Objective: To observe mRNA and protein expression of leukotriene B4 (LTB4) receptor 2 (BLT2)in mice during the course of development from colitis to colitis-associated cancer. Methods: ICR mice were used to establish the animal model of colitis-associated colon cancer and randomly divided into control group and experimental group. We began to administer inducer on d0. Mice were sacrificed at 2, 3, 5, 7, 9, 13, and 18w, respectively. Histopathological changes in colons were examined. The protein and mRNA expression of BLT2 in colons were measured by immunohistochemical assay and real-time quantitative PCR, respectively. Results: Histological study showed that with the extension of time, the lesions of mice colons aggravated, first a mild inflammation, then atypical hyperplasia, and finally to colon cancer. Immunohistochemical staining showed that the expression of BLT2 protein was low in normal colon, but high in inflammatory lesions, especially in inflammatory cells. There was no BLT2 expression in atypical hyperplasia and cancer cells, while BLT2 was highly expressed in the stroma and lumans of cancer tissue. Compared with the control group, mRNA expression of BLT2 in the experimental group was significantly higher at 2, 13 and 18w (P<0.05); and very significantly higher at 3, 5, 7 and 9w(P<0.01). Conclusion: The mouse model of colitis-associated colon cancer was developed in this study. The expression of BLT2 changed in the course of colitis development, indicating that BLT2 may play an important role in the transition process from colitis to colon cancer.

Pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes, which "chondron" with its enclosed cells. A number of studies suggested that PCM is rich in proteoglycans, collagen and fibronectin, and plays an important role in regulating microenvironment of chondrocytes. Direct measures of PCM properties through micropipette aspiration technique showed that PCM was different from mechanical property of chondrocytes and nature extracellular matrix. However, the function of PCM is not clear, and need further study.
Animals ; Biomechanical Phenomena ; Chondrocytes ; chemistry ; cytology ; metabolism ; Extracellular Matrix ; chemistry ; metabolism ; Humans

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Objective To explore the effect of atorvastatin on cardiac hypertrophy and to determine the potential mechanism involved. Methods Anin vitro cardiomyocyte hypertrophy from neonatal rats was induced with angiotensinⅡ (AngⅡ) stimulation. Before AngⅡ stimulation, the cultured rat cardiac myocytes were pretreated with atorvastatin at different concentrations (0.1, 1, and 10μmol/L). The following parameters were evaluated: the myocyte surface area,3H-leucine incorporation into myocytes, mRNA expressions of atrial natriuretic peptide, brain natriuretic peptide, matrix metalloproteinase 9, matrix metalloproteinase 2, and interleukin-1β, mRNA and protein expressions of theδ/β peroxisome proliferator-activated receptor (PPAR) subtypes. Results It was shown that atorvastatin could ameliorate AngⅡ-induced neonatal cardiomyocyte hypertrophy in the area of cardiomyocytes,3H-leucine incorporation, and the expression of atrial natriuretic peptide and brain natriuretic peptide markedly. Meanwhile, atorvastatin also inhibited the augmented mRNA level of several cytokines in hypertrophic myocytes. Furthermore, the down-regulated expression of PPAR-δ/β at both the mRNA and protein levels in hypertrophic myocytes could be significantly reversed by atorvastatin treatment. Conclusions Atorvastatin could improve AngⅡ-induced cardiac hypertrophy and inhibit the expression of cytokines. Such effect might be partly achieved through activation of the PPAR-δ/β pathway.

Objective To analyze the classification,MR manifestations,and the pathological basis of solitary necrotic nodule of the liver(SNN)in order to evaluate MRI as a diagnosing tool Methods The MR appearances of 9 cases with pathologically proved SNN were analyzed and correlated with the classification and pathological appearances.Relevant literature was reviewed.Results(1)Simple coagulative necrosis type(5 cases):The signal of lesions was hypo-intense or iso-intense on both T_1-and T_2- weighted images.After Gd-DTPA administration,the internal part of the lesions showed no enhancement,while the thin capsule of the lesions demonstrated mild or moderate delayed enhancement. These lesions,proved by pathology,were composed of central coagulative necrotic core and a peripheral hyaline fibrosis capsule.(2)Coagulative necrosis aceompanied by liquefactive necrosis type(1 case):On T_1-weighted images,the signal of hypo-intensity was found within these lesions and even lower signal intensity was found in the central area of larger lesions.On T_2-weighted images,the lesions had a bright core and a peripheral hypointensive or isointensive area.After Gd-DTPA administration,the internal part of the lesions showed no enhancement,while the thin capsule of the lesions demonstrated mild or moderate delayed enhancement.These lesions had a central coagulative necrosis core interleaved by slit- like liquefactive necrosis foci,and peripherally a thin capsule of hyaline fibrosis proved by pathology.(3)Multi-nodular fusion type,(3cases):On T_1-weighted images,the lesions were of hypointensive or isointensive signal and had multiple septa of isointensive signal.On T_2-weighted images,the lesions were of hypointensive or isointensive signal and had multiple septa of hyperintensive or isointensive signal.After Gd-DTPA administration,No enhancement was found except mild or moderate delayed enhancement found in the thin capsule and septa.These lesions were composed of central coagulative necrosis area and a peripheral hyaline fibrosis capsule with multiple internal septa proved by pathology.Conclusion MRI apperances can reflect the classification and pathological features of solitary necrotic nodule of the liver.

As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cricetinae ; Cricetulus ; Gene Expression Profiling ; Green Fluorescent Proteins ; genetics ; Humans ; Molecular Sequence Data ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Tissue Distribution ; Transfection

OBJECTIVETo study the genetic association of apolipoprotein (apo) E and apoCI gene polymorphisms with coronary heart disease (CHD) in China.

METHODSapoE genotypes were identified by multiplex amplification refractory mutation system (multi-ARMS) and the apoCI promoter polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 186 cases with CHD (age: 65.0 +/- 10.5 years) and 350 controls (age: 63.6 +/- 8.3 years). The haplotype frequencies were estimated.

RESULTSThe frequencies of apoE E4/3 genotype (26.9%) and epsilon4 (14.5%) in CHD group were significantly higher than that in the control group (12.6%, 7.0%), P <0.05. The significant difference was also found for the apoCI locus and the CHD group showed higher rate of both for the H2 allele and genotypes, carrying this allele. Estimation of the haplotype frequencies indicated that the association between the apoE-CI haplotype and CHD was significantly strong. The apoE-epsilon4/apoCI-H2 was estimated to be responsible for 9.86% of CHD.

CONCLUSIONWhen the subjects carrying both epsilon4 and H2 alleles, they would have higher risk of suffering from CHD than controls.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Apolipoproteins C ; genetics ; Apolipoproteins E ; genetics ; China ; epidemiology ; Coronary Disease ; blood ; epidemiology ; genetics ; Female ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Risk Factors

Adult ; Dioxins ; Glutathione Peroxidase ; blood ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; blood ; Middle Aged ; Occupational Exposure ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; blood

BACKGROUNDThe use of a free, vascularized fibular graft is an important technique for the reconstruction of large defects in long bones. The technique has many advantages in strong, tubular bones; a more reliable vascular anatomy with a large vascular diameter and long pedicle is used, minimizing donor-site morbidity. Due to limitations in both fibular anatomy and mechanics, they cannot effectively be used to treat large limb bone defects due to their volume and strength.

METHODSFrom 1990 to 2001, 16 clinical cases of large bone defects were treated using vascularized double-barrel fibular grafts. Patients were evaluated for an average of 10 months after surgery.

RESULTSAll the patients achieved bony union; the average bone union took 10 months post surgery, and no stress fractures occurred. Compared with single fibular grafts, the vascularized double-barrel fibular grafts greatly facilitate bony union and are associated with fewer complications, suggesting that the vascularized double-barrel fibular graft is a valuable procedure for the correction of large bone defects in large, long bones in addition to enhancing bone intensity.

CONCLUSIONSThe vascularized double-barrel fibular graft is superior to the single fibular graft in stimulating osteogenous activity and biological mechanics for the correction of very large bone defects in large, long bones. Free vascularized folded double-barrel fibular grafts can not only fill up large bone defects, but also improve the intensity margin. Therefore, this study also widens its application and enlarges the treatment targets. However, in the case of bone deformability, special attention should be paid to bone fixation and protection of donor and recipient sites.

Adolescent ; Adult ; Bone Diseases ; pathology ; surgery ; Bone Transplantation ; methods ; Female ; Fibula ; pathology ; surgery ; Humans ; Lower Extremity ; pathology ; surgery ; Male ; Middle Aged ; Models, Biological ; Reconstructive Surgical Procedures ; methods ; Reproducibility of Results

Neutrophils play a major role in host defense against microbial infection. There are some clues indicate that neutrophils may also play a role in the pathophysiology of the airway obstruction in chronic asthma. We studied the roles of intracellular calcium and GTP gamma S in the regulation of neutrophils exocytosis using pipette perfusion and membrane capacitance measurement technique in whole cell patch clamp configuration. The results showed that the membrane capacitance increase induced by calcium revealed a biphasic process. The first phase occurred when the calcium level was between 0.2-14 micromol/L with a plateau amplitude of 1.23 pF and a calcium EC50 of 1.1 micromol/L. This phase might correspond to the release of the tertiary granules. The second phase occurred when the calcium concentration was between 20-70 micromol/L with a plateau increment of 6.36 pF, the calcium EC50 being about 33 micromol/L. This phase might represent the release of the primary and secondary granules. Intracellular calcium also simultaneously increased the exocytotic rate and the eventual extent in neutrophils. On the other hand, GTP gamma S can increase the exocytotic rate in a dose-dependent manner but had no effect on the eventual extent of membrane capacitance increment (>6 pF) if the cell was stimulated for a long period (>20 min). GTP gamma S (ranging from 20 to 100 micromol/L) induced the neutrophils to release all four types of the granules at very low intracellular calcium level.
Calcium ; metabolism ; Cell Degranulation ; drug effects ; Exocytosis ; drug effects ; GTP-Binding Proteins ; metabolism ; physiology ; Guanosine Triphosphate ; analogs & derivatives ; pharmacology ; Humans ; Neutrophils ; metabolism ; physiology ; Patch-Clamp Techniques