Identification of a predictive biomarker for the beneficial effect of keishibukuryogan, a kampo (Japanese traditional) medicine, on patients with climacteric syndrome.

Keishibukuryogan (KBG; Guizhi-Fuling-Wan in Chinese) is one of the Kampo (Japanese traditional) medicines used to treat patients with climacteric syndrome. KBG can be used by patients who cannot undergo hormone replacement therapy due to a history of breast cancer. We evaluated whether cytosine-adenine (CA) repeat polymorphism of the estrogen receptor ß gene can be a predictor of the beneficial effect of KBG on climacteric syndrome. We also investigated the relationship between CA repeat polymorphism, the patients' profiles, and the therapeutic effect. We found that CA was an SS, SL, or LL genotype according to the number of repeats. We studied 39 consecutive patients with climacteric disorders who took KBG for 12 weeks. The diagnosis of climacteric disorders was made on the basis of the Kupperman index. KBG significantly improved the patients' climacteric symptoms (i.e., vasomotor symptoms in the patients with the LL genotype and melancholia in the patients with the SL genotype). No relationship between the patients' profiles and CA repeat polymorphism was recognized. CA repeat polymorphism could thus be a potential biomarker to predict the efficacy of KBG in climacteric syndrome, and its use will help to reduce the cost of treating this syndrome by focusing the administration of KBG on those most likely to benefit from it.

Dietary agonists of TRPV1 inhibit gastric acid secretion in mice.

Capsaicin and 6-gingerol, pungent components of chilli pepper and ginger, are known as dietary agonists of transient receptor potential vanilloid-1. Transient receptor potential vanilloid-1 nerve fibers are recognized to play a role in gastric mucosal integrity in rats. In the present studies, we examined the acute effects of peroral administration of capsaicin and 6-gingerol on gastric acid secretion in conscious mice. These agents were given p. o. 30 min before the pylorus was ligated. Oral administration of capsaicin (1.0-100 mg/kg) or 6-gingerol (1.5-50 mg/kg) significantly and dose-dependently inhibited basal acid secretion. Pretreatment with BCTC, a transient receptor potential vanilloid-1 antagonist, significantly reversed the reduced basal acid secretion by capsaicin or 6-gingerol. The combination of the lowest doses of capsaicin and 6-gingerol markedly inhibited basal acid secretion in conscious mice and this was also significantly reversed by BCTC. Moreover, the combination of the maximal dose of capsaicin and 6-gingerol inhibited basal acid secretion only to the level of a single administration of the maximal dose of capsaicin. These results suggest that the combination of capsaicin and 6-gingerol has an additive effect on the inhibition of gastric acid secretion through activation of transient receptor potential vanilloid-1. In separate experiments, intraduodenal administration of either capsaicin (30 mg/kg) or 6-gingerol (15 mg/kg), whose doses were observed to have a significant inhibitory effect by oral administration, tended to inhibit basal acid secretion compared with the vehicle. These results suggest that the combination of capsaicin and 6-gingerol has an additive effect on inhibition of gastric acid secretion through activation of transient receptor potential vanilloid-1, and oral administration of transient receptor potential vanilloid-1 agonists directly stimulates transient receptor potential vanilloid-1 in the gastric lumen, resulting in a potent reduction of gastric acid secretion.

Analysis of body constitution of fifty-two patients with Stevens-Johnson Syndrome (SJS) using Kampo Medical Questionnaires: prediction of SJS based on body constitution using decision tree.

UNLABELLED: Reports have indicated a relationship between adverse drug reaction (ADR) and Human Leukocyte Antigen (HLA) polymorphism and a relationship between Body Constitution (BC) and HLA polymorphism. Thus, a relationship between ADR and BC is suggested. We therefore created a questionnaire (hereinafter "Questionnaire") to survey the typical BC of Stevens-Johnson Syndrome (SJS) patients to determine how they differ from healthy persons, and studied the relationship between the development of SJS and BC. The Questionnaire had 30 typical items selected from those relevant to the BC necessary for the diagnosis and therapy of Sho-syndrome in Kampo Medicine. In the comparison of the prevalence of BCs between SJS patients and control persons, the prevalence of three BCs in the SJS group was significantly higher than that in the control group: 1) Does your throat ever feel closed up? ANSWER: Yes, 2) Do you easily feel hot flashes or burning cheeks even though your hands and feet feel cold? ANSWER: Yes, and 3) Do your lips or gums look dull red? ANSWER: Yes. In the analysis using the decision tree, the concentrated group of SJS patients (eighty-fold) was extracted using two decision trees consisting of 3 index variables. Persons with BCs from any of 1) to 3) are suggested to be at high risk of developing SJS.

A case of thalamic pain successfully treated with Kampo medicine.

OBJECTIVE: Thalamic pain, one of the central painful poststroke symptoms, is a severe pain that is often intractable. A case of thalamic pain successfully treated with Kampo medicine is presented. SUBJECT: A 65-year-old woman complained of moderate continuous and paroxysmal severe pain in the right upper and lower limbs after she had suffered from stroke. She also complained of sensory disturbance and intermittent involuntary movement. A magnetic resonance imaging scan of the brain showed an old infarction in the left thalamus. INTERVENTIONS AND OUTCOME: Paroxetine was administered, but it was stopped because of nausea. Etizolam was effective in reducing the pain for only about 30 minutes. Sokeikakketsuto decoction, one of the Kampo medicines, was administered orally on the basis of Kampo diagnostic criteria. Ten (10) days later, the pain had almost disappeared, and the other symptoms had also improved. CONCLUSIONS: This result suggested that Sokeikakketsuto could be an option for the treatment of thalamic pain under certain conditions.

A case of mediastinal lymphangioma successfully treated with Kampo medicine.

OBJECTIVE: The authors report a case of mediastinal lymphangioma successfully treated with Kampo medicine. METHODS: A 2-year-old boy with an axillary soft mass consulted our clinic. Physical examination findings were normal except for axillary elastic swelling. The neck and chest magnetic resonance imaging scan (MRI) showed a multilocular mass starting from a cervical lesion and extending above the carina. RESULTS: After 9 months of Kampo administration, MRI showed marked regression of mediastinal lymphangioma. CONCLUSIONS: It was found that Kampo medicine might be safe and effective as an alternative choice of treatment for lymphangiomas.

A suspected case of somatoform disorder successfully treated with an herbal medicine.

OBJECTIVE: Subjective physical symptoms, irrespective of whether they are psychosomatic or not, do not always show obvious or reasonable signs in examinations, which often makes the differential diagnosis between somatoform disorders and actual physical disease difficult for psychiatrists. In addition, psychiatrists have few clues as to how to treat diverse "medically unexplained" symptoms. This difficulty has highlighted the need for alternative treatments for somatoform disorders. SUBJECT: A 16-year-old high school baseball player was suffering from coxalgia and was unable to walk without crutches over 6 months. No painkiller was effective, the orthopedist found no remarkable signs in any examinations, and the patient was psychiatrically diagnosed with undifferentiated somatoform disorder. However, conventional therapies such as psychotherapy and selective serotonin reuptake inhibitors were ineffective. INTERVENTIONS AND OUTCOME: The therapeutic strategy was reevaluated from the perspective of Kampo diagnostics and keishikajutsubuto, a traditional Japanese herbal (Kampo) medicine, was chosen to be prescribed, which had a remarkable effect. His leg function improved within 2 weeks, and his pain and need for crutches disappeared in 6 weeks. CONCLUSIONS: Keishikajutsubuto has a different pain-relieving effect from conventional therapies. Kampo medicine thus provides an alternative approach for treating medically unexplained symptoms without strictly distinguishing between physically existing illness and psychologically caused somatoform disorders. Although details regarding the therapeutic mechanisms of Kampo medicine remain unclear and further studies are needed to increase its usefulness in clinical practice, Kampo medicine should be considered as an alternative treatment, especially for somatoform disorders.

Principal component vector rotation of the tongue color spectrum to predict "Mibyou" (disease-oriented state).

PURPOSE: Kampo medicine (Japanese traditional herbal medicine) contains concepts useful for preventive medicine. For example, "Mibyou" (disease-oriented state) aims to prevent illness by early recognition. Kampo diagnosis is based on subjective examinations, such as tongue inspection, by trained specialist physicians. An objective metric of the tongue color spectrum was developed as a surrogate for subjective visual inspection. METHODS: Tongue images were acquired with a hyperspectral imaging system, and the uncoated tongue region was segmented automatically. The spectral information of the uncoated tongue area was analyzed by principal component analysis (PCA). The component vector most representative of each clinical symptom was found by rotating the vector on a plane spanned by two arbitrary principal component vectors. RESULTS: The system was tested in human volunteers. Forty-four hyperspectral images were acquired from 30 healthy male subjects for initial testing. The Oketsu (blood stagnation) score was determined by an experienced clinician in Kampo medicine from 27 of 30 subjects. The correlation between respective principal components and Oketsu score was 0.67 at maximum, and increased to 0.73 by linear combination, while it was -0.75 by vector rotation. Significant correlations for many disorders were demonstrated, and vector rotation showed better correlation than linear combination. CONCLUSIONS: A PCA-based algorithm was developed to objectively evaluate patients using color images of the tongue surface. Testing showed that this method was a feasible surrogate for expert visual tongue analysis. This tool should help non-trained people identify "Mibyou" health status for individuals. The algorithm is free of empirical criteria, and it may be it applicable to many hyperspectral image types.

Regional image analysis of the tongue color spectrum.

PURPOSE: Kampo medicine (Japanese traditional herbal medicine) can identify "Mibyou" (disease-oriented state) based on visual tongue inspection by a trained physician. Surface colors of uncoated tongue provide essential features. A hyperspectral imaging system which performs regional image analysis of the tongue color spectrum was developed to automatically inspect the uncoated tongue. METHODS: Hyperspectral tongue surface images were acquired by a camera equipped with an integrating sphere. The color spectrum from 4 tongue areas--uncoated tongue, coated tongue, lip, and perioral areas--were quantified. The average spectrum of each area was determined and 4 images calculated by focusing on the differences between respective spectra; highlight, shadow, tongue coat, and lip-weighted images were calculated. The uncoated tongue area was extracted by subtraction. RESULTS: Reproducibility was evaluated by applying the camera and extraction algorithm to 44 images from human subjects. Eighty-two percentage were judged as acceptable by a Kampo medicine physician expert. CONCLUSIONS: Using a hyperspectral camera and extraction algorithm, the tongue color of the uncoated part was automatically extracted. This technique is suitable for tongue color analysis and may help non-trained users to identify "Mibyou".

Inhibitory Effect of TNF-alpha Produced by Macrophages Stimulated with Grifola frondosa Extract (ME) on the Growth of Influenza A/Aichi/2/68 Virus in MDCK Cells.

We investigated the inhibitory effect of the conditioned medium (CM) from P338D1 (D1) cells, a murine macrophage cell line, stimulated for 10 hours with a fixed dose (100 mug/ml) of the extracts from the fruit bodies of Grifola frondosa (ME) or its ultra filtration-based fractions (MFs), on the growth of influenza A/Aichi/2/68 virus in Madin-Darby canine kidney cells. Direct addition of ME and 3 kinds of MFs (MF1, MF2 and MF3) to the infected cells had no obvious inhibitory effect. However, virus yields were reduced in the presence of CMs. Notably, the inhibitory effect of the CM prepared by using MF2 (molecular weight of 30 Kd to 100 Kd) was the strongest (28% reduction compared to the control). RT-PCR and ELISA assays showed that the CMs could induce the expression of TNF-alpha mRNA in D1 cells leading to production of TNF-alpha, known as an antiviral cytokine. These findings suggest that ME and MFs (especially MF-2) might induce the production of certain factors, including TNF-alpha, which are responsible for the inhibition of viral growth in vitro.

A Proteomic Approach for the Diagnosis of 'Oketsu' (blood stasis), a Pathophysiologic Concept of Japanese Traditional (Kampo) Medicine.

'Oketsu' is a pathophysiologic concept in Japanese traditional (Kampo) medicine, primarily denoting blood stasis/stagnant syndrome. Here we have explored plasma protein biomarkers and/or diagnostic algorithms for 'Oketsu'. Sixteen rheumatoid arthritis (RA) patients were treated with keishibukuryogan (KBG), a representative Kampo medicine for improving 'Oketsu'. Plasma samples were diagnosed as either having an 'Oketsu' (n = 19) or 'non-Oketsu' (n = 29) state according to Terasawa's 'Oketsu' scoring system. Protein profiles were obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and hierarchical clustering and decision tree analyses were performed. KBG treatment for 4 or 12 weeks decreased the 'Oketsu' scores significantly. SELDI protein profiles gave 266 protein peaks, whose expression was significantly different between the 'Oketsu' and 'non-Oketsu' states. Hierarchical clustering gave three major clusters (I, II, III). The majority (68.4%) of 'Oketsu' samples were clustered into one cluster as the principal component of cluster I. The remaining 'Oketsu' profiles constituted a minor component of cluster II and were all derived from patients cured of the 'Oketsu' state at 12 weeks. Construction of the decision tree addressed the possibility of developing a diagnostic algorithm for 'Oketsu'. A reduction in measurement/pre-processing conditions (from 55 to 16) gave a similar outcome in the clustering and decision tree analyses. The present study suggests that the pathophysiologic concept of Kampo medicine 'Oketsu' has a physical basis in terms of the profile of blood proteins. It may be possible to establish a set of objective criteria for diagnosing 'Oketsu' using a combination of proteomic and bioinformatics-based classification methods.

Evidence-based efficacy of Kampo formulas in a model of non alcoholic fatty liver.

Data on the efficacy of herbal compounds are often burdened by the lack of appropriate controls or a limited statistical power. Treatments to prevent the progression of non alcoholic fatty liver disease (NAFLD) to steatohepatitis (NASH) remain unsatisfactory. A total of 56 rabbits were arrayed into 7 groups fed with standard rabbit chow (SRC), SRC with 1% cholesterol, or each of the five experimental treatments (Kampo formulas 1% keishibukuryogan [KBG], 1% orengedokuto [OGT], and 1% shosaikoto [SST]; vitamin E [VE]; or pioglitazone [PG]) in a 1% cholesterol SRC. We analyzed changes after 12 weeks in plasma and liver lipid profiles, glucose metabolism, adipocytokines, oxidative stress, and liver fibrosis. Data demonstrated that all five treatments were associated with significant amelioration of lipid profiles, oxidative stress, and liver fibrosis compared to no supplementation. KBG was superior to VE and PG in the reduction of liver total cholesterol (P < 0.01) and lipid peroxidase levels (P < 0.05), urinary 8-hydroxy-2'-deoxyguanosine (P < 0.05), hepatic alpha-smooth muscle actin positive areas (P < 0.01) and activated stellate cells (P < 0.01). In conclusion, there was a statistically significant benefit of Kampo formulas (KBG in particular) on a dietary model of NAFLD/NASH. Future studies need to be directed at the mechanisms in the treatment of NASH.

Identification of a predictive biomarker for the beneficial effect of a Kampo (Japanese traditional) medicine keishibukuryogan in rheumatoid arthritis patients.

OBJECTIVES: Kampo (Japanese traditional herbal) medicines are now ethically used in Japan as pharmaceutical grade prescription drugs. However, there are distinct groups of responders and non-responders to Kampo medicines. We searched for biomarker candidates to discriminate responders from non-responders to keishibukuryogan (KBG); one of the most frequently used Kampo medicines. DESIGN AND METHODS: A combination of SELDI technology and a decision tree analysis with proprietary developed bioinformatics tools was applied to 41 (32 for tree construction and 9 for validation test) plasma samples obtained from rheumatoid arthritis (RA) patients. A candidate biomarker protein was identified using LC-MS/MS. RESULTS: The constructed tree with measurable reliability contained only a single peak which was identified as haptoglobin alpha 1 chain (Hpalpha1). CONCLUSION: Hpalpha1 is a biomarker candidate for discriminating responders from non-responders to KBG treatment for RA. The present results may open the way to the establishment of "evidence-based" complementary and alternative medicine.

Population of CD40L-expressing cells was slightly but not significantly decreased in lymphoid tissues of collagen-induced arthritic mice treated with Hochu-Ekki-To.

OBJECTIVE: To clarify the mechanism of the action of Hochu-Ekki-To (HET) on collagen-induced arthritic (CIA) mice by analyzing the CD40L-expressing cells population. METHODS: CIA was induced in male DBA/1J mice by immunization with two injections of bovine type II collagen (CII). HET or water was orally administered. The subpopulations of lymphocytes obtained from lymph nodes and spleen were detected at 3 weeks after boost using flow cytometry. RESULTS: Although the population of CD4+CD40L+ cells tended to be decreased in the HET group compared to that in control mice, there was no significant difference between the two groups. These findings were observed in lymphocytes obtained from both lymph nodes and spleen. CONCLUSION: HET suppresses the development of CIA. These effects may be partially induced via the decrease in the population of CD4+CD40L+ cells, but the role of this action is probably limited.

Effects of eppikahangeto, a Kampo formula, and Ephedrae herba against citric acid-induced laryngeal cough in guinea pigs.

To evaluate the efficacy of three common antitussive Kampo formulas, eppikahangeto (EPP), bakumondoto (BAK), and shoseiryutogomakyokansekito (SGM), a new cough model of guinea pig was used, which could specifically induce a laryngeal cough by microinjection of citric acid solution into the larynx. Kampo extract was dissolved in water and the animals were given access ad libitum for 3 days, and then the number of coughs during 10 min was counted. EPP extract decreased the number of coughs dose-dependently (0.3% extract, -22.9 +/- 6.6%, P<0.01; 1.0% extract, -32.4 +/- 5.5%, P<0.01). BAK extract and SGM extract had no significant effect. Intraperitoneal injection of codeine (60 mg/kg) also decreased the number of coughs (-36.1 +/- 9.1%, P<0.05). Furthermore, Ephedrae herba (EH) extract reduced the number of coughs (-18.3 +/- 6.0%, P<0.05), but the extract of EPP without EH did not. These results suggest that EPP has an antitussive effect against laryngeally-induced cough in guinea pigs, and the crucial herbal medicine is EH.

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.

Reduction of perception of chronic fatigue in an observational study of patients receiving 12 weeks of Kampo therapy.

OBJECTIVE: The aim of this study was to observe the influence of Kampo therapy on latent chronic fatigue of patients with chronic diseases. SUBJECTS: One hundred and seventy-three (173) consecutive patients with chronic diseases came to our department for the first time. DESIGN: This was a prospective study. Patients were divided into two groups: a chronic fatigue group (CFG) and a nonchronic fatigue group (NCFG). Based on Kampo diagnosis, both groups were prescribed Kampo formulae as an extract or decoction for 12 weeks. OUTCOME MEASURES: By using questionnaires, patients were assessed concerning their physical and mental types of fatigue, their sleep situation, and their attitude toward work or housekeeping, both before and after 12 weeks of treatment, according to Kampo diagnosis. RESULTS: The mental fatigue, physical fatigue, and sleep scores of both groups, and the work score of CFG, were decreased. The rate of reduction of the fatigue score was significantly greater in CFG than in NCFG. The factor responsible for this difference in fatigue score was physical fatigue. CONCLUSIONS: A reduction of the perception of chronic fatigue was observed in patients receiving 12 weeks of Kampo therapy.

Effect of omega-3 fatty acid-containing phospholipids on blood catecholamine concentrations in healthy volunteers: a randomized, placebo-controlled, double-blind trial.

OBJECTIVE: We previously reported that administration of fish oil rich in docosahexaenoic acid (DHA) increased the plasma ratio of epinephrine to norepinephrine (NE) at rest in young adults who were under chronic stress and that this effect was achieved mainly through depression of NE. However, not many reports have documented the effects of eicosapentaenoic acid (EPA) and DHA on blood catecholamine levels in healthy humans. Therefore, we performed another intervention study to test their effect on catecholamines with healthy subjects under no chronic stress. METHODS: Twenty-one healthy young adults (15 men and 6 women) were randomly assigned to an omega-3 group (n = 9) or a control group (n = 12) in a double-blind manner. Twenty capsules of shellfish-derived lipids containing 762 mg of EPA plus DHA per day were administered to the omega-3 group for 2 mo. The controls took the same amount of placebo capsules. Fasting blood samples after a 30-min rest with a catheter in a forearm vein were obtained at the start and the end of the study for catecholamine measurements. RESULTS: EPA but not DHA concentrations in red blood cells significantly increased in the omega-3 group compared with the control group (P < 0.001). Plasma NE concentrations were significantly decreased in the omega-3 group (from 1.49 +/- 0.39 nmol/L to 1.05 +/- 0.14 nmol/L) compared with the control group (from 1.12 +/- 0.24 nmol/L to 1.39 +/- 0.32 nmol/L) with analysis of covariance (P < 0.001). The differences remained significant (P = 0.01) even after deletion of three subjects in the omega-3 group who had the highest baseline NE values and one in the control group who had the lowest baseline NE value to nullify a significant baseline differences in NE between groups. CONCLUSION: This study demonstrated that EPA plus DHA supplementation lowered plasma NE concentrations in normal volunteers even at the small dose of 762 mg of EPA plus DHA per day. This effect of EPA plus DHA to lower plasma NE concentrations may be important to understand some of the effects of fish oils on diseases.