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The Kampo (Japanese herbal) medicine "Juzen-Taiho-To" (TJ-48), which was prepared by decocting a concoction (formula), contains ten kinds of herbs and has several immunostimulating activities. In order to determine the contribution of each herbal component, the complement-activating and mitogenic activities of the hot water extract as well as the polysaccharide fraction from each herb were tested. Hot water extracts of Glycyrrhizae radix, Astragali radix, and Atractylodes lanceae rhizoma showed significant mitogenic activity whereas that of Cinnamomi cortex showed potent complement-activating activity. However, the exclusion of any single component herb whether active or not on its own did not result in a loss or an increase of the overall activity of TJ-48. The polysaccharide fraction from Glycyrrhizae radix showed the most potent of both activities among the same fractions from the other nine herbs, and reduced both activities after periodate oxidation, thus indicating that the carbohydrate moiety may contribute to both activities.

The Metabolism of TJN-101 was studied in rats after oral administration of 3H-TJN-101. 1. At 0.75hr after the oral administration of 3H-TJN-101 (4mg/kg) to fasted rats, 30.0%, 18.1%, and 8.5% of the radioactivity was present in the plasma as unchanged TJN101, Met. A III, and Met. A I, respectively. At 1hr after a single dose and at 1hr after the 21st daily oral administration to non-fasted rats, the ratio of unchanged TJN-101 to total radioactivity in the plasma was 18.1% and 19.6%, respectively. 2. In the liver, at 0.75hr after a single oral administration to fasted rats, 15.3%, 26.3 %, and 6.3% of the radioactivity was present as unchanged TJN-101, Met. A-III, and Met. F, respectively. At 1hr after the 21st administration, only Met. A-III was found in the liver, accounting for 16.0% of the total radioactivity. 3. In the bile collected for 24hr after a single oral administration to fasted rats, most of the metabolites were present in the conjugated form, Met. B, Met. F, and Met. D accounted for 41.3%, 9.4%, and 9.2% of the total radioactivity, respectively. 4. Urine, collected for 24hr after a single oral administration, was enzymaticaly hydrolyzed. The percentages of radioactivity corresponding to Met. B, Met. F, and Met. D were 29.6 %, 16.5%, and 9.0%, respectively. The other metabolites accounted for less than 6 % of the radioactivity. After hydrolysis of urine collected for 24hr from non-fasted rats after a single dose or after the 21st daily dose, Met. B was present, accounting for 32.4% and 39.9% of the total radioactivity, respectively. Almost no differences were found in the composition of the other metabolites in non-fasted rats compared with those in fasted rats. 5. Six metabolites were observed in feces collected for 24hr after a single oral administration to fasted rats, and these constituted less than 9 % of the total radioactivity. In feces collected for 24hr after the 21st daily administration to non-fasted rats, 18.0%, 11.2% and 10.9% of total radioactivity was present as Met. B, Met. F, and an unknown metabolite, M1, respectively.

The absorption and excretion of gomisin A (TJN-101) in rats whose livers were injured by carbon tetrachloride (CCl4) were investigated. After intravenous administration of TJN-101 at a dose of 5 mg/kg, the terminal elimination half-life was 1.5 h in the CCl4-treated rats, which was two times that in normal rats. The mean area under the blood concentration-time curve (AUC) value of TJN-101 in the CCl4-treated rats was twice that in normal rats, and this difference was significant (p less than 0.05). Therefore, the total body clearance of TJN-101 in the CCl4-treated rats decreased less than half of that in normal rats. Similar results were observed when it was administered orally. In the CCl4-treated rats, the serum concentration of Met. B, which was identified as a demethylenated substance and one of major metabolites, tended to decrease more than that in normal rats. On the other hand, the cumulative biliary excretion ratio of TJN-101 in 24 h after dosing in the CCl4-treated rats was 2.5 times that in normal rats. The excretion rate of Met. B in the bile in the CCl4-treated rats tended to be delayed. However, the quantitative variance of biliary excretion of Met. B was not found in both groups. The urinary excretion of TJN-101 or Met. B in 72 h after dosing in the CCl4-treated rats was lower than that in normal rats. Similar results were also observed in excretion in the feces.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of cycloheximide, a protein synthesis inhibitor, on hippocampal selective neuronal death was morphologically studied in rats subjected to 10 min forebrain ischemia using a 4-vessel occlusion model. Neuronal damage in the hippocampal CA1 subfield 72 h after ischemic insult was dramatically decreased by the lasting inhibition of protein synthesis through consecutive administration of cycloheximide. Cycloheximide, which was administered once within the first 24 h of recirculation, showed protective action on ischemic cell necrosis and its most potent effect was observed when injected at 12 h of post-ischemia. After 36 h of recirculation, however, treatment with cycloheximide could no longer prevent cell death. The possibility is considered that hippocampal delayed neuronal death following transient ischemia is caused by abnormal protein(s).

"Juzen-Taiho-To" (TJ-48), which is a kampo (Japanese herbal) medicine prepared by decocting a prescription of ten kinds of herbs, has several immunostimulating activities. In order to characterize the active substances for anti-complementary and mitogenic activities, TJ-48 was fractionated. Anti-complementary activity was observed in the water- and methanol-insoluble fraction (F-2) and the crude polysaccharide fraction (F-5), whereas mitogenic activity was only found in F-5. However, other low molecular mass fractions did not show both activities. Methylation analysis indicated that F-2 mainly contained amylopectin-like polysaccharides. Both Pronase digestion and periodate oxidation decreased the anti-complementary activity of F-2, and the beta-amylase-resistant fraction of F-2 still retained the potent anti-complementary activity. When F-5, which has the most potent of both activities, was further fractionated, only the major acidic polysaccharide fraction, F-5-2, showed potent mitogenic activity. Endo-alpha-(1----4)-polygalacturonase digestion showed that F-5-2 mainly contained pectic polysaccharides, and the endo-polygalacturonase treatment of F-5-2 reduced the mitogenic activity but not the anti-complementary activity. F-2 and F-5 each activated the complement system by a different mode of action.

D-phenylalanine (DPA) is known to block the activity of carboxypeptidase, an enzyme which degrades enkephalins, endogenous morphine-like substances. Therefore, it is considered that DPA administered as an inhibiting drug of this degrading enzyme might prolong analgesia induced by acupuncture. 1) Thirty patients suffering from chronic low back pain were treated with acupuncture 30 minutes after the oral administration of 4.0 grams of DPA. The results were: excellent in 7 cases, good in 11, fair in 6 and poor in 6. Cases graded excellent and good were then compared with a placebo group. The effect was increased 26% in the DPA-acupuncture group, which shows no statistically significant difference (P less than 0.1). 2) In 56 patients, tooth extraction was performed under acupuncture anesthesia: 18 had received 4.0 gram of DPA (P.O.) 30 minutes earlier. The results were excellent in 8, good in 6, fair in 3, and poor in 1. The excellent and good cases were compared with 38 placebo group cases. The effect in the DPA-acupuncture anesthesia group was significantly increased by 35% (P less than 0.01). 3) In order to determine the optimum time for the administration of DPA, two schedules of administration were compared. [1] DPA was given on the previous day in three 0.5 gram doses (26 cases). [2] A single 4 gram dose was administered 30 minutes before treatment (30 cases). The results from the "excellent", "good" and "fair" cases showed a 16% increase in effectiveness when DPA was administered the day before, not a statistically significant difference (P less than 0.1), but a clear tendency to increase was observed. The above findings show that DPA has an enhancing effect on acupuncture analgesia and anesthesia in clinical practice.

Gomisin A (TJN-101) is one of the lignan components isolated from Schisandra Fruits and expected to have some efficacies in clinical treatment of hepatitis. The serum concentrations of TJN-101 and Met. B, which was identified as a demethylenated substance and one of the major metabolites of TJN-101 in rats, were investigated. After intravenous administration at doses of 1.6, 4.0 and 10 mg/kg of body weight, the serum concentration of TJN-101 decreased biphasically, and the terminal elimination half-life at each dose was about 70 min. Dose-dependency was observed for the area under the concentration-time curve (AUC). On the other hand, the serum concentration of TJN-101 increased rapidly and reached maximum within 15 to 30 min when administered orally. This result was supported by the in situ roop method. The Cmax and the AUC values were not exactly dose-dependent, but the values increased with a dose-up of TJN-101. The biotransformation of TJN-101 to Met. B, was very rapid in both intravenous and oral administrations. The AUC value of Met. B after oral administration of TJN-101 at a dose of 1.6 mg/kg was relatively larger than any other dosages. It suggested that TJN-101 was extensively underwent the first pass effect in rats. More than 80% of TJN-101 was bound with rat serum protein in vitro and in vivo. Therefore, it seems to be necessary to pay attention when it was administered concurrently with high protein binding drugs.

After oral administration of gomisin A (1) to rats, the bile and urine were collected and treated with beta-glucuronidase and arylsulfatase. Seven metabolites, met B (2), met A-III (3), met E (4), met D (5), met F (6), met G (7), and met H (8) were isolated from the bile treated with the enzymes. Eight metabolites 2-8, and met A-II (9) were isolated from the urine treated with the enzymes. A major metabolite 2, and two minor metabolites 3 and 9 were identified as met B, met A-III, and met A-II, respectively, which are oxidative products of 1 formed by rat liver S9 mix. The structures of five new metabolites 4-7, and 8 were determined on the basis of chemical and spectral studies.

正常ラットにおける肝部分切除による再生肝，肝組織血流量ならびに肝微細構造におよぼすTJN-101((+)-(6s，7s，R-biar)-5，6，7，8-tetrahydro-1，2，3，12-tetramethoxy-6，7-dimethyl-10，11-methylene-dioxy-6-dibenzo[a，c]cyclooctenol)の作用について検討した．1)TJN-101は肝部分切除後4日間の経口投与(10，30および100mg/kg/day)により肝再生率を上昇させるとともに，低下したBSP排泄能を改善した．これらの変化は用量依存的であり，100mg/kg/day投与群では擬手術群のレベルにまで回復していた．この際，TJN-101投与による血中生化学的指標の著明な変化として，肝切除により低下する総蛋白量の改善，LCAT活性の上昇，インスリンレベルの低下およびグルカゴンレベルの増加などが用量に依存して認められた．一方，肝細胞核分裂に対する変化は術後5日目ではほとんど認められなかった．2)TJN-101は30および100mg/kgの十二指腸内投与により肝組織血流量を増加させた．3)TJN-101(30，100および300mg/kg/day)4日間経口投与後の肝微細構造を電子顕微鏡にて検討したところ，特徴的な変化として100および300mg/kg/day投与群で粗面小胞体ならびに滑面小胞体の増加が認められた．以上の結果から，TJN-101は再生肝の肝細胞増殖ならびに肝機能の回復を促進させるとともに，肝内血流循環動態を亢進させる作用を有するものと推測された．また，TJN-101を反復投与した際にみられる肝重量増加は微細構造的には小胞体の発達を伴うものと考えられた．

ハリの鎮痛効果を増強する物質とされるD-フェニルアラニン (D-phenylalanine) を, 抜歯術に前投薬として応用し, その効果を検討した。方法はハリ麻酔単独で行った対照群31例と, ハリ麻酔30分前にD-フェニルアラニンを, 前投薬した試験群9例の抜歯術におけるハリ麻酔効果を比較することにより行った。対照群, 試験群ともに, 抜去歯に合わせて選穴したツボに, 中国針を刺針してハリ麻酔器に接続し, 約30分の誘導時間を経て抜歯術を行った。そしてそのハリ麻酔効果を2群間で比較した。効果判定はスコア1から5に分類し, それを著効・有効・やや有効・無効に判定した。その結果, 対照群のハリ麻酔成功率32%に対し, D-フェニルアラニンを前投薬した試験群の方が, 78%とハリ麻酔効果がより優れていた。

The effect of anticonvulsants on pentylenetetrazol (PTZ)-induced EEG power spectrum changes was examined. When the minimum dose of PTZ (15 mg/kg) was administered intravenously twice, with an interval of 80 min, a clear EEG power spectrum change was observed after the second PTZ administration, irrespective of whether or not the first PTZ administration evoked marked EEG changes. We defined the values F/N, S/N and S/F. The value F/N, obtained by dividing the power spectrum area after the first PTZ administration by the normal power spectrum area, was 1.06 +/- 0.05 (mean +/- S.E.). The value S/N, obtained by dividing the power spectrum area after the second PTZ administration by the normal power spectrum area, was 2.00 +/- 0.21. The value S/F, obtained by dividing S/N by F/N, was 1.86 +/- 0.16. The S/F value was almost constant regardless of whether or not the first PTZ administration could evoke marked EEG changes. The effect of anticonvulsants was examined by S/F value changes, and 100 mg/kg of PHT completely inhibited the double PTZ effect. Phenobarbital, ethosuximide and sodium valproate also inhibited the double PTZ effect. Using the S/F value of the EEG power spectrum with minimum dose double PTZ administration, quantitative evaluation is possible for anticonvulsant drugs.

In order to disclose the effect of “KANPO-YAKU” on immune responses, we investigated the induction of anti-SRBC response (PFC assay) in the spleen cells of BDF1 female mice which received Tsumura-Juzentaihoto (TJ-48) . TJ-48 induced an augmentation of anti-SRBC response by oral administration once a day for 7 consecutive days in a dose dependent manner. The maximum induction of anti-SRBC response was seen by 7 days-administration of TJ-48 (2 g/kg/day) and no further increase was obtained by longer treatment. After withdrawing TJ-48, the augmentation of anti-SRBC response was maintained for 3 days and disappeared by 7 days after the withdrawal. The anti-SRBC response was enhanced whether immunization with SRBC was done in vivo or in vitro.

Berberine has been shown to increase developed tension in cardiac muscle but its derivatives have been reported to inhibit the catalytic subunit of adenylate cyclase. In the present study, the cardiac actions of the most potent derivative, 13-propylberberine, were examined. It produced a transient increase followed by a sustained decrease in developed tension in paced left atrial muscle preparations isolated from guinea-pig heart. In the presence of 13-propylberberine, isoproterenol caused only a transient increase in developed tension; marked desensitization to the positive inotropic effect of isoproterenol occured within 20 min. After washout of isoproterenol and an additional 15-min incubation in the presence of 13-propylberberine, the muscle lost its sensitivity to isoproterenol. Moreover, the positive inotropic effect of ouabain or effects of decrease or increase in extracellular Ca 2+ concentration on the force of muscle contraction were markedly attenuated. Isoproterenol-induced elevation of tissue cyclic AMP concentration was inhibited by 13-propylberberine; however, 13-propylberberine did not alter the basal cyclic AMP concentration and its effects on inotropic actions of ouabain or extracellular Ca 2+ appear unrelated to tissue cyclic AMP concentration.

The effects of wuweizisu C, a lignan component of schizandra fruits, on liver injuries induced by carbon tetrachloride (CCl4), d-galactosamine and dl-ethionine were investigated by means of serum-biochemical and histopathological examinations in rats. Pretreatment or combined administration of wuweizisu C dose-dependently reduced the elevation of serum transaminase activity and histological changes such as fatty degeneration, cell necrosis, inflammatory cell infiltration, etc., which were caused by the single administration of 1 ml/kg, p.o., or the repeated administration of 0.2 ml/kg, s.c., daily for 4 days of CCl4, respectively. The effects of wuweizisu C on the liver injuries induced by a low dose (200 mg/kg, i.p.) and a high dose (400 mg/kg, i.p.) of d-galactosamine were compared with those of uridine. Wuweizisu C significantly lowered the rise of serum transaminase activity after the administration of a low dose of d-galactosamine in the serum-biochemical analysis. A tendency was also shown to inhibit cell necrosis and inflammatory cell infiltration caused by both doses of d-galactosamine in the histopathological examination. On the other hand, uridine markedly repaired the serum-biochemical and histopathological changes after the administrations of both doses of d-galactosamine. Also wuweizisu C cured the liver injury by the repeated administration of 150 mg/kg, i.p., daily for 4 days of d-galactosamine. After the repeated administration of 250 mg/kg, s.c., daily for 4 days of ethionine, liver cell atrophy, diffuse fatty degeneration and decrease of serum triglyceride were observed, but not cell necrosis. Wuweizisu C dose-dependently inhibited fatty degeneration and decrease of serum triglyceride. These findings suggest that wuweizisu C can be protective and/or therapeutic on hepatocellular phenomena such as cell necrosis, fatty degeneration, inflammatory cell infiltration, etc., in human hepatitis.

The effect of one of Kampo prescriptions, Tsumura Juzentaihoto (TJ-48), on phagocytes was investigated in BALB/c mice.The oral administration of TJ-48 for 3 to 5 days resulted in the enhanced phagocytic activity of peritoneal exudate cells (PEC) and bone marrow cells (BMC) and the maximum increase was observed at 2 g/kg/day of TJ-48. The enhanced phagocytosis was maintained for 5 days after the withdrawal of TJ-48 and returned to the basal level on day 7. A large increase of chemiluminescence was obtained in PEG from mice given TJ-48 orally for 7 days. Moreover, TJ-48 showed the direct activatingeffect on the phagocytosis of PEC and BMC in in vitro assay.

われわれは, さきに, D-フェニルアラニン (DPA) が, 鍼鎮痛効果を増強することを疼痛閾値の上昇より実験的に立証したことを報告してきた。今回は, 当麻酔科外来での慢性腰下肢痛患者を対象にDPAの術前投与法の違いによる効果を検討した。すなわち, 第I法は前日投与群 (1.5g 分3), 第II法は当日投与群 (4g) とし, これを偽薬投与群の設定により, 二重盲検法にて検討した。対象は罹病期間が3ヵ月以上の慢性腰下肢痛56例で, DPAの投与方法を2つに分けた。その (1) は, 1.5gを3回 (鍼治療前日の夕食後0.5g, 就寝前0.5g, 治療当日の朝食後0.5g) に分けて行い, 投与方法 (2) は, 4g (鍼治療前30分) を経口投与した。鍼治療は低周波置鍼療法を用いた。26例に対しては, (1) の投与方法で行い, 30例に対しては (2) の投与方法で行った。各々について「DPA+鍼」を2回,「偽薬+鍼」を2回, 計4回行い「直後効果」を数値スケール法によって両群を比較した。一方, ボランティア3名にDPA 4g・1.5gを内服させ, 2時間後, 4時間後及び翌月の血漿フェニルアラニンの経時的変動を調べた。その結果, 投与方法 (1) では,「DPA+鍼」(26例) 著効7.8%, 有効69.2%, やや有効19.2%であり (偽薬群に比べて有意), 投与方法 (2) によれば,「DPA+鍼」(30例) 著効23%, 有効37%, やや有効20%, 無効20%であった (偽薬群に比べて有意)。投与方法を比較すると, (2) より (1) の方が, 著効および有効を合わせると17%の増強を認めた。血漿フェニルアラニンの変動は4gの場合2時間～4時間後に高値を示し1.5gの場合は2時間後に高値を示した。われわれは, DPAの投与方法 (投与時期と投与分量) に関して臨床的な検討を行った。すなわち, 投与 (1) の方法と, 投与 (2) の方法との効果を比較すると, 慢性腰下肢痛における鍼鎮痛効果は, 当日投与よりも前日投与の方が, より効果的である傾向がわかった。症例数が少ないために再検討の余地はあるが, アメリカのエーレンプライスの提唱どおりDPAが slow onset, long acting に効果を発揮することに鑑み, 前日から分割して投与する方が望ましいと言える。

It has been demonstrated that TJ-8007 (Tsumura-Zokumeito, A traditional Chinese medicine) has a protective effect against cerebral anoxia. This study was done to elucidate the protective mechanism of TJ-8007 against cerebral ischemia and anoxia. TJ-8007 (0.3 approximately 3.0 g/kg, p.o.) inhibited the rise in the cumulative mortality rate after ligation of the bilateral carotid artery (BCA) in mice. TJ-8007 also significantly prolonged the survival time at the dose of 3.0 g/kg, p.o. However, TJ-8007 (1.0 or 3.0 g/kg, p.o.) did not affect the mean survival time after ligation of BCA in Mongolian gerbils and the gasping movement in a decapitated mouse head that served as a complete ischemic model. Ifenprodil (30 mg/kg, p.o.) also showed the protective effect only against ischemic death after ligation of BCA in mice. TJ-8007 (1.0 or 3.0 g/kg, p.o.) increased the vertebral blood flow, but showed no effect on the internal carotid blood flow in anesthetized dogs. These results suggest that the mechanism for the cerebral protective effect of TJ-8007 may be due to its ameliorating action on the cerebral circulation.

Physical dependence liability of chlorphenesin carbamate (CPC) was studied in parallel with phenobarbital-Na (PB). Beagle dogs were used and the overall duration of the experiment was 85 days, i.e. the first dosing period was 42 dyas (6 weeks) in which drugs were repeatedly administered orally once daily, followed by a withdrawal period (7 days), the second dosing period was continued from the 50th-78th day in which the form and schedule of drug administration was as in the first dosing period. The last 79th to 85th days were used for substitution experiments. In both dosing periods, PB but not CPC showed signs of tolerance formation. Severe withdrawal syndrome was observed in PB administered dogs whereas there were no changes of behavior observed in CPC-dogs by withdrawal and substitution procedures, respectively. CPC apparently does not have a physical dependence liability.

Δ1-Tetrahydrocannabinol (Δ1-THC, Ia) was oxidized with selenium dioxide to yield 7-hydroxy-(VI), 6α-hydroxy-(VIIa), 6β-hydroxy-(VIIIa) and 6-oxo-Δ1-THC (IX) together with cannabinol (CBN, III), 7-hydroxy-(X), 6-hydroxy-(XI) and 7-oxo-CBN (XII). Δ6-THC (IIa) was transformed merely to 7-hydroxy-(IVa) and 7-oxo-Δ6-THC (Va) in a similar manner. Individual products isolated are well defined mainly on the spectroscopic evidences. The oxidation of Ia favors the endocyclic allyl methylene rather than the exocyclic allyl methyl while the reverse is the case with IIa. Such a sharp contrast shown in their reactivities as mentioned above must be interpreted in terms of the steric interference at the reactive center.

Experiments were performed to investigate the effects of TJ-41 on spermatogenic disorders under current treatment with adriamycin (ADR). Male ICR mice were intraperitoneally injected with ADR at the dose of 0.15 mg/kg, twice a week for 5 weeks. Simultaneously, these mice were orally administered TJ-41 at the dose of 1, 2 or 4 g/kg for 12 weeks. The effects of TJ-41 were evaluated by histological analysis of germ cells in the testis at 7 weeks after the last injection of ADR. TJ-41 at a dose of 4 g/kg significantly inhibited the decrease of testis weight in mice treated with ADR. TJ-41 at doses of 1 and 4 g/kg significantly decreased the proportion of seminiferous tubules without germ cells as compared with the ADR-treated group. On the other hand, TJ-41 at doses of 1 and 4 g/kg significantly increased the proportion of normal seminiferous tubules and the Sertoli cell ratio of spermatocytes as compared with the ADR-treated group. These results indicate that TJ-41 may qualitatively and quantitatively protect against the decrease of germ cells in the testis of mice treated with ADR.

The s.c. administration of 20 mg/kg of morphine-HCl produced a decrease in the spontaneous locomotor activity (SLMA) of rats. The decrease in SLMA was significantly antagonized by p-chlorophenylalanine (p-CPA). When rats pretreated with p-CPA were given 5-hydroxytryptophan before morphine injection, the marked sedative response to morphine was restored, suggesting that the morphine-induced decrease in SLMA of rats may depend on the release of 5-hydroxytryptamine by morphine. By contrast, the s.c. administration of 5 mg/kg of morphine-HCl produced a significant increase in SLMA of rats. The magnitude of the increase was reduced by atropine, scopolamine or alpha-methyl-p-tyrosine. It appears that both adrenergic and cholinergic mechanisms participate in the increase in SLMA of rats induced by morphine. Both the increase in SLMA produced by 5 mg/kg of morphine and the decrease in SLMA induced by 20 mg/kg of morphine were completely antagonized by the s.c. administration of naloxone-HCl, 0.0625 and 0.25 mg/kg, respectively. Thus, it appears that the receptor with which morphine interacts to produce stimulation is chemically identical with or very similar to the receptor with which morphine combines to induce depression. The former receptors, however, are likely to be located on different neurons from the latter.

TJN-101 ((+)-(6S,7S,R-biar)-5,6,7,8-tetrahydro-1,2,3,12-tetramethoxy -6,7-dimethyl-10,11-methylenedioxy-6-dibenzo[a,c]cyclooctenol) is one of the lignan compounds isolated from Schisandra fruits. 1) Effect of TJN-101 on liver fibrosis was investigated in rats which were injected with CCl4 (1 ml/kg) subcutaneously twice a week for 12 weeks. TJN-101 was given orally at the dose of 10 or 30 mg/kg/day for 6 or 3 weeks beginning on the 6th or 9th week after the start of CCl4-intoxication, respectively. The elevations of serum transaminase activities and the increase of liver 4-hydroxyproline content were observed depending on the period of CCl4-intoxication. These changes were marked on the 9th and 12th weeks after. In the histopathological study, the degenerative fatty change on the 6th week after and the formation of pseudolobule caused by fibrosis proliferation on the 9th or 12th week after were mainly observed. When rats were treated with TJN-101, the abnormalities in biochemical parameters and the fibrosis proliferation caused by CCl4-intoxication were improved. 2) Chronic liver injury was induced by the treatment with CCl4 (1 ml/kg) subcutaneously twice a week for 10 weeks to investigate the effect of TJN-101 on liver regeneration after partial hepatectomy. TJN-101, which was given orally at the dose of 10, 30 or 100 mg/kg/day for 6 days from the 1st day after partial hepatectomy, dose-dependently increased the liver regeneration rate and improved the serum BSP retention rate. These results suggest that TJN-101 suppresses the fibrosis proliferation and accelerates both the liver regeneration and the recovery of liver function after partial hepatectomy in chronic liver injury.

Rat epidermal growth factor (rEGF) was isolated from the submaxillary gland of male rat by reversed phase high performance liquid chromatography and ion exchange chromatography. The binding of purified rEGF to human carcinoma cells (A-431) and its tritiated thymidine uptake on rat epidermal fibroblast cells (FR) were almost the same as those of purified commercially available mouse EGF (mEGF). Antisera to rEGF was raised in rabbits and a radioimmunoassay (RIA) system was established. The assay range of the RIA was about 1.0 to 100 ng/ml. The within assay coefficients of variation were 5 to 10%, while the between assay coefficients of variation were 5 to 13%. The tissue content of rEGF of male rats (10 weeks old) was examined. As a result, the submaxillary gland was found to contain a very high concentration of rEGF (214 micrograms/g wet tissue) as predicted, and digestive tissues, stomach, intestine and duodenum contained 2.49, 3.57, 9.44 ng/g wet tissue, respectively. The amounts in prostate and seminal vesicle were relatively high, being 65.6 and 2,268 ng/g wet tissue, respectively. The amount in the submaxillary gland increased markedly after 7 weeks of age. These results suggest that EGF is an important factor in gonadal function.

The mitogenic activity of TJ-48 in murine lymphoid cells was examined to get a clue for the augmentation of antibody production by TJ-48.Mitogenic activity of TJ-48 was detected in spleen cells, lymphonodus cells but not in thymocytes. The effect was abolished by pretreatment with anti-Ig antibody but not affected by pretreatment with anti-Thy 1.2 antibody. Coculture of spleen cells with TJ-48 resulted in increased number of sIgM-, sIgG- or sIgD-positive cells according to FAGS analysis. The increase in mitosis by TJ-48 was abolished by the elimination of adherent cells and the readdition of adherent cells recovered the effect of TJ-48. These data indicate that TJ-48 is a B cell mitogen and the activity is T cell-independent and adherent cell-dependent. The mitogenic activity of TJ-48 may account for the augmentation of the immune response.

Adult male rats were administered morphine twice a day for 45 days and the effects of morphine on the copulation rate, the weight of various organs, and on the hypothalamic-pituitary-gonadal axis were examined. Morphine administered rats showed a loss of weight, hypertrophy of the adrenals, decreased weight of accessory sex organs, low sperm count, and decreased copulation rate. The contents of the luteinizing hormone releasing hormone in the hypothalamus and the luteinizing hormone in the pituitary remained unchanged. Serum luteinizing hormone and testosterone levels decreased, but serum follicle-stimulating hormone levels increased. These results suggest that morphine inhibits the hypothalamic-pituitary-gonadal axis and causes a diminution in the number of fertilizations of the partner females.

To elucidate the differences between neurons of epileptogenic animals and those of normal animals, cellular characteristics of neurons of mutant strain El mice which are highly susceptible to seizures were investigated using immunocytochemical techniques. In neurons of 3-day primary cultures, the control ddY mouse neurons showed dividing stages in about 0.2% of neurofilament (NF)-positive neurons, whereas no dividing neurons were observed among the NF-positive El mouse neurons. In 7-day cultures, localization of GD3 ganglioside in the proliferating control ddY mouse neurons was observed, but there was no GD3 ganglioside in the mutant El mouse neuron. The content of GD3 ganglioside detected by high-performance thin-layer chromatography of El mouse cultured cells was ca. 1/4 of that of ddy mice. These findings suggest that neurons of the El mouse are differentiated earlier than those of the control ddY mouse.

Introduction:There are two types of phenylalanine, one of the essential amino acids, D-Type and L-type. Phenylalanine in itself has shown no clear analgesic effect in human beings, however, as we reported previously, D-type phenylalanine (DPA) when administered prior to acupuncture analgesia in human beings lengthens the analgesic effects of acupuncture. At this time then, using volunteers in whom acupuncture analgesia failed to raise the pain threshold (PT) we experimentally studied whether or not DPA has any influence.Methods:Subjects were divided into an acupuncture anesthesia effective group (a rise in PT was observed due to acupuncture) and an acpuncture anesthesia ineffective group (no rise in PT). 4.0g. DPA was administered orally in both groups 30 minutes prior to acupuncture anesthesia. For acupuncture anesthesia right and left LI-4 and right and left ST-36 were used with low frequency electric current administered for 50 minutes. For the determination of PT a radiant heat pain meter was used. The PT was measured when DPA was administered, during acupuncture anesthesia and at every 30 minutes for 3 hours after the needles were withdrawn.Results:In the acupuncture anesthesia effective group, a rise in the PT was observed in 4 of the 5 cases and, the rise was much faster than when acupuncture anesthesia was used alone. In the acupuncture anethesia ineffective group, there were cases in which a remarkable rise in PT was observed or in which no rise in the PT occured during acupuncture stimulation but occured after the discontinuation of the acupuncture and some cases in which no rise occured during or after anesthesia.Discussion:1. In the acupuncture anesthesia effective group due to DPA pre-medication, the rate of PT rise quickened.2. After operations the PT rise was lengthened.3. The individuality of the acupuncture effectiveness in human beings was partially eliminated.

A rapid and sensitive method for determination of morphine in human urine using gas chromatography is described. The method involves charcoal absorption and solvent extraction of morphine followed by gas chromatographic analysis of the trimethylsilyl derivative. It is possible to detect morphine by this method more than 96 hrs after injection.

Either pethidine HCl (50 mg/kg s.c.) or methadone HCl (8 mg/kg s.c.) produced a prominent decrease in locomotor activity of rats. Pretreatment of rats with p-chlorophenylalanine (p-CPA, 320 mg/kg i.p.) 48 h before the narcotic injection significantly antagonized the activity-decreasing effects of narcotics. When rats pretreated with p-CPA were given 5-hydroxytryptophan (75 mg/kg s.c.) 30 min before narcotic administration, the activity-decreasing response to narcotics was restored. Thus, a decrease in locomotor activity induced in rats by either pethidine or methadone is probably mediated by serotonergic mechanisms.

Pharmacological actions of (6)-shogaol and capsaicin were studied. Both (6)-shogaol (0.5 mg/kg, i.v.) and capsaicin (0.1 mg/kg, i.v.) caused a triad such as a rapid fall in blood pressure, bradycardia and aponea in rats. Both drugs-induced marked pressor responses in blood pressure, which occurred after the rapid fall, were markedly reduced by a spinal destruction. In pithed rats, both drugs-induced peripheral pressor responses were markedly reduced with the combined treatment of [D-Arg1, D-Pro2, D-Trp7,9, Leu11]-substance P (0.5 mg/kg, i.v.), phentolamine (10 mg/kg, i.v.) and the section of sciatic nerves. In isolated guinea-pig trachea, (6)-shogaol (100 microM) and capsaicin (10 microM) induced contractile responses which were slightly inhibited by substance P antagonist (10 microM), but exhibited also a tachyphylaxis. Furthermore, although (6)-shogaol (3.6 microM) showed positive inotropic and chronotropic actions on isolated atria in rats, this effect of (6)-shogaol disappeared by repeated injections or pretreatment (100 mg/kg, s.c.) of (6)-shogaol. These results suggest that (6)-shogaol and capsaicin have similar actions, and that both drugs may cause a peripheral action by releasing an unknown active substance from nerve ends.

The radioimmunoassay for methamphetamine (I) in urine of man has been established by use of the 125I-labelled derivative of N-[3-(p-hydroxyphenylacetylamino) propyl]-methamphetamine (V) in sensitivity of 8.0 ng/100μl. The antigen binding capacity of the antiserum prepared form N-carboxymethylmethamphetamine-BSA (II) was determined by a new method of immunoassay using fluorescence labelled N-(3-dansylaminopropyl) methamphetamine (IV) ("fluoroimmunoassay"). The specificity of the antibody was examined by its cross reaction with several methamphetamine analogues.

General pharmacological studies were performed on (6)-gingerol and (6)-shogaol which are the pungent constituents of ginger (Zingiber officinale Roscoe). Intravenous (i.v.) administration of (6)-gingerol (at 1.75-3.5 mg/kg) or (6)-shogaol (at 1.75-3.5 mg/kg) and oral administration of them (at 70-140 mg/kg) produced an inhibition of spontaneous motor activity, an antipyretic and analgesic effects, prolonged hexobarbital-induced sleeping time, and these effects of (6)-shogaol were mostly more intensive than that of (6)-gingerol. (6)-Shogaol showed an intense antitussive effect in comparison with dihydrocodeine phosphate. In the electro-encephalogram of cortex, the low amplitude fast wave pattern was observed for 5 min after i.v. administration of (6)-shogaol, and then changed to the drowsy pattern, which was restored after 60 min. In the gastro-intestinal system, (6)-shogaol intensively inhibited the traverse of charcoal meal through the intestine in contrast with (6)-gingerol after i.v. administration of 3.5 mg/kg, but (6)-shogaol facilitated such an intestinal function after oral administration of 35 mg/kg. Both (6)-shogaol and (6)-gingerol suppressed gastric contraction in situ, and the suppression by the former was more intensive than that by the latter. In the cardiovascular system, both (6)-shogaol and (6)-gingerol produced depressor response at lower doses on the blood pressure. At high doses, both drugs produced three phase pattern.