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Objective: In the process of the study of human cancer, liver disease, ophthalmic disease, bone disease and other diseases, found in the blood of hyaluronic acid (HA) content is higher than normal. HA is mainly distributed in the mesenchymal cells, the combination of these HA expressed in form of a complex with a covalent bond with the cartilage chain protein (CLP). Certain concentrations of guanidine hydrochloride the CLP and HA dissociation, nobody reported pig cartilage chain protein (p-CLP) in vivo distribution and metabolic processes. To this end, we have discussed the following: ① The isolation of p-CLP; ② p-CLP, with HA vitro binding characteristics; (3) lung transplantation in nude mice with HA tumor learning distribution; ④ p-CLP in nude mice lung transplant tissue distribution. Hope that the above study, p-CLP biology and experimental nuclear medicine imaging studies underlying data to provide experimental evidence for the early diagnosis of lung tumor imaging and other diseases. Methods: The animal model of lung adenocarcinoma tumor learning and HA distribution in A549 cells inoculated BALB / C-nunu tumor-bearing nude mice were reared 3,4,5,6 weeks immunohistochemistry SP method determination of HA distribution within the tumor in the lung transplantation nude mice of different ages. 2. P-CLP separation, purification and identification of affinity chromatography and separation from porcine costal cartilage purified p-CLP, through epichlorohydrin activated Sepharose 4B Preparation of HA-AH-Sepharose 4B affinity chromatography, p-CLP crude products homogenate, enzymatic hydrolysis, dialysis, concentrated, and passed through a column purification made of p-CLP; then the SDS-PAGE gel electrophoresis analysis of the molecular weight of the p-CLP; p-CLP amino acid analysis to determine the content of amino acids, p-CLP; competition in the external beam radiotherapy combined analysis of the determination of the p-CLP HA affinity characteristics. Iodine mark of 3.p-CLP and tracer analysis the the Na125I chloramine-T oxidation tag p-CLP, and explore the optimal reaction conditions, preparation of high specific activity of the radioactive probe; tracer distribution experiment through to lung transplantation nude tail vein injection of 125I-p-CLP, 5min, 20min, 30min, 45min, 1h, 1.5h, 3h, 5h, 7h period of time the mice were sacrificed, blood and vital organs measuring radioactivity in the p-CLP nude mice in vivo metabolism and distribution parameters. Results: HA immunostaining positive rate of 93%, immunohistochemistry 3-6 weeks positive tissue optical density values ??were 150.78 ± 10.25,160.13 ± 8.54,176.83 ± 13.46,185.95 ± 13.61, the control group was 37.43 ± 5.17. The t-test showed that each group was statistically significant (P lt; 0.05). The immunohistochemistry Law vaccination 3-6 weeks in nude mice with the time extension of the vaccination, HA expression gradually increased, the HA is mainly distributed in the tumor cells surrounding stromal tumor HA expression rich, normal tissue expression was weak. 2 affinity chromatography and extraction p-CLP, the UV scanning into a single peak, the to extract protein yield was 2.21%, the SDS-PAGE electrophoresis p-CLP two bands, respectively (44.36 ± 2.90) KD p-CLP HA affinity constant (71.58 ± 4.13) KD; p-CLP amino acid analysis glutamate levels in the highest (2.26mg/20mg) tyrosine content 0.91mg/20mg; competitive binding analysis of external beam radiation experiments show Ka (6.30 ± 1.73) x 109M-1 3.p-CLP, mark display when 125I activity for 0.925MBq and there are significant differences the 1.85MBq 3.7MBq 7.4MBq groups labeling rate (all P lt; 05) the 7.4MBq 11.1MBq, 14.8MBq group labeling rate differences (all P gt; 0.05); chloramine-T concentration 0.312mg/ml stamp rate 0.625mg/ml with other groups differences (all P lt; 0, 05), other markers showed no difference (all P gt; 0.05); 30s-8min mark time mark rate in each group there was no difference (both P gt; 0.05); 125I activity concentration of chloramine-T, marking time on the radiochemical purity of little effect, each group no significant differences (all P gt; 0.05); chloramine-T concentration 0.625mg/ml and mark time 1min 125I-p-CLP biological binding highest activity; distribution of the p-CLP tracer experiments show that, when the concentration of the p-CLP in vivo 20min peak, with the time, p-CLP distribution within the tumor percentage gradually increased. Conclusion: 1.HA in lung tumor tissue, the increasing trend, wherein the periphery of the tumor cells, interstitial excessive expression, with the extension of the time of tumor growth, HA expression increased with the growth of the tumor cells. P-CLP labeling conditions: the 125I 7.4MBq 1.25mg/ml chloramine-T 30 μl p-CLP 0.5mg reaction volume of 100 μl of reaction 1min. Tracer experiments, p-CLP in early nude mice was systemic distribution, metabolism by the liver, in part by renal excretion, HA-specific binding in the nude mouse lung tumor model p-CLP and lung tumor cells in the interstitial .
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