OBJECTIVE A new piezoelectric aptamer biosensor is developed for determination of IgE. The energy converters are 10MHz AT-cut quartz crystals with gold-coated electrodes. The anti-IgE aptamers are immobilized onto the surfaces of crystals gold electrodes by biotin-avidin method. METHODS The standard substance and serum were detected to find the limit of detection and specificity of the biosensor. RESULTS The piezoelectric immunoglobulin aptamer biosensor could complete the detection without nonspecific response. Under the optimized conditions, the experimental results showed that the piezoelectric biosensor had good response to IgE whose frequency shifts were linearly dependent on IgE concentration in different range. The piezoelectric aptamer biosensor had been used to detect IgE in serum, the analytical results given by this method were in satisfactory agreement with those given by chemoluminescence method, its correlation coefficient was 0.9924. CONCLUSIONS Piezoelectric aptamer biosensor for the determination of IgE is of high sensitivity, high specificity, high analysis speed, unnecessary labeling, simple operation, real-time detection, etc. It is suitable for detection of IgE and should be used for clinical detection.

Objective To compare 2 methods to immobilize probes on the surface of gold membrane and select the better one for aptamer immobilization of aptamer-based piezoelectric quartz crystal sensor. Methods Anti-IgE aptamers were immobilized on the gold surface of piezoelectric quartz crystal sensor with thiol method or biotin-avidin method. The changes of frequency were compared in reactions with different concentrations of IgE. Results Biotin-avidin method obtained higher frequency changes in the reactions of IgE, lower limit of detection and wider liner range than thiol method. For biotin-avidin method, there was significant linear correlation between frequency changes and the concentration of IgE with the correlation coefficient of 0.994 5. Conclusion The thiol method which is proved effective for probe immobilization on gene sensor is unfit for aptamer immobilization. Biotin-avidin method has a better ability to immobilize probe on the surfaces of aptamer-based piezoelectric quartz crystal sensor.

OBJECTIVE To study the optimal reagent for regeneration of aptamer-coated piezoelectric quartz crystal chips and detect storage ability of aptamer-coated chips. METHODS Anti-IgE aptamers were immobilized on the gold surface of piezoelectric quartz crystal biosensor with biotin-avidin method. The reaction between anti-IgE aptamers and IgE was monitored in time on piezoelectric sensor. We tried to regenerate the sensor surface after binding IgE by rinsing the aptamer-coated chips with HCl, NaOH, EDTA, urea and formamide individually. Aptamer-coated chips were stored in PBS with 0.1% sodium azide and the stored chips were used to detect IgE in 35 days. RESULTS Of the five reagents, EDTA was the best one for regeneration of aptamer-coated chips and the sensor could retain 91.5% of the original detecting signals after five regeneration cycles. Moreover, the aptamer-coated chips could be stored in the binding buffer for 21 days without obvious loss of activity. CONCLUSIONS Compared with antibody-based piezoelectric sensor, aptamer-based sensor has lower cost, more regeneration cycles and longer time for the storage of chips. This series of experiments shows the superiority of aptamer detection.

OBJECTIVE To establish an aptamer-based piezoelectric sensor assay to detect human immunoglobulin E (IgE) and acquire the parameters about this kind of biosensor. METHODS Anti-IgE aptamers were immobilized on the gold surface of piezoelectric quartz crystal sensor with biotin-avidin method. The reaction between anti-IgE aptamers and IgE was monitored in time on piezoelectric sensor. Different concentrations of IgE were detected and the calibration curve of IgE was drawn. Non-specific response signals of BSA, IgG and IgE were monitored to show the detecting specificity of aptamer-based sensor. RESULTS For the aptamer-based sensor, the lowest limit of detection was 0.055mg/L and the linear range was 0.1-2.5mg/L. Non-specific signals of this sensor were less than 5% of specific signals of IgE at the same concentration. CONCLUSIONS The aptamer-based piezoelectric sensor can detect 5.5 ng IgE while the non-specific signals are very low. So this aptamer-based sensor is hopeful to be applied to clinical laboratory diagnosis.

This study was aimed to verify the effects of staged sequential therapy on expressions of matrix metalloproteinase-9 (MMP-9) and its inhibitor TIMP-1 within lung tissues in asthmatic rats with the airway remodeling, by applying a series of tests such as the immunohistochemistry, western blot and real-time fluorescent quantitative PCR. SD rats were randomly divided into 7 groups, which were the asthmatic group (Group X), the normal group (Group Z), the No. 1 sequential therapy group (Group A1), the No. 2 sequential therapy group (Group A2), the No. 3 sequential therapy group (Group A3), the montelukast group (Group M), and the budesonide group (Group B). The asthmatic model was established in each group except Group Z, by sensitization with both ovalbumin (OVA) and aluminium hydroxide via injection at the 1st, 8th and 15th day in a 22-day duration, followed by OVA aerosol inhalation every other day for 8 weeks for asthma activation. At the 8th day after the asthmatic model was established, Group A1 was orally given Ma-Xing Er-Chen Tang (MXECT) during acute phase while given normal saline (NS) during catabasis and stable phase; Group A2 was given MXECT during acute phase, and given modified Jin-Shui Liu-Jun Jian (JSLJJ) during catabasis as well as given NS during stable phase; Group A3 was given MXECT during acute phase, and given modified JSLJJ during catabasis as well as given Liu-Wei Di-Huang (LWDH) Powders during stable phase;Group M was given salbutamol via aerosol inhalation after stimulation, while orally given montelukast during catabasis and stable phase; Group B was given salbutamol via aerosol inhalation after stimulation, while given inhaled budesonide during catabasis and stable phase; Group X was given NS. After the 7-week intervention, the immunohistochemistry, western blot and real-time quantitative PCR were applied to analyze the location and quantitative expression of MMP-9 and TIMP-1, so as to find out the biological mechanism on expressions of MMP-9 and TIMP-1 in lung tissues of asthmatic rats from molecular levels to gene levels. The results of immunohistochemistry, western blot and real-time fluorescent quantitative PCR showed as follows. Compared with Group Z, the contents of MMP-9 and TIMP-1 increased significantly within lung tissues in Group X. Compared with Group X, the contents of MMP-9 and TIMP-1 decreased within lung tissues of asthmatic rats in each treatment group. It was concluded that the expression of MMP-9 and TIMP-1 elevated during asthma. TCM staged sequential therapy can regulate the ratio between MMP-9 and its inhibitor so as to block the airway remodeling, by decreasing the expression of MMP-9 and its inhibitor within lung tissues in asthmatic rats. This is one of the important action mechanisms.

Objective To explore the clinical efficacy of Shenkang Injection and its influence on C-reactive protein (CRP)and interleukin-6 (IL-6)levels in patients with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus combined with DN admitted in our hospital from Jan.2012 to Jan.2014 were randomly divided into observation group and con-trol group,60 cases for each.Control group was treated with high-quality protein diets and insulin to control blood glucose and pressure,on which basis observation group was added with intravenous injection of Shenkang Injection.Clinical efficacy,fasting blood glucose (FBG),Triglyceride (TG), total cholesterol (TC),serum creatinine (SCr),24 h urinary protein (24 hUpro),CRP and IL-6 level changes before and after treatment in both groups were observed.Results Clinical efficacy was 90.00% in observation group,evidently higher than 75.00% in control group (P <0.05). Aboveindexeswere all improvedobviously after treatment than treatment before (P < 0 .0 5 ,P <0.01)and were markedly lower in observation group than in control group (P <0.01).Conclusion Shenkang Injection can effectively reduce IL-6 and CRP levels and decrease blood glucose and pressure,prolong disease progression and improve prognosis in DN patients.

Objective To explore the clinical efficacy of Shenkang Injection and its influence on C-reactive protein (CRP)and interleukin-6 (IL-6)levels in patients with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus combined with DN admitted in our hospital from Jan.2012 to Jan.2014 were randomly divided into observation group and con-trol group,60 cases for each.Control group was treated with high-quality protein diets and insulin to control blood glucose and pressure,on which basis observation group was added with intravenous injection of Shenkang Injection.Clinical efficacy,fasting blood glucose (FBG),Triglyceride (TG), total cholesterol (TC),serum creatinine (SCr),24 h urinary protein (24 hUpro),CRP and IL-6 level changes before and after treatment in both groups were observed.Results Clinical efficacy was 90.00% in observation group,evidently higher than 75.00% in control group (P <0.05). Aboveindexeswere all improvedobviously after treatment than treatment before (P < 0 .0 5 ,P <0.01)and were markedly lower in observation group than in control group (P <0.01).Conclusion Shenkang Injection can effectively reduce IL-6 and CRP levels and decrease blood glucose and pressure,prolong disease progression and improve prognosis in DN patients.

Objective : To evaluate the effect of platelet-rich fibrin (PRF) on orthodontic tooth movement (OTM) and to provide a basis for clinical application. Methods: Literature searches were conducted in 7 electronic databases supplemented with a hand search. Randomized controlled trials focusing on OTM with PRF were included. The risk of bias was assessed by the Cochrane tool. Finally, due to the heterogeneity of patient clinical characteristics and research methods, the results in every study were qualitatively described. Results: Six studies were included. Five studies were split-mouth designs, and 1 was a two-arm parallel design. Two studies used leukocyte- and platelet-rich fibrin, while the other 4 used injectable PRF. The risk of bias of 3 studies was graded as “Some concerns”, and 3 were graded as “Low risk”. The trials lasted from 4 weeks to 5 months. Four studies supported that PRF could accelerate OTM, 1 study demonstrated that PRF had no effect on OTM, and 1 study reported that PRF decreased OTM. There is moderate-quality evidence that PRF accelerates OTM in the first 3 months after application, while low-quality evidence supports that PRF loses its tooth-acceleration effect after 4 months. Conclusion Limited clinical evidence suggests that PRF could accelerate OTM in the early stages, but its long-term effect needs clarification.

Objective: To investigate the role of endoplasmic reticulum stress (ERS) in rats with acute respiratory distress syndrome (ARDS) related right ventricular dysfunction and the protective effect of sodium 4-phenylbutyrate (4-PBA) on right ventricle. Methods: Sixty male Spragne-Dawley (SD) rats were randomly divided into control group (CON group), lipopolysaccharide (LPS) model group, 4-PBA prevention group and 4-PBA treatment group, with 15 rats in each group. ARDS rat model was established by intratracheal instillation of LPS 10 mg/kg after tracheotomy; CON group was given the same amount of saline. 4-PBA prevention group and 4-PBA treatment group were given 4-PBA 500 mg/kg intragastric administration 2 hours before and after LPS respectively. Echocardiography was performed 12 hours after treatment to evaluate the right ventricular function. Then, the rats were sacrificed by bloodletting, and the serum and right ventricular tissue were harvested. The histopathological changes of myocardial were observed by hematoxylin-eosin (HE) staining, the levels of tumor necrosis factor-α(TNF-α), interleukins (IL-1β and IL-6) in serum and myocardial were detected by enzyme linked immunosorbent assay (ELISA), and Western Blot was used to detect the expression of the marker proteins of ERS in myocardial, including glucose regulatory protein 78 (GRP78), C/EBP cyclic adenosine phosphate reaction primitive binding transcription factor homologous protein (CHOP), caspase-12 and caspase-3. Results: Compared with the CON group, the echocardiography showed pulmonary artery maximum pressure gradient (PAmaxPG), pulmonary artery acceleration time (PAAT), tricuspid annular plane systolic excursion (TAPSE) in LPS model group were significantly decreased, and right ventricular end-diastolic excursion (RVDd) was significantly increased, and the levels of TNF-α, IL-1β and IL-6 in serum and myocardial, as well as the expressions of GRP78, CHOP, caspase-12 and caspase-3 in myocardial were significantly increased. Compared with LPS model group, TAPSE of 4-PBA preventive and treatment groups were significantly increased (mm: 3.08±0.65, 2.96±0.61 vs. 2.48±0.45), RVDd were significantly decreased (mm: 3.67±0.58, 3.60±0.61 vs. 4.18±0.71), the levels of TNF-α, IL-1β and IL-6 in serum and myocardial were significantly decreased [TNF-α (ng/L): 187.98±18.98, 176.08±17.98 vs. 332.00±19.90 in serum, 135.06±19.00, 132.78±17.00 vs. 155.00±20.00 in myocardial; IL-1β(ng/L): 12.07±2.98, 11.05±2.41 vs. 24.06±4.01 in serum, 19.89±2.80, 21.06±2.80 vs. 26.00±2.60 in myocardial; IL-6 (ng/L): 42.98±7.90, 34.05±6.09 vs. 89.80±10.07 in serum, 129.45±25.00, 127.08±26.06 vs. 145.77±23.00 in myocardial]; the expressions of GRP78, CHOP, caspase-12 and caspase-3 in myocardial were significantly decreased (GRP78/GAPDH: 0.090±0.070, 0.103±0.060 vs. 0.167±0.090, CHOP/GAPDH: 0.109±0.090, 0.090±0.080 vs. 0.186±0.090, caspase-12/GAPDH: 0.769±0.230, 0.799±0.210 vs. 1.040±0.350, caspase-3/GAPDH: 0.391±0.060, 0.401±0.054 vs. 0.603±0.340), with statistically significant differences (all P < 0.05). There were no significant differences in each indexes between 4-PBA prevention group and 4-PBA treatment group (all P > 0.05). Conclusions ERS is involved in ARDS-related right ventricular dysfunction. 4-PBA can protect the right ventricular function of ARDS rats by inhibiting ERS and alleviating inflammation, and the preventive and therapeutic effects of 4-PBA are similar.