UPLC-MS/MS determination of tanshinone ⅡA, salvianolic acid B and ginsenoside Rg₁ in Fufang Danshen preparation in rat plasma and brain tissues / 中国中药杂志

A sensitive and specific ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method was developed for analysis of tanshinone ⅡA(TSⅡA), salvianolic acid B(SAB) and ginsenoside Rg₁ (GRg₁) in rat plasma and brain tissues. Male healthy Sprague-Dawley(SD) rats were orally given single dose of Fufang Danshen preparation (TS ⅡA 60 mg•kg⁻¹, SAB 300 mg•kg⁻¹, GRg₁ 150 mg•kg⁻¹, borneol 300 mg•kg⁻¹), and their blood samples and brain tissues were collected at different time points. The drug plasma and brain tissue concentrations of the three analytes were determined by UPLC-MS/MS method. Subsequently, the main pharmacokinetics parameters of plasma and brain tissues were calculated by using Phoenix WinNolin 6.1 software. The methodological test showed that all of analytes in both plasma and brain homogenate exhibited a good linearity within the concentration range(r>0.992 2). Their mean recoveries were between 58.86% and 112.1%. Intra-day and inter-day precisions of the investigated components exhibited RSD≤9.7%, and the accuracy(RE) ranged from -9.68% to 8.20% at all quality control levels. The results of accuracy and stability meet the requirements for biopharmaceutical analysis. For TSⅡA, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the plasma were (1.58±0.081) h, (725.4±88.20) μg•L⁻¹, (2 101.3±124.85) μg•h•L⁻¹ and (3.66±0.05) h, respectively. For SAB, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the plasma were (1.29±0.21) h, (307.9±46.75) μg•L⁻¹, (537.4±88.24) μg•h•L⁻¹ and (2.08±0.11) h, respectively. For GRg₁, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the plasma were (1.42±0.20) h, (460.38±154.60) μg•L⁻¹, (383.4±88.16) μg•h•L⁻¹ and (1.87±0.046) h, respectively. For TSⅡA, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the brain tissue were (0.75±0.22) h, (1.41±0.42) ng•g⁻¹, (4.34±2.48) ng•h•g⁻¹ and (4.00±1.90) h, respectively. For SAB, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the plasma were (1.08±0.20) h, (21.09±4.850) ng•g⁻¹, (14.83±3.160) ng•h•g⁻¹ and (0.99±0.08) h, respectively. For GRg₁, the pharmacokinetics parameters Tmax, Cmax, AUC0-t, MRTlast in the plasma were (0.50±0.16) h, (130.96±54.220) ng•g⁻¹, (136.24±34.350) ng•h•g⁻¹ and (2.87±0.33) h, respectively. The developed method was successfully applied in pharmacokinetic studies on content of TS ⅡA, SAB and GRg₁ in rat plasma and brain tissues.

Influence of G-protein β-Polypeptide 3 C825T Polymorphism on Antihypertensive Response to Telmisartan and Amlodipine in Chinese Patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>G-protein β-polypeptide 3 (GNB3) is a β subunit isoform of G-protein that plays important role in signal transduction of membrane G-protein coupled receptors (GPCRs). The GNB3 splice variant C825T (rs5443) is associated with risk for essential hypertension (EH) and efficacy of therapeutic drugs targeting GPCRs. It is unknown whether the polymorphism is associated with blood pressure (BP) response to telmisartan or amlodipine, two widely prescribed antihypertensive drugs.</p><p><b>METHODS</b>A total of 93 subjects initially diagnosed as EH were recruited and underwent a 4-week treatment with telmisartan (42 patients) or amlodipine (51 patients) monotherapy. Both baseline and after-treatment BP were measured. GNB3 C825T polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism.</p><p><b>RESULTS</b>Baseline systolic BP (SBP) and diastolic BP (DBP) were comparable among C825T genotypes in both telmisartan and amlodipine treatment groups. Patients with the CT or TT genotypes showed significantly lower body mass index (BMI) as compared with CC homozygotes in both groups (P < 0.05, respectively). GNB3 825TT homozygotes showed significantly higher after-treatment DBP and mean arterial pressure (MAP) than those carrying at least one 825C allele (P < 0.01) in the telmisartan treatment group. No difference in after-treatment SBP, DBP, and MAP levels among C825T genotypes was observed in the amlodipine treatment group. No significant difference in absolute changes in BP levels was observed among the genotypes in either treatment group.</p><p><b>CONCLUSION</b>The GNB3 C825T splice variant is associated with the DBP-lowering effect of telmisartan but not amlodipine in Chinese EH patients.</p>

Role of miR-511 in the Regulation of OATP1B1 Expression by Free Fatty Acid

MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter. We sought to evaluate the effect of miR-511 on the regulation of OATP1B1 expression by free fatty acids. When using free fatty acids to stimulate Chang liver cells, we found that the expression of miR-511 increased significantly while the expression of OATP1B1 decreased. We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay. Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells. Our study indicates that miR-511 may play an important role in the regulation of OATP1B1 expression by free fatty acids.

Effect of pitavastatin in different SLCO1B1 backgrounds on repaglinide pharmacokinetics and pharmacodynamics in healthy Chinese males

The effect of pitavastatin and SLCO1B1 genetic background on the pharmacokinetic and pharmacodynamic properties of repaglinide was investigated. In this randomized, placebo-controlled, crossover study, twelve healthy Chinese males were administered with pitavastatin 4 mg/d or the placebo for 5 d followed by repaglinide 4 mg given orally on d 5. Plasma repaglinide and glucose levels were measured by liquid chromatography-tandem mass spectrometry [LC/MS/MS] and the glucose oxidase method, respectively. Treatment with pitavastatin significantly increased the peak plasma concentration [C[max]] of repaglinide [P=0.003] in SLCO1B1[asterisk]1b homozygotes [P=0.015] and SLCO1B1[asterisk]15 carriers [P=0.031]. Treatment with pitavastatin led to a marginal increase in the area under plasma concentration-time curve from 0 h to infinity [AUC[0][rightwards arrow][infinity]] of repaglinide [P=0.091]. There was no significant difference in pharmacokinetic parameters or hypoglycemic effects of repaglinide among SLCO1B1 genotypes in either the pitavastatin or control group. Pitavastatin increased the C[max] of the plasma concentration of repaglinide in an SLCO1B1 genotype dependent manner, but had no apparent effect on the pharmacodynamics of repaglinide in healthy volunteers. The p values for this statement were not reported

Study on syndrome quantification, differentiation and classification of traditional Chinese medicine with data envelopment analysis / 中国中药杂志

To raise the syndrome sequence quantification, differentiation and classification algorithm based on data envelopment analysis for solving the modeling issue of syndrome differentiation and classification of traditional Chinese medicine (TCM). This algorithm has three steps: first, in order to obtain basic units for explaining pathogenesis, and establish a syndrome collection on this basis mechanisms of syndrome differentiation and classification were analyzed and classified according to TCM theory, mechanisms of syndrome differentiation and classification were analyzed and classified according to TCM theory; second, regularity and syndromes of corresponding prescriptions were sought according to the incidence and development progress of syndromes, and mathematical tools of data envelopment analysis were used to calculate state data of syndromes in each stage and obtain quantitative syndrome sequence; finally, syndrome sequence was taken as the measurement standard to quantify candidate syndromes and diagnostic information, and the similarity was calculated to obtain the matching degree between diagnostic information and candidate syndromes, so as to complete the syndrome differentiation and classification calculation. According to the results of model-based reasoning, the algorithm could indicate the regularity implied in prescription materials, and grasp the dynamic process of syndromes in an all-round way, and its results were verified through calculation and analysis on clinical cases. At least, it provides an idea for quantitative modeling of TCM.

Modeling and simulation activities to design sampling scheme for population pharmacokinetic study on amlodipine / 药学学报

Reasonable sampling scheme is the important basis for establishing reliable population pharmacokinetic model. It is an effective method for estimation of population pharmacokinetic parameters with sparse data to perform population pharmacokinetic analysis using the nonlinear mixed-effects models. We designed the sampling scheme for amlodipine based on D-optimal sampling strategy and Bayesian estimation method. First, optimized sample scenarios were designed using WinPOPT software according to the aim, dosage regimen and visit schedule of the clinical study protocol, and the amlodipine population model reported by Rohatagi et al. Second, we created a NONMEM-formatted dataset (n = 400) for each sample scenario via Monte Carlo simulation. Third, the estimation of amlodipine pharmacokinetic parameters (clearance (CL/F), volume (V/F) and Ka) was based on the simulation results. All modeling and simulation exercises were conducted with NONMEM version 7.2. Finally, the accuracy and precision of the estimated parameters were evaluated using the mean prediction error (MPE) and the mean absolute error (MAPE), respectively. Among the 6 schemes, schemes 6 and 3 have good accuracy and precision. MPE is 0.1% for scheme 6 and -0.6% for scheme 3, respectively. MAPE is 0.7% for both schemes. There is no significant difference in MPE and MAPE of volume among them. Therefore, we select scheme 3 as the final sample scenario because it has good accuracy and precision and less sample points. This research aims to provide scientific and effective sampling scheme for population pharmacokinetic (PK) study of amlodipine in patients with renal impairment and hypertension, provide a scientific method for an optimum design in clinical population PK/PD (pharmacodynamics) research.

Expression of PDCD5 in multiple myeloma and its relation with BCL-2 / 中南大学学报(医学版)

OBJECTIVE@#To determine the expression of apoptosis related gene PDCD5 in multiple myeloma (MM), and to analyze the relation between PDCD5 and BCL-2.@*METHODS@#The expressions of PDCD5 and BCL-2 protein and mRNA were determined by immunohistochemical staining method, flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR) method in bone marrow mononuclear cells. We also analyzed the relation between PDCD5 and BCL-2.@*RESULTS@#Immunohistochemical staining showed that PDCD5 protein positive cell percentage, staining intensity index (SII) of PDCD5 protein, BCL-2 protein positive cell percentage, and SII of BCL-2 protein were (34.75 +/- 6.49)%, (281.16 +/- 75.33), (29.97 +/- 5.57)%, and (224.94 +/- 57.72) in the MM group and (52.98 +/- 5.84)%, (462.84 +/- 39.77), (5.56 +/- 1.95)%, and (27.84 +/- 9.75) in the control group (all P < 0.05). Results of FCM showed that PDCD5 protein positive percentage and mean fluorescence intensity of PDCD5 were (78.11 +/- 21.63)% and (61.73 +/- 11.04) in the MM group and (89.46 +/- 9.98)% and (353.04 +/- 123.26) in the control group (all P < 0.05). RT-PCR showed that relative expression of PDCD5 and BCL-2 mRNA were (0.33 +/ -0.07) and (0.33 +/- 0.08) in the MM group and (0.53 +/- 0.05) and (0.12 +/- 0.02) in the control group (all P < 0.05). The positive cell percentage of PDCD5 and BCL-2 protein was negative correlation (r = -0.86, P < 0.05); the expression of PDCD5 and BCL-2 mRNA was the same status (r = -0.90, P < 0.05).@*CONCLUSION@#The expressions of PDCD5 protein and mRNA in MM patients are down-regulated, but the expressions of BCL-2 protein and mRNA are up-regulated. The mRNA and protein expression of PDCD5 and BCL-2 has negative correlation.

High-performance liquid chromatography-mass spectrometry for determining olmesartan in human plasma / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a simple and rapid HPLC-MS method for determining the contents of olmesartan in human plasma.</p><p><b>METHODS</b>Plasma were precipitated with trifluoroacetic acid, then analyzed on an HyPurity C(18) column (150 mm 2.1 mm, 5 microm). Samples at 40 degrees celsius;. The mobile phase consisted of water-methanol- acetonitrile(14:60:26) with a flow rate of 0.22 ml/min.</p><p><b>RESULTS</b>The lower limit of qualification was 25 microg/L. The calibration curve was linear over the range of 25-3200 microg/L (r=0.9998), with the intra-day and inter-day RSD less than 15%.</p><p><b>CONCLUSION</b>The method is sensitive, rapid and suitable for the study of pharmacokinetics and bioavailability of olmesartan.</p>

Expression of chemokine like factor-1 in nephridial tissue of lupus nephritis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the influence of chemokine like factor-1(CKLF-1) on the genesis and development of lupus nephritis.@*METHODS@#Immunohistochemistry stain was employed on the 34 lupus nephridial tissues and 10 nephridial tissues of control group. And light microscopy was applied to observe the percentage of CKLF-1 positive cells of renal gromerulus and tubule.@*RESULTS@#The percentages of CKLF-1 positive cells of renal gromerulus and tubule were 10.4%+/-1.5% and 48.9%+/-4.3% in the lupus nephritis group, and the percentages of control group were 4.6%+/-1.1% and 4.3%+/-0.9%. There was significant difference between 2 groups (P<0.01). And there was positive correlation between the percentage of CKLF-1 positive cells of renal gromerulus and tubule and lupus active index (r=0.74 and r=0.53, respectively, P<0.05).@*CONCLUSION@#CKLF-1 plays a role in the genesis and development of lupus nephritis.

MEF2A gene and susceptibility to coronary artery disease in the Chinese people / 中南大学学报(医学版)

OBJECTIVE@#To explore MEF2A gene and susceptibility to coronary artery disease in the Chinese.@*METHODS@#One hundred seventy-five coronary artery disease (CAD) patients and 228 normal subjects were recruited and their blood samples were amplified to detect sequences of all 11 exons of MEF2A gene by PCR. Single-strand conformational polymorphism (SSCP) analysis was used to detect the mutation. The amplified products were purified and sequenced.@*RESULTS@#The tri-nucleotide (CAG) length polymorphism in the last coding exon of MEF2A in the Chinese was revealed and 4 of the 175 (2.3%) CAD samples containing 4 prolines were due to one proline deletion in MEF2A gene. But all the 228 normal subjects contained 5 prolines. The mutation in both 175 CAD samples and 228 normal subjects was not found in other exons.@*CONCLUSION@#The deletion mutation in exon 11 in MEF2A gene may be related to CAD susceptibility in the Chinese population.

A clinical analysis of intensive gastrointestinal lavage method for treatment of 100 severe cases with stress ulcer bleeding in neurosurgery department / 中国中西医结合急救杂志

Objective To discuss the intensive gastric cavitary lavage method in treatment of stress ulcer bleeding(SUB).Methods The therapeutic effects of 100 patients with severe SUB in intensive care unit (ICU)from July 2005 to February 2008 treated with intensive gastric cavity lavage method were observed and analyzed.Results In these 100 cases,the bleeding was stopped successfully in 97 cases,the therapeutic effective rate being 97%;there were 17 cases died of complicated pulmonary infection(17%)and 3 cases of bleeding(3%),the total fatality rate being 20%.All the other cases were cured.Conclusion Intensive gastrointestinal lavage therapy is an effective method as acid inhibitor for treatment of SUB,but it can decrease the inhabitant flora in the stomach to pass through gastric-pulmonary route,thus the incidence of pulmonary infection was reduced.

Application of FTIR spectroscopy to the analysis of eleven kinds of Dendrobium / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an FTIR method for the analysis of Dendrobium.</p><p><b>METHOD</b>Using fourier transform infrared spectrometer to record the characteristic spectra of eleven samples of Dendrobium, and to compare the spectra by PCA (principal component analysis).</p><p><b>RESULT</b>The FTIR spectra of the upper part of the stem displayed significant differences between fresh and dried samples of Dendrobium. On the other hand, differences were observed in the spectra of the middle and lower parts of stems of D. guangxieuse when compared to other species.</p><p><b>CONCLUSION</b>The method of applying PCA to FTIR analysis is a rapid and dependable method for comparing samples of Dendrobium.</p>

Role of calcineurin in the progression of cardiac hypertrophy in renovascular hypertensive rats / 生理学报

The present study was to investigate the mRNA, protein expression and the activity of calcineurin in the hypertrophic heart, and to determine the effect of calcineurin inhibitor--cyclosporine A (CsA) on the regression of cardiac hypertrophy in renovascular hypertensive rats. Renovascular hypertension was induced by two kidney-one clip methods. Two months after the operation, cardiac hypertrophy was determined by histological analysis performed in some rats (2K1C-2M), then the rats were subdivided into 2 groups: (1) 3-month old two kidney-one clip group (2K1C-3M) with rats receiving 0.9% NaCl per day for one month, and (2) CsA-treated group with rats treated with CsA for one month. Sham-operated rats were used as control. The ratio of the left ventricular weight to tibial length (LVW/TL), the area of cardiac myocyte, mRNA and protein expression and the activity of calcineurin were determined. Both the LVW/TL and the cardiomyocyte area were significantly larger in 2K1C-2M and 2K1C-3M rats than in age-matched sham-operated rats. Treatment with CsA significantly attenuated the increase in the LVW/TL as well as the cardiomyocyte area. The mRNA, protein expression and the activity of calcineurin were significantly higher in 2K1C-2M and 2K1C-3M rats than those in the age-matched sham-operated rats, while the elevation of mRNA, protein expression and activity of calcineurin were significantly suppressed in the CsA-treated rats. In conclusion, calcineurin plays a role in the progression of cardiac hypertrophy in renovascular hypertensive rats. The inhibition of calcineurin can reverse cardiac hypertrophy.

Effect of herbal tongxinluo on thrombus formation and neointimalhyperplasia of the balloon injured rabbit peripheral arteries / 中国临床药理学与治疗学

Aim The effect of herbal tongxinluo on the thrombus formation and neointimal hyperplasia of the balloon injured rabbit abdominal aorta and iliac artery was observed.Methods 36 New zealand rabbits were divided randomly into 3 groups:tongxinluo group (16 rabbits);aspirin group(11 rabbits) and control group(9 rabbits). The rabbits were treated with herb tongxinluo 0.38 g?kg-1?d-1, aspirin 50 mg?kg-1?d-1 and 0.09% natrium chloride 10 ml?d-1 alternatively. The arterial injury model was made 1 week after taking the medicine and the injured segments were taken at 24 hours,1 week or 1 month after the procedure, then the samples were stained by hematoxylin and eosin. The thrombus formation and the neointimal hyperplasia of the injured arteries were observed under microscope. Results The arterial neointimal hyperplasia was seen in both abdominal aorta and iliac artery 24 hours and 7 days after the balloon injury in each group, which led to stenosis in 5%～40% of the arterial lumen, the neointimal hyperplasia was much more significant in 2 rabbits 1 month after the balloon injury. The ratio of the arteries with thrombosis in aspirin group was less than that in the other 2 groups, but the difference was not significant. The ratio of neointimal hyperplasia in tongxinluo group was significantly less than that in aspirin and control groups. Conclusion The balloon injured rabbit abdominal aorta and iliac artery appeare to be an ideal model in researching restenosis after percutaneous transluminar coronary angioplasty (PTCA) . Herbal Tongxinluo significantly inhibit the neointimal hyperplasia of the balloon injured rabbit peripheral arteries, which indicate that herbal Tongxinluo might be a promising medicine in preventing restenosis.