A Comparative Study on the Response of Dermal Fibroblasts from Normal and Hypertrophic Scar-ring Skin to Cytokines,Extracellular Matrix and Collagen Promotor / 中华皮肤科杂志

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.

Clinical implications of increased expression of CD40L in patients with acute coronary syndromes / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate clinical implications of expression of CD40L in monocytes and changes in serum soluble CD40L in patients with acute coronary syndromes (ACS).</p><p><b>METHODS</b>Sixteen 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.</p><p><b>RESULTS</b>Expression 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.</p><p><b>CONCLUSION</b>Enhanced level of serum sCD40L may be a reliable prognostic indicator for ACS and may represent a marker of coronary disease activity.</p>

Dynamic expression of tenascin in rat liver during liver fibrogenesis induced by CCl(4) / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression of tenascin in normal and fibrotic rat liver.</p><p><b>METHODS</b>Liver 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.</p><p><b>RESULTS</b>In 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.</p><p><b>CONCLUSIONS</b>Tenascin is a component of the extracellular matrix of liver tissue. Plays a role in early matrix organization during liver fibrogenesis.</p>

Clone of M2 autoantigens from human sources and evaluation of measurement of M2 autoantibodies in diagnosis of primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the value of measurement of M2 autoantibodies in diagnosis of primary biliary cirrhosis (PBS).</p><p><b>METHODS</b>M2 autoantibodies were detected by ELISA combined with western-blot.</p><p><b>RESULTS</b>Thirty-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.</p><p><b>CONCLUSIONS</b>M2 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</p>

Effect of tripterine on expression of bcl-2 family, c-myc and ICE gene in HMC-1 cells / 第二军医大学学报

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.

Effect of protein kinase C on signal transduction in antigen activated mast cells / 第二军医大学学报

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.

Transfecting and killing effect of herpes simplex virus thymidine kinase (HSV-TK) gene on human gastric carcinoma in vitro / 第二军医大学学报

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.

Insulin enhances glucose transporter 1 gene expression of low-flow ischemic myocardium / 中国病理生理杂志

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.

Significance of nucleolar organizer regions associated proteins of T lymphocytes in diagnosis and mornitoring of cancer / 中华检验医学杂志

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.

Insulin facilitates glucose transporter 4 gene expression in canine heart in vivo / 免疫学杂志

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.

The effect of La protein on replications of HBV in cultured cell / 中国免疫学杂志

Objective:To prove whether La protein could effect the replications of the HBV in cultured cells.Methods:Three specific SiRNAs for human La protein was obtained by transcription in vitro. After transfection with the SiRNAs into HepG2.2.15 cell, the levels of La protein mRNA and HBV DNA were detected by real-time PCR.Results:The HBV DNA secreted by the cell transfected with the SiRNAs was reduced with reduction of the mRNA of the La protein.Conclusion:The La protein can protect the replications of the HBV in cultured cells.

Effect of Integrin Expression on Procollagen Synthesis of Fibroblasts from Scleroderma / 中华皮肤科杂志

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.

Expression of undulin in experimental rat liver fibrosis / 中国病理生理杂志

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.

Effect of TGF-?1 on TGF-?1 gene promoter activity in mouse hepatic satellite cells / 中国免疫学杂志

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.

Extracellular Matrix in Liver: Experimental Study and Clinical Application / 第二军医大学学报

Laminin(LN) and hyaluronie acid (HA) are extracellular matrix in liver. The effects of LN and HA on human fetal liver cells proliferation and collagen synthesis were observed and serum levels of LN and HA were assayed m patients with liver diseases by an enzyme immunoassay method. Both LN and HA had a significant effect on the inhibition of cell proliferation and collagen production. However, serum levels of LN and HA were significantly increased in liver cirrhosis comparing with healthy controls and with patients with non-liver diseases. The results suggest that LN and HA may play a role in the feed-back inhibition mechanism in liver fibrogencsis, and assay of LN and HA in serum may be useful to the diagnosis of liver fibrosis.

Immunoscreening and Identification of Chinese HCV Genomic cDNA ? gt11 Library / 第二军医大学学报

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.

DETECTION OF IL-4-PRODUCING CELLS IN PERIPHERAL BLOOD OF MULTIPLE MYELOMA PATIENTS AND ITS IMPLICATIONS / 解放军医学杂志

Using the APAAP method, we detected IL 4 producing cells in the peripheral blood of 9 patients with multiple myeloma and 11 normal subjects and lound that positive cell in MM (7.6%?2.1%) were significantly less as compared to normal subjects (15.36% ?4.1%), (P

T Subsets and Antitumor Activity of Lymphocytes Infiltrating Hunan Primary Brain Gliomas / 第二军医大学学报

Glioma-infiltrating lymphocytes (GIL) were isolated from 9 surgical biopsy specimens of primary brain gliomas using mechanical and enzymatic digestion and discontinuous density gtadient centrifugation. During cultured in the presence of interieukin-2 (IL-2) for a period of four weeks, GIL were expanded 48.4-fold on the averags, even up to 118-fold. GIL activated by IL-2 had specific cytorytic activity against autologous glioma cells. Analysis of T subsets of GIL freshly isolated showed that CD3+ cells were 71.0?11.9%, CD4+ cells 34.2?6.1% and CD8+ cells 37.0?7.6%. Ability of activated GIL to secrete ?-interferon (?-IFN) was significantly higher than that of freshly isolated GIL and autologous peripheral blood lymphocytes (PBL). The results suggest that GIL have many advantages for an adoptive immunotherapy of patients with brain gliomas and is a new type of antitumor immune effector.

LABORATORY DIAGNOSIS OF BENIGN MONOCLONAL GAMMOPATHY: ANALYSIS OF 38 CASES / 解放军医学杂志

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.

High dose of intravenous immunoglbulin in treatment of presensitized patients awaiting kidney transplantation patients / 第二军医大学学报

Objective: To investigate the effect of intravenous immunoglobulin in highly sensitized patients awaiting organ transplantation. Methods: IVIG was used to reduce donor specific anti HLA alloantibodies in vitro and in vivo . Fifteen patients received IVIG′s suppressive experiment in vitro by random panel lymphocytotoxicity test. The serum of patients were divided into 2 groups: one was diluted with equal volume of IVIG and the other was diluted with PBS solution, and then reacted with lymphocytes from healthy donors randomly. Of them 5 patients received the treatment by IVIG. Four patients were administrated with 0.5 g/kg. Period of treatment was 4 weeks. One patient received 8 weeks infusion in same dose, 2 patients resulted in PRA drop to 10% had received kidney and pancreas kidney transplantation. Results: The percentage of RPLT in experimental group was lower than that in control group. After large dose of infusion of IVIG, the patients showed a reduction in absolute PRA of 2% 51% (mean decrease: 23%). Two patients had undergone subsequent transplantation and no serious rejection occurred. Conclusion: Treatment with IVIG is a valuable tool for the transplantation of immunized patients. The effect of IVIG is dose dependent and can be achieved in 3 weeks.