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OBJECTIVE： To study the chemical constituents of supercritical CO2 extract from Xinjiang Daucus carota seed（called “D. carota seed extract” for short）， and to preliminarily evaluate its antimicrobial and antioxident activity， so as to provide reference for the development and quality control of related products of the plant. METHODS： The non-polar volatile part of Xinjiang D. carota seed was extracted by supercritical CO2 extraction method. The chemical constituents of the extract were identified and quantified by GC-MS and standard spectral labrary. The relative percentage content of each component was calculated by peak area normalization method. The antibacterial activity of D. carota seed extract against Staphylococcus aureus，Escherichia coli and Candida albicans were investigated by trace broth dilution method and AGAR medium plate method. Its antioxidant activity was investigated by TLC-bioautography method. RESULTS： Twenty chemical constituents were identified from D. carota seed extract （the total relative percentage content was 98.76％）， mainly including β-bisabolene， α-asarone， α-pinene， β-caryophyllene， aspidinol， etc. D. carota seed extract has certain antimicrobial activity on S. aureus， E. coli and C. albicans. The minimum inhibitory concentrations were 0.039， 0.833， 0.625 mg/mL， and minimum bactericidal concentrations were 0.078， 1.667， 1.250 mg/mL， respectively. The extract has a certain scavenging ability to DPPH radical. CONCLUSIONS： D. carota seed extract obtained by supercritical CO2 extraction contains monoterpenes and sesquiterpenes， and possesses certain antibacterial and antioxidant activity.
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Objective Find abnormal changes of plasma lipid metabolism-related proteins before 20 weeks of gestation in patients with hypertensive disorder of pregnancy(HDP), and preliminarily investigate the role of plasma apolipoprotein C4 elevation in HDP. Methods A nested case-control study was used. The plasma were collected from pregnant women who underwent routine prenatal examination in Guangzhou Women and Children's Medical Center from November 2014 to March 2017. Label-free mass spectrometry was used to detect the differences in plasma lipid metabolism-related proteins before 20 weeks of gestation between 12 pairs of HDP patients and normal controls, and different 48 pairs of samples were used for verification. The protein with the most significant difference multiples was screened to study its effects on monolayer permeability and nitric oxide secretion of endothelial cells. One-way ANOVA was used for comparison between groups, and P<0.05 was considered as statistically significant difference. Results Compared with the control, the lipid metabolism-related proteins, APOC4, Fatty acid-binding protein 4 (FABP4), Apolipoprotein E (APOE), Apolipoprotein C3 (APOC3) and Beta-2-glycoprotein 1(APOH) raised to 1.94, 1.82, 1.59, 1.55 and 1.38 times, phospholipid transfer protein (PLTP) decreased to 0.78 times in plasma before 20 weeks of pregnancy of patients with HDP (t value were 2.499, 2.497, 2.081, 2.098, 2.426 and 2.564, respectively, P<0.05). Cell experiments results showed that 50 ng / ml APOC4 significantly increased 20％ HUVEC single layer cell permeability to FITC-labeled dextran (F=455.4, P<0.01), and significantly decreased the level of nitric oxide in the supernatant of HUVEC culture by 25％ (F=61.92, P<0.01). Conclusions Before diagnosis, plasma protein levels involved in lipid metabolism in HDP patients have been changed, resulting in abnormal lipid metabolism. APOC4 can increase the permeability of vascular endothelial cells, inhibit endothelial source of NO secretion, cause endothelial dysfunction.
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Objective Newborn bilirubin encephalopathy seriously threatens the life and health of newborns, Both the mortality and morbidity are high.Cerebrospinal fluid bilirubin concentration can better reflect the blood-brain barrier function and brain bilirubin levels, which can help the diagnosis of bilirubin encephalopathy.Methods In order to meet the clinical needs, we confirmed the detection performance of the existing serum bilirubin detection system for cerebrospinal fluid bilirubin, including precision, accuracy and analysis of measurement range.Results The results showed that the detection system was linear at 1.0-25.1μmol/L, and the daytime CVat normal and pathological values was less than the precision requirement (6％) .The bias of each sample and the average bias are less than the allowable bias (5％) , the total error is less than the total allowable error (15％) .Conclusion Therefore, the performance of the detection system meet the industry standards, can be used for neonatal cerebrospinal fluid bilirubin detection.
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<p><b>OBJECTIVE</b>To establish a highly sensitive screening method for phytoestrogen active constituents and to primarily screen the phytoestrogenic active constituents from the chickpea extractions by the method.</p><p><b>METHOD</b>Human ERalpha cDNA was cloned using MCF-7 total RNA as the template by RT-PCR and then was constructed into a pcDNA3 and named as pERalpha. The cell line MCF-7 was co-transfected with pERalpha and the reporter plasmid pERE-Luc which carrying the estrogen response element (ERE) plus the luciferase reporter gene. The luciferase activity was then assayed. The model was optimized by changing the ratio of two plasmids. The feasibility of the optimized model was further proved by the several known phytoestrogen compounds including fermononetin, biochanin A and genistein, et al. As an application of the model, the phytoestrogen activity of the extracts of the chickpea was assayed.</p><p><b>RESULT</b>The recombinant plasmid (pERalpha) can enhance luciferase activities of pERE-Luc transfected MCF-7 cells. The highest transfection efficiency and luciferase activity were found at the ratio of 10:1 (pERE-Luc: pERalpha), the luciferase activity was improved five times as high as the unique pERE-Luc transfection. The co-transfection screening model also indicated that fermononetin, biochanin A and genistein could induce ERE-driven luciferase activity and ICI 182,780 suppressed the induced transcription. As the application of the model, the results showed that the ethanol (70%) total extraction, the ethyl acetate extraction and the ligarine extraction of the chickpea can induce ERE-driven luciferase activity. Concurrent treatment with ICI 182,780 abolished the induced luciferase activity.</p><p><b>CONCLUSION</b>A phytoestrogen active constituent screening mode have been established based on co-transfection method. It is sensitive to assay the phytoestrogen active constituents and can be applied to screen the active component of phytoestrogens.</p>