Induction of T-cell immunity against leukemia by dendritic cells pulsed with total RNA isolated from leukemia cells

Objectives To assess the feasibility and efficacy of eliciting leukemia-specific T-cell responses in syngeneic mice in vitro and in vivo using dendritic cells (DCs) pulsed with total RNA from leukemia cells.Methods DCs generated from bone marrow culture in vitro in the presence of combined cytokines were pulsed with cellular total RNA isolated from cultured L615 cells by cationic lipid 1,2-dioleoyloxy-3-(trimethylammonium) propane (DOTAP). T-cell responses were evaluated by in vitro proliferation, and cytotoxicity assay. And in vivo immune protection and proghosis of mice with leukemia were studied.Results DCs pulsed with total RNA isolated from cultured L615 cells (DCs/RNA) were remarkably effective in stimulating L615-specific T-cell response in vitro, but did not cross-react with other leukemia cells from syngeneic mice. Vaccination of naive mice with viable DCs/RNA vaccine was able to partly protect from challenge with a lethal dose of live L615 cells, leading to low leukemia incidence and overall survival prolongation. Statistically significant survival was also observed in a low lethal dose of L615-bearing mice that received treatment using viable DCs∕RNA vaccine alone, suggesting that systemic administration of IL-2 could enhance the anti-tumor efficacy of leukemia RNA/DCs vaccine.Conclusions These data support the use of DCs/RNA vaccine as a feasible and effective route to elicit leukemia immunity against unidentified leukemia-associated antigens for treatment of leukemia-bearing animals.     

Insulin induces a novel form of postsynaptic mossy fiber long-term depression in the hippocampus

The mechanisms of induction and the site of expression of long-term depression (LTD) at the hippocampal mossy fiber-CA3 synapses are not clear. Here, we show that a brief bath application of insulin induces a novel form of mossy fiber LTD. This insulin-LTD is (1) induced and expressed postsynaptically, (2) entirely independent of synaptic stimulation during insulin application, (3) involving a rise in postsynaptic [Ca(2+)](i) and L-type voltage-activated Ca(2+) channel activation, (4) mechanistically distinct from low-frequency stimulation-induced LTD, (5) dependent on phosphatidylinositol 3-kinase signaling, and (6) associated with a clathrin-mediated endocytotic removal of surface 3-hydroxy-5-methylisoxazole-4-propionic acid receptors from the postsynaptic neurons. Moreover, insulin-LTD is specific to mossy fibers to CA3 pyramidal cell synapses, and is not present at associational commissural synapses. These findings not only support a postsynaptic locus of mossy fiber LTD, but also provide a further link between the AMPA receptor trafficking and the bidirectional expression of long-term synaptic plasticity.     

套筒制动及扎针对小鼠电针实验中镇痛的贡献

目的 :动物电针镇痛研究大都需要将动物制动和扎针 ,人们难以确定所观察到的镇痛作用中有无制动和扎针带来的应激性镇痛。本实验在观察到 2Hz电针对套筒制动的Swiss Webster小鼠具有镇痛作用的基础上 ,试图分析小鼠电针镇痛实验中套筒制动和扎针对镇痛的贡献。方法 :纸板 /布兜制动的动物作为对照组 (n =8) ,此法对动物的制动作用短暂而轻微。塑料套筒制动的实验组动物根据扎入针灸针有否又分为不扎针 (n =8)和扎针 (n =8)两组。以 48℃热水甩尾潜伏期 (TWL)作为测痛指标。结果 :塑料套筒制动的实验组小鼠 ,无论是不扎针还是扎针 ,其TWL均较纸板 /布兜制动组延长。结论 :电针实验中观察到的镇痛 ,除了来源于电流刺激外 ,制动及扎针引起的痛阈升高也可能是重要因素     

Illuminated lens fixator for bimanual pars plana lensectomy

Pars plana vitrectomy and lensectomy are effective methods for removing posteriorly dislocated lens material. We describe an illuminated nucleus fixator that assists in the removal of posteriorly dislocated lens fragments. We tested the efficacy of the illuminated nucleus fixator compared to a standard endoilluminator in removing lens nuclei from the vitreous cavity of a cadaveric eye model. Use of the illuminated nucleus fixator as compared to a standard endoilluminator significantly reduced the time needed to remove lens nuclei from the vitreous cavity. This instrument could penetrate and fixate lens nuclei and could safely stabilize small and intermediate-sized lens fragments against the activated fragmatome. Bimanual lensectomy using the illuminated nucleus fixator significantly shortens the surgical time needed to remove posteriorly dislocated lens fragments. Shortening the surgical time for lensectomy may minimize such complications as retinal tears, trauma, and phototoxicity.     

p14(ARF) modulates the cytolytic effect of ONYX-015 in mesothelioma cells with wild-type p53

ONYX-015 has been reported to kill selectively tumor cells lacking functional p53. Genetic alterations of INK4a/ARF locus, which is a predominant event in malignant pleural mesothelioma, may result in loss of p14(ARF) and subsequent disruption of p53 pathway in cancer cells. In the present study, ONYX-015 was able to kill three mesothelioma cell lines (H28, H513, and 211H) with wild-type p53 but lacking p14(ARF). In contrast, MS-1 mesothelioma cells, which expressed both p53 and p14(ARF), were resistant to ONYX-015. Introducing p14(ARF) gene into the H28 cell, a mesothelioma cell without p14(ARF) expression, significantly increased the resistance of this cell line to the cytolytic effect of ONYX-015. Our results suggest that human mesotheliomas with wild-type p53 yet lacking p14(ARF) are potential candidates for ONYX-015 therapy.     

Unresectable hepatocellular carcinoma treated with radiotherapy and/or chemoembolization

The purpose of our study was to evaluate the outcome, patterns of failure, and toxicity for patients with unresectable hepatocellular carcinoma (HCC) treated with radiotherapy, transcatheter arterial chemoembolization (TACE), or combined TACE and radiotherapy. Forty-two patients with unresectable HCC were treated with combined radiotherapy and TACE (TACE+RT group, 17 patients), radiotherapy alone (RT group, 9 patients), or with TACE alone (TACE group, 16 patients). Mean dose of radiation was 46.9 +/- 5.8 Gy in a daily fraction of 1.8 to 2 Gy, directed only to the cancer-involved areas of the liver. TACE was performed with a combination of Lipiodol, doxorubicin, cisplatin, and mitomycin C, followed by Gelfoam or Ivalon embolization. Tumor size was smaller in the TACE group (mean: 5.4 cm) compared with the TACE+RT group (8.6 cm) and the RT group (13.1 cm) (P = 0.0003). The median follow-up was 24 months in the TACE+RT group, 28 months in the RT group, and 23 months in the TACE group. Survival was significantly worse for patients treated with radiotherapy alone due to the selection bias of patients with more advanced disease and compromised condition in this group. In contrast, the TACE+RT and TACE groups had comparable survival (two-year rates: TACE+RT 58%, TACE 56%, P = 0.69). The local control rate for the treated tumors was similar in the TACE+RT and TACE groups (P = 0.11). The intrahepatic recurrence outside the treated tumors was common and similar between these two groups (P = 0.48). The extrahepatic progression-free survival was significantly shorter for patients in the TACE+RT group than in the TACE group (two-year rates: TACE+RT 36%, TACE 100%, P = 0.002). Seven patients died from complications of treatment. Local radiotherapy may be added to treat patients with unresectable HCC, and the control of progression of the treated tumors was promising even in patients with large hepatic tumors. Survival of patients with combined TACE and radiotherapy was similar to that with TACE as the only treatment, while a significant portion of the patients treated with radiotherapy developed extrahepatic metastasis.     

Genetic polymorphisms of glutathione S-transferases M1 and T1 associated with susceptibility to aflatoxin-related hepatocarcinogenesis among chronic hepatitis B carriers: a nested case-control study in Taiwan

This study was conducted to investigate the modifying effect of glutathione S-transferase (GST) M1 and T1 polymorphisms on aflatoxin-induced hepatocarcinogenesis among chronic hepatitis B virus surface antigen (HBsAg) carriers. A total of 79 HBsAg-positive cases of hepatocellular carcinoma (HCC) diagnosed between 1991 and 1997 were identified and individually matched to one or two HBsAg-positive controls on age, gender, residence and date of recruitment from the same cancer screening cohort in Taiwan. Blood samples were tested for hepatitis B and C viral markers by enzyme immunoassay and for aflatoxin B(1) (AFB(1))-albumin adducts by competitive enzyme-linked immunosorbent assay. GSTM1 and GSTT1 genotypes were determined by PCR. There was a statistically significant relationship between detectable levels of AFB(1)-albumin adducts in serum and risk of HCC among chronic HBsAg carriers, with an adjusted odds ratio (OR) of 2.0 [95% confidence interval (CI) 1.1-3.7]. In addition, the effect of aflatoxin exposure on HCC risk was more pronounced among chronic HBsAg carriers with the GSTT1 null genotype (OR 3.7, 95% CI 1.5-9.3) than those who were non-null (OR 0.9, 95% CI 0.3-2.4). The interaction between serum AFB(1)-albumin adduct level and GSTT1 genotype was statistically significant (P = 0.03). For GSTM1 the effect of aflatoxin exposure on HCC risk in those with the null genotype was also greater (adjusted OR 2.8, 95% CI 1.0-7.8) than in those with the gene present (adjusted OR 1.8, 95% CI 0.8-4.5), but the difference was not significant (P = 0.91). Notably, when the interaction between aflatoxin exposure and GSTT1 genotype was considered, aflatoxin exposure by itself was not a significant determinant of HCC risk among chronic HBsAg carriers. These results demonstrate the importance of gene-environment interactions in the multifactorial development of HCC.     

The expression of G-protein-gated inwardly rectifying K+ channels GIRK1 and GIRK2 mRNAs in the supraoptic nucleus of the rat and possible role involved

The expression of G-protein-gated inwardly rectifying K channels subunits GIRK1 and GIRK2 mRNAs in the supraoptic nucleus (SON) was investigated in the rat by in situ hybridization with non-radioactive dig-labeled cRNA probes. Double-labeled methods were used to study the co-localization of GIRK1 and 2 and oxytocin (OT) and vasopressin (AVP) in the SON. The present study revealed wide and intense expression of GIRK1 and GIRK2 mRNAs with high overlapping in the SON, indicating the heterologous channel of GIRK1/GIRK2 was a major functional channel in the SON. Given that 100% of OT-positive and 95% of (AVP)-positive neurons in the SON expressed GIRK1/GIRK2 mRNAs, it is possible that GIRK1/GIRK2 channel, activated through G-protein coupled receptors, may be involved in the inhibitory regulation of the release of OT and AVP from the SON.