OBJECTIVE:To study the inhibitory effects of minocycline (MC) on TNF-α induced monocyte-endothelial adhe-sion and the relative mechanism. METHODS:Primary human umbilical vein endothelial cells (HUVECs) were isolated from hu-man umbilical veins with enzyme digestion. HUVECs were divided into blank control group,model group,MC low-dose,medi-um-dose and high-dose groups(1,10,100 μmol/L). After treated for 2 h,10 ng/ml TNF-α was employed to stimulate monocytes THP-1 adhesion with HUVECs except for blank control group,in order to induce monocyte-endothelial adhesion model. The num-ber of adherent cell was observed by fluorescence microscope,and fluorescence intensity was detected by microplate reader. Flow cytometry was adopted to detect the expression of intercellular adhesion molecule (ICAM)-1. The expressions of NF-κB p65 pro-tein in cell nucleus and cytoplasm were detected by Western blot. RESULTS:Compared with blank control group,the number and fluorescence intensity of adherent cell,the expression of ICAM-1 and the protein expression of NF-κB p65 in nucleus were all in-creased in model group,while the protein expression of NF-κB p65 in cytoplasm was weakened,with statistical significance(P<0.01). Compared with model group,the number and fluorescence intensity of adherent cell,the expression of ICAM-1 were all de-creased in MC low-dose,medium-dose and high-dose groups;the protein expression of NF-κB p65 in nucleus was weakened in MC medium-dose and high-dose groups,while the protein expression of NF-κB p65 in cytoplasm was heightened,with statistical significance (P<0.01 or P<0.05). CONCLUSIONS:MC can inhibit TNF-α induced monocyte-endothelial adhesion by a likely mechanism of reducing the expression of ICAM-1 in HUVECs and inhibiting the expression of NF-κB p65 protein.

The qualitative and quantitative analysis of active constituents in Fructus Psoraleae is presented by high-performance liquid chromatography (HPLC) coupled with different detections.Extracts of Fructus Psoraleae were examined by HPLC with ion trap mass spectrometry (IT-MS) and 18 major compounds of coumarins,benzofuran glycosides,flavonoids,and meroterpene were identified.The determination of four major constituents including bavachin,isobavachalcone,bavachinin,and bakuchiol was accomplished by HPLC with UV,MS,and electrochemical detection (ECD).These methods were evaluated for a number of validation characteristics (repeatability,LOD,calibration range,and recovery).ECD obtained a high sensitivity for analysis of the four components; MS provided a high selectivity and sensitivity for determination of bavachin,isobavachalcone,and bavachinin in negative-ion mode.After optimization of the methods,separation,identification.and quantification of the four components in Fructus Psoraleae were comprehensively tested by HPLC with UV,MS,and ECD.

Objective To investigate the degree of health knowledge and needs for health education of urban and rural inhabitants in Tianjin, and to study strategy of health education and health promotion in community. Methods Using a questionnaire survey, 900 inhabitants were investigated on their health knowledge, behavior, access to knowledge, and health education needs. Results The rate of health knowledge awareness was 65.63%, the rate of health promotion was 67.05%. Logistic regression results showed that the education background level (OR=1.323, 95%CI=1.113～1.573) and income per capita monthly(1.607, 1.317～1.962)were the influential factors for the rate of health knowledge awareness (P

Objective To evaluate lower extremity deep venous hemodynamic changes and lower extremity deep venous disease in abdominal obesity.Methods To compare venous flow parameters of the lower limbs as assessed by Duplex ultrasound scanning in obese and nonobese individuals according to body mass index(BMI).Venous hemodynamics were studied in a prospective cohort study in nonobese( BMI ＜ 25 kg/m2) and obese individuals( BMI ＞ 30 kg/m2 ).Diameter,peak,mean,and minimum velocities were assessed.Results The study examined 45 limbs in 24 nonobese individuals and 44 limbs in 22 obese individuals.The diameter of the femoral vein was significantly greater in obese vs nonobese limbs ( 10.4 ± 1.3) mm vs (7.1 ± 1.0) mm,P ＜ 0.01 ).Calculation of venous amplitude and shear stress showed significantly higher values in nonobese vs obese limbs [ ( 11 ± 5 ) vs ( 8 ± 3 ) cm/s ],( P ＜ 0.01 ) and [ (0.50 ± 0.17 ) vs ( 0.30 ± 0.09 ) ] dyn/cm2,( P ＜ 0.01 ).Spearman rank correlation revealed a significant inverse correlation between waist-to-hip ratios and waist circumference and velocities amplitude (peak ve locity-minim velocity),and shear stress.Conclusions Lower limb venous flow parameters differ significantly between obese and nonobese individuals.These findings suggest the mechanical role of abdominal adipose tissue potentially leading to elevated risk for both chronic venous insufficiency and venous thromboembolism.

Background Animal model of fungal keratitis is an available tool to the experimental study of the pathogenesis mechanism of fungal keratitis. Current modeling methods of fungal keratitis include corneal scratching, corneal stroma injection and corneal surface lens methods. But these methods still have their own shortages. Objective This experiment was to create a fungal keratitis animal model by modifying corneal surface lens method. Methods Modified animal models of fungal keratitis were created by modified corneal surface lens method in 12 general adult New Zealand white rabbits. The filter papers soaked 108 spores / ml or A106spores / ml of spergillus fumigatus suspension were attached on the de-epithelial cornea surface and fixed with contact lens and tarsorrhaphy for 2 days, and the filter paper with physiological saline was used as control group. The symptoms of anterior segment were examined under the slit lamp in 3 ,7 and 14 days after surgery and scored based on the criteria of Dong. Corneal scraping was stained with 10% potassium hydroxide and calcofluor white stain to observed mycelium under the fluorescence microscope. Corneal tissue sections were examined by hematoxylin-eosin staining and periodic acid Schiff staining under the light microscope. The use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results Fungal keratitis models were successfully established in 6 eyes and 4 eyes in 108 spores/ml group (6/6) and 106 spores/ml group respectively. The symptom was more severer and score was higher in the eyes of 108 spores/ml group than that in 106 spores/ml group. At 3 and 7 days after surgery,the symptom scores of fungal keratitis models were higher than those of control group from 3 through 7 days with the statistically significant difference (P<0. 01) and the symptom scores of 108 spores/ml group were significantly higher than those of 106 spores/ml group (P<0. 01). At 14 days after surgery, the symptom scores of 108 spores/ml group were still higher than those of control group (P<0. 05). Fungal hyphae was seen in the corneal scrapes in 108 spores/ml group and 106 spores/ml group respectively from 3 through 7 days after surgery. Inflammatory cell infiltration, stroma cells necrosis and fungal hyphae were presented in 108 spores/ml group, and the corneal neovascularization could be observed in 108 spores / ml group 14 days later. Fungal culture revealed the positive outcome in both 3 and 7 days after surgery in 108 spores/ml group,but in 106 spores/ml group,the positive result was only in the 3rd day. Conclusion Modified corneal surface lens method is more feasible and sample in the model of Aspergillus keratitis. This animal model of Aspergillus keratitis is practical for the further study of fungal keratitis.

Objective To determine the optimal cut-off value of critical-care pain observation tool (CPOT) in assessing degree of pain in patients undergone craniotomy, and to determine the sensitivity and specificity of CPOT with this cut-off value. Methods A prospective observational study was conducted in Beijing Tiantan Hospital. A total of 118 patients admitted to intensive care unit (ICU) after craniotomy was consecutively enrolled during August 2014 to August 2015. CPOT and visual analogue scale (VAS) were used to assess the pain before, during and 20 minutes after the removal of central venous catheters, and the difference was compared between two scores at three time points. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off values for evaluation of the sensitivity and specificity of CPOT. Patients' complaint of pain was considered the gold-standard. Results CPOT values (inter-quartile range) before, during and after the procedure were 0 (0-3), 0 (0-6) and 0 (0-2), respectively; while VAS values were 4 (1, 6), 3 (1, 6) and 4 (1, 6), respectively. CPOT value during the procedure was significantly higher than CPOT values before and after the procedure (both P < 0.01). When the optimal cut-off value of CPOT was 1, CPOT showed the highest Youden index before, during and after the procedure (1.183, 1.515, and 1.438, respectively), and showed high specificity (all 100%) and low sensitivity (18.3% and 43.8%, respectively) when assessing the pain before and after the removal of the catheter. The sensitivity and the specificity were high when assessing the pain during the procedure, the sensitivity was 69.4%, and the specificity was 82.1%. When the optimal cut-off value of VAS was 2 before and during the procedure, and was 4 after the procedure, VAS showed the highest Youden index, 1.568, 1.452, and 1.509, respectively. VAS demonstrated high sensitivity and specificity before, during and after the procedure (sensitivity was 97.2%, 95.2% and 75.0%, respectively; specificity was 59.6%, 50.0% and 75.9%, respectively). The area under ROC curve (AUC) of CPOT before, during and after the procedure were 0.592 [95% confidence interval (95%CI) = 0.490-0.693], 0.778 (95%CI= 0.693-0.863) and 0.719 (95%CI = 0.627-0.811), respectively; the AUC of VAS before, during and after the procedure were 0.846 (95%CI = 0.771-0.920), 0.767 (95%CI = 0.681-0.854) and 0.838 (95%CI = 0.767-0.909), respectively. The AUC of VAS before and after the procedure was significantly higher than the AUC of CPOT (P < 0.001 and P = 0.006), while there was no significant difference between the AUC of VAS and CPOT during the procedure (P = 0.826). Conclusion CPOT can be used to assess the pain during painful procedure, and it shows high accuracy, but with poor evaluation effect on pain in rest.

A strain of resistance to copper and cadmium with high concentration, named NTG-01, was isolated from soils of DaYe county mineral area in HuBei province. It can resist copper of 4.5mmol/L and cadmium of 2mmol/L , so we can say that it is a important strain used to study the resistance mechanism of copper and cadmium. A series of morphological and biochemical characteristics and sequences analysis of 16S rDNA reveal that it belongs to the bacteria and is gram negative, short rod, flagella around, the size of bacteria is about 0.8?m?2.0?m , V-P result shows positive, methyl-red result displays negative, and glucose can be utilized to produce acid and gas; In addition to, we find that it has the percent 99 homologous to Enterobacteraerogenes by 16S rDNA sequences BLAST analysis, plus the results of morphological and biochemical parameters, it belongs to Enterobacteraerogenes. We can conclude that NTG-01 has higher resistance to many different heavy metals by measuring MICs values of nine heavy metals at last.

As the components of proteoglycans, glycosaminoglycans (GAGs) are linear polysaccharides consisting of hexose and uronic acid units linked by β-1,3-glycosidic bond. GAGs mainly distribute in extracellular matrix and on cell surfaces. They guide many biological processes, such as proliferation of cells, transmission of signals and mediation of inflammation. Because of their large molecular weights, GAGs have limited biological functions in vitro. However, the appearance of chondroitinase ABC (ChSase ABC), which can lyse polysaccharides, solves the difficulties. Based on our work, we summarized the classification and the crystal structure of ChSase ABC, as well as other recent research progress on ChSase ABCs. The separation and purification methods of ChSase ABC and construction of engineering bacteria are illustrated. The stability and immobilization are also analyzed by taking account of the characterization of ChSase ABC. Finally, problems and future prospect of the ChSase ABC study are summarized.
Bacteria ; Chondroitin ABC Lyase ; chemistry ; Extracellular Matrix ; chemistry ; Glycosaminoglycans ; chemistry ; Proteoglycans ; chemistry

Objective To investigate whether the effect of pioglitazone on TNF-α-induced insulin resistance is associated with altering IRS-1-induced signaling. Methods 3T3-L1 adipocytes were treated with TNF-α for 24 hours with or without being pretreated with 10μM piglitazone for 6 hours or with pioglitazone alone.Insulin-stimulated glucose uptake of 3T3 adipocytes was measured by using 2-deoxy 3H glucose.The Western blot was used to measure IRS-1, PKB, PKC-λ protein and tyrosine phosphorylation on IRS-1, PKB and PKC-λ phosphorylation. Results Both TNF-α and pioglitazone increased glucose uptake of 3T3 adipocytes under basal status.On TNF-α treated cells, insulin-stimulated glucose uptake was decreased by about 50%, accompanied with the reductions of IRS-1 protein level, tyrosine-phosphorylation of IRS-1 and PKB phosphorylation.TNF-α treatment had no effect on PKC-λ phosphorylation. Pioglitazone pretreatment was able to antagonize TNF-α-induced insulin resistance in 3T3 adipocytes partly reverse IRS-1 protein, increase insulin-stimulated tyrosine phosphorylation of IRS-1,and increase phosphorylations of PKB and PKCλ. Conclusion TNF-α-induced insulin resistance in 3T3-L1 adipocytes is related to impaired tyrosine phosphorylation of IRS-1. Pioglitazone antagonizes the above TNF-α induced insulin resistance.

OBJECTIVE: To establish the method for rapid identification of the quality of Cortex moutan radicis and similar aromatic Chinese herbs by headspace gas chromatography (HSGC). METHODS: The constituents of headspace gas from Chinese herbs, such as Cortex moutan radicis,Radix cynanchi paniculati and root-bark of Paeonia lactiflora. were analyzed by HSGC. RESULTS: The characteristic finger-print headspace chromatograms of these three drugs were different from one another. Cortex moutan radicis stored for 4 years under common conditions contained only traces of paeonol. CONCLUSION: HSGC was simple, quick and accurate in identifying and controlling the quality of aromatic Chinese herbs.