Objective To investigate arterial blood gas(ABG)changes induced by different intra- abdominal pressures during CO_2 pneumoperitoneum in rabbits with lung and liver impact injury under controlled hemorrhage.Methods In this study 75 rabbits were randomized into nine groups(6 rabbits in each group)according to the amount of blood loss(6 ml/kg、12 ml/kg、22 ml/kg)and intra-abdominal pressures(5 mm Hg,10 mm Hg,15 mm Hg).After model was established successsfully,respiratory rates (RR),ABG and death rates were observed at pre-pneumoperitoneum,after 0.5 hour and 2 hours under pneumoperitoneum and 0.5 hour after desufflation.Results Without pneumoperitoneum,different blood loss exerted different effect on arterial blood gas:RR,PaCO_2 increased(P

To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 μg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Contamination ; Mycotoxins ; analysis ; Panax notoginseng ; chemistry ; Tandem Mass Spectrometry

Objective To assess and compare the difference in image quality and exposure dose between single-sided reading image plate(IP)and dual-sided reading IP.Methods A contrast-detail phantom CDRAD 2.0 was exposed by single-sided and dual-sided reading IP with different mAs sets.The entrance surface doses were recorded for all images.Images were then presented to two radiologists on a high resolution monitor of diagnosis workstation.The image quality figure(IQF)was measured for each image. Statistical analysis was performed using Spearman's correlation test and Wilcoxon signed-rank test to compare the difference in image quality and exposure dose between single-sided IP and dual-sided reading IP. Results With different tube current dosage of 5.6,12.0,20.0,25.0,and 40.0 mAs,IQF values of single-sided reading IP were 47.95,37.68,34.31,28.61,and 24.65,respectively,while those of dual- sided reading IP were 38.83,29.81,29.65,25.16,and 21.43,respectively.The IQF difference between them showed statistical significance(P

OBJECTIVETo investigate whether there is endogenous neural stem cell proliferation and whether these proliferated neural stem cells represent neural plasticity in the adult rats after cerebral infarction.

METHODSCerebral infarction models of rats were established and the dynamic expression of bromodeoxyuridine (BrdU), BrdU/polysialylated neural cell adhesion molecule (PSA-NCAM) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark dividing neural stem cells. PSA-NCAM was used to mark the plasticity of neural stem cells.

RESULTSCompared with controls, the number of BrdU-positive cells in the subventricular zone (SVZ) and hippocampus increased significantly at 1st day after cerebral infarction (P < 0.05), reached maximum at 7th day, decreased markedly at 14th day, but it was still elevated compared with that of the controls (P < 0.05). The number of BrdU-labeled with PSA-NCAM-positive cells increased significantly at 7th day (P < 0.05), reached maximum at 14th day, markedly decreased at 28th day, but it was still elevated compared with that of the controls (P < 0.05). It was equal to 60% of the number of BrdU-positive cells in the same period.

CONCLUSIONCerebral infarction may stimulate the proliferation of endogenous neural stem cells in situ and most proliferated neural stem cells represent neural plasticity.

Animals ; Bromodeoxyuridine ; metabolism ; Cell Proliferation ; Cerebral Infarction ; metabolism ; pathology ; Cerebral Ventricles ; pathology ; Hippocampus ; pathology ; Male ; Neural Cell Adhesion Molecule L1 ; metabolism ; Neuronal Plasticity ; Neurons ; metabolism ; pathology ; Rats ; Rats, Wistar ; Sialic Acids ; metabolism ; Stem Cells ; metabolism ; pathology

OBJECTIVETo study the effect of ZGDHu-1 on proliferation, differentiation and apoptosis in SHI-1 human leukemia cell line and explore its possible mechanism. Methods SHI-1 cells were cultured with different concentration of ZGDHu-1 and for different time. The cell proliferation was analysed by cell counting, alive cell count, MTT assay and Brdu-ELISA. Cell apoptosis was analysed by morphology, DNA content, Annexin-V/PI and Hoechst 33258 labeling method. Cell differentiation were assayed by morphology,expression of CD11b,CD14 and CD64 and NBT reduction. The expressions of phosphorylated p38MAPK or STAT3 were analysed by flow cytometry.

RESULTSZGDHu-1 inhibited SHI-1 cell proliferation in a time and dose dependent manner, the IC50- 48 h and IC50- 72 h were 250 ng/ml and 85 ng/ml, respectively. The majority of SHI-1 cells were arrested in G2/M phase. 48h after treated with 200 ng/ml ZGDHu-1, and those in G2/M phase accounted for (48.4 +/- 2.1)%. The SHI-1 cells apoptosis was increased with a time- and does-dependent manner. The morphology of SHI-1 cells cultured with 2-50 ng/ml ZGDHu-1 for three days become more mature with higher NBT positivity and up-regulated expressions of CD11b,CD14 and CD64. The expression of phosphor-p38MAPK was increased and phosphor-STAT3 down-regulated by the treatment of ZGDHu-1.

CONCLUSIONZGDHu-1 can inhibit SHI-1 cell proliferation and induce the cell differentiation and apoptosis. The mechanism may associate with its up-regulation of phosphor-p38MAPK and down-regulation phosphor-STAT3.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Formamides ; pharmacology ; Heterocyclic Compounds, 1-Ring ; pharmacology ; Humans ; Leukemia ; pathology ; Phosphorylation ; STAT3 Transcription Factor ; biosynthesis ; p38 Mitogen-Activated Protein Kinases ; biosynthesis

This study was aimed to investigate the changes of reactive oxygen species and antioxidative capacity on nitric oxide induced apoptosis in HL-60 cells. By means of in vitro incubation of HL-60 cells with sodium nitroprusside (SNP), the growth inhibition was detected by MTT assay. Cell morphology was observed by transmission electronmicroscopy and light microscopy. The apoptosis was analyzed by DNA agarose gel electrophoresis, DNA content and Annexin-V/PI labeling method. Reactive oxygen species (ROS) labeled with dihydrorhodamin 123 in cells was determinated by flow cytometry. The SNP-treated cells were examined for glutathione (GSH) level and activity of catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidase (GPX). The results indicated that SNP could inhibit HL-60 cell growth. Cell apoposis was confirmed by typical cell morphology, DNA fragment, sub-G(1) phase and Annexin-V/PI labeling method. HL-60 cell apoptosis was induced by SNP in a dosage- and time-dependent manner. After exposing to SNP at the concentration of 0.5 - 3.0 mmol/L for 48 hours, the mean fluorescence intensity of ROS in cells was significantly higher than those in groups control and potassium ferricyanide (PFC). During the apoptosis process, level of ROS in cells increased, levels of GSH, CAT, GPTand GPX decreased. The significant dose-effect relationship existed between the levels of ROS, CAT, GST, GPX and SNP dose. It is concluded that change of intracellular reactive oxygen species and antioxidative capacity are an important factors during the process of SNP-induced apoptosis in HL-60 cell.
Antioxidants ; metabolism ; Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; HL-60 Cells ; Humans ; Microscopy, Electron ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Reactive Oxygen Species ; metabolism

OBJECTIVETo establish a comprehensive quality control method for total flavonoid of Fructus Aurantii.

METHODSRP-HPLC and spectrophotometry were applied for the quantitative and fingerprint analysis of total flavonoid of Fructus Aurantii. The contents of naringin and neohesperidin were determined on an Agilent SB-C₁₈column (4.6 mm × 250 mm, 5 μm). The mobile phase was composed of 0.02 % H₃PO₄ and CH₃CN (80:20). The flow rate was 1 ml/min with DAD detected at 280 nm. The column temperature was maintained at 35°C. The fingerprints were developed on an Agilent SB-C₁₈ column (4.6 mm × 250 mm, 5 μm). The mobile phase was composed of 0.5 % HAc and CH₃OH with a linear gradient elution. The ratio of 0.5 % HAc and CH₃OH was: 0 min, 80:20; 10 min, 60:40; 35 min, 30:70; 50 min, 0:100. The flow rate was 1 ml/min with DAD detected at 320 nm. The column temperature was maintained at 30 degree. Meanwhile, the contents of total flavonoid were determined at 283 nm.

RESULTThe contents range of naringin, neohesperidin and total flavonoid were 38.3 %- 47.2%, 21.0 %- 28.5% and 79.9%-88.6 %, respectively. The fingerprints of the effective fractions showed 12 common peaks and the fingerprint similarity was all above 98.0 % compared with the standard chromatogram.

CONCLUSIONThe method reported in this paper can be used effectively for the quality control of total flavonoid of Fructus Aurantii.

Chromatography, High Pressure Liquid ; methods ; Citrus ; chemistry ; Flavonoids ; analysis ; Quality Control

To study the molecular mechanisms of nitric oxide donor sodium nitroprusside (SNP) -induced apoptosis in K562 human leukemia cell line, the different concentrations of SNP and different time of culture were used to treat K562 cell. At the same time, potassium ferricyamide (PFC) was used as control, blank was designed in experiment. Cell apoptosis was analysed by cell morphology, DNA agarose gel electrophoresis, DNA content, and annexin-V/PI labeling method. The TdT-mediated dUTP nick end labeling (TUNEL) assay was used to quantify in situ cell apoptosis. Reactive oxygen species (ROS) in cells and mitochondrial transmembrane potential (DeltaPsim) were labeled by dihydrorhodamin 123, 2', 7'-dichlorodihydrofluorescein diacetate and rhodamin 123/PI. bcl-2, bax, bad, p53 gene proteins and mitochondrial membrane protein were analysed by flow cytometry. The results showed that the K562 cell apoptosis was confirmed by typical cell morphology, DNA fragment, sub-G(1) phase, TUNEL and annexin-V/PI labeling. A majority of K562 cells were arrested in G(0)/G(1) phase. During the process of SNP-induced apoptosis in K562 cell, the mean fluorescence intensity of ROS in cells was significantly higher than those in blank and PFC control, while the DeltaPsim reduced. The expression of p53, bax, bad, Fas protein and mitochondrial membrane protein increased and bcl-2 protein decreased after SNP treatment. It is concluded that SNP induces K562 cell apoptosis through increasing ROS in cells, expressing the p53, bax, bad, Fas protein and mitochondrial membrane protein and decreasing bcl-2 protein, opening the mitochondrial permeability transition pore and reducing DeltaPsim. Furthermore, the Fas was activated during the apoptosis process.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Humans ; In Situ Nick-End Labeling ; K562 Cells ; Membrane Potential, Mitochondrial ; drug effects ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; biosynthesis

OBJECTIVETo investigate the changes of neural stem cells (NSCs) in the rat hippocampus after cerebral infarction (CI) and to evaluate the neurogenesis caused by the activation of NSCs.

METHODSCI models of rats were made and rats were assigned to 6 groups: sham-operated, 1 day, 3 days, 7 days, 14 days, and 28 days after CI. The dynamic expression of bromodeoxyuridine (BrdU), polysialylated neural cell adhesion molecule (PSA-NCAM), glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark the proliferated NSCs. PSA-NCAM was used to mark the plasticity of activated NSCs. GFAP and NeuN were used to mark the differentiated NSCs.

RESULTSCompared with the controls, the number of BrdU+ cells in the hippocampus increased significantly at 1 day after CI (P < 0.05), reached peak at 7 days after CI (P < 0.05), decreased but still elevated compared with the controls at 14 days after CI (P < 0.05), and nearly unchanged at 28 days after CI. The number of BrdU+/PSA-NCAM+ cells increased significantly at 7 days after CI (P < 0.05), reached peak at 14 days after CI (P < 0.05), and decreased but still elevated compared with the controls at 28 days after CI (P < 0.05). The number of BrdU+/PSA-NCAM+ cells was equal to 60% of the number of BrdU+ cells in all the same period. The number of BrdU+/NeuN+ cells in the hippocampus increased significantly at 14 days after CI (P < 0.05) and reached peak at 28 day after CI (P < 0.05). The number of BrdU+/GFAP+ cells in the hippocampus nearly unchanged after CI.

CONCLUSIONCI can stimulate the proliferation of inherent NSCs, and most proliferated NSCs may differentiate into neurons and represent neural plasticity.

Adult Stem Cells ; cytology ; metabolism ; Animals ; Bromodeoxyuridine ; metabolism ; Cell Nucleus ; pathology ; Cerebral Infarction ; metabolism ; pathology ; Dentate Gyrus ; cytology ; metabolism ; Glial Fibrillary Acidic Protein ; metabolism ; Hippocampus ; cytology ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Neural Cell Adhesion Molecule L1 ; metabolism ; Neurogenesis ; physiology ; Neurons ; cytology ; metabolism ; Rats ; Rats, Wistar ; Sialic Acids ; metabolism

OBJECTIVETo investigate the effects of CO(2) pneumoperitoneum on blood flow volume of abdominal organs of rabbits with controlled hemorrhagic shock model and liver impact injuries.

METHODSAfter controlled hemorrhagic shock and liver impact injuries, the rabbit model was established. Eighteen rabbits subjected to hemorrhagic shock and liver impact injuries were divided into 3 groups randomly according to the volume of lost blood: light hemorrhagic shock (blood loss volume was 10%, 6 ml/kg), moderate hemorrhagic shock (20%, 12 ml/kg) and severe hemorrhagic shock (40%, 22 ml/kg). Intraabdominal pressures of CO(2) pneumoperitoneum was 10 mmHg. Color-labeled microspheres were used to measure the blood flow volume of the liver, kidney and stomach before pneumoperitoneum at 30 minutes and 2 hours after pneumoperitoneum and 30 minutes after deflation. And the mortality and hepatic traumatic condition of rabbits were recorded.

RESULTSOf the 18 rabbits, there were 9 with liver impact injuries at Grade I , 8 at Grade II and 1 at Grade III (according to AIS-2005). The mortality rate in light hemorrhagic shock group was 33.33%, and that in moderate or severe hemorrhagic shock group was 100% within 30 minutes and 2 hours after pneumoperitoneum, respectively. The blood flow volume in the organs detected decreased at 30 minutes under pneumoperitoneum in light and moderate hemorrhagic shock groups. At the same time, the blood flow volume of the liver in moderate hemorrhagic shock group decreased more significantly than that in light hemorrhagic shock group.

CONCLUSIONSThe blood flow volume of abdominal organs in rabbits is decreased obviously under CO(2) pneumoperitoneum, with fairly high mortality rate. It is believed that CO(2) pneumoperitoneum should cautiously be used in abdominal injury accompanied with hemorrhagic shock, especially under non-resuscitation conditions.

Abdominal Injuries ; diagnosis ; Animals ; Blood Volume ; Carbon Dioxide ; Female ; Liver ; injuries ; Male ; Pneumoperitoneum, Artificial ; Rabbits ; Regional Blood Flow ; Shock, Hemorrhagic ; physiopathology