BACKGROUND:Sensitivity of diagnoses differentiating smaller nodes of splenic metastasis (＜ 1 cm) from CT and MRI is poor. So whether superparamagnetic iron oxide can enhance magnetic resonance imaging (MRI) of splenic VX2 tumor in rats need to be further studied.OBJECTTVE: To establish splenic VX2 tumor models, investigate MRI scanning combining with superparamagnetic iron oxide of specific reticuloendothelial system, and study the diagnostic significance of superparamagnetic iron oxide-enhanced MR images on splenic metastases.DESIGN: Duplicated-measured animal study.SETTING: Medical Imaging Center, Nanfang Hospital of the Southern Medical University.MATERIALS: The experiment was carried out in the Medical Imaging Center (Military Key Laboratory), Nanfang Hospital of the Southern Medical University from May 2005 to March 2006. A total of 25 adult SD rats, of either gender, weighing 200-300 g, were selected in this study. The animal experiment had got confirmed consent from local ethic committee. All rats were randomly divided into tumor group (n =20) and blank control group (n =5).METHODS: Models of VX2 tumor in spleen were established successfully. The images obtained before and after administration of superparamagnetic iron oxide. T1-weighted image(T1WI) (450/12 ms) and T2-weighted image(T2WI)(4 000/128 ms) were used to scan sequences. The imaging parameters of various tissues were analyzed before and after superparamagnetic iron oxide-enhanced MRI scanning. Rats in the blank control group were not used to establish models and the sequences were scanned as the same as those in the experimental group.MAIN OUTCOME MEASURES: Signal characteristics of various tissues before and after before and after superparamagnetic iron oxide-enhanced MRI scanning.RESULTS: Eight out of 20 rats died in the tumor group and other 12 and 5 rats in the control group were involved in the final analysis. MR scanning indicated that 8 rats in the tumor group had splenic VX2 tumor. ① On plain MR scanning T1-weighted splenic VX2 tumor showed hypointensity or isointensity which approximated to the signal intensity (SI) of splenic parenchyma. All lesions had been not displayed well. ② On superparamagnetic iron oxide enhancement T2WI sequence the SI of splenic parachyma decreased obviously with percentage of signal intensity loss (PSIL) was 56.11%,But the SI of tumor was not evidently change with PSlL was 1.90%. Nevertheless the signal-to-noise ratio (SNR) of normal splenic parenchyma around the lesions had obviously difference (P ＜ 0.001) comparatively. So the contrast between tumor and spleen increased, and tumor displayed more clear. Moreover the contrast-to-noise (CNR) between VX2 tumor and splenic parenchyma had an evidently difference with before and after admininstration of superparamagnetic iron oxide (P ＜ 0.001 ).CONCLUSION: On superparamagnetic iron oxide enhancement T1WI sequence the contrast of tumor-to-spleen is poor.So it is not good to characterize the lesions in spleen. On superparamagnetic iron oxide enhanced T2WI the contrast degree of lesions increased obviously. Consequently, superparamagnetic iron oxide-enhanced T2WI MRI scanning can improve the rate of detection and characterization in lesions of spleen.

Aim To search for colchicine derivatives which have high efficacy and low toxicity. Methods Colchicine was firstly converted into thiocolchicine, and then it was hydrolyzed to get 7-(N-deacetylthiocolchicine). At last, 7-(N-deacetylthiocolchicine) was amidated to get the target compounds. The chemical structure of these new derivatives was confirmed with 1H NMR, IR, MS, and HR-MS. The cytotoxicity of the compounds was tested by MTT assay. Their in vivo antitumor activity was evaluated against mice tumor H22 and U14. Results Twelve thiocolchicine derivatives are new compounds. Conclusion In vitro antitumor activity has showed that some of these thiocolchicines possessed cytotoxic activity superior to colchicine. However, in vivo antitumor activity indicated that these derivatives have poor efficacy in mice.

Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.

Objective To evaluate the efficiency of BIOMED-2 system in detecting IGH gene clonal rearrangement and application in diagnosis of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) lymphoma. Methods Forty-five cases were collected, including 36 MALT lymphomas from different organs, 3 extranodal lymphoid tissue proliferative lesions and 6 severe gastritis associated with H pylori. DNA was extracted from the formalin fixed paraffin embedded blocks of these cases and the quality of DNA was assessed using the BIOMED-2 specimen mixed control primers. IGH gene clonal rearrangement was detected using IGH VH-JH primers. The sensitivity and specificity of BIOMED-2 PCR were analyzed.Results Adequate DNA (≥ 300 bp) was obtained in 31 of 45 samples (including 22 MALT lymphomas, 3 lymphoid tissue proliferative lesions and 6 severe gastritis), and the DNA from the other 14 samples was degraded seriously. 16 of 22 MALT lymphomas were positive of IGH clonal rearrangement with the sensitivity of 72.7 ％.In contrast,none of 6 severe gastritis was positive with the specificity of 100 ％ IGH and clonal rearrangement were detected in one of 3 lymphoid tissue proliferative lesions. Conclusion BIOMED-2 assay is an effient and reliable method for diagnosis and differential diagnosis of MALT lymphoma which is important for clinical practical value.

Objective To evaluate the diagnostic value of autoantibodies to cell membrane associated with DNA (mDNA) in systemic lupus erythematosus (SLE) and the combination with other autoantibodies in the diagnosis of SLE. Method The anti-mDNA antibody had the characteristic pattern of perip-heral membrane fluorescence on cultured HL60. The same serum samples were detected for other antibo-dies of SLE. Pearson's Chi-square test was used for statistical analysis. Results This pattern was observed in 145 of 205 serum samples of SLE patients , but in 5 of 55 the serum samples of rheumatoid arthritis , in 10 of 45 primary Sjogren syndrome's patients and in 4 of 35 PM/DM and absent in 50 blood donors. The sensitivity and specificity of anti-mDNA antibody to SLE was 70.7% and 86.7%. The sensitivity and specificity of combined anti-mDNA antibody and ANA was 94.6% and 76.7%. The sensitivity and specificity of combined anti-mDNA antibody and anti-dsDNA antibody was 76.8% and 95.5%. The sensitivity and specificity of combined anti-mDNA antibody and anti-Sm antibody was 79.6% and 100%. The sensitivity and specificity of combined anti-mDNA antibody and AnuA was 93.0% and 100%. Conclusion This novel rapid immunofluorescence method can be a useful diagnostic test for SLE patients. Due to its high sensitivity and specificity, it is better than other diagnostic tests such as anti-dsDNA antibody and anti-Sm antibody for the diagnosis of SLE.

To establish a rat model of rheumatoid arthritis (RA) with kidney deficiency syndrome.

One hundred children aged 8-10 years and 100 adults aged 18-45 from their families in three townships Baoshan,Mingcheng,and Yantongshan of Panshi city,Jilin province were selected in April 2009. In addition,50 pregnant or lactating women,and their 50 infants aged 0-2 years were selected.The median level of iodine in salt at household was 30.2 mg/kg and that of drinking water was 2.8 μg/L.The median levels of urine iodine in school age chidren ( 196.5 μg/L),during pregnancy( 198.5 μag/L),lactation( 224.9 μg/L),and in infants (209.0 μg/L) all were optimal according to World Health Organization criteria.Only the median urine iodine in adults ( 269.0 μg/L) was more than adequate.The prevalence of goiter evaluated by B-ultrasound was 1.8％ in 8-10year-old children.The incidences of various thyroid dysfunctions were low in these residents,such as subclinical hyperthyroidism ( 2.1％ ),subclinical hypothyroidism ( 1.5％ ),and hyperthyroidism ( 1.2％ ). Subclinical hypothyroidism occurred mostly in adults (2.7％),pregnant women ( 1.7％ ),and lactating women ( 1.8％ ) ; and subclinical hyperthyroidism occurred mostly in 8-10 years old school chidren (4.5％) and lactating women (3.6％).These results show that the current iodized salt content has no harmful effect on thyroid function of rural residents in Jilin province.

Three cases of swimming pool granuloma are reported.Case 1:a 40-year-old female presented with a 2-month history of nodules and plaques on the right hand and forearm.She was a tropical fish salesperson but denied trauma history.Skin examination revealed multiple irregularly sized,dark-red nodules and plaques on the joints of right fingers,wrist,and elbow,as well as multiple subcutaneous nodules simulating strings of beads on the right upper limb.Case 2:a 48-year-old female presented with a 2-month history of nodules and plaques on the left hand and forearm.There was a history of trauma due to tropical fish tank and filter cleaning.Physical examination showed multiple deep purple plaques and painless subcutaneous nodules scattered on the left hand,wrist,and upper limb.Case 3:a 39-year-old male presented with a 3-month history of nodules on the fingers of both hands.There was no history of trauma,but he was a tropical aquarist.Skin examination revealed multiple soybean-sized dark-red nodules on the extensor aspect of interphalangeal joints of both hands.Fungal examinations yielded negative results in the 3 cases,while histopathology revealed infectious granuloma with a mixed inflammatory cell infiltrate.All of the cases showed positive results in purified protein derivative (PPD)skin test.Mycobacterium marinum was isolated from the lesional tissue of Case 1 and 2,but not from Case 3.All the patients were diagnosed with swimming pool granuloma,and given anti-atypical mycobacterial therapy including oral rifampin and clarithromycin.The lesions disappeared after 1 to 3 months of treatment,with the treatment course varying from 2 to 5 months.No recurrence was observed during a 3- to 12-month follow-up.

Objective To investigate the role of nuclear transcription factor kappa B (NF-κB) -matrix metalloproteinase-9 (MMP-9) signaling pathway in delayed encephalopathy after acute carbon monoxide poisoning (DEACMP). Methods 150 male SD rats were randomly assigned to air control group (AC group) , CO poisoning group (CO group) , pyrrolidine thiocarbamate (PDTC) + CO poisoning group (PC group). DEACMP model was reconstructed by modified intraperitoneal injections. The 1, 3, 7, 14, and 21 d after intraperitoneal injection were observed here by different approaches. Morris water maze test was used to test the learning and memory ability of rats.HE staining was used to observe the morphology of hippocampal CA3 cells. Immunofluorescence and Western Blot methods were used to detect the expression of NF-κB and MMP-9. RT-PCR was used to measure the expression of MMP-9 mRN A. Transmission electron microscopy was used to observe the ultrastructure of synapses. Results After14 days, the average intubation period of CO group was longer than that of AC group (P < 0.05) , and that of PC group was shorter than that of CO group (P < 0.05). However, average intubation period of PC group was longer than that of AC group (P< 0.05). In CO group, the expression of NF-κB in hippocampus increased (day 1). At day 3, the expression of NF-κB rapidly increased. The expression of MMP-9 gene and protein increased in the first three days and then decreased thereafter. The expression of NF-κB and MMP-9 in PC group was lower than that in CO group (P < 0.05) , while it was higher than AC group (P < 0.05). The peak value of apoptosis in CO group was delayed to 7-14 d after exposure, the apoptotic cells in PC group decreased significantly, and it was obvious on the 14 th day.Electron microscopy showed that the damage of synapses ultrastructure in CO group was significantly heavier than that in PC group on the 14 th day. Conclusions NF-κB-MMP-9 signal pathway leads to DEACMP, and PDTC could alleviate the impairment of learning and memory ability in rats with acute CO poisoning.

OBJECTIVETo clone and identify the gene encoding human ubiquitin binding enzyme 2 and study its expression pattern.

METHODSAccording to the sequence of human EST, which is highly homologous to the mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformatics technique and its expression pattern was studied by using multiple-tissue Northern blot.

RESULTSTwo cDNA clones encoding human ubiquitin conjugating enzyme have been isolated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows that they are expressed exclusively in adult human heart, placenta, and pancreas but no transcripts can be detected in brain, lung, liver, skeletal muscle or kidney.

CONCLUSIONSThe gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post-translation.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Female ; Humans ; Mice ; Molecular Sequence Data ; Myocardium ; metabolism ; Pancreas ; metabolism ; Placenta ; metabolism ; Rats ; Sequence Alignment ; Sequence Analysis, DNA ; Ubiquitin ; genetics ; Ubiquitin-Conjugating Enzymes ; biosynthesis ; chemistry ; genetics