To establish a LC-MS/MS method for quantification of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in rats plasma and study its pharmacokinetics after administration of Mailuoning injection at a single dose to rats. Plasma samples were acidified with hydrochloric acid and extracted with ethyl acetate. The analytes were determined by LC-MS-MS using a ZOBAX SB C18 column with a mobile phase of methanol-water (containing 2 mmol x L(-1) ammonium acetic) (60:40)at a flow rate of 0.5 mL x min(-1) and detected using ESI with negative ionization mode. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 353.1/191.0 [M-H]- for chlorogenic acid, m/z 178.9/134.9 [M-H]- for caffeic acid, m/z 515.2/353.0 [M-H]-for 3,4-DCQA, m/z 193.0/133.9 [M-H]-for ferulic acid, m/z 146.9/102.9 [M-H]- for cinnamic acid and m/z 246.0/125.8 [M-H]- for tinidazole (IS). After administration of Mailuoning injection at a single dose to eight Sprague-Dawley rats, the concentrations of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in plasma were determined by LC-MS/MS method. The main pharmacokinetics parameters of measured data were caluculated by using DASver 1.0 software. The linear concentration ranges of the calibration curves for chlorogenic acid, caffeic acid, 3,4-DCQA and cinnamic acid were 2.006-1,027 microg x L(-1) (r = 0.999 6), 1.953-1,000 microg x L(-1) (r = 0.999 7), 28.51-1.459 x 10(4) microg x L(-1) (r = 0.998 9), 1.836-940.0, g x L(-1) (r = 0.997 7) and 4.780-2,447 microg x L(-1) (r = 0.998 6) respectively. The inner and inter-days relative standard deviations were both less than 5.0%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. For chlorogenic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (49.78 +/- 12.81) min, (123.55 +/- 14.82) mg x min x L(-1) and (0.004 3 +/- 0.000 5) L x min(-1), respectively. For caffeic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (36.65 +/- 10.59) min, (91.67 +/- 11.77) mg x min L(-1) and (0.005 7 +/- 0.000 7) L x min(-1), respectively. For 3,4-DCQA, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (50.08 +/- 13.78) min, (278.34 +/- 31.82) mg x min x L-1 and (0.001 6 +/- 0.000 2) L x min(-1), respectively. For ferulic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (51.39 +/- 15.52) min, (34.72 +/- 4.67) mg x min x L(-1) and (0.000 4 +/- 0.0001) L x min(-1), respectively. For cinnamic acid, the pharmacokinetic parameter t1/2, AUCo-t, and CL were (74.42 +/- 18.32) min, (34.63 +/- 4.82) mg x min x L(-1) and (0.007 7 +/- 0.001 1) L x min-', respectively. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid.
Animals ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Hydroxybenzoates ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

This study aimed to explore the impact of depression caused by chronic unpredictable mild stress (CUMS) on in vivo activity of six kinds of CYP450 isoforms in rats. According to 'Katz' method, the model of CUMS was established. Tolbutamide, chlorzoxazone, theophylline, midazolam, omeprazole and dextromethorphan were chosen as probe substrates of CYP2C6, CYP2E1, CYP1A2, CYP3A2, CYP2D1 and CYP2D2 of rats. Plasma concentration of six kinds of CYP450 in control group and model group were determined by LC-MS/MS and computed pharmacokinetic parameters. Consequently, metabolism of theophylline and chlorzoxazone accelerated significantly (P < 0.01), but tolbutamide, dextromethorphan, omeprazole and midazolam had no significant difference. The present study proved that depression caused by CUMS had strong induction to CYP1A2 and medium induction to CYP2E1.
Animals ; Chlorzoxazone ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 Enzyme System ; metabolism ; Depression ; Dextromethorphan ; metabolism ; Liver ; enzymology ; Midazolam ; metabolism ; Omeprazole ; metabolism ; Rats ; Stress, Physiological ; Tandem Mass Spectrometry ; Theophylline ; metabolism ; Tolbutamide ; metabolism

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis ; complications ; pathology ; Liver Neoplasms ; etiology ; pathology ; Male ; Middle Aged ; Young Adult

Objective - To analyze the relationship between cobalt level of post shift urine and individual exposure level of ， cobalt and its compounds in cobalt exposed workers and to explore the feasibility of using urine cobalt as a biomarker. Methods - A total of 148 occupational cobalt exposed workers from a new material company were selected as the exposed ， - - group and 44 non occupational cobalt exposed workers from the company were selected as the control group using the typical sampling method. The exposure concentration of time weighted average of cobalt and its compounds in the workplace air of the - two groups was determined by inductively coupled plasma mass spectrometry as the individual exposure level. The cobalt levels - - of pre shift and post shift urinary samples of the two groups were detected by this method. The linear relationship between the - cobalt level of post shift urine and the individual exposure level of cobalt and its compounds in the air of the workplace was Results - 3 analyzed. The individual exposure level of cobalt and its compounds in the exposed group was 1.10 131.71 μg/m with （M） 3 the median of 12.23 μg/m. No cobalt and its compounds were detected in the workplace air in the control group. The cobalt - - levels of pre shift and post shift urines in exposed group were higher than those in the control group at the same time point （M： vs ， vs ， P ） - - 1.54 0.56 μg/L 8.77 0.83 μg/L all <0.01 . The cobalt level of post shift urine was higher than that in pre shift （M： vs ，P ）， urine in the exposed group 8.77 1.54 μg/L <0.01 and it was positively correlated with the individual exposure level （ ，P ） ， of cobalt and its compounds Spearman correlation coefficient=0.86 <0.01 . After common logarithm conversion the linear regression equation of the cobalt level of post shift urine and the common logarithm of individual exposure level of cobalt and （x） ：ŷ x（ ；F ， its compounds in the exposed group was as follows = −0.178 + 0.988 coefficient of determination=0.72 =374.75 P ；t ， P ） Conclusion - <0.01 = - 19.36 <0.01 . There was a linear correlation between cobalt level of post shift urine and occupational cobalt exposure level of cobalt exposed workers. Urinary cobalt can be used as a biomarker of occupational cobalt

Objective To investigate the status of social adjustment in undergraduate nursing students, and to explore the influence of interpersonal relations and social support on social adjustment. Methods A total of 485 undergraduate nursing students in the School of Nursing of University of South China were investigated by using the Comprehensive Diagnostic Scale for Interpersonal Relations, Social Support Rating Scale and Chinese College Students' Adaptability Scale with stratified cluster sampling method. Results The total scores of social adjustment of undergraduate nursing students were (195.37 ± 26.63) points. The total scores of interpersonal relations were (9.18 ± 5.81) points. The total scores of social support were (36.72 ± 5.39) points. Social support and social adjustment were positively correlated (r=0.274, P<0.01), but interpersonal relations and social adjustment were negatively correlated (r=-0.490, P<0.01). The main influencing factors of social adjustment in a descending order included interpersonal dating(β=-0.390, P=0.000), interpersonal conversation(β=-0.146, P =0.024) and subjective support(β=0.133, P =0.005). Conclusions The scores of the social adjustment, social support and interpersonal relations of undergraduate nursing students are all in the middle level. The main influencing factors of social adjustment include interpersonal dating, interpersonal conversation and subjective support, particulally interpersonal dating. Educators can improve their social adjustment through improving interpersonal dating, interpersonal conversation and subjective support .

The aim of the present study was to study the effect of β-estradiol (β-E(2)) on the large-conductance Ca(2+)-activated potassium (BK(Ca)) channel in mesenteric artery smooth muscle cells (SMCs). The mesenteric arteries were obtained from post-menopause female patients with abdominal surgery, and the SMCs were isolated from the arteries using an enzymatic disassociation. According to the sources, the SMCs were divided into non-hypertension (NH) and essential hypertension (EH) groups. Single channel patch clamp technique was used to investigate the effect of β-E(2) and ICI 182780 (a specific blocker of estrogen receptor) on BK(Ca) in the SMCs. The results showed the opening of BK(Ca) in the SMCs was voltage and calcium dependent, and could be blocked by IbTX. β-E(2) (100 μmol/L) significantly increased open probability (Po) of BK(Ca) in both NH and EH groups. After β-E(2) treatment, NH group showed higher Po of BK(Ca) compared with EH group. ICI 182780 could inhibit the activating effect of β-E(2) on BK(Ca) in no matter NH or EH groups. These results suggest β-E(2) activates BK(Ca) in mesenteric artery SMCs from post-menopause women via estrogen receptor, but hypertension may decline the activating effect of β-E(2) on BK(Ca).
Aged ; Estradiol ; analogs & derivatives ; pharmacology ; Female ; Humans ; Hypertension ; physiopathology ; Large-Conductance Calcium-Activated Potassium Channels ; agonists ; metabolism ; physiology ; Mesenteric Arteries ; metabolism ; physiology ; Middle Aged ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Patch-Clamp Techniques ; Postmenopause ; physiology ; Receptors, Estrogen ; antagonists & inhibitors

Objective To investigate the clinical significance of 5 kinds of inflammatory factors (MCP 1,IL 1β,IL-18,HMGB1,and IL-10) in severe EV71 hand-foot-and-mouth disease (HFMD).Methods Hospitalized children who were diagnosed with severe EV71 HFMD in a hospital in March August,2014 were as HFMD group,healthy children who underwent physical examination in outpatient department of the same hospital during the sameperiod were as control group,changes in expression levels of peripheral blood MCP-1,IL-1β,IL-18,HMGB1,and IL-10 of both groups were dynamically observed,clinical data of HFMD group were collected.Results There were 102 children in HFMD group,the average age was (2.18 ± 0.91) years old,80.39％ of whom were ≤3 years old;there were 77,16,and 9 cases in HFMD group at stage 2,3,and 4 respectively at admission.77,52,21,and 88 cases went through stage 2,3,4,and 5 respectively.Expression levels of 5 kinds of inflammatory factors at stage 2,3,and 4 in HFMD group were compared respectively with control group,differences were all statistically significant(all P＜0.05);expression levels of IL-10 at stage 3 and 4 in HFMD group were not significantly different (P＞0.05).In HFMD group,the expression levels of HMGB1 of stage 2,3 progression groups were both higher than recovery group(both P＜0.05).The expression levels of 5 kinds of inflammatory factors in the death group and survival group at admission were all significantly different (all P＜0.05).Conclusion The expression levels of MCP 1,HMGB1,IL-1β,IL-10,and IL-18 are closely related to the severity of HFMD,and has certain clinical significance for the prognosis of children.HMBG1 has certain predictive value in the prognosis of HFMD.

Objective To investigate the expression and clinical significance of peripheral blood CD4~+, CD25~+ and CD4~+CD25~+ T subpopulations in patients with systemic lupus erythematosis.Methods The per- centage and fluorescence intensities of peripheral blood CD4~+,CD25~+ and CD4~+CD25~+ subpopulations from 34 SLE and 18 normal controls were measured with flow cytometry assay,then the correlation with clincal data was analyzed.The CD25~+ cells were defined as the CD25~(high) cells if their fluorescence intensity was higher than 10. Results The percentage of CD4~+CD25~+,CD4~+CD25~(high) T lymphocytes in active SLE patients[(4.80?1.21)% and (0.25?0.10)%]was lower than that in normal controls[(8.92?3.21)% and(0.44?0.22)% and non-active SLE patients(11.28?2.09)% and(0.59?0.34)%](P＜0.05).However,as for the CD25~+ cells in the CD4~+ T cells,there was no difference between SLE patients and normal control group.Peripheral blood CD4~+CD25~+,CD4~+CD25~(high) cells in SLE were reversely correlated with SLEDAI(r=-0.74,P=0.004 and r=-0.614,P=0.026),but not with others such as complements,ANA titers etc.Peripheral blood CD4~+ and CD25~+ lymphocytes in active SLE pa- tients were also lower than those in normal controls[(23?7)vs(34?7)and(7.4?1.8)vs(13.9?3.4),P＜0.05]. CD25 fluorescence intensities were higher in the SLE patients those in the normal controls,but CD4 fluores- cence intensities were not.Conclusion CD4~+CD25~+ may play a role in the pathogenesis of SLE.

Background: Extensive studies have confirmed the efficacy of taxanes in combination with anthracycline-basedchemotherapy on breast cancer. However, few studies have assessed the efficacy of weekly taxane–anthracyclineregimens on locally advanced breast cancer. This study was to compare the efficacy and safety of a weekly taxane–anthracycline regimen with those of tri-weekly anthracycline-based regimen in patients with locally advanced breastcancer.Methods: Patients with locally advanced breast cancer were randomized to receive 4–6 cycles of neoadjuvant chemotherapywith tri-weekly 5-fluorouracil–epirubicin–cyclophosphamide (FEC) regimen or weekly paclitaxel–epirubicin(PE) regimen. The primary endpoint was the pathologic complete response (pCR) rate. Other endpoints included theclinical tumor response, breast-conserving surgery rate, and adverse events.Results: Between March 2010 and September 2013, 293 patients were randomized to the FEC (n = 151) and PE(n = 142) arms. The overall clinical response rate was significantly higher in the PE arm than in the FEC arm (76.06% vs.59.95%, P = 0.001). Consistently, the post-chemotherapy pathologic T and N stages were significantly lower in the PEarm than in the FEC arm (P < 0.001). However, the pCR rate was similar in the two arms (10.61% vs. 12.31%, P = 0.665).Overall, 36 (27.27%) patients in the FEC arm and 6 (35.28%) in the PE arm were qualified for breast-conserving surgery.Most adverse events were comparable in both arms, with more severe neutropenia in the PE arm than in the FEC arm(11.97% vs. 5.96%, P = 0.031).Conclusions: In patients with locally advanced breast cancer, weekly PE was not superior to FEC in terms of pCR.However, weekly PE has a higher response rate and superior down-staging effects. On this account, the PE regimenmay be considered an alternative option for locally advanced breast cancer. Long-term follow-up data are needed toconfirm the efficacy of this regimen on locally advanced breast cancer.

OBJECTIVETo investigate the dynamic correlation between pre-S1 antigen, pre-S2 antigen and HBV DNA in the serum of chronic hepatitis B (CHB) patients undergoing nucleoside analogue therapy.

METHODS12 CHB patients with transient virological response after lamivudine treatment, and 20 patients treated with adefovir for 5 years were recruited in this study. Serum samples were collected at four time points when HBV DNA fluctuated sharply during lamivudine treatment, and at 0, 8, 12, 28, 52, 104, 156, 208, 260 weeks following adefovir treatment. HBV DNA was quantified by real-time PCR, pre-S1 and pre-S2 antigens were detected by ELISA.

RESULTSThe titers of pre-S1 and pre-S2 antigens were not correlated with the HBV DNA level in the serum of lamivudine treated patients. Only in one case of the adfovir treated patients, the decrease of pre-S1 and pre-S2 antigens was in parallel with the decrease of HBV DNA. Linear regression analysis indicated that neither pre-S1 antigen nor pre-S2 antigen was correlated with HBV DNA in the serum of lamivudine or adfovir treated patients (P more than 0.05).

CONCLUSIONOur results indicate that the titers of pre-S1 and pre-S2 antigens are not correlated with the serum HBV DNA in CHB patients undergoing nucleoside analogue therapy. Neither pre-S1 nor pre-S2 is a good predictor for the outcome of nucleoside analogue treatment.

DNA, Viral ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Real-Time Polymerase Chain Reaction