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.

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.

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 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 assess the efficacy and safety of recombinant human thrombopoietin (rhTPO) on chemotherapy-induced thrombocytopenia in patients with solid tumor.

METHODSIn this randomized crossover self-controlled multi-center clinical trial, 154 patients with solid tumor were randomly divided into two groups (group A 77 cases and group B 77 cases). All patients were given the same two cycles of chemotherapy. In group A, the first cycle was treated cycle, in which patients were given rhTPO, while the second cycle was non-treated cycle as a control. In group B, the first cycle was non-treated cycle as a control, while the second cycle was treated cycle. RhTPO 1.0 microg/(kg x d) was administered subcutaneously 6-24 hours after chemotherapy for the longest 14 days. Laboratory tests included complete blood counts, urinalysis, serum chemistry, coagulant test, chest radiography, and electrocardiogram. Serum samples were screened for anti-rhTPO antibodies.

RESULTSIn both group A and group B, platelet decrease and duration had no significant difference between the treated cycle and non-treated cycle. Platelet count was higher in the treated cycle, than in the non-treated cycle: [minimal mean platelet count (64.4 +/- 45.4) x 10(9) cells/L and (52.4 +/- 30.9) x 10(9) cells/L (P=0.000), maximal mean platelet count (263.9 +/- 142.5) x 10(9) cells/L and (148.9 +/- 67.7) x 10(9) cells/L (P=0.000)]. Duration of thrombocytopenia was shorter in the treated cycle than in the non-treated cycle [days with platelet count < 50 x 10(9) cells/L, (2.5 +/- 3.9) and (3.7 +/- 5.7) (P=0.04); days with platelet count recovered > or = 75 x 10(9) cells/L, (10.3 +/- 8.7) and (14.0 +/- 8.9) (P=0.000), and days with platelet count recovered > or = 100 x 10(9) cells/L, (15.9 +/- 10.5) and (21.1 +/- 9.5) (P=0.000)]. The need for platelet transfusion was not significantly reduced in treated cycle. The effects of rhTPO on WBC, Hb, hepatic function, renal function, and coagulant function were not found. Transient low-titer non-neutralizing antibody was developed in one patient. Therapy with rhTPO was tolerated by all patients. Mild side effects were observed in individual patients, including fever, dizziness, and chill. Conclusion Administration of rhTPO after chemotherapy can significantly reduce the degree and duration of thrombocytopenia and promote platelet recovery. Therapy with rhTPO seems to be safe.

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; adverse effects ; Cross-Over Studies ; Female ; Humans ; Lung Neoplasms ; blood ; drug therapy ; Male ; Middle Aged ; Neoplasms ; blood ; drug therapy ; Platelet Count ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Thrombocytopenia ; chemically induced ; drug therapy ; prevention & control ; Thrombopoietin ; therapeutic use

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

AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.

METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.

RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.

CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance

OBJECTIVETo screen the mutations of RET proto-oncogene in sporadic patients with pheochromocytoma.

METHODSForty-two cases of sporadic pheochromocytoma were tested for mutations of RET gene. Of these 42 DNA samples, 12 were extracted from peripheral blood cells and 30 from paraffin-embedded pheochromocytoma specimens. The PCR product of exon 10 and exon 11 was used to molecular analysis of the RET proto-oncogene.

RESULTSAmong 42 patients, 2 were found to have RET gene mutations. One of mutations located at codon 634 (TGC>TAC) in exon 11 of RET proto-oncogene. Another one located at codon 632 (GAG>AAG).

CONCLUSIONSome patients with apparently sporadic pheochromacytoma were carrier of mutations, a routine genetic analysis for mutations of RET gene is indicated for these patients.

Adrenal Gland Neoplasms ; diagnosis ; genetics ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Female ; Genetic Predisposition to Disease ; genetics ; Genetic Testing ; Humans ; Male ; Middle Aged ; Mutation ; Pheochromocytoma ; diagnosis ; genetics ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-ret ; genetics

OBJECTIVETo investigate the influence of recombinant human growth hormone (rhGH) on body fluid compartments and water-sodium retention in severe burn patients.

METHODSThirty adult patients with severe burn were divided into treatment (T) and control (C) groups by block randomized design. Patients in both groups were subcutaneously injected with same amount of rhGH (12 IU/d) or isotonic saline during 7 - 21 post burn day (PBD). The total body water (TBW), intracellular water (ICW), extracellular water (ECW) were measured by bioelectrical impedance analysis (BIA) on 7, 14, 21 PBD. The 24 h urinary output of Na+ was determined by ion selective electrode method (ISE).

RESULTSThere were no significant difference in levels of TBW, ICW, ECW and 24 h urinary output of Na+ between two groups on 7, 14, 21 PBD (P > 0.05). No difference in results was found between groups at different time points (P > 0.05). After the data were analyzed, the level of TBW (36 +/- 6 L), ICW (21 +/- 4 L) on 21 PBD were evidently lower than those on 7 PBD (38 +/- 6 L, 23 +/- 7 L, P < 0.01).

CONCLUSIONThe level of ICW and TBW in severe burn patients decreased along with the time. Proper dosage of rhGH has no significant effect on body fluid compartments and water-sodium retention.

Adolescent ; Adult ; Aged ; Body Fluid Compartments ; Body Water ; Burns ; metabolism ; physiopathology ; therapy ; Edema ; etiology ; Electric Impedance ; Extracellular Space ; Female ; Human Growth Hormone ; therapeutic use ; Humans ; Male ; Middle Aged ; Sodium ; metabolism ; Young Adult