Hepatectomy and intraarterial infusion chemotherapy for liver metastasis from colorectal cancer

Hepatectomy and intraarterial chemotherapy for liver metastasis from colorectal cancer have been performed in our department. Intraarterial infusion chemotherapy has also been performed for unresectable liver metastasis. One hundred twenty-seven cases of liver metastasis from colorectal cancer were studied. The cases were divided into groups according to radicability of the original colorectal cancer, whether or not hepatectomy was performed, and whether or not they received intraarterial chemotherapy. Group I is cur C of origin. Group II is cur A or B without hepatectomy. Group III is cur A or B with hepatectomy. Each group was divided into a group without intraarterial chemotherapy (A) and a group with it (B). IA 23 cases, IB 13 cases, IIA 14 cases, IIB 21 cases, IIIA 28 cases, and IIIB 28 cases. The survival rate of group III was better than that of group II. The survival rate of group II was better than that of group I. There was no significant difference in survival rates between IA and IB. The survival rate of group IIB was significantly better than that of group IIA. The survival rate of group IIIB was significantly better than that of group III A. Hepatectomy and intraarterial chemotherapy after hepatectomy for liver metastasis from colorectal cancer were effective.     

Prognostic significance of p53, Ki-67, VEGF and Glut-1 in resected stage I adenocarcinoma of the lung

OBJECTIVES: The purpose of this study was to evaluate the prognostic significance of various biological factors in patients with resected stage I adenocarcinoma. METHODS: We immunohistochemically examined 47 specimens of surgically resected adenocarcinomas to evaluate the expression of the biological markers p53, Ki-67, vascular endothelial growth factor (VEGF) and glucose transporter-1 (Glut-1). Angiogenesis grade and tumor vessel invasion was also investigated. Actuarial survival was analyzed by the Kaplan-Meier method. Clinical variables and biological markers were analyzed using the Cox's proportional hazards model for multivariate analysis to identify independent prognostic factors. RESULTS: The overall survival rate for the whole series was 85.1% at 3 years and 71.9% at 5 years, with a median survival time of 73 months. Differentiation, Ki-67, Glut-1, VEGF, tumor vessel invasion and microvessel density (MVD) were significant prognostic factors by univariate analysis, with Glut-1 expression the most important prognostic factor for survival (P<0.0001). After multivariate analysis, only Glut-1 expression remained as a prognostic factor for survival. CONCLUSION: Glut-1 expression can be a predictor for prognosis in patients with resected stage I adenocarcinoma of the lung.     

Oral administration of the thrombin receptor antagonist E5555 (atopaxar) attenuates intimal thickening following balloon injury in rats

Thrombin is a powerful agonist for a variety of cellular responses including platelet aggregation and vascular smooth muscle cell (SMC) proliferation. These actions are mediated by a thrombin receptor known as protease-activated receptor-1 (PAR-1). Recently we discovered that 1-(3-tert-butyl-4-methoxy-5-morpholinophenyl)-2-(5,6-diethoxy-7-fluoro-1-imino-1,3-dihydro-2H-isoindol-2-yl)ethanone hydrobromide (E5555, atopaxar) is a potent and selective thrombin receptor antagonist. This study characterized the pharmacological effects of E5555 on SMC proliferation in vitro and in a rat model of intimal thickening after balloon injury in vivo. E5555 selectively inhibited rat aortic SMC proliferation induced by thrombin and thrombin receptor-activating peptide (TRAP) with half maximal inhibitory concentration (IC(50)) values of 0.16 and 0.038 μM, respectively. E5555 did not inhibit rat SMC proliferation induced by basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) at concentrations up to 1μM. In addition, E5555 inhibited human aortic SMC proliferation induced by thrombin at concentrations of 0.3 and 3units/ml with IC(50) values of 0.028 and 0.079 μM, respectively, whereas it did not affect bFGF-induced proliferation at concentrations up to 1μM. Repeated oral administration of 30 mg/kg E5555 (once daily for 16 days) significantly reduced neointimal formation in the balloon-injured rat arterial model. These results suggested that a PAR-1 antagonist could be effective for treating restenosis following vascular intervention in addition to preventing thrombus formation. E5555 could thus have therapeutic potential for restenosis and chronic atherothrombotic disease.     

Toxicokinetics of bisphenol A in rats, monkeys and chimpanzees by the LC-MS/MS method

We examined the toxicokinetics of bisphenol A (BPA) in F344 rats, cynomolgus monkeys and chimpanzees. Serum BPA levels were quantified using the LC-MS/MS method. After oral administration at 10 mg/kg, the maximum concentration in the serum (C(max)) and the area under the serum concentration curve (AUC) of BPA in cynomolgus monkeys and chimpanzees were greater than in rats. After oral administration at 100 mg/kg, AUC during the first 4h (AUC(0-->4h)) in cynomolgus monkeys was greater than in rats. In rats, the serum BPA levels were increased again 6h or later after oral administration at each dose, which suggested the enterohepatic circulation of BPA in rats. After subcutaneous administration at 10 mg/kg, the AUCs were ranked in the following order: cynomolgus monkeys>chimpanzees>rats, and C(max) in cynomolgus monkeys was greater than in rats and chimpanzees. After subcutaneous administration at 100 mg/kg to cynomolgus monkeys and rats, both the C(max) and AUCs in cynomolgus monkeys were greater than in rats. In all species, the oral administration of BPA resulted in much lower C(max) and AUCs than subcutaneous administration at the corresponding doses, indicating the low bioavailability of oral administration. This result suggests that BPA undergoes an extensive first-pass metabolism in these animal species. AUCs of subcutaneous administration and the AUC (0-->4h) of oral administration in the two primates were greater than that in rats. Because the systemic clearance for BPA is assumed to be dependent on the hepatic blood flow-rate, the high AUCs in primates are considered to be due to the lower systemic clearance by a lower hepatic blood flow-rate in primates than in rats. In addition, the toxicokinetics of the metabolites of BPA were examined. After the oral administration of 10 mg/kg BPA, both C(max) and AUCs of BPA metabolites were ranked in the following order: cynomolgus monkeys>chimpanzees>rats, and the terminal elimination half-life (T(1/2)) in rats was greater than that in cynomolgus monkeys and chimpanzees, suggesting the enterohepatic circulation of BPA in rats. From these results, the systemic clearance of BPA in primates is considered to be close to that in humans due to the similarity of the hepatic blood flow-rate. Furthermore, the major elimination route of BPA metabolites in primates is assumed to be renal excretion, as in humans, because the enterohepatic circulation that was observed in rats was not observed. In conclusion, primates are thought to be served as a valuable surrogate model for the toxicokinetics of BPA in humans.     

Transanal rectal dissection: A procedure to assist achievement of laparoscopic total mesorectal excision for bulky tumor in the narrow pelvis

Background

Laparoscopic approaches for colorectal surgery have been improved recently; however, it is often difficult to achieve total mesorectal excision (TME) for lower rectal cancer laparoscopically because of a narrow pelvis and a thickened mesentery.

Methods

TME was successfully performed in 6 patients (4 men, 2 women) with dissection of the rectum transanally from the anal side of the tumor. The preoperative stage was T3N1M0 in 1 patient and T3N0M0 in 5 patients. The mean body mass index was 29.8 kg/m² (range, 28.7-31.2 kg/m²), and the mean tumor size was 46.5 mm (range, 30-60 mm).

Results

The mean duration of the anal portion of the operation was 64 minutes (56 minutes in women, 79 minutes in men). No complications occurred during surgery or postoperatively.

Conclusion

This technique is a simple and effective procedure for successfully performing laparoscopic TME of lower rectal cancer in patients with bulky tumors, narrow pelvises, and thickened mesenteries.     

Synthetic peptides coupled to the surface of liposomes effectively induce SARS coronavirus-specific cytotoxic T lymphocytes and viral clearance in HLA-A*0201 transgenic mice

We investigated whether the surface-linked liposomal peptide was applicable to a vaccine based on cytotoxic T lymphocytes (CTLs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). We first identified four HLA-A*0201-restricted CTL epitopes derived from SARS-CoV using HLA-A*0201 transgenic mice and recombinant adenovirus expressing predicted epitopes. These peptides were coupled to the surface of liposomes, and inoculated into mice. Two of the liposomal peptides were effective for peptide-specific CTL induction, and one of them was efficient for the clearance of vaccinia virus expressing epitopes of SARS-CoV, suggesting that the surface-linked liposomal peptide might offer an effective CTL-based vaccine against SARS.     

Epidermal barrier dysfunction and cutaneous sensitization in atopic diseases

Classic atopic dermatitis is complicated by asthma, allergic rhinitis, and food allergies, cumulatively referred to as atopic diseases. Recent discoveries of mutations in the filaggrin gene as predisposing factors for atopic diseases have refocused investigators' attention on epidermal barrier dysfunction as a causative mechanism. The skin's barrier function has three elements: the stratum corneum (air-liquid barrier), tight junctions (liquid-liquid barrier), and the Langerhans cell network (immunological barrier). Clarification of the molecular events underpinning epidermal barrier function and dysfunction should lead to a better understanding of the pathophysiological mechanisms of atopic diseases.     

Flaky Tail Mouse Denotes Human Atopic Dermatitis in the Steady State and by Topical Application with Dermatophagoides pteronyssinus Extract

The barrier abnormality, a loss-of-function mutation in the gene encoding filaggrin (FLG), which is linked to the incidence of atopic dermatitis (AD), is a recently discovered but important factor in the pathogenesis of AD. Flaky tail (Flg^ft) mice, essentially deficient in filaggrin, have been used to investigate the role of filaggrin on AD. However, the relevancy of Flg^ft mice to human AD needs to be determined further. In this study, we observed the clinical manifestations of Flg^ft mice in the steady state and their cutaneous immune responses against external stimuli, favoring human AD. Under specific pathogen-free conditions, the majority of Flg^ft mice developed clinical and histological eczematous skin lesions similar to human AD with outside-to-inside skin barrier dysfunction evaluated by newly devised methods. In addition, cutaneous hapten-induced contact hypersensitivity as a model of acquired immune response and a mite extract-induced dermatitis model physiologically relevant to a human AD were enhanced in Flg^ft mice. These results suggest that the Flg^ft mouse genotype has potential as an animal model of AD corresponding with filaggrin mutation in human AD.Atopic dermatitis (AD), which affects at least 15% of children in developed countries, is characterized by eczematous skin lesions, dry skin, and pruritus.^1,2,3 Although the precise pathogenic mechanism of AD is as yet unknown, several accumulated lines of evidence suggest that a defective skin barrier to environmental stimuli may contribute to its pathogenesis. It has long been thought that the barrier abnormality in AD is not merely an epiphenomenon but rather is the “driver” of disease activity.⁴ The evidence for a primary structural abnormality of the stratum corneum in AD is derived from a recently discovered link between the incidence of AD and loss-of-function mutations in the gene encoding filaggrin (FLG). Individuals carrying the FLG null allele variants tend to develop AD.^5,6,7Filaggrin protein is localized in the granular layers of the epidermis. Profilaggrin, a 400-kDa polyprotein, is the main component of keratohyalin granules.^8,9,10 In the differentiation of keratinocytes, profilaggrin is dephosphorylated and cleaved into 10 to 12 essentially identical 27-kDa filaggrin molecules, which aggregate in the keratin cytoskeleton system to form a dense protein-lipid matrix.¹⁰ This structure is thought to prevent epidermal water loss and impede the entry of external stimuli, such as allergens, toxic chemicals, and infectious organisms. Therefore, filaggrin is a key protein in the terminal differentiation of the epidermis and in skin barrier function.¹¹Because AD is a common disease for which satisfactory therapies have not yet been established, understanding the mechanism of AD through animal models is an essential issue.^1,12 Flaky tail (Flg^ft) mice, first introduced in 1958, are spontaneously mutated mice with abnormally small ears, tail constriction, and a flaky appearance of the tail skin, which is most evident between 5 and 14 days of age.¹³ Mice of the Flg^ft genotype express an abnormal profilaggrin polypeptide that does not form normal keratohyalin F granules and is not proteolytically processed to filaggrin. Therefore, filaggrin is absent from the cornified layers in the epidermis of the Flg^ft mouse.^14,15,16Recently, it has been revealed that the gene responsible for the characteristic phenotype of Flg^ft mice is a nonsense mutation of 1-bp deletion analogous to a common human FLG mutation.¹⁵ These mice developed eczematous skin lesions after age 28 weeks under specific pathogen-free (SPF) conditions¹⁷ and enhanced penetration of tracer perfusion determined by ultrastructural visualization,¹⁶ and were predisposed to develop an allergen-specific immune response after epicutaneous sensitization with the foreign allergen ovalbumin (OVA).^15,17 On the other hand, general immunity through intraperitoneal sensitization with OVA was comparable between Flg^ft mice and control mice.^15,17Despite these recent advances, there still remain several issues with Flg^ft mice to be addressed. For example, serial close observation of clinical manifestations in reference to human AD will be informative. It is of value to evaluate the responses to external stimuli relevant to human AD, such as mite extracts, instead of OVA that has been used previously. A comparative study on the skin-mediated contact hypersensitivity (CHS) response and non-skin-mediated delayed-type hypersensitivity response is important to evaluate the impact of barrier dysfunction on immune responses in vivo. In addition, although it has now been determined that the barrier dysfunction is a key element in the establishment of AD, there is no established method to evaluate the outside-to-inside barrier function quantitatively.In this study, we found that Flg^ft mice showed spontaneous dermatitis with skin lesions mimicking human AD in a steady state under SPF conditions: serial occurrence of manifestations as scaling, erythema, pruritus, and erosion followed by edema in this order. We also successfully evaluated outside-to-inside barrier dysfunction in Flg^ft mice quantitatively using a newly developed method. In addition, we determined that the Th1/Tc1-mediated immune response was enhanced by immunization through skin but not through non-skin immunization. Last, we induced severe AD-like skin lesions in Flg^ft mice by application of mites as a physiologically relevant antigen for human AD, which will be an applicable animal model of AD.     

Increased Apoptosis Associated with Depressed Type of Early Intestinal Gastric Cancer

Early gastric cancer can be macroscopically classified into elevated and depressed types. To clarify the relationship between macroscopic appearance of early gastric cancer and apoptosis or cell proliferation, formalin-fixed paraffin-embedded tissue specimens of 44 intestinal-type early gastric cancers were investigated by the TUNEL method and immunohistochemical techniques. Diffuse type was excluded in this study. When tissue sections of gastric cancer were vertically classified into the 3 compartments of laminar, intermediate and basal, the apoptosis index (%) was significantly higher in the basal compartment of depressed type (1.76±2.04, mean±SD) than in the basal compartment of elevated type (0.63±0.81, P=0.01). In depressed type, the apoptosis index (%) was significantly higher in the basal compartment than in the luminar compartment (0.76 ±0.85, P=0.03). Apoptosis-inducing protein, Bax, was expressed more in each of the compartments of depressed type than in those of elevated type, while there were no significant differences in expression of anti-apoptotic protein, Bcl-2, between the two types. Moreover, the apoptosis index (%) of Bax-positive gastric cancer was significantly higher in the basal compartment (P=0.03), compared to that of Bax-negative gastric cancer, while there were no significant differences in apoptosis index (%) in any compartment between Bcl-2-positive and Bcl-2-negative gastric cancers. There were no significant differences in Ki-67 expression, either between the two types, or among the compartments of depressed type. These results indicate that increased apoptosis with excessive expression of Bax in the basal compartment is involved in the morphogenesis of the depressed type in intestinal-type early gastric cancer.     

Effects of Alcohol Consumption on Histological Changes in Chronic Hepatitis C: A Clinicopathological Study

Background: To assess the effects of alcohol on the histological changes in chronic hepatitis type C, we performed histopathological examination on liver biopsy specimens by using a semiquantitative method. Methods: Subjects were 91 patients with chronic hepatitis type C and 32 with alcoholic liver disease. The patients with chronic hepatitis type C were classified into three groups according to the total amount of alcohol intake: nondrinkers, moderate drinkers, and heavy drinkers. For each patient, we evaluated pathological changes of several items and awarded scores from 0 to 2 points, with severe to moderate scoring 2 points, mild 1, and negative 0; the total score was then compared between groups. The evaluated histological changes included virus-related histological changes (VI, inflammatory cell infiltration; V2, lymphoid follicle formation; and V3, bile duct damage) and alcohol-related changes (A1, perivenular fibrosis; A2, stellate/pericellular fibrosis; and A3, fatty change). Results: The total scores of the hepatitis C virus-related histological changes were significantly lower in patients with alcoholic liver disease (ALD group) (p < 0.05). However, we found no significant difference between the different alcohol intake groups. The total score for alcohol-related histological changes significantly increased in line with increases in total alcoholic intake regardless of the presence or absence of hepatitis type C virus infection (p < 0.05). Conclusions: The results suggest that both alcoholic-related liver damage and virus-related liver damage have specific features; in a addition, alcohol was found to have little effect on the histological liver damage observed in chronic hepatitis type C.