Objective To develop a HPLC quantitatively test method of glycyrrhizin, liquiritin and isoliquritigenin simultaneously for licorice. Methods The 67% methanol extracts of licorice powder were analyzed by reverse-phase HPLC on an ODS column, using a gradient elution with a mixture of 1% H3PO4 and acetonitrile. The eluate was monitored by UV absorption at 248, 276, 360 and 370 nm. Results The linear equation of glycyrrhizin was Y =1?106X +16 220, r 2=1. The linear equation of liquiritin was Y= 2?106X+49 444, r 2=0.999 5. The linear equation of isoliquritigenin was Y =1?107X +4.466 7, r 2=1. The recovery rate of glycyrrhizin, liquiritin and isoliquritigenin was 98.01%, 102.63% and 98.18% respectively. Conclusion The method is accurate, stable amd reliable, and can be used to determine the three main chemical contents simultaneously.

Objective To retrospectively analyse the adverse reactions and complications of Graves' disease after thyroid arteries embolization.Methods 41 patients of Graves' disease underwent interventional embolization have been analysed with its adverse reactions and complications. Polyvinyl alcohol or bletilla microspheres and micro coils were used in these patients. Results Laryngopharyngeal and neck pain occurred in all patients. T 3 and T 4 increased in 3 days to one week after the procedure. Thirty of them showed fever. Dystopic embolism happened in two cases with one of transitory hypoparathyroidism. No hypothyroidism or hypoparathyroidism or hoarseness occur during long term follow up.Conclusions The adverse reactions and complications of Graves disease after thyroid arteries embolization may occur. Some of them are preventable and curable.

Objective To study the pathological morphology of thyroid specimen after thyroid arteries embolization. In order to select appropriate size of embolizing granules, the diameters of thyroid arteries in Graves' disease were measured. Methods Multiple slides of embolized thyroid tissues from superior pole, body and inferior pole of the resected thyroid glands were made. After being embedded and stained, pathological morphology was observed and diameters of arteries in various parts of thyroid glands were measured under microscopy. Results Pathological examination of thyroid glands showed that superior and inferior thyroid arteries and most of their branches were embolized with ischemic necrosis and fibrosis in the embolized thyroid tissue. Follicular epithelium appeared as flat or cubic shapes with colloid reduction. Average diameter of main branches of superior artery was 440～550?m and that of inferior artery was 300～375?m. The diameters of capillary network in the thyroid body was 120～250?m, and the non embolized ones was 40～110?m. The diameter of isthmus was 130～150?m. Conclusions Pathological morphology of Graves' disease after thyroid arteries embolization showed that the excretion of thyroid glands would be decreased and equivalents to subtotal thyroidectomy.

BACKGROUND: By now, safety of collagenase application is still controvertible and some scholars believed that collagenase might induce the peripheral tissue injury. It attracts much attention in clinic that whether there is nerve injury induced by collagenase chemonucleolysis (CCN) around the injection sites.OBJECTIVE: To observe the effects of collagenase on spinal nerve conduction velocity (NCV) of rats detected with evoked potential method so as to probe into the safetyof collagenase application and further demonstrate the safety of percutaneous intervertebral disc CCN. DESIGN: Randomly grouping design, animal experiment.SETTING: Department of Interventional Radiology, Affiliated First Hospital of Sun Yat-sen University. MATERIALS: The experiment was conducted in the Basic Medical College of Sun Yat-son University from July to September 2002. A total of 57 male SD rats were randomly divided into normal group (n=9), acute sham-operation group (n=10), subacute sham-operation group (n=8), chronic sham-operation group (n=7), acute experimental model group (n=9), subacute experimental model group (n=7) and chronic experimental model group (n=7). METHODS: After being anaesthetized by intraperitoneal injection of continal (45 mg/kg) to separate and identify dorsal root ganglion (DRG),rats in the experimental group were locally dripped with 1 mL of collagenase (300 U/ML) and those in the sham operation group locally dripped with 1 mL of normal saline. Stimulating electrode was placed in the A point of sciatic nerve and recording electrode in the B point of ganglionic central process segment of DRG in L5 nerve root. Evoked potentials A and B were simulated to continuously record latency twice, and the average value was calculated; Distance between A and B were measured and recorded. NCV = distance between A and B / latency. Evoked potential of a segment ofnerve including DRG was measured in the acute group at one hour after administration, in the sub acute group at one week after administration and in the chronic group at one month after administration. MAIN OUTCOME MEASURES: NCV of each group. RESULTS: A total of 57 enrolled animals were involved in the analysis. NCV in normal group, acute experimental group, acute sham-operation group, subacute experimental group, subacute sham-operation group, chronic experimental group, chronic sham-operation group were (45.4±10.7), (43.4±5.9), (46.3±6.5), (52.4±10.4), (49.7±8.1), (46.7±11.0) and (44.6±6.5) m/s respectively. There were no remarkable differences in NCV among all the groups by using one-way analysis of variance (F=1.010,P=0.430); It was showed that there were no marked differences in NCV between each two groups by using multiple comparisons (P=0.336). CONCLUSION: Collagenase at the therapeutic concentration applied in clinical CCN has no remarkable effects on NCV of rat spinal nerve, and to a certain extent, percutaneous intervertebral disc CCN is relatively reliable.

Objective To explore the mid to long term therapeutic efficacy of Graves′ disease after interventional embolization. Methods Twenty five patients of Graves′ disease treated with interventional embolization were followed up for 24-57 months. T 3 and T 4 were monitored at pre operation, six months, 12 months, 2, 3, and 4 years after operation, respectively. Other references included pulse, thyroid size, and vessel′s murmur. Results Twenty two patients completely relieved from the hyperthyroidism during the follow up. Only one patient suffered from recurrence. Other two patients were still on maintaining dosage of antithyroid drug therapy. No hypothyroidism or hypoparathyroidism was found during this term. Conclusion Mid to long term follow up showed satisfactory efficacy of interventional therapy, offering another alternative for treatment of refractory Graves′ disease.

Objective To develop an uncertainty study model of the enzymological reference method to systematically research the uncertainty influencing factors of 6 enzymological reference methods ALT,AST,LDH,AMY,GGT and CK for determining the key factors influencing the reference method and better guiding the establishment and operation of the enzymological reference methods.Methods The uncertainty of the six enzymological reference methods was evaluated by the theoretical evaluation com-bined with the experiment design according to series of standards including JJF1059-1999 Evaluation and Expression of Uncertainty in Measurement,JJF1135-2005 Evaluation of Uncertainty in Chemical Analytical Measurement,CANA-GL06 Guidance on Evalua-ting the Uncertainty in Chemical Analysis and CNAS-CL33 Guidance on the Application of Testing and Calibration Laboratories Competence Accreditation Criteria in the Field of Clinical Enzymology Reference Measurement.Results The established enzymo-logical uncertainty study model was C=ΔAΔt · 1ε · 1L ·V 1 +V 2 +V SV ·fT ·f pH ·fλ·fkit ·fresult .Conclusion The uncertainty e-S valuation model is set up and successfully applied in the uncertainty evaluation of the six enzymological reference methods.

BACKGROUND:There are many animal models used for studying discogenic pain, but percutaneous puncture annulus is rarely reported. Minimal y invasive approach to establish a discogenic pain model in Spraque-Dawley rats could reduce the interference factors of surgical trauma.
OBJECTIVE:To establish a Spraque-Dawley rat model of discogenic pain by percutaneous puncture annulus, with easy operations, high stability and obtaining large-scale productions, and to confirm the model by the results of behavior, MRI and molecular biology.
METHODS:Eighty-eight male Sprague-Dawley rats, of specific pathogen free level, were randomly divided into three groups, model group (n=44), control group (n=10) and sham group (n=34). In the model group, the annulus was percutaneously punctured under X-ray guidance;while rats in the sham group were punctured at the paravertebral tissue, rather than the annulus.
RESULTS AND CONCLUSION:The 50%mechanical withdrawal threshold of both hind paws in model group were reduced compared with control group and sham group. In the model group, the L 5/6 intervertebral disc degeneration was apparently visible, and the degree of degeneration was aggravated along the time. In the model group, the expression of calcitonin gene-related peptide in dorsal root ganglion of rats began to increase at 3 days post-operation and reached the peak at 21 days post-operation, then remained at high levels until the 35th day post-operation. The expression of tumor necrosis factor-αin dorsal root ganglion of rats increased at 3 days post-operation and reached the peak at 14 days post-operation, then remained at a higher level until the 35th day post-operation. The experiment result verifies that the discogenic pain model of Spraque-Dawley rats induced by percutaneous puncture annulus has the advantages of good stability and less trauma. The model can be used to study discogenic pain.

OBJECTIVE:To study the inhibitory mechanism of timosaponin B-Ⅱ(TB-Ⅱ) on the proliferation and migration of human lung cancer A549 cells. METHODS:A549 cells were treated with TB-Ⅱ [0(blank control),1,10 and 100 μg/ml] for 48 h,and total RNA and total protein were extracted respectively. Real time fluorescence quantitative-PCR and Western blot were used to detect mRNA and protein levels of IL-18. IL-18 in A549 cells was silenced by transfection;the expression of IL-18 mRNA and protein were compared among untransfection group,negative control group and transfection group;and then human lung can-cer A549 cells with silenced gene were treated with 10 μg/ml TB-Ⅱ for 24,48 and 72 h. The activity of cell proliferation was de-tected with CCK-8,and the change of cell migration ability was observed by streak method. RESULTS:Compared with blank con-trol,the expression of IL-18 mRNA and protein in A549 cells all increased after treated with TB-Ⅱ(P<0.05 or P<0.01),and were positively correlated with concentration. Compared with untransfection group,the expression of IL-18 mRNA and protein de-creased in transfection group(P<0.01). Compared with untransfected cell treated with TB-Ⅱ,the viability and migration ability of A549 cells with transfection gene increased after treated with TB-Ⅱ for 72 h(P<0.01). CONCLUSIONS:TB-Ⅱ can inhibit the proliferation and migration of A549 cells by up-regulating IL-18 gene expression.

BACKGROUND:Primates are considered to be the most appropriate animal model of lumbar intervertebraldisc degeneration, but the disc degenerated characteristics of monkeys were rarely reported. OBJECTIVE:To verify the degenerated regularity and characteristics of lumbar intervertebral disks in rhesus monkeys with magnetic resonance T2 mapping and T1ρimaging technology. METHODS:The sagittal lumbar intervertebral disc magnetic resonance T2 weighted imaging,T2 weighted mapping imaging and T1ρweighted imaging of 63 adult rhesus monkeys were acquired on 1.5T magnetic resonance equipment. The T2-map value and T1ρvalue of lumbar intervertebral disc regions of interest were calculated on the post-processing workstation. RESULTS AND CONCLUSION:(1) This study obtained 425 better magnetic resonance images of lumbar intervertebral disks in adult rhesus monkeys. T2-map value and T1ρvalue of nucleus pulposus were most consistent by different persons, and the Kappa coefficient was more than 0.93. (2) The T2-map value and T1ρvalue of nucleus pulposus were both negatively correlated significantly with Pfirrmann grades (r=-0.842, P<0.01;r=-0.896, P<0.01). The T1ρvalue and T2-map value of nucleus pulposus were significantly statistical y different between Pfirrmann grades I-IV (P<0.001, P<0.001). The T1ρvalue of nucleus pulposus was negatively correlated significantly with Pfirrmann grade II-III (r=-0.517, P<0.01) and Pfirrmann grade IV-V (r=-0.499, P<0.01). The T2-map value of nucleus pulposus was also negatively correlated significantly with Pfirrmann grade II-III (r=-0.617, P<0.01) and Pfirrmann grade IV-V (r=-0.652, P<0.01). (3) The T2-map value of L1-2 and L2-3 segments nucleus pulposus were significantly lower than that in L6-7 and L7-S1 segments (P<0.05). (4) There were significant differences in age among the T1ρvalue and T2-map value of nucleus pulposus (r=-0.702, P<0.001, r=-0.730, P<0.001). (5) It is concluded that magnetic resonance T2 mapping and T1ρimaging technology can objectively and sensitively assess the degenerated process of nucleus pulposus in rhesus monkeys. The degeneration in upper lumbar segments (L1-2 and L2-3) was earlier and more severe than that in lower lumbar segments (L6-7 and L7-S1) in rhesus monkeys. Age is one of the most important factors in lumbar intervertebral disc degeneration of adult rhesus monkeys.

OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.