Objective Multimeric cyclic RGD (Arg-Gly-Asp) peptides are capable of improving the integrin αvβ3-binding affinity due to the polyvalence effect.In this study,the authors prepare 99Tcm-la-bearing cyclic RGD tetramer E{E[c(RGDfK)]2}2,and evaluate its biodistribution and imaging in nude mice beating U87 MG human glioma xenografts with integrinαvβ3-positive.Methods 99Tcm-hydrazino-nictinamide (HYNIC)-E{E[c(RGDfK)]2}2 was prepared by two-step method,while HYNIC wag chosen as bifunctional chelator,and tricine and trisodium triphenylphosphine-3,3,3-trisuifonate (TPPTS) as coligands.The af-finity of c (RGDyK) monomer,HYNIC-E[c(RGDfK)]2 dimer and HYNIC-E{E[c(RGDfK)]2}2 tetramer to integrin αvβ3 was compared by in vitro competitive assay against binding of 125I-c(RGDyK)to integrin αvβ3.positive U87 MG human glioma cells.The biodistribution [the percentage of injection dose per gram of tissue(%ID/g)] and imaging were performed in nude mice bearing UB7MG human glioma xenografts.Re-suits The labeling yield of 99Tcm-HYNIC-E{E[c(RGDfK)2}2 was over 95%,and the radiochemical purity was more than 99%after purification with Sop-Pak C18 cartridge.The 50%inhibiting concentration (IC30) val-ues of c(RGDyk),HYNIC-E[c(RGDfK)]2 and HYNIC-E{E[c(RGDfK)]2}2 were 85.9,9.5 and 4.5 nmol/L, respectively.The result indicated that RGD tetramer possessed a significantly higher affinity to in-tegrinαvβ3.The biodistribution data showed that 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was excreted mainly through kidneys.The tumor uptake of 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was two times higher than 99Tcm- HYNIC-E[c(RGDfK)]2,at 1h postinjection,with the uptake of(10.32±0.07)%ID/g and(5.15±0.52)%ID/g,respectively,which was consistent with the in vitro competitive binding data.The tumor up-tale of 99Tcm-HYNIC.E{E[c(RGDfK)]2}2 was still as higher as(9.35±1.35)%ID/g at 4 h postinjec-tion, which demonstrated that the retention time of radiotracer in tumor was long enough.The imaging showed that tumor was clearly visualized at 1h postinjection,and the image at 4 h postinjection Was better.Conclusion The higher tumor uptake and longer retention time in tumor make 99Tem-HYNIC-E{E[c(RG-DfK)J 2}2 a promising radiotracer for integrinαvβ3-positive tumors imaging,furthermore,suggest that radi-onuelides(such as 90Y).1abeled RGD tetramer is more suitable for the therapy of integrin αvβ3-positive tumors.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Objective To report 6 cases of hydroa vacciniforme-like cutaneous lymphoma,and to inves- tigate the relationship between this disorder and Epstein-Barr virus(EBV)infection.Methods Pathological and immunohistochemical examinations were performed in the biopsy specimens obtained from all 6 patients. Skin lesions were subjected to EBV encoded RNA(EBER)detection by in situ hybridization.Serological assay and quantification of EBV DNA were performed.Results All the 6 patients had recurrent papules, papulovesicles,necrosis and variola-like scar with chronic intermittent fever;four of the patients also presented with edema of the face,hands and feet.Pathologically,there were multilocular vesicles in the epidermis,and large numbers of infiltrating lymphocytes through the dermis.The cells were atypical with mitotic figures. Immunohistochemical staining of the lesions of 4 patients showed large quantities of cells expressing CD56, scattered cells expressing CD3 and CD45RO,and cells expressing grazyme B and T cell intracellular antigen-1 (TIA-1);a diagnosis of hydroa vacciniforme-like cutaneous NK/T lymphoma was made in these 4 cases. In the lesions of another 2 patients,the cells expressing CD3 and CD45RO,but not CD56,were observed; the diagnosis of hydroa vacciniforme-like cutaneous T-cell lymphoma was made in them.EBER was detected in the tumor cells of all the 6 patients.The IgG titers of anti-Epstein-Barr viral capsid antigen increased in all patients(1:5120 in 2 cases,1:2560 in 2 cases,1:1280 in 2 cases).The copies of EBV DNA were increased in the peripheral blood of both the two detected cases.A chronic active EBV infection was confirmed in all patients.Conclusions Hydroa vacciniforme-like cutaneous lymphoma is clinically characterized by edema of face,hands and feet,vesicular eruptions and variola like scars;histologically,it is characterized by infiltrates of atypical cells consistent with lymphoma,and necrosis in the center of vessels.NK/T is the primary immunophenotype of this disease.There is a close association between chronic active EBV infection and hydroa vacciniforme-like cutaneous lymphoma.

OBJECTIVE: To research the expression of alternative splicing segment of fibronectin-IIIcs in incised wound of skin, to offer the experimental data for early injury time judgement. METHODS: Using in situ hybridization with DIG-labeled anti-sense RNA probe, the expression of FN-IIIcs domain was detected in both human and rat skin incised wound. RESULTS: (1) The expression of FN-IIIcs improved from the time of 30 min after injury, reached peak at 6 h and then reduced gradually. (2) The positive expression cells were mainly distributed in hair follicles, sebaceous glands and endothelium, however, in human beings they were basement cells of epidermis. CONCLUSION The expression of FN-IIIcs domain would be a supposed useful criteria for early injury time judgement.
Animals ; Fibronectins/genetics* ; Forensic Medicine ; Humans ; Male ; RNA, Messenger/biosynthesis* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Time Factors ; Wound Healing/physiology*

OBJECTIVE: To analyze the polymorphism of rs220030, a SNP which is located in the promoter region of small nuclear ribonucleoprotein polypeptide N (SNRPN) gene in the Chinese Han population and to obtain the data of population genetics. METHODS: The denaturing gradient gel electrophoresis (DGGE) method was applied to detect the polymorphism of rs220030 in 100 unrelated and healthy individuals from the Shanghai Han population. The genotyping result of this SNP was confirmed by TaqMan assay in some typical samples. RESULTS: DGGE results showed 4 bands for CT heterozygote, and 1 band for CC or TT homozygote, and those results were confirmed by The TaqMan SNP genotyping assays. Genotyping results showed 34 individuals with CC, 41 with CT and 25 with TT of rs220030. The allele frequencies for C and T were 0.545 and 0.455, respectively. H was 0.500, PIC was 0.373, DP was 0.654, and PE was 0.186. The distribution of genotype frequencies were in Hardy-Weinberg equilibrium. CONCLUSION DGGE is a quick and effective method in the analysis of SNP polymorphism in small population. Statistical parameters of rs220030 for forensic evaluation meet the requirements for forensic identification and paternity testing.
Alleles ; Asian People/genetics* ; China/ethnology* ; DNA Primers ; Denaturing Gradient Gel Electrophoresis/methods* ; Gene Frequency ; Genetic Markers ; Genetics, Population ; Genotype ; Heterozygote ; Humans ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide/genetics* ; Promoter Regions, Genetic ; snRNP Core Proteins/genetics*

OBJECTIVE: To explore the relationship between the expression of EIIIA+ fibronectin in incised wound of rat's skin and injury time. METHODS: The wounding model was established by cutting the dorsal skin of 48 adult SD rats. The rats were sacrificed at the pre-set injury time as immediately, 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, and 8 h. The skin samples were taken at the margin of wound. The expression of the EIIIA? fibronectin was detected by immunohistochemistry and Western blotting and the relationship be- tween its expression and injury time was observed. Results The expression of EIIIA+ fibronectin was not observed immediately. The basal cell of skin began to show positive expression 0.5 h after injury. With the extension of injury time, positive staining became stronger. The value of relative optical density was gradually increased with prolonged injury time by the Western blotting analysis. CONCLUSION The expression of EIIIA+ fibronectin could be used for estimation of injury time in the early stage of skin injury.
Animals ; Fibronectins/metabolism* ; Immunohistochemistry ; Proteins/metabolism* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Staining and Labeling ; Time Factors

To investigate the chemical constituents of the whole plants of Bidens bipinnata, the separation and purification of constituents were performed by chromatography on macroporous resin, silica gel, MCI and Sephadex LH-20. Their structures were elucidated by spectroscopic data as quercetin (1), quercetin-3-0-alpha-L-rhamnoside (2), keampferol-3-O-beta-D-glucopyranoside (3), keampferol-3-O-alpha-L-rhamnoside (4), 3', 5-dyhydroxy-3, 6, 4'-trimethoxyl -7-O-beta-D-glucopyranoside flavonoid (5), 7, 8, 3', 4'-tetraflavanone(6), (2S)- and (2R)-isookanin-7-O-beta-D- glucopyranoside (7a/7b), (2S)- and (2R)-3'-methoxy-isookanin-8-O-beta-D-glucopyranoside (8a/8b), 6, 7, 3', 4'-tetrahydroxyaurone(9), maritimetin (10), esculetin (11), 3-O-caffeoyl-2-methyl-d-erythrono-1, 4-lactone (12), (7S, 8R) balanophonin-4-O-beta-D-glucopyranoside (13), eugenyl-O-beta-apiofuranosyl-( 1"-6') -O-beta-glucopyranoside (14), and (+)-syringaresinol-4'-O-beta-D-glucopyranoside (15). Compounds 8, 13, 14, and 15 were isolated from this genus for the first time. Compounds 1 and 6 were potent inhibitors against HSC-T6 cells in vitro and compounds 1, 2, 6, and 7 were capable of decreasing the inflammatory cytokine production of macrophage cells in vitro.
Bidens ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVE: To estimate the effort between antisense RNA probe and random primed DNA probe. METHODS: The in situ hybridization was conducted on parafin section from wounding model of rat skin. RESULTS: Although both probes appeared positive staining, RNA probes was superior to DNA probes in terms of depth of staining and background. CONCLUSION RNA probe showed more satisfactorily on ISH.
Animals ; DNA, Antisense ; Digoxigenin ; Fibronectins/genetics* ; In Situ Hybridization/methods* ; RNA Probes ; RNA, Messenger/biosynthesis* ; Random Amplified Polymorphic DNA Technique/methods* ; Rats ; Rats, Sprague-Dawley ; Skin/pathology*

OBJECTIVE: To investigate the signification of CTnI in acute myocardial infarction. METHODS: The model of myocardial infarction was established by ligating the left ventricle branch. Immunohistochemistry and image analysis were used to detect the change of CTnI in the model, and The sensitivity of immunohistochemistry and HE coloration were also compared. RESULTS: The acute myocardial infarction tissue showed obvious depletion of CTnI, there was no characterization of myocardial infarction in HE coloration. CONCLUSION CTnI is sensitive to diagnosis of acute myocardial infarction.
Animals ; Antibodies, Monoclonal ; Disease Models, Animal ; Female ; Immunohistochemistry ; Male ; Myocardial Infarction/metabolism* ; Myocardial Ischemia/pathology* ; Myocardium/pathology* ; Rabbits ; Sensitivity and Specificity ; Time Factors ; Troponin I/metabolism*

OBJECTIVEThe aim of the study was to review the cases of Kawasaki Disease (KD) and analyze the clinical features especially their cardiac complications.

METHODSTotally 283 patients with KD were hospitalized from 1992 to 2002. Their clinical features and factors associated with increased risk of coronary artery aneurysms were reviewed.

RESULTS(1) Among the 283 KD patients, 186 were male and 97 were female. The male-female ratio was 1.9:1. Most of them (71%) were younger than 3 years old. Seasonal peak was in spring and summer (from May to Aug). Depending on the criteria of KD, 228 (81%) were diagnosed as typical KD and 55 (19.4%) were atypical KD. All patients had fever, lasting for 6.1 days. The most common clinical features were oral mucosal changes (97.5%) and cervical lymphadenopathy (95.4%), conjunctivitis (91.2%). And changes in the extremities (89.8%) and rash (81.5%) were also noted. (2) Before the treatment, coronary artery abnormalities were seen in 103/279 (36.9%), which occurred within 4 - 30 days of fever onset. Two weeks after intravenous gamma globulin (IVIG) treatment, the new cases of coronary artery abnormalities were 28/211 (13.3%). The prevalence of coronary artery aneurysms (CAA) with KD was 4.7%. The risk factors of CAA were male cases (P < 0.05) and fever lasting longer than 9 days (P < 0.05). Other cardiac abnormalities in acute phase included left atrial and ventricular enlargement (40/279, 14.3%) and changes in ECG (57/274, 20.8%). The pericardial effusions were found in 11 cases (3.9%).

CONCLUSIONSCardiac complications of KD occurred in the early period of KD. The new cases of coronary artery abnormalities were 13.3% after IVIG treatment. The risk factors of CAA included male cases and fever lasting for longer time.

Child, Preschool ; Coronary Aneurysm ; epidemiology ; etiology ; Female ; Heart Diseases ; diagnosis ; etiology ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; diagnosis ; drug therapy ; Prevalence ; Risk Factors