Objective To compare the different response of dermal fibroblasts from normal and scar-ring skin to cytokines,extracellular matrix and collagen promoter.Methods Human dermal fibroblast cul-ture was established by explanting tissue specimens from normal and scarring skin.Cellular proliferative ac-tivities were determined with BrdU-ELISA after treatment with IFN-?A,hIFN-?,TNF?and laminin for24hours.Reporter genes driven by collagen promoters were analyzed in two types of dermal fibroblasts trans-fected with consensus constructs48hours later.Results Dermal fibroblasts from normal and scarring skins showed different proliferative responses to IFN-?A,hIFN-?,TNF?and laminin,with stimulatory effects pre-sented in normal fibroblasts treated with IFN-?A,hIFN-?,TNF?and laminin,inhibitory effects presented in scarring fibroblasts treated with IFN-?A and laminin,and reduced or resistant effects presented in scar-ring fibroblasts to hIFN-?and TNF?.The5'flanking sequences(-2483~+42bp,-1448~+42bp)from human?1(Ⅰ)procollagen gene had lower activity of promoter in scarring fibroblasts compared with that in normal fibroblasts.Conclusion Compared with normal dermal fibroblasts,scarring dermal fibroblasts have reduced response or are resistant to some cytokines.Lower collagen production capacity is anticipated since collagen promoter activity is low in scarring dermal fibroblasts.The pathological mechanism of scarring for-mation might be related to the change of responses of dermal fibroblasts.

OBJECTIVETo investigate clinical implications of expression of CD40L in monocytes and changes in serum soluble CD40L in patients with acute coronary syndromes (ACS).

METHODSSixteen control and 56 patients, including 24 with stable angina (SA), 20 with unstable angina (UA) and 12 with acute myocardial infarction (AMI) enrolled in this study. Expression of CD40L in monocytes was analyzed by flow cytometry and sCD40L levels were measured by ELISA.

RESULTSExpression of CD40L in monocytes and serum levels of sCD40L in UA and AMI patients were higher than in SA patients and controls. In patients with AMI, sCD40L levels showed no significant increase when compared to patients with UA, while AMI patients had a peak level of sCD40L at 24 hours after AMI. PTCA induced a marked rise in sCD40L levels in all patients, while CD40L expression in monocytes showed no difference between patients with PTCA, before and after.

CONCLUSIONEnhanced level of serum sCD40L may be a reliable prognostic indicator for ACS and may represent a marker of coronary disease activity.

Angina Pectoris ; blood ; pathology ; CD40 Ligand ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Monocytes ; metabolism ; Myocardial Infarction ; blood ; pathology

OBJECTIVETo study the expression of tenascin in normal and fibrotic rat liver.

METHODSLiver fibrosis induced in rat with CCl(4) were divided into three stages: the stage of hepatic injury (4 weeks), early stage of hepatic fibrosis (8 weeks) and later stage of hepatic fibrosis (12 weeks). Tenascin expression in liver tissue was observed by immunohistochemical method and in situ hybridization using digoxigenin-labelled DNA probe.

RESULTSIn normal rat liver there was a weak staining for tenascin. In both liver injury stage and early stage of hepatic fibrosis, both mRNA signal and immunostaining for tenascin were significantly increased as compared to that in normal liver. In later stage of hepatic fibrosis, mRNA signal and immunostaining for tenascin were decreased compared with that in early stage of hepatic fibrosis. The cellular source of tenascin in liver mainly restricted in mesenchymal cells.

CONCLUSIONSTenascin is a component of the extracellular matrix of liver tissue. Plays a role in early matrix organization during liver fibrogenesis.

Animals ; Carbon Tetrachloride ; Disease Models, Animal ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Image Processing, Computer-Assisted ; Immunohistochemistry ; In Situ Hybridization ; Liver Cirrhosis ; chemically induced ; metabolism ; Male ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Tenascin ; biosynthesis ; genetics

OBJECTIVETo evaluate the value of measurement of M2 autoantibodies in diagnosis of primary biliary cirrhosis (PBS).

METHODSM2 autoantibodies were detected by ELISA combined with western-blot.

RESULTSThirty-two cases presented positive M2 autoantibodies, including 20 cases of PBC, 9 liver diseases of unknown etiology, 1 autoimmune hepatitis and 1 drug-induced liver injury.

CONCLUSIONSM2 autoantibodies show good sensitivity and specificity to diagnosis of PBC. It should be used as a powerful and specific maker for the diagnosis of PBC

Autoantibodies ; immunology ; Autoantigens ; immunology ; Chemical and Drug Induced Liver Injury ; Enzyme-Linked Immunosorbent Assay ; Hepatitis, Autoimmune ; Humans ; Liver Cirrhosis, Biliary ; diagnosis ; Sensitivity and Specificity

AIM: To investigate the mechanism underlying insulin-stimulated increase in glucose uptake during low-flow myocardial ischemia. METHODS: The expression of myocardial GLUT1 polypeptide was determined by semiquantitative immunoblotting. The expression of GLUT1 mRNA was determined by semiquantitative Northern blotting. RESULTS: After infusing insulin during low- flow myocardial ischemia for 8 h，the expression of both GLUT1 mRNA and GLUT1 polypeptide was significantly higher in experimental myocardium than that in normal myocardium. The glucose uptake was upregulated at the same time in the exprimental myocardium. CONCLUSION: Insulin enhances the expression of GLUT1 mRNA and GLUT1 polypeptide in ischemic myocardial regions. GLUT1 expression may be an important mechanism by which myocardial cells enhance glucose uptake and metabolism during low-flow myocardial ischemia.

Objective: To study retrovirus (RV)-mediated transduction of gastric carcinoma cells with the herpes simplex virus thymidine kinase (HSV-tk) gene and the subsequent treatment with ganciclovir(GCV). Methods: The TK gene was transfected into human gastric carcinoma cell line MKN28 using HSV-TK that packed with PA317 cell, the sensitivity of MKN28TK cells to GCV was examined in vitro. Results: The retroviral-mediated HSV-TK gene can be transfected to MKN28 cells. The growth rate of MKN28 cells transfected with HSV-TK gene did not change. MKN28TK cells became significantly sensitive to GCV and had bystander effect. Conclusion: Transfection of gastric carcinoma with HSV-TK has higher transfection efficiency. MKN28TK cells are significantly sensitive to GCV.

Objective: To investigate the effect of protein ki nase C on signal transduction such as tyrosine phosphorylation, c-fos and c-ju n mRNA expression in antigen activated mast cells. Methods: RBL-2H3 cells either untreated or treated with phorbol 12-myristate 13 -acetate (PMA) were sensitized with anti-DNP IgE, and activated with DNP-BSA, histamine release and tyrosine phosphorylation were quantitatively measured by ELISA and flow cytometry, respectively. The effect of PKC on the ex pression of c-fos and c-jun in serum-deprived RBL-2H3 cells activated by DNP-BSA detected by ethidium staining of PCR-amplified cDNA, the amplified cDNA products were subjected to Southern blot hybridization using specific prob es to determine the veracity of amplification. Results: Tyr osine phosphorylation and histamine release were significantly reduced from (4.4 7±0.03)% to (2.79±0.07)% and (104.47±1.31) nmol/L to (60.75±1.38) nm ol/L, respectively, 45 min after DNP-BSA stimulation in sensitized cells pre treated with PMA for 48 h. Bands of the size predicted for the amplified cDNA we re obtained: 299 bp for c-fos, and 651 bp for c-jun, a decrease of 91% and 82% , respectively, for c-fos and c-jun mRNAs was observed in antigen stimulated c ells pretreated with PMA for 48 h. Conclusion: PKC plays an impo rtant role in modulating the tyrosine phosphorylation and histamine release resp onses and may upregulate the expression of c-fos and c-jun in antigen activate d mast cell.

Objective　To investigate the mechanism for that insulin facilitates increase of glucose uptake. Methods　The expression of myocardial GLUT4 polypeptide was determined by semiquantitative immunoblotting. The expression of GLUT4 mRNA was determined by semiquantitative Northern blotting. Results　After infusing insulin for 8　hours，the　expression　of　GLUT4 mRNA and GLUT4 polypeptide was significantly higher in canine myocardium than in　those　found normal ones. The glucose uptake was upregulated at　the　same　time．Conclusions　Our findings suggest that insulin facilitates the expression of GLUT4 mRNA and GLUT4 polypeptide in canine hearts. Enhanced GLUT4 expression is one of the important molecular mechanism by which myocardial cells enhance glucose uptake by insulin stimulation.

Objective To analyze the Ag NORs in T cells of different patients to find clues for the diagnosis and monitoring of cancer.Methods A KL 2 imaging system was used to analyze the areas of Ag NORs in T cells. T cells from peripheral blood were cultivated and stimulated, and silver staining was performed. The areas of Ag NORs were analyzed.Results A total of 1 046 nomal adults, 393 patients with non malignant diseases and 706 patients with malignant diseases were analyzed. Their average IS% values were (7.81?0.69)%, (7.85?0 72)% and 5.17?0.87 respectively. The IS% values above 6% were 99.8%, 94.7% and 10% to 35% respectively. No differences were observed in different kinds of cancers except breast cancer and rectum cancer whose IS% were (5.61?0.86)% and (6.10?0.92)% respectively, 75% and 26% of their IS% were less than 6% respectively. 83% to 90% of the IS% in the patients with other cancers were below 6%. In comparison with serum CEA, AFP and the like, Ag NORs in T cells were more powerful in diagnosis and monitoring of cancers.Conclusion The value of Ag NORs of T cells was significantly lower in patients with cancers and was statistically different from that in nomal adults and patients without malignant diseases. The results demonstrated that the analysis of Ag NORs might play an important role in the diagnosis, differenciation and monitoring of cancer.

Objective: To explore the molecular mechanism of HMC-1 cell apoptosis on exposure to tripterine. Methods: After the HMC-1 cells were incubated with tripterine, the expression of bcl-2, bax, bcl-X,c-myc and ICE were assayed by using immunohistochemical staining and RT-PCR. Results: The expression of bax,c-myc were up-regulated and bcl-2 down-regulated at protein level.The expression of bax,bcl-X L, especially bcl-2 were down-regulated, and ICE was up-regulated at mRNA level. Conclusion: These results suggest that apoptosis of HMC-1 cells induced by tripterine is regulated by different expression of apoptosis-related genes.