Objective: To develop a capillary electrophoresis system for purity test and enantioseparation of L-pramipexole. Methods: Using β-CD, HP-β-CD, SBE-β-CD and DM-β-CD as chiral selection agents, we investigated the influence of concentration, pH of chiral selection agent and column length and inner diameter on chiral separation. The enantiomers were baseline separated under the following conditions-uncoated fused silica capillary (50 μm X 47 cm) with an effective length of 40 cm, back-ground electrolyte solution, 40 mmol/l trisodium citrate buffer solution, 0.5% SBE-β-CD, pH 4.0 adjusted by phosphoric acid, 5% methanol, 25 mmol·L-1 SDS, 40 mmol·L-1 sodium tetraborate solution as running buffer, and 16 kV separation voltage. The detection wavelength was set at 254 nm and the column temperature was 30*C. Results: The baseline separation of L-pramipexole was obtained under our condition. The optical isomerism had satisfactory resolution (Rs>2.5) and the content of R-pramipexole was less than 0.5%. Conclusion: The proposed method is simple, reliable and can be used for routine purity test and chiral separation.

Objective: To investigate the expression of nuclear factor-κB (NF-κB), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2(TIMP-2) in the kidneys of rats with acute renal trauma, and to discuss the influence of ulinastatin on their expression and its protective mechanism on the kidney. Methods: The animal model was established by striking the rachi-costaz zone with falling object from the height of 45 cm. Sixty-six rats were randomly divided into three groups:normal control group (C,n = 6),trauma group (TRA,n = 30),and ulinastatin+trauma (UTI,n = 30); the last 2 groups were further divided into 1 h, 6 h, 12 h, 18 h and 24 h subgroups, with 6 animals at each time point. Tissue microarray and immunohistochemistry were used to examine the expression of NF-κB and MMP2/TIMP-2 in different groups. Results: MMP-2 and NF-κB began to express 1 h after trauma in TRA group and their expression was significantly stronger than that in the control group (P < 0.05, P < 0.01); their expression reached the peaks at 12 h and 6 h after trauma and then gradually decreased. The expression of MMP-2 and NF-κB in UTI group reached their peaks 18 h and 12 h after trauma,respectively,and was significantly higher than that in the control group(P<0.01),but was lower than that in the TRA group at corresponding time points(P<0.01,P<0.05). The expression of TIMP-2 was significantly stronger than that in the control group and TRA group at 6 h, 12 h and 18 h after trauma(P<0.01,P<0.05). Conclusion: The expression of NF-κB,MMP-2 is increased in acute traumatic tissue of the kidney; the increase of TIMP-2 is not evident. Ulinastatin can protect the kidney by inhibiting the expression of MMP-2 and NF-κB and maintaining the balance of MMP-2/TIMP-2.

OBJECTIVETo evaluate the clinical effect of arthroscopic excision of the os subfibulare in anterior-lateral ankle pain.

METHODSFrom December 2005 to Augest 2014, 16 patients suffering from pain associated with an os subfibulare in the anterior-lateral side of their ankles were reviewed. Among the patients,11 patients were male and 5 were female, with a mean age of (33.5 ± 15.6) years old. The mean maximum diameter of os subfibulare was (0.70 ± 0.26) cm. All the patients underwent excision of the osseous fragments, and had anatomic reconstruction of the anterior talofibular ligament if the anterior-lateral ankle was instable. The average follow-up period was (18.0 ± 4.5) months. To analyze the surgical outcome, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot pain and function scales,visual analogue scale (VAS) and Tegner activity scale were assessed preoperatively and postoperatively.

RESULTSAOFAS scales were preoperative 60.15 ± 14.52 and postoperative 92.35 ± 5.73. There was a significant difference between them (t = -8.251, P = 0.000). The mean VAS score were preoperative 7.35 ± 0.46 and postoperative 2.45 ± 0.98. Statistical significance was also notable (t = 18.105, P = 0.000). Tegner score was significantly increased from preoperative 2.87 ± 1.12 to postoperative 5.78 ± 1.06 (t= -7.548, P = 0.000).

CONCLUSIONIrrespective of the size of os subfibulare, in patients with pain or instability associated with the os subfibulare, arthroscopic excision combined with reconstruction of ther anterior talofibular ligament or not was effective in restoring ankle function and eliminating pain.

Adult ; Ankle Injuries ; surgery ; Ankle Joint ; surgery ; Arthroscopy ; methods ; Female ; Fibula ; surgery ; Humans ; Lateral Ligament, Ankle ; surgery ; Male ; Middle Aged

To study the effect of Fuzheng Huayu recipe (FZHY) on five types of isozymes of cytochrome P450 (CYP450) of normal and liver fibrosis rats by using the cocktail probe method. Dimethylnitrosamine ( DMN) was injected to induce the liver fibrosis model. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the plasma concentrations of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK solutions 2. After the oral administration with FZHY, normal rats given phenacetin, omeprazole, tolbutamide and dextromethorphan showed increase in AUC(0-t) and decrease in CL to varying degrees, indicating that FZHY obviously inhibited the activities of CYP1A2, CYP2C9, CYP2C19 and CYP2D6 in normal rats, but with no obvious effect on the activity of CYP3A4. After the oral administration with FZHY, liver fibrosis rats treated with CYP2C9 showed the significant increase in AUC(0-t) and significant decrease in Vd, hut with no obvious changes in the pharmacokinetic parameters of other four types of prove substances, suggesting that FZHY could significantly inhibit the activity of CYP2C9 in rats but had no effect on the activities of CYP1A2, CYP2C19, CYP2D6 and CYP3A4. The changes in the activity of CYP450 isozymes in liver fibrosis rats may be the reason for FZHY's different effects on CYP450 isozymes in normal and liver fibrosis rats.
Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Humans ; Isoenzymes ; genetics ; metabolism ; Liver Cirrhosis ; drug therapy ; enzymology ; genetics ; Male ; Mass Spectrometry ; Rats ; Rats, Wistar

Objective:To investigate the expression and prognostic significance of CD19 in patients with Acute myelogenous leukemia with AML1-ETO positive. Methods: Clinical data of 66 patients AML with AML1-ETO positive who were newly diagnosed from Jan 2010 to Dec 2015 were collected. To retrospectively analyze the relationship between clinical characteristics and expression of CD19,so dose the prognosis. Results:The positive rate of CD19 expressing in AML with AML1-ETO positive was 50. 0%. There were no statistically significant differences in terms of age,gender,hemoglobin,platelet,percentage of bone marrow blasts,accompanied with chromosome ,gene mutations between patients with and without CD19 expression(P>0. 05). The white blood cell count(WBC) of the CD19 negative group was higher than CD19 positive group,while showed significant difference(P=0. 027). Although the relapse-free survival (RFS) of patients with CD19 expression was higher than those without,no significant difference was calculated (P=0. 105). Patients with CD19 expression had superior overall survival ( OS ) compared to those without CD19 expression ( P = 0. 030 ) . Multivariable analysis for OS identified CD19 positivity as an independent predictor associated with better prognosis. Conclusion: The expression of CD19 in AML with AML1-ETO positive may be an indicator associated with better prognosis.

The teaching status of Molecular Pharmacognosy in 28 institutions in China was investigated by questionnaire and the survey data was analyzed by SPSS. Research contents included course beginning years, majors, class hours, characteristics of the course, teaching ways, the theory and practice contents, evaluation modes, selection of teaching material, teaching achievements, teachers and so on for undergraduates and graduates. Research results showed that with 20 years' development, Molecular Pharmacognosy had been offered for both undergraduate and graduate students in at least 20 colleges and universities and Molecular Pharmacognosy education in China showed good development momentum. At the same time, to promote the development of Molecular Pharmacognosy further, investment for it should be increased and practical teaching condition should be improved.
China ; Humans ; Molecular Biology ; education ; manpower ; methods ; trends ; Pharmacognosy ; education ; manpower ; methods ; trends ; Plants, Medicinal ; chemistry ; genetics ; Surveys and Questionnaires ; Teaching ; manpower ; methods ; trends

OBJECTIVETo observe the effects of bitumen fume on neurotransmitter and ultrastructure of mice brain and to investigate the toxicity of bitumen fume on nerve system of mice brain.

METHODSThe experimental mice were forced to inhale the bitumen fume at different exposure level and in different time periods. The contents of the three transmitters dopamine (DA), norepinephrine (NE), 5-hydroxytryptamine (5-HT) in mice brain were measured by the fluorescence meanwhile ultrastructure of mice brain was observed by electronic microscope. The ultrastructure of mice brain was observed under transmission electron microscopy.

RESULTSThe contents of DA, NE and 5-HT in all groups decreased with the increasing of dose and prolonging of time (after 8 week, with the increasing of exposure lever, the content of DA, NE, 5-HT was respectively 2.194, 2.190, 2.181, 2.178 microg/g and 1.148, 1.138, 1.135 and 1.407, 1.403, 1.395 microg), but the results did not show significant differences. The structure of the mitochondria changes included the swollen mitochondria, chromatin margination, pyknosis and apoptosis in neuro cells and the processes of swollen astrocyte cells.

CONCLUSIONThe bitumen fume could induce changes of the ultrastructure of mice brain and affect the contents of neurotransmitter of mice brain.

Animals ; Brain ; metabolism ; ultrastructure ; Female ; Hydrocarbons ; toxicity ; Male ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; analysis

Our previous results have demonstrated that insulin reduces myocardial ischemia/reperfusion (MI/R) injury and increases the postischemic myocardial functions via activating the cellular survival signaling, i.e., phosphatidylinositol 3-kinase (PI3-K)-Akt-endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) cascade. However, it remains largely controversial whether c-Jun NH2-terminal kinase (JNK) is involved in the effects of insulin on MI/R injury. Therefore, the aims of the present study were to investigate the role of JNK, especially the cross-talk between JNK and previously expatiated Akt signaling, in the protective effect of insulin on I/R myocardium. Isolated hearts from adult Sprague-Dawley rats were subjected to 30 min of regional ischemia and followed by 2 or 4 h of reperfusion (n=6). The hearts were pretreated with PI3-K inhibitor LY294002, or phosphorylated-JNK inhibitor SP600125, respectively, then perfused retrogradely with insulin, and the mechanical functions of hearts, including the heart rate (HR), left ventricular developed pressure (LVDP) and instantaneous first derivation of left ventricular pressure (+/-LVdp/dt(max)) were measured. At the end of reperfusion, the infarct size (IS) and apoptotic index (AI) were examined. MI/R caused significant cardiac dysfunction and myocardial apoptosis (strong TUNEL-positive staining). Compared with the control group, insulin treatment in MI/R rats exerted protective effects as evidenced by reduced myocardial IS [(28.9 +/- 2.0)% vs (45.0 +/- 4.0) %, n=6, P<0.01], inhibited cardiomyocyte apoptosis [decreased AI: (16.0 +/- 0.7) % vs (27.6 +/- 1.3) %, n=6, P<0.01] and improved recovery of cardiac systolic/diastolic function (including LVDP and +/-LVdp/dt(max)) at the end of reperfusion. Moreover, insulin resulted in 1.7-fold and 1.5-fold increases in Akt and JNK phosphorylation in I/R myocardium, respectively (n=6, P<0.05). Inhibition of Akt activation with LY294002 abolished, and inhibition of JNK activation with SP600125 enhanced the cardioprotection by insulin, respectively. And the abolishment by LY294002 could be partly converted by SP600125 pretreatment. In addition, SP600125 also decreased the Akt phosphorylation (n=6, P<0.05). These results demonstrate that insulin simultaneously activates both Akt and JNK, and the latter further increases the phosphorylation of Akt which attenuates MI/R injury and improves heart function; this cross-talk between Akt and JNK in the insulin signaling is involved in insulin-induced cardioprotective effect.
Animals ; Apoptosis ; Heart ; Insulin ; metabolism ; JNK Mitogen-Activated Protein Kinases ; MAP Kinase Signaling System ; Myocardial Infarction ; Myocardial Ischemia ; Myocardial Reperfusion Injury ; Myocardium ; Myocytes, Cardiac ; Nitric Oxide Synthase Type III ; Phosphatidylinositol 3-Kinase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Signal Transduction

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective To observe the long term effects of arthroscopic knee debridement and reconstructing operation for treating osteoarthritis in patients with Kaschin-Beck disease. Methods Thirty-one cases of patients with Kaschin-Beck disease were followed for 6 years after operation of articular clearing by arthroscope. Index of pain, symptoms of self-evaluation, range of motion, walking distance, standing test by affected leg when bending at 30° or 60° were recorded and compared with the preoperative results. Results Twenty-four cases were followed up for 6 years. Six years after operation the pain index(3.38 ± 2.87) was dramatically decreased compared to that before operation (6.88 ± 1.45, t = 5.30, P ＜ 0.05). Patients symptoms markedly improved by subjective self-evaluation was 70.83% (17/24), the effective rate was 100% (24/24). The number of cases that could stand up when leg bending at 30° or 60° were 21,18 cases, respectively, compared with that of preoperative of 14, 11 cases, respectively, the difference was statistically significant(x2 = 5.17,4.27, all P ＜ 0.05). Six years after operation the walking distance(3 cases ＜ 1 km, 11 cases 1 - 5 km and 10 cases ＞ 5 km) were greatly improved compared to the results before operation (12 cases ＜ 1 km, 9 cases 1 - 5 km and 3 cases ＞ 5 km, U = 2.88, P ＜0.05). Six years after operation the knee activity[(132.25 ± 14.52)°] remained at the same level, compared with that of preoperative [(131 .58 ± 14.68) °], the difference was not statistically significant (t = 0.16, P ＞ 0.05) .Conclusions The method of arthroscopic joint debridement to cure Kaschin-Beck disease knee osteoarthritis can significantly reduce pain, improve function and walking distance, with more stable long-term satisfactory outcome.