OBJECTIVE:To establish HPLC fingerprint for the leaves of Camptotheca acuminante in Guizhou.METHODS:HPLC method was performed.The determination was performed on Gemini-NX C18 column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 370 ran,and the column temperature maintained at 30 2.The sample size was 10 μtL.Using sorbitol as a reference,HPLC fingerprints of 14 batches of the leaves of C.acuminante were determined.The chromatographic fingerprint was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of TCM (2004 A) in terms of common peak indentification,similarity evaluation and cluster analysis.RESULTS:There were 10 common peaks in HPLC fingerprints for 14 batches of the leaves of C.acuminate.And the similarity of 13 batches of the leaves of C.acuminate was greater than 0.90,and that of another one was less than 0.90.The leaves of C.acuminate were classified into 3 groups.CONCLUSIONS:The established fingerprint can provide reference for identification and quality evaluation of the leaves of C.acuminate.

Aim To investigate the protective effect of astragaloside IV (AS-Ⅳ) on human retinal pigment epithelium injury induced by methylglyoxal (MGO), and explore its molecular mechanism.Methods The injury of ARPE-19 cells was induced by MGO and the cell viability was measured by CCK-8 method.The morphology of cell nucleus was analyzed by Hoechst 33342 staining and the cell apoptosis was analyzed by flow cytometry to detect labbled Annexin V-FITC/PI.JC-1 staining and fluorescence probe DCFH-DA were employed to evaluate the change of mitochondrial membrane potential and reactive oxygen species (ROS).The levels of SOD, MDA, caspase-9 and caspase-3 were determined by respective kits.Western blot was used to analyse the expression of Bcl-2, Bax and PARP.Results AS-Ⅳ could significantly inhibit the decrease of cell viability induced by MGO, improve the morphology of cell nucleus, reduce the ARPE-19 cell apoptosis rate and the level of ROS and MDA, and increase the activity of SOD.Furthermore, AS-Ⅳ could enhance mitochondrial membrane potential, the ratio of Bcl-2/Bax and the expression of PARP, and inhibit the activation of caspase-9 and caspase-3.Conclusion AS-Ⅳ may protect ARPE-19 cells from the injury induced by MGO by increasing the antioxidant ability of ARPE-19 cells and inhibiting cell apoptosis.

A cobalt nanoparticles modified ITO electrode (NpCo/ITO) was prepared by casting cobalt nanoparticles onto ITO electrode and the cobalt nanoparticles were synthesized by NaHB4 reduction. The electrochemical behaviors of adriamycin (ADM) on NpCo/ITO were studied. The modified ITO electrode was characterized by cyclic voltammetry (CV), scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). In a 0.01 mol L(-1) PBS (pH 8.0) buffer solution, a sensitive reduction peak of ADM was obtained. A linear relationship is held between the peak current and ADM concentration in the range of 1.0 x 10(-8) - 2.0 x 10(-6) mol L(-1) with detection of 5.0 x 10(-9) mol L(-1)- by cyclic voltammetry (CV) response. The reduction process was irreversible with adsorption at the NpCo/ITO electrode. The modified electrode showed an excellent electrocatalytic activity for the ADM electrochemical reduction.
Antineoplastic Agents ; chemistry ; pharmacology ; Clinical Laboratory Techniques ; Cobalt ; chemistry ; Doxorubicin ; chemistry ; pharmacology ; Electrochemistry ; Electrodes ; trends ; Nanoparticles ; chemistry

Aim To explore the role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in high fat-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs).Methods HUVECs were exposed to different concentrations of palmitic acid(0.1,0.2,0.4,0.8 mmol · L-1) for 24 h and different time points of 0.4 mmol · L-1 palmitic acid(0,12,24,48 h).Cell viability was measured by Cell Counting Kit 8,and the protein expression of NADPH oxidase subunits such as p22phox,p47phox,p67phox and gp91phox were determined by Western blot.The expression of reactive oxygen species (ROS) in HUVECs was detected by immunofluorescence.Results Cell proliferation rate of HUVECs stimulated by 0.4 mmol · L-1 palmitic acid for 24 h and 48 h was significantly reduced.In the next experiment,model group was accordingly set as HUVECs stimulated by 0.4 mmol · L-1 palmitic acid for 24 h.The expression of NADPH oxidase subunits such as p22phox,p47phox,p67phox and gp91phox significantly increased at 24 h and 48 h after 0.4 mmol· L-1 palmitic acid stimulation (P ＜ 0.05),and the difference.between the 24 h group and the 48 h group was not significant (P ＞ 0.05).The expression of ROS in HUVECs significantly increased at 24 h and 48 h after O.4 mmol · L-1 palmitic acid stimulation (P ＜ 0.05),and the difference between 24 h group and 48 h group was not significant (P ＜ 0.05).Compared with the model group (0.4 mmol · L-1 palmitic acid stimulation for 24 h),the NADPH oxidase inhibitor diphenyliodonium (DPI,10 μmol · L-1) pretreatment could significantly decrease the expression of ROS in vascular endothelial cells (P ＜ 0.05).Conclusion Activated NADPH oxidase might play an important role in treatment of high fat-induced oxidative stress injury in vascular endothelial cells.

OBJECTIVETo analyze the relationship between high-performance liquid chromatography (HPLC) fingerprints of the chloroform extract fractions of Peucedanum harrysmithii var. subglabrum (PHS) and its phlegm-reducing effect, in order to establish "active component group for reducing phlegm".

METHODHPLC was adopted to determine and analyze HPLC fingerprints of chloroform extract fractions of PHS. Phenol red expectorant experiment was used to observe the phlegm-reducing effect in mice. Mice were administered intragastrically with chloroform extract fractions for 6 days (1.4 g x kg(-1)), with acute bronchitis syrup as the positive control drug (12 mL x kg(-1)). The phenol red secretion in mice was determined by spectrophotometer. Then the grey relational analysis was used to study the spectrum-effect relationship.

RESULTThe phlegm-reducing effect of the chloroform extract fractions of PHS were resulted from the combined effect of all of its chemical components. Its various characteristic peaks represented different chemical components, and the order of their contributions to the phlegm-reducing effect was (number of peaks) 13 > 12 > 16 > 18 > 19 > 6 > 20 > 14 > 1 > 11 > 15 > 10 > 17 > 2 > 5 > 4 > 7 > 3 > 8 > 9, in No. 1, 3, 4, 10, 13 and 16 characteristic peaks were identified as marmesin, psoralen, xanthotoxin, Pd-Ib, pteryxin and peuformosin.

CONCLUSIONThe chloroform extract fractions of PHS show strongly phlegm-reducing effect. There may be certain relationship between their HPLC fingerprint and phlegm-reducing effect.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Ferns ; chemistry ; Mucus ; drug effects

Aim To explore the role of endoplasmic reticulum stress(ERS)signaling pathway in high fat-induced cell apoptosis in human umbilical vein endo-thelial cells(HUVECs). Methods HUVECs were ex-posed to different concentrations of palmitic acid(0. 1, 0. 2,0. 4,0. 8 mmol·L - 1 )for 24 h and different time points of 0. 4 mmol·L - 1 palmitic acid(0,12, 24,48 h). Cell viability was measured by cell count-ing kit(CCK-8),and the protein expressions of ERS signaling pathway protein such as GRP78,CHOP, PERK,IRE1,ATF6 were determined by Western blot. The level of intracellular apoptosis was detected by immunofluorescence. Results HUVECs exposed to palmitic acid at 0. 4 mmol·L - 1 for 24 h showed a de-crease in their viability and an increase in the expres-sion of ERS signaling pathway proteins (GRP78, CHOP,PERK,IRE1,ATF6)(P < 0. 05);cell ap-optotic levels significantly increased(P < 0. 05). The intracellular apoptosis levels in the vascular endothelial cells of ERS signaling pathway inhibitor 4-phenylbutyr-ic acid (4-PBA,10 mmol · L - 1 )were significantly lower than those of the PA group(P < 0. 05). Conclu-sion Activated ERS signaling pathway might play an important role in the treatment of high fat-induced cell apoptosis in vascular endothelial cells.

Objective To explore the difference of working status and attitude of nursing students of different education levels at late stage of intemship.Methods 794 nursing students of different education levels were investigated with self-made questionnaire.Resuits Nursing students of different education levels had significant difference in working status and attitude at late stage of intemship.P<0.01 or P<0.05.Undergraduate students displayed better organization and discipline practice than junior college and technical secondary nursing school students and they asked for more time off than iunior college and technical secondary nursing school students.The former tend to leave for iob hunting,while the latter for lack of working enthusiasm.Significant difference was also found in working attitude,which was mainly complaining and loss for undergraduate and junior college students and anxiety for technical secondary school students.Junior coilege and technical secondary school students preferred technical nursing,while undergraduate stedents liked technical operation.condition observation,health education,psychological care and other nursing care.Conclusions Medieal insfitutions and nursing schools should take effective strategies aimed at working status and attitude of nursing students of different education levels at late stage of internship.To ensure quality of training.mentality deviation should be rectified.leaving frequency should be controlled and the concept that health care is more important than nursing and comprehensive ability should be corrected.

OBJECTIVETo study the changes of pharmacokinetics of 6,7-dimethoxycoumarin in a rat model of alpha-naphthylisothiocyanate (ANIT)-induced experimental hepatic injury after oral administration of Yinchenhao Decoction (, YCHD) using an ultra pressure liquid chromatography (UPLC) method.

METHODSRats were divided into a normal group and a model group, after modeled by 4% ANIT (75 mg/kg) for 48 h, they were orally administrated with YCHD extract at the dose of 0.324 g/kg, and then blood was collected from their orbital sinus after different intervals. Changes in liver function were monitored by the levels of liver enzymes [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] and bilirubins [total bilirubin (TBIL), direct bilirubin (DBIL)], the concentration of 6,7-dimethoxycoumarin in plasma were measured by UPLC, and the pharmaceutical parameters were calculated with DAS2.1.1 software.

RESULTSThe concentration-time curve of both normal and modeled rats after oral administration of YCHD was obtained. Their time to maximum plasma concentration (t(max)) were both 0.25 h, the maximum concentration (C(max)) were 4.533 μg/mL and 6.885 μg/mL, and their area under concentration-time curve (AUC)(0→24h) were 16.272 and 32.981, respectively. There was a 51.88% and 100.46% increase in C(max) and AUC(0-t) (P<0.05), but there showed a 45.52% and 92.93% reduction in clearance of drug and volum of distribution (P<0.05), respectively.

CONCLUSIONSHepatic injury could significantly influence the pharmacokinetics of 6,7-dimethoxycoumarin after oral administration of YCHD, the absorption and distribution process was accelerated in liver injured rats, but the metabolism and elimination process was slowed. And this may lead to a significant accumulation of 6,7-dimethoxycoumarin in the body.

1-Naphthylisothiocyanate ; Administration, Oral ; Animals ; Chemical and Drug Induced Liver Injury ; blood ; drug therapy ; metabolism ; physiopathology ; Coumarins ; blood ; pharmacokinetics ; Disease Models, Animal ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; therapeutic use ; Liver ; drug effects ; physiopathology ; Models, Biological ; Rats ; Rats, Sprague-Dawley ; Validation Studies as Topic

Objective To investigate the effects of different dialysates on expression of protein kinase C-? (PKC?) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKC? specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKC? mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group, normal control group and fluid A+powder B+rottlerin group (P0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKC?. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

OBJECTIVETo explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.

METHODSForty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.

RESULTSThe conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.

CONCLUSIONLong-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.

Animals ; Blood-Testis Barrier ; Body Weight ; Connexin 43 ; metabolism ; Down-Regulation ; Fertilization ; Male ; Particulate Matter ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; Seminiferous Tubules ; Sperm Count ; Spermatocytes ; Testis ; metabolism ; pathology ; Vehicle Emissions ; toxicity