OBJECTIVE To investigate the role of cinnabar and realgar in Angong Niuhuang Wan (AGNH) -produced neuroprotection against lipopolysaccharide ( LPS) -mediated neuronal damage and further explore the corresponding mechanisms. METHODS Primary rat midbrain neuron-glia cultures were used as an in vitro model to investigate effects of AGNH on LPS-mediated degeneration of dopamine (DA) neurons. The experiment was divided into normal control group, LPS model group, LPS + cinnabar (4 and 40 mg·L-1) groups, LPS + realgar (4 and 40 mg·L-1 ) groups and LPS + AGNH (40 and 400 mg·L-1 ) group. Drugs were added 30 min before LPS treatment. After 7 d, dopaminergic neurotoxicity was assessed through the quantification of tyrosine hydroxylase (TH)-positive neurons and morphological analysis of TH-positive neurons; the activation of microglia was evaluated using OX-42 antibody; the gene expression of tumor necrosis factor-α (TNF-α) and induced nitric oxide synthase (iNOS) mRNA in microglia was performed by real-time RT-PCR analysis, and the release of TNF-α and nitric oxide (NO) in the supernatant of neuron-glia cultures was determined respectively by the ELISA and Griess reagent. RESULTS Compared with normal control group, DA neurons in LPS model group decreased by 40% (P <0.05) , microglial activation was induced, the expression of TNF-α mRNA and iNOS mRNA in microglia increased 9 and 2 times, respectively ( P < 0. 05 ) , and subsequent production of TNF-α and NO in the supernatant of neuron-glia cultures increased 20 and 30 times, respectively (P<0.05). Compared with LPS model group, AGNH 400 mg·L-1 and realgar 40 mg·L-1 significantly attenuated LPS-mediated DA neuronal loss by 40% and 30% , respectively (P<0.05) and inhibited activation of microglia and expression of TNF-α mRNA by 61% and 52% (P <0.05). iNOS mRNA was reduced by 58% and 51% (P <0.05 ) in microglia. The subsequent release of TNF-α was reduced by 55% and 43% (P<0.05) and NO reduced by 53% and 34% (P<0.05) in the supernatant of neuron-glia cultures. Cinnabar had no inhibitory effect on LPS-induced changes. CONCLUSION AGNH protects LPS-induced neurotoxicity through its anti-inflammatory properties and realgar might be the key contributor to the neuroprotective action of AGNH, while cinnabar fails to show any neuroprotection.

Objective To evaluate the effects of epidural infusion of different concentrations of lidocaine on the amount of propofol needed to induce anesthesia and the end-tidal isoflurane concentration needed to maintain BIS at 50-55 during operation. Methods Forty-five ASA Ⅰ-Ⅱ patients of both sexes (24 males, 21 females) aged 21-60 yrs, weighing 46-77 kg undergoing elective upper abdominal surgery were randomly divided into 3 groups with 15 patients in each group : group Ⅰ epidural normal saline ( NS); group Ⅱ epidural 1 % lidocaine (Lido) and group Ⅲ 2 % Lido. An epidural catheter was inserted at T9-10 into epidural space and advanced 3-4 cm cephalad. After a test dose of 3 ml, NS or 1% or 2% Lido 10 ml was injected into the epidural space. Anesthesia was induced with propofol infusion at 25 mg?min-1. Propofol infusion was stopped when the patients stopped responding to loud voice (OAAS/2) and the amount of propofol infused was recorded. Then fentanyl 2 ?g?kg-1 and vecuronium 0.15 mg?kg-1 were given i.v. . The patients were intubated and mechanically ventilated and PETC02 was maintained at 35 mm Hg. Anesthesia was maintained with isoflurane inhalation and epidural infusion of Lido or NS at 7 ml?h-1 . BIS was maintained at 50-55 during operation. Results The amount of propofol needed to induce anesthesia was 1.23?0.34 mg?kg-1 in group Ⅱ and 1.02?0.25 mg?kg-1 in group Ⅲ compared to 1.67?0.38 mg?kg-1 in control group (group Ⅰ) (P

Objective To investigate optimal extraction process of Piper puberulum ( Benth.) Maxim.and qualitative analyze the chemical component of the extracts.Methods Method of solvent heating reflux was used for extraction.On the basis of single factor experiment, L9 (34 ) orthogonal experiment was designed with the variants of extraction frequency, time, material-liquid ratio, and immersion time.Extraction rate as index, extraction processes were optimized to achieve best extraction.The extracts, including total extract, water elution, and ethanol elution, were physiochemically analysed to achieve an initial qualitative result.Results The optimal extraction process was: extractions 3 times for 2 hours, with an 1︰30 material -liquid ratio and 2 hours of immersion, Initial qualitative analyzed the total extracts containing amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, saponins, polysaccharides, reducing sugars or glucosides, cumarins, terpene lactones, phenols, and tannins.The water elution containing: amino acids, polypeptides, proteins, saponins, polysaccharides, reducing sugars or glucosides, cumarins, and terpene lactones.The ethanol elution containing: amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, polysaccharides, reducing sugars or glucosides, phenols, and tanins.Conclusion The experiments show that optimal extraction process can achieve high extraction yield, stable and practical.

Objective To investigate the effects of tetrahydroxystilbene glucoside (TSG) on cytochrome P450(CYP) in mouse livers .Methods Kunming male mice were divided into the blank ,low dose and the high dose of TSG groups .3 ,5 and 7 after intra-gastrical administration of TSG ,mice were sacrificed and the mRNA expressions of CYP isoenzymes in mouse livers were measured by real time reverse transcription-polymerase chain reaction(RT-PCR) ,respectively .Results TSG significantly inhibited CYP1A2 and CYP 3A4 mRNA expressions at 3th ,5th and 7th day after treatment .TSG time-dependently increased CYP2E1 mRNA expres-sion .TSG inhibited CYP4A14 mRNA expression at 7th day after treatment .Moreover ,TSG had no significant effects on CYP2B10 , CYP3A11 and CYP3A25 mRNA expressions .Conclusion TSG has significant effects on CYP1A2 ,CYP2E1 ,CYP3A4 and CYP4A14 mRNA expressions but no significant effects on CYP2B10 ,CYP3A11 and CYP3A25 mRNA expressions .

AIM Taurine was reported neuroprotective under several ischemic models in vivo. In this study, the direct effect of taurine against oxygen-glucose deprivation (OGD) inducing acute neuronal injury and the underlying mechanisms in vitro were investigated. METHODSFour hours OGD was used to induce in vitro ischemic injury in rat cortical neurons. Taurine 5, 10 and 20 mmol·L-1 was added 20 h before and during 4 h OGD period respectively. Mortality rate of neuron was assayed by MTT and flow cytometry methods. Level of neuronal [Ca2+]i was detected by Fura 2/AM loading. Amino acid concentrations in culture media were measured by high performance liquid chromatography. RESULTS Under OGD conditions, neuronal death was markedly increased, and the levels of neuronal [Ca2+]i and extracellular glutamate level were enhanced obviously. Taurine pretreatment obviously decreased the percentage of neuronal death induced by OGD. In addition, abnormal elevation of neuronal [Ca2+]i and extracellular glutamate level induced by OGD both were markedly repressed by taurine. CONCLUSION Taurine can alleviate rat cortical neuron injury induced by OGD, the mechanisms were likely due to repressing calcium overload and inhibiting excessive release or leakage of glutamate under such conditions.

Aim To investigate the protective effect of noble dendrobium polysaccharides ( NDP ) on lipopo-lysaccharide ( LPS)-induced neuron injuries in newborn rat cerebral cortex glial cells and neuron mixed cul-tures.Methods The primary cultures of newborn rat cortical neurons and glial cells were established and the existence of the neurons , astrocytes and microglia was verified respectively .NDP was given to LPS-induced mixed cultures , the mRNA levels of IL-1β, TNF-αand COX-2 were assayed by real time PCR .Results NDP reduced the glial cell activation and neuron dam-age after it was given to LPS-induced mixed cultures . The mRNA levels of IL-1β, TNF-α, COX-2 were re-duced .Conclusion NDP protects against LPS-in-duced neuron-inflammation in neurons and glial cells cultures.

Aim To investigate the effect and mechanism of cobra venom metalloproteinase atrase A on inhibiting platelet aggregation.Methods Platelet aggregation induced by collagen,ADP,PAF,AA,ristocetin and thrombin,respectively,was measured turbimetrically after platelet incubated with atrase A.Western blot was used to detect the effect of atrase A on cleavage of platelet membrane glycoprotein and von Willebrand Factor.Results Atrase A significantly inhibited platelet aggregation induced by ristocetin and thrombin in a dose-and time-dependent manner.Meanwhile,atrase A just showed slight inhibitory effect on platelet aggregation induced by PAF,AA,collagen,and ADP after incubated with PRP for 5 min.After incubation time was prolonged to 30 min,significant inhibition was shown on platelet aggregation.Western blot revealed that atrase A cleaved platelet membrane glycoprotein GPIb.Conclusions Cobra venom metalloproteinase atrase A significantly inhibits platelet aggregation induced by ristocetin and thrombin due to the cleavage of platelet membrane glycoprotein Ib.And atrase A also has inhibitory effect on platelet aggregation induced by ADP,PAF,AA,and collagen.

Aim To investigate the inhibitory effect of atrase B on human platelet aggregation induced by activated complement.Methods By employing CVF to activate complement,the effect of atrase B on gel filtered platelet aggregation induced by activated complement was measured by turbidimetry and the expression of P-selectin and GPⅡb/Ⅲa on platelet membrane were detected by flow cytometry.Results Atrase B inhibited platelet aggregation and the expression of P-selectin and GPⅡb/Ⅲa on platelet membrane induced by activated complement.Conclusion Anticomplementary protein atrase B from Naja atra venom can significantly inhibit platelet activation and aggregation induced by activated complement.

Objective To study the effect of protopine (Pro) on the intracellular free calcium ions of the smooth muscular cells of the thoracic aorta in rats. Methods The procedure of calcium absence-calcium addition was designed to observe the changes of the level of calcium ions in the strips of the smooth muscle of the thoracic aorta indirectly. The concentration of calcium ions in the smooth muscle of the thoracic aorta was determined with Fura-2/AM loaded SMCs. The elevation of calcium ions was induced with norepinephrine (NE). The concentration of potassium ions was observed in the presence of Pro. Results Pro significantly inhibited the NE-induced transient contract of the smooth muscle of the thoracic aorta in calcium-free medium and long-lasting contraction after the addition of calcium ions in a concentration-dependent manner. In Fura-2/AM loaded SMCs, Pro (50 ?mol/L or 100 ?mol/L) exerted no effect on the resting calcium ions but obvious effect on the NE-induced and high potassium level-induced elevation of calcium ions. In the presence of extracellular calcium ions, Pro (50 ?mol/L) decreased the NE-induced elevation of calcium ions but showed no effect on potassium-induced elevation of calcium. Pro (100 ?mol/L) significantly decreased NE-induced and high potassium level-induced elevation of calcium ions. Conclusion Pro reduces NE-induced or high potassium level-induced elevation of calcium ions in the smooth muscle of the thoracic aorta through its effect on calcium release and/or calcium influx.

OBJECTIVE To observe the protective effect and underlying mechanism of total ginsenosides (TG) on bleomycin-induced pulmonary fibrosis. METHODS Intratracheal instillation of bleomycin 5 mg · kg-1 was conducted to establish a pulmonary fibrosis mouse model. Kunming mice(1/2 males and 1/2 females)were randomly divided into sham-operation(Sham),model,total ginsen?osides 40,80 and 160 mg·kg-1 and prednisone acetate(5 mg·kg-1) groups. After 28 d administration,the histopathological changes in the lung were analyzed by hematoxylin eosin(HE)and Masson staining. The exprssion of alpha smooth muscle actin(α-SMA)in the lung was detected by real-time PCR. The content of hydroxyproline(HYP)and glutathione(GSH),level of total antioxidant capacity(T-AOC)and hydroxy radical(·OH),activity of myeloperoxidase(MPO)and nitric oxide synthase(NOS)in the lung were detected by corresponding kits. RESULTS Compared with the sham group,the pulmonary indexes in model group were significantly increased(P<0.01),alveolitis and pulmonary fibrosis were obvious. The mRNA expression ofα-SMA,content of HYP and · OH,activity of MPO and NOS were increased(P<0.05),but the content of GSH and T-AOC in model group was decreased(P<0.05). Compared with model group,the pulmonary indexes in TG 80 and 160 mg · kg-1 and prednisone acetate 5 mg · kg-1 groups were reduced(P<0.05),and the degree of alveolitis and pulmonary fibrosis was mitigated. The mRNA expression ofα-SMA,content of HYP and · OH, the activity of MPO and NO were decreased (P<0.05),while the content of GSH and T-AOC was increased(P<0.05). CONCLUSION TG can improve the degree of mice pulmonary fibrosis induced by bleomycin. The mechanism may be related to the increased antioxidant capacity of organisms.