Objective:To develop an LC-MS/ MS method for the analysis of 17 memory-enhancing drugs illegally added in tradi-tional Chinese medicines(TCM)and health foods. Methods:The method involved liquid chromatography-tandem mass spectrometry (LC-MS/ MS),and the separation was conducted on an Eclipse XDB C18 column(100 mm × 3. 0 mm,1. 8 μm). The 17 memory-en-hancing drugs illegally added in traditional Chinese medicines and health foods were analyzed by LC-MS/ MS with electrospray ioniza-tion(ESI)and multiple reactions monitoring(MRM). Results:A fast LC-MS/ MS method for the simultaneous screening of 17 illegal-ly added chemical components was described with the LOD below 10 ng·ml - 1 . Conclusion:The method is selective and sensitive in the detection of memory-enhancing drugs illegally added in traditional Chinese medicines and health foods.

Objective To investigate the therapeutic efficacies for pure galactoma by injecting absolute ethanol compared with the pure pouch cavity puncture separately pulls out the fluid law.Methods Patients of pure galactoma examined by ultrasonography were divided into two groups from March of 2005 to February of 2007 in mammary gland special clinic of our hospital.Absolute ethanol jet method and the pure pouch cavity puncture separately pulls out the fluid law were given respectively and the effect was evaluated within one year after operation.Results Of 89 cases pure galactoma using absolute ethanol jet method,71cases(79.78%)were convalesced,15cases(16.85%)were effective and 3 cases(3.37%)were noneffective.The effectiveness was 96.63 %.Of 86 cases using pure pouch cavity puncture separately pulls out the fluid law,44cases(51.16%)were convalesced,33 cases(38.37%)were effective and 9 cases(10.47%)were noneffective.The effectiveness was 89.53%.The cured rates of two groups had significant difference through chi-square test,?2=15.38,P

Objective:To study the feasibility of the determination of ephedrine hydrochloride in Tongxuan Lifei pills by acceler-ated solvent extraction ( ASE) combined with QuEChERS purification, and compare the results with those of the extraction method in Chinese Pharmacopoeia. Methods: The extraction efficiency of ephedrine hydrochloride in Tongxuan Lifei pills determined by HPLC was used as the evaluation index, and the operation parameters of ASE were optimized by orthogonal experiments. Results: The opti-mum conditions of ASE for the determination of ephedrine hydrochloride in Tongxuan Lifei pills were as follows:the samples were defat-ted by n-hexane ( the extraction temperature was 80℃, the static extraction time was 5 min for one cycle, and the flush volume was 100%. ) ,and then methanol was used as the extraction solvent for the extraction of ephedrine hydrochloride ( the extraction temperature was 80℃,the static extraction time was 8 min for three cycles) ,in the end, the impurities were purified by PSA purifying agent. Using the optimized ASE method to extract ephedrine hydrochloride in Tongxuan Lifei pills, the extraction time was reduced to 40 minutes with less interference, and compared with that of the method in Chinese Pharmacopoeia, the determination of the relative standard devi-ation was less than 5%. Conclusion:The ASE technique combined with QuEChERS purification is simple, quick and effective,and it can be used as the pretreatment method for the determination of ephedrine hydrochloride in Tongxuan Lifei pills.

Objective To investigate the clinicopathological features and risk factors of lymph node metastasis of gastrointestinal neuroendocrine neoplasms (GI-NENs).Methods The retrospective case-control study was conducted.The clinicopathological data of 467 patients with GI-NENs who were admitted to the Fourth Hospital of Hebei Medical University from January 2006 to December 2015 were collected.Observation indicators:(1) occurrence sites and pathological classification of GI-NENs;(2) pathological characteristics of surgical specimens of GI-NENs;(3) univariate analysis and multivariate analysis affecting lymph node metastasis of GI-NENs:sex,age,tumor location,tumor diameter,pathological classification,pathological stage and tumor invasive depth.The univariate analysis and multivariate analysis were respectively done using the chi-square test and Logistic regression model.Results (1) Occurrence sites and pathological classification of GI-NENs:of 467 patients with GI-NENs,tumors of 304,15,7,14 and 127 patients were located at stomach,duodenum,small intestine,colon and rectum,respectively.Tumor diameter was 0.3-12.0 cm,with an average diameter of 2.2 cm.Of 467 patients with GI-NENs,G1 and G2 of neuroendocrine tumors (NETs),G3 of neumendocfine carcinomas (NECs) and mixed adenoneuroendocfine carcinomas (MANECs) were respectively detected in 209,64,146 and 48 patients.Lymph node metastasis rate of GI-NENs was 31.48％ (147/467).(2) Pathological characteristics of surgical specimens of GI-NENs:NETs were high-differentiated NENs.Ceils of NETs were solid and nest-,trabeculum-and tubular-shaped,and consisted of small or medium cells,with moderate amount or massive cytoplasms,round or oval nucleus,particle-shaped chromatin,unobvious nucleolus and positive endocrine markers.There were abundant of small blood vessels and surrounding fibrous stroma in peripheral tumor cell nests.NECs were low-differentiated NENs and included small cell carcinoma and large cell NEC.Cells of small cell carcinoma were small round or oval and looked similar to lymphocytes,with few amount cytoplasms,fine granularshaped or hyperchromatic nucleus and common mitosis figures.Cells of large cell NEC were large and greater than 3 lymphocytes,arrayed in organoid-or chrysanthemum-shape,with massive cytoplasms,coarse particle-shaped chromatin,obvious nucleus,clear mitosis figures and large laminar-shaped necrosis.There were different positive expressions of endocrine markers between small cell carcinoma and large cell NEC.MANECs had the characteristics of glandular cavity formation of traditional adenocarcinoma and NENs.Results of immunohistochemical staining in 467 patients showed that Ki-67 of 467 patients was positive;CD56 in 379 of 428 with CD56 test was positive;synaptophysin (Syn) in 416 of 422 with Syn test was positive;cytokeratin (CK) in 354 of 396 with CK test was positive;chromogranin (CgA)in 264 of 388 with CgA test was positive;neuron specific enolase (NSE) in 287 of 346 with NSE test was positive.(3) Univariate analysis and multivariate analysis affecting lymph node metastasis of GI-NENs:results of univariate analysis showed that sex,tumor location,tumor diameter,pathological classification,pathological satge and tumor invasive depth were related factors affecting lymph node metastasis of patients with GI-NENs (X2 =20.654,18.182,26.788,184.709,163.738,195.391,P＜ 0.05).Results of multivariate analysis showed that pathological classification and pathological stage were independent influenced factors affecting lymph node metastasis of patients with GI-NENs (HR =2.129,7.171,95％ confidence interval:1.273-3.561,-2.327-22.098,P＜0.05).Conclusions GI-NENs are mostly located on the stomach and rectum.Results of immunohistochenical staining could help diagnosis of GI-NENs.Pathological classification and pathological stage are independent influenced factors affecting lymph node metastasis of patients with GI-NENs.

Objective To observe and evaluate the location of the three compartments of the pelvic floor organ and levator hiatus area in the postpartum women by transperineal real time three-dimensional ultrasound.Methods Sixty eases of maternal with the matching age and body mass index (BMI;postpartum group) and 35 non-fertile women (control group)were selected.The distance between the lower edge and the reference line was measured by the transperineal ultrasound at rest and Valsalva movement in the lowest point of bladder,cervix and rectum.Meanwhile,the area of levator hiatus under Valsalva action was measured by real time 3-dimensional ultrasound.The differences were compared between two groups.Results In the postpartum group,the lowest point of the bladder and cervix was lower than the control group at rest and Valsalva movemen (both P＜0.05).Rectum location in the rest state difference between the two groups had no statistical significance (t=0.405,P =0.398).The area of the levator hiatus of the postpartum group was significantly larger than that of the control group at the rest and Valsalva movement (t=11.253,9.625,P＜0.001).Conclusion Transperineal real time 3-dimensional ultrasound can dynamically observe the position,movement of pelvic floor organ and the area of levator hiatusin of the female.To evaluate the pregnancy and childbirth have obvious effect on the pelvic floor support tissues.

Objective:To design the small ubiquitin modification-fibroblast growth factor receptor 4 (SUMO-FGFR4) fusion gene and construct the expression vector pET22b-SUMO-FGFR4, to optimize the expression conditions. Methods:The SUMO-FGFR4 fusion gene was obtained by Overlap PCR and was connected to pET22b;the recombinant expression vector pET22b-SUMO-FGFR4 was obtained. The influence of lactose concentration, induction time,induction temperature,induction point and adding mode of lactose in the expression levels was observed,and the best induction condition was determined; then the solubility of recombinant protein was analyzed.Results:The SUMO-FGFR4 fusion protein was highly expressed,the molecular weight of the fusion protein was about 40 000 and it could bind with FGFR4 specific antibody.When the lactose concentration was 1.0 g·L-1 ,the induction time was 3 h,the induction temperature was 37℃,the value of A (600)was 0.8,the expression level was highest;but adding mode of lactose had no remarkable effect on the protein expression.The expression level of recombinant protein induced by lactose was higher than IPTG.SUMO-FGFR4 protein existed in a form of inclusion body.Conclusion:The SUMO-FGFR4 fusion protein is expressed successfully in this study while lactose is used as inducer and the best expression conditions are confirmed.

AIM: To elucidate the mechanism of arrhythmia in healed myocardial infarction (HMI), and to investigate the changes of action potential duration (APD),transient outward potassium current (I to ), delayed rectifier potassium current (I K) and inward rectifier potassium current (I K1 ) of left ventricular myocytes in noninfarcted zone of HMI. METHODS: 12 rabbits were randomly assigned in two groups: HMI group (thoracotomy and ligation of the circumflex coronary); sham-operated group (thoracotomy but no conorary ligation). 3 months after operation, whole cell patch clamp technique was used to record APD, I to , I K and I K1 of ventricular myocytes in non-infarcted zone. RESULTS: Membrane capacitance was larger in HMI group than that in sham-operated group. Action potential duration was lengthened significantly in HMI group and early after depolarization (EAD) appeared in HMI group. The densities of I to , I K,tail and I K1 were reduced significantly in HMI group (P

To elucidate the mechanism of arrhythmia in healed myocardial infarction (HMI), the changes of action potential duration (APD), transient outward potassium current (Ito), delayed rectifier potassium current (IK) and inward rectifier potassium current (IK1) of left ventricular myocytes in non-infarcted zone of HMI were investigated. Rabbits were randomly assigned into two groups: HMI group, in which animals were subjected to thoracotomy and ligation of the circumflex coronary and sham-operated group, in which rabbits underwent thoracotomy but no conorary ligation. 3 months after the operation, the whole myocyte patch clamp technique was used to record APD, Ito, IK, and IK1 of ventricular myocytes in non-infarcted zone. Our results showed that the membrane capacitance was larger in HMI group than in sham-operated group. Action potential duration was significantly lengthened in HMI group and early afterdepolarization (EAD) appeared in HMI group. The densities of Ito, I(K, tail), and IK1 were reduced significantly in HMI group, from 6.72 +/- 0.42 pA/pF, 1.54 +/- 0.13 pA/pF and 25.6 +/- 2.6 pA/pF in sham-operated group to 4.03 +/- 0.33 pA/pF, 1.14 +/- 0.11 pA/pF and 17.6 +/- 2.3 pA/pF, respectively. It is concluded that the reduced densities of Ito, I(K, tail) and IK1 in ventricular myocytes of non-infarcted zone in HMI were responsible for the prolongation of APD and the presentation of EAD which played important roles in the development of malignant arrhythmia in HMI.
Action Potentials ; Arrhythmia/*etiology ; Heart Ventricles/metabolism ; Myocardial Infarction/complications ; Myocardial Infarction/metabolism ; Myocardial Infarction/*pathology ; Myocytes, Cardiac/*cytology ; Patch-Clamp Techniques ; Potassium Channels/*metabolism

Objective To investigate clinical effect of fluvastatin in the treatment of early diabetic nephropa-thy,to provide a reference for clinical treatment.Methods 90 patients with diabetic nephropathy were selected,the patients were divided into observation group(50 cases)and control group(40 cases).Conventional hypoglycemic ther-apy used in the control group,and valsartan treatment also used.The observation group received fluvastatin on the basis of treatment of control group.The efficacy,urinary albumin excretion rate,inflammatory markers,serum creatinine and other indicators and adverse reactions were compared.Results The effective rate of the observation group was 90.00%,which was significantly higher than 75.00% of the control group,the difference was statistically significant (χ2 =4.325,P <0.05).After treatment,the BUN,UAER,TC,TG,LDL of the observation group were (6.54 ± 1.24)mmol/L,(40.43 ±4.21)μg/min,(3.81 ±0.47)mmol/L,(2.51 ±0.34)mmol/L,(2.41 ±0.64)mmol/L, the improvement was better than the control group,the differences were statistically significant (t =5.547,5.225, 5.457,4.957,5.339,all P <0.05).After treatment,the CRP,IL -6,IL -18,TNF -αin the observation group and control group were significantly improved compared with before treatment,the differences were statistically significant (P <0.05 ).After treatment,the CRP,IL -6,IL -18,TNF -αlevels of the observation group were (4.14 ± 0.87)mg/L,(88.17 ±8.54)pg/mL,(139.64 ±9.48)ng/L,(40.17 ±5.22)ng/L,the improvement was better than the control group,the differences were statistically significant (t =6.914,6.357,5.847,7.054,all P <0.05 ). Conclusion Fluvastatin in the treatment of early diabetic nephropathy has good effect,which will help to improve inflammatory cytokines and proteinuria and protect renal function,it is worthy of clinical application.

Larotaxel, a new taxane compound prepared by partial synthesis from 10-deacetyl baccatin Ⅲ, is active against tumors. In this research, a selective LC–MS method was developed and validated for the study of degradation kinetics of larotaxel, which was carried out in aqueous solutions with different pH (1.5, 3.0, 5.0, 6.5, 7.4, 9.0, 10 and 11.0) and temperature (0, 25, 37 and 45 °C). The linear range was 0.5–25μg/mL, the intra-and inter-day precisions were less than 7.0%, and accuracy ranged from 97.4–104.5% for each analyte. The observed rate obtained by measuring the remaining intact larotaxel was shown to follow first-order kinetics. The activation energies for degradation were 126.7 and 87.01 kJ/mol at pH 1.5 and 11, respectively. Although larotaxel was stable in pH 5, 6.5 and 7.4 buffers at 37 °C for 24 h during our study, increasing or decreasing the pH of the solutions would decrease its stabilities. Moreover, three main degradation products in alkaline condition were separated by HPLC and identified by Q–TOF–MS. The three degradation products were confirmed as 10-deacetyl larotaxel, 7, 8-cyclopropyl baccatin Ⅲ and 10-deacetyl-7, 8-cyclopropyl baccatin Ⅲ.