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The detection method of characteristic spectrum for reference samples was established by preparing 15 batches of the reference samples of Huagan Decoction, and the peak attribution and the similarity range in the characteristic spectrum were clarified. The ranges of paste-forming rate, content, and transfer rate of the index components including geniposide, paeonol, and paeoniflorin were analyzed. The key quality attribute of the reference samples of Huagan Decoction was defined. The results showed that the 15 batches of the reference samples of Huagan Decoction had good similarities in the characteristic spectrum, which were all higher than 0.9. According to the information of characteristic peak, there were 18 characteristic peaks in the whole prescription, including seven common characteristic peaks from green tangerine peel and dried tangerine peel, four characteristic peaks from tree peony root bark(three of them were common characteristic peaks from tree peony root bark and red peony root), five characteristic peaks from cape jasmine fruit, one characteristic peak from paniculate bolbostemma, and one characteristic peak from oriental waterplantain rhizome.The paste-forming rate of the 15 batches of reference samples was 14.73%-18.83%. The content of geniposide was 1.68%-2.87%, with the average transfer rate of 70.05%±11.13%. The content of paeonol was 0.10%-0.16%, with the average transfer rate of 9.38%±1.78%. The content of paeoniflorin was 1.94%-2.74%, with the average transfer rate of 36.69%±4.63%. This study analyzed the quality value transfer of the reference samples of Huagan Decoction by the evaluation mode of combining the characteristic spectrum, the paste-forming rate, and the content of index components. The findings of this study initially established a stable and feasible standard decoction evaluation method and provided references for the quality control and the subsequent development of relevant preparations of Huagan Decoction.

Objective: Asthma and allergic rhinitis are chronic inflammatory diseases of the conducting nasal airway. Traditional Chinese medicine has long been used for supplemental therapy of allergic diseases, especially asthma and allergic rhinitis. We previously reported the effects of a mixed herbal formula in patients with allergic rhinitis. However, the immune-modulatory mechanism underlying the effects of herbal medicine for the treatment of allergic diseases remains unclear., Methods: We investigated the physiologic changes in dendritic cell (DC) and CD4(+) T cell activity in patients with asthma and allergic rhinitis who were treated with a mixed herbal formula composed of Xin-yi-san + Xiao-qing-long-tang + Xiang-sha-liu- jun-zi-tang. Specifically, we set up in vitro autologous or heterologous co-culture experiments between DCs and CD4(+) T cells, and used flow cytometry and ELISA to analyze the expression of surface molecules on DCs and the release of cytokines by CD4(+) T cells., Results: Expression of HLA-DR on DCs was suppressed following treatment with the mixed herbal formula. Surface expression of CD40, CD54 and CD86 on DCs was also attenuated after treatment. In autologous co-cultures, CD4(+) T cells increased their IL-10 production while decreasing TNF-α production. In heterologous co-cultures, IL-10 secretion by T cells was enhanced, while IL-12, IL-4, IL-5 and TNF-α secretion were reduced., Conclusion: Our mixed herbal formula attenuated the allergic reaction by modifying the physiologic function of the DC-CD4(+) T cell interaction. Further investigations are necessary to understand the mechanism of immune modification mediated by the mixed herbal formula.

Aim: Cyclovirobuxinum D (CVB-D), an alkaloid isolated from the Chinese medicinal plant Buxus microphylla, has been found to be effective to treat cardiac insufficiency, arrhythmias and coronary heart disease. In the present study, we investigated the effects of CVB-D on the inflammatory responses in lipopolysaccharide (LPS)-stimulated murine macrophages in vitro and the underlying mechanisms., Methods: Murine macrophage cell line RAW264.7 cells were incubated in the presence of LPS (0.1 μg/mL) for 24 h. The cell viability was measured using MTT assay. The release of NO and cytokines were detected using the Griess test and ELISA, respectively. The mRNA and protein levels were determined using RT-PCR and Western blot, respectively. Reporter gene assays were used to analyze the transcriptional activity of NF-κB., Results: Treatment of RAW264.7 cells with CVB-D (25-300 μmol/L) did not affect the cell viability. Pretreatment with CVB-D (50, 100 and 200 μmol/L) concentration-dependently decreased NO release and iNOS expression in LPS-treated RAW264.7 cells (its IC50 value in inhibition of NO production was 144 μmol/L). CVB-D also concentration-dependently inhibited the secretion and mRNA expression of IL-1β and IL-6 in LPS-treated RAW264.7 cells. Furthermore, CVB-D remarkably inhibited the phosphorylation of STAT1 and STAT3, as well as JAK2 in LPS-treated RAW264.7 cells, but did not affect the activation of NF-κB and MAPKs pathways. Pretreatment with the JAK2 specific inhibitor AG490 (30 μmol/L) produced similar effects on NO release and iNOS expression in LPS-treated RAW264.7 cells., Conclusion: CVB-D exerts anti-inflammatory effects in LPS-stimulated murine macrophages in vitro at least in part by blocking the JAK-STAT signaling pathway. The anti-inflammatory actions of CVB-D may contribute to its cardioprotection.

Three new compounds, 3,6-dihydroxy-4,5-dimethoxy-1,8-naphalic anhydride (1), 3,4,5,6-tetrahydroxy-1,8-naphalic anhydride (2), and methyl (7E,9E)-6,11-dioxononadeca-7,9-dienoate (3), were isolated from the stem bark of Juglans mandshurica. Their structures were elucidated on the basis of spectroscopic evidence, including 1D and 2D NMR, HR-TOF-MS, and by comparison with the literature data.

Background: Drug interactions between traditional Chinese herbal medicines and the anticoagulant warfarin may cause patient harm and are, therefore, important in clinical practice. Our experience in daily practice suggests that prothrombin time (PT) is prolonged when warfarin is used in combination with the Chinese herbal formula Shu-Jing-Hwo-Shiee-Tang (SJHST) commonly used by patients with osteoarthritis., Objective: We conducted animal experiments to confirm the effect of SJHST and warfarin on anticoagulant activity., Methods: Forty-eight male New Zealand white rabbits were randomized into eight groups of six rabbits. Group A (Control group) was administered normal saline. Group B (Western Medicine group) was administered warfarin 1.5 mg/kg/day. Groups C, D, and E [Traditional Chinese Medicine (TCM) groups] were administered different doses of SJHST (0.5 mg/kg/day, 1 mg/kg/day, and 2 mg/kg/day, respectively). Groups F, G, and H (Combination Therapy groups) were administered warfarin 1.5 mg/kg/day and different doses of SJHST (0.5 mg/kg/day, 1 mg/kg/day, and 2 mg/kg/day, respectively). The total duration of treatment was 14 days. Blood samples were obtained prior to beginning the experiments (day 0) and on day 7, day 14, and day 17 (3 days after discontinuation of the medications). The activated partial thromboplastin time (APTT), PT, and thrombin time (TT) were calculated and compared among the different groups., Results: No significant changes were noted in APTT, PT or TT between the control and SJHST-only groups. Significant prolongations of APTT and PTT, but not TT, were observed in the combination groups compared to the warfarin-only group. The enhanced anticoagulant effects returned to normal three days after discontinuation of SJHST treatment., Conclusions: We confirmed that the Chinese herb SJHST enhances the anticoagulant effect of warfarin. Although the exact mechanisms of the interaction are unknown, physicians should be aware of the possibility of drug interactions between warfarin and Chinese herbal medicines owing to the increased risk of bleeding.

Objective: To compare Cirsium japonicum characteristics with C. leo and C. leducei, along with the content of buddleoside and pectolinarin, and lay the foundation for the quality control of C. japonicum., Method: Samples were collected and the relevant drugs were bought. The samples were divided into root, stem, leaf and flower, and the content of buddleoside and pectolinarin was determine by the HPLC. Chromatographic column: Waters XBridge C18 (4.6 mm x 250 mm), mobile phase: methanol-water (45: 55), measurement wavelength: 326 nm, flow rate: 0.8 mL x min(-1), column temperature: 30 degrees C. RESULT AND CONDUSION: Standard curve equation of buddleoside: Y = 74 064X-47 748, R2 = 0.991. Standard curve equation of pectolinarin: Y = 1 711 64X - 180 707, R2 = 0.999. The content of buddleoside: C. japonicum leaf was 1.987 3%, C. leo leaf 1.412 2%, C. leducei leaf 0.149 2%. The content of buddleoside was lower in root and stem. Pectolinarin was not detected in the C. japonicum and C. leo. The pectolinarin content was 0.069 0% in C. leducei leaf.

A new 2,5-diketopiperazine, (R)-2-(2-(furan-2-yl)-oxoethyl)-octahydropyrrolo[1,2-a]pyrazine-1,4-dione, and seven known compounds were isolated from the ethyl acetate extract of liquid fermentation broth of Armillaria mellea. The structures of the isolated compounds were established from NMR and HR-MS data. The absolute configuration of the new compound was established by comparing the experimental electronic circular dichroism (ECD) spectrum with the calculated ECD data.

Juglanthraquinone C (1,5-dihydroxy-9,10-anthraquinone-3-carboxylic acid, JC), a naturally occurring anthraquinone isolated from the stem bark of Juglans mandshurica, shows strong cytotoxicity in various human cancer cells in vitro. Here, we first performed a structure-activity relationship study of six anthraquinone compounds (JC, rhein, emodin, aloe-emodin, physcion and chrysophanol) to exploit the relationship between their structural features and activity. The results showed that JC exhibited the strongest cytotoxicity of all compounds evaluated. Next, we used JC to treat several human cancer cell lines and found that JC showed an inhibitory effect on cell viability in dose-dependent (2.5-10 μg/ml JC) and time-dependent (24-48 h) manners. Importantly, the inhibitory effect of JC on HepG2 (human hepatocellular carcinoma) cells was more significant as shown by an IC(50) value of 9 ± 1.4 μg/ml, and 36 ± 1.2 μg/ml in L02 (human normal liver) cells. Further study suggested that JC-induced inhibition HepG2 cell proliferation was associated with S phase arrest, decreased protein expression of proliferation marker Ki67, cyclin A and cyclin-dependent kinase (CDK) 2, and increased expression of cyclin E and CDK inhibitory protein Cip1/p21. In addition, JC significantly triggered apoptosis in HepG2 cells, which was characterized by increased chromatin condensation and DNA fragmentation, activation of caspase-9 and -3, and induction of a higher Bax/Bcl2 ratio. Collectively, our study demonstrated that JC can efficiently inhibit proliferation and induce apoptosis in HepG2 cells.

Objective: To observe effects of Dan Wei Powder (Powder for treating the gall bladder and stomach) Tea Bag (DWSTB) on the aggregation rate of blood platelet in vivo and in vitro., Methods: Increase of the platelet aggregation rate in the rat in vivo was induced by carrageenin, and increase of the rabbit platelet aggregation rate in vitro was induced by adenosine diphosphate (ADP) and collagen, respectively. The effects of DWSTB on the platelet aggregation rate were investigated in vivo and in vitro, respectively., Results: The maximum in vivo platelet aggregation rate in the rat was significantly decreased after administration of 2.0 and 4.0 g x kg(-1) DWSTB (P < 0.05, P < 0.01). The maximum rabbit platelet aggregation rate induced by ADP and collagen in vitro were suppressed significantly by 2.0-16.0 mg x mL(-1) and 2.0-8.0 mg x mL(-1) DWSTB, respectively (P < 0.05, P < 0.01). And the effect of DWSTB on platelet aggregation was raised with increase of its dose., Conclusion: Dan Wei Powder Tea Bag can restrain the aggregation of platelet in vivo and in vitro.

Aims of Study: Ligusticum wallichi Franchat (chuanxiong) is a very common traditional Chinese herbal medicine in China. Tetramethylpyrazine (TMP) is a major active ingredient extracted from Ligusticum wallichi Franchat. We investigated the protective effect of TMP on interleukin-1β (IL-1β) induced proteoglycan (PG) degradation and apoptosis in rabbit articular cartilage and chondrocytes., Materials and Methods: Rabbit articular cartilage explants and chondrocytes were cultured with 10 ng/ml IL-1β for 72 h in the absence or presence of various concentrations of TMP (50, 100 or 200 μM). Cartilage and chondroprotective effects of TMP were determined by evaluating (1) the degree of PG degradation by measuring the amount of glycosaminoglycan (GAG) released into the culture media with 1,9-dimethylmethylene blue (DMMB) assay in cartilage explants; (2) gene expression of MMP-3 and TIMP-1 by real-time quantitative reverse transcription-polymerase chain reaction analysis in cartilage explants; (3) chondrocytes viability with MTT assay; (4) the production of intracellular reactive oxygen species (ROS) with laser scanning confocal microscopy (LSCM). Anti-apoptotic effects of TMP were determined by measuring (1) apoptosis with flow cytometric analysis; (2) mitochondrial membrane potential assay with LSCM; (3) caspase-3 activity with special assay kit., Results: IL-1β treatment increased the level of GAG released into the culture media, and induced the gene expression of MMP-3 and inhibited the gene expression of TIMP-1 in cartilage explants. Moreover, IL-1β treatment decreased the cell viability and mitochondrial membrane potential, and enhanced the level of intracellular ROS, apoptosis rate, and caspase-3 activity in chondrocytes. However, simultaneous treatment with TMP attenuated the IL-1β-induced cartilage and chondrocyte destruction in a dose-dependent manner. TMP showed the decrease of GAG degradation and MMP-3 mRNA production, and the enhancement of TIMP-1 mRNA production in cartilage explants. TMP also increased the cell viability in chondrocytes. Furthermore, TMP inhibited the chondrocytes apoptosis through suppression of ROS production, maintaining of mitochondrial membrane potential and downregulation of caspase-3 activity., Conclusion: These results demonstrate that TMP has the cartilage and chondroprotective effect, which suggest that TMP could act as an agent for pharmacological intervention in the progress of OA., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Background: Perennial allergic rhinitis (AR) is a common health problem with a high prevalence rate of 43.6% in Taiwan. In our previous study, a mixed formula of Chinese herbs consisted of Xin-yi-san, Xiao-ging-long-tang, and Xiang-sha-liu-jun-zi-tang, exerted diverse immunomodulatory effects in the treatment of patients with perennial allergic rhinitis., Objective: The aim of the present study was to determine whether Xin-yi-san (XYS) alone is effective in the treatment of AR and tried to explore its molecular mechanism of anti-allergic activity., Methods: In a randomized double-blind study, 108 patients with AR received either XYS or a placebo for 3 months. The effectiveness of XYS was evaluated by nasal symptoms, nasal airflow resistance, nostril dissection area, and serum titer of specific IgE antibodies against house dust mite allergens. In addition, the production of T helper (Th) 1 (represented by interferon-gamma) and Th2 [represented by interleukin (IL)-4, IL-10, and IL-13] cytokines, the proinflammatory cytokine IL-8, soluble intercellular adhesion molecule (sICAM), and arachidonate metabolites prostagladin E(2) (PGE(2)) and leukotriene C(4) (LTC(4)) by polymorphonuclear neutrophils (PMNs) were compared before and after treatment between the two groups., Results: XYS attenuated nasal symptoms (sneezing and rhinorrhea) and nasal congestion through reduction of nasal airflow resistance and increase in nostril dissection areas. We also found that XYS exerted diverse immunomodulatory effects, including suppression of serum IgE levels and increased production of IL-10, sICAM-1, and IL-8 compared to placebo group. However, XYS treatment did not affect the release of PGE(2) and LTC(4) from PMNs., Conclusions: This study originally provides the evidence that Xin-yi-san alone is an effective herb in the treatment of perennial allergic rhinitis., ((c) 2010 Elsevier B.V. All rights reserved.)

Bu-zhong-yi-qi-tang, an ancient formula of Chinese medicine usually used in the treatment of allergic diseases, was evaluated in the treatment of patients with perennial allergic rhinitis. In this study, 60 patients allergic to house dust mite allergen confirmed by skin test and MAST test were recruited and randomized. An experimental group of 36 patients was treated with Bu-zhong-yi-qi-tang, whereas a control group of 24 patients was treated with a non-effective formula Ping-wei-san for 3 months. The nasal symptomatic scores and the responses of polymorphonuclear neutrophils (PMN) to IL-4-stimulation were measured after treatment. The nasal symptomatic scores in the experimental group were significantly improved (3.78+/-0.09 before treatment vs. 0.57+/-0.06 after treatment). In contrast, no change was found in symptomatic scores in the control group (3.17+/-0.12 before treatment vs. 2.79+/-0.14 after treatment). Moreover, total serum IgE and the IL-4-stimulated production of PGE(2) and LTC(4) by PMN was significantly suppressed in the experimental group after treatment compared to the control group. The COX-2 mRNA expression in IL-4-stimulated PMN was also significantly suppressed after Bu-zhong-yi-qi-tang treatment. These results suggest that Bu-zhong-yi-qi-tang but not Ping-wei-san was beneficial to the patients with perennial allergic rhinitis via suppressed nasal inflammation by an antiinflammatory effect.

Objective: To observe the effects of refined Xuefu Capsule (RXC) in patients with arteriosclerosis obliterans (ASO) after femoral-popliteal bypass (FPB)., Methods: Thirty-eight ASO patients with 41 limbs underwent FPB were randomly assigned to two groups: the control group (18 cases with 20 operated limbs) and the RXC group (20 cases with 21 operated limbs). All patients received long-term anticoagulant treatment with Warfarin, and RXC was given to the RXC group additionally for 3 months. Changes of clinical symptoms, blood coagulating function, as well as the condition of vascular patency and ankle arm index (AAI) were assessed 6 and 12 months after operation., Results: Clinical symptoms were improved in all the patients after operation. One year after operation, incidence of intermittent claudication and amputation rate in the RXC group was 20% and 5 %, being lower than the respective rate (56% and 17%) in the control group (P < 0.05); the patency rate was 86% in the RXC group, being higher than that in the control group (65%, P<0.05). Six and 12 months after operation, AAI was 0.73 +/- 0.24 and 0.69 +/- 0.19 respectively in the RXC group, being significantly higher than that in the control group (0.45 +/- 0.17 and 0.41 +/- 0.23, P < 0.05) at the corresponding time points., Conclusion: RXC could obviously increase the patency rate 12 months after FPB, improve the clinical symptoms and alleviate the symptom of limb ischemia in ASO patients.

In our previous study, we found a new mixed formula of Chinese herbs containing Shin-yi-san + Xiao-qing-long-tang + Xiang-sha-liu-jun-zi-tang (9 + 3 + 3 g divided in three doses/day) was beneficial to the patients with perennial allergic rhinitis (AR) via complicated immunomodulatory effects on both mononuclear cells (MNC) and polymorphonuclear neutrophils (PMN). In the present study, we further determined the effects of nasal fluid from AR patients on the functions of human PMN before and after treatment with the mixed formula. We found the nasal discharge, but not serum, from AR group with high serum IgE (H-IgE, serum IgE >200 KIU/l) before treatment exerted many stimulating effects on normal PMN including delayed apoptosis, enhanced production of soluble intercellular adhesion molecule 1 (sICAM-1), interleukin 8 (IL-8) and prostaglandin E2 (PGE2), increased phagocytosis, and augmented cyclooxygenase 2 (COX-2) mRNA expression of PMN. However, these stimulating effects of nasal fluid on PMN were not found in low IgE group (L-IgE, serum IgE <200 KIU/l). These PMN-enhancing effects of H-IgE nasal fluid were abolished after 3-month treatment with the mixed Chinese herb formula. In conclusion, our results suggest that the new mixed herb formula treatment suppressed nasal mucosa inflammation by normalizing stimulatory effects of allergic nasal discharge of patients with H-IgE allergic rhinitis.

A new mixed formula of Chinese herbs containing Shin-yi-san + Xiao-qing-long-tang + Xiang-sha-liu-jun-zi-tang by the weight of 9 + 3 + 3 g/day was prescribed for the treatment of patients with perennial allergic rhinitis for 3 months (the composition of each herb is shown in the tables of the article). We classified the patients into high (H-IgE) and low IgE (L-IgE) groups according to the titer of serum total IgE (> 200 KIU/l in H-IgE vs. < 200 KIU/l in L-IgE) and the presence of house dust mite-specific IgE. The nasal symptomatic score in the high IgE group was significantly improved from 7.19 +/- 0.18 before treatment to 2.67 +/- 0.37 after treatment. In addition, the serum total and house dust mite-specific IgE level were also decreased after treatment. For elucidating the working mechanism of the mixed formula, the Th1 (IFN-gamma) and Th2 (IL-4, IL-5, IL-10 and IL-13) cytokine production by phytohemagglutinin (PHA)-stimulated mononuclear cells (2 x 10(6) cells/ml) and cyclooxygenase type 2 (COX-2) mRNA expression in LPS or IL-13-stimulated PMN were compared before and after 3 months of treatment. We found that the mixed formula treatment significantly enhanced IL-10 but decreased IFN-gamma and IL-5 production by PHA-stimulated mononuclear cells. The IL-5 production was also decreased by PHA-stimulated lymphocyte. In addition, the COX-2 mRNA expression in stimulated PMN was significantly suppressed after treatment. These results suggest that the new mixed formula treatment is beneficial to the patients with perennial allergic rhinitis via modulating the function of lymphocytes and neutrophils.

3,4,5-Trihydroxystibene-3-beta-mono-D-glucoside (PD) from Polygonum cuspidatum 0.05-0.45 mmol/L increased beat rate of myocardial cells from 100% to 112-220%. This action of PD 0.15 mmol/L was agonized by nisoldipine 0.1, propranolol 10 and phentolamine 1 mumol/L. PD (0.05, 0.15 mmol/L) decreased the lactate dehydrogenase (LDH) amount liberated by myocardial cells, from 98 +/- 11 IU/ml of deprivation of oxygen and glucose (DOG) to 55 +/- 16, 68 +/- 14 IU/ml (P less than 0.01) 6 h after injury of DOG; 77 +/- 9, 80 +/- 6 IU/ml (P less than 0.01) 9 h after injury of DOG. PD 0.15 mmol/L decreased the LDH amount liberated by myocardial cells, from 311 +/- 34 to 231 +/- 42 IU/ml (P less than 0.05) 9 h after injury of chlorpromazine (CPZ). It is suggested that PD can protect myocardial cells injured by DOG and CPZ.

The effects of total saponins of Semen Ziziphi spinosae (ZS) were observed on cultured neonatal rat myocardial cells injured by deprivation of oxygen and glucose (DOG) or treatment with chlorpromazine (CPZ) and mitomycin C (MMC). The increases of lactate dehydrogenase release from damaged myocardial cells induced by DOG, CPZ and MMC were attenuated by ZS (33 micrograms/ml) except 11 micrograms/ml which showed no effect on MMC (24 h) and CPZ (9 h)-induced injuries. These data suggest that ZS is an effective protective drug for myocardial cells.