Objective Investigation of the effect of integrin on fibroblasts from scleroderma in the production of procollagen. Methods Phosphorothioate modified antisense oligonucleotides were used to interfere with the expression of integrin ? 5 or ? 1 subunit on scleroderma fibroblasts respectively, then the changes of procollagen mRNA were detected by RT PCR. Results The expression of integrin ? 5 or ? 1 subunit could be specifically inhibited by their antisense oligonucleotides respectively. Decreased expression of fibroblast integrin ? 5 or ? 1 subunit significantly lowered mRNA of procollagen ? 1(Ⅰ) and ? 1(Ⅲ). Conclusion Overproduction of procollagen may be inhibited in the level of transcription by lowering the expression of integrin on fibroblasts in scleroderma.

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.

Hepatitis C virus (HCV) is a major causative agent for sporadic and post-transfusion hepatitis, which frequently progresses into chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. In order to clarify the genomic variations and the structural characteristics of Chinese HCV isolate, we extracted HCV genomic RNA from plasma specimens of Chinese hepatitis C (HC) patients, reverse-transcribed to HCV cDNA with random primers and constructed successfully an HCV cDNA R gt11 library of Chinese type. Two positive clones (Q349 and Q653) were selected by immunoscreening from the library and subcloned into plasmid vector pUC18. Sequence analyses indicated that Q349 was derived from core region (positions 554-902) of HCV genome while Q653 was from NS3 region (positions 4175-4827) corresponding with the prototype HCV nucleotide sequence. The homologies of Q349 and Q653 with the equivalent sequences of HCV prototype were 86.8% and 80.2% at the nucleotide level, and 97.3% and 93.1% at the amino acid level, respectively. It was found that Chinese HCV clones had higher homologies with Japanese HCV isolates, and should belong to HCV group II. Specificity test proved that the encoded peptides of the 2 Chinese HCV cDNA clones reacted specifically with sera from HC patients and had no reaction with sera from healthy individuals. More importantly, clone Q653 showed higher positive reaction rates with Chinese HC sera (95.8%) than those with Japanese ones (85.7%), which strongly suggests that the sequences from Chinese HCV genome (especially from NS regions) would be more suitable for primer designing or peptide synthesis for the use in the detection of HCV infection among Chinese people.

AIM: To observe the expression of undulin(Un) in liver tissue and to clarify the diagnostic significance of serum Un in experimental rat liver fibrosis. METHODS: Rat liver fibrosis was induced by carbon tetrachloride. The expression of Un in liver tissue was detected by immunohistochemical method. Serum Un levels was measured by enzyme immunoassay.RESULTS: Expression of Un increased in fibrotic liver than normal liver, and it was mainly distributed in portal tract stroma, central veins and fibrotic septa in fibrotic liver. Also, the level of serum Un was significantly higher in fibrotic liver than normal liver.CONCLUSION: These data suggest that Un should be a component of the hepatic extracellar matrix, and its expression could be increased greatly in fibrotic rat liver. Serum Un levels may be used as an indicator in liver fibrosis diagnosis.

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.

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 effects of TGF-?1 on promoter activity of human transforming growth factor-beta1(TGF-?1) gene.Methods:The fragments with different length of the 5′-end sequence of the human TGF-?1 gene between -1 328 bp to +812 bp were obtained from a healthy male person genomic DNA, and the sequences were fused to chloramphenicol acetyltransferase reporter gene in the pCAT-enhancer plasmid to construct five chimeric recombinant plasmids. These recombinant plasmids were transiently transfected into rat hepatic setellate cell line(nFSC) by FuGENE6 transfection methods. The cells transfected with chimeric recombinant plasmids were cultured on media in the absence or presence of TGF-?1(2 5 ng/ml),CAT activities in transfected cells were tested and compared.Results:The different concentration of TGF-?1 can prevent the proliferation of nFSC, and these effects are dose-dependent; The presence TGF-?1 can stimulate the CAT activity of cells transfected with phTGF0 585?phTGF1 120?phTGF1 423?phTGF1 680?phTGF2 140 up to 2～5 times higher then cells cultured with absence of TGF-?1.Conclusion:The TGF-?1 can stimulate CAT activities in nFSC transfected with phTGF0 585?phTGF1 120?phTGF1 423?phTGF1 680?phTGF2 140. These results suggest that TGF-?1 have an autoregulation effect on TGF-?1 gene.

38 cases of Benign Monoclonal Gammopathy (BMG) representing 17.9% were screened from 212 cases of M-proteinemia by laboratory examinations. Among the 38 cases, 17 cases had increase of IgG, another 17 cases of IgA and 4 cases of IgM. Their average M-proteins were 1.42, 0.88 and 1.84 g/dl respectively.The salient points of differential of benign or malignant M-proteinemia were also discussed. The recommended sequential laboratory diagnostic procedure of BMG were as follows: tatol serum protein determination, plasma protein zone electrophresis, urine protein analysis, immunoglobulin quantitation and immunoelectrophresis or selective electrophresis.

Experiment Ⅰ. Twenty-eight rats were randomly assigned into Near-Infrared Information Radiation (NIIR) group and control group. Two weeks later each rat was innoculated intraperitoneally with Salmonella typhosa H antigen (HAg) and cyclophosphamide (CY). Peripheral lymphocyte counts in the NIIR group were significantly higher than those in the control group on the fifth day after administration of CY. Experiment Ⅱ, Fifty-four rats were randomly divided into NIIR group given CY and H Ag intraperitoneally, CY and H Ag group and H Ag group for treatment. By the end of the fourth week, the survival rate and serum IgG level in the NIIR group were significantly higher than those in the CY and HAg group. By the end of 2nd week, the titer of the anti-H antibody of the HAg group and NIIR group was significantly higher than that of the CY and HAg group. Experiment Ⅲ. Thirty rats were randomly allocated to NIIR group and control group. Spleen cells were taken and cultured with Con A for 24h to induce IL-2 and the activity of IL-2 in the NIIR group was significantly higher than that in the control group. The NK activity in NIIR group was higher but not significant and ADCC in the NIIR group was significantly higher than that in the control group. The results suggest that NIIR is capable of enhancing immunoreaction in immunosuppressive bodies by promoting the function recovery of T helper cells, therefore NIIR is effective to regulate the immunological function on chronic active hepatitis.

Four cell lines (C6, H6, B6, 2H6) of hybrids secreting monoclonal antibodies (McAb) against erythrocytic stage of Plasmodium falciparum Fcc7801/HN were established by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immuniaed with the preparation of the same malaria parasite and cloning three times. Of these four lines, C6 and B6 showed significant growth-inhibition of P. falciparum in vitro. Both C6 and B6 McAb recognized 71 kD antigen of P. falciparum by Western Blotting. The results indicate that the 71 kD antigen is of immune protection The four McAb showed cross immunofluorescence reactivity with P. falcipanum Fcc-1/HN, Fcc7802/HN, Fcc8703/JS and P. berghei. P. cynomolgi. It indicates that there are common epitopes among these malaria parasites, and that it is important in immunologic research and serodiagnosis of malaria