HLA Gene in 5 Cases of Liver Injury Related to Baikal Skullcap.

HLA association with drug-induced liver injury has recently been pointed out about multiple medicines. The aim of this study was to evaluate relationship between HLA gene and liver injury related to Baikal skullcap-containing Kampo medicines (BSCK). We previously examined HLA genes in 3 cases of BSCK-induced liver injury. Recently we could encounter 2 cases diagnosed as "definitely-related case" of BSCK-induced liver injury. HLA genes of the 2 cases were analyzed by Sequencing Based Typing method with Next Generation Sequencer at HLA Laboratory in Kyoto. HLA-DPA1*02:02:02 and DPB1*05:01:01 were observed in the 2 cases: concordance was not observed in HLA-A, B, C, DRB1, DRB4, DQA1, or DQB1. The previous 3 cases of BSCK-induced liver injury had the same allele type to the 2 cases only in HLA-DPA1. Putting all these together, HLA-DPA1*02:02:02 was observed in common among 5 cases of BSCK-induced liver injury. HLA-DPA1*02:02:02 is possibly associated with BSCK-induced liver injury.

A Study of Factors Associated with the Development of Pseudoaldosteronism in Outpatients.

Objective: The development of pseudoaldosteronism is shown to be mainly associated with four factors: daily dose of glycyrrhiza (licorice), duration of glycyrrhiza use, body size, and age. Recently, direct bilirubinemia and hypoalbuminemia are newly reported as possible factors that trigger pseudoaldosteronism due to glycyrrhiza ingestion. Pseudoaldosteronism occurs in the presence of combinations of these factors; therefore, the importance of each factor on the tolerance to glycyrrhiza loading is still unclear. Methods: In seven patients (63-78 years old, six women) who developed pseudoaldosteronism due to ingestion of glycyrrhiza-containing Kampo extract in their clinic, serum albumin and direct bilirubin (D-bil) levels were investigated. In six women, the authors evaluated the correlations between daily dose of glycyrrhiza ingested and each factor: age, height, weight, body mass index, body surface area (BSA), and duration of ingestion (Pearson's correlation coefficient). Results: No patients had abnormal levels of serum albumin or D-bil around the time of the onset. In six women, the highest correlation coefficient was observed between BSA and the glycyrrhiza dose in Kampo extract at the onset of pseudoaldosteronism. Conclusions: The findings suggested that in elderly women, BSA should be considered first as a factor for predicting the development of pseudoaldosteronism.

Macrophage-mediated inhibitory effect of Zingiber officinale Rosc, a traditional oriental herbal medicine, on the growth of influenza A/Aichi/2/68 virus.

The inhibitory effect of Zingiber officinale Rosc (ZOR), an Oriental traditional herbal medicine, on the growth of influenza A/Aichi/2/68 (Aichi) virus was investigated in Madin-Darby canine kidney (MDCK) cells. Direct addition of ZOR (0.1 approximately 100 microg/ml) to the infected cells did not have any inhibitory effect. However, the ZOR-induced conditioned medium (ZOR-CM) of RAW cells, a murine macrophage (Mphi) cell line, exhibited an apparent inhibitory effect on MDCK cells without cytotoxicity. In accordance with the time-dependent inhibitory effect of ZOR-CM, it has been demonstrated that tumor necrosis factor (TNF)-alpha was gradually accumulated in ZOR-CM by the induction of TNF-alpha mRNA expression in ZOR-stimulated RAW cells. Conversely, the inhibitory effect of ZOR-CM was reduced significantly by the removal of TNF-alpha after the formation of an immune complex with anti-TNF-alpha monoclonal antibody. These data suggested that ZOR itself has no inhibitory effect on the growth of influenza virus, but could exert its effect via macrophage activation leading to production of TNF-alpha.

Suppressive effects of Stephania tetrandra on the neutrophil function in patients with rheumatoid arthritis.

Crude preparations of Stephania tetrandra (ST), a traditional herbal medicine, have been used safely for arthritis and silicosis in China. The concentration of granulocyte elastase - alpha 1 protease inhibitor complex in plasma is enhanced in inflammatory processes, e.g. in septicaemia and rheumatoid arthritis (RA), being an expression of granulocyte activation during inflammatory response. It has previously been reported that ST showed beneficial and immunomodulatory effects in the treatment of relatively mild RA. After the administration of ST for 12 weeks, the proportion of granulocytes and the granulocyte count in peripheral blood decreased significantly. The lipid peroxide and human granulocyte elastase levels of stored plasma declined significantly. Furthermore, both the leukocyte/elastase ratio and granulocyte/elastase ratio increased significantly. The findings of this study suggest that the suppressive effect of ST administration on excessive granulocyte activation resulted in the improvement of inflammation with rheumatoid arthritis.

Inducible activity of ginger rhizome (Zingiber offifinale Rosc.) on the mRNA expression of macrophage-inducible nitric oxide (NO) synthase and NO production in a macrophage cell line, RAW264.7 cells.

We have investigated the effect of Zingiber offifinale Rosc. (ZOR) on macrophage-inducible nitric oxide (NO) synthase (macNOS) mRNA expression and NO production in RAW264.7 cells, a murine macrophage cell line; 100 microg/ml ZOR can induce macNOS mRNA expression, but induction effects at a dose below 10 microg/ml were weak or negligible. Kinetic studies showed that macNOS mRNA can be detected from 4 hours to 24 hours after dosing, with a peak at 8 hours. In accordance with the induction of macNOS mRNA expression, NO concentrations increased from 3.4 microM at 2 hours to almost 150 microM at 24 hours, reflecting a longer period of macNOS mRNA expression. The activity of ZOR can be considered to contribute, at least in part, to the beneficial effects of ZOR through the macNOS-mediated activation of the biodefense mechanism.

Challenge tests and kampo medicines: case report and review of the literature.

In order to diagnose Kampo medicine-induced liver injury, a challenge test using the suspected medicine is the most reliable method of assessing the relationship between Kampo medication and liver injury. However, such a challenge test may cause severe liver injury. We examined the clinical features and safety of challenge tests conducted by a physician in a case of Kampo medicine-induced liver injury that we encountered as well as in the previous literature (six cases) in Japan. In all cases except two, one-third of the daily dose was given a few times for the challenge test (challenge dose was not described in two cases). The reaction induced by the challenge peaked 1-3 days after challenge. Mild eosinophilia (6%) was observed in two cases. All liver injuries induced by challenge cleared within 2 weeks, and neither severe nor fatal liver injury was observed. In conclusion, a challenge with a small dose of Kampo medicine conducted in our case and in the previous literature induced mild and reversible liver injury. The safety and availability of challenge with a small dose of Kampo medicine should be further examined in a larger population with Kampo medicine-induced liver injury.

Reexamination of the relation between menstrual cycle and kampo diagnosis, yin-yang.

Several Chinese herbal medicine textbooks describe that a short menstrual cycle indicates "yang" status and a long cycle indicates "yin" status. However, we sometimes encounter yang patients with a long cycle or yin patients with a short cycle in daily practice. Therefore, we reviewed the relationship between yin-yang and length of menstrual cycle. A questionnaire addressing both menstrual cycle and several parameters for determining yin-yang was completed at the time of the patients' visit to the hospital. Patients were divided into yang or yin groups, and menstrual cycle was compared between the two groups. A short menstrual cycle was mostly observed in yin patients (27.42 +/- 1.73, 95% CI: 26.32-28.52, n = 12) and a long cycle was mostly observed in yang patients (31.63 +/- 3.96, 95% C.I: 29.51-33.74, n = 16). The difference in menstrual cycle between the two groups was significant (p = 0.002), and this result was entirely opposite to the traditional theory. This study identified a discrepancy between the traditional theory and the actual menstrual cycle findings in this small population. Current reexamination of the relationship between yin-yang and menstruation in a larger population is warranted.

Lymphocyte transformation test for medicinal herbs yields false-positive results for first-visit patients.

We performed lymphocyte transformation tests (LTTs) for toki (angelicae radix) and ogon (scutellariae radix) on first-visit patients who had never taken Kampo medicines. LTTs for both herbs were positive in 12 of 14 patients, suggesting that LTTs for these herbs are unreliable for the diagnosis of Kampo medicine-induced liver injury.

Four cases of panic disorder successfully treated with Kampo (Japanese herbal) medicines: Kami-shoyo-san and Hange-koboku-to.

Kampo medicines have been used to treat patients with psychogenic disorders from ancient times. In the present report the cases are described of four patients with panic disorder successfully treated with Kampo medicines. These four patients fulfilled the Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria for panic disorder with agoraphobia. The Kampo medicine Kami-shoyo-san (TJ-24) relieved panic attacks, anticipatory anxiety and agoraphobia in two patients, and Hange-koboku-to (TJ-16) relieved these symptoms in the other two patients. The patients in whom Kami-shoyo-san was effective were older and complained of more symptoms than those in whom Hange-koboku-to was effective. These Kampo medicines may be useful as additional or alternative treatments for panic disorder.

The efficacy of herbal medicine (kampo) in reducing the adverse effects of IFN-beta in chronic hepatitis C.

The purpose of this study was to determine if the adverse effects of interferon (IFN) in hepatitis C patients could be reduced by treatment with Japanese Oriental (Kampo) medicine. Twelve patients with chronic hepatitis C were treated with a combination of IFN-beta and either Mao-to or Dai-seiryu-to (groups A and B), and 16 patients were treated with IFN-beta alone (group C). Mao-to was administered to eight patients and Dai-seiryu-to was administered to four in groups A and B, respectively. Adverse effects were evaluated by clinical and laboratory examinations. The severity of symptoms was daily self-classified into four categories (1: none, 2: very slight, 3: moderate, and 4: serious), using a questionnaire consisting of 29 items. Scores of symptom such as discomfort and fever in group A, and discomfort, general malaise, paresthesia and arthralgia in group B were significantly lower than those in group C (p < 0.05). In all patients, HCV-RNA was negative at the end of the treatment, and serum alanine aminotransferase (ALT) levels had normalized transiently in all group A and B patients with genotype 1b by 2 weeks after cessation of IFN treatment. This study indicates that Kampo medicines are useful for reducing the adverse effects accompanying IFN treatment in patients with chronic hepatitis C without reducing the antiviral effects.

Additional inhibitory effect of tea extract on the growth of influenza A and B viruses in MDCK cells.

It has been previously reported that green-tea extract (GTE) inhibits the growth of influenza virus by preventing its adsorption. In this study, we further investigated whether GTE exerts an additional inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS) and thereby inhibits the growth of influenza A and B viruses in Madin-Darby canine kidney cells. The vital fluorescence microscopic study showed that GTE inhibited acidification of ELS in a concentration-dependent manner. Moreover, the growth of influenza A and B viruses was equally inhibited when the cells were treated with GTE within as early as 5 to 15 min after infection, depending on the virus strains. The fact that (-)epigallocatechin (EGC), one of major catechin molecules in GTE, exerts the inhibitory effects on the acidification of ELS and virus growth in a manner similar to that of GTE strongly suggests that EGC is one of the active components in the extract.

Herbal medicine and false-positive results on lymphocyte transformation test.

In vitro mitogenic activity of 16 herbs and 3 Kampo (herbal medicine) formulae have been reported in experimental studies. It is not known how many herbs and Kampo formulae in total have mitogenic activity. Lymphocyte transformation test (LTT) is generally utilized to diagnose drug-induced liver injury. In LTT, mitogenic activity is assessed by measuring 3H-thymidine incorporation. The objective of the present study was to determine which herbs and which Kampo formulae caused false-positivity on LTT. We examined 2496 summaries of all admission records from 1979 to 1999 in our department. We selected patients in whom liver injuries were diagnosed as definitely unrelated to Kampo medication. In these patients, LTT was performed for some herbs contained in the suspect Kampo medicines, resulting in positive LTT for 17 herbs: Evodiae Fructus (Goshuyu), Zizyphi Fructus (Taiso), Ginseng Radix (Ninjin), Zingiberis Rhizoma (Shokyo), Hoelen (Bukuryo), Aconiti Tuber (Bushi), Angelicae Radix (Toki), Cnidii Rhizoma (Senkyu), Rehmanniae Radix (Jio), Ephedrae Herba (Mao), Anemarrhenae Rhizoma (Chimo), Cinnamomi Cortex (Keihi), Bupleuri Radix (Saiko), Artemisiae Capillari Spica (Inchinko), Persicae Semen (Tonin), Moutan Cortex (Botanpi) and Paeoniae Radix (Shakuyaku). These results were considered false-positive, because the results were observed in the "definitely unrelated" patients. Mitogenic activity inherent to some herbs and Kampo formulae may sometimes cause false-positivity on LTT in clinical situations. These examples suggest that LTT for Kampo formulae may be unreliable as a diagnostic method for drug-induced liver injury.

Rhubarb use in patients treated with Kampo medicines--a risk for gastric cancer?

In vitro mutagenic effects have been reported for ingredients contained in rhubarb. Therefore, rhubarb (Rhei Rhizoma) as an anthranoid laxative could be associated with a risk of developing gastric cancer as well as colorectal cancer. We are not aware of any reports that have examined the relationship between the use of rhubarb and the development of gastric cancer. During the period between 1979 and 1999, we treated 14,616 patients using various Kampo medicines, which sometimes contained rhubarb. In the present study, we determined whether patients, diagnosed with gastric cancer during the period between 1979 and 1999, had been administered rhubarb before the development of gastric cancer. Among the 10 enrolled patients, only 2 patients had been administered rhubarb before the development of gastric carcinoma. The other 8 patients had never received rhubarb before the development of gastric carcinoma. Rhubarb use may have little connection with the development of gastric cancer in practice, even if some ingredients in rhubarb have shown carcinogenic activity in experimental studies.