OBJECTIVE:To establish the RP-HPLC method for simultaneous determination of flavonoids(quercetin,cyanidenon,spinacetin,and 6-methoxyl-cyanidenon) in Inula britannica. METHODS:The samples were separated on chromatographic column Inertsil ODS-3(250 mm?4.6 mm,5 ?m) at a column temperature of 40 ℃.The mobile phase consisted of methanol-acetonitrile-0.49% citric acid(gradient elution) at a flow rate of 1.0 min?mL-1. The detection wavelength was set at 360 nm. RESULTS:The linear ranges of quercetin,cyanidenon,spinacetin,and 6-methoxyl-cyanidenon were 0.17～6.8 ?g(r=0.999 9),0.088～3.52 ?g(r=0.999 9),0.020 2～0.808 ?g(r=0.999 9) and 0.045～1.8 ?g(r=0.999 6),respectively,and the average recoveries of all stood at 96.03%～99.78% and RSD at 0.73%～1.36%(n=6 each). CONCLUSION:The method is simple,sensitive,accurate and reproducible,and it is applicable for the quality control of the raw medicinal substance and preparation of I.britannica.

OBJECTIVE:To study the flavonoids in Inula britannica.METHODS:The Inula britannica was extracted with 95% ethanol and isolated and purified by different column chromatography,the structures of the constituents were derived by measuring the physicochemical constants and spectral analysis.RESULTS:A total of 10 flavonoids were isolated from Inula britannica which were stated as follows:Spinacetin(1),6-methoxy-luteolin(2),Iuteolin(3),Quercetin(4),Isorhamnetin(5),Kaempferol(6),Rutin(7),Patulitrin(8),Isoquercitrin(9)and Quercitrin(10).Of which,compounds 5,7 and 10 were isolated from Inula britannica for the first time.CONCLUSION: This result serves as a scientific basis for the further study of Inula britannica.

Objective To investigate the location of CYLD in the neurons and explore the expression of CYLD in OGD/reperfusion-induced neuronal necroptosis.Methods Primary cortical neurons were cultured for 6days and neuronal purity was observed by double staining immunofluorescence of β3-tubulin and DAPI.The location of CYLD was identified by double staining immunofluorescence of NeuN,DAPI and CYLD using primary cortical neurons cultured for 14 days.Then,primary cortical neurons were divided into 8 groups:Control,EBSS,DMSO,OGD/reperfusion(0 h,2 h,6 h,8 h,12 h).Neurons were pretreated with zVAD-fmk for 30 min,OGD for 2 h and the levels of CYLD were evaluated after reoxygenation at different time points.The peak value(8 h) was chosen as reoxygenation time point.Neurons were divided into two groups as Control and OGD.The levels of CYLD were determined in both cytoplasm and nucleus after OGD 2 h and reoxygenation 8 h.Results The double staining immunofluorescence showed that neuronal cultured purity was about 70％ and the CYLD strongly expressed in nucleus but weakly in cytoplasm.The levels of CYLD increased gradually with different reoxygenation time and arrived at peak value after reoxygenation for 8 h (P ＜ 0.05),which was in accordance with the change of LDH (P ＜0.05) (Control (1.00±0.00),EBSS (1.07 ±0.03),DMSO (1.09 ±0.03),0h (1.40±0.12),2 h (1.74±0.08),6 h (2.25 ± 0.12),8 h (2.97 ± 0.15),12 h (3.01 ± 0.08)).The level of cytoplasm CYLD increased significantly in the OGD group (reoxygenation for 8 h)than that in control group (P＜0.05).But the level of nucleus CYLD had no difference between OGD and control group (P ＞ 0.05),which was in accordance with the results of immunofluorescence.Conclusion The CYLD in neurons cytoplasm is involved in necroptosis induced by OGD/deprivation and downregulating of CYLD has a protective effect on the brain injury resulted from ischemia/ reperfusion.

BACKGROUND: The transcription factor Runx2 is the key factor that regulates osteogenic differention and bone development. It has been reported that the C2C12 mesenchymal cells can be induced to differentiate into osteoblasts by Runx2 overexpression, but the molecular mechanism of induction is stil largely unclear. OBJECTIVE: To investigate the role of the members of the miR-376 family during Runx2-induced osteogenic differentiation in C2C12 cells. METHODS: The expression of the members of the miR-376 family was detected by real-time quantitative PCR at different time points using C2C12/Runx2Dox sub-line with conditional Runx2 expression. In miR-376b-3p-transfected C2C12/Runx2Dox cells, the expression of osteoblast markers, such as alkaline phosphatase and osteocalcin, was detected by real-time quantitative PCR, and the alkaline phosphatase activity was also examined by alkaline phosphatase staining. The putative miR-376b-3p targets were commonly predicted by online tools (miRanda, miRWalk and TargetScan). The functional classification of these putative targets was performed by DAVID Bioinformatics Resources database. RESULTS AND CONCLUSION: The expression of miR-376b-3p was significantly increased during Runx2-induced osteogenic differentiation of C2C12 cells, but the expression of other members was not changed. Transfection of miR-376b-3p mimic upregulated alkaline phosphatase expression, but had no effect on osteocalcin expression. The alkaline phosphatase activity was also increased by transfection of miR-376b-3p. The functional classification of miR-376b-3p putative targets showed that miR-376b-3p is involved in the skeleton development, indicating the role of miR-376b-3p in osteoblast differentiation. Taken together, these results suggest that Runx2 promotes early osteogenic differentiation in C2C12 cells by regulating the expression of genes related to osteogenic differentiation through upregulation of miR-376b-3p.

Objective To explore the effect of L-dopa on plasma homocysteine and folic acid in patients with parkinsons' s disease(PD).Methods Twenty eight elderly PD patients and thirty normal subjects were enrolled in this group.The homocysteine,cobalamin and folate were examined in normal group and in PD group before treatment and after being treated with L-dopa for six moths respectively.Then the homocysteine,cobalamin and folate were compared between the two groups.Results The plasma homocysteine levels increased in PD patient group after being treated with L-dopa for six months(19.19? 8.01)?mol/L as compared with those of the PD group before treatment(12.50?3.78)?mol/L and those of control group(12.60?3.94)?mol/L(P

Objective To evaluate the effects of propofol and sevoflurane anesthesia on cognitive function and amyloid beta protein ( Aβ) deposition in hippocampi of aged mice. Methods Thirty?six SAMP8 mice, aged 6 months, weighing 29-32 g, were randomly assigned into 4 groups ( n=9 each) using a random number table: control group ( group C ) , propofol anesthesia group ( group P ) , sevoflurane anesthesia group (group S) and propofol plus sevoflurane anesthesia group (group PS). In group P, propofol 140 mg∕kg was injected intraperitoneally, when righting reflex occurred, additional propofol 70 mg∕kg was given, and when it occurred again, additional propofol 40 mg∕kg was given. Group S continuously inhaled 1% sevoflurane for 120 min. Group PS continuously inhaled 2% sevoflurane for 120 min, and when righting reflex occurred, additional propofol 40 mg∕kg was given. Anesthesia was maintained for 120 min in P, S and PS groups. Before anesthesia and at 7, 14 and 28 days after anesthesia, Morris water maze test was performed, and the escape latency was recorded. Hippocampi were obtained to determine the expression of Aβ using immuno?histochemistry. Results Compared with group C, the escape latency was significantly prolonged at 7 days after anesthesia, and the expression of Aβwas up?regulated at 7, 14 and 28 days after anesthesia in group S, and no significant change was found in the parameters mentioned above in P and PS groups. Compared with the value at 7 days after anesthesia, the expression of Aβ was significantly down?regulated at 14 and 28 days after anesthesia in group S, and no significant change was found in the expression of Aβ at 14 and 28 days after anesthesia in C, P and PS groups. Conclusion Although sevoflurane anesthesia promotes Aβ deposition in hippocampi, it only causes short?term cognitive dysfunction, however, anesthesia with propofol or with propofol in combination with sevoflurane produces no influence in aged mice.

Objective:To establish a quality standard for vinegar frankincense in Liuwei Jingkang capsules. Methods: TLC was used for the identification of vinegar frankincense. HPLC was used for the content determination of acetyl-11-keto-β-boswellic acid (AKBA), which was the main active component in vinegar frankincense. A SHIMADZU Shim-pack VP-ODS(250 mm × 4. 6 mm,5μm) column was used. The mobile phase consisted of acetonitrile and water containing 0. 01 mol·L-1 hydrochloric acid (78 ∶22) at a flow rate of 1. 5 ml·min-1 . The column temperature was 30℃. The detection wavelength was 252 nm, and the injection volume was 10 μl. Results:The TLC method could identify the characteristic fluorescence of vinegar frankincense was without interference from the blank. There was a good linear relationship of AKBA within the concentration range of 0. 036 5-0. 730 8 mg·ml-1(r=0. 999 7). The average recovery was 98. 24% (RSD=0. 83%, n=9). Conclusion:The established method is accurate, highly sensitive and well re-producible, which can be used for the quality control of Liuwei Jingkang capsules.

Objective To investigate whether antihypertensive treatment is beneficial to patients with diabetes mellitus when their blood pressure (BP) is below 140/90 mm Hg(1 mm Hg =0.133 kPa).Methods MEDLINE,EMBASE,IPA database and secondary resources were searched with terms including blood pressure,hypertension and anti-hypertension drug.Inclusion criteria:random control study; subjects were patients with diabetes mellitus or impaired glucose tolerance; endpoint BP ≤ 140/90 mm Hg; endpoint BP between two groups had significant differences.There were 16 studies meet inclusive criteria with a total of 51 470 patients.RR and 95％ CI were used as index to judge the difference of clinical outcomes between aggressive antihypertensive treatment group and standard antihypertensive treatment group.RevMan5.0 software was used for statistical analysis.Results When BP of patients with diabetes mellitus were below 140/90 mm Hg,anti-hypertensive treatment could reduce incidence rate of cardiovascular event (RR 0.91,95％ CI 0.87-0.96,P =0.0004) and stroke (RR 0.75,95 ％ CI 0.63-0.88,P =0.0005),and increased incidence rate of symptomatic hypotension (RR 3.57,95％ CI 1.41-11.20,P =0.03) and hyperpotassemia (RR 1.57,95％ CI 1.05-2.33,P =0.03).There were no significant differences in all-cause mortality (RR 0.94,95％ CI 0.87-1.01,P =0.08),cardiovascular mortality (RR 0.95,95％ CI 0.85-1.08,P =0.05),myocardial infarction (RR 0.93,95％ CI 0.82-1.05,P =0.26),heart failure (RR 0.90,95％ CI 0.76-1.06,P =0.21) between the aggressive antihypertensive treatment group and standard antihypertensive treatment group.Conclusions When blood pressure of patients with diabetes mellitus was below 140 mm Hg,there was little benefit from aggressive antihypertensive treatment,and the risk of serious adverse events even increased.

Objective To investigate the relationship between the oxidized low-density lipoprotein(ox-LDL) in the blood from the coronary sinus and impaired endothelium-dependent vasodilation in patients with coronary artery disease(CHD).Methods Four groups of patients were subjected,including acute myocardial infarction group(AMI,n=22),unstable angina group(UA,n=29),and stable angina group(SA, n=25) and control group(n=20) who was first suspected as CHD and then verified with normal coronary angiograms.Blood form the coronary sinus was collected through cardiac cathetering and the ox-LDL level was measured by Sandwich ELISA method.Brachial arterial hyperemia-induced flow mediated dilation(FMD) and sublingual nitroglycerin(NTG) mediated vasodilation were measured by high resolution ultrasound.Results The blood level of ox-LDL in patients with CHD was markedly higher than those in control group(P

Humans ; Multiple Myeloma ; blood ; diagnosis ; Plasma Cells ; cytology ; Plasmacytoma ; blood ; diagnosis