OBJECTIVE:To establish a method for the content determination of menthanol in Qinagli pipa syrup. METHODS:Headspace GC was performed on the column of HP-INNOWax capillary column,column temperature was 80 ℃,temperature inlet sample was 150 ℃,the detector was hydrogen flame ionization detector with temperature of 250 ℃,the flow rate of nitrogen was 0.8 ml/min with constant flow and no split,the volume injection was 1 ml,the equilibrium temperature of headspace bottle was 80 ℃,and the equilibrium time was 20 min. RESULTS:The linear range of menthanol was 2.486-24.86 μg/ml(r=0.9999),RS-Ds of precision,stability and reproducibility tests were lower than 2%,recovery was 95.4%-99.8%(RSD=1.5%,n=9). CON-CLUSIONS:The method is simple and accurate,and can be used for the content determination of menthanol in Qinagli pipa syrup.

Objective To study the method of headspace gas chromatography for the determination of β-eugenol in Magnolia officinalis. Methods Headspace gas chromatography was used. The column was HP-INNOWax capillary column, the detector was hydrogen flame ionization detector, inlet temperature: 200 ℃; column temperature: 135 ℃ for 3min, at 10 ℃/min heating rate rose to 150 ℃, At a rate of 15 ℃/min up to 250 ℃for 5 min; constant flow rate: 0.8 mL/min; detector temperature: 250 ℃; headspace injection, gas injection volume: 1 mL, headspace Temperature 180 ℃, the balance time 30min; with the external standard method according to the peak area calculation results. Results The linear range of the concentration of menthol was 2.49～24.86 μg/mL (r=0.9994). The RSD of the precision, stability and repeatability test were <5％, the average recovery was 97.0％, RSD was 0.84％ (n=9). Conclusion The method is simple and accurate for the determination of volatile oil.

Objective To evaluate the efficacy of adjustable oral appliance (OA) on ventilation of patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods Databases were searched from Junuary 1990 to July 2014 for randomized controlled trials on the use of adjustable OA for the promotion of ventilation in OSAHS.Meta analysis was performed with RevMan5.2 software,and descriptive analysis of the data that could not be converted and merged was performed.Results Totally 6 RCTs involved,including 1 109 patients,which involved 4 random parallel controlled trials and 2 random crossover controlled trials.Meta-analysis showed that adjustable OA were associated with a decreased apnea-hypopnea index(AHI) [fixed effect model,WMD=-2.38,95％CI (-3.59--1.17),P＜0.01] and an improvement in minimum arterial oxygen saturation (SaO2)[randomized effect model,WMD=3.09,95％CI(1.16-5.02),P＜0.01] of OSAHS patients.The descriptive analysis showed that the difference in the sleep score was not statistically significant because the data could not be converted.Conclusions Compared to fixed oral appliance,adjustable oral appliance produced greater effect in reduction of AHI and the improvement in minimum SaO2,but the sleep score is not improved significantly,the large sample and multi-center randomized controlled test needs to be designed for the verification of comprehensive effect of adjustable oral appliance.

Objective To investigate the predictive value of plasma cystatin C (CysC) in patients with acute coronary syndrome (ACS) after pereutaneous coronary intervention (PCI).Methods A total of 660 patients with ACS admitted to cardiovascular department were enrolled in this study from January 2009 to June 2010.The enrollment criteria were:(1) the stenosis degree was above 75％ in at least one coronary artery checked by coronary angiography and successful PCI; (2) normal renal function or mild dysfunction with glomerular filtration rate (GFR) ＞ 60 ml/ ( min · 1.73 m2 ).Exclusion criteria were severe liver and renal insufficiency,malignancies and valvular heart diseases.The plasma CysC levels were examined by the latex enhanced immune turbidity method within 24 hours after admission.The relevant clinical data were recorded.The patients were followed up by out-patient interview or telephone from March to June 2011 and adverse cardiovascular events were recorded.The patients were divided into four groups according to CysC level:Q1 (CysC＜1.02 mg/L),Q2 (1.02 mg/L≤＜CysC ＜1.17 mg/ L),Q3 (1.17 mg/L ≤ CysC ＜1.35 mg/L) and Q4 (CysC ≥ 1.35 mg/L).Univariate and multivariate Cox hazards regressions were established to analyze the factors related to prognosis.The proportion differences between four groups were tested by x2.The survival ratio was estimated using the Kaplan-Meier method.Statistical significance was established at a P value of less than 0.05.Results ① A total of 606 ( 91.7％ ) patients successfully accepted follow-up.Mean follow-up time was ( 14.3 + 1.7 ) months.Of them,95 patients were subjected to adverse cardiovascular events ( 15.7％ ).②The incidences of adverse cardiovascular events in Q2,Q3,Q4 were significantly higher than those in Q1 ( P ＜ 0.001 ).The rates of mortality,nonfatal myocardial infarction and target lesion revascularization in Q4 were higher than those in Q1 ( P ＜ 0.05 ).The incidences of heart failure in Q3 and Q4 were higher than that in Q1 ( P ＜ 0.05 ).③Univariate analysis demonstrated that CysC,creatinine,LVEF,age,history of PCI and NYHA grade ≥3 were the risk factors of poor prognosis (P ＜ 0.05 ).④ Multivarite cox hazards regression revealed that the elevation of CysC level remained an independent predictor of adverse cardiovascular events.The relative risk of Q3 and Q4 were 3.930 (95％ CI 1.306-11.829,P =0.015 ) and 6.380 (95％ CI 2.171-18.751,P =0.001 ) compared with Q1.⑤　The cumulative rates of survival without adverse cardiovascular events in Q2,Q3 and Q4 decreased compared with Q1 (P ＜ 0.001 ).Conclusions High plasma CysC concentration is an independent predictor of adverse cardiovascular events in patients with ACS after PCI.

BACKGROUND:Bone marrow mesenchymal stem cels are crucial for bone and cartilage development and regeneration at a celular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cels is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cels and disease progression in osteoarthritis patients.METHODS: Thirty patients with osteoarthritis were enroled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cels. Proliferation ability of cels at passage 3 was detected using cel counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION:Afterin vitro culture, bone marrow mesenchymal stem cels grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cels were in cel division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cels from the three groups were al at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cels were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cels from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients.

OBJECTIVE: To explore the effects of three dentine desensitizers on the surface morphology of freshly exposed dentin and to evaluate their occlusion effects on dentinal tubules using scanning electron microscope (SEM). METHODS: A total of 16 isolated human premolar samples, which were prepared to expose dentine, were randomly divided into a control group (n=4), a Hybrid Coat group (n=4), a Prime & Bond NT group (n=4), and a anti-sensitivity toothpaste group (n=4). After treatment with dentine desensitizers, one half of the samples in each group were vertically cleaved. Finally, the surfaces and cross sections of the samples were observed by SEM. RESULTS: The exposed tubule was almost occluded in the Hybrid Coat group and the Prime & Bond NT group, while only the majority of tubules could be sealed in the anti-sensitivity toothpaste group. The cross-section images showed that sediments were visible in all groups except the control group. CONCLUSION Hybrid Coat and Prime & Bond NT are able to effectively seal tubules, while the effect of anti-sensitivity toothpaste is slightly poor.
Dentin ; ultrastructure ; Dentin Desensitizing Agents ; pharmacology ; Dentin Permeability ; drug effects ; Humans ; Microscopy, Electron, Scanning ; Polymethacrylic Acids ; pharmacology ; Toothpastes ; pharmacology

Objective To evaluate the transfusion efficacy and safety of patients with non‐specificity irregular antibodies .Meth‐ods A total of 19 cases of patients with non‐specificity irregular antibodies were analyzed ,then the transfusion efficacy and the in‐cidences of adverse transfusion reactions were investigated .Blood donor samples with the same ABO blood type of patients were randomly collected ,then blood cross‐matching was carried out .And the transfusion efficacy and safety of patients with non‐specific‐ity irregular antibody were evaluated when compatible blood were found by using both micro‐column gel technique and saline meth‐od .Results The non‐specificity irregular antibodies of the 19 cases of patients all were IgG irregular antibodies .The erythrocyte transfusion was carried out in the 19 cases of patients ,the total efficiency rate ,partial efficiency rate and inefficiency rate was 73 .7% ,26 .3% and 0 .0% ,respectively .No delayed hemolytic transfusion reactions were observed in any patient .Conclusion For patients with non‐specificity irregular antibodies ,the blood transfusion could be efficacy and safety when compatible blood samples are confirmed by using both micro‐column gel technique and saline method .

Objective To investigate the predictive value of metabolic syndrome in patients with acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI).Methods A total of 660 patients with ACS admited to cardiovascular department,first affiliated hospital of zhengzhou university were enrolled in this study from January 2009 to June 2010.The enrollment criteria were:the stenosis degree were above 75％ in at least one coronary artery by coronary angiography and successful PCI procedure.Exculsion criteria were:liver and renal insufficiency,malignancies and valvular heart diseases.The relevant clinical data and labtory examination were recorded after admission. The patients were followed up by outpatients interview or telephone from March to June 2011 and adverse cardiovascular events were recorded.The patients were divided into MS and non-MS groups,and basic clinical data were compared between two groups.The proportion difference between two groups were tested by chi square. Multivariate logistic regression was established to analyze the factors related to progonosis.The survival ratio was estimated using the Kaplan-Meier method.Statistical significance was established at a P value of less than 0.05.Results ①A total of 606 (91.7％) patients successfully accepted follow-up.Mean follow-up time were ( 14.3 ±1.7 ) months.95 patients experienced adverse cardiovascular events ( 15.7％ ).②There were 393 patients (64.96％ ) satisfied the definition of metabolic syndrome.The patients in MS group were with higher BMI,SBP,DBP,blood glucose and disordered lipid (all P ＜ 0.05 ),with less fale patients (P =0.016),less current somking (P =0.008 ) and with higher platelet (P =0.037 ). The incidence of adverse cardiovascular events in two groups were 17.81％ and 11.79％ ( P ＞ 0.05 ). ③ Multivarite logistic regression revealed that the predictors of adverse cardiovascular events were age [ OR =2.628,95％ confidence interval (CI) 1.395 ～ 4.954,P =0.003 ],New York Heart Association (NYHA) ≥ 3 grade ( OR =2.310,95％ CI 1.095 ～4.870,P =0.028) and left ventricular ejection fraction (LVEF) ( OR =4.328,95％ CI 1.955 ～9.580,P ＜ 0.001 ).However,MS was not related with prognosis ( OR =1.170,95％ CI 0.583 ～ 2.345,P =0.659 ).④The cumulative survival rates of no adverse cardiovascular events in the two groups were no significant difference ( P ＞ 0.05 ).Conclusions MS is a risk factor with coronary heart disease.Howerer,it has no relationship with adverse cardiovascular events in patients with ACS after PCI.

Objective To construct a recombinant immunotoxin expression vector composed of a single-chain Fv fragment of Sehistosorna japomicum and PE38KDEL gene,and identify the binding activity of the purified product with SEA antigen.Methods The V_H and V_L genes were amplified by PCR from the parent monoclonal antibody NP11-4.Then the amplified scFv and PE38KDEL genes were inserted into the expression vector pBAD/gIII A.The fusion protein expressed in E.coli Top10F' induced by L-arabinose.After purification,the activity of the immunotoxin was evaluated by Westem-blot and ELISA.Results The new recombinant immunotoxin expression vector pBAD/gIII A-scfv-PE38KDEL was constructed successfully.The main product was in inclusion bodies.ELISA assay showed that the refolding recombinant immunotoxin remained binding activity with SEA antigen.Conclusion A new recombinant expression plasmid pBAD/gIII A-scfv-PE38KDEL has been constructed and expressed successfully,which is useful in further study of the treatment of schistosomiasis japonica.

Objective:To investigate the related mechanism of miRNA-34a (miR-34a) reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 (HMGB1).Methods:The MG-63 cisplatin-resistant cell line (MG-63/DDP) was constructed by using dose escalation and intermittent action, and then the successfully constructed MG-63/DDP cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate. The MG-63/DDP cells were transfected respectively and then randomly divided into two groups: the miR-34a overexpression vector group and the miR-34a empty expression vector (miR-34a-NC) group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-34a. Transfected cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis. The dual luciferase reporter gene experiment was used to verify whether miR-34a regulated the expression of HMGB1, and Western blotting method was used to detect the HMGB1 protein expression level of the transfected cells. MG-63/DDP cells were transfected respectively and then randomly divided into two groups: HMGB1 gene silencing vector (si-HMGB1) group and its negative control vector (si-NC) group. Western blotting method was used to detect HMGB1 protein expression level and CCK-8 method was used to detect cell survival rate.Results:The MG-63/DDP cell line was successfully constructed. The survival rate of MG-63/DDP cells was higher than that of MG-63 cells when cells were treated with different concentrations of cisplatin (all P < 0.01), and half inhibitory concentration ( IC50) value of MG-63/DDP cells and MG-63 cells on cisplatin was 25.80 μg/ml and 0.47 μg/ml, respectively. qRT-PCR results showed that the relative expression level of miR-34a in MG-63/DDP cells was lower than that in MG-63 cells (0.46±0.04 vs. 1.02±0.05, t = 15.14, P < 0.01); compared with miR-34a-NC group, the relative expression of miR-34a in MG-63/DDP cells was increased in miR-34a overexpression vector group (1.67±0.09 vs. 1.00±0.02, t = -12.58, P < 0.01). Cell survival rate of miR-34a overexpression vector group and miR-34a-NC group was decreased with the increase in the concentration of cisplatin; cell survival rate of miR-34a overexpression vector group was lower than that of miR-34a-NC group when cells were treated with different concentrations of 5- 30 μg/ml cisplatin (all P < 0.01). The apoptotic rate of MG-63/DDP cells in miR-34a-NC group and miR-34a overexpression vector group was (25.1±1.7)% and (42.3±2.3)%, respectively when cells were treated with 20 μg/ml cisplatin; and in miR-34a overexpression vector group, MG-63/DDP cells had a higher rate of apoptosis ( P < 0.01). The dual luciferase reporter gene experiment results showed that compared with miR-34a-NC group, miR-34a overexpression vector group could inhibit the luciferase activity of PGL3- wild-type-HMGB1, and the difference was statistically significant ( t = 6.37, P < 0.01), while miR-34a overexpression vector group had no significant inhibitory effect on the luciferase activity of PGL3- mutant-HMGB1 ( t = 0.35, P = 0.74). The relative expression level of HMGB1 protein in miR-34a overexpression vector group was lower than that in miR-34a-NC group (0.43±0.02 vs. 1.00±0.14, t = 6.98, P < 0.01). Compared with si-NC group, the relative expression level and IC50 value of HMGB1 protein in si-HMGB1 group were reduced (all P < 0.05). Conclusion:Overexpression of miR-34a can enhance the chemosensitivity of osteosarcoma cells MG-63/DDP to cisplatin, and its mechanism may be related to the inhibition of HMGB1 expression.