Comparative study of Scleromitrion diffusum and Oldenlandia corymbosa: Microscopy, TLC, HPLC, and antioxidant activity.

Quality control of herbal medicines is crucial, especially the role of herbal drug identification. This is essential for preventing the misuse of herbs, which can affect efficacy or cause toxicity. Scleromitrion diffusum is a common herb, yet it is often mistaken for Oldenlandia corymbosa. This study analyzed the morphology, microscopy, thin-layer chromatography (TLC), and high-pressure liquid chromatography (HPLC) using two markers, asperuloside and scandoside methyl ester, to distinguish between S. diffusum and O. corymbosa with the analysis included 10 samples of S. diffusum and 10 samples of O. corymbosa collected from the Taiwan market. By quantifying the total polyphenols and flavonoids, we investigated the antioxidant activity, including the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect, 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) scavenging effect, and reducing power to further elucidate the biological effects of the two herbs. The results of this study revealed notable differences in microscopy and suggested a TLC method for distinguishing between the two herbs in the market. In HPLC, the ratios of asperuloside and scandoside methyl ester differed between the two herbs. S. diffusum contained a higher asperuloside content. In contrast, O. corymbosa contained higher concentrations of scandoside methyl esters. With more total polyphenols and flavonoids in S. diffusum than those in O. corymbosa, the antioxidant activity of S. diffusum was superior to that of O. corymbosa. This study provides a comprehensive understanding for the identification and quality evaluation of S. diffusum in the market. RESEARCH HIGHLIGHTS: The study consolidates and clarifies the morphological and microscopic differences between Scleromitrion diffusum and Oldenlandia corymbosa - a common adulterant species of S. diffusum on the Taiwan markets. Using Asperuloside and Scandoside methyl ester as two chemical markers, the study proposes a TLC method for rapidly testing S. diffusum and O. corymbosa on the market. Through HPLC analysis, our results showed that S. diffusum and O. corymbosa had a clear difference in the ratio of two markers, Asperuloside and Scandoside methyl ester: Asperuloside/Scandoside methyl ester in S. diffusum is higher than that in O. corymbosa. Through phytochemicals contents, including total phenols content, flavonoids content, and antioxidant activity, including DPPH, ABTS•+ scavenging activity, and reducing power, S. diffusum showed slightly higher levels of phenols and flavonoids as well as a better antioxidant activity than O. corymbosa.

Qualitative and quantitative analysis methods for quality control of rhubarb in Taiwan's markets.

Introduction: Rhubarb is a traditional Chinese medicine (TCM) used to release heat and has cathartic effects. Official rhubarb in Taiwan Herbal Pharmacopeias 4th edition (THP 4th) and China Pharmacopeia 2020 (CP 2020) are the roots and rhizomes of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf., and Rheum officinale Baill. However, the Rheum genus is a large genus with many different species, and owing to the similarity in appearance and taste with official rhubarb, there needs to be more clarity in the distinction between the species of rhubarb and their applications. Given the time-consuming and complicated extraction and chromatography methods outlined in pharmacopeias, we improved the qualitative analysis and quantitative analysis methods for rhubarb in the market. Hence, we applied our method to identify the species and quality of official and unofficial rhubarb. Method: We analyzed 21 rhubarb samples from the Taiwanese market using a proposed HPLC-based extraction and qualitative analysis employing eight markers: aloe-emodin, rhein, emodin, chrysophanol, physcion, rhapontigenin, rhaponticin, and resveratrol. Additionally, we developed a TLC method for the analysis of rhubarb. KEGG pathway analysis was used to clarify the phytochemical and pharmacological knowledge of official and unofficial rhubarb. Results: Rhein and rhapontigenin emerged as key markers to differentiate official and unofficial rhubarb. Rhapontigenin is abundant in unofficial rhubarb; however, rhein content was low. In contrast, their contents in official rhubarb were opposite to their contents in unofficial rhubarb. The TLC analysis used rhein and rhapontigenin to identify rhubarb in Taiwan's markets, whereas the KEGG pathway analysis revealed that anthraquinones and stilbenes affected different pathways. Discussion: Eight reference standards were used in this study to propose a quality control method for rhubarb in Taiwanese markets. We propose a rapid extraction method and quantitative analysis of rhubarb to differentiate between official and unofficial rhubarb.

High-dose Tiger-Gian formula protects the knee joint from surgically induced osteoarthritis in rats.

AIM: Although the Tiger-Gian formula (TGF) has proven clinically effective at improving the symptoms of knee osteoarthritis (KOA), the pharmacological effects and underlying mechanisms of TGF have not been examined in any animal model. This study assessed the effects of TGF in male Sprague-Dawley rats with anterior cruciate ligament transection (ACLT) -induced KOA. METHODS: Thirty rats underwent ACLT surgery and were assigned to either the control group, ACLT alone, ACLT + low-dose TGF (1000 mg/kg), ACLT + high-dose TGF (3000 mg/kg), or ACLT + celecoxib (30 mg/kg). All rats were subjected to micro-computed tomography (micro-CT), weight-bearing behavioral testing, and histological inspections of the knee joint for evidence of structural changes in articular bone, cartilage and synovium. RESULTS: After 6 weeks, force discrepancies in weight-bearing distribution between the normal hind and postoperative limbs revealed superiority with high-dose TGF (18.00 ± 5.93 g) and celecoxib (18.68 ± 5.29 g) versus both ACLT alone (41.29 ± 7.06 g) and low-dose TGF (37.00 ± 7.40 g). Micro-CT images revealed that high-dose TGF and celecoxib similarly improved subchondral bone architecture, protected articular cartilage after ACLT, and downregulated proinflammatory cytokines interleukin-1ß and tumor necrosis factor-α in the cartilage and synovial sections. CONCLUSION: High-dose TGF induced the smallest amount of KOA-associated bone loss. Anti-inflammatory, anti-oxidative, and immunomodulatory effects of TGF were accompanied by reductions in proinflammatory cytokines and improvements in pain and function. TGF-induced anti-osteoporotic activity and inhibition of cartilage degradation were reflected by micro-CT and histological analysis. The findings help to explain how TGF alleviates symptoms of KOA.

Characterization of chemical constituents with their antioxidant and anti-melanogenesis activities from the roots of Ampelopsis japonica.

Three glycosylated stilbenes (1-3), two anthraquinones (4, 5), one lignan (6), five tannins (7-11), two amino acids (12, 13), and one auronol (14) were isolated from the root of Ampelopsis japonica. All compounds, except for 4, 6, and 11 were obtained from this species for the first time. Compounds 6-9 could notably inhibit ROS generations in HaCaT keratinocyte cells with IC50 values of 5.28, 4.83, 0.87, and 1.66 µM, respectively. Compounds 8-10 showed potent DPPH free radical scavenging effects with IC50 values of 14.37, 16.08, and 12.11 µM, individually. In anti-melanogenesis assay, only 8 and 9 could decrease 7.93% and 11.66% melanin contents induced by α-MSH in B16F10 melanoma cells at 40 µM and moderately inhibit tyrosinase activities. By far, galloylhamameloses 8 and 9 were found to exhibit both antioxidant and anti-melanogenesis properties that could be further developed as cosmeceutical agents for skin disorders.

Tuliposides H-J and Bioactive Components from the Bulb of Amana edulis.

Three new tuliposides H-J (1-3) and 11 known compounds were obtained from the methanolic extracts of the bulbs of Amana edulis for the first time. Their structures were elucidated by NMR, MS, and IR spectroscopic data, optical rotation, and Mosher's method. The melanogenesis properties of all the isolates were evaluated in B16 melanoma cells. Consequently, tributyl citrate (9) had anti-melanogenesis activity but was cytotoxic toward B16. (+)-Pyroglutamic acid (4), (+)-butyl 5-oxopyrrolidine-2-carboxylate (6), (-)-3-hydroxy-2-methylbutyrolactone (10), and 5-(hydroxymethyl)furfural (12) had increased melanin productions and tyrosinase activities. Those active components could be further studied as the candidates against melanoma and vitiligo for skin diseases or whitening/hypopigmentation for hair.

Interactive Association Between Intronic Polymorphism (rs10506151) of the LRRK2 Gene and Type 2 Diabetes on Neurodegenerative Diseases.

PURPOSE: We investigated the interactive effect of rs10506151 polymorphism of the Leucine-rich repeat kinase 2 (LRRK2) gene and type 2 diabetes (T2D) on neurodegenerative disease (ND) risk. MATERIALS AND METHODS: Data of 17, 927 participants in the Taiwan Biobank (TWB) assessed between 2008 and 2015 were linked to healthcare records in the National Health Insurance Research Database (NHIRD). The odd ratios (ORs) and 95% confidence intervals (CIs) for NDs were determined using logistic regression analysis. RESULTS: There were 145 cases with NDs, and 28.28% (n = 41) of these individuals had T2D. Associations of neurodegenerative disorders with LRRK2 rs10506151 variant and T2D were not significant. The corresponding ORs (95% CI) for NDs were 1.06 (0.75-1.49) in CA/AA compared to CC individuals and 0.93 (0.63-1.39) in those with T2D compared to non-diabetic participants. However, we found evidence of a significant interaction between rs10506151 and T2D (p = 0.0073). After stratification by genotypes of rs10506151, the OR for NDs was 0.37 (CI, 0.17-0.82) in CA/AA individuals with T2D and 1.41 (0.88-2.27) in their CC counterparts. When CA/AA individuals with T2D represented the reference group, the OR (95% CI) was 1.74 (0.81-3.73) in CC individuals with no T2D, 2.47 (CI, 1.14-5.38) in CA/AA individuals with no T2D, and 2.34 (CI, 1.07-5.11) in CC individuals with T2D. CONCLUSION: Our data indicated that the risk of NDs was significantly lower among diabetic individuals with combined CA/AA of the LRRK2 rs10506151 variant in Taiwan.

Imperatorin Interferes with LPS Binding to the TLR4 Co-Receptor and Activates the Nrf2 Antioxidative Pathway in RAW264.7 Murine Macrophage Cells.

Imperatorin (IMP) could downregulate several inflammatory transcription factor signaling pathways. Some studies have pointed out that IMP could interfere with toll-like receptor 4 (TLR4) signaling. This study evaluates how IMP interferes with the TLR4 co-receptors signaling through the protein-ligand docking model, Western blotting, immunofluorescence (IF), and atomic force microscopy (AFM) assays in lipopolysaccharide (LPS) stimulated macrophage-like RAW264.7 cells in vitro. The results of the protein-ligand docking demonstrate that IMP interferes with LPS binding to the LPS-binding protein (LBP), the cluster of differentiation 14 (CD14), and the toll-like receptor 4/myeloid differentiation factor 2 (TLR4/MD-2) co-receptors in LPS-stimulated RAW264.7 cells. Compared with TLR4 antagonist CLI-095 or dexamethasone, IMP could suppress the protein expressions of LBP, CD14, and TLR4/MD-2 in LPS-stimulated cells. Furthermore, the three-dimensional (3D) image assay of the AFM showed IMP could prevent the LPS-induced morphological change in RAW264.7 cells. Additionally, IMP could activate the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, and it increased the antioxidative protein expression of heme oxygenase-1 (HO-1), superoxidase dismutase (SOD), and catalase (CAT). Our results are the first to reveal that the anti-inflammatory effect of IMP interferes with LPS binding to TLR4 co-receptor signaling and activates the antioxidative Nrf2 signaling pathway.

New ent-kauran diterpene and antioxidant components from the seed of Ipomoea nil.

One new ent-kauran diterpene, 7ß,16ß,17-trihydroxy ent-kauran 19-(6ß)-olide (1), along with eight known compounds were isolated from the seed of Ipomoea nil. Isolates caffeoylquinic acid derivatives 5-9 were found for the first time in this species. All structures were identified from various spectroscopic data. trans-Caffeic acid 3, phenylpropanoid 4, and caffeoylquinic acid derivatives 5-9 could inhibit ROS generations induced in human keratinocyte HaCaT cells with IC50 values of 0.94-28.40 µM. Compounds 3 and 5-9 also had DPPH free radical scavenging properties (IC50 values, 14.86-68.27 µM), however, isolate 4 did not show inhibition effect. Generally, I. nil and its secondary metabolites 3-9 could be further applied for oxidative stress damage resulted in skin disorders.

Novel Caryophyllane-Related Sesquiterpenoids with Anti-Inflammatory Activity from Rumphella antipathes (Linnaeus, 1758).

Two previously undescribed caryophyllane-related sesquiterpenoids, antipacids A (1) and B (2), with a novel bicyclo[5.2.0] core skeleton, and known compound clovane-2ß,9α-diol (3), along with rumphellolide L (4), an esterified product of 1 and 3, were isolated from the organic extract of octocoral Rumphella antipathes. Their structures, including the absolute configurations were elucidated by spectroscopic and chemical experiments. In vivo anti-inflammatory activity analysis indicated that antipacid B (2) inhibited the generation of superoxide anions and the release of elastase by human neutrophils, with IC50 values of 11.22 and 23.53 µM, respectively, while rumphellolide L (4) suppressed the release of elastase with an IC50 value of 7.63 µM.

Antiviral activity of Portulaca oleracea L. against influenza A viruses.

ETHNOPHARMACOLOGICAL RELEVANCE: Portulaca oleracea L. is used not only as an edible potherb but also as a traditional remedy to assuage the symptoms of various diseases. The water extract of P. oleracea (WEPO) has been found to effectively alleviate the signs and symptoms of pandemic influenza A virus (IAV) infection. However, the anti-IAV activity of WEPO is still unclear. AIM OF STUDY: In this study, we aimed to elucidate the anti-IAV activity of WEPO and investigate the potential mechanisms underlying the anti-H1N1 activity. MATERIALS AND METHODS: The cytotoxicity of WEPO and other Chinese herbs was measured using the cell viability test. The anti-IAV activity of WEPO was determined using the plaque reduction assay, real-time reverse transcription-polymerase chain reaction, and immunofluorescence assay. The virucidal activity of WEPO was determined by labeling the virus and using the time-dependent virucidal activity assay. RESULTS: The half-maximal effective concentration of WEPO for A/WSN/1933 (H1N1) was very low, with a high selectivity index. The production of circulating H1N1 and H3N2 was suppressed by WEPO. Additionally, the antiviral activity of WEPO was observed in the early stage of IAV infection. Furthermore, WEPO inhibited the binding of virus to cells and exhibited good virucidal activity, significantly decreasing the viral load within 10 min to prevent viral infection. CONCLUSIONS: We demonstrate the anti-IAV activity of WEPO and strongly recommend the use of WEPO, as an herbal regimen, to prevent and treat H1N1 infection at an early stage.

Cis-3-O-p-hydroxycinnamoyl Ursolic Acid Induced ROS-Dependent p53-Mediated Mitochondrial Apoptosis in Oral Cancer Cells.

Cis-3-O-p-hydroxycinnamoyl ursolic acid (HCUA), a triterpenoid compound, was purified from Elaeagnus oldhamii Maxim. This traditional medicinal plant has been used for treating rheumatoid arthritis and lung disorders as well as for its anti-inflammation and anticancer activities. This study aimed to investigate the anti-proliferative and apoptotic-inducing activities of HCUA in oral cancer cells. HCUA exhibited anti-proliferative activity in oral cancer cell lines (Ca9-22 and SAS cells), but not in normal oral fibroblasts. The inhibitory concentration of HCUA that resulted in 50% viability was 24.0 µM and 17.8 µM for Ca9-22 and SAS cells, respectively. Moreover, HCUA increased the number of cells in the sub-G1 arrest phase and apoptosis in a concentration-dependent manner in both oral cancer cell lines, but not in normal oral fibroblasts. Importantly, HCUA induced p53-mediated transcriptional regulation of pro-apoptotic proteins (Bax, Bak, Bim, Noxa, and PUMA), which are associated with mitochondrial apoptosis in oral cancer cells via the loss of mitochondrial membrane potential. HCUA triggered the production of intracellular reactive oxygen species (ROS) that was ascertained to be involved in HCUA-induced apoptosis by the ROS inhibitors YCG063 and N-acetyl-L-cysteine. As a result, HCUA had potential antitumor activity to oral cancer cells through eliciting ROS-dependent and p53-mediated mitochondrial apoptosis. Overall, HCUA could be applicable for the development of anticancer agents against human oral cancer.

Chrysin inhibit human melanoma A375.S2 cell migration and invasion via affecting MAPK signaling and NF-κB signaling pathway in vitro.

Numerous evidences have shown that chrysin induced cytotoxic effects via induced cell cycle arrest and induction of cell apoptosis in human cancer cell lines, however, no information showed that chrysin inhibited skin cancer cell migration and invasion. In this study, we investigated anti-metastasis mechanisms of chrysin in human melanoma cancer A375.S2 cells in vitro. Under sub-lethal concentrations of chrysin (0, 5, 10, and 15 µM) which inhibits cell mobility, migration and invasion of A375.S2 cells that were assayed by wound healing and Transwell filter. That chrysin inhibited MMP-2 activity in A375.S2 cells was investigated by gelatin zymography assay. Western blotting was used to examine protein expression and results indicated that chrysin inhibited the expression of GRB2, SOS-1, PKC, p-AKT (Thr308), NF-κBp65, and NF-κBp50 at 24 and 48 hours treatment, but only at 10-15 µM of chrysin decreased Ras, PI3K, p-c-Jun, and Snail only at 48 hours treatment and only decrease p-AKT(Ser473) at 24 hours treatment. Furthermore, chrysin (5-15 µM) decreased the expression of uPA, N-cadherin and MMP-1 at 24 and 48 hours treatment but only decreased MMP-2 and VEGF at 48 hours treatment at 10-15 µM and 5-15 µM of chrysin, respectively, however, increased E-cadherin at 5-15 µM treatment. Results of confocal laser microscopy systems indicated that chrysin inhibited expression of NF-κBp65 in A375.S2 cells. Based on these observations, we suggest that chrysin can be used in anti-metastasis of human melanoma cells in the future.

Hepatoprotective Effect of Ugonin M, A Helminthostachyszeylanica Constituent, on Acetaminophen-Induced Acute Liver Injury in Mice.

The present study aimed to discover the possible effectiveness of Ugonin M, a unique flavonoid isolated from Helminthostachys zeylanica-a traditional Chinese medicine used as anti-inflammatory medicine-and to elucidate the potential mechanisms of Ugonin M in the acute liver injury induced by acetaminophen (APAP). In this study, Ugonin M significantly ameliorated APAP-induced histopathological changes and the typical liver function biomarkers (i.e., alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (T-Bil)). It also affected APAP-induced abnormal lipid metabolism including total cholesterol (TC) and triglyceride (TG) in the serum. In inflammatory pharmacological action, Ugonin M suppressed the pro-inflammatory mediators such as nitric oxide (NO) and the lipid peroxidation indicator malondialdehyde (MDA). In addition, Ugonin M reinforced hemeoxygenase-1 (HO-1) protein expression and the production of antioxidant enzymes viz superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). Furthermore, inflammation-associated cytokines including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1ß as well as proteins such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were decreased by the pretreatment of Ugonin M. Moreover, this study found that pretreatment of Ugonin M apparently decreased nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) activation via inhibition of the degradation of NF-κB, inhibitory κB-α (IκB-α), extracellular regulated kinase (ERK), c-Jun-N-terminal (JNK), and p38 active phosphorylation. In conclusion, Ugonin M significantly showed a protective effect against APAP-induced liver injury by reducing oxidative stress and inflammation. Thus, Ugonin M could be one of the effective components of H. zeylanica that plays a major role in the treatment of inflammatory disorders.

Ginsenoside Rh2 Ameliorates Lipopolysaccharide-Induced Acute Lung Injury by Regulating the TLR4/PI3K/Akt/mTOR, Raf-1/MEK/ERK, and Keap1/Nrf2/HO-1 Signaling Pathways in Mice.

The anti-inflammatory effect of ginsenoside Rh2 (GRh2) has labeled it as one of the most important ginsenosides. The purpose of this study was to identify the anti-inflammatory and antioxidant effects of GRh2 using a lipopolysaccharide (LPS) challenge lung-injury animal model. GRh2 reduced LPS-induced proinflammatory mediator nitric oxide (NO), tumor necrosis factor-alpha, interleukin (IL)-1ß, and anti-inflammatory cytokines (IL-4, IL-6, and IL-10) production in lung tissues. GRh2 treatment decreased the histological alterations in the lung tissues and bronchoalveolar lavage fluid (BALF) protein content; total cell number also reduced in LPS-induced lung injury in mice. Moreover, GRh2 blocked iNOS, COX-2, the phosphorylation of IκB-α, ERK, JNK, p38, Raf-1, and MEK protein expression, which corresponds with the growth of HO-1, Nrf-2, catalase, SOD, and GPx expression in LPS-induced lung injury. An in vivo experimental study suggested that GRh2 has anti-inflammatory effects, and has potential therapeutic efficacy in major anterior segment lung diseases.

New diterpenes leojaponins G-L from Leonurus japonicus.

Six new diterpenes, leojaponins G-L (1-6) along with 19 known compounds (7-25) were isolated from Leonurus japonicus. Their structures were elucidated by NMR, MS, IR, UV, and ECD spectroscopic data. Anti-melanogenesis assay indicated that 7 could safely and dose-dependently decrease melanin production in B16F10 melanoma cell with an IC50 value of 59.1 µM, but moderately inhibit tyrosinase activity. Without cytotoxicities at 20 µM, compounds 11, 14, 15, and 17-21 showed significant melanogenesis stimulation activities at the percentages of 7.7-48.2. Antioxidants 19 and 24 could notably inhibit ROS production in a dose-dependent manner with percentages of 24.7-42.2 and 27.9-40.2, respectively among the concentrations of 0.16 to 100 µM. Our results demonstrated L. japonicus and its constituents could be potential botanical resources of cosmeceutical development for treatment and prevention of skin disorders.

Quality Control of the Root and Rhizome of Helminthostachys zeylanica (Daodi-Ugon) by HPLC Using Quercetin and Ugonins as Markers.

Daodi-Ugon is the dried root and rhizome of Helminthostachys zeylanica (L.) Hook. and has been used for centuries in the treatment of inflammation, fever, pneumonia, burns, and various disorders. However, the chromatographic methods to determine the phytochemical composition of H. zeylanica have never been reported. This study not only aims to develop a valid high-performance liquid chromatography (HPLC) method and to establish a chromatographic fingerprint for the quality control of H. zeylanica, it also establish the proposed content limits of Quercetin, Ugonin J, and Ugonin M. An HPLC method with a RP18 column (250 × 4.6 mm, 5 µm) was developed for the quantitative analysis of Quercetin, Ugonin J, and Ugonin M in H. zeylanica. A simple gradient of (A) methanol/(B) phosphoric acid in water (5-45 min, 70-80% A; 50-55 min, 80-70% A) was used and 360 nm was selected as the detection wavelength. The average contents and proposed content limits for H. zeylanica were calculated with a t-test and a measurement uncertainty test based on 20 batches of authentic H. zeylanica samples. Limits of detection (LOD), quantification (LOQ), linearity, precision, repeatability, stability, and recovery of the developed method were validated. All of the validation results of quantitative determination and fingerprinting methods were satisfactory. The developed method was then applied to assay the contents of Quercetin, Ugonin J, and Ugonin M and to acquire the fingerprints of all of the collected H. zeylanica samples. At the 99% confidence level, the calculated content limits were 56.45, 112.15, and 277.98 mg/kg for Quercetin, Ugonin J, and Ugonin M, respectively. Those validated HPLC quantitative method, fingerprinting profile, and the proposed content limits of three chemical markers that could be used in the quality control of H. zeylanica in the market.

Ugonin M, a Helminthostachys zeylanica Constituent, Prevents LPS-Induced Acute Lung Injury through TLR4-Mediated MAPK and NF-κB Signaling Pathways.

Helminthostachys zeylanica (L.) Hook. is plant that has been used in traditional Chinese medicine for centuries for the treatment of inflammation, fever, pneumonia, and various disorders. The aims of the present study are to figure out the possible effectiveness of the component Ugonin M, a unique flavonoid isolated from H. zeylanica, and to elucidate the mechanism(s) by which it works in the LPS-induced ALI model. In this study, Ugonin M not only inhibited the production of pro-inflammatory mediators such as NO, TNF-α, IL-1ß, and IL-6, as well as infiltrated cellular counts and protein content in the bronchoalveolar lavage fluid (BALF) of lipopolysaccharides (LPS)-induced acute lung injury (ALI) mice, but also ameliorated the severity of pulmonary edemas through the score of a histological examination and the ratio of wet to dry weight of lung. Moreover, Ugonin M was observed to significantly suppress LPS-stimulated protein levels of iNOS and COX-2. In addition, we found that Ugonin M not only obviously suppressed NF-κB and MAPK activation via the degradation of NF-κB and IκB-α as well as ERK and p38MAPK active phosphorylation but also inhibited the protein expression level of TLR4. Further, Ugonin M treatment also suppressed the protein levels of MPO and enhanced the protein expressions of HO-1 and antioxidant enzymes (SOD, GPx, and CAT) in lung tissue of LPS-induced ALI mice. It is anticipated that through our findings, there is strong evidence that Ugonin M may exert a potential effect against LPS-induced ALI mice. Hence, Ugonin M could be one of the major effective components of H. zeylanica in the treatment of inflammatory disorders.

Chemical compositions, chromatographic fingerprints and antioxidant activities of Citri Exocarpium Rubrum (Juhong).

BACKGROUND: Citri Exocarpium Rubrum (CER), which is known as Juhong in Chinese, is the dried exocarp of Citrus reticulata Blanco and its cultivars (Fam. Rutaceae) and is currently used in Chinese medicine to protect the stomach and eliminate dampness and phlegm. The main aim of this study was to develop a high-performance liquid chromatography ultraviolet mass spectrometry (HPLC-UV-MS) method for determining the chemical compositions and fingerprint of CER. We also evaluated the antioxidant properties of CER based on its 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, ferric ion reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays. METHODS: Ten CER samples were collected from Hong Kong and mainland China. Each CER sample was extracted using an ultrasonic extraction method. Chromatographic separation was achieved using a conventional Dikma Inspire C18 column with photo diode array detection (190-400 nm). Hesperidin, nobiletin and tangeretin were quantified based on the UV signal observed at 330 nm. The column was eluted with a mobile phase consisting of water and acetonitrile (15-55%) over 55 min. Fingerprints combined with similarity and principal component analyses were used to classify the herbs. The DPPH free radical scavenging activity, FRAP and ABTS properties of the different CER samples were assayed. Bivariate correlation analysis was performed to investigate the correlation between the characteristic peaks and their antioxidant capacities. RESULTS: Limit of detection (LOD), limit of quantification (LOQ), linearity, inter-day precision, intra-day precision, repeatability, stability and recovery of the developed method were validated, and the method was subsequently used to determine the contents of hesperidin, nobiletin and tangeretin, and to acquire the fingerprints of the CER samples. Seventeen characteristic peaks were found in the fingerprints, and eleven of them were identified. Bivariate correlation analysis revealed correlations between the characteristic peaks and the antioxidant activities of the samples. CONCLUSION: An HPLC-UV-MS method was developed and validated after a detailed investigation on extraction of chemical compounds from CER using different solvents and extraction times. None of the peaks was correlated with the DPPH free radical scavenging activity or ferric reducing capacity. Most of the peaks were correlated well with the ABTS radical scavenging capacity.

Utilization Pattern and Drug Use of Traditional Chinese Medicine, Western Medicine, and Integrated Chinese-Western Medicine Treatments for Allergic Rhinitis Under the National Health Insurance Program in Taiwan.

OBJECTIVES: Patients in Taiwan with allergic rhinitis seek not only Western medicine treatment but also Traditional Chinese Medicine treatment or integrated Chinese-Western medicine treatment. Various studies have conducted pairwise comparison on Traditional Chinese Medicine, Western medicine, and integrated Chinese-Western medicine treatments. However, none conducted simultaneous analysis of the three treatments. This study analyzed patients with allergic rhinitis receiving the three treatments to identify differences in demographic characteristic and medical use and thereby to determine drug use patterns of different treatments. MATERIALS AND METHODS: The National Health Insurance Research Database was the data source, and included patients were those diagnosed with allergic rhinitis (International Classification of Diseases, Ninth Revision, Clinical Modification codes 470-478). Chi-square test and Tukey studentized range (honest significant difference) test were conducted to investigate the differences among the three treatments. RESULTS: Visit frequency for allergic rhinitis treatment was higher in female than male patients, regardless of treatment with Traditional Chinese Medicine, Western medicine, or integrated Chinese-Western medicine. Persons aged 0-19 years ranked the highest in proportion of visits for allergic rhinitis. Traditional Chinese Medicine treatment had more medical items per person-time and daily drug cost per person-time and had the lowest total expenditure per person-time. In contrast, Western medicine had the lowest daily drug cost per person-time and the highest total expenditure per person-time. The total expenditure per person-time, daily drug cost per person-time, and medical items per person-time of integrated Chinese-Western medicine treatment lay between those seen with Traditional Chinese Medicine and Western medicine treatments. CONCLUSIONS: Although only 6.82 % of patients with allergic rhinitis chose integrated Chinese-Western medicine treatment, the visit frequency per person-year of integrated Chinese-Western medicine ranked highest. In addition, multiple-composition medicines were used more frequently than single-composition medicines, and mar huang (Ephedra sinica Stapf) was seldom used to decrease the risk of combining medications.

Lobeline improves acute lung injury via nuclear factor-κB-signaling pathway and oxidative stress.

Acute lung injury (ALI) is a severe, life-threatening medical condition whose pathogenesis is linked to neutrophil infiltration of the lung. Activation and recruitment of neutrophils to the lung is mostly attributed to the production of chemokines NO, IL-6, for instance. This study aims to investigate lobeline ability in reducing NO production, and nitric oxide synthase (iNOs) expression. Lobeline was tested by inhibiting phosphorylation of mitogen-activated protein kinases (MAPKs), NF-κB and IκBα in LPS-stimulated RAW 264.7 cells. When RAW 264.7 macrophages were given lobeline with LPS, a significant concentration-dependent inhibition of NO production was detected. In vivo tests, mice were either treated with normal saline, 10mg/kg dexmethasone or 5, 10, 20mg/kg lobeline intraperitoneally, and after an hour, the administration of 5mg/kg of LPS was given intratracheally. External performance, cytokines, MAPK pathways and antioxidative enzymes (AOEs) were also carried out to evaluate the effects of these drugs. This is the first investigation in which lobeline was found to effectively inhibit acute lung edema, which may provide a potential target for treating ALI. Lobeline may utilize MAPKs pathways as well as AOEs activity to attenuate LPS-induced nonspecific pulmonary inflammation.