Objective To investigate the expression of netrin-1 in placenta from patients with pre- eclampsia and the relation to placental angiogenesis.Methods Twenty patients with pre-eclampsia(12 mild cases and 8 severe cases)and 20 normal late pregnant women were investigated.The expression of netrin-1 mRNA and protein was determined with RT-PCR and Western blotting respectively.The placenta vascular density was examined by immunohistochemical F8 staining.Results (1)The values of netrin-1 mRNA in normal group and pre-eclampsia groups were 0.51?0.08 and 0.41?0.06;The values of netrin- 1 protein in normal group and pre-eclampsia groups were 26.4?1.8 and 20.5?1.3(P

Congresses as Topic ; Gastrointestinal Diseases ; physiopathology ; therapy ; Humans ; Integrative Medicine

OBJECTIVE:To observe therapeutic efficacy and safety of Kouyanqing granules for chronic periodontitis and it effects on inflammatory cytokine. METHODS:110 patients with chronic periodontitis were divided into control group(55 cases)and observa-tion group(55 cases). Control group accepted the periodontal basic treatment as oral hygiene instruction,supragingival scaling,subgin-gival scaling and root planning. Observation group was additionally given Kouyanqing granules 20 g orally,twice a day,on the basis of control group. Both groups were treated for 4 weeks. Clinical efficacies of 2 groups were observed as well as the levels of plaque in-dex(PLI),gingival index(GI),probing depth(PD),clinical attachment level(CAL),IL-17,IL-21,and the occurrence of ADR. RE-SULTS:Both groups completed the treatment,and no patient lost to follow up. Total response rate of observation group was significant-ly higher than that of control group,with statistical significance(P<0.05). After treatment,the levels of PLI,GI,PD,CAL,IL-17 and IL-21 in 2 groups were significantly lower than before,and the observation group was significantly lower than the control group, with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Based on routine treatment,Kouyanqing granules show significant therapeutic efficacy for chronic periodontitis, can significantly improve periodontal symptom and reduce inflammatory factor level,but do not increase the incidence of ADR.

The fermentation conditions of alkaline lipase producing by Pseudomonas cepacia PCL-3 were optimized.Based on the analysis of single factorial experiments,dextrin was the most suitable carbon source,peptone and urea were the suitable compound nitrogen sources among the examined materials.Three significant factors(urea,inoculum and initial pH) were selected from the eight factors related to lipase production by Plaekett-Burman method,and were further optimized with response surface analysis.And then,steepest ascent procedures were applied to define the optimal response region of the three factors.The obtained optimal conditions were urea 0.15%,inoculum 3.05% and initial pH 8.38,under which conditions,the enzyme activity was improved from 25.37 U/ml to 48.88 U/ml,enhanced 1.93 folds.Starting from the flask conditions,the highest lipase activity of 47.69U/ml was achieved by batch fermentation in a 10 L fermentor after 52 h of the cultivation.

A solution with different Cu supply levels was cultured to investigate gama-aminobutyric acid (GABA) accumulation in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species. Increasing Cu from 0.25 to 500 micromol/L significantly enhanced levels of GABA and histidine (His), but considerably decreased levels of aspartate (Asp) and glutamate (Glu) in the leaves. The leaf Asp level negatively correlated with leaf Cu level, while leaf GABA level positively correlated with leaf Cu level. The leaf Glu level negatively correlated with leaf GABA level in Elsholtzia splendens. The depletion of leaf Glu may be related to the enhanced synthesis of leaf GABA under Cu stress.
Copper ; toxicity ; Dose-Response Relationship, Drug ; Drug Tolerance ; physiology ; Gene Expression Regulation ; drug effects ; Lamiaceae ; drug effects ; metabolism ; Plant Leaves ; drug effects ; metabolism ; gamma-Aminobutyric Acid ; metabolism

The fermentation conditions of lipase production by Geotrichum candidum Y162 were optimized. Initially, the most suitable carbon olive oil, nitrogen source soybean flour and NH4Cl, salt BaCl2 and MgCl2 were selected according to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and three statistically significant factors olive oil, BaCl2 and NH4Cl were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows: olive oil 2.35%, BaCl2 0.36%,and NH4Cl 4.69%.The optimization of culture conditions of G.candidum Y162 led to a 2.25-fold increase in lipase production relative to initial result 14.16 U/ml, which indicate that single factor in combination with response surface methodology is an effective method for optimization of lipase production conditions by G.candidum Y162.

Bioimprinting is a new developed technique to improve the characteristics of enzymes.Bioimprinting by lauric acid was conducted to improve the esterification activity of lipase PS in sol-gel immobilization process with methyltrimethoxysila(MTMS) and tetramethoxysila(TMOS) as the precursors.Results generated by checking the esterification activity and scanning electron microscope showed that bioimprinting can enhance the specific activity and thermal stability of lipase PS.The bioimprinting system was optimized by orthogonal experiment,and the optimal condition for lipase bioimprinting is water/silane molar ration(R) 12,polyethylene glycol(PEG) 120?l,and lauric acid 0.15 mmol.Compared with the free enzyme and the non-imprinted enzymes,the specific activity of imprinted enzymes has been improved 44.3 fold and 2.4 fold,respectively.Imprinted lipase show better thermal stability,and the relative activity is 58% after incubated in 80 ℃ for 0.5 h,while no activity was detected for the free enzyme.

From the N-terminal amino acid sequence of the Aspergillus niger F044 lipase,a potential homologous gene A84689 to the anl(the gene encoding the Aspergillus niger lipase)was found by means of bioinformatics.Based on the nucleotide sequence of the A84689,primers were designed to amplify anl.Nucleotide sequencing of the genomic anl gene revealed an open reading frame of 1 044 nucleotides,containing three introns(54,45 and 51 nucleotides).The deduced amino acid sequence of the anl gene corresponds to 297 amino acid residues including a signal sequence of 27 amino acid residues.The cloned cDNA coding for mature Anl(the protein of the Aspergillus niger lipase)was overexpressed in Escherichia coli BL21(De3),and the recombinant Anl was purified.The denatured recombinant Anl by 8mol/L urea was refolded in vitro by dilution and DEAE Sepharose Fast Flow chromatography.

A solution with different Cu supply levels was cultured to investigate gama-aminobutyric acid (GABA) accumulation in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species. Increasing Cu from 0.25 to 500 ?mol/L significantly enhanced levels of GABA and histidine (His), but considerably decreased levels of aspartate (Asp) and glutamate (Glu) in the leaves. The leaf Asp level negatively correlated with leaf Cu level, while leaf GABA level positively correlated with leaf Cu level. The leaf Glu level negatively correlated with leaf GABA level in Elsholtzia splendens. The depletion of leaf Glu may be related to the enhanced synthesis ofleafGABA under Cu stress.

Copper accumulation and intracellular distribution in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species, was investigated by transmission electron microscope (TEM) and gradient centrifugation techniques.Copper concentrations in roots, stems and leaves of E. splendens increased with increasing Cu levels in solution. After exposure to 500 μmol/L Cu for 8 d, about 1000 mg/kg Cu were accumulated in the stem and 250 mg/kg Cu in the leaf of E. splendens. At 50μmol/L Cu, no significant toxicity was observed in the chloroplast and mitochondrion within its leaf cells, but separation appeared at the cytoplasm and the cell wall within the root cells. At ＞250 μmol/L Cu, both root and leaf organelles in E. splendens were damaged heavily by excessive Cu in vivo. Copper subcellular localization in the plant leaf after 8 days' exposure to 500 μmol/L Cu using gradient centrifugation techniques was found to be decreased in the order: chloroplast＞cell wall＞soluble fraction＞other organelles. The plant root cell wall was found to be the site of highest Cu localization. Increase of Cu exposure time from 8 d to 16d, increased slightly Cu concentration in cell wall fraction in roots and leaves, while that in the chloroplast fraction decreased in leaves of the plants grown in both 0.25 μmol/L and 500 μmol/L Cu. TEM confirmed that much more Cu localized in cell walls of E. splendens roots and leaves, but also more Cu localized in E. splendens' chloroplast when the plant is exposed to Cu levels＞250μmol/L, as compared to those in the plant grown in 0.25 μmol/L Cu. Copper treatment at levels＞250 μmol/L caused pronounced damage in the leaf chloroplast and root organelles. Copper localization in cell walls and chloroplasts could mainly account for the high detoxification of Cu in E. splendens.