Angiogenesis Inhibitors ; pharmacology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Phytotherapy

BACKGROUND: Chitosan and sodium alginate are the good natural materials for microcapsule, and also used widely in tissue engineering. Our research teams have made thorough work at anticoagulant materials, but these materials are inert or simulate the liquid crYstal form of blood vessel wall. While in this experiment, on the base of our previous study, we microencapsulated heparin with biotic anticoagulation activity and other specific performances in order to enable microcapsule to have a long time releasing effect of medicine.OBJECTIVE: To microencapsulate the low molecular heparin so as to ensure the stability of heparin in vivo and analyze the effect of content of chitosan on the performance of heparin microcapsules basing on the natural chitosan and sodium alginate as the enwrapped materials of microcapsules.DESIGN: Open experiment.SETTING: Department of Material Science and Engineering, Jinan University.MATERIALS: The experiment was performed at the laboratory of Department of Material Science and Engineering, Jinan University from October 2004 to June 2005. Heparin, with relative molecular mass＜ 5 000, was provided by Shandong Freda Biochem Co., Ltd.,; Chitosan was provided by Shanghai Bio Life Science & Technology Co., Ltd, DD≥90%, η＜ 100 cps;Sodium alginate was provided by Qingdao Bright Moon Seaweed Industrial Co., Ltd. Emulsions were Span80, and CaCl2, which were both made in China.METHODS: ①Preparation of heparin/chitosan microcapsules (HCM):Some heparin aqueous solution was emulsified in liquid paraffin. The reaction system was stirred fully and presented emulsion. Then the whole reaction system was warmed to be at 50 ℃ and maintained for 20 minutes. Afterwards, 20 g/L chitosan solution was added slowly, subsequently with raising the temperature to be at 60 ℃ and then glutaraldehyde was dropwised keeping the reaction system at 80 ℃ for 1hour. Centrifugation, filtration and washing followed by washing with kerosene fully, remain organic was extracted by dehydrated alcohol with extractor were performed.Drying and xeransis in vacuum were done at last. ② Preparation of heparin-sodium alginate-chitosan microcapsules (HSCM) :Heparin aqueous solution and sodium alginate were emulsified in paraffin, and the reaction system was stirred into emulsion at room temperature for 20 minutes, then 3% CaCl2 solution containing different concentrations of chitosan was added slowly. 30 minutes later, Microcapsules were separated, washed and dried as the treatments as before. ③ Drug content and envelope efficiency were measured, heparin standard curve was determined and in vitro releasing effect of heparin microcapsules was also measured.MAIN OUTCOME MEASURES: ①Effect of chitosan solution concentration on preparation of heparin-chitosan microcapsules; ② Effect of glutaraldehyde dosage on preparation of heparin-chitosan microcapsules; ③Effect of sodium alginate concentration on hepatin-sodium alginate; ④Effect of chitosan concentration on hepatin-sodium alginate-chitosan microcapsules. ⑤ In vitro release of heparin microcapsules enwrapped by different materials. ⑥Measurement of heparin content and envelope efficiency. ⑦ Observation of heparin microcapsule under scanning electron microscope RESULTS: ①With the increasing concentration of chitosan, the color of production changed from yellow to dark, and microcapsules were increscent, but the microcapsules uniformity and property of balling were increased. ②The increasing content of glutaraldehyde led darker production.Increase of glutaraldehyde content made production bond each other severely. The glutaraldehyde, which did not react with chitosan, can solidify itself and presented anomalous microcapsules forming. ③There was not obvious balling property of the production with the change of concentration of sodium alginate. ④The balling property of microsphere was good with increasing concentration of chitosan. However, microcapsules conglutinated with each other. 2% chitosan would be better. ⑤With the increase of chitosan content, the releasing speed ofheparin became slow. ⑥The envelope efficiency was about 58% when microcapsule contained 20%(wt) of chitosan, and used chitosan only the envelope efficiency could approach to 79.9%. ⑦ The surface of microcapsules with chitosan was very compact,and with increasing of content of glutaraldehyde, microcapsules would bond each other.CONCLUSION: Chitosan at certain concentration will affect the uniformity and balling property of microcapsules. Chitosan dosage can alter the envelope efficiency of heparin. Envelope efficiency of heparin is increased and releasing speed of heparin is decreased with the increase of content of chitosan.

Objective: To investigate the dynamic expression of chemokines Mig, IP10, and ITAC after liver transplantation and to study its role in early diagnosis of acute rejection in humans. Methods: Thirty patients receiving liver transplantation (April 2005 to September 2005) were divided into acute rejection (AR) group (n=9) and non-acute rejection (NAR) group (n=18) based on the clinical symptoms and pathological examination. Three patients were excluded due to post-operation infection. The chemokines expression was determined in all patients 1 day before and day 1, 3, 5, 7 after transplantation. Sixteen patients with liver cirrhosis (cancer) and 16 normal adults were also examined as control in this study. Patients in AR group received pulse glucocorticoid treatment from the onset of AR and the expression of chemokines was determined on the day of AR diagnosis and day 3, 7 after glucocorticoid treatment. The relationship between Banff rejection activity index (RAI) and 3 chemokines expression on the day of AR onset was analyzed. Results: Chemokines expression was not significantly different between transplantation group and liver cirrhosis (cancer) group one day before transplantation; however, it was significantly higher than that of the normal control group (P<0.01). The expression of Mig, IP10 and ITAC was increased markedly in AR and NAR group day 3 after transplantation (P<0.05). AR was confirmed in 9 patients on day 11, 12 and 14 after transplantation. The serum contents of Mig, IP10 and ITAC in AR group were higher than those in NAR group at all defined time points after transplantation. There was a positive relationship between RAI and the expression of Mig, IP10 and ITAC on the day of AR onset (r=0.88, 0.94, 0.80, respectively). In AR patients, the expression of Mig, IP10 and ITAC decreased after pulse treatment with glucocorticoid (P<0.01). Conclusion: Serum level of Mig, IP10 and ITAC can serve as a sensitive, specific marker for early predication of AR in liver transplant patients.

Objective: To investigate the mechanism of CTIL-4Ig combined with Anti-ICAM-1 mAb in promoting immune tolerance induced by donor-derived immature dendritic cells (imDC) in recipient mice. Methods: Male mice were divided into 4 groups: control group (receiving only imDC), CTLA-4Ig group, ICAM-1 mAb group and CTLA-4Ig + ICAM-1 group. Mice were transfused with donor-derived imDC 7 days before they received heart transplantation in company with daily injection of ICAM-1 mAb, CTLA-4Ig or both for the following 2 weeks. Immunological analysis was performed in mice 7 days and 21 days after heart transplantation. Results: CTLA-4Ig alone or in combination with ICAM-1 mAb significantly inhibited T cells proliferation to alloantigen stimulation, impaired lymphocyte cytotoxicity, suppressed production of IL-2, IFN-γ by Th1, increased production of IL-10, and obviously decreased the production of alloantibody IgG in recipient mice treated with donor-derived imDC. ICAM-1 mAb alone had no significant effects on T cells proliferation and production of Th-derived cytokines except for IL-2. Conclusion: ICAM-1 mAb combined with CTLA-4Ig can enhance immune tolerance induced by donor-derived imDC in recipient mice through induction of T cells hypo-responsiveness, inhibition of lymphocyte cytotoxicity and B cell immunoreation, and promotion of Th2 polarization in vivo.

Objective:To study the expression and correlation of tumor-associated macrophages(TAM) and CCL5 in ganstric cancer.Methods:48 cases patients with completed clinical and pathological data of gastric cancer paraffin block specimens were select-ed.Cancer tissues and adjacent tissues were used as control,using SP immunohistochemical method to detect CD68 and CCL5 in gastric cancer tissues and adjacent tissues,and using the Spearman correlation statistics statistical methods for the correlation.Results:CD68 and CCL5 showed positive expression in gastric cancer tissue,significantly higher than those in the adjacent tissues(P<0.01),CD68 and CCL5 were related with gastric cancer invasion depth, lymph node metastasis, TNM stage and tumor differentiation ( P<0.001 ) . There was positive relation between the expression of CD68 and CCL5 in gastric cancer(P<0.01,r=0.759).Conclusion: CD68 and CCL5 played a driving role to the invasion and metastasis of gastric cancer occurrence,suggesting that the secretion CCL5 by TAM may promote the invasion and metastasis of gastric cancer.

AIM:To establish the fingerprint for Cacumen Platycladi (carbonized) by HPLC. METHODS:The column of Lichrospher C 18 (250 mm?4.6 mm 5 ?m)was used. The mobile phase consisted of 0.5‰ trifluoroactic acid-methanol with gradient elution. The detective wavelength was at 375 nm,and the flow rate was 1.0 mL/min. Different habitats were compared by Similarity Evaluation System for Chromatographic Fingerprint of CMM Version 2004A. RESULTS:The fingerprint consisted of 14 common peaks. The mutual mode of HPLC fingerprints was set up and the similarity of the crude drugs was in the range of 0.178-0.963. The standard HPLC fingerprint of Cacumen Platycladi (carbonized) was established too. CONCLUSION:This method is accurate and reliable and provides a scientific basis for the quality control of Cacumen Platycladi (carbonized).

Objective To investigate the diagnostic values of different imaging techniques in congenital esophageal atresia and tra-cheoesophageal fistula.Methods Imaging findinfs of 34 infants with congenital esophageal atresia and tracheoesophageal fistula were retrospectively analyzed.34 infants undertook X-ray and esophageal imaging,and 1 7 infants also undertook CT examination.Results Based on Gross classification,there were 2,32,9 and 23 cases for type I,Ⅲ,Ⅲa and Ⅲb,respectively.Coexistence of other de-formity was observed in 10 cases.Plain chest and abdomen X-ray film showed gastric pneumatosis in 32 cases,varied degrees of pneumonia in 25 cases and right upper pulmonary atelectasis in 3 cases.Esophagography showed a cecum in upper esophageal in 34 cases,distal tracheoesophageal fistula in one case.CT examination indicated distal tracheoesophageal fistula and orificium fistulae in 14 cases.Conclusion Combination of X-ray and CT imaging results with clinical symptoms can aid early detection and typing of con-genital esophageal atresia,furthermore provides guidance for operation and finally improves the survival rate of infants.

Objective To compare the tannin contents of Platycladus orientalis(L.) Franco and its carbonisatus.Method A tungsten molybdophosphate-casein colorimetric method was used with gallic acid as reference substance.Results The standard curve in the range of 0.026～0.26 mg(r =0.999 4),and the average recovery rate was 97.85%,RSD=1.07%(n =9).Conclusion The method is reliable and can be used to determine the tannic of and its different processed products.

Acute and chronic graft rejection are the major factors leading to graft non-function.There is an active expression of chemokines early after transplantation.They recruit T cells and antigen presenting cells selectively to the graft, leading to inflammatory reaction and finally to graft non-function.Accordingly,monitoring the expression status of chemokines and their receptors regularly may help to the diagnose rejection.To determine one or more chemokines or their receptors as the new targets for anti-rejection therapy will be of great clinical significance.This review focuses on the research progression in the above areas.