First Report of Leaf Spot Caused by Nigrospora oryzae on Dendrobium candidum in China.

Dendrobium (Dendrobium candidum Wall. ex Lindl.) is a perennial herb in the Orchidaceae family. It has been used as traditional medicinal plant in China, Malaysia, Laos, and Thailand (2). Fungal disease is one of the most important factors affecting the development of Dendrobium production. During summer 2012, chocolate brown spots were observed on leaves of 2-year-old Dendrobium seedlings in a greenhouse in Hangzhou, Zhejiang Province, China, situated at 30.26°N and 120.19°E. Approximately 80% of the plants in each greenhouse were symptomatic. Diseased leaves exhibited irregular, chocolate brown, and necrotic lesions with a chlorotic halo, reaching 0.8 to 3.2 cm in diameter. Affected leaves began to senesce and withered in autumn, and all leaves of diseased plants fell off in the following spring. Symptomatic leaf tissues were cut into small pieces (4 to 5 mm long), surface-sterilized (immersed in 75% ethanol for 30 s, and then 1% sodium hypochlorite for 60 s), rinsed three times in sterilized distilled water, and then cultured on potato dextrose agar (PDA) amended with 30 mg/liter of kanamycin sulfate (dissolved in ddH2O). Petri plates were incubated in darkness at 25 ± 0.5°C, and a grey mycelium with a white border developed after 4 days. Fast-growing white mycelia were isolated from symptomatic leaf samples, and the mycelia became gray-brown with the onset of sporulation after 5 days. Conidia were unicellular, black, elliptical, and 11.4 to 14.3 µm (average 13.1 µm) in diameter. Based on these morphological and pathogenic characteristics, the isolates were tentatively identified as Nigrospora oryzae (1). Genomic DNA was extracted from a representative isolate F12-F, and a ~600-bp fragment was amplified and sequenced using the primers ITS1 and ITS4 (4). BLAST analysis showed that F12-F ITS sequence (Accession No. KF516962) had 99% similarity with the ITS sequence of an N. oryzae isolate (JQ863242.1). Healthy Dendrobium seedlings (4 months old) were used in pathogenicity tests under greenhouse conditions. Leaves were inoculated with mycelial plugs (5 mm in diameter) from a 5-day-old culture of strain F12-F, and sterile PDA plugs served as controls. Seedlings were covered with plastic bags for 5 days and maintained at 25 ± 0.5°C and 80 ± 5% relative humidity. Eight seedlings were used in each experiment, which was repeated three times. After 5 days, typical chocolate brown spots and black lesions were observed on inoculated leaves, whereas no symptoms developed on controls, which fulfilled Koch's postulates. This shows that N. oryzae can cause leaf spot of D. candidum. N. oryzae is a known pathogen for several hosts but has not been previously reported on any species of Dendrobium in China (3). To our knowledge, on the basis of literature, this is the first report of leaf spot of D. candidum caused by N. oryzae in China. References: (1) H. J. Hudson. Trans. Br. Mycol. Soc. 46:355, 1963. (2) Q. Jin et al. PLoS One. 8(4):e62352, 2013. (3) P. Sharma et al. J. Phytopathol. 161:439, 2013. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Sorafenib combined with transarterial chemoembolization for the treatment of advanced hepatocellular carcinoma: a large-scale multicenter study of 222 patients.

BACKGROUND: Data on the efficacy and safety of sorafenib in combination with transarterial chemoembolization (TACE) in patients with advanced hepatocellular carcinoma (HCC) are lacking. PATIENTS AND METHODS: In this multicenter retrospective study, 222 consecutive HCC patients receiving combination therapy were enrolled between June 2008 and July 2011. RESULTS: Chronic hepatitis B was the predominant cause of HCC (86%). Eighty percent patients were at Barcelona Clinic Liver Cancer (BCLC) stage C, and 86% patients were in Child-Pugh (CP) A class. The overall median survival was 12 months (95% CI 10.1-13.9). The overall incidence of adverse events (AEs) was 87%. In 177 BCLC-C patients, performance status, the number of HCC nodules, Child-Pugh score and macrovascular invasion were significantly associated with overall survival (OS) and were included in the final risk scores (R), where R = 5 × (vascular invasion: 0 if no, 1 yes) + 6 × (CP: 0 if A, 1 if B) + 7 × (no. of lesions: 0 if 1-2, 1 ≥3) + 8 × ( Eastern Cooperative Oncology Group, ECOG: 0 if 0, 1 ≥1). CONCLUSIONS: Sorafenib in combination with TACE should be considered a safe and effective therapy for advanced HCC. Further validation of the new subgroup of BCLC-C stage is warranted in an independent patient cohort.

Kinsenoside prevents ovariectomy-induced bone loss and suppresses osteoclastogenesis by regulating classical NF-κB pathways.

UNLABELLED: Kinsenoside is able to improve bone turnover rate in ovariectomized (OVX) mice. In vitro analysis shows that kinsenoside antagonizes osteoclast development and bone resorption. INTRODUCTION: Kinsenoside, the main active compound of the traditional Taiwanese herb Anoectochilus formosanus, has an antiinflammatory effect. This study investigates whether kinsenoside inhibits osteoporosis and osteoclastogenesis. METHODS: OVX mice were used to examine the antiosteoporotic activity of kinsenoside. The trabecular bone microarchitecture was assessed by microcomputed tomography. In vitro experiments were performed to determine the mechanisms of the antiosteoporotic effects of kinsenoside. RESULTS: Microcomputed tomography scanning showed that kinsenoside suppresses bone loss in OVX mice. Kinsenoside decreases plasma CTx concentration. Reverse transcription polymerase chain reaction (RT-PCR) analysis also showed that kinsenoside reduces the femoral mRNA expression of tartrate-resistant acid phosphatase (TRAP) and matrix metalloproteinase-9 (MMP-9). Kinsenoside inhibits osteoclast formation in bone marrow cells (BMs) and RAW 264.7 cells. Western blot was used to analyze osteoclast-associated signaling pathways in RAW 264.7 cells. Results show that kinsenoside does not inhibit IKK phosphorylation but suppresses the phosphorylation of IκBα and p65. Kinsenoside significantly inhibits the RANKL induction of IKK activity. Kinsenoside inhibits the RANKL-triggered nuclear translocations of NF-κB and nuclear factor of activated T cells c1 (NFATc1). RT-PCR was used to analyze osteoclast precursor fusion and resorption-associated gene expression in BMs. Kinsenoside inhibits the expression of cathepsin K (CAK), dendritic cell-specific transmembrane protein, MMP-9, and TRAP. CONCLUSIONS: Kinsenoside inhibits osteoclastogenesis from macrophages by attenuating RANKL-induced NF-κB and NFATc1 activities, which in turn, prevents bone loss from OVX mice.

Potent antiplatelet, anti-inflammatory and antiallergic isoflavanquinones from the roots of Abrus precatorius.

Five isoflavanquinones have been isolated from the roots of Abrus precatorius L. (Leguminosae). Three of them are new and designated as abruquinones D, E, and F. The pharmacological activities of the isoflavanquinones have been evaluated. The results indicated that abruquinones A, B, and D exhibited remarkable inhibitory effects on the platelet aggregation. The IC50 of abruquinones A and B for the inhibition of the platelet aggregation induced by arachidonic acid (AA) and collagen were less than 5 micrograms/ml, and of abruquinone D, was less than 10 micrograms/ml for that induced by AA. On the other hand, abruquinones A, B, D, and F showed strong anti-inflammatory and antiallergic effects. The IC50 of abruquinones A, B, D, and F for the inhibition of superoxide formation were less than 0.3 micrograms/ml, for the inhibition of the release of both beta-glucuronidase and lysozyme from rat neutrophils and the release of both beta-glucuronidase and histamine from mast cells were less than 1 microgram/ml.

Platelet activating factor (PAF) antagonists contained in medicinal plants: lignans and sesquiterpenes.

Hot aqueous extracts of medicinal plants were tested for their inhibitory effect on the binding of platelet activating factor (PAF) to rabbit platelets. The extracts of Forsythia suspensa VAHL. (Oleaceae), Arctium lappa L. (Compositae) and Centipeda minima (L.) A. BRAUN et ASCHERS (Compositae) showed significant activities. Since the main constituents of F. suspensa and A. lappa are lignans, 30 lignans were tested for their inhibitory effects on PAF binding to platelets and 9 lignans were found active. Four sesquiterpenes were isolated as active compounds from C. minima. In particular 6-O-angeloylplenolin and 6-O-senecioyplenolin are the most potent and specific PAF antagonists found in this study.

Biologically active constituents of Centipeda minima: sesquiterpenes of potential anti-allergy activity.

Ether, methanol and aqueous extracts of Centipeda minima (Compositae) herbs were found to have significant anti-allergy activities in passive cutaneous anaphylaxis (PCA) test. Three flavonoids, two sesquiterpene lactones and an amide were isolated from this plant material as inhibitors to induced histamine release from mast cells. The sesquiterpenes were identified as isobutyroylplenolin and senecioylplenolin by spectral investigations. The flavonoids and sesquiterpenes exhibited significant anti-allergy activity in PCA test with p.o. administration.

6-Pentadecylsalicylic acid: an antithrombin component isolated from the stem of Rhus semialata var. roxburghii.

Bioassay-directed fractionation of the n-hexane extract of the stem of Rhus semialata Murr. var. roxburghii DC (Anacardiaceae) has led to the isolation of 6-pentadecylsalicylic acid. It showed antithrombin activity at 50 micrograms/ml in the amidolytic method. It also prolonged the clotting time in a dose-dependent manner in the clotting assay of thrombin-fibrinogen interaction.