asthma. We recently found that a variant of the IL13 gene (Arg110Gln) is genetically associated with bronchial asthma, which is concordant with animal experiments using IL-13 in the development of asthma. OBJECTIVE: To address whether the Gln110 variant of IL13 influences IL-13 function, contributing to the pathogenesis of bronchial asthma, we studied the functional properties of the variant. METHODS: We generated 2 types of recombinant IL-13 proteins, the amino acids of which at 110 were arginine or glutamine, and analyzed the binding affinities with the IL-13 receptors, as well as the stability of the proteins. We further compared the relationship between the genotype and serum levels of IL-13. RESULTS: The variant showed a lower affinity with the IL-13 receptor alpha2 chain, a decoy receptor, causing less clearance. The variant also demonstrated an enhanced stability in both human and mouse plasma. We further identified that asthmatic patients homozygous for the Gln110 variant have higher serum levels of IL-13 than those without the variant. CONCLUSION: These results suggested that the variant might act as a functional genetic factor of bronchial asthma with a unique mechanism to upregulate local and systemic IL-13 concentration in vivo.     

Cytotoxic difference of T cells expanded with anti-CD3 monoclonal antibody in the presence and absence of anti-CD 28 monoclonal antibody

Bulk T cells can be expanded by CD3 stimulation alone (CD3-Ts) or by CD3/CD28 dual stimulation (CD3/CD28-Ts) of peripheral blood mononuclear cells (PBMC). However, few reports have described the difference of features between CD3-Ts and CD3/CD28-Ts. PBMC were stimulated with anti-CD3 monoclonal antibody (mAb) alone or co-stimulated with anti-CD3/CD28 mAbs immobilized on plastic plates, in the presence of rhIL-2 for 4 days, subsequently cultured in the presence of rhIL-2 with no antibody then analyzed. The expansion rate was significantly lower for CD3-Ts (965 + 510-fold, n=5) than CD3/CD28-Ts (2263 + 856-fold, n=5) (p<0.05). The CD4/CD8 ratio, the percentage of CD28(+) cell, and the percentage of T cells with no ability to generate intracytoplasmic interleukin-4 (IL-4) or interferon-gamma (IFN-gamma) were all significantly higher, but, phenotypically, memory cells were lower in CD3/CD28-Ts than in CD3-Ts. The levels of activity of both natural killer (NK) and lymphocyte-activated killer (LAK) cells were lower in CD3/CD28-Ts than CD3-Ts. In comparison to CD3-Ts, CD3/CD28-Ts showed impaired migration toward RANTES. In conclusion, T cells expanded with anti-CD3 and anti-CD28 mAbs differ from those expanded with anti-CD3 alone with proliferation, cytotoxicity, chemotaxis, and phenotype. These differences may exert profound influences on the therapeutic potential of output cells.     

Does delayed no-reflow phenomenon cause myocardial necrosis?

Although recent studies have suggested that the no-reflow region progressively extends during the later reperfusion period, whether this “delayed” no-reflow causes myocardial necrosis remains unknown. To examine this question, we analyzed alteration of the size and histology of the no-reflow region and regional contractlie function after ischemia/reperfusion in anesthesized rabbit preparation. The no-reflow zone after 30-minute ischemia was 13.3 ± 4.2% of the ischemic region at 10 minutes after reperfusion and 35.1 ± 6.4% at 60 minutes, which indicates a significant extension of the no-reflow region during a 60-minute reperfusion period. There were both almost-normal-appearing myovytes and necrotic cells with contraction bands in the no-reflow zone at 10 minutes after reperfusion. In contrast, the larger region of no-reflow region after a 60-minute reperfusion consisted mostly of contraction band necrosis, suggesting irreversible ischemic cell injury in this area before reflow. The regional systolic thickening fraction (TF), which was reduced to −86 ± 23% after 30-minute ischemia, recovered to −30 ± 20% of the baseline value after 10 minutes of reperfusion, and there was no deterioration in the TF during the subsequent reperfusion. Furthermore, the response of TF to dobutamine (5 and 10 μ g/kg/min, i.v.), which was infused to eliminate myocardial stunning, did not decrease during the 60-minute reperfusion period. These results suggest that delayed no-reflow does not extend myocardial infarction.     

Ca2+-dependent inward current induced by nicotinic receptor activation depends on Ca2+/calmodulin-CaMKII pathway in dopamine neurons

It is well known that midbrain dopamine (DA) neurons receive massive projection from cholinergic neurons in the brainstem. In our preceding report, we showed that Ca(2+)-influx through nicotinic acetylcholine (ACh) receptors in the DA neurons subsequently activated an inward current that was sensitive to fulfenamic acid (FFA) and phenytoin, presumably a Ca(2+)-activated non-selective cation current. The FFA-sensitive current exhibited a negative slope conductance and predominantly enhanced the depolarizing responses of DA neurons. In this study, we showed that the inward FFA-sensitive current was eliminated by antagonists of Ca(2+)/calmodulin (Ca(2+)/CaM), N-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide hydrochloride (W-7; 1 microM), trifluoperazine (TFP; 1.5 microM) and calmidazolium (100 nM). Application of W-7 and TFP reduced the ACh-induced inward current and the current component suppressed by these drugs exhibited negative slope conductance, as well as the FFA-sensitive current. Further, intracellular application of KN-93, an antagonist of Ca(2+)/CaM-dependent protein kinase II (CaMKII), but not KN-92 eliminated the FFA-sensitive current. All these results suggest that Ca(2+)/CaM-CaMKII pathway is involved in an activation of the FFA-sensitive current.