Objective • To investigate the effect of content changes of neurotransmitters on multiple physiological and pathophysiological processes in mammals, and explore simultaneous sensitive quantification of these metabolites. Methods • An efficient and sensitive method for simultaneous quantification of 20 amino neurotransmitters was achieved through pre-column N-ethylation of neurotransmitters with acetaldehyde and NaBH3CN followed with quantitative analysis using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). And NaBH3CN and NaBD3CN were used as a pair of stable isotope labeling reagents together with natural isotopic standards to achieve accurate quantification. Results • The analysis showed all 20 neurotransmitters had excellent linearity with R2 over 0.991 0 and sensitivity with the limits of detection (LOD) in the range of 0.5-10 fmol except for histidine with an LOD of 36.5 fmol. The result also showed good robustness with the interday and intraday RSD (relative standard deviation) below 15%. The method was further applied to human urine, serum and saliva samples for validation, in which 14, 10 and 8 neurotransmitters were detected, respectively. Conclusion • The method is of high sensitivity and precision. Compared to previous ones, this method is of great applicability in different matrices, with mild reaction and straightforward procedures. Besides, the reaction reagents are easy to be removed by simple quenching thus avoiding interference in the subsequent analysis.

To investigate the pharmacokinetic characteristics and absolute bioavailability of ginkgolide A (GA), ginkgolide B (GB) and bilobalide (BB) in rats. In this experiment, a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was established to determine the plasma concentrations of GA, GB and BB in rats after rats were administrated with the three drugs through ig and iv respectively. The main pharmacokinetic parameters and absolute bioavailability of three ginkgolide compounds were obtained by using pharmacokinetic software DAS 2. 0. After the inject of GA, GB and BB, the results showed Cmax at (513.9 ± 116.9), (701.3 ± 76.0), (5,255.6 ± 476.8) µg · L(-1) and AUC0.24h of (960.9 ± 268.5), (779.5 ± 140.6), (7,409.3 ± 1,181.1) µg · h · L(-1), respectively; after the oral administration, the results showed Cmax at (522.9 ± 39.9), (146.8 ± 31.6), (2,711.9 ± 588.9) µg · L(-1) and AUC0-24 h of (1,760.4 ± 300.7), (636.6 ± 180.3), (16,651.4 ± 1,306.5) µg · h · L(-1), respectively. The absolute bioavailability of GA, GB and BB in rats was (61.1 ± 10.4)%, (27.2 ± 7.7)%, (56.2 ± 4.4)%, respectively. The method established in this experiment has a good specificity and sensitivity and so can be used to study the pharmacokinetics and absolute bioavailability of GA, GB and BB in rats.
Animals ; Biological Availability ; Chromatography, High Pressure Liquid ; Cyclopentanes ; pharmacokinetics ; Furans ; pharmacokinetics ; Ginkgolides ; pharmacokinetics ; Lactones ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid ; methods ; Ginkgolides ; analysis ; Injections ; Limit of Detection ; Salicylates ; analysis ; Tandem Mass Spectrometry ; methods

OBJECTIVETo study the fractional anisotropy (FA) and the architecture of the optic radiation fiber tracts of normal adults with magnetic resonance (MR) diffusion tensor imaging (DTI).

METHODSDiffusion tensor images were obtained from 30 healthy volunteers without any cerebral abnormalities on conventional MRI. FA and the mean diffusivity (MD) of the optic radiation were measured in the directional encoded color (DEC) maps. The architecture of the optic radiation fiber tracts were displayed with the software of diffusion tensor fiber tracking.

RESULTSIn all subjects, the optic radiation could be readily identified in the DEC maps. The FA value was 0.509-/+0.029 in the left and 0.502-/+0.026 in the right, with the MD value of (0.763-/+0.050) x10(-3) and 0.748-/+0.052)x10(-3) mm2/s, respectively. No significant differences were found in the FA or MD value of the bilateral optic radiation (P>0.05). Diffusion tensor tractography (DTT) demonstrated that the 3 bundles of the optic radiation fibers were located in the lateral sagittal stratum, passing from the lateral geniculate body of the thalamus to the primary visual cortex. The dorsal and lateral bundles passed posteriorly to the superior bank of the calcarine cortex, while the ventral bundle passed anteriorly before making a sharp turn, known as the Meyer loop, and subsequently coursed posteriorly to terminate in the inferior margin of the calcarine cortex, which was consistent with the results of classic anatomical studies.

CONCLUSIONAs a novel method to study the relationship between visual function and optic pathway, DTI and DTT can show the FA and architecture of the optic radiation.

Adult ; Anisotropy ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Geniculate Bodies ; anatomy & histology ; Humans ; Male ; Middle Aged ; Models, Anatomic ; Occipital Lobe ; anatomy & histology ; Optic Nerve ; anatomy & histology ; Visual Pathways ; anatomy & histology ; Young Adult

To investigate the anti-cardiac hypertrophic mechanism of statins, thirty-eight male Wistar rats were randomly allocated to four groups. Rats in model group received nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA) 15 mg/(kg.d) by peritoneal injection. Rats in simvastatin treatment groups were given simultaneously L-NNA as those in model group and simvastatin 5 or 30 mg/(kg.d) intragastrically respectively. Rats in control group received the same volume of normal sodium. Left ventricular function, left ventricular mass index (LVMI), the content of brain natriuretic peptide (BNP) in plasma and myocardium, myocardial hydroxyproline and heme oxygenase activity were determined after 6 weeks. The results showed that rats in model group developed significant cardiac hypertrophy associated with reduced left ventricular function compared with the control group. However, compared with the model group, L-NNA-induced cardiac hypertrophy of rats was significantly relieved in simvastatin treatment groups, associated with improved left ventricular function, decreased LVMI, lower BNP levels in plasma and myocardium, lower content of myocardial hydroxyproline, and increased myocardial heme oxygenase (HO) activity. In cultured rat neonatal cardiomyocytes, simvastatin (30 or 100 mumol/L) significantly increased heme oxygenase-1 (HO-1) mRNA expression, HO activity as well as the production of CO in cardiomyocytes. Cultured with zinc protoporphyrin, a HO inhibitor, or simvastatin alone did not change [(3)H]leucine uptake of cardiomyocytes. However, cocultured with simvastatin significantly inhibited the cardiomyocyte [(3)H]leucine uptake induced by angiotensin II in a concentration-dependent manner. Cotreatment with zinc protoporphyrin significantly abolished the suppressive effect of simvastatin on cardiomyocyte [(3)H]leucine uptake. These data suggest that the activation of HO-1/CO pathway may be one of the important mechanisms by which statins inhibit cardiac hypertrophy caused by hypertension.
Angiotensins ; antagonists & inhibitors ; pharmacology ; Animals ; Carbon Monoxide ; metabolism ; Cardiomegaly ; etiology ; prevention & control ; Cell Enlargement ; drug effects ; Heme Oxygenase-1 ; metabolism ; Hypertension ; complications ; drug therapy ; Male ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Simvastatin ; pharmacology ; therapeutic use

OBJECTIVETo perform a Meta-analysis on peginterferon with interferon in treatment of HIV patients coinfected with refractory genotype HCV.

METHODSA literature search of Medline was conducted to identify eligible randomized controlled trials. Meta analysis was conducted to evaluate peginterferon and interferon in treatment of coinfected HCV genotype 1 or 4 in HIV patients.

RESULTSix trials of 88 matched the selection criteria. Total 1,131 patients with coinfection of HCV genotype 1 or 4 and HIV were included. Sustain viral response was higher in patients treated with peginterferon plus ribavirin compared with that of interferon plus ribavirin (26 % compared with 8 %) or peginterferon alone (26 % compared with 13 %). Severe adverse effects and withdrawal rates were similar for patients treated with peginterferon and patients treated with interferon.

CONCLUSIONPeginterferon plus ribavirin in treatment of patients with coinfection of genotype 1 or 4 HCV and HIV can achieve higher sustain viral response and the likelihoods of serious adverse effects and withdrawal rates are similar to other therapies.

Adult ; Antiviral Agents ; administration & dosage ; Drug Therapy, Combination ; Female ; Genotype ; HIV Infections ; complications ; drug therapy ; immunology ; Hepacivirus ; classification ; genetics ; Hepatitis C, Chronic ; complications ; drug therapy ; virology ; Humans ; Interferon-alpha ; administration & dosage ; Male ; Polyethylene Glycols ; administration & dosage ; Randomized Controlled Trials as Topic ; Recombinant Proteins ; Ribavirin ; administration & dosage

Objective To study the effect of OK-432 in combination with IL-2 on Lewis lung cancer (LLC) growth and bcl-2 expression in C57BL/6 mice. Methods Forty C57BL/6 mice bearing LLC were divided into 4 groups and injected with 0.9% NaCl solution, OK-432, IL-2 and OK-432/IL-2. The inhibition of tumor growth after the drug administration and the expression of bcl-2 in paraffin-embedded primary tumor tissues were observed with SABC. Results The growth of LLC in the group with combined treatment was significantly inhibited. ( with inhibition rate of 60%, P < 0.05). The expression of bcl-2 in this group was significantly lowered compared with control group (P < 0.05). Conclusions OK-432 and IL-2 has distinct synergetic anti-tumor effect, possibly by attacking the tumor cells directly or indirectly, and the improved interaction of cells due to inhibition of bcl-2 gene expression and induction of tumor cell apoptosis may also contribute to this effect.

Objective To study the effect of OK-432 in combination with IL-2 on Lewis lung cancer (LLC) growth and bcl-2 expression in C57BL/6 mice. Methods Forty C57BL/6 mice bearing LLC were divided into 4 groups and injected with 0.9% NaCl solution, OK-432, IL-2 and OK-432/IL-2. The inhibition of tumor growth after the drug administration and the expression of bcl-2 in paraffin-embedded primary tumor tissues were observed with SABC. Results The growth of LLC in the group with combined treatment was significantly inhibited. ( with inhibition rate of 60%, P < 0.05). The expression of bcl-2 in this group was significantly lowered compared with control group (P < 0.05). Conclusions OK-432 and IL-2 has distinct synergetic anti-tumor effect, possibly by attacking the tumor cells directly or indirectly, and the improved interaction of cells due to inhibition of bcl-2 gene expression and induction of tumor cell apoptosis may also contribute to this effect.

Objective To explore the clinical value of double-balloon enterocopy (DBE) in diagnosis of small intestinal diseases. Methods The clinical and endoscope image data of 231 patients with suspected small bowel disease who underwent DBE from January 2008 to May 2016 were analyzed. Result 231 patients received 257 times of DBE examination, 112 of them were performed by oral and 93 by anal route, 26 patients were underwent by both approaches. The detection rate of intestine diseases was 64.9% (150/231), include 33 cases (14.3%) of nonspecific enteritis, 27 cases (11.7%) of crohn's disease, 19 cases (8.2%) of ulcer, 13 cases (5.6%) of intestinal vascular malformation, 12 cases (5.2%) of small intestinal stromal tumor. The lesion detection rate in obscure abdominal pain and obscure gastrointestinal bleeding were 59.6% (62/104) and 67.0% (63/94). In all patients, there were 1 case of small bowel perforation, the remaining patients had no serious complications such as bleeding and perforation. Conclusion The positive detection rate of double-balloon enteroscopy examination is high, and the double-balloon enteroscopy examination is relatively safe. So, double-balloon enterscopy examination has high diagnostic value for detecting small intestine diseases.

OBJECTIVETo compare the differentiation ability difference of hematopoietic, mesenchymal and endothelial potential between CD41⁺ cells derived from the mouse aorta-gonadmesonephros (AGM) region, yolk sac (YS) and embryonic circulating blood (CB).

METHODSCD41⁺ cells were sorted from AGM, YS and CB. The CD45 and c-kit expression were studied in CD41⁺ cells by flow cytometry. IL-3 and bone morphogenetic protein 4 (BMP-4) treatment together with semi solid culture were used to assess hematopoietic potential difference of CD41⁺ cells. Immunofluorescence staining of α-SMA was used to assess mesenchymal potential difference. The endothelial cell induction system was used to assess endothelial potential difference.

RESULTSThe proportions of CD45+ cells in CD41⁺ population were 51.9% (AGM), 45.8% (YS) and 22.2% (CB), respectively, while those of c-kit⁺ cells were 40.0% (AGM), 39.6% (YS) and 36.2% (CB), respectively. After stimulated by IL-3 factor, the number of total colonies increased in all three groups-derived CD41⁺ cells compared to that of unstimulated group[(14.1±1.9) vs (1.2±0.2), (32.4±1.1) vs (18.4±2.2) and (41.8±0.9) vs (10.4±1.8)], (P<0.01). After stimulated by BMP-4 factor, compared to unstimulated group, CFU-Mix colony number in CD41⁺ cells from AGM region and YS were significantly decreased[(0.5±0.6) vs (3.2±0.8), (1.3±0.7) vs (7.4±1.7)](P<0.01), but there was no difference in CB group[(2.5±0.5) vs (3.9±1.5)](P>0.01). The mesenchymal marker α-SMA was highly expressed in CD41⁺ cells from AGM region and YS, but lowly expressed in CD41⁺ cells from CB.

CONCLUSIONThere are some differences between CD41⁺ cells in AGM region, YS and CB on hematopoietic cell surface marker expression, hematopoietic colony formation with IL-3 and BMP-4 stimulation.

Animals ; Aorta ; cytology ; Bone Morphogenetic Protein 4 ; pharmacology ; Cell Differentiation ; Gonads ; cytology ; Interleukin-3 ; pharmacology ; Mesonephros ; cytology ; Mice ; Platelet Membrane Glycoprotein IIb ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Yolk Sac ; cytology