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Objective:To prepare the ocular thermosensitive gel of atropine sulfate and study its in vitro release. Methods: The gel formula was optimized by central composite design response surface methodology. The influences of the amounts of poloxamer 407 (P407) and poloxamer 188(P188) on gelling temperature before and after the dilution with simulated tear fluid were investigated. A membraneless dissolution model was used to determine the gel erosion and in vitro release. Results:The optimized gel formula was as follows:23% P407 and 5% P188. The deviations between the measured values and predicted values were all lower than 5%. The in vitro release experiment showed that the gel erosion and the drug release fitted zero-order kinetics equations with promising correlation, indicating a dissolution-controlled release mechanism. Conclusion:The optimization of the ocular thermosensitive gel of atropine sul-fate can be achieved by central composite design response surface methodology with good estimation. The thermosensitive gel with sus-tained drug release property meets the design requirements.
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Objective:To prepare the thermosensitive nasal gel of ephedrine hydrochloride and diphenhydramine hydrochloride and establish its quality control method. Methods:The amounts of P407, P188 and PEG 6000 were optimized by an orthogonal test with the gelling temperature as the index. An HPLC method was established to determine the contents of ephedrine hydrochloride and di-phenhydramine hydrochloride. Results:The optimum amount of P407, P188 and PEG 6 000 was 19%, 2% and 1%, respectively. The linear range of ephedrine hydrochloride was 1.600 0-2.400 0 mg·ml-1(r=0.999 6), and the average recovery was 99.76%with RSD of 1. 02%(n=9). The linear range of diphenhydramine hydrochloride was 0. 160 0-0. 240 0 mg·ml-1(r=0. 999 7), and the average recovery was 101. 27% with RSD of 1. 10%(n=9). Conclusion:The formula design and preparation technology of the gel are feasible. The HPLC method is suitable for the quality control of the preparation.
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Objective:To optimize the formula of Junduqing dispersible tablets .Methods:Using appearance , disintegration time and dispersing uniformity as the indices ,the types of filler, disintegrating agent and adhesive were optimized by single factor tests;the amount of filler and disintegrating agent was optimized by orthogonal experiments with the disintegration time as the index ; using ap-pearance and sticking as the indices , the single factor tests were also used to optimize the lubricant , and the adding method of disinte-grating agent was optimized using disintegration time as the index .Results: For 500 tablets, the amount of Junduqing extract was 61.25 g, microcrystalline cellulose (52.5 g) and dextrin (17.5 g) were used as the fillers, crospolyvinylpyrrolidone (8.75 g) and croscarmellose sodium (26.25 g) were applied as the disintegrating agents , 70% ethanol was the binder , magnesiumstearate (3 g) and talcum powder (2.25 g) were used as the lubricants , and stevioside (3.5 g) was the flavoring agent .Conclusion:The formula is reasonable and feasible ,which provides reference for the preparation of Junduqing dispersible tablets .
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This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.
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This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.
Subject(s)
Humans , Apoptosis , Cell Cycle , Cell Line, Tumor , HL-60 Cells , Hep G2 Cells , K562 Cells , Membrane Proteins , Metabolism , Pharmacology , Tumor Necrosis Factor-alpha , Metabolism , PharmacologyABSTRACT
Objective:To study the mechanisms of immune suppression mediated by Gr-1+CDllb+ myeloid derived suppressor cells (Gr-1~+CDllb~+MDSC)from tumor-bearing mice.Methods:Gr-1~+CDllb~+MDSC recruited into spleen and bone marrow of tumor-bearing mice were purified by Percoll,and suppression mediated by MDSC on T cell proliferation from spleen of naive mice was detected by flow cytometry with CSFE and FTTC-anti CD3 staining,and NO,ROS,IL-10 and TGF-β in the supematant of MDSC were detected by Griess and ELISA.Results:There were much more Gr-1~+CDllb~+MDSCs in spleen and bone marrow from tumor-bearing mouse than those of naive mouse,and suppression on T cell proliferation mediated by MDSC from tumor beating mouse was significantly increased,and there were much more NO,ROS,IL-10 and TGF-βin the supematant of these MDSC than that from naive mouse.Conclusion:MDSC from tumor-bearing mice secreted hish level of NO,ROS,IL-10 and TGF-βto induce immune suppression,and inhibite the proliferation of T cells.
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The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl-isomerase PIN1 gene, the mutation in exon 3 of beta-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of beta-catenin gene and differential expression of beta-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (chi2=32.63, P0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of beta-catenin, and accompanied by beta-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of beta-catenin gene in tissues with high PIN1 expression level (chi2=58.12, P<0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from beta-catenin gene mutation.
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The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl- isomerase PIN1 gene, the mutation in exon 3 of β-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of β-catenin gene and differential expression of β-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (x2 =32.63, P＜0.05), especially in cytoplasm and nucleus, while there was lower level of PIN1 expression in non-cancerous tissues; (2) 58.6% (17/29) HCC tissues showed β-catenin protein accumulation in cytoplasm and nucleus. 46.2% (6/13) HCC tissues indicated β-catenin protein accumulation with higher level of PIN1 expression, while 53.8% (7/13) HCC tissues indicated β-catenin protein accumulation with lower level or trace of PIN1 expression (x2 =0.00, P＞0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of β-catenin, and accompanied by β-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of β-catenin gene in tissues with high PIN1 expression level (x2=58.12, P＜0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from β- catenin gene mutation.
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In order to construct an expression vector carrying small hairpin (sh) RNA (shRNA) for toll-like receptor 4 mRNA and a reporter gene of enhanced green fluorescence protein (EGFP) and study the inhibition of cytokine release by RAW264.7 cell induced by lipopolysaccharide (LPS) stimulation through transfection and expression of shRNA targeting TLR4 gene via the RNAi mechanism, the reporter gene plasmid pEGFP-C1 (4.7 kb) and psiRNA-hHlneo (2979 bp) were used. The H1 promotor and double Bbs I restrict endoenzyme site were cloned from plasmid psiRNA-hH1neo and reconstructed them into plasmid pEGFP-C1 in the Mlu I restrict endoenzymic site, forming plasmid pEGFP-H1/siRNA, which contained Bbs site and reporter EGFP gene. Then an oligonuclear hairpin sequence targeting TLR4 gene was designed by internet tool and inserted into the plasmid pEGFP-H1/siRNA forming plasmid pEGFP-H1/TLR4-siRNA. After transfection of pEGFP-H1/TLR4-siRNA into RAW264.7 cells, tumor necrosis factor-alpha (TNF-alpha) release by the cells after stimulation by LPS was detected. The results showed that the constructed pEGFP-H1/TLR4-siRNA carrying hairpin RNA for TLR4 gene and reporter EGFP gene were proven to be right by restriction endonuclease analysis. The expression of EGFP gene was (50.37+/-8.23) % and after transfection of the plasmid pEGFP-H1/ TLR4-siRNA the level of TNF-alpha released by RAW264.7 cell was down regulated. It was concluded that shRNA targeting TLR4 gene could inhibit the TNF-alpha release by RAW264.7 cells evoked by LPS.
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In order to construct an expression vector carrying small hairpin (sh) RNA (shRNA) for toll-like receptor 4 mRNA and a reporter gene of enhanced green fluorescence protein (EGFP) and study the inhibition of cytokine release by RAW264.7 cell induced by lipopolysaccharide (LPS)stimulation through transfection and expression of shRNA targeting TLR4 gene via the RNAi mechanism, the reporter gene plasmid pEGFP-C1 (4.7 kb) and psiRNA-hHlneo (2979 bp) were used. The H1 promotor and double Bbs Ⅰ restrict endoenzyme site were cloned from plasmid psiRNA-hHlneo and reconstructed them into plasmid pEGFP-C1 in the Mlu Ⅰ restrict endoenzymic site, forming plasmid pEGFP-H1/siRNA, which contained Bbs site and reporter EGFP gene. Then an oligonuclear hairpin sequence targeting TLR4 gene was designed by internet tool and inserted into the plasmid pEGFP-H 1/siRNA forming plasmid pEGFP-H 1/TLR4-siRNA. After transfection of pEGFP-H1/TLR4-siRNA into RAW264.7 cells, tumor necrosis factor-alpha (TNF-α) release by the cells after stimulation by LPS was detected. The results showed that the constructed pEGFP-H1/TLR4-siRNA carrying hairpin RNA for TLR4 gene and reporter EGFP gene were proven to be right by restriction endonuclease analysis. The expression of EGFP gene was (50.37±8.23) %and after transfection of the plasmid pEGFP-H1/ TLR4-siRNA the level of TNF-α released by RAW264.7 cell was down regulated. It was concluded that shRNA targeting TLR4 gene could inhibit the TNF-α release by RAW264.7 cells evoked by LPS.
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To evaluate the specific inhibition of antisense u-PAR on the u-PAR expressions in highly invasive cell subclones and to determine its blocking function in the invasion by those cells, a cDNA fragment of u-PAR obtained by RT-PCR was inserted into a plasmid vector named pcDNA3 in antisense orientation. Then the antisense u-PAR recombinant was transfected into highly invasive cell subclones. The u-PAR expression in neo-resistant cells was examined by RT-PCR and immunohistochemical assay. Compared to the control cells, the content of mRNA and protein of u-PAR in transfected cells decreased sharply, and the rate of inhibition was 53% and 73%, respectively, indicating that an antisense u-PAR might have played a specific inhibitory role in its expression in the cells, which may provide a good cell model for making further investigation of the inhibitory effects of the antisense u-PAR on invasion in highly invasive cell subclones of human prostate carcinoma.
Subject(s)
Humans , Male , Cell Line, Tumor , Cloning, Molecular , Neoplasm Invasiveness , Plasmids , Prostatic Neoplasms , Metabolism , Pathology , RNA, Antisense , Genetics , Receptors, Cell Surface , Genetics , Receptors, Urokinase Plasminogen Activator , Reverse Transcriptase Polymerase Chain Reaction , Transfection , Urokinase-Type Plasminogen Activator , Genetics , MetabolismABSTRACT
To observe the inhibitory effects of an antisense u-PAR vector on invasion of highly invasive PC-3M cell subclones, the effects of the antisense u-PAR on activity of MMP-9 in those highly invasive cell subclones were detected by a quantitative RT-PCR and zymography. The monolayer invasion assay and colony formation assay in soft agar were used. And tumorigenesis rate and invasions by the cell subclones with or without the antisense u-PAR were observed in nude mice. It was found that in vitro growth of highly invasive PC-3M cell subclones transfected with the antisense u-PAR was declined, and the ability of anchorage-independent growth of those cell subclones was found decreased sharply, with the inhibiting rate becoming 79% and 60%, respectively. Although the antisense u-PAR didn't change MMP-9 gene transcription, they could inhibit the activation of MMP-9 of highly invasive PC-3M cell subclones. Moreover, the tumorigenesis rate of the cell subclones with the antisense u-PAR decreased and the growth of a neoplasm also slowed down. The t tests showed the difference between experimental and control groups was statistically significant (P < 0.01). The antisense u-PAR vector could not only inhibit the invasion ability of highly invasive PC-3M cell subclones in vitro but also restrain the growth of those cell subclones in vivo.
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Animals , Humans , Male , Mice , Antisense Elements (Genetics) , Genetics , Pharmacology , Cell Division , Cloning, Molecular , Matrix Metalloproteinase 9 , Genetics , Metabolism , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness , Prostatic Neoplasms , Metabolism , Pathology , RNA, Antisense , Receptors, Cell Surface , Genetics , Metabolism , Receptors, Urokinase Plasminogen Activator , Reverse Transcriptase Polymerase Chain Reaction , Transfection , Tumor Cells, CulturedABSTRACT
<p><b>OBJECTIVE</b>To investigate the regulation of leptin on insulin secretion and expression of ATP-sensitive potassium channel subunit sulfonulurea receptor 1 (SUR1) mRNA, and to determine whether the effects of leptin are mediated through known intracellular signaling transduction.</p><p><b>METHODS</b>Pancreatic islets were isolated by the collagenase method from male SD rats. The purified islets were incubated with different concentrations of leptin for 2 h in the presence of different concentrations of glucose. Insulin release was measured using radioimmunoassay. Expression of SUR1 mRNA was detected by RT-PCR.</p><p><b>RESULTS</b>In the presence of leptin 2 nmol/L, insulin release was significantly inhibited at either 11.1 or 16.7 mmol/L glucose concentration (both P < 0.05), but insulin release was not altered at glucose of 5.6 mmol/L physiological concentration. The dose-response experiment showed that the maximal effect of leptin on insulin secretion achieved at 2 nmol/L. Exposure of islets to 2 nmol/L leptin induced a significant increase of SUR1 transcription levels by 71% (P < 0.01) at 11.1 mmol/L glucose and by 56% (P < 0.05) at 16.7 mmol/L glucose concentration. Selective phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin significantly prevented the leptin effect on insulin secretion and SUR1 mRNA expression.</p><p><b>CONCLUSIONS</b>Regulatory effects of leptin on insulin secretion could be biphasic at different concentrations of glucose and leptin. The stimulatory regulation of SUR1 transcription levels may be mediated through activation of PI 3-kinase pathway, which may be a possible mechanism of leptin in regulating insulin secretion.</p>
Subject(s)
Animals , Male , Rats , Butadienes , Pharmacology , Cells, Cultured , Dose-Response Relationship, Drug , Insulin , Bodily Secretions , Islets of Langerhans , Metabolism , Leptin , Pharmacology , Nitriles , Pharmacology , Phosphatidylinositol 3-Kinases , Physiology , Potassium Channels, Inwardly Rectifying , Genetics , RNA, Messenger , Rats, Sprague-DawleyABSTRACT
To compare the anti-tumor effects of transmembrane TNF-alpha (TM-TNF) and secreted TNF-alpha (S-TNF) in vivo, mouse fibroblasts NIH3T3 were transfected separately with three types of retrovirus containing wild type TNF-alpha (Wt-TNF), TM-TNF mutant (TM-TNFm), S-TNF mutant (S-TNFm). Southern blot, RT-PCR, FACS and bioassay were used to investigate TNF-alpha gene integration, expression and its biological activity. It was found that both fixed cells and supernatant of NIH3T3/Wt-TNF, the fixed cells of NIH3T3/TM-TNFm and the supernatant of NIH3T3/S-TNFm could express high level of TNF-alpha or its mutants and effectively kill H22 in vitro. The transfected NIH3T3 were separately injected into the mice at the sites of H22 tumor cell inoculation according to a ratio of 5:1 or 1:1 (effector/target cells, E/T) after the third day of H22 challenge, respectively. At the E/T = 5:1, the NIH3T3/TM-TNFm induced the highest tumor regression, while NIH3T3/S-TNFm exerted the strongest tumor depressing effect at the E/T = 1:1 in vivo. No obvious side effects were noted throughout the course of treatment. The results suggest that both TM-TNF and S-TNF could cause tumor regression. The anti-tumor effect of TM-TNF would be more powerful and safe than that of S-TNF at the proper E/T ratio.
Subject(s)
Animals , Mice , 3T3 Cells , Adenoviruses, Human , Genetics , Cytotoxicity, Immunologic , Allergy and Immunology , Fibroblasts , Cell Biology , Allergy and Immunology , Liver Neoplasms, Experimental , Allergy and Immunology , Pathology , Membrane Proteins , Bodily Secretions , Mutation , Transfection , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha , Genetics , Allergy and Immunology , Bodily SecretionsABSTRACT
<p><b>OBJECTIVE</b>To observe the dynamic expression of transmembrane (TM)-tumor necrosis factor (TNF)-alpha and secreted (S)-TNF-alpha in the development of endotoxic shock and explore the actions and mechanism of TM-TNF-alpha in liver of the rat with endotoxic shock.</p><p><b>METHODS</b>Endotoxic shock in rats was induced by intravenous injection of dead gram negative bacteria E. Coli; the kinetics of TM-TNF-alpha on peritoneal macrophages and S-TNF-alpha in serum of these rats were determined. Pretreatment with TNF alpha converting enzyme antisense oligonucleotide (5 mg/kg) 30 minutes before rats were administrated dead bacteria inhibited enzymatic cleavage of TM-TNF-alpha into S-TNF-alpha. Six hours after bacteria injection, TM-TNF-alpha and S-TNF-alpha were also detected respectively. The pathological injury in the livers of rats with endotoxin shock was examined, and artery pressure was constantly measured.</p><p><b>RESULTS</b>The kinetics of TM-TNF-alpha expression in the development of endotoxic shock was different from that of S-TNF-alpha expression in serum. The expression of TM-TNF-alpha began to increase on the surface of peritoneal macrophages and liver within 30 min, after bacteria challenge and peaking within a period of 4.5 hours followed by a gradual decrease to a relatively high level which was maintained for at least 24 hours. The TACE antisense oligonucleotide pretreated rats showed remarkable increase in TM-TNF-alpha expression by peritoneal macrophages and liver (P < 0.001), and their arterial blood pressure were maintained within normal levels and there were no detectable pathological changes in their livers.</p><p><b>CONCLUSIONS</b>These findings suggested that TM-TNF-alpha may be a potent endogenous regulator involved in anti-inflammatory responses to maintain normal arterial pressure and protect liver tissue from pathological injury in during endotoxin shock. This study confirmed the important role of TNF-alpha in endotoxic shock which is not only of important theoretical significance, but also of practical interest in providing experimental basis for clinical treatment of endotoxin shock.</p>
Subject(s)
Animals , Rats , Chemical and Drug Induced Liver Injury , Disease Models, Animal , Endotoxins , Toxicity , Liver Diseases , Metabolism , Membrane Proteins , Bodily Secretions , Oligonucleotides, Antisense , Pharmacology , Rats, Wistar , Shock, Septic , Metabolism , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
AIM: To compare the tumorigenecity of H22 cells transfected with TNF-? gene and its mutants (secreted TNF-? mutant, S-TNFm, transmembrane TNF-? mutant, TM-TNFm and wild type of TNF-?, Wt-TNF) in vivo . METHODS: Three kinds of mouse liver cancer cell line H22 expressing TNF-? and its two mutants were mixed with untransfected H22 at different effector/target ratio separately. The growth of tumor was examined after injection of 2.5?10 5 (100 ?L) mixed H22 tumor cells into mice. The lymphocyte infiltration in the site of tumor and the expression of Fas on tumor cells were detected by immunohistochemistry. RESULTS: The tumorigenecity of H22 cells transfected with TNF-? gene and its mutants was significantly weakened ( P
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Objective:To construct and express the recombinant of human soluble TNF receptor I by E coli Methods:The cDNA coded for extracellular region of human TNFRI was amplified by RT PCR and inserted into a expression vector, pET 28a Then, the recombinant sTNFRI/pET 28a was transfected and expressed in E coli Results:After the stimulation with IPTG, sTNFRI fusion protein was effectively produced in E coli BL 21 transfected with the exogenous gene A unique band was found at 27 kD by SDS PAGE and its expression was about 31% of total protein in E coli The purified sTNFRI fusion protein was shown to be able to suppress the cytotoxicity of TNF on L929 cells and found by indirect immune fluorescence to inhibit specifically the binding of TNF with TNFR on target cells Conclusion:The recombinant human soluble TNFR I have been obtained by using the genetic engineering technology
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Objective:To study the effects of secreted stem cell factor(S SCF) and transmembrane SCF(TM SCF) on the biological functions of mast cells,including proliferation,release of histamine and secretion of cytokine to explore the biological characteristics of the two forms of SCF.Methods:MTT,fluorescence technique,in situ hybridization were used to detect the several biological functions of mast cells,such as proliferation,release of histamine and the transcription of TNF ? mRNA.Results:It was shown that S SCF could significantly promote the proliferation of mast cells,release of histamine and TNF ? mRNA transcription,while TM SCF had only a weaker positive effect on histamine relase and no any effect on proliferation and TNF ? mRNA transcription.Conclusion:The biological actions of TM SCF on mast cells are different from that of S SCF.
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Objective:To study the effects of PTK on the respiratory burst and release of NO by neutrophils in response to transmembrane TNF-?( TM-TNF-?) and secreted TNF-? ( S-TNF-?). Methods: The effects of PTK inhibitor on the functions of neutrophils caused by both forms of TNF-? was detected with the chemiluminescence and the nitrate reductase assay.The tyrosine phosphorylation of neutrophils induced by the two forms of TNF-? was compared by Western blotting analysis. Results: The PTK inhibitor genistein (50 ?mol/L) was found not only to depress the respiratory burst of neutrophils induced by S-TNF-?( P
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Abstract Objective:The effects of both TM-TGFa and S-TGFa on proliferation were compared to explore the biological characteristics of the two forms of TGF-a.Methods:The proliferation of mouse fibroblast cell line NIH3T3 and its expression of PCNA were determined by the methods of MTT and histochemical technique. The level of IL-6 mRNA in HaCaT cell line was tested by in situ hybridization. Results:The two forms of TGFa were able to promote cell proliferation,PCNA expression and IL-6 mRNA transcription.The effects of S-TGFa had been shown stronger than that of TM-TGFa( P