AIMTo simultaneously determine three unconjugated neurosteroids, dehydroepiandrosterone (DHEA) , pregnenolone (PREG), allopregnenolone (AP), from several brain regions of the rat.

METHODSNeurosteroids were isolated separately in a two steps procedure by using ethyl acetate-n-hexane (90:10) as the first step to extract the unconjugated steroids, then the steroid fractions were further purified by SPE. All steroids were derivatized with 2-nitro-4-trifluoromethylphenylhydrazine (2-NFPH) and analyzed by HPLC-MS ( APCI) using selected-ion monitoring. Methyltestosterone was chosen as the internal standard. Results The linear calibration curve of DHEA was obtained in the concentration range of 0.030-2.00 microg x L(-1). The linear calibration curves of PREG and AP were obtained in the concentration range of 0.025-2.00 microg x L(-1). The concentrations of DHEA, PREG and AP in male rat brain regions were (0.70 +/- 0.23), (4.8 +/- 1.9), (1.1 +/- 0.6) ng x g(-1) for frontal cortex, (0.57 +/- 0.28), (6 +/- 3), (0.5 +/- 0.3) ng x g(-1) for hippocampus, (1.5 +/- 1.0), (9 +/- 5), (1.4 +/- 0.9) ng x g(-1) for amygdale, (0.52 +/- 0.14), (7.7 +/- 2.8), (0.5 +/- 0.6) ng x g(-1) for striatum, (2.9 +/- 1.6), (18 +/- 9), (1.6 +/- 1.3) ng x g(-1) for nucleus accumbens, (4.0 +/- 2.0), (27 +/- 12), (0.8 +/- 0.5) ng x g(-1) for pituitary gland, (1.7 +/- 1.2), ( 16 +/- 10), and (0. 8 +/- 0.7) ng x g(-1) for hypothalamus, respectively.

CONCLUSIONGood linearity and accuracy were observed for each steroid. The procedure was suitable for measuring concentrations of the unconjugated steroids in rat brain simultaneously.

Amygdala ; chemistry ; Animals ; Brain Chemistry ; Chromatography, High Pressure Liquid ; Corpus Striatum ; chemistry ; Dehydroepiandrosterone ; analysis ; Hippocampus ; chemistry ; Hypothalamus ; chemistry ; Male ; Mass Spectrometry ; methods ; Nucleus Accumbens ; chemistry ; Prefrontal Cortex ; chemistry ; Pregnenolone ; analogs & derivatives ; analysis ; Rats ; Rats, Sprague-Dawley ; Sensitivity and Specificity

Objective To explore the relationships of serum leptin(LP),tumor necrosis factor-?(TNF-?)and insulin resistance in simple obesity children.Methods Forty-seven children with simple obesity were divided into two groups: high insulin group(HIG) and normal insulin group(NIG) according to the levels of fasting state of serum insulin and glucose.The fasting state of serum insulin(FINS),LP,TNF-? and fasting blood glucose(FBG) were measured.The relationships of LP,TNF-? and insulin resistance index(HOMA-IR) were also analyzed.Results 1.The BMI,waist,hip,waist hip ratio(WHR),LP,TNF-?,HOMA-IR were obviously higher in the HIG than those in the NIG and NCG groups(Pa0.05).3.In the HIG group,the FINS was positively correlated to LP,TNF-? and HOMA-IR(r=0.560,0.413,0.864 P

Objective To optimize the short tandem repeats(STR)which link closely to survival motor neuron(SMN)and have redundant polymorphism information contents,and to use these STR in the prenatal diagnosis of spinal muscular atrophy(SMA).Methods Eleven STR loci(D5S435,D5F153, DSF151,D5S637,D5S1413,D5S125,D5S464,D5S1556,DSF149,D5S351,MAP1B-5')were amplified by PCR.Then the PCR products were detected by polyacrylamide gel electrophoresis(PAGE)and analyzed by silver staining.STR loci were evaluated and optimized by their PIC values.PCR-PAGE and gene scan were combined to make genetic link analysis for SMA families based on the optimized STR.Results Three STR loci(D5S435,DSF149 and D5S351)were selected with 8,19 and 18 polymorphic fragments detected respectively in 100 normal individuals.Their PIC values were 0.84,0.91 and 0.92 respectively.Four carriers and 2 normal individuals were detected from 6 SMA families with linkage analysis by using the 3 STR.Conclusion This genetic diagnosis system based on the 3 STR loci can provide rapid prenatal diagnosis for SMA families,can eliminate maternal blood contamination,and also can discriminate carriers from normal individuals in the fetuses,which makes the prenatal diagnosis system of SMA perfect.

OBJECTIVETo study human vestibular cerebral representations by combining right-sided ice-water stimulation at 0 degree C with blood oxygenation level dependent functional magnetic resonance imaging (BOLD-fMRI) and to evaluate the value of this method in the functional localization of human vestibular cortex.

METHODSTwenty right-handed volunteers (12 men and 8 women) received unilateral irrigation of the right external auditory meatu for 15 s with 15 ml of water at 0 degrees C during fMRI in complete darkness. The functional imaging of brain cortex was acquired with a 1.5-T MRI scanner (Signa Infinity Twin + Excite; General Electric Co., USA). The successive functional images from each subject were analyzed as a group with statistical parametric mapping software (SPM 99).

RESULTSUltimately, data obtained from 17 subjects were analyzed (3 subjects were eliminated from data because of head movement exceeding 2 mm). The group analysis showed bilateral (particularly left-sided) cortical activation, associated with caloric stimulus involving in temporoparietal junction extending into the posterior insula, supramarginal gyrus in the inferior parietal lobe, precuneus, supplementary motor area (SMA), the ventrolateral portion of the occipital lobe, cuneus and lingual gyrus, superior temporal gyrus and cingular cortex.

CONCLUSIONSIce-water stimulation at 0 degree C in fMRI reveals a widespread cortical network involved in vestibular signal processing in human. As the functional localization of vestibular cortex could be determined precisely, ice-water stimulation at 0 degree C in fMRI would hold great promise as a sensitive and reproducible tool for the research in human vestibular cortex.

Adult ; Brain Mapping ; Cerebral Cortex ; anatomy & histology ; physiology ; Ear, Inner ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Vestibular Nuclei ; anatomy & histology ; physiology ; Young Adult

OBJECTIVETo investigate the effects of prostaglandin E1 (PGE1) on the progression of aristolochic acid nephropathy (AAN).

METHODSTwenty-four patients diagnosed as AAN with serum creatinine (Scr) between 1.5 mg/dL and 4 mg/dL during September 2001 to August 2003 were randomly divided into 2 groups. All patients had ingested long dan xie gan wan containing aristolochic acid (0.219 mg/g) for at least 3 months. Twelve patients were injected with Alprostadil (10 microg/d for 10 days in one month, summing up to 6 months). Except for PGE1, the other therapy was same in both groups. Renal function was assessed using reciprocal serum creatinine levels (1/Scr).

RESULTSThe level of Scr an d serum hemoglobin (Hgb) was similar in both groups prior to therapy. During follow-up, 1/Scr levels in PGE1 group were significantly higher than control group (P < 0.01), and Hgb levels in PGE1 group were significantly increased compared with control (P < 0.05).

CONCLUSIONPGE1 can slow the progression of renal failure and increase Hgb level of AAN patient.

Adult ; Alprostadil ; therapeutic use ; Aristolochic Acids ; adverse effects ; Creatinine ; blood ; Female ; Follow-Up Studies ; Hemoglobins ; metabolism ; Humans ; Kidney ; pathology ; Male ; Middle Aged ; Nephritis, Interstitial ; chemically induced ; drug therapy ; pathology

OBJECTIVETo introduce the application of denaturing high-performance liquid chromatography (DHPLC) in the diagnosis of childhood type spinal muscular atrophy (SMA).

METHODSExon 7 and flanking area of survival motor neuron (SMN) gene were amplified by PCR in 1 standard sample, 25 normal individuals and 25 patients with SMA. The PCR products were then directly loaded onto the DHPLC system after denaturing and annealing. Different DNA segments were separated by changing the concentration of buffer A relative to that of buffer B.

RESULTSDifferent DNA segments were separable on the DHPLC chromatogram. Three peaks including SMN1/SMN2 heteroduplex peak, SMN2 homoduplex peak and SMN1 homoduplex peak were detected in 23 out of 25 normal individuals. Only SMN1 homoduplex peak was detected in 2 normal individuals and the standard sample, indicating the deletion of SMN2 On the contrary, only the SMN2 homoduplex peak was detected in 22 out of 25 patients with SMA, indicating deletion of SMN1. The three peaks as those of normal individuals were detected in the other 3 patients, indicating no SMN1 or SMN2 deletion.

CONCLUSIONAs a new technology for diagnosing SMA, DHPLC is sensitive, accurate, rapid and convenient.

Chromatography, High Pressure Liquid ; methods ; Exons ; genetics ; Humans ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; SMN Complex Proteins ; genetics ; Sensitivity and Specificity ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein

OBJECTIVETo observe the effect of parecoxib on morphine dosage in patient-controlled analgesia (PCA) following thoracoscope-assisted thoracotomy.

METHODSA consecutive series of 100 patients undergoing thoracoscope-assisted thoracotomy were randomized into 5 groups and received PCA with morphine doses at 0, 5, 10, 15, and 20 mg given in 200 ml saline (groups P(1), P(2), P(3), P(4), and P(5), respectively). Parecoxib (40 mg) was given in all the patients immediately before the operation, and the mixture (4-5 ml) of lidocaine and ropivacaine was administered into the 3 intercostal spaces upper and lower to the incision before chest closure. PCA was administered for each patient. The visual analogue scale (VAS) at rest and coughing and the respiratory functional parameters were recorded at 1, 2, 4, 8, 12, 24, 36, and 48 h after the start of PCA, and the actual and effective button-pressing times (D(1)/D(2)) in PCA were also recorded.

RESULTSNo patients showed signs of respiratory inhibition within 24 h after the operation, and the resting VAS was comparable between the groups within the initial 6 postoperative hours. At 8 to 24 h postoperatively, the VAS scores at rest and coughing were significantly higher in P(1) group than in the other groups (P<0.05), and no significant differences were found between the groups at 36 to 48 h. D(1)/D(2) in groups P(1) and P(2) were significantly different from those in the other 3 groups at 4-24 h, but no such difference was found between groups P(3), P(4), and P(5).

CONCLUSIONThe application of parecoxib may reduce the dosage of morphine in PCA following thoracoscope-assisted thoracotomy and results in good analgesic effect without affecting the patients respiratory function and sputum elimination.

Adult ; Aged ; Analgesia, Patient-Controlled ; methods ; Combined Modality Therapy ; Double-Blind Method ; Female ; Humans ; Isoxazoles ; administration & dosage ; Male ; Middle Aged ; Morphine ; administration & dosage ; Pain, Postoperative ; drug therapy ; Thoracoscopy ; Thoracotomy ; methods ; Young Adult

OBJECTIVETo retrospectively compare the clinical outcome in patients with portal hypertension treated with transjugular intrahepatic portosystemic shunt (TIPS) using Fluency stent-graft (PTFE-covered stents) or bare stents.

METHODSApproval of study and treatment protocol and waiver of informed consent for the retrospective study were obtained from institutional review board. Informed consent was obtained from each patient before procedure. Sixty consecutive patients with portal hypertension treated with TIPS from April 2007 to April 2009 were included. TIPS creation was performed with Fluency stent-graft in 30 patients (group A) and with bare stents in 30 patients (group B). Liver function, TIPS patency and clinical outcome were evaluated every 3 months.

RESULTSDuring hospitalization, there was no hepatic encephalopathy and recurrency of variceal bleeding.Acute shunt occlusion was observed in one patient with group A and another patient with group B.Follow-up was performed with average time of (6.2 +/- 3.9) months in group A and (8.3 +/- 4.4) months in group B. The rates of recurrent bleeding, acute shunt occlusion, hepatic encephalopathy and death were 3.3% and 20.0%, 0 and 30.0%, 16.7% and 20.0%, 0 and 13.3% in group A and B. The rates of recurrent bleeding, acute shunt occlusion and death in group A was lower than those in group B. There was no difference of hepatic encephalopathy between group A and B. The decrease of portal pressure and portosystemic pressure gradient, and the increase of portal flow and shunt flow in group A were higher than those in group B. There were no difference of liver function, ammonia and MELD between group A and B.

CONCLUSIONSFluency stent-graft is safe and effective in TIPS creation, with high patency rate. Covered-stent can improve the clinical outcome of portal hypertension.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Hypertension, Portal ; surgery ; Male ; Middle Aged ; Portasystemic Shunt, Transjugular Intrahepatic ; instrumentation ; Retrospective Studies ; Stents ; Treatment Outcome ; Young Adult

BACKGROUNDEndothelial dysfunction is a key event in the onset and progression of atherosclerosis in diabetic patients. Apoptosis may lead to endothelial dysfunction and contribute to vascular complications. However, no study has addressed apoptosis in human umbilical vein endothelial cells (HUVECs) induced by an intermittent high-glucose media and its association with adiponectin receptor 1 (adipoR1), adipoR2, or adenosine monophosphate (AMP)-activated protein kinase (AMPK).

METHODSHUVECs were cultured in continuous normal glucose (5.5 mmol/L), continuous high glucose (25 mmol/L), alternating normal and high glucose and mannitol. In the alternating normal and high-glucose media, HUVECs were treated under different conditions. First, cells were transfected with the adipoR1-specific small-interfering RNA (siRNA) and then stimulated with globular adiponectin (gAD). Second, cells were cultured in both gAD and the AMPK activator 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR). Third, cells were cultured in the AMPK inhibitor adenine-9-β-D-arabino-furanoside (araA), gAD, and in AICAR.

RESULTSHUVEC apoptosis increased more significantly in an intermittent high-glucose medium than in a constant high-glucose medium. HUVEC apoptosis induced by an intermittent high-glucose medium was inhibited when the cells were pretreated with 3 µg/ml gAD, which rapidly activated AMPK and adipoR1 in HUVECs. However, adipoR2 was not activated.

CONCLUSIONSWe found that adipoR1, not adipoR2, is involved in mediating intermittent high-concentration glucose-evoked apoptosis in endothelial cells. gAD activated AMPK through adipoR1, leads to the partial inhibition of HUVEC apoptosis. A fluctuating glucose medium is more harmful than a constant high-glucose medium to endothelial cells.

AMP-Activated Protein Kinases ; antagonists & inhibitors ; genetics ; metabolism ; Adiponectin ; pharmacology ; Aminoimidazole Carboxamide ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Glucose ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; RNA, Small Interfering ; Receptors, Adiponectin ; genetics ; metabolism ; Ribonucleotides ; pharmacology ; Signal Transduction ; drug effects ; genetics

OBJECTIVETo explore the clinical value of "triple-rule-out" protocols using dual-source computed tomography for aortic dissection (AD) assessment.

METHODSTotally 25 patients suspecting of suffering from AD were examined on a dual-source computed tomography scanner. Two-dimensional and three-dimensional reconstruction was performed in all patients by means of multiplanar reconstruction, curved planar reformation, maximum intensity projection, and volume rendering. All images were read by two experienced radiologists in consensus. All patients were divided into AD group (n=12) and NO AD group (n=13) , The average Hounsfield unit values of true and false lumen were compared between superior of the aortic around the first endoleak and inferior of the aortic around renal artery.

RESULTSIn AD group, there were 6 patients with DeBakey type 1, 2 patients with DeBakey type 2, and 4 patients with DeBakey type 3. The image quality was rated on a 3-point scale as excellent in 10 patients (83.3%) and good in 2 patients (16.7%) . All cases was fully evaluable in NO AD group. The average Hounsfield unit values of true lumen between superior of the aortic around the first endoleak and inferior of the aortic around renal artery showed no significant difference between AD and NO AD group.

CONCLUSIONDual-source computed tomography offers a non-invasive, accurate, and rapid way to evaluate AD.

Adult ; Aged ; Aged, 80 and over ; Aneurysm, Dissecting ; diagnostic imaging ; Angiography ; methods ; Aortic Aneurysm ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult