原 著
組織カラードップラー法による前駆出期および拡張早期における左室局所心筋運動の評価とその有用性

中島　淑江
埼玉医科大学第二内科学教室 〔平成14年11月8日 受付〕

Utility and Assessment of Regional LV Function during Pre-ejection Contraction Period and Early Filling Period by Tissue Color Doppler Imaging Method
Yoshie Nakajima (Second Department of Internal Medicine, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　To evaluate regional LV function, regional pre-ejection contraction time (PECT), and pre-ejection contraction velocity (PECV) and early diastolic velocity (Emyo vel) were measured by M-mode tissue color Doppler in normal myocardium of 28 normal patients (pts), hypertrophic myocardium of 18 pts with hypertrophic cardiomyopathy and infarcted myocardium of 11 pts with myocardial infarction. Data were compared with global LV isovolumic contraction time (ICT) and early diastolic mitral flow velocity (Emv) measured by pulsed Doppler. PECT was significantly shorter than ICT in all patients. PECT was uniform in LV of the normal group. However,the mean PECT was shorter in hypertrophic myocardium, and longer in infarcted myocardium than that in normal myocardium. In both hypertrophic and infarcted myocardium, abnormalities of PECV and Emyo vel were detected easily while LV-ICT or Emv was within normal. M-mode tissue color Doppler methods are useful in distinguishing between normal, hypertrophic and infarcted myocardium more sensitively.
Keywords:tissue color Doppler, pre-ejection contraction time, asynchrony, regional LV function, regional myocardial velocity
J Saitama Med School 2003;30:1-8
(Received November 8, 2002)

原 著
ラットにおけるセロトニン誘発気管支収縮に対するサルポグレラートの作用

田中　 求
埼玉医科大学薬理学教室〔平成14年11月12日 受付〕

The Effects of Sarpogrelate on Serotonin-Induced Bronchoconstriction in Rats
Motomu Tanaka (Department of Pharmacology, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　A bolus injection of 5-HT (32μg/kg) into the right atrium immediately induced Bezold-Jarisch reflex (hypotension, bradycardia and apnea) and delayed bronchoconstriction (assumed by decreased air flow and increased intrapleural pressure) in anesthetized, spontaneously breathing rats. This bronchoconstrictor response was mimicked by α-methyl-5-HT (5-HT₂-receptor agonist : 32μg/kg), and 5-methoxytryptamine (5-HT₂- and 5-HT₄-receptor agonist : 32μg/kg), but not by 1-phenylbiguanide (5-HT₃-receptor agonist : 16μg/kg) or cisapride (5-HT₄-receptor agonist : 32μg/kg). To study the effects of 5-HT and α-methyl-5-HT on the airway dynamics, lung conductance (G_L) and lung compliance (C_L) were calculated from flow rate, tidal volume and intrapleural pressure curves. Dose response curve (DRC) of 5-HT on G_Lor C_Lwas not affected by granisetron (5-HT₃-receptor antagonist : 0.05 mg/kg), but DRC of α-methyl-5-HT was significantly shifted to the right. Subsequently, both the DRCs of 5-HT and α-methyl-5-HT were shifted to the right by sarpogrelate (5-HT₂-receptor antagonist : 0.3, 1.0 mg/kg) in a dose-dependent manner. Moreover, the bronchoconstriction of 5-HT and α-methyl-5-HT (respectively 32μg/kg) were either abolished or potently reduced by bilateral, cervical vagotomy, but the bronchoconstriction response to 5-HT was partly recovered at 128μg/kg. Peak response latency of G_Lwere 9.14±0.39 sec in 5-HT (32μg/kg), suggesting that the response might be originated at the bronchial vascular bed. These results suggest that, at least in the rat, 1) 5-HT-induced bronchoconstriction is mediated by 5-HT²-receptor, but not by 5-HT₃- or 5-HT₄-receptor, 2) sarpogrelate antagonizes 5-HT-induced bronchoconstriction in a dose-related manner, 3) 5-HT-induced bronchoconstriction is potently augmented by vagal reflex, but is primarily mediated by a direct action in the airway, 4) the possible site of action of intravenous 5-HT is the bronchial, not the pulmonary vascular bed.
Keywords:Sarpogrelate, 5-hydroxytryptamine (5-HT), 5-HT₂receptor, Vagotomy, α-Methyl-5-HT, Bronchoconstriction
J Saitama Med School 2003;30:9-18
(Received November 12, 2002)

原 著
レニン-アンジオテンシン系抑制薬は透析療法を受けている糖尿病性腎症患者の左室肥大を抑制する

金子　敬子
入間台クリニック（〒358-0031 埼玉県入間市新久816）〔平成14年12月9日 受付〕

Left Ventricular Hypertrophy (LVH) and Inhibition of Renin-Angiotensin System with ACE Inhibitor (ACEi) or Angiotensin Receptor Antagonist (AIIA) in Diabetic Patients on Dialysis Therapy
Keiko Kaneko ( Irumadai Clinic, Iruma-shi, Saitama 358-0031, Japan )

Background:LVH is frequently found in diabetic and hypertensive patients at the initiation of dialysis therapy and is highly predictive of future cardiac morbidity and mortality. However, it is unknown whether the blockage of renin-angiotensin system will regress LVH or not in these patients using ACEi or AIIA or both of them in combination.Method:Twenty four diabetic patients (62±3 years old; F/M: 8/16) with end-stage renal disease who had just entered into hemodialysis therapy and were diagnosed as having LVH evaluated by echocardiography were selected from 5 dialysis units staffed by the faculty of Saitama Medical School, Saitama, Japan between 1996 and 1998. The study was carried out for 2 years. An ACEi, enalapril 5 mg daily or an AIIA, losartan 50 mg daily was assigned at random. These drugs were administered 30 min after the cessation of dialysis therapy and usually given after lunch when dialysis therapy was not received. The doses of these drugs were adjusted up to 10 mg or 100 mg daily respectively according to the levels of blood pressure. One year after the start of the study, if LVH was not regressed in spite of treatments with enalapril or losartan, add-on each other drug was done.Results:Using repeated measures analysis of variance, applied to those with four echocardiograms, there were progressive decreases over time in LV mass index (LVMi: g/m2), posterior wall thickness, left ventricle end-diastolic diameter, interventricular thickness. The biggest changes in LVMi and other parameters found between the baseline and year 1 (147±14 vs. 138±12 g/m2). Combination therapy of enalapril and losartan for 7 patients who did not respond the initial therapy produced a significant reduction of LVH at the end of study. There was no significant difference in regression of LVH as well as blood pressure control and other variables between enalapril and losartan. Furthermore, there were significant correlations between the variables obtained from the repeated echocardiography related with LVH and systolic blood pressure in all patients.Conclusion:Antihypertensive treatment with either ACEi or AIIA is beneficial in the regression of LVH in diabetic patients who started dialysis therapy. Moreover, a combination therapy with ACEi and AIIA would provide a favorable effect on LVH in those patients unless a single administration of ACEi or AIIA is effective.
Keywords:Left ventricular hypertrophy, Renin angiotensin system, combination therapy
J Saitama Med School 2003;30:19-28
(Received December 9, 2002)

原 著
IL-4遺伝子プロモーター多型と1型糖尿病との関連

大久保　智子
埼玉医科大学第4内科学教室〔平成14年12月10日 受付〕

A Novel Polymorphism in the Promoter Region of the Interleukin-4 Gene and Association with Type 1 Diabetes Mellitus
Tomoko Ohkubo (Fourth Department of Internal Medicine, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　Interleukin-4 (IL-4), one of the key regulators of the Th1-Th2 balance, promotes the differentiation of CD4＋helper lymphocytes into Th2 cells. It is assumed that the Th1 predominance over Th2 has an important role in the development organ-specific autoimmune diseases, such as type 1 diabetes and Hashimoto's thyroiditis. In the present study, therefore, we investigated the possible role of the IL-4 gene in type 1 diabetes and autoimmune thyroid disease. First, we screened for variations in the promoter region ( position－1,105 to＋38;＋1＝translation start site) and exons of the IL-4 gene in 24 patients by PCR direct sequencing, and detected five polymorphisms: C(－589)T, C(－144)T, C(－33)T, A363T and A8411C. Among them, A363T and A8411C are located in introns and were not further analyzed. Genotyping of other polymorphisms were performed by PCR-RFLP on 114 patients with type 1 diabetes, 92 patients withGraves'disease, 57 patients with Hashimoto's thyroiditis and 221 control subjects. C(－144)T is a novel and relatively rare polymorphism, and it was revealed that the distribution of the C(－144)T genotypes was significantly different between type 1 diabetic patients and control subjects (p＝0.0035); the CT genotype was observed in six type 1 diabetic patients and one control subject, and the remainder were all CC genotype. The frequency of the CT genotype was also increased in patients with autoimmune thyroid disease compared to control subjects, but the difference was not significant. No significant difference was observed in the distribution of C(－589)T and C(－33)T genotypes between patients with type 1 diabetes or autoimmune thyroid disease and control subjects. Then, effects of the C(－144)T polymorphism on promoter activity of IL-4 gene were assessed by luciferase reporter gene assays. Transcriptional activity of the－144T allele, both stimulated and unstimulated, was significantly lower than that of the－144C allele. These data suggest that the C(－144)T polymorphism in the IL-4 gene reduces the IL-4 synthesis and contributes to the development of type 1 diabetes.
Keywords: type1 diabetes, interleukin-4 (IL-4), promoter, polymorphism, association
J Saitama Med School 2003; 30:29-36
( Received December 10, 2002)

原 著
尿細管上皮細胞におけるConnective Tissue Growth Factor（CTGF）発現の検討

小林　竜也
埼玉医科大学腎臓内科学教室〔平成14年12月10日 受付〕

Expression of Connective Tissue Growth Factor in Renal Tubular Epithelial Cells
Tatsuya Kobayashi (Department of Nephrology, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

Background/Purpose:Connective tissue growth factor (CTGF) is a member of CCN family which mediates profibrotic effects of transforming growth factor-β1 (TGF-β1), promoting fibroblast proliferation and extracellular matrix (ECM) production. CTGF has been presumed to involve in a variety of organ fibrogenesis. In the kidney at health and a disease, by using in situ hybridization glomerular parietal and visceral epithelial cells, mesangial cells, and interstitial fibroblasts were shown to express CTGF. Recently, tubular epithelial cells have been also revealed to express CTGF especially in diabetic nephropathy. Therefore, in this study, I evaluated CTGF protein expression in the tubular epithelium of renal biopsy specimens by immunohistochemistry, and the expression regulation and the role of CTGF in the cultured tubular epithelial cells (mPTEC).Methods:Using an anti-CTGF antibody, I performed catalyzed signal amplification immunohistochemistry on renal biopsy specimens from patients with minimal change nephrotic syndrome (MCNS), diffuse proliferative lupus nephritis (DPLN), IgA nephropathy (IgAN), and diabetic nephropathy (DN). Using mPTEC cultured in monolayer and co-culture with renal fibroblasts (TFB), whether a given humoral factor could induce CTGF mRNA expression and whether CTGF derived from mPTEC could stimulate TFB to produce type I collagen were tested. The expression of mRNA and type I collagen were determined by ribonuclease protection assay and indirect enzyme-linked immunosorbent assay, respectively.Results:Significant CTGF expression in the tubular epithelium was found in parallel to the interstitial fibrosis in DN samples. In addition, it was revealed that glucocorticoid (GC) therapy resulted in significant induction of tubular CTGF expression in MCNS and DPLN. Among profibrotic growth factors and proinflammatory cytokines employed, only TGF-β1 could induce CTGF mRNA expression in mPTEC in time- and dose-dependent fashion. D-glucose and dexamethasone could also induce CTGF expression in mPTEC. CTGF derived from mPTEC by TGF-β1 and d-glucose could subsequently induce type I collagen production in TFB.Conclusion:Renal tubular epithelial cells can express CTGF in vivo and in vitro, which is likely involved in renal fibrogenesis at least in DN and other diseases undergoing GC therapy.
Keywords:connective tissue growth factor (CTGF), tubular epithelial cells, transforming growth factor-β(TGF-β), d-glucose, glucocorticoid (GC)
J Saitama Med School 2003;30:37-49
(Received December 10, 2002)

原 著
閉経後高血圧の診断方法と治療法の検討

小林　和裕
埼玉医科大学腎臓内科学教室〔平成14年12月10日 受付〕

Study I:This study was carried out to determine whether non-invasive assessment of cardiovascular system discriminates white coat hypertension (WCH). The major reason is the high prevalence of WCH in these subjects and it remains uncertain whether WCH is associated with cardiovascular risk. Women were required to be naturally menopausal; at least 1 year, but not greater than 5 years, past their menstrual period. Exclusion criteria were past history of preeclampsia or eclampsia, severe illness such as myocardial infarction and stroke within 6 months, having used estrogens or progestins within 3 months, being treated with antihypertensive agents or with non pharmacological therapy such as reduction of daily salt, and proteinuric nephropathy. In addition, secondary hypertension was excluded as possible. WCH was defined if subjects who had BP more than 150/90 mmHg at office had to have both mean systolic and diastolic pressures below 120/80 mmHg at home. Forty-four subjects, mean age 52 years, recruited from the outpatients clinic, were examined. BPs at office and home, pulse wave velocity (PWV), and augmentation index (Aix) were recorded. Left ventricular diameter, septal wall thickness, and left posterior wall thickness were assessed by M-mode echocardiography after selecting the measurement section by B-mode echocardiography. Twenty patients were diagnosed as having WCH based on the criteria in the trial. We found that the patterns of pulse wave were different between subjects with WCH and those with hypertension. The most prominent difference was the levels of Aix. In subjects with WCH, Aixs were 25.6±3.8 compared to 38.6±4.5 mmHg in those with hypertension. Moreover, PWV of subjects with WCH were 6.05±0.53 and those of with hypertension were 8.45±0.72 m/sec (p＜0.01). In addition to these findings, there was a significant association between the values of home systolic BP but not those of office systolic BP and PWV, Aix and LV mass index. When we combined the data of LV mass index and Aix, white coat hypertension was easily segregated from hypertension. The combination of self BP monitoring, echocardiographic data and Aix and PWV would be powerful indicators for treatment of hypertension in postmenopausal women.
Study II:To assess the efficacy of the various classes of antihypertensive drugs in postmenopausal women with hypertension. Women were required to be naturally menopausal; at least 1 year, but not greater than 5 years, past their menstrual period. Exclusion criteria were past history of preeclampsia or eclampsia, severe illness such as myocardial infarction and stroke within 6 months, having used estrogens or progestins within 3 months, and proteinuric nephropathy as well as surgically menopausal. 114 women who participated in this study after informed consent were obtained. These women were diagnosed as having hypertension based on BP of more than 140/90 mmHg at clinic as well as on self-reported BP of more than 130/85 mmHg at home. If both levels of BP were not satisfied with the criteria, such patients were also excluded. All antihypertensive medications were withdrawn 6 weeks before the initiation of the study. All patients were randomly assigned in equal numbers to the following therapeutic groups; A) losartan, 50 mg daily and trichlormethiazide 2 mg twice a week and B) cilnidipine, a calcium channel blocker, 5 mg and arotinolol, an αβ blocker, 10 mg daily. Three groups were retrospectively segregated according to pulse pressure (PP) at the start of the study: Group I (n＝ 24); more than 65 mmHg: Group II (n＝58); below 64 to more than 45 mmHg and Group III (n＝32); below 44 mmHg. In Group I, combination therapy with B) reduced systolic BP much more compared to that with A) (169±2/88±5 to 133±2/73±5 vs 169±2/88±5 to 149±2/66±5 mmHg, p＜0.05). On the other hand, in Group III, A) decreased diastolic BP as well as systolic BP (152±2/106±2 to 123±1/78±1 vs 149±2/107±2 to 129±2/84±1 mmHg, p＜0.05). In Group II, there was no significant difference between two antihypertensive regimens. Other three biological markers were not influenced by any type of treatment. PP before starting medication of hypertension may be useful for the choice of antihypertensive drugs.
Combining these two studies, it is suggested that evaluation and treatment of hypertensive postmenopausal women would be much more investigated than previously and be needed much work to be carried out.
Keywords:White coat hypertension, Pulse wave velocity, Pulse pressure
J Saitama Med School 2003;30:51-60
(Received December 10, 2002)

Original
An Analysis of Death Rates in Tianjin, China, 1991-1996

Feng Yanru
Department of Public Health, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan

Background:In a previous study of the death rates in Tianjin in 1995, the author reported a high death rate from heart diseases and a low death rate from malignant neoplasms, compared with the corresponding rates recorded from other cities in China and Japan. Examination of the geographical variations of the death rates revealed clustering of deaths from cerebrovascular diseases and malignant neoplasms. The objective of the present study was to examine the mortality patterns in detail using data collected over the relatively long period of 1991-1996.Methods:The mortality statistics in Tianjin for the period of 1991-1996 were provided by the Department of Health, Tianjin, China. Population data were obtained from the censuses conducted in 1990 and 1995. The age-adjusted death rates, age-specific death rates and the standardized mortality ratios(SMRs) in the 18 districts of Tianjin were calculated for the three leading causes of death by sex.Results:The total number of deaths in Tianjin during the period 1991-1996 was 303,737. Of these, 63.7 % was attributed to the three leading causes of death. The age-adjusted death rates and age-specific death rates from heart diseases and malignant neoplasms decreased for both men and women during the study period. However, during the same period, the age-adjusted death rate, and for the most of part, the age-specific death rate from cerebrovascular diseases remained at a stable level in both sexes. Analysis of the geographical distribution of the death rates showed that the SMRs for heart diseases were high in the Northwest, the SMRs for cerebrovascular diseases were high in the eastern part of the urban area of Tianjin and in the eastern part of Tianjin, and those for malignant neoplasms were high in the urban and coastal areas. The geographical patterns of deaths from cerebrovascular diseases and malignant neoplasms were consistent with those indicated in the previous study.Conclusion:The present analysis determined the geographical patterns of deaths from the three leading causes of death within Tianjin, and a decreasing trend was apparent in the death rates from heart diseases and malignant neoplasms, while the death rate from cerebrovascular diseases remained stable during the study period. The results of the current study indicated that measures for the prevention and control of non-communicable chronic diseases in Tianjin, particularly of cerebrovascular diseases, were scarcely effective in the 1990s. The results also emphasize the need for special intervention in specific target districts of Tianjin.
Keywords:three leading causes of death, heart diseases, cerebrovascular diseases, malignant neoplasm, age-adjusted death rate, age-specific death rate, standardized mortality ratio (SMR)
J Saitama Med School 2003; 30: 113-120
(Received December 14, 2002)

Original
A Novel Oligosaccharide on Bovine Peripheral Myelin Glycoprotein P0

Jianhong Yan, Kunio Kitamura, and Masahiko Nomura
Department of Physiology, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan

Abstract:Sulfated oligosaccharides have found in glycoproteins in myelin in the PNS, such as P0 and PASII/PMP22. These glycoproteins are involved in the interaction of cell membranes and play important roles in the formation and maintenance of myelin sheaths. These functions are due in part to the oligosaccharides in their molecules. Recently, the structures of the oligosaccharides of bovine peripheral myelin P0 were reported, and some of these carry the HNK-1 carbohydrate epitope (SO₄-3GlcAβ1-3Galβ1-4GlcNAc). In this paper, we identified a novel multisulfated oligosaccharide, MS2, using one-dimensional¹H-NMR, mass spectrometry, and chemical and enzymatic analyses. MS2, which contains a HNK-1 epitope, was from a single N-glycosylation site of P0 glycoprotein. The structure of the novel oligosaccharide was: Manα1-3Manα1-6[(3SO₄)GlcAβ1-3(6SO₄)Galβ1-4(6SO₄)GlcNAcβ1-2Manα1-3]Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc. In addition to the 3-O-sulfate present in the HNK-1 epitope, the oligosaccharide contained a 6-O-sulfated N-acetylglucosamine residue and a 6-O-sulfated galactose residue. This implies that both Gal6-O-sulfotransferase, which has not been reported in the PNS, and GlcNAc 6-O-sulfotransferase are active in the PNS tissues.
Keywords:Myelin, Glycoprotein, P0, Sulfated oligosaccharide, HNK-1, sulfotransferase.
J Saitama Med School 2003; 30: 121-129
( Received January 15, 2003)

原 著
新しい方法を用いた，モルモットの咳に対する中枢性鎮咳薬の効果の検討

河本　定則^1），田中　求^2），坂本　芳雄^1）
1）埼玉医科大学第二内科学教室呼吸器科
2）埼玉医科大学薬理学教室
〔平成15年2月6日 受付〕

Evaluation of Effects of Central Antitussives on Cough Motion by New Method in Guinea Pig
¹⁾Sadanori Kawamoto,²⁾Motomu Tanaka,¹⁾Yoshio Sakamoto (Pulmonary Division, Second Department of Internal Medicine¹⁾, Department of Pharmacology²⁾, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　The present study was conducted to evaluate the effects of dextromethorphan and codeine on the elements of the coughing motion under the condition of upper respiratory inflammation. Coughs were induced by microinjection of citric acid into the upper trachea of conscious and unrestrained guinea pig. Cough numbers were counted and cough expiratory peak flows, expiration time and inspiratory peak flows were measured by a one-chambered whole body plethysmograph. The percent inhibitions of the values by intraperitoneal administration of the antitussives were assessed. Cough numbers were reduced by both drugs. But cough expiratory peak flows were reduced more effectively by administration of dextromethorphan. Cough expiration time and inspiratory peak flow were not affected significantly by either dextromethorphan or codeine. These results suggest that there are differences between dextromethorphan and codeine in terms of effects to cough pattern generator. It would be important to assess the percent inhibition of cough expiratory peak flow as well as those of cough numbers when evaluating the efficiency of an antitussive.
Keywords:Dextromethorphan, Codeine, Citric acid, Microinjection, Cough
J Saitama Med School 2003; 30: 131-137
(Received February 6, 2003)

原 著
フローサイトメトリーを用いた新たな移植後免疫モニタリング法に関する基礎研究

淺野　博，小川　展二，小山　勇
埼玉医科大学消化器・一般外科（I）〔平成15年2月7日 受付〕

New Monitoring Method Using Flow Cytometry for Evaluating Immunosuppression in Transplant Patients with Multiple Combination Therapy
Hiroshi Asano, Nobuji Ogawa, Isamu Koyama (Division of Gastroenterological and General Surgery, Department of Surgery, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan )

　The combination therapy of carcinurin inhibitor (CNI), and inhibitor of nucleic acid production, has been widely utilized in clinical kidney transplantation. Mycophenolate mofetil (MMF) has a strong immunosuppressive effect as compared with azathioprine. As a result, over immunosuppression is often experienced under the combination therapy of MMF and CNI. It is very difficult to adjust each immunosuppressive drug without knowing how much the immunosuppression and how much each drug affects the immunosuppression. The method of monitoring the immunosuppression has not yet become available for transplant patients under the combination therapy. We investigated the usefulness of flow cytometry of the lymphocyte stimulation test under the immunosuppressive drug in vitro. The peripheral blood from four healthy volunteers was separated by density gradient centrifugation. After lymphocytes were dyed with 5(and 6)-carboxy fluorescein diacetate succinimidyl ester(CFSE), cells were cultured with Staphylococcus enterotoxin B (SEB) for 4 days under the Mycophenolic acid (MPA) and/or Cyclosporine (CsA) in various concentrations. After cultured, Division Index (DI) was measured by FACS, and Stimulation Index (SI) was calculated using DI. From a series of multiple concentrations of MPA or CsA, SI 25, 50 and 75 of MPA groups were 54, 18 and 6 ng/ml, and SI 25, 50 and 75 of CsA groups were 250, 100 and 30 ng/ml. We defined the medium concentration of each drug as MPA 30 ng/ml and CsA 100 ng/ml, then 1/5 of these results as low concentration and concentration 5 times as high (setting up MPA: 6, 30 and 150 ng/ml, and CsA: 20, 100 and 500 ng/ml). The mean S.I. of MPA group was 71.27±11.55 at 6 ng/ml, 23.63±13.44 at 30 ng/ml and 4.87±1.33 at 150 ng/ml. The mean S.I. of CsA group were 78.02±9.05 at 20 ng/ml, 44.14±4.89 at 100 ng/ml and 16.51±5.17 at 500 ng/ml. At the combination of low doses of CsA and MPA, S.I. was 61.03±8.03. The addition of a medium dose of MPA to the low dose of CsA decreased S.I. to 12.23±0.75, which is almost the same result of the high dose of CsA group. These results have us conclude that the dose of CsA could be decreased when the medium dose of MMF is combined with CsA treatment. In addition, patterns of the histogram in Flow cytometry found that there were differences between CsA and MPA, leading to a better understanding as to which drug contributed to the over- immunosuppression.
Keywords:Flow cytometry, Monitoring of immunosuppression, Mycophenolic acid, cyclosporine A, CFSE
J Saitama Med School 2003;30:139-145
(Received February 7, 2003)

原 著
HPV感染による子宮頚部病変の長期予後について

王　驤
埼玉医科大学産婦人科学教室〔平成15年2月14日 受付〕

The Prognosis of the HPV Infected Cervical Lesions after Followed-up for a Long Term
O Jou (Department of Gynecology and Obstetrics, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

Objective:HPV infections are very common in women, only very few develop clinically relevant dysplastic lesions or even cancer. Some infected cases experienced spontaneous regression, and some showed persistent infection. On the other hand, cervical dysplasia that displays a certain persistent process from the normal to tumorigenic state in the tissues, is a precancerous lesion on the histopathological viewpoint. Unfortunately, long-term follow-up cases on HPV-infected uterocervixes are limited. In this study, patients with cervical dysplasia were monitored with the attempt of detecting the continuous changes in cervical lesions. Furthermore, treatments for the HPV-infected cases in progression group were attempted and progresses in these cases were observed.Materials and methods:We examined 573 women aged 16-81 who visited our Department of OB/Gyn between April 1992 and October 2002 for Papanicolaou Smear Testing and HPV examination. All the patients were divided into 3 groups. Progression group includes patients with developed process of cervical lesions, which were from smear ≦ Class II to mild dysplasia, or from mild to severe dysplasia, or from dysplasia to carcinoma in situ. Regression group consisted of patients with regression, or reducing from dysplasia to smear ≦ Class II or from severe to mild dysplasia. In the dysplasia cases, patients whose lesions persisted in one stage were classified in persistence group. Results: Of 206 women who were followed-up for 2 years, 118(57.3%) were HPV-positive. Among these, the numbers of the three groups were 56(47.5%) in regression group, 35(29.6%) in persistence group, and 27(22.8%) in progression group. Among 29 women infected with HPV-16, their distribution in the three groups were 5(17.2%), 12(41.4%) and 12(41.4%) respectively. Eighty-eight (42.7%) cases of 206 women were HPV-negative and their distribution in the three groups were 57(64.7%), 26(29.5%), and 5(5.7%), respectively. Of the post-operation cases, 16 were followed-up and their HPV persistencies were reevaluated. Within these 16 cases, 14 were turned out to be HPV-negative and got a smear ≦ Class II.Conclusion:The results confirm an important pathogenic role of high-risk-types HPV infections in cervical dysplasia as well as in cervical cancers. After an effective treatment, HPV-positive cases have a strong tendency of negative conversion, and most of them have an HPV-negative report and normal histologycal findings finally.
Keywords:HPV types, high-risk-type HPV, progression group, cervical dysplasia, carcinoma in situ
J Saitama Med School 2003;30:147-153
(Received February 14, 2003)

原 著
腹膜透析患者の予後検討　－心血行動態に焦点を置いた5年間の経過観察－

正田　純子
埼玉医科大学腎臓内科学教室〔平成15年3月10日 受付〕

The Clinical Prognosis of the Patients Undergoing Continuous Ambulatory Peritoneal Dialysis (CAPD)
-5 Years Retrospective Study Focusing on Cardiovascular Status-
Junko Shoda (Department of Nephrology, Saitama Medical School, Moroyama, Iruma-gun, Saitama, 350-0495 Japan)

　Continuous ambulatory peritoneal dialysis (CAPD) was developed by Popovich and Moncrief in 1975, and gradually spread over the world. There are about 9000 patients receiving CAPD in our country, and it is far small number comparing with other countries or patients receiving hemodialysis. The reasons of that have been discussed as social, customary, and economical characteristics in medicine in Japan. Therefore, almost no institution exists which manages over 100 patients receiving CAPD except our hospital. Twenty-nine patients starting CAPD in 1997 were observed retrospectively for 5 years in our hospital. Eleven of 29 patients withdrew from CAPD for the reason of repeated peritonitis, heart failure, cerebrovascular attack, dialysis failure, and so on. However, there is no predictive factor for withdrawing from CAPD at the start. Peritonitis, diabetes mellitus, and ages over 65 years old that have been thought as a risk of technical or life survival in the patients receiving CAPD, did not effect on the technical or life prognosis in our patients. Our clinical data was not completely compatible with the previous report from other countries. However, the clinical data from 5 years observation of the patients started CAPD in the same period, and managed in one institute with the united clinical protocol, would have a value to the area of peritoneal dialysis in Japan.
Keywords:clinical course, peritoneal dialysis, prognosis
J Saitama Med School 2003; 30:155-162
(Received March 10, 2003)

原 著
降圧薬の腹膜機能に及ぼす影響：人および実験動物での検討

友利　浩司
埼玉医科大学腎臓内科学教室〔平成15年3月10日 受付〕

Effects of Antihypertensive Drugs on Peritoneal Function in Human and Experimental Animals
Koji Tomori (Department of Nephrology, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, JAPAN)

　Antihypertensive therapy for the patients receiving continuous ambulatory peritoneal dialysis (CAPD) has been carried out without any guideline or clear results of large scale study. To investigate the effects of antihypertensive agents on peritoneal function, animal experiment and clinical observation were carried out. ＜Study1＞ Renovascular hypertension in dogs was induced with silver clips on both renal arteries that created 90% occlusion. After confirmation of elevation of blood pressure, usually 20 days after operation, the abdomen was opened while the animals were under general anesthesia. Using a CCD camera, diameters of the small arteries of the peritoneum were measured after oral administration of placebo (n＝5); 8 mg of CS866, a selective angiotensin II type I receptor blocker (n＝5); 10 mg of temocapril, an angiotensin converting enzyme inhibitor (n＝5); or 10 mg of amlodipine, a calcium antagonist (n＝5). Blood pressure was decreased in dogs with CS866 and a similar decrease was observed with the use of other drugs. The diameter of the small vessels was increased in dogs with CS866 and temocapril, respectively, compared with the calcium antagonist.＜Study2＞ The effects of valsartan, on blood pressure, dialysate volume, and residual renal and peritoneal function in 36 CAPD patients before and 3 months after oral administration valsartan 40-80 mg daily were investigated comparing with amlodipine. Blood pressure was decreased significantly. Drain volume in peritoneal dialysis and a weekly peritoneal creatinine clearance was significantly increased. No significant changes in serum creatinine and hemoglobin were observed in both groups. In conclusion, these data clearly demonstrated that the blockade of the renin-angiotensin system produces an increase in solute clearance in hypertensive dogs with mild renal insufficiency. These data indicate that such blockade may be applicable as therapy for hypertensive patients on CAPD.
Keywords:angiotensin type 1 receptor blocker, calcium antagonist, peritoneal function, hypertension
J Saitama Med School 2003;30:163-170
(Received March 10, 2003)

Thesis
細胞間接着によるGab-1のチロシンリン酸化

篠原　将彦
埼玉医科大学総合医療センター第一内科学教室
（指導：吉田　行雄教授）

医学博士　甲第860号　平成15年3月28日 （埼玉医科大学）

　Gab-1はHGF/SF受容体であるc-Metや，EGFレセプター等の受容体型チロシンキナーゼによってチロシンリン酸化を受けるマルチプルドッキングタンパク質である．今回我々は，E-カドヘリンによる細胞間接着機構を解析するため，Gab-1のチロシンリン酸化について検討した．細胞間接着はGab-1のチロシンリン酸化を増加させ，その破綻はGab-1のチロシンリン酸化を減少させた．抗E-カドヘリン抗体は細胞間接着依存性のGab-1のチロシンリン酸化を減少させ，E-カドヘリンの発現はGab-1のチロシンリン酸化を増加させた．選択的なSrcファミリーキナーゼの阻害剤は細胞間接着依存性のGab-1のチロシンリン酸化を減少させ，この酵素の活性を阻害するC-terminal Src キナーゼのドミナントネガティブ変異体の強制発現はGab-1のチロシンリン酸化を増加させた．細胞間接着の破綻はGab-1のチロシンリン酸化を減少させ，同時に細胞間接着に反応するMAPキナーゼやAkt活性を減少させた．これらの結果からE-カドヘリン依存性の細胞間接着は，SrcファミリーキナーゼによってGab-1のチロシンリン酸化を増加させ，Ras/MAPキナーゼとPI3キナーゼ/Aktカスケードの活性を調節していることが示された57)．

Thesis
好酸球の基底膜通過ならびに活性酸素産生に及ぼす好中球の作用

菊地　泉
埼玉医科大学第2内科学教室
（指導：西村　重敬教授）

医学博士　甲第862号　平成15年3月28日 （埼玉医科大学）

Effects of Neutrophils on the Trans-basement Membrane Migration and Superoxide Anion Generation of Eosinophils
Izumi Kikuchi (Pulmonary Division, Second Department of Internal Medicine, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

Eosinophils are generally recognized as central effector cells in airway inflammation of bronchial asthma. On the other hand, neutrophils have been suggested to contribute to acute exacerbations or development of chronic severe disease of asthma. The mechanisms how neutrophils contribute to these pathophysiologic processes in asthma remains to be elucidated, however, neutrophils may affect either accumulation or functional status of eosinophils. The objective of this study was to evaluate whether the presence of neutrophils modifies trans-basement membrane migration and effector function of eosinophils. Eosinophils and neutrophils were isolated from the peripheral blood of healthy subjects. Eosinophil migration across basement membrane was evaluated using Matrigel®-coated chemotaxis chambers in the presence or absence of neutrophils. In the presence of neutrophils, trans-basement membrane migration of eosinophils in response to IL-8 was significantly augmented ( % migration; 12.9±3.1 vs. 1.9±0.5 by control, p＝0.007, N＝10). Similarly, eosinophil migration to GRO-α was also augmented in the presence of neutrophils. The enhanced migration in the presence of neutrophils was partially but significantly inhibited by PAF-antagonists, WEB2086 or WEB2170, and LTB4 receptor antagonist, BIIL260, suggesting the role of PAF or LTB4. Finally, the presence of both neutrophils and eosinophils significantly increased superoxide anion generation in response to IL-8 as compared with that of eosinophils or neutrophils alone.
These results suggest that neutrophils can augment both trans-basement membrane migration and respiratory burst of eosinophils. This pathomechanism may contribute to the eventual manifestation of eosinophilic inflammation in the asthmatic airways.
Keywords:eosinophils, neutrophils, trans basement membrane migration, superoxide anion

Thesis
胃癌関連モノクロナール抗体の肺癌ヘの応用

下野　暢隆
埼玉医科大学第2内科学教室
（指導：西村　重敬教授）

医学博士　甲第845号　平成15年1月24日 （埼玉医科大学）

Application of Human Gastric Cancer-reactive Monoclonal Antibodies to Human Lung Cancer
Nobutaka Shimono (Division of Pulmonology, Second Department of Internal Medicine, Saitama Medical School Moroyama, Iruma-gun, Saitama 350-0495, Japan )

Background:Detection and identification of tumor-associated antigens are essential for immunotherapy as well as diagnosis of tumors. Recently mouse monoclonal antibodies (mAbs) against human gastric cancer were raised in our laboratory. Purpose: We asked if these mAbs detect antigens not only of gastric carcinoma cells but also of lung carcinoma cells, and if there are any difference in the localization of tumor-associated antigens among four major histologically different carcinomas of lung. Method: Mice were immunized with human carcinoma cells and normal cells of stomach. Five mAbs (1e2, 3e3, 5b10, 7c4, 7e4) recognizing cell-surface and intracellular molecules of gastric carcinoma cells but not of normal gastric mucosal cells were established by subtractive immunization. Human lung carcinoma cell lines ( 3 squamous cell carcinomas, 2 adeno carcinomas, 3 small cell carcinomas and 6 large cell carcinomas) and two normal human fetal lung fibroblast cell lines were employed. Cell surface and intracellular expression of antigens recognized by mAbs were examined by flow cytometry. In order to examine the intracellular expression, cell surface was treated with Cytofix/Cytoperm® kits to allow mAbs permeabilize into cells. Results: All lung carcinoma cell lines and or lung fibroblast cell lines were positively stained by all five mAbs. Monoclonal Abs were divided into three categories from staining patterns; 1) 3e3 and 7e4 stained both the surface and the inside of both all carcinoma cells and fibroblast cells, 2) 1e2 and 5b10 stained the inside of all carcinoma cells and the surface of carcinoma cells except large cell carcinomas, and 3) 7c4 stained the inside but not the surface of all carcinoma cells. These results revealed that three out of five mAbs raised against human gastric carcinoma cells can also react specifically with human lung carcinoma cells. The findings that two mAbs (1e2, 5b10) reacted with corresponding antigens on the surface and the inside of squamous cell carcinoma, adenocarcinoma and small cell carcinoma, and reacted with an antigen only inside the cells of large cell carcinoma suggest that those mAbs would be diagnostically useful to discriminate large cell carcinoma from other carcinomas. The analysis of corresponding antigens recognized by these three mAbs is needed to testify its usefulness in both the basic and clinical research in the future.
Keywords:Lung cancer, Monoclonal antibody, Large cell carcinoma

Thesis
舌診の研究

張　立也
埼玉医科大学公衆衛生学教室
（指導：永井　正規教授）

医学博士　乙第839号　平成15年1月24日 （埼玉医科大学）

　中医学と現代西洋医学では人体生理，病理の認識方法や診断，治療の理念が大きく異なっている．舌を観察することによって病気に関する情報を収集する舌診は中医学では一般的に使われているが，西洋医学では舌の診察が狭い範囲でしか重視されていない．本研究は舌質，舌苔，舌態の三つの視点から舌を観察する新たな舌診分類体系を開発し，集団別にそれぞれの舌象の特徴を確認した．一般人に多く見られる舌象は淡紅舌，薄白苔であり，加齢により淡白舌が増加する傾向が見られた．さらに舌象と自覚症状との関連を検討し，舌診による疾病の早期発見・診断・治療に根拠を提供する．今回の集団の観察から，淡白舌，紅舌，紫舌，舌尖部の赤点，大舌，歯痕舌，厚苔など様々な所見に注目することの有用性が示唆された．舌診は人体の健康状態，疾病情報を調べるには直観，迅速，有用な方法と考えられ，日常生活ならび臨床実践の場で重視するべきである．

Inspection of Tongue’s Study
Zhang liYE (Division of Pulmonology, Second Department of Public health, Saitama Medical School Moroyama, Iruma-gun, Saitama 350-0495, Japan )

Keywords:Traditional Chinese Medicine, the inspection of tongue, tongue proper, tongue coating, tongue state, tongue picture

Thesis
甲状腺癌細胞における亜鉛の細胞毒性

皆川　晃伸
埼玉医科大学第4内科学教室
（指導：片山　茂裕教授）

医学博士　甲第857号　平成15年3月28日 （埼玉医科大学）

The Effect of Zinc on Human Thyroid Cancer Cell Line
Akinobu Minagawa (Fourth Department of Internal Medicine, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　Zinc is an essential component of a wide variety of metalloenzymes, transcription factors and other proteins. Intracellular homeostasis of zinc is believed to be critical because of its different biological roles. To attain homeostasis under different conditions, cell must adjust the rate of zinc uptake and efflux, binding to intracellular and extracellular proteins or other molecules, and sequestration into vesicles or organelles. This suggests that proteins involved in controlling such processes would be regulated directly by zinc. The expression of zinc transporters in human thyroid cancer cell line is not unknown.
　Zinc has been reported to have potent cytotoxic effect on thyroid cancer cells. Zinc induced necrotic cell death predominantly rather than apoptotic cell death in thyroid cancer cells. In this study, it was demonstrated that the expression of the antiapoptotic proeteins (Bcl-2, Bcl-xL, phosphorylated Bad, and phosphorylated Akt) were increased, whereas the expression of the proapoptotic proteins (Bad and Bax) were decreased following zinc exposure in 8505C thyroid cancer cells. Phosphoinositide 3-kinase (PI3K) inhibitors inhibited the phosphorylation of Akt induced by zinc, suggesting that zinc activates PI3K followed by phosphorylation of Akt and Bad. Phosphorylation of Akt and Bad may prevent the thyroid cancer cells from apoptotic cell death.
　The expression of zinc transporter (ZnT)-1 and-4 was demonstrated in 8505C thyroid cancer cells by northern blot anarysis. Because ZnT-1 is a membrane protein that transports zinc out of cells, it is of interest that the expression of ZnT-1 mRNA in 8505C was significantly lower than other human cancer cell lines. In contrast, the expression of ZnT-4 mRNA in 8505C thyroid cancer cell line was markedly increased as compared with other human cancer cell lines. The expression of ZnT-2 and ZnT-3 was not observed even by the polymerase chain reaction (PCR) method. Zinquin, a specific fluorescent probe for zinc, showed an increase in intracytoplasmic zinc concentrations after zinc exposure in 8505C.
　In summary, the potent cytotoxic effect of zinc on the thyroid cancer cell line may be due to a decrease in the expression of ZnT-1 that leads to an increase in intracytoplasmic zinc concentration. Although zinc has potent cytotoxic effect, zinc also activates antiapoptotic proteins, especially phosphorylated Akt and Bad, which may prevent the cells from apoptotic cell death.
Keywords:zinc, zinc transportor, thyroid cancer, apoptosis, Akt

Thesis
フローサイトメトリーを用いた新たな移植後免疫モニタリング法に関する基礎研究

淺野　博
埼玉医科大学消化器・一般外科（I）
（指導：小山　勇教授）

医学博士　甲第853号　平成15年3月28日 （埼玉医科大学）

New Monitoring Method Using Flow Cytometry For Evaluating Immunosuppression in Transplant Patients with Multiple Combination Therapy
Hiroshi Asano (Division of Gastroenterological and General Surgery, Department of Surgery, Saitama Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　The combination therapy of carcinurin inhibitor (CNI), and inhibitor of nucleic acid production, has been widely utilized in clinical kidney transplantation. Mycophenolate mofetil(MMF) has a strong immunosuppressive effect as compared with azathioprine. As a result, over immunosuppression is often experienced under the combination therapy of MMF and CNI. It is very difficult to adjust each immunosuppressive drug without knowing how much the immunosuppression and how much each drug affects the immunosuppression. The method of monitoring the immunosuppression has not yet become available for transplant patients under the combination therapy. We investigated the usefulness of flow cytometry of the lymphocyte stimulation test under the immunosuppressive drug in vitro. The peripheral blood from four healthy volunteers was separated by density gradient centrifugation. After lymphocytes were dyed with 5(and6)-carboxy fluorescein diacetate succinimidyl ester(CFSE), cells were cultured with Staphylococcus enterotoxinB(SEB) for 4days under the Mycophenolic acid(MPA) and/or Cyclosporine(CsA) in various concentrations. After cultured, Division Index (DI) was measured by FACS, and Stimulation Index (SI) was calculated using DI. From a series of multiple concentrations of MPA or CsA, SI 25, 50 and 75 of MPA groups were 54, 18 and 6 ng/ml，and SI 25, 50 and 75 of CsA groups were 250, 100 and 30 ng/ml. We defined the medium concentration of each drug as MPA 30 ng/ml and CsA 100 ng/ml, then 1/5 of these results as low concentration and concentration 5 times as high (setting up MPA: 6, 30 and 150 ng/ml, and CsA: 20, 100 and 500 ng/ml). The mean S.I. of MPA group was 71.27±11.55 at 6 ng/ml, 23.63±13.44 at 30 ng/ml and 4.87±1.33 at 150 ng/ml. The mean S.I. of CsA group were 78.02±9.05 at 20 ng/ml, 44.14±4.89 at 100 ng/ml and 16.51±5.17 at 500 ng/ml. At the combination of low doses of CsA and MPA, S.I. was 61.03±8.03. The addition of a medium dose of MPA to the low dose of CsA decreased S.I. to 12.23±0.75, which is almost the same result of the high dose of CsA group. These results have us conclude that the dose of CsA could be decreased when the medium dose of MMF is combined with CsA treatment. In addition, patterns of the histogram in Flow cytometry found that there were differences between CsA and MPA, leading to a better understanding as to which drug contributed to the over- immunosuppression.
Keywords:Flow cytometry, Monitoring of immunosuppression, Mycophenolic acid, cyclosporine A, CFSE

Thesis
マウスおよびラット敗血症性ショックに対するココア投与の有効性に関する検討

小野　一之
埼玉医科大学総合医療センター高度救命救急センター
（指導：堤　晴彦教授）

Thesis
酸化型ガレクチン-1による神経因性疼痛の抑制
－ラット坐骨神経切断モデルを用いた検討－

会田　由紀男
埼玉医科大学総合医療センター麻酔科
（指導：宮尾　秀樹教授）

医学博士　甲第866号　平成15年3月28日 （埼玉医科大学）

　神経因性疼痛は末梢神経あるいは中枢神経の機能異常により生じる機能性疼痛である．その発症メカニズムは複雑で治療に難渋することも多い．今回我々はラットを用いて坐骨神経切断モデルを作製し，その坐骨神経切断部に末梢神経再生促進因子である酸化型ガレクチン-1を0.84μg，単回直接投与した．そして末梢神経傷害後の神経再生と神経因性疼痛の発症抑制の関連および酸化型ガレクチン-1の神経因性疼痛治療としての可能性について検討した．
　坐骨神経を切断して酸化型ガレクチン-1を投与した14日後に，自傷行動を判定した．また後根神経節を含む坐骨神経およびその入力相当レベルの脊髄を摘出して，抗サブスタンスP受容体（NK1R）抗体を用いたウェスタンブロット解析と，抗サブスタンスP抗体，抗NK1R抗体，抗P0（末梢神経ミエリン糖蛋白）抗体を用いた免疫組織化学染色を行った．
　酸化型ガレクチン-1投与により自傷行動スコアは有意な低下を示し，さらに坐骨神経切断端の神経線維の伸長促進，抗P0抗体の発現の増加，後根神経節での小型細胞の維持，サブスタンスPの発現の増加，脊髄でのサブスタンスPの発現の増加とNK1Rの発現の減少が認められた．以上のことから酸化型ガレクチン-1が末梢神経切断後に後根神経節の小型細胞の維持を介して神経因性疼痛の発症を抑制する可能性が示唆された．
Keywords:神経因性疼痛，酸化型ガレクチン-1，サブスタンスP，サブスタンスP受容体（NK1R），坐骨神経切断モデル

Thesis
新規ダントロレン誘導体を用いたCa^2＋放出機構に関する解析

木原　康隆
埼玉医科大学薬理学教室
（指導：丸山　敬教授）

医学博士　甲第850号　平成15年3月28日 （埼玉医科大学）

　リアノジン受容体（RyR）を介する筋小胞体（SR）からのCa^2＋放出は骨格筋の生理的収縮において興奮を収縮につなげる必須の過程である．RyRからCa^2＋放出を起こす機構にはCa^2＋-induced Ca^2＋ release（CICR）がよく知られているが，骨格筋の生理的収縮においてはCICR機構ではなく，RyRがCICRとは異なる開口様式（モード）で機能する生理的Ca2＋放出（PCR）機構が働いていると考えられている．すなわち，細胞膜上の電位変化を横行小管（T管）膜に存在するジヒドロピリジン受容体（DHPR）が検知し，その信号をRyRに何らかの方法で伝えることでCa^2＋放出が起きる．しかしPCRの分子的な機序の詳細はまだ不明のままである．CICRを抑制することなくPCRを特異的に抑制する薬物があればPCRの分子的機構の解明に有用と思われる．悪性高熱の治療薬で知られるダントロレン（Dan）は室温でも高温でもPCRを抑制するが，CICRに対しては高温では抑制を示すけれども室温では抑制しないか，しても極めて弱いと言われる．すなわち，DanはPCRとCICRを少なくとも室温においてはある程度区別している可能性が高い．
　今回，Danよりもっと明白にPCRとCICRを区別する薬物を見つける目的で，数種の新しく合成されたDan誘導体の影響を検討した．これらの薬物について骨格筋無傷線維の単収縮（PCR）抑制作用とサポニン処理スキンド･ファイバーでのCICR抑制作用を調べた．またPCRに類似した特徴を有すると思われるclofibric acid（Clof）による Ca^2＋ 放出に対してのこれらの薬物の影響も検討した．その結果，Dan誘導体の中にはDanよりもPCRとCICRをより明瞭に区別するものがあることがわかった．
　なお，これらの実験の結果，Clofによって活性化されたRyR-Ca^2＋放出チャンネルの開口様式とPCRの開口様式が類似していることを裏付ける結果が得られた．

Thesis
吸入麻酔後の健忘に関する行動学的検討

岡本　由美
埼玉医科大学総合医療センター麻酔科
（指導：宮尾　秀樹教授）

医学博士　乙第886号　平成15年5月23日 （埼玉医科大学）

　本研究では，全身麻酔が，動物の学習・記憶行動にどのような影響を及ぼすかについて，複数の学習・記憶課題を適用して様々な角度から検討した．
　実験1では，麻酔処置に先立って長期間にわたる訓練を行ない，これにより獲得された弁別行動に，笑気・酸素・エンフルランによる全身麻酔がどのような影響を持つかを調べた．その結果，麻酔が弁別行動を阻害するという事実は示されなかった．実験2では弁別遂行の行動上の可塑性を検索する系として多用される弁別逆転課題を用い，過去において一度獲得された弁別行動の再獲得の過程に，全身麻酔処置がどのような影響を持つかを検討した．逆転訓練課題の困難さから，最終的にデータを得られた個体数はきわめて少ないが，麻酔群3個体のうち1個体において，再逆転課題の獲得に遅滞がみられた．実験3では，実験1，2のように長期にわたる訓練を要する課題ではなく，比較的獲得の容易な同時弁別課題を用いて，弁別獲得過程における麻酔の効果を検討した．その結果，高濃度麻酔群で1個体のみ，成績が処置前の獲得基準に到達しないものが見られたが，統計的に有意な麻酔の効果は示されなかった．
　結論として，今回の研究では学習の獲得やその維持に麻酔は顕著な影響を及ぼさないことが示された．このことは麻酔の安全性を改めて示すものではあるが，臨床場面で少数ながら報告される，全身麻酔による逆向性健忘の解明に向けて，そのような稀現象を検討する方法論が求められる．

原 著
Benzbromarone (Urinorm®)の代謝・毒性および薬理作用に関する研究

国嶋　千代子¹⁾，井上　郁夫²⁾，及川　寿浩¹⁾，片山　茂裕²⁾
1）鳥居薬品株式会社医薬情報部
2）埼玉医科大学第四内科学教室
〔平成15年8月8日受付〕

The Metabolism, Toxicity and Pharmacological Studies of Benzbromarone (Urinorm®)
Chiyoko Kunishima^a, Ikuo Inoue^b, Toshihiro Oikawa^a, Shigehiro Katayama^b(^aPharmacovigilance Department of Torii Pharmaceutical Co., Ltd, 3-4-1, Nihonbashihoncho, Chuo-ku, Tokyo 103-8439, Japan and^bFourth Department of Internal Medicine, Medical School, Moroyama, Iruma-gun, Saitama 350-0495, Japan)

　Sporadic reports of mild hepatic dysfunction as an adverse reaction of Benzbromarone have appeared in recent years, and in some cases, fulminant hepatitis has developed. We first investigated metabolites of Benzbromarone, using LC/MS, and identified the metabolites of Benzbromarone in vitro. Results showed that human liver S9 metabolites of Benzbromarone consist of two hydroxy bodies, two monohydroxy bodies and five 1’-ketone bodies; previous reports (except that debromination products were not detected) suggest that Benzarone (possibly related to hepatotoxicity) is not formed in humans. We also identified CYP molecular species related to the metabolism of Benzbromarone, by use of a human P450-expressing microsome system. We found that Benzbromarone is metabolized by CYP2C9*1 and 2C9*2, and the main metabolite is 6-hydroxylbenzbromarone. A study of the inhibitory action of Benzbromarone on human P450 molecular species showed that Benzbromarone has especially potent inhibitory action on CYP2C8/9. Therefore, it was clear that Benzbromarone is metabolized by CYP2C9 and inhibits CYP2C8 and CYP2C9. Deaths caused by fulminant hepatitis in patients given the antidiabetic troglitazone (Noscal), a peroxisome proliferator-activated receptor (PPAR)-γ agonist, have recently been reported. It has also been reported that the mechanism of onset is by the induction of hepatocellular apoptosis by excessive increase of PPAR-γ activity. Because Benzbromarone has long been considered to be a peroxisome proliferating agent, we studied the action of Benzbromarone on PPAR-α and PPAR-γ. The results indicated that Benzbromarone is a ligand of PPAR-α. Our results suggest that the hepatic dysfunction associated with Benzbromarone is not caused by apoptosis.
Keywords:Benzbromarone, PPAR, CYP
J Saitama Med School 2003;30:187-194
(Received August 8, 2003)