Temperament in Early Childhood Is Associated With Gut Microbiota Composition and Diversity.

Temperament is a key predictor of human mental health and cognitive and emotional development. Although human fear behavior is reportedly associated with gut microbiome in infancy, infant gut microbiota changes dramatically during the first 5 years, when the diversity and composition of gut microbiome are established. This period is crucial for the development of the prefrontal cortex, which is involved in emotion regulation. Therefore, this study investigated the relationship between temperament and gut microbiota in 284 preschool children aged 3-4 years. Child temperament was assessed by maternal reports of the Children's Behavior Questionnaire. Gut microbiota (alpha/beta diversity and genera abundance) was evaluated using 16S rRNA sequencing of stool samples. A low abundance of anti-inflammatory bacteria (e.g., Faecalibacterium) and a high abundance of pro-inflammatory bacteria (e.g., Eggerthella, Flavonifractor) were associated with higher negative emotionality and stress response (i.e., negative affectivity, ß = -0.17, p = 0.004) and lower positive emotionality and reward-seeking (i.e., surgency/extraversion, ß = 0.15, p = 0.013). Additionally, gut microbiota diversity was associated with speed of response initiation (i.e., impulsivity, a specific aspect of surgency/extraversion, ß = 0.16, p = 0.008). This study provides insight into the biological mechanisms of temperament and takes important steps toward identifying predictive markers of psychological/emotional risk.

Effects of Rikkunshi-To, a Japanese kampo medicine, on donepezil-induced gastrointestinal side effects in mice.

Donepezil, an acetylcholinesterase inhibitor, is associated with gastrointestinal symptoms, such as nausea, vomiting, and anorexia, which may affect adherence to continuous therapy. Since Rikkunshi-To, a Japanese herbal medicine, activates the ghrelin signaling pathway and promotes gastrointestinal function, it is administered to prevent gastrointestinal symptoms. We herein investigated whether donepezil-induced gastrointestinal side effects in mice are ameliorated by Rikkunshi-To and if its therapeutic efficacy is mediated by ghrelin. Since pica behavior, the ingestion of kaolin, correlates with nausea and vomiting in humans, donepezil was intraperitoneally administered with or without Rikkunshi-To daily to mice, and food and kaolin intakes were monitored. The effects of donepezil on intestinal motility and a ghrelin receptor antagonist on donepezil-induced pica behavior, anorexia, and changes in intestinal motility were examined in mice treated with Rikkunshi-To. Pica behavior and anorexia were significantly induced by donepezil and significantly inhibited by Rikkunshi-To. Intestinal motility was significantly suppressed by donepezil and promoted by Rikkunshi-To. Furthermore, the therapeutic effects of Rikkunshi-To were antagonized by the ghrelin receptor antagonist. The present results support the therapeutic efficacy of Rikkunshi-To against donepezil-induced gastrointestinal side effects.

Assessment of the effects of switching oral bisphosphonates to denosumab or daily teriparatide in patients with rheumatoid arthritis.

The aim of this observational, non-randomized study was to clarify the unknown effects of switching oral bisphosphonates (BPs) to denosumab (DMAb) or daily teriparatide (TPTD) in patients with rheumatoid arthritis (RA). The characteristics of the 194 female patients included in the study were 183 postmenopausal, age 65.9 years, lumbar spine (LS) T score -1.8, femoral neck (FN) T score -2.3, dose and rate of taking oral prednisolone (3.6 mg/day) 75.8%, and prior BP treatment duration 40.0 months. The patients were allocated to (1) the BP-continue group (n = 80), (2) the switch-to-DMAb group (n = 74), or (3) the switch-to-TPTD group (n = 40). After 18 months, the increase in bone mineral density (BMD) was significantly greater in the switch-to-DMAb group than in the BP-continue group (LS 5.2 vs 2.3%, P < 0.01; FN 3.8 vs 0.0%, P < 0.01) and in the switch-to-TPTD group than in the BP-continue group (LS 9.0 vs 2.3%, P < 0.001; FN 4.9 vs 0.0%, P < 0.01). Moreover, the switch-to-TPTD group showed a higher LS BMD (P < 0.05) and trabecular bone score (TBS) (2.1 vs -0.7%; P < 0.05) increase than the switch-to-DMAb group. Clinical fracture incidence during this period was 8.8% in the BP-continue group, 4.1% in the switch-to-DMAb group, and 2.5% in the switch-to-TPTD group. Both the switch-to-DMAb group and the switch-to-TPTD group showed significant increases in LS and FN BMD, and the switch-to-TPTD group showed a higher increase in TBS compared to the BP-continue group at 18 months. Switching BPs to DMAb or TPTD in female RA may provide some useful osteoporosis treatment options.

Enhancement of ghrelin-signaling system by Rikkunshi-To attenuates teriparatide-induced pica in rats.

Teriparatide is clinically used for the treatment of osteoporosis; however, nausea is often observed in patients. Its insufficient control affects the ability to continue teriparatide therapy. Rikkunshi-To (RKT), a traditional Japanese herbal medicine, improves the gastrointestinal function via activation of the ghrelin-signaling system. We investigated the therapeutic effects of RKT on teriparatide-induced nausea in rats and the involvement of ghrelin in these effects. We previously reported that ovariectomized rats showed pica (kaolin ingestion), a behavior that can be used to assess nausea in rats, after the subcutaneous administration of teriparatide; thus, the behavior was used as an index of nausea. Ovariectomized rats were fed diets with or without RKT (1%) for 2 weeks, and then they received the subcutaneous injection of teriparatide (400 µg/kg). Teriparatide significantly increased the incidence of pica, while suppressing intestinal motility and plasma ghrelin levels in rats fed normal diets; however, rats fed diets with RKT showed improvements in all of the teriparatide-induced adverse reactions. These therapeutic effects were antagonized by a ghrelin receptor antagonist ([D-Lys3]-GHRP-6; 200 nmol/rat). These findings suggest that the enhancement of ghrelin-signaling is involved in RKT's therapeutic effect, and that RKT is a potentially useful treatment for teriparatide-induced nausea.

[Aging-related frailty and sarcopenia. Kampo medicine for frailty and sarcopenia.]

In Japan, which is entering a super-aged society, the prevention of long-term care and bedridden caused by frailty and sarcopenia have been recognized as an urgent need. The pathology of frailty and sarcopenia includes broad problems of senility. A traditional herbal medicine is known to improve symptoms across the organs, many herbal medicines administered by examining the balance of mind and body are effective for frailty and sarcopenia. "The Safety Drug Therapy Guideline 2015 for the Elderly" was fully revised in 2015. In this guideline, traditional herbal medicines, which are suggested to be useful for the elderly, have been taken up, and the evidence of which is being accumulated. We regarded frailty and sarcopenia as 'kidney-Qi' deficiency, a concept that represents aging phenomenon in Kampo medicine and clarified the anti-sarcopenic effects of Go-sha-jinki-gan, a 'kidney-Qi'-tonifying medicine, using animal experimental models.

Go-sha-jinki-Gan (GJG) ameliorates allodynia in chronic constriction injury-model mice via suppression of TNF-α expression in the spinal cord.

BACKGROUND: Alternative medicine is noted for its clinical effect and minimal invasiveness in the treatment of neuropathic pain. Go-sha-jinki-Gan, a traditional Japanese herbal medicine, has been used for meralgia and numbness in elderly patients. However, the exact mechanism of GJG is unclear. This study aimed to investigate the molecular mechanism of the analgesic effect of GJG in a chronic constriction injury model. RESULTS: GJG significantly reduced allodynia and hyperalgesia from the early phase (von Frey test, p<0.0001; cold-plate test, p<0.0001; hot-plate test p»0.011; two-way repeated measures ANOVA). Immunohistochemistry and Western blot analysis revealed that GJG decreased the expression of Iba1 and tumor necrosis factor-a in the spinal cord. Double staining immunohistochemistry showed that most of the tumor necrosis factor-a was co-expressed in Iba1-positive cells at day 3 post-operation. GJG decreased the phosphorylation of p38 in the ipsilateral dorsal horn. Moreover, intrathecal injection of tumor necrosis factor-a opposed the anti-allodynic effect of GJG in the cold-plate test. CONCLUSIONS: Our data suggest that GJG ameliorates allodynia in chronic constriction injury model mice via suppression of tumor necrosis factor-a expression derived from activated microglia. GJG is a promising drug for the treatment of neuropathic pain induced by neuro-inflammation.

Effects of a traditional herbal medicine on peripheral blood flow in women experiencing peripheral coldness: a randomized controlled trial.

BACKGROUND: In Japan, a traditional herbal medicine, Tokishigyakukagoshuyushokyoto (TJ-38), is often used for the treatment of peripheral coldness, which is a common complaint among Japanese women. However, the effects of this herbal medicine have yet to be examined in a randomized controlled trial. In the current study, the effect of TJ-38 on the peripheral blood flow in women experiencing peripheral coldness was investigated using a parallel-group randomized controlled trial. METHODS: Fifty-eight women aged 23 to 79 years with peripheral coldness were randomly divided into the intervention or control group. They were examined using cold bathing tests, physical examinations, and questionnaires in January 2010 for the baseline and in March 2010 for the follow-up, and January 2011 and March 2011, respectively. RESULTS: At the baseline, there were no differences in clinical characteristics between the two groups. In the intervention group, peripheral coldness improved after the intervention term; however, it persisted in the control group. Mean values of percentage recovery of the peripheral blood flow after cold bathing tests were 17.2% and -28.2% for the intervention and control groups, respectively (p = 0.007), and the proportions for percentage recovery of >50% were 32% and 0%, respectively (p = 0.0007). Mean values of percent recovery of skin temperature did not differ between the two groups. CONCLUSIONS: The present clinical trial supports that a traditional herbal medicine relieves peripheral coldness in women probably through the improvement of peripheral blood flow.

Expansion of range of joint motion following treatment of systemic sclerosis with tocilizumab.

Systemic sclerosis (SSc) presents stiffness of extremities due to sclerosis of the tissue especially at fingers, hands, and forearms. Here we report the case of a patient with diffuse cutaneous SSc who was administered anti-interleukin-6 receptor antibody tocilizumab (TCZ). Skin condition of SSc is evaluated by pinching the skin according to the Rodnan skin score, but sometimes tissue atrophy results in overestimation of the condition. To understand how the extremities softened after initiation of TCZ, we observed mobility of extremities. Range of motion (ROM) of joints was measured every four months after initiation of TCZ. The patient presented not only reduction of Rodnan score but also amelioration of mobility of extremities. The Rodnan skin score reduced from 35 to 7 within sixteen months, and ROM of most joints except ankle was expanded.

Amelioration of small bowel injury by switching from nonselective nonsteroidal anti-inflammatory drugs to celecoxib in rheumatoid arthritis patients: a pilot study.

BACKGROUND/AIMS: Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used in patients with rheumatoid arthritis (RA) but have several side effects including mucosal damage in the small intestine. We aimed to evaluate whether the small bowel injury is ameliorated by switching from nonselective NSAIDs to celecoxib in patients with RA. METHODS: Sixteen patients with RA who were treated with nonselective NSAIDs were enrolled in this study. Nonselective NSAIDs were converted to celecoxib for 12 weeks. Capsule endoscopy was performed before and after treatment with celecoxib. Videos were screened by gastroenterologists blinded to the patients' treatment. RESULTS: Before the administration of celecoxib, reddened folds, denuded areas, petechiae/red spots and mucosal breaks were observed in 63, 63, 88 and 69% of the patients, respectively. In the 14 patients who completed this study, conversion to celecoxib significantly reduced the number of petechiae/red spots, the number of mucosal breaks, and Lewis scores. RA activity and cytokine levels in the peripheral blood were not significantly different before and after treatment with celecoxib. CONCLUSIONS: The incidence of small bowel injury by nonselective NSAIDs is high in patients with RA. Conversion from nonselective NSAIDs to celecoxib can be useful for protecting patients with RA from small bowel injury.

Intestinal microbiome and maternal mental health: preventing parental stress and enhancing resilience in mothers.

The number of mothers suffering from mental illness is increasing steadily, particularly under conditions of the coronavirus pandemic. The identification of factors that contribute to resilience in mothers is urgently needed to decrease the risks of poor physical and psychological health. We focused on the risk of parenting stress and psychological resilience in healthy mothers with no psychiatric and physical disorders and conducted two studies to examine the relationships between intestinal microbiota, physical condition, and psychological state. Our results showed that alpha diversity and beta diversity of the microbiome are related to high parenting stress risk. Psychological resilience and physical conditions were associated with relative abundances of the genera Blautia, Clostridium, and Eggerthella. This study helps further understand the gut-brain axis mechanisms and supports proposals for enhancing resilience in mothers.

Altered Gut Microbiota Composition Is Associated with Difficulty in Explicit Emotion Regulation in Young Children.

Executive function (EF) consists of explicit emotion regulation (EER) and cognitive control (CC). Childhood EER in particular predicts mental and physical health in adulthood. Identifying factors affecting EER development has implications for lifelong physical and mental health. Gut microbiota (GM) has attracted attention as a potential biomarker for risk of physical and mental problems in adulthood. Furthermore, GM is related to brain function/structure, which plays a crucial role in emotional processing. However, little is known about how GM compositions are associated with the development of emotion regulation in early childhood. Therefore, in this study, we examined 257 children aged 3-4 to investigate links between GM and risk to EF. EF was measured using the Mother-Reported Behavior Rating Inventory of Executive Function-Preschool version. GM composition (alpha/beta diversity and genus abundance) was evaluated using 16S rRNA gene sequencing and compared between EF-risk and non-risk groups. Our results show that children with EER-risk (an index of inhibitory self-control) had a higher abundance of the genera Actinomyces and Sutterella. Although we have not established a direct link between GM and CC risk, our findings indicate that GM of preschoolers is closely associated with emotional processing and that EERrisk children have more inflammation-related bacteria.

Long-Term Effects of a Ketogenic Diet for Cancer.

A ketogenic diet has been proposed as a potential supportive therapy for cancer patients, although its long-term influence on survival rates remain controversial. In our previous report, we presented promising results for 37 of 55 patients with advanced cancer enrolled between 2013 and 2018 who remained on a ketogenic diet for at least 3 months. We followed all 55 patients until March 2023 and analyzed the data up to March 2022. For the 37 patients with previously reported promising results, the median follow-up period was 25 (range of 3-104) months and 28 patients died. The median overall survival (OS) in this subset of 37 patients was 25.1 months and the 5-year survival rate was 23.9%. We also evaluated the association between the duration of the ketogenic diet and outcome in all 55 patients, except for 2 patients with insufficient data. The patients were divided into two groups: those who followed the diet for ≥12 months (n = 21) and those who followed it for <12 months (n = 32). The median duration of the ketogenic diet was 37 (range of 12-99) months for the ≥12 months group and 3 (range of 0-11) months for the <12 months group. During the follow-up period, 41 patients died (10/21 in the ≥12 months group and 31/32 in the <12 months group). The median OS was 19.9 months (55.1 months in the ≥12 months group and 12 months in the <12 months group). Following the inverse probability of treatment weighting to align the background factors of the two groups and make them comparable, the adjusted log-rank test showed a significantly better OS rate in the group that continued the ketogenic diet for a longer period (p < 0.001, adjusted log-rank test). These results indicate that a longer continuation of the ketogenic diet improved the prognosis of advanced cancer patients.

Ketogenic effects of medium chain triglycerides containing formula and its correlation to breath acetone in healthy volunteers: a randomized, double-blinded, placebo-controlled, single dose-response study.

The efficacy of low-carbohydrate, high-fat diets, such as ketogenic diets, for cancer patients is of research interest. We previously demonstrated the efficacy of the ketogenic diet in a case study in which medium-chain triglycerides (MCTs) or MCT-containing formula (ketogenic formula) was used as a supplement to increase blood ketone bodies. However, little is known about the amounts needed to induce ketogenic effects and about the usefulness of monitoring of breath acetone. To investigate the pharmacokinetics of MCTs and their metabolites, blood ketone bodies and breath acetone, 24 healthy subjects received one of four single oral doses of the ketogenic formula (equivalent to 0, 10, 20, and 30 g of MCTs) under fasting conditions. Total blood ketone bodies, ß-hydroxybutyrate, octanoic acid, and decanoic acid were increased in a dose-dependent manner. The ketogenic effect was considered to depend on octanoic and decanoic acids, because a positive correlation was observed between them. A strong positive correlation was also observed between total serum ketone bodies and breath acetone at each time points. Therefore, monitoring breath acetone levels seems a less invasive method to predict blood concentrations of ketone bodies during ketogenic diet therapy. Clinical trial registration:https://rctportal.niph.go.jp/en/detail?trial_id=UMIN000032634, UMIN-CTR UMIN000032634.

A case of Behçet's disease treated with a humanized anti-interleukin-6 receptor antibody, tocilizumab.

A 47-year-old female patient with Behçet's disease had been treated with colchicine, prednisolone, cyclosporine A, and infliximab. Because she relapsed, however, treatment with tocilizumab, a humanized anti-interleukin 6 receptor antibody, was started. This treatment suppressed the patient's clinical manifestations, including ocular attacks, for 1 year and improved her visual acuity. This experience indicates that tocilizumab may constitute a therapeutic option for refractory Behçet's disease.

Gosha-jinki-Gan (GJG) shows anti-aging effects through suppression of TNF-α production by Chikusetsusaponin V.

Frailty develops due to multiple factors, such as sarcopenia, chronic pain, and dementia. Go-sha-jinki-Gan (GJG) is a traditional Japanese herbal medicine used for age-related symptoms. We have reported that GJG improved sarcopenia, chronic pain, and central nervous system function through suppression of tumor necrosis factor-alpha (TNF-α) production. In the present study, GJG was found to reduce the production of TNF-α in the soleus muscle of senescence-accelerated mice at 12 weeks and 36 weeks. GJG did not change the differentiation of C2C12 cells with 2% horse serum. GJG significantly decreased the expression of Muscle atrophy F-box protein (MAFbx) induced by TNF-α in C2C12 cells on real-time PCR. TNF-α significantly decreased the expression of PGC-1α and negated the enhancing effect of GJG for the expression of PGC-1α on digital PCR. Examining 20 chemical compounds derived from GJG, cinnamaldehyde from cinnamon bark and Chikusetsusaponin V (CsV) from Achyrantes Root dose-dependently decreased the production of TNF-⍺ in RAW264.7 cells stimulated by LPS. CsV inhibited the nuclear translocation of nuclear factor-kappa B (NF-κB) p65 in RAW264.7 cells. CsV showed low permeability using Caco-2 cells. However, the plasma concentration of CsV was detected from 30 min to 6 h and peaked at 1 h in the CD1 (ICR) mice after a single dose of GJG. In 8-week-old SAMP8 mice fed 4% (w/w) GJG from one week to four weeks, the plasma CsV concentration ranged from 0.0500 to 10.0 ng/mL. The evidence that CsV plays an important role in various anti-aging effects of GJG via suppression of TNF-⍺ expression is presented.

The Japan Frailty Scale is a promising screening test for frailty and pre-frailty in Japanese elderly people.

Frailty is one of the most important problems in a super-aged society. It is necessary to identify frailty quickly and easily at the bedside. We developed a simple patient-reported frailty screening scale, the Japan Frailty Scale (JFS), based on the aging concept of Kampo medicine. Eight candidate questions were prepared by Kampo medicine experts, and a simple prediction model was created in the development cohort (n = 434) and externally validated in an independent validation cohort (n = 276). The physical indicators and questionnaires associated with frailty were also comprehensively evaluated. The reference standard for frailty or pre-frailty was determined based on the Kihon checklist. In the development cohort, four questions, nocturia (0-2), lumbago (0-2), cold sensitivity (0-2), exhaustion (0-4), and age (0-1) were selected by multivariable logistic regression analysis. The total JFS score is 0-11. Receiver-operating characteristic curve analysis of the JFS for identifying frailty status showed moderately good discrimination (area under the curve (AUC) = 0.78, 95 % confidence interval (CI): 0.73-0.82). At the JFS cutoff value of 3/4 for frailty or pre-frailty, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 86.9 %, 53.3 %, 62.8 %, and 81.7 %, respectively. External validation of the JFS showed moderately good discrimination (AUC = 0.76, 95 % CI: 0.70-0.81). The sensitivity, specificity, PPV, and NPV were 79.9 %, 61.4 %, 69.3 %, and 73.7 %, respectively. These results indicate that the JFS is a promising patient-reported clinical scale for early identification of pre-frail/frail patients at the bedside in primary care.

Clinical Importance of Body Composition in Improving Bone Mineral Density of Femoral Neck After Denosumab Therapy in Patients With Rheumatoid Arthritis or Collagen Diseases.

BACKGROUND/AIM: To investigate factors associated with increased bone mineral density (BMD) of the neck of femur in rheumatoid arthritis or collagen diseases receiving denosumab, focusing on body composition calculated by bioelectrical impedance analysis (n=90, 78 females). PATIENTS AND METHODS: We defined Δfemur as BMD (12 months minus baseline), using dual-energy X-ray absorptiometry after denosumab therapy. Factors associated with Δfemur were retrospectively investigated. RESULTS: Low skeletal muscle index (SMI) was observed in 6 males and 32 females. There was a significant difference in phase angle (PhA) of the left leg (LL) between the Δfemur ≥0 (n=70) and Δfemur <0 (n=20) groups (p=0.040) but not in SMI (p=0.310). Multiple regression analysis indicated that PhA of LL was significantly related to Δfemur (p=0.0398). CONCLUSION: PhA appears to be a clinically significant indicator of improvement of Δfemur in patients receiving denosumab.

A multidimensional physical scale is a useful screening test for mild depression associated with childcare in Japanese child-rearing women.

Introduction: Maternal depression is one of the important problems of postpartum women. For its early detection and appropriate treatment, it is necessary to identify women at high risk for depression quickly and easily. Materials and methods: A simple screening scale for depression from physical aspects, the multidimensional physical scale (MDPS), which is a 17-item, self-report, three-step scale (0, 1, 2) according to the theory of Kampo medicine, was developed. The aim of the present study was to develop (n = 785) and validate (n = 350) the MDPS that was designed to rate the risk of depression. The Beck Depression Inventory-Second Edition was used for determination of depression. In the development cohort, the final model was determined using multi-regression logistic analysis. Results: The components of the MDPS for mothers (MDPS-M) were developed, containing the total score of MDPS (0-34 points) and resumption of menstruation or not (-3, 0 points). Receiver-operating characteristic curve analysis of the MDPS-M (-3 to 34) for identifying a high risk of depression showed moderately good discrimination [area under the curve (AUC) = 0.74, 95% confidence interval (CI): 0.70-0.78]. At the cutoff value of MDPS-M (9/10), its sensitivity, specificity, positive predictive value, and negative predictive value were 84.9, 45.7, 36.7, and 89.2%, respectively. External validation of the MDPS-M showed moderately good discrimination (AUC = 0.74, 95% CI: 0.68-0.79) using the same analysis as the development cohort. Conclusion: These results indicate that the MDPS-M is a useful, simple, clinical scale for early identification of mothers at high risk of depression in primary care.

Ketogenic Effects of Multiple Doses of a Medium Chain Triglycerides Enriched Ketogenic Formula in Healthy Men under the Ketogenic Diet: A Randomized, Double-Blinded, Placebo-Controlled Study.

Ketogenic diets, which are carbohydrate-restricted high-fat diets, may have therapeutic effects on various diseases, including cancer. However, ketogenic diets are often not standardized and, therefore, results are difficult to interpret. We previously investigated the usefulness of ketogenic diets in cancer therapy, where ketogenic formulas (KF) were used as supplements to enhance blood ketone bodies; however, the amount of KF was determined empirically with reference to blood ketone bodies levels. Here, to determine a standardized optimal amount of KF, we investigated temporal changes in blood ketone bodies (acetoacetic acid (AcAc), ß-hydroxybutyrate (BHB)) and safety in 20 healthy individuals when KF was taken repeatedly under the conditions of a ketogenic diet (UMIN000034216). The diurnal variation in total ketone bodies, and AcAc and BHB levels significantly increased after lunch and after dinner, on the 4th day of KF administration. There were no significant safety issues related to KF in the context of anthropometric, metabolic, nutritional, urological and gastrointestinal parameters. In addition, ketogenic diets lead to changes in gut microbiota. KF showed a decrease in phylum Firmicutes. Our study provides baseline data of the usefulness of KF in a ketogenic diet.

Suppressive effects of processed aconite root on dexamethasone-induced muscle ring finger protein-1 expression and its active ingredients.

Processed aconite root (PA), the tuberous root of Aconitum carmichaelii prepared by autoclaving, is a crude drug used in Japanese traditional Kampo medicine and traditional Chinese medicine for the symptoms of kidney deficiency, that is related to the muscle atrophy in modern medicine. The objective of the present study is to evaluate the effectiveness of PA on muscle atrophy and to find its active ingredients using dexamethasone-induced muscle ring finger protein-1 (MuRF1) mRNA expression in murine myoblast C2C12 cells. Dexamethasone-induced MuRF1 expression was significantly suppressed by methanol-soluble part of boiling water extract of PA in a concentration-dependent manner with its IC50 value of 1.5 mg/ml. By the activity-guided fractionations of PA extract using the partition between organic solvents and its aqueous solution, the activity of PA did not transfer into the fraction containing aconitine-type diterpenoid alkaloids but into BuOH layer. Then, we found higenamine and salsolinol as the active ingredients in PA. Higenamine and salsolinol significantly suppressed dexamethasone-induced MuRF1 expression, and their IC50 values were 0.49 and 50 µM, respectively. The contents of higenamine and salsolinol in the decoctions of commercially available fourteen PA products are 0.12 and 14 µg/ml as the average values, and varied with the coefficient of variation (CV) values of 97 and 63%, respectively. Higenamine also significantly suppressed dexamethasone-induced mRNA expressions of muscle atrophy F-box protein (MAFbx)/atrogin1, casitas B-lineage lymphoma-b (Cbl-b), troponin, branched-chain amino acid aminotransferase 2 (BCAT2), and Bcl-2 binding and pro-apoptotic protein3 (Bnip3). Although the quality control of PA is regulated by the contents of diterpene alkaloids, salsolinol and higenamine can be used as the marker compounds to certificate the pharmacological activities of PA.