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We observed nishikinezumi, cinnamon-coloured (NC)/Fujita (F) mice aged between 5 and 28 weeks. These NC mice have skin eruptions that resemble human atopic dermatitis (AD) under conventional circumstances. We investigated the skin of eruptive and non-eruptive lesions in NC/F mice by using haematoxylin–eosin (H&E) staining, toluidine blue staining and immunohistopathological study with immunoglobulin (Ig)EℇRI, CD23, interleukin (IL)-4, IL-5, interferon (INF)-γ and Ia antigen. Histological examination of the eruptive lesions revealed the perivascular infiltration of many lymphocytes and mast cells into the upper dermis. Intracellular oedema of the epidermis, lymphocyte infiltration into the epidermis and liquefaction degeneration of the basal layer were also observed. The numbers of IL-4 and IL-5 positive cells in the eruptive lesions were larger than those of the non-eruptive lesions. IL-4 and IL-5 positive cells in the eruptive lesions increased weekly. Some IFN-γ positive cells were observed in the eruptive lesions after 21 weeks. IFN-γ positive cells were scarce in the skin of both the non-eruptive and eruptive lesions before 21 weeks. Serum IgE increased from 7 weeks to 21 weeks. We confirmed that these findings indicated that T helper (Th)2-dominant immunological activation transformed to a Th1-dominant situation. Many IgEℇRI positive cells were recognized in the dermis of the eruptive lesions by the time IgE had decreased. We assumed that the dermatitis before 21 weeks was an IgE-mediated allergy. We have previously reported that older NC/F mice had positive patch-test reactions to mites. Because serum IgE decreased after 21 weeks, dermatitis after 21 weeks might be associated more with cell-mediated delayed hypersensitivity than with IgE-mediated immediate allergy. [ABSTRACT FROM AUTHOR]

We examined the modifying effects of freeze-dried whole-leaf Aloe arborescens Miller var. natalensis Berger (Japanese name, Kidachi aloe; designated as ‘ALOE’) on 1,2-dimethylhydrazine (DMH)-induced colorectal tumorigenesis in mice. Female ICR mice (7-weeks old) were given a basal diet or a diet containing 1, 0.5 or 0.1% ALOE for 32 weeks. One week later, all mice were injected i.p. with DMH (20 mg/kg, once weekly for 10 weeks) or vehicle (1 mM EDTA solution, pH 6.5). At 32 weeks, animals were killed by exsanguination, and the colorectums were processed for histological examination. The administration of ALOE (1, 0.5 or 0.1% in diet) did not induce diarrhea or reduction of body weight. In mice given DMH and 1% ALOE (Group 2), the incidence and multiplicity of colorectal adenomatous hyperplasias were significantly decreased as compared with mice given DMH alone (Group 1) (both p < 0.05), whereas the incidence and multiplicity of tumors (adenoma and adenocarcinoma) in Group 2 tended to be lower than those in Group 1. In addition, the incidence and multiplicity of the colorectal proliferative lesions (the total of adenomatous hyperplasias, adenomas and adenocarcinomas in mouse colorectum) in Group 2 were significantly decreased as compared with Group 1 (both p < 0.01). No colorectal proliferative lesions were found in animals that did not receive DMH. These results indicated that ALOE reduces the incidence and multiplicity of DMH-induced colorectal proliferative lesions, especially adenomatous hyperplasia, in mice. [ABSTRACT FROM AUTHOR]

An acute leukemia with an unusual immunophenotype developed in a 17-year-old girl. At the initial presentation, extramedullary involvement was not evident, but with advancing disease, massive splenomegaly and an osteolytic rib tumor developed. The disease was aggressive and refractory to intensive chemotherapeutic regimens for myeloid and lymphoid malignancies, and the patient died 3 months after the initial presentation. The leukemic cells were of irregular shape and variable size; they had deeply indented or bi-lobed nuclei and relatively fine, azurophilic granules in their cytoplasm. They were positive for acid phosphatase and beta-glucuronidase in granular staining, but they were negative for myeloperoxidase. The leukemic cells had a unique immunophenotype: it was positive for T-cell antigens (CD1a, CD2, cytoplasmic CD3, CD4), myeloid antigens (CD13 and CD33), NK-cell antigen (CD56), CD19 and CD30. DNA analysis revealed no gene rearrangement in the T-cell receptor beta, gamma and delta, or immunoglobulin heavy chain genes. The leukemic cells of our patient are thought to have arisen from the transformation of a putative precursor cell common to both the T- and NK-cell lineage in the bone marrow. The current literature on precursor NK-cell malignancy is reviewed, and its clinicopathological feature is discussed. [ABSTRACT FROM AUTHOR]

In cases of adult T cell leukaemia neoplastic T cell infiltration in the skin was accompanied by an increase in Langerhans cells. This is in keeping with the view that Langerhans cells may induce antigen-specific and allogenic T cell activation. [ABSTRACT FROM PUBLISHER]

Background: A small percentage of patients with foreign body ingestion develop complications, which have a variety of clinical presentations. Less than 1% of cases require surgical intervention. We present a patient with an abdominal wall abscess resulting from a fish bone that pierced the cecum. The patient was treated laparoscopically., Case Presentation: A 55-year-old Japanese man presented to our hospital with a complaint of right lower abdominal pain. A physical examination revealed tenderness, swelling, and redness at the right iliac fossa. Computed tomography showed a low-density area with rim enhancement in his right internal oblique muscle and a hyperdense 20 mm-long pointed object in the wall of the adjacent cecum. Based on the findings we suspected an abdominal wall abscess resulting from a migrating ingested fish bone. He was administered antibiotics as conservative treatment, and the abscess was not seen on subsequent computed tomography. Two months after the initial treatment, he presented with the same symptoms, and a computed tomography scan showed the foreign body in the same location as before with the same low-density area. We diagnosed the low-density area as recurrence of the abdominal wall abscess. He underwent laparoscopic surgery to remove the foreign body. His appendix, and part of his cecum and the parietal peritoneum that included the foreign body, were resected. He had an uneventful postoperative course, and at 1 year after the surgery, the abdominal wall abscess had not recurred., Conclusions: An abdominal wall abscess developed in association with the migration of an ingested fish bone. We suggest that a laparoscopic surgical resection of the portion of the bowel that includes the foreign body is a useful option for selected cases.

Hepatic carcinosarcoma (HCS) generally presents in advanced stages, demonstrates aggressive behavior, and has a poor prognosis. Other than curative primary resection, no effective treatment options exist. We present a case of resected HCS with four repeat resections for solitary lymph node recurrence followed by chemoradiotherapy with doxorubicin and ifosfamide. A 67-year-old Japanese man was admitted to our hospital for evaluation of an asymptomatic hepatic tumor. The patient underwent right hepatectomy with a presumptive preoperative diagnosis of atypical hepatocellular carcinoma. Based on histopathological and immunohistochemical findings, the tumor was diagnosed as HCS containing osteosarcoma and chondrosarcoma components. After the initial surgery, the patient underwent four additional resections for solitary lymph node HCS recurrence, and then underwent chemoradiotherapy with doxorubicin and ifosfamide for an unresectable lymph node recurrence. Chemotherapy was stopped after two cycles because of severe adverse events, although chemoradiotherapy markedly reduced the size of the lymph node recurrence and provided a progression-free survival of 12 months. Thirty-seven months after the initial surgery, the patient died of cardiac invasion of multiple mediastinal lymph node metastases. The clinical course outlined in this case report suggests that chemoradiotherapy with doxorubicin and ifosfamide for metastatic HCS may prolong survival in patients with unresectable lesions.

Mesenchymal stem cells (MSCs) are pluripotent cells that can differentiate into multilineage cell types, including adipocytes and osteoblasts. Mechanical stimulus is one of the crucial factors in regulating MSC differentiation. However, it remains unknown how mechanical stimulus affects the balance between adipogenesis and osteogenesis. Low intensity pulsed ultrasound (LIPUS) therapy is a clinical application of mechanical stimulus and facilitates bone fracture healing. Here, we applied LIPUS to adipogenic progenitor cell and MSC lines to analyze how multilineage cell differentiation was affected. We found that LIPUS suppressed adipogenic differentiation of both cell types, represented by impaired lipid droplet appearance and decreased gene expression of peroxisome proliferator-activated receptor γ2 (Pparg2) and fatty acid-binding protein 4 (Fabp4). LIPUS also down-regulated the phosphorylation level of peroxisome proliferator-activated receptor γ2 protein, inhibiting its transcriptional activity. In contrast, LIPUS promoted osteogenic differentiation of the MSC line, characterized by increased cell calcification as well as inductions of runt-related transcription factor 2 (Runx2) and Osteocalcin mRNAs. LIPUS induced phosphorylation of cancer Osaka thyroid oncogene/tumor progression locus 2 (Cot/Tpl2) kinase, which was essential for the phosphorylation of mitogen-activated kinase kinase 1 (MEK1) and p44/p42 extracellular signal-regulated kinases (ERKs). Notably, effects of LIPUS on both adipogenesis and osteogenesis were prevented by a Cot/Tpl2-specific inhibitor. Furthermore, effects of LIPUS on MSC differentiation as well as Cot/Tpl2 phosphorylation were attenuated by the inhibition of Rho-associated kinase. Taken together, these results indicate that mechanical stimulus with LIPUS suppresses adipogenesis and promotes osteogenesis of MSCs through Rho-associated kinase-Cot/Tpl2-MEK-ERK signaling pathway.

LPS signaling is mediated through MyD88-dependent and -independent pathways, activating NF-?B, MAP kinases and IRF3. Cot/Tpl2 is an essential upstream kinase in LPS-mediated activation of ERKs. Here we explore the roles of MyD88 and Cot/Tpl2 in LPS-induced chemokine expression by studying myd88(-/-) and cot/tpl2(-/-) macrophages. Among the nine LPS-responsive chemokines examined, mRNA induction of ccl5, cxcl10, and cxcl13 is mediated through the MyD88-independent pathway. Notably, Cot/Tpl2-ERK signaling axis exerts negative effects on the expression of these three chemokines. In contrast, LPS-induced gene expression of ccl2, ccl7, cxcl2, cxcl3, ccl8, and cxcl9 is mediated in the MyD88-dependent manner. The Cot/Tpl2-ERK axis promotes the expression of the first four and inhibits the expression of the latter two. Thus, LPS induces expression of multiple chemokines through various signaling pathways in macrophages., (Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Osteoblasts express Toll like receptor (TLR) 4 and produce osteoclast-activating cytokines in response to the stimulation by lipopolysaccharide (LPS). It has recently been reported that LPS exerts an inhibitory effect on osteoblast differentiation into osteocytes. However, the molecular mechanisms of this inhibitory effect remain ambiguous. The downstream signals of TLR4 are mediated by adaptor molecules including myeloid differentiation factor 88 (MyD88), leading to the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases (ERKs), whose activation by LPS requires the upstream serine/threonine kinase, Cot/Tpl2. To determine the signal molecules responsible for the inhibitory effects of LPS on osteoblast differentiation, we examined the in vitro differentiation of the primary osteoblasts from myd88(-/-) and cot/tpl2(-/-) mice. The matrix mineralization by the wild-type and cot/tpl2(-/-) osteoblasts was significantly inhibited by LPS, whereas that of myd88(-/-) was not affected. During differentiation, LPS suppressed the mRNA expression of runt related transcription factor 2 (Runx2), osterix (Sp7), and activating transcription factor 4 (ATF4) in the wild-type, but not in the myd88(-/-) osteoblasts. The inhibitory effect of LPS on the mRNA expression of these transcription factors was absent in the early phase but partially impaired in the late phase of differentiation in the cot/tpl2(-/-) osteoblasts. Thus, the inhibitory effect of LPS on osteoblast differentiation is Myd88-dependent, whereas the degree of its requirement for Cot/Tpl2 varies depending on the differentiation phase., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Although several murine mAbs that have been humanized became useful therapeutic agents against a few malignancies, therapeutic Abs are not yet available for the majority of the human cancers because of our lack of knowledge of which antigens (Ags) can become useful targets. In the present study we established a procedure for comprehensive identification of such Ags through the extensive isolation of human mAbs that may become therapeutic. Using the phage-display Ab library we isolated a large number of human mAbs that bind to the surface of tumor cells. They were individually screened by immunostaining, and clones that preferentially and strongly stained the malignant cells were chosen. The Ags recognized by those clones were isolated by immunoprecipitation and identified by MS. We isolated 2,114 mAbs with unique sequences and identified 21 distinct Ags highly expressed on several carcinomas. Of those 2,114 mAbs 356 bound specifically to one of the 21 Ags. After preparing complete IgG(1) Abs the in vitro assay for Ab-dependent cell-mediated cytotoxicity (ADCC) and the in vivo assay in cancer-bearing athymic mice were performed to examine antitumor activity. The mAbs converted to IgG(1) revealed effective ADCC as well as antitumor activity in vivo. Because half of the 21 Ags showed distinct tumor-specific expression pattern and the mAbs isolated showed various characteristics with strong affinity to the Ag, it is likely that some of the Ags detected will become useful targets for the corresponding carcinoma therapy and that several mAbs will become therapeutic agents.

Approximately 30 cases of carcinoid tumor of the kidney have been reported in the English literature, including three cases found as components of teratomas. Renal composite tumors associated with somatostatinoma have not been described. A 53-year-old female presented with an incidentally found right renal cystic lesion. Computed tomography demonstrated a cystic lesion associated with a solid nodule in the right kidney and postcontrast dynamic MRI revealed enhancement of the solid nodule. The patient underwent radical nephrectomy for the kidney lesion and is now well without recurrence 21 months after the operation. From the histopathological findings we diagnosed the cystic lesion as a composite tumor composed of mucinous cystadenoma and carcinoid tumor. Immunohistochemistry demonstrated the majority of cells of in carcinoid portion to be positive for antisomatostatin staining. The present case is the first documented composite tumor of mucinous cystadenoma and somatostatinoma of the kidney.

Background: Generally, Langerhans cells deliver antigen information from the skin to the draining lymph nodes via lymph vessels., Methods: By immunohistopathology, we investigated the delivery route of Langerhans cells in human skin using CD1a and S-100 protein antibodies., Results: We noted CD1a- and S-100-positive Langerhans cells in the lymph vessels of the dermis. These were shaped like dendritic cells and presented with some lymphocytes, melanophages, melanin granules and lymph in the same vessels., Conclusion: These observations support the concept that Langerhans cells deliver antigen peptides to regional lymph nodes via afferent lymph vessels., (Copyright 2003 S. Karger AG, Basel)

Objective: To examine 10 cases with primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL), analyze their clinical manifestations and pathological and immunohistochemical features, and improve early diagnosis of this disease., Methods: We studied the morphological characteristics of primary cutaneous CD30-positive ALCL using histopathological methods. Leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelial membrane antigen (EMA), cytokeratin (CK) and HMB45 antibodies were used to determine the expression of their respective antigens from routine paraffin samples of the patients., Results: Ten patients (7 men and 3 women, aged 31 to 84 years) complained of subcutaneous masses or papular eruptions over their lower trunks and extremities. Histopathologically, the lesions were composed of numerous large round or oval pleomorphic cells. The cytoplasm was usually abundant, amphophilic or basophilic, and finely vacuolated. Nuclei were commonly eccentrically localized and lobated or horseshoed in shape, and multinucleated giant cells and Reed-Sternberg-like cells were seen. Nucleoli were generally multiple and large. Of the 10 patients, tumor cells displayed positive antigen expression of CD30 in all cases, positive CD45RO in 6 cases, positive CD20 in only 1 case, but negative CD45RO and CD20 expressions in 3 cases. Two patients died at 7 weeks and 3.4 years of follow-up, respectively., Conclusion: Our study highlights the importance of histopathologic features and positive CD30 staining for differentiation of this disease from other malignant skin tumors.

We report a rare case of contact urticaria due to polyethylene gloves. The patient, a 46-year-old cook, had had had chronic urticaria since 1985, and first visited our hospital in June 2000. We began by prescribing antihistamine and antiallergenic drugs for him, but his condition did not improve. From a detailed interview, we established that when he put on polyethylene gloves at work, his condition worsened. We suspected some component of his gloves to be the cause of his symptoms. Prick and scratch tests with a solution extracted from his gloves showed a wheal-and-flare reaction at 15 min. We advised him to wear a cotton shirt under his clothes in daily life, and to put on cotton gloves under his polyethylene gloves while at work. Subsequently, the size and the number of wheals were markedly smaller and the subject's symptoms were reduced.

Background: Ketoprofen, suprofen and tiaprofenic acid are arylpropionic anti-inflammatories. Their chemical structures share the same elements as the benzoyl radical and the tiophene ring. We experienced nine cases of ketoprofen photoallergy, seven cases of suprofen photoallergy and three cases of tiaprofenic photoallergy., Purpose: To find the key structure of photosensitivity and photocross-reactivity to ketoprofen, suprofen and tiaprofenic acid., Methods: : Three animals were tested for phototoxicity and six animals for the photosensitization potentials of ketoprofen, suprofen, tiaprofenic acid and benzophenone, and the photocross-reactivity of the above chemicals. Test substances were applied symmetrically on both sides of the animals' backs. The animals were irradiated with 180 mJ/cm2 UVB ((1/2) MED) and 10 J/cm2 UVA on the left side. The reactions were read on days 2, 3 and 4. The photosensitization potentials of ketoprofen, suprofen, tiaprofenic acid and benzophenone were determined using the Adjuvant-Strip method. Six animals were assigned to each test group and to a control group., Results: Ketoprofen, suprofen, tiaprofenic acid and propionic acid showed negative reactions with the phototoxic test. Benzophenone showed phototoxic reactions to 40% acetone (ac.), 20% ac. and 10% ac. Therefore, we used 5% aq. benzophenone with the photosensitization test. Ketoprofen was the strongest photosensitizer (6/6) and showed photocross-reactivities to suprofen (2/6), tiaprofenic acid (3/6) and benzophenone (6/6). Suprofen was a strong photosensitizer (4/6) and showed photocross-reactivities to ketoprofen (1/4) and tiaprofenic acid (2/4), but not to benzophenone. Tiaprofenic acid was also a photosensitizer (2/6) but showed a photocross-reactivity only to benzophenone (2/2). Benzophenone was also the strongest photosensitizer (6/6), but did not photocross-react to the above three chemicals., Conclusion: From the test results, it appears that benzoyl radical is the key structure for photosensitivity and the photocross-reactivity of ketoprofen, suprofen and tiaprofenic acid. The whole structure of benzophenone was needed to induce photosensitization of benzophenone. The animals that were photosensitized from the entire structure of benzophenone did not photocross-react to ketoprofen, suprofen or tiaprofenic acid.

We previously reported a case of contact urticaria syndrome (CUS) due to di(2-ethylhexyl) phthalate (DOP) in a polyvinyl chloride (PVC) grip on cotton gloves. The patient reported in this previous paper was careful not to have any contact with PVC products in his daily life or in his working environment. He discontinued the use of protective gloves with a PVC grip that was the cause of CUS. When working, he used cotton gloves without a PVC grip. We prescribed antihistamines which slightly improved his condition. However, when he wore work clothes while on duty, CUS relapsed. This condition was severe and made him feel anxious. When we advised him to wear a cotton shirt under his work clothes, the contact urticaria did not develop. We suspected that some component of the work clothes was the cause of his symptoms. A prick test with the extract solution of his work clothes showed a wheal and flare at the 15 min reading. The common component of the grip and the work clothes was found by analysis to be DOP.

The restriction endonuclease SmaI has been used for the diagnosis of neurogenic muscle weakness, ataxia and retinitis pigmentosa disease or Leigh's disease, caused by the Mt8993T-->G mutation which results in a Leu156Arg replacement that blocks proton translocation activity of subunit a of F(0)F(1)-ATPase. Our ultimate goal is to apply SmaI to gene therapy for this disease, because the mutant mitochondrial DNA (mtDNA) coexists with the wild-type mtDNA (heteroplasmy), and because only the mutant mtDNA, but not the wild-type mtDNA, is selectively restricted by the enzyme. For this purpose, we transiently expressed the SmaI gene fused to a mitochondrial targeting sequence in cybrids carrying the mutant mtDNA. Here, we demonstrate that mitochondria targeted by the SmaI enzyme showed specific elimination of the mutant mtDNA. This elimination was followed with repopulation by the wild-type mtDNA, resulting in restoration of both the normal intracellular ATP level and normal mitochondrial membrane potential. Furthermore, in vivo electroporation of the plasmids expressing mitochondrion-targeted EcoRI induced a decrease in cytochrome c oxidase activity in hamster skeletal muscles while causing no degenerative changes in nuclei. Delivery of restriction enzymes into mitochondria is a novel strategy for gene therapy of a special form of mitochondrial diseases., (Copyright 2002 National Science Council, ROC and S. Karger AG, Basel)

We examined the modifying effects of heated garlic (Allium sativum L.) on N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG)-induced duodenal and jejunal carcinogenesis in mice. Heated garlic powder used in this study was prepared as follows: unpeeled garlic bulbs were blanched in boiling water for 6 min, and then peeled, the cloves being crushed, homogenized, and finally freeze-dried. The garlic powder had almost undetectable alliinase activity and was rich in alliin (the main sulfur compound of heated garlic; 22.1 &mgr;/g dry weight). Male C57BL/6 mice were given ENNG (100 &mgr;/l) in drinking water for the first 4 weeks, and then basal diet (Group 1), or 10% (Group 2), 3% (Group 3) or 1% (Group 4) heated garlic in the diet for 30 weeks. At the termination of the experiment, the incidences of duodenal tumors in Groups 1-3 were significantly lower than those in Group 1, and the multiplicities in Group 2 were significantly lower than those in Group 1. Additionally, the incidences and/or multiplicities of the jejunal tumors in Groups 2 and 4 were also significantly lower than those in Group 1. In this study, we also examined changes in erythrocyte polyamine levels. Values for Group 1 were significantly greater than those in the control group, and this elevation in Group 1 were significantly inhibited by dietary heated garlic (10% in the diet; Group 2). These results indicated that the post-initiation-stage feeding of heated garlic, especially at 10% in the diet, inhibits ENNG-induced duodenal and jejunal carcinogenesis in mice.

An 84-year-old man presented with complaints of epigastric discomfort. Upper gastrointestinal series and endoscopy showed an elevated lesion at the posterior wall of greater curvature on the gastric fundus. Diagnosed as moderately differentiated tubular adenocarcinoma by biopsy, wedge resection and 4sa regional lymph node dissection were carried out. The tumor morphology showed type I with slight elevation, 2.5 x 1.7 cm in size; histological showed papillary, tubular, and solid formations having clear cytoplasm and large bizarre nuclei invading the deep submucosal layer (sm2). This case was evaluated as T1(sm) N0 M0 stage Ia early gastric cancer. In the 5th month after operation, multiple liver metastases were detected. He died of liver failure by rapid growth of metastatic tumors in the 6th month after operation. The serum alpha-fetoprotein level at recurrence was 1,900 ng/mL, and alpha-fetoprotein-positive cells were immunohistochemically detected in operative and liver biopsy specimens.

We examined the PLD activities of human renal cancers and found that the PLD2 activity was greatly elevated in almost all cases examined as compared with the adjacent normal region. Western blot analysis showed the increased levels of PLD2 protein, but the PLD1 was not discernible. The oleate-dependent PU) activity was very low but appeared to increase in most cases. Interestingly, the immunohistochemical observations indicated the high expression of PLD2 in the nuclei of clear carcinoma cells. This is the first demonstration which suggests the possible involvement of PLD2 in tumorigenesis of renal cancer.

We examined the modifying effects of freeze-dried whole-leaf Aloe arborescens Miller var. natalensis Berger (designated as 'ALOE') on N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG)-induced duodenal tumorigenesis in C57BL/6 mice. Experiment 1: Male mice were given ENNG in drinking water for the first 4 weeks, and then 10% ALOE in basal diet for 16 weeks. Experiment 2: Female mice were given ENNG for 5 weeks, and then 5%, 1% or 0.2% ALOE in the diet were given for 15 weeks. In Experiment 1, the tumor incidence and tumor multiplicity (tumors per mouse) of the duodenum in the ENNG + 10% ALOE group were significantly decreased compared with that in the ENNG alone group. Erythrocyte polyamine levels in the ENNG + 10% ALOE group were also significantly decreased. In Experiment 2, the incidence of duodenal tumors in the ENNG + 5% ALOE group were significantly decreased compared with that in the ENNG alone group. These results indicated that ALOE, especially at 10% in the diet, inhibits ENNG-induced duodenal tumorigenesis in mice.

For gene therapy of human malignant glioma, we adopted positively charged multilamellar liposomes entrapping the human interferon beta (hIFN-beta) gene. One week after the transplantation of human malignant glioma U251-SP cells to produce glioma in nude mouse brain, the liposomes entrapping the gene (500 ng of DNA per 25 nmol of lipids per 2 microl) were injected into the same site of the cell transplantation once every second day for a total of five injections; and by this means the tumor completely disappeared. To confirm the antiproliferative effect of hIFN-beta, we performed an in vitro study using a plasmid containing a secretion signal sequence-deleted hIFN-beta gene and one containing the hIFN-beta gene inserted in reverse. In both cases, there was no hIFN-beta release into the medium and no growth inhibition effect. On addition of anti-hIFN-beta antibody to the medium, the growth inhibition effect was abolished. As this cell line expresses IFN-alpha/beta receptor, the hIFN-beta produced in the transfected cells could be released and acted in a paracrine manner. For 120 days the body weight change of normal mice treated by the same procedure as used in the curing experiment was not significant among the groups injected with empty liposomes, plasmid only, and liposomes entrapping the gene. In all of these three groups, death, abnormal behavior, and significant histological changes were not observed.