OBJECTIVE:To establish a capillary zone electrophoresis with high frequency conductance method for the determination of ursolic acid in Radix et Rhizoma Clematidis.METHODS:The separation of sample was performed on an uncoated fused silica capillary(60 cm?75 ?m ID,effective length:56 cm) with 1.2 mmol?L-1 triethylamine-HCl(pH 10.60) and 0.24 mmol?L-1 ?-cyclodextrin as running buffer solution.The separation voltage was 12.5 kV and the gravity injection of sample was conducted for 10 s at a height of 20 cm.RESULTS:The calibration curve of ursolic acid was linear over the concentration range of 5.55～111.0 mg?mL-1(r=0.999 3) and the average recovery for ursolic acid was 96.0%(RSD=1.39%,n=6). CONCLUSION:The method was proved to be simple,accurate,rapid and reproducible,and suitable for the quantitative determination of ursolic acid in Radix et Rhizoma Clematidis.

Objective To summarize our experiences in the diagnosis and treatment of traumatic splenic rupture (TSR), in order to improve the diagnosis and treatment effect of TSR. Methods Retrospective (analysis) of the diagnosis and treatment of 184 patients with traumatic splenic rupture in recent 9 years was made. Results The preoperative correct diagnosis rate was 96.7% and was established on the history of (injury), clinical presentation, abdominal paracentesis, abdominal ultrasonography and CT. All the 34 of (patients) treated nonoperatively were cured. Of the 150 patients treated by operation, two died during operation and 148 patients were cured. Conclusions Combination of obtaining a detailed history of injury, physical examination, abdominal paracentesis, abdominal ultrasonography and CT can improve the accuracy rate of (preoperative) diagnosis.Under the ensurrance of the safety of the patients' life, preservation of the spleen should be performed if possible, especially for children. Both splenorrhaphy with or without ligation of splenic artery are simple, safe and effective methods to salvage the spleen.

OBJECTIVE:To prepare an intramyocardial bilayered porous biodegradable drug delivery stent and to evaluate its effects on myocardial channel after transmyocardial revascularization (TMR).METHODS:A biodegradable drug delivery stent was prepared by using poly (ε-caprolactone) (PCL),bovine serum albumin (BSA)and poly (D,L-lactide-co-glycolide) (PLGA).The levels of BSA in stent and released in vitro were determined by the Coomassie brilliant blue assay.The mechanical strength of stent was tested by universal material testing machines.Porcine models of chronic myocardial ischemia were created to evaluate the effects of this stent on myocardial channel after TMR,RESULTS:Each bilayered porous stent could carry 10 mg BSA and release about 80% of BSA after 30 days.The stent diminished 80% of initial scale under the stress of 1.2 MPa.It could keep myocardial channel patency after TMR.CONCLUSION:An intramyocardial bilayered porous biodegradable drug delivery stent was successfully prepared.It could sustain the pressure from the heart and maintain myocardial channel patency after TMR.

Objective To investigate the predictive value of 4 183 Da peptide of dermcidin protein in the early diagnosis and differential diagnosis of ischemic heart disease.Methods A prospective controlled study was conducted.Serum samples were drawn from 161 patients with acute coronary syndrome [ACS,including 46 patients with unstable angina (UA),23 with acute non-ST elevation myocardial infarction,and 92 with acute ST segment elevation myocardial infarction],111 subjects for routine physical examination,including 45 patients with hypertension history,42 with coronary heart disease,22 with diabetes,and 54 patients with non-ACS (including pulmonary embolism,aortic dissection aneurysm,arrhythmia,myocarditis,coronary myocardial bridge,pleurisy,pneumothorax,pneumomediastinum,rib fracture,reflux esophagitis,peptic ulcer,and pancreatitis) to serve as controls.4 183 Da peptide of dermcidin protein was assessed with matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology,and myeloperoxidase [MPO,determined by point-of-care testing (POCT) and enzyme linked i mmunosorbent assay (ELISA),respectively],high sensitive C-reactive protein (hs-CRP),heart type fatty acid binding protein (H-FABP),myoglobin (MYO),cardiac troponin Ⅰ (cTnⅠ),and MB isoenzyme of creatine kinase (CK-MB) were quantitated with biochemical analysis.The power of the biomarkers above for early diagnosis and differential diagnosis for ischemic heart disease were judged by comparison of their sensitivity and specificity.Results ① It was showed by one-way ANOVA that 4 183 Da peptide was higher in ACS group than that in control group (relative abundance:22.05 ± 16.97 vs.15.52 ± 14.09,P =0.001),but no difference was found between ACS group and non-ACS group (relative abundance:22.05 ± 16.97 vs.19.99 ± 17.63,P =0.416).② The specificity and sensitivity of the 4 183 Da polypeptide and MPO for predicting ACS and UA were compared with the receiver operating characteristic curve (ROC).It was showed that the 4 183 Da polypeptide had predictive values for ACS and UA,and the areas under the ROC curve (AUC) was 0.625 and 0.651 (both P ＜ 0.01),but MPO was not found to have predictive value (AUC was 0.440 and 0.336,respectively,both P ＞ 0.05).③ It was showed by the values of multi-markers in differential diagnosis of ACS and non-ACS disease that the specificity and sensitivity of 4 183 Da peptide in the differential diagnosis of acute myocardial infarction (AMI) and non-ACS disease were less than those of MYO,cTnⅠ,H-FABP,markers of myocardial damage,which AUCs were 0.569 vs.0.796,0.833,0.838,and equal to MPO (POCT/ELISA) and hs-CRP,AUC of which was 0.569 vs.0.505 (POCT)/0.477 (ELISA) and 0.545.But both the value of 4 183 Da peptide and MYO,cTnⅠ,H-FABP in the differential diagnosis of UA and non-ACS disease was not found,where AUC was 0.456,0.525,0.658,0.568.Conclusion 4 183 Da polypeptide,a fragment of dermcidin protein,may have association with the onset of ischemic heart disease,and may be helpful in the early diagnosis of ACS.

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Liver ; drug effects ; enzymology ; Metabolome ; Paeonia ; chemistry ; Rats ; Tandem Mass Spectrometry

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Objective To study the detection rate of contrast-enhanced ultrasound (CEUS) with different perfusion methods on rabbit VX2 small hepatocellular carcinoma less than 10.0 mm. Methods VX2 tumor cells were inoculated into the subcutaneous tissue of New Zealand rabbit′s thigh. Then the plant tumor were cut into small blocks under sterile conditions and transplanted into hepatic parenchyma in 30 New Zealand rabbits. The contrast media was injected through peripheral vein by single and double perfusion methods. The detection rate of two methods were compared. Results There were 41 hepatocellular carcinoma lesions in the 30 rabbits. There were 15 lesions with size between 3.0 mm and 5.0 mm, and 26 lesions between 5.0 mm and 10.0 mm in diameter. On CEUS, the VX2 tumor presented fast-in and fast-out pattern. In arterial phase, the lesion was enhanced rapidly. In portal venous phase, contrast began to wash out from the carcinoma. In delay phase, the enhancement of lesion was signiifcantly lower than the surrounding normal liver parenchyma. A total of 32 lesions were detected by single perfusion method, including 7 lesions ranging 3.0-5.0 mm and 25 lesions ranging 5.0-10.0 mm. A total of 39 lesions were detected by double perfusion method, including 13 lesions ranging 3.0-5.0 mm and 26 lesions ranging 5.0-10.0 mm. The detection rate of micro-hepatocellular carcinoma by single and double perfusion method was 78% and 95% respectively. The difference was statistically signiifcant (χ2=5.150, P=0.023). The detection rate of 3.0-5.0 mm lesions by single and double perfusion method was 47%and 87%, respectively. The difference was statistically signiifcant ( χ2=5.400, P=0.025). The detection rate of 5.0-10.0 mm lesions by single and double perfusion method was 96% and 100%, respectively. There was no statistically signiifcant difference (χ2=1.020, P=0.500). Conclusion The double perfusion method greatly promotes the detection of micro hepatocellular carcinoma, especially for the lesions less than 5.0 mm in diameter.

BACKGROUND:When peri-implantitis occurs, the levels of interleukin-17 in the peri-implant sulcular fluid and soft tissues are significantly increased, and there is a positive correlation with the depth of the peri-implant probing. However, because of the dual effects of bone absorption and protection, the function of interleukin-17 in peri-implantitis remains controversial, which should be further studied. OBJECTIVE:To retrospectively analyze the correlation between peri-implantitis and interleukin-17. METHODS: The first author searched Wanfang, CNKI and Medline databases from 1991 to 2014 by using key words of “dental implants, peri-implantitis, periodontitis, Th17 cels, interleukin-17, cytokines” in Chinese or English. The functions of bone destruction and protection of interleukin-17 in peri-implantitis were reviewed, and interleukin-17 expression and correlation with the peri-implantitis were detected. RESULTS AND CONCLUSION:According to the inclusion and exclusion criteria, 46 articles from 241 articles are selected. The results show that, noninvasive detection of the variation of interleukin-17 levels in peri-implant sulcular fluid or blood is conducive to the monitoring of early diagnosis and treatment of peri-implantitis. At present, most researches show the relationship between interleukin-17 and peri-implantitis, and there is a positive correlation between interleukin-17 and peri-implant probing depth. But other researches suggest that interleukin-17 acts as protector for bones, and there is no statistic significance in the level of interleukin-17 receptor gene in chronic peri-odontitis and healthy subjects. Therefore, the correlation between peri-implantitis and interleukin-17 remains to be further studied.

Anopheles dirus was fed with the mice of Kunming strain infected by plas-modium yoelii yoelii (By265 strain) for three or four days. Then the midgut and the salivary glands of the mosquitoes were examined. It was found that the infection rate of the malaria parasite of the midgut was 78.57% (44/56), but that of the salivary glands was only 4.88% (2/41) . After two passages of the malaria parasite through the mice and the mosquitoes, the infection rate of the salivery glands could be increased up to 13.96% (6/43).

Anopheles dims Peyton et Harrison, 1979 is an important malaria vector in Southeast Asia and China particularly in forested hilly zones. To establisha a laboratory mating colony for this mosquito is one of the subjects to be solved urgently in malaria research.This paper is to report our experience on breeding this mosquito. Ecological environment is created to simulate the natural conditions. The room-temperature of the insectary is 25?-28℃ and its relative humidity is around 80%.Mosquitoes are kept in cages with the si2e of 50 ? 50 ? 100cm. The larvae are fed with a mixture of de-fatted pig liver powder and yeast with a ratio of 1:3. The adults are fed with a mixture syrup containing 4% of orange juice, 10% of sucrose and other materials. The insectary is illuminated with a 3o w fluorescent light in the day time and a 15 w blue light in the night continuously to induce mating.The mating rate of the fourteenth generation reached 68% and the descendants bred can be used in research work now.