A GC method is described for measuring fentanyl in man plasma. Fentanyl was extracted at basic pH with cyclohexane-isobutyl alcohol (197:3) with Ro21-2212 added as internal standard and fentanyl derivative as earner. The drug was back-extracted in H2SO4, then the extract was made basic and recxtracted with ethyl ether-dichloromethane (9:1). The organic phase was evaporated at 40℃ under N2 and the residue was dissolved in ethanol. A. portion (2?l) was analyzed on a wide-bore capillary column (10 m ? 0.53 mm) of HP-1 (2.65 ?m), operated at 255℃ with N2 as carrier gas (8 ml/min) and N -P detector. The calibration graph was linear within the ranges of 0.25-100 ng/ml with a corelation coefficient of 0.9996 and the detected limit was 0.2 ng/ml. Mean recovery was 99.02% ? 6.81%. The coefficient of variation for the within- and between-run were all less than 8%. No interference was found from endogenous compounds, metabolites of parent drug or other commonly used drugs. The method was applied to therapeutic drug monitoring and pharmacokinetic studies.

Objective To explore the immunophenotype features and folate receptor expression of blasts in patients with myelodysplastic syndromes(MDS).Methods Four-color flow cytometry using conventional and secondary gating strategies was used into analysis the immunophenotype features and folate receptor(FR) expression of blasts and CD+34 cells in bone marrow nucleated cells with MDS.The patients with acute myeloid leukemia-M2(AML-M2) were as positive control.Results with progression of MDS from RA/RAS,RAEB to RAEB-T,using conventional gating strategy,the proportion of CD+34 cells were gradually increased(P＜0.05).Moreover,the expression of HLA-DR,CDll7,CD13,CD33 were also gradually increased and the expression of CDl5 was gradually decreased(P＜0.05).Using secondary gating strategy,the expression of HLA-DR,CD117,CD13,CD33 on blasts were higher than those by conventional gating(P＜0.05).However,there was no significant difference(P＞0.05) in the expression of above mentioned antigens on CD+34 cells among different MDS subtypes.On the other hand,there were no expression of FR on blasts and cD+34 cells with different MDS subtypes.Conclusion With progression of MDS,the antigens of blasts surface change into more immature immunophenotype of medullary system.But these antigens abnormal expression only illustrates the increase of ascendant malignant clone quantity,it can not reflect the nature of the disease.Using flow cytometry technique can not detect whether or not FR expression on the blasts with MDS.

Objective To study the influence of Langchuang Ⅱ,a Chinese medicinal compound formula,on Th1/TH2cytokines of MRL/lpr lupus mice.Methods Forty MRL/lpr mice with spontaneous systemic lupus erythematosus(SLE) were randomized into blank control group,LangchuangⅡgroup(in the dose of 20 g?kg-1?d-1),prednisone group(in the dose of 6 mg?kg-1?d-1),and the combination group(LangchuangⅡand prednisone).After medication for 8 continuous weeks,Flowcytomix technology was used for the detection of different kinds of serum Th1/Th2 cytokines.Results Compared with the blank control group,serum granulocyte-macrophage colony-forming unit(GM-CSF) level was decreased in the combination group(P0.05);IL-4,IL-5 and IL-10 were all decreaed in the medication groups,and the decrease of IL-10 was obvious in the combination group(P

AIM: To study the effect of proliferating cell nucler antigen antisense oligonucleotide on ex vivo expansion of cord blood CD34~+ hematopoietic stem/progenitor cells.METHODS: CD34~+ cells were purified from fresh cord blood by immunomagnetic beads.CD34~+ cells were incubated in liquid culture system with different concentrations of(PCNA-ASODN).Using flow cytometry,the number of different kinds of stem/progenitor cells and PCNA expression were measured after CD34~+ cell incubation.RESULTS: PCNA was lowly expressed in low experiential group,with a positive rate of(27.2?3.6)% and(19.0?1.5)%,the positive rate of control group was(53.8?8.3)%(P

Horseradish peroxidase (HRP) was injected unilaterally or bilaterally into the cerebellum of 11 rabbits in order to trace the distribution of labeled neurons in the whole length of the spinal cord. To investigate the ascending side of the axons, hemisections were made unilaterally in the lower thoracic cord of 4 rabbits before injection. The distribution of labeled spinocerebellar tract neurons was rather wide. Neurons in the cervical segments were located in (1) the central cervical nucleus (CCN) in Q_(1-4), (2) the medial part of lamina VI in C_2-T_1, (3) the central part of lamina VII in C_(4-8) and (4) lamina IV-V in C_(5-8). The labeled neurons located in segments caudal to thoracic cord could be divided into two groups. Neurons of the uncrossed tract were located in (1) Clarke's column in T_2-L_4, (2) laminae IV-VI in T_2-L_6. Neurons of the crossed tract were found in (1) the spinal border cells (SBC)in L_(3-6), (2) the medial part of the lamina VII in segments caudal to L_6, (3) lamina V in the sacrococcygeal cord and (4) laminae VII-VIII in the sacrococcygeal cord. The present study suggests that the location and fiber course of the spinocerebellar neurons in the rabbit are quite the same as those in the cat. These results should form a basis for further anatomical and physiological studies of spinocerebellar system in the rabbit.

We have designed and carried out problem based learning (PBL) pedagogy since 2004. According to clinical eases, students learnt the pathophysiology of heart failure knowledge by themselves. Each group recommen-ded one student to make an oral presentation and wrote a review about heart failure. Preparing clinical cases and group discussions are very important. At the same time we should pay attention to the change in role of the teacher in PBL and cooperation with other disciplines.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven eases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi- quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpeint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperurieacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial- tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average foUow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16～89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.

OBJECTIVE: To evaluate the diagnostic accuracy of a dual-source computed tomography (DSCT) coronary angiography, with a particular focus on the effect of heart rate and calcifications. MATERIALS AND METHODS:One hundred and nine patients with suspected coronary disease were divided into 2 groups according to a mean heart rate (< 70 bpm and > or = 70 bpm) and into 3 groups according to the mean Agatston calcium scores (< or = 100, 101-400, and > 400). Next, the effect of heart rate and calcification on the accuracy of coronary artery stenosis detection was analyzed by using an invasive coronary angiography as a reference standard. Coronary segments of less than 1.5 mm in diameter in an American Heart Association (AHA) 15-segment model were independently assessed. RESULTS: The mean heart rate during the scan was 71.8 bpm, whereas the mean Agatston score was 226.5. Of the 1,588 segments examined, 1,533 (97%) were assessable. A total of 17 patients had calcium scores above 400 Agatston U, whereas 50 had heart rates > or = 70 bpm. Overall the sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) for significant stenoses were: 95%, 91%, 65%, and 99% (by segment), respectively and 97%, 90%, 81%, and 91% (by artery), respectively (n = 475). Heart rate showed no significant impact on lesion detection; however, vessel calcification did show a significant impact on accuracy of assessment for coronary segments. The specificity, PPV and accuracy were 96%, 80%, and 96% (by segment), respectively for an Agatston score less than 100% and 99%, 96% and 98% (by artery). For an Agatston score of greater to or equal to 400 the specificity, PPV and accuracy were reduced to 79%, 55%, and 83% (by segment), respectively and to 79%, 69%, and 85% (by artery), respectively. CONCLUSION: The DSCT provides a high rate of accuracy for the detection of significant coronary artery disease, even in patients with high heart rates and evidence of coronary calcification. However, patients with severe coronary calcification (> 400 U) remain a challenge to diagnose.
Calcinosis/*radiography ; Coronary Angiography/methods/*standards ; Coronary Disease/*radiography ; Coronary Vessels/*pathology ; Female ; *Heart Rate ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Sensitivity and Specificity ; Tomography, X-Ray Computed/methods/*standards

Objective: To investigate the characteristics, diagnosis, primary detection, and prognosis of cervical lymph node metastases of squamous cell carcinoma of unknown primary site (SCCUP). Methods: This study retrospectively analyzed the clinical features and follow-up data of 262 patients with SCCUP. The Chi-square test were used to analyze the clinical performances, characteristics of pri-mary lesions, and sensitivity and specificity of examinations to identify original lesions. Factors related to the overall survival (OS) and progression-free survival (PFS) were also analyzed. Results: The 262 patients with SCCUP comprised more men, with a median age of 57 years. At the follow-up, 70 patients were diagnosed with primary lesions (26.7%), and the detection rates of primary lymph nodes in those who were male (30.1%), with a single lesion site (31%), and with levelⅣdisease (39.3%) were higher than those in patients who were female (17.4%), with multiple lesion sites (18.7%), and with levelⅡ/Ⅲdisease (20.8%). Compared with traditional imaging examinations, 18F-fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) had higher sensitivity and speci-ficity in detecting the primary tumor. Survival analysis showed that distant metastasis was an independent risk factor affecting OS and PFS, and the effect of N stage on PFS was statistically significant. Conclusions: In SCCUP patients, the proportion of patients who were male, with a single lesion site, and with cervicalⅣlymph node metastasis had higher rates of detection of the primary sites. PET/CT examination is important for the diagnosis of SCCUP, as well as the detection of primary lesions. Advanced N stage and distant metasta-sis indicated poor prognosis.