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??OBJECTIVE To identify seven species from Umbilicariales and study the chemical constituents, antioxidant and anti-inflammatory activities of crude products from the hot-water extraction. METHODS The seven species from Umbilicariales were identified using ITS sequence analysis, and the seven crude polysaccharides were extracted from the species by water extraction method. RESULTS Umbilicaria esculenta, Umbilicaria yunnana, Dermatocarpon miniatum and Lasallia papulosa were identified, and the main components of crude polysaccharides from the species were carbohydrates. The crude polysaccharides (JUY) from Umbilicaria yunnana of Jiangxi had the best antioxidant ability, it exhibited a higher scavenging activity of superoxide radicals (EC₅₀=0.56 mg??mL^-1) than Vc (EC₅₀=1.67 mg??mL^-1), and the scavenging activity for hydroxyl radicals (EC₅₀=0.53 mg??mL^-1) was comparable to Vc (EC₅₀=0.51 mg??mL^-1). Anti-inflammatory experimental results show that JUY has the best anti-inflammatory effects, and the better inhibitory effect of IL-1?? than dexamethasone (P<0.01). CONCLUSION The species from Umbilicariales of seven different areas were identified as four genera. The crude polysaccharides (JUY) from Umbilicaria yunnana of Jiangxi had the best antioxidant and anti-inflammatory activities.     

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??OBJECTIVE To study the preventing effects of p-coumaric acid(p-CA) on acute hypoxia-induced pulmonary edema by mice experiments. METHODS Acute-hypoxia model was established using a normobaric hypoxia chamber in vivo. Salidroside was set as a positive control drug. And the test period was 7 d using a method of intragastric administration. The measurements including pulmonary water content, HE staining, inflammatory factors, anti-oxidative indexes and Na⁺, K⁺-ATPase were performed to determine the efficacies and mechanisms of p-CA on preventive against acute hypoxia-induced pulmonary edema. RESULTS As compared with the normal group, pulmonary water contents increased significantly by 3.56% in the mice treated with acute hypoxia (9.5% O₂) for 6 h (control group) (P<0.01), and administration with p-CA (25, 100 mg??kg^-1??d^-1) for 7 d could significantly reduce this index (P<0.05), which was as effective as the positive group. The action mechanisms of p-CA could be due to its abilities of improving the activity of Na⁺, K⁺-ATPase, enhancing antioxidant capacity (SOD??, CAT?? and MDA??) and inhibiting inflammatory factors (IL-1?? and IL-6). CONCLUSION p-CA has greater preventive effects on acute hypoxia-induced pulmonary edema in mice.     

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??OBJECTIVE To investigate the anti-inflammatory effects of extract of Scutellaria baicalensis Georgi (SGE) and underlying mechanism by using LPS-induced microglial BV2 cells. METHODS MTT assay was used to observe the cell viability. The content of NO in cell supernatant was measured by Griess reagent. The levels of IL-1??, IL-6 and TNF-?? were detected by ELISA kits. The intracellular TLR4 expression was assayed by Western blotting. RESULTS The levels of NO, IL-1??, IL-6 and TNF-?? were significantly increased induced by LPS in the supernatant of BV2 cells (all P<0.01). However, co-treatment with SGE 100 ??g??mL^-1 significantly decreased the production of related inflammatory factors including NO (P<0.01), IL-1??(P<0.01), IL-6 (P<0.01) and TNF-?? (P<0.05). Furthermore, SGE significantly inhibited the TLR4 expression induced by LPS in BV2 cells. CONCLUSION SGE is able to alleviate LPS-induced inflammatory responses in BV2 cells through down-regulation of TLR4 protein expression suggesting that SGE has therapeutic potential for the treatment of neuroinflammatory diseases.     

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??OBJECTIVE To investigate the protective effect and possible mechanism of Rhizoma Coptis(RC) on lipopolysaccharide(LPS)-induced inflammatory injury in rat hepatocytes(BRL). METHODS LPS-induced BRL cells injury model was established in vitro, then the damaged cells were given different interventions and treatment with 0.175, 0.1 mg?? mL^-1 RC aqueous extract as the test drug, and dexamethasone(Dex) as positive control drug. The optimal test doses of LPS and RC aqueous extract were selected and determined by cell counting kit-8(CCK-8), the cellular apoptosis rate was determined by flow cytometry, TLR4/NF-??B and TLR4/IRF3 signaling pathways and the mRNA level of related inflammatory mediators(TNF-??, IL-1??, IL-6) were detected by RT-PCR, the NF-??B p65 protein expression was analysed by Western blot and immunofluorescence techniques. RESULTS ??Compared with normal control group, 0.1 mg??mL^-1 LPS affected on BRL cells for 24 h, the cell survival rate was decreased significantly(P<0.01), the apoptotic rate increased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 were significantly increased(P<0.01), and the NF-??B p65 protein expression was increased. ??Compared with the model group, 0.1 and 0.175 mg??mL^-1 RC affected on LPS-induced BRL cells for 24 h, the survival rate of BRL cells was increased significantly(P<0.05), the apoptotic rate decreased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 and the NF-??B p65 protein expression were decreased significantly(P<0.01). CONCLUSION Rhizoma Coptis has obviously protective effect on LPS-induced inflammatory injury in rat hepatocytes(BRL), the mechanism of which may be related with inhibiting apoptosis, reducing the release of inflammatory factors such as TNF-????IL-1?? and IL-6, blocking NF-??B p65 protein nuclear translocation, interfering the R4/NF-??B and TLR4/IRF3 signaling pathway.     

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??OBJECTIVE To investigate the in-vitro release behavior of venetoclax preparations, the pharmacokinetic processes and the correlations between in-vitro release and in-vivo absorption of venetoclax in Beagle dogs. METHODS The dissolution curves of venetoclax preparations in different dissolution media were studied. HPLC method was established for the determination of venetoclax in Beagle dogs, and the pharmacokinetics were studied for commercial venetoclax tablets in Beagle dogs under fed and fasted states.The IVIVC study was carried out by linear regression of cumulative in-vitro drug release and in-vivo absorption accumulation percentage data. RESULTS The in-vitro release behavior among venetoclax formulation in 4 dissolution media were significant difference. The simple, accurate and rapid analysis method for venetoclax blood samples was established. The fed group and the fasted group AUC_0???? were (32.38??5.87) and (27.70??6.32) mg??h??L^-1, the concentration of peak were (4.04??0.78) and (3.72 ??0.69) ??g??mL^-1, the peak time were (6.01??1.04) and (4.27??0.92) h, respectively, and there was obvious difference (P<0.05) in AUC_0???? and the peak time between two group. Percentage of in-vivo absorption was in good agreement with in-vitro release in pH 6.8 0.2%SDS media. CONCLUSION The study shows that food could improve the bioavailability of venetoclax formulation; 0.2%SDS pH 6.8 media (paddle, 75 r??min^-1) is the in-vivo release of venetoclax associated with the in-vitro release condition.     

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??OBJECTIVE To explore the effect of total coumarins isolated from Hedyotis diffusa (total coumarins from Hedyotis diffusa, TCHD) on proliferation inhibition of leukemia cells, and to explore its related mechanism. METHODS The purity of TCHD prepared by ethanol reflux extraction was tested by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) system. The cells (KG-1 Kasumi-1, THP 1 cells, U937 cells and K562 cells) were treated with TCHD(0.02, 0.04, 0.06, 0.08, 0.10 mg??mL^-1) for 24 or 48 h, the inhibitive effect of TCHD on cells growth were determined by MTT method. After Kasumi-1 cells were incubated with TCHD for 24 h,the apoptosis of cells were analyzed by flow cytometry stained with Annexin V/PI. The expression levels of caspase-3, caspase-8, caspase-9,PARP and Bcl-2 family protein were assayed by Western blot. RESULTS TCHD in certain concentration range could markedly inhibit the proliferation of AML cells, their IC₅₀ on Kasumi-1, THP-1 KG-1, U937 and K562 cells were 0.077, 0.083, 0.096, 0.087, 0.096 mg??mL^-1 for 24 h, and 0.059, 0.067, 0.072, 0.064, 0.068 mg??mL^-1 for 48 h. TCHD has significant inhibitory effect on Kasumi-1, which was stronger than those on other cell lines, and showed a dose- and time-dependent manner(r=0.357,P<0.05). The apoptotic proportion of Kasumi-1cells in 0, 0.02, 0.04, 0.06, 0.08, 0.10 mg??mL^-1 TCHD treatment groups for 24 h were (5.33??0.41)%, (7.99??0.45)%, (10.22??0.32)%, (20.10??1.99)%, (28.66??0.67)% and (33.24??2.12)%, respectively. After treated with TCHD(0.02-0.06 mg??mL^-1) for 24 h, G₀/G₁ phase ratio of Kasumi-1 detected by flow cytometry were (51.43??3.21)%, (62.91??2.35)% and (76.42??4.14)%, respectively, which were significantly higher than that of the control group (35.8??5.25)% (P<0.05).Western blot results showed that different concentrations of TCHD could activate caspase-8, caspase-9, caspase-3 and PARP, promote the expression of cyto-C, down-regulate the cyclin E and CDK6, CDK2, p-CDK2 and cyclin D₁ protein, and up-regulate the expression of p21 proteinin concentration- dependent manner(P<0.01). CONCLUSION TCHD can obviously inhibit the proliferation of Kasumi-1 in a dose- and time-dependent manner, which may relate to the apoptosis of Kasumi 1 induced by activating caspase-3, 9, PARP protein through the mitochondrial pathways and Kasumi-1 cell block in G₀/G₁ phase through the influence of CDK2, p-CDK2, CDK4/6, cyclin E, cyclin D₁ and p21.     

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??OBJECTIVE To study the influence and mechanism of bioactive ingredients of Ligusticum chuanxiong Hort. on the transport of gastrodin based on cell culture model in vitro. METHODS Cell toxicity of gastrodin, ligustilide, senkyunolide ?? and senkyunolide A were detected by MTT assay. The transport mechanism of gastrodin and the influence of ligustilide, senkyunolide ?? and senkyunolide A on the transport of gastrodin were studied in MDCK-MDR1 monolayer cells. The changed expressions of P-gp caused by ligustilide, senkyunolide ?? and senkyunolide A were analyzed by Western blotting.RESULTS Gastrodin showed relatively poor absorption in MDCK-MDR1 cells for its apparent permeability coefficients were less than 1??10^-6 cm??s^-1. P-gp inhibitor made the P_app(B??A)/P_app(A??B) of gastrodin reduced from 1.29 to 0.79, which indicated that the transport of gastrodin was influenced by P-gp. In the presence of ligustilide(30 ??g??mL^-1) or senkyunolide ??(120 ??g??mL^-1), the P_app(A??B) of gastrodin in MDCK-MDR1 were significantly increased(P<0.01). In the presence of senkyunolide A(120 ??g??mL^-1), the P_app(A??B) of gastrodin in MDCK-MDR1 were markedly increased(P<0.05). High, medium and low dose of ligustilide, senkyunolide A and senkyunolide ?? could significantly inhibit the expression of P-gp protein.CONCLUSION The results indicate that the transport mechanism of gastrodin might be passive diffusion as the dominating process with the active transportation mediated mechanism involved. Ligustilide, senkyunolide A and senkyunolide ?? increase the transport of gastrodin attribute to down-regulate P-gp expression.     

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??OBJECTIVE In those patients by monitoring and analyzing the whole blood concentrations of sirolimus of 410 cases of patients underwent kidney transplant, To provide guidance for the reasonable utilization of medications. METHODS Chemiluminescence microparticles immuno assay (CMIA) was used to determine the whole blood concentrations of sirolimus of 42 patients (410 cases) in the First Affiliated Hospital of Zhengzhou University hospital, from January 2013 to March 2015 years. Then the relationships between the whole blood concentrations of sirolimus and ALT, AST, T-CHO, TG or platelet count were analyzed. RESULTS In our study, the average whole blood concentration of sirolimus of 410 cases was (5.6??2.9) ng??mL^-1, ranging from 1.02 to 22.85 ng??mL^-1. Three hundred and six cases got concentrations of 4~8 ng??mL^-1, accounted for 74.6%. 46 cases(11.2%)got concentrations less than 4 ng??mL^-1 and 58 cases with concentrations more than 8 ng??mL^-1 accounted for 14.2%. The whole blood concentration of sirolimus of those patients showed positive correlations with ALT, AST, T-CHO, TG(P <0.05) and negative correlation with platelet count(r=-0.231, P<0.05). Patients with abnormal blood drug concentration may be related to compliance, diet, blood volume, genotype, combination and individual differences. CONCLUSION With a narrow therapeutic window, individualized dosage regimens need to be adopted and the blood concentration of which should be monitored when sirolimus was used. Close attention should be paid to the concentration of blood fat, and the blood concrentration of sirolimus need timely adjustment to guarantee a considerable efficacy and reduce the incidence of adverse reactions.     

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??OBJECTIVE To provide scientific basis for variety evaluation and selection of Dendrobium officinale. METHODS The agronomic traits of D. officinale were measured. The chemical compositions of D. officinale stem were analyzed. The dynamic changes of plant biomass and chemical composition content during mycorrhizal cultivation with M2 (Mycena sp.)were observed. RESULTS There were significant differences between D. officinale phenotype A and B in stem length, internode length, leaf width and leaf shape index (P<0.05), and in the content of polysaccharide and ethanol soluble extract (P<0.05). Moreover, the contents of 4,4??-dihydroxy-3,5-dimethoxybibenzyl and 3,4??-dihydroxy-5-methoxybibenzyl were always very different between phenotype A and B during plant growth. After 12 months of growth, the stem dry weight and plant dry weight of M-A group were significantly higher than those of CK-A group(P<0.01), increasing by 37.6% and 37.3%, respectively. The contents of polysaccharide and six marker compounds of M-A group were increased by 36.8% and 5%-50% than those in CK-A group, respectively. After 12 months of growth, the contents of polysaccharide and six marker compounds of M-B group were increased by 16.7% and 25%-170% than those in CK-B group, respectively. CONCLUSION The bibenzyl content and the response ability to mycorrhizal fungi M2 of D. officinale phenotype A and B are different.     

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??OBJECTIVE To establish a quantitative analysis of multi-components by single marker(QAMS) for the determination of three atractylenolides in Atractylodes macrocephala Koidz. METHODS UPLC method was applied to determine atractylenolide ??, ?? and ?? by external standard method (ESM)first. With this established UPLC method, atractylenolide ?? was used as the internal standard (IS)to determine the correction factors (RCFs)of the two other atractylenolides, and their contents in all the samples were calculated by their RCFs at the same time. By comparing the contents determined by the ESM and QAMS methods, the feasibility and accuracy of QAMS method were verified. RESULTS Within a certain range(atractylenolide??:3.7-59.3 ??g??mL^-1, atractylenolide ??: 3.9-63.5 ??g??mL^-1, atractylenolide ??: 7.3-116.1 ??g??mL^-1), the RCFs of atractylenolide ??, ?? to atractylenolide ?? were 0.633 and 1.895, respectively, which had good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method(P??0.05). CONCLUSION The QAMS method is feasible and accurate for the determination of the three atractylenolides and is beneficial to the quality control of Atractylodes macrocephala Koidz..