Effects of Flower Buds Extract of Tussilago farfara on Focal Cerebral Ischemia in Rats and Inflammatory Response in BV2 Microglia / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the flower buds extract of Tussilago farfara Linné (Farfarae Flos; FF) on focal cerebral ischemia through regulation of inflammatory responses in activated microglia.</p><p><b>METHODS</b>Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion (tMCAO) for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups (n=5 per group): normal, tMCAO-induced ischemic control, tMCAO plus FF extract 300 mg/kg-treated, and tMCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract (300 mg/kg, p.o.) or MK-801 (1 mg/kg, i.p.) was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei (NeuN), anti-glial fibrillary acidic protein (GFAP), and anti-CD11b/c (OX42) antibodies in ischemic brain. The expressions of inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF-α), and hypoxia-inducible factor-1a (HIF-1α) were determined by Western blot. BV2 microglial cells were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide (LPS). Nitric oxide (NO) production was measured in culture medium by Griess assay. The expressions of iNOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of iNOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot.</p><p><b>RESULTS</b>FF extract significantly decreased brain infarctions in ischemic rats (P<0.01). The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed iNOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract (0.2 and 0.5 mg/mL, P<0.01) and tussilagone 20 and 50 μmol/L, P<0.01) significantly decreased LPS-induced NO production in BV2 microglia through downregulation of iNOS mRNA and protein expression. FF extract and tussilagone significantly inhibited LPS-induced expression of TNF-α, IL-1β, and IL-6 mRNA, and also suppressed the phosphorylation of ERK1/2, JNK and p38 MAPK and the nuclear expression of NF-κB in a dose-dependent manner.</p><p><b>CONCLUSIONS</b>FF extract has a neuroprotective effect in ischemic stroke by the decrease of brain infarction, and the inhibition of neuronal death and microglial activation-mediated inflammatory responses.</p>

Character Association Studies in M2 Generation of Fieldpea (Pisum sativum var. arvense L.).

An experiment was carried out with an objective to study the association among characters of fieldpea in M2 generation. The parent material, seeds of PUSA212 variety were irradiated with 10kR, 15kR, 20kR, 25kR and 30kR doses of gamma rays at NBRI, Lucknow. Next day after treatment, the seeds along with control were space planted for raising M1 generation. Each M1 plant was harvested separately. Desirable ten M1 individual plant progenies from each treatment were bulked and laid in RBD for rising M2 generation. Induced mutations delivered fairly good amount of genotypic correlation and phenotypic correlation. Grain yield per plant shown significant positive correlation with number of pods per plant, days to flowering, days to pod setting harvest index, seed protein content and negatively significant correlation with days to maturity as well as plant height at phenotypic level. Path coefficient analysis revealed that most of the characters under study exhibited positive direct effect on grain yield per plant at both genotypic and phenotypic level. However days to pod setting shown negative direct effect on grain yield per plant yield at both genotypic and phenotypic level. Thus pods per plant, harvest index determining grain yield indicating scope for improving fieldpea yield by selection. The mutants with small pods, tall and increased number of pods per plant were isolated in M2 generation.

Hatchery production of the clownfish Amphiprion nigripes at Agatti island, Lakshadweep, India.

Healthy individuals of matured clownfish, Amphiprion nigripes and sea anemone, Heteractis magnifica were collected from the Agatti island lagoon by snorkeling. During ‘conditioning’ for 3 months, pair formation occurred and the same were transferred to rectangular fiber glass spawning tanks of 1000 l capacity. Suitable water quality parameters were maintained. The fishes were fed with tuna eggs, boiled clam meat, squid, octopus and trash fish thrice in a day. Reproductive behaviour and embryonic development were documented. Spawning took place in between 0900 - 1100 hr and hatched-out occurs, after sunset following an incubation period of 8-9 days. Size of the newly laid egg was 2.0-2.2 mm in length and 1.0-1.2 mm in width. The larval rearing tanks were maintained with UV-treated water and followed the optimal physico-chemical parameters as in the parent tanks. The different light regimes and diets were used for the successful larval rearing. The maximum larval survival (61%) was achieved at the photoperiod of 24 L/0D. Within 15-17 days, the larvae metamorphosed and took up parent colouration and comparatively high growth rate was observed when fed on algae enriched rotifer than those with poly unsaturated fatty acid (PUFA).