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Chinese Medical Sciences Journal 1996年第1期11卷 25-27页

作者：郭子健朱元珏刘秉慈朱亚玲赵文理陈勇 Department of Internal Medicine PUMC Hospital Beijing.

Progressive inflammation and fibrosis are the central processes in the pathogenesis of pulmonary fibrosis. It is believed that macrophages in areas of chronically inflamed lung play a key role in fibrotic response. Therefore, we investigated the effects of alveolar macrophage (AmΦ) conditioned media from interstitial lung disease (ILD) patients on lung fibroblast proliferation and procollagen mRNA expression. After stimulating with AmΦ conditioned media from ILD patients, the fibroblast proliferation increased 71. 4 % compared with the control, but for media from bronchial carcinoma (BC) patients, it just increased 14. 3%. There is a significant difference between the two groups (Pprocollagen a1 (Ⅰ) mRNA in fibroblasts stimulated with AmΦ conditioned media from ILD patients was increased 21. 3 %, and a1 ( Ⅲ) was 37. 2% higher than control (P<0.05). It increased 6. 8% and 12. 8% for media from BC patients respectively, but there was no difference when compared to the control. We considered that AmΦ from ILD patients might be in an activated state and could release some growth factors to stimulate fibroblast proliferation and promote collagen DNA expression.

关键词： interstitial lung disease alveolar macrophage procollagen

In situ localization of procollagen gene expression on cryosections of undecalcified fracture callus

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Chinese Journal of Traumatology 2000年第1期3卷 26-29页

作者：史炜镔杜宁符诗聪张昊程枫柴本甫 Department of Traumatology Ruijin Hospital Shanghai 200025 China.

To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. Results: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro α1(Ⅲ) in fibroblasts and some chondrocyte like cells were dominant; and at the end of second week high expression of type Ⅱ procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type Ⅰ procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous ***: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional technique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.

关键词： Fractures Callus In situ hybridization procollagen Gene expression

Effect of cytokines on promoter activity in mouse α2(Ⅰ) procollagen gene

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Journal of Medical Colleges of PLA(China) 1999年第1期14卷 12-16页

作者：高春芳王皓伍严安孔宪涛

Objective:To investigate the effect of TGFβ, IFNs and TNFα on promoter activities in mouse α2(I )procollagen gene. Methods: Recombinant constructions PCOL 0. 4, pCOL0. 8, pAZ1009 containing CAT reportergene fused respectively with 400, 800 and 2 000 hp length of the 5'--flanking region in mouse α2(Ⅰ) procollagengene were used for transfection. Promoter activities were determined by CAT reporter gene assay(CAT--ELISA) intransfected cells treated with different cytokines. Results: TGFβ(2 ng/ml ) increased the activities of -- 2kb-- +54hp, -- 780~ + 54hp and -- 348~ +54 hp fragments as promoter respectively, with the increase of activity of 2kb -- + 54hp being most evident. TNFα (100 u/ml ) decreased the activity of -- 2kb ~ + 54hp sequence aspromoter. A slight increase, but not decrease of CAT activities was found in cells treated with IFNα(1000 u/ml)or IFNγ(1 000 u/ml) when pAZ1009 was trans feeted. But CAT expression was decreased more greatly if IFNα orIFNγ was used in combination with TNFα compared with TNFα treatment alone. The same change was seen ifPCOLO. 4 or PCOLO. 8 was used for transfection. Conclusion: Positive TGFβ--responsive elements exist within Zkbflanking region in mouse α2 (Ⅰ ) procollagen gene mediating the fibrogenic effect, while some negative elementsmediate the antifibrotic effect of TNFα. The target antifibrotic effect of IFNs might lie outside this 2kb-- +54bpregion or posttranscriptionally.

关键词： procollagen promoter cytokines

Effect of mitomycin on normal dermal fibroblast and HaCat cell:an in vitro study

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2012年第12期13卷 997-1005页

作者： Yao-wen WANG Ji-hao REN Kun XIA Shu-hui WANG Tuan-fang YIN Ding-hua XIE Li-hua LI Otolaryngology-Head and Neck Surgery Department Ningbo First HospitalNingbo 315000China Otolaryngology-Head and Neck Surgery Department the Seeond Xiangya HospitalCentral South UniversityChangsha 410001China State Key Laboratory of Medical Genetics Central South UniversityChangsha 410001China

Objective:To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (HaCat cell),particularly the effect of mitomycin on intracellular messenger RNA (mRNA) synthesis of collagen and growth factors of ***:The normal dermal fibroblast and HaCat cell were cultured in *** cultures were exposed to 0.4 and 0.04 mg/ml of mitomycin solution,and serum-free culture medium was used as *** cellular morphology change,growth characteristics,cell proliferation,and apoptosis were observed at different *** the fibroblasts,the mRNA expression changes of transforming growth factor (TGF)-β1,basic fibroblast growth factor (bFGF),procollagen Ⅰ,and Ⅲ were detected by reverse transcription polymerase chain reaction (RT-PCR).Results:The cultured normal human skin fibroblast and HaCat cell grew exponentially.A 5-min exposure to mitomycin at either 0.4 or 0.04 mg/ml caused marked dose-dependent cell proliferation inhibition on both fibroblasts and HaCat *** morphology changed,cell density decreased,and the growth curves were without an exponential *** fibroblast proliferated on the 5th day after the 5-min exposure of mitomycin at 0.04 mg/***,5-min application of mitomycin at either 0.04 or 0.4 mg/ml induced fibroblast apoptosis but not *** apoptosis rate of the fibroblast increased with a higher concentration of mytomycin (pprocollagen Ⅰ and Ⅲ,and a marked increase in the mRNA production of ***:Mitomycin can inhibit fibroblast proliferation,induce fibroblast apoptosis,and regulate intracellular protein expression on mRNA *** additon,mitomycin can inhibit HaCat cell proliferation,so epithelial cell needs more protecting to avoid mitomycin's side effect when it is applied clinically.

关键词： Mitomycin Fibroblast HaCat cell Apoptosis TGF-β1 bFGF procollagen

Effects of Fuzheng Huayu 319 recipe on liver fibrosis in chronic hepatitis B

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World Journal of Gastroenterology 1998年第4期4卷 76-81页

作者： LIU Ping LIU Cheng XU Lie-Ming Hu Yi-Yang XUE Hui-Ming LIU Cheng-Hai ZHANG Zhi-Qing Institute of Liver Diseases Shanghai Acad. Traditional C. Shanghai 200032 China Third Huaiyin City Hospital Jiangsu Province China Shanghai University of TCM China Department of Hepatology China Institute of Liver Diseases Shanghai Acad. Traditional C. Shanghai 200032 530 # LingLing Rd China

AIM To investigate clinic effects of Fuzheng Huayu 319 recipe (319 recipe) on liver fibrosis in chronic hepatitis *** Ninety-five patients with chronic hepatitis B were divide into the treated (63 cases) and control (32 cases) group, and orally administrated with 0.5g 319 capsule or 0.5g Dahuang Zhachong pill tid for 3 months, respectively. The liver functions and serological fibrotic markers were observed before and after treatment, 12 cases in the treated group were examined with liver *** Three hundreds nineteen recipe could remarkably decreased serum ALT level and total bilirubin and significantly improve serum albumin and A/G ratio. Its effects were better than Dahuang Zhachong pill. Before treatment, patients' serum monamine oxidase activities, tissue inhibitor of metalloproteinase (TIMP)-1, procollagen type Ⅲ and laminin were all higher than those of health peoples. These levels decreased remarkably after treatment, and urine hydroxyproline level increased significantly (P＜0.001-0.05). Compared with the control, the improvement in treated group was better than that in the control except TIMP-1. According to the scoring system for staging of chronic hepatitis, the fibrotic extents of 7 cases among 12 cases examined by liver biopsy decreased remarkably (1 case decreased by 3 scores, 5 by 2 scores, 1 by 1 score).CONCLUSION Fuzheng Huayu 319 recipe had good therapeutic effects on chronic hepatitis B, it could reverse the development of liver fibrosis to some extent. In general its effects were better than that of Dahuang Zhachong pill.

关键词： liver cirrhosis/therapy hepatitis B Fuzheng Huayu procollagen hyaluronic acid metalloproteinases hydroxyproline

Dynamic changes of typeⅠ,Ⅲand N collagen synthesis and distribution of collagen-producing cells in carbon tetrachloride-induced rat liver fibrosis

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World Journal of Gastroenterology 1999年第5期5卷 397-403页

作者： DU Wei-Dong ZHANG Yue-E ZHAl Wei-Rong and ZHOU Xiao-Mei(Department of Pathology, Shanghai Medical University, Shanghai200032, China)(National Laboratory for Oncogenes and Related Genes, ShanghaiCencer Institute)See invited commentary on page 388 Department of Pathology Shanghai Medical University Shanghai 200032 China National Laboratory for Oncogenes and Related Genes Shanghai Cancer Institute China Department of Molecular Biology Max-Plank-Institute for Biochemistry Germany Department of Pathology Shanghai Medical University Shanghai 200032 138 Yixueyuan Road China

AIM To find out the relationship between the gene transcription of different types ofprocollagen and the deposition of the relevant collagens in the liver tissue and to confirm the types of collagen producing cells in *** Dynamic changes of the expression of α1(Ⅰ ), α1 (Ⅲ ) and α1 (Ⅳ) procollagen mRNAand relevant collagens and the distribution ofcollagen producing cells during liver fibrogenesis of rat induced by CCl4 (20 weeks)were investigated with Northern blot analysis,in situ hybridization and *** The increased expression of α1 (Ⅲ)procollagen mRNA by Northern blot analysis was the most predominant one among the threemRNAs during fibrogenesis. However, theenhanced expression of al (Ⅳ) procollagenmRNA occurred very early while the expressionof α1 (Ⅰ) mRNA was not enhanced much until themiddle stage of the exPeriment. D6smin (Dm)positive hepatic stellate cells (HSCs) and fewmyofibroblasts (MFs) in and around the necrotic areas expressed α1 (Ⅰ), α1 (Ⅲ) and α1 (Ⅳ)procollagen mRNA signals detected by in situhybridization at the early stage of theexperiment. All the three procollagen mRNAsignals thereafter mainly localized in fibroblasts (Fbs) and MFs in fibrotic septa during the middle and late stages of fibrosis, which distributedparallel to the corresponding collagens detected by immunohistochemical study. ln addition, the endothelial cells of sinusoids and the small blood vessels within the septa also showed α1(Ⅳ) procollagen mRNA and type Ⅳ collagen expressionCONCLUSION It is considered that "HSC-MF-Fb" effect cell system is the major cellularsource of collagen production in liver fibrosis, in which HSCs are collagen producing precursor cells in the early liver fibrogenesis, thereafter the synthesis of type Ⅰ, Ⅲ and Ⅳ collagens (Col Ⅰ, Col Ⅲ and Col Ⅳ) mainly derives fromMFs and Fbs, which play a very important role in the progress of liver fibrosis. The endothelialcells along sinusoids, as another source of

关键词： procollagen mRNA immunohistochemistry Northern blot analysis in situ hybridization liver fibrosis

Clinical Observation on the Long-Term Therapeutic Effects of Traditional Chinese Medicine for Treatment of Liver Fibrosis

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Journal of Traditional Chinese Medicine 2000年第4期20卷 247-250页

作者：杨宏志陈琰碧许瑞云沈伟生陈国庆刘煜 The Third Hospital Affiliated to Sun Yat-sen University of Medical Sciences Guangzhou 510630

48 patients with liver fibrosis due to hepatitis B were treated for 2 years with the drugs for tonifying the kidney, supplementing qi, cooling and invigorating the blood and detoxification. The symptoms were markedly improved, and serum ALT and bilirubin were recovered and kept normal in most of the cases. The mean levels of serum hyaluronic acid, procollagen peptide III and circulating immune complex were decreased and returned to normal after the treatment. B-ultrasonography showed that the portal vein kept in normal size in 82% of the patients, the enlarged portal vein diminished in diameter, and the enlarged spleen reduced.

关键词： Adult Antigen-Antibody Complex Biological Markers Drugs, Chinese Herbal Female Hepatitis B, Chronic Humans Hyaluronic Acid Liver Cirrhosis Male Middle Aged Peptide Fragments Portal Vein procollagen

Changes in myocardial collagen content before and after left ventricular assist device application in dilated cardiomyopathy

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Chinese Medical Journal 2004年第3期117卷 401-407页

作者：梁红 Johannes Müller 翁渝国 Gerd Wallukat 付平林汉生 Sabina Bartel Christoph Knosalla Reinhard Pregla Roland Hetzer Department of Cardiovascular and Thoracic Surgery Medical College of Jinan University Deutsches Herzzentrum Berlin Max-Delbrück Centre for Molecular Medicine Department of Cardiovascular Surgery Institute of Cardiovascular Diseases Tongji Medical University Department of Medical Statistics Medical College of Jinan University

BACKGROUND: The purposes of this study were to confirm the changes in myocardial collagen level after left ventricular assist device (LVAD) support in dilated cardiomyopathy (DCM), find the relation between these changes and prognosis, and test a practical method to assess the level of myocardial collagen. METHODS: Left ventricular samples were collected from DCM patients with different prognosis (transplanted group n = 8, weaning group n = 10) at the time when the LVADs were implanted and again during cardiac transplantation (n = 8). The level of neutral salt soluble collagen (NSC) and acid soluble collagen (ASC) was measured by Sircol collagen assay, and that of total collagen and insoluble collagen (ISC) by quantification of hydroxyproline (Hyp). Serum samples were collected from a portion of these patients (transplanted group, n = 6; weaning group n = 7) at the time the LVADs were implanted, 1 month after implantation and on explantation. Circulating concentration of carboxy-terminal propeptide of type I procollagen (P I CP), amino-terminal propeptide of type I procollagen (P I NP), amino-terminal propeptide of type III procollagen (P III NP) and type I collagen telopeptide (I CTP) were measured by the equilibrium type radioimmunoassay. RESULTS: Before LVAD implantation the level of NSC and ISC in the weaning group was higher but ASC in the transplanted group was lower than in the controls (P

关键词： Heart-Assist Devices Adult Cardiomyopathy, Dilated Collagen Comparative Study Female Heart Transplantation Humans Hydroxyproline Male Middle Aged Myocardium procollagen Prognosis

Inhibiting effect of antisense oligonucleotides phosphorthioate on gene expression of TIMP-1 in rat liver fibrosis

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World Journal of Gastroenterology 2001年第3期7卷 363-369页

作者： Qing He Nie Yong Qian Cheng Yu Mei Xie Yong Xing Zhou Yi Zhan Cao The Center of Infectious Disease Diagnosis and Treatment of PLA,Tangdu Hospital,Forth Military Medical University,Xi’an 710038,Shaanxi Province,ChinaDr,Qing He Nie graduated from Qinghai Medical College as a doctor in 1983,got master degree at Beijing 302 Army Hospital in 1993,got doctor degree at the Third Military Medical University in 1998,engaged in postdoctoral research at the Fourth Military Medical University from 1998 to 2000,now an associate professor,specialized in clinical and experimental research of infectious diseases,had more than 90 papers published,coauthor of ten books,first author of one book. The Center of Infectious Disease Diagnosis and Treatment of PLA Tangdu Hospital Forth Military Medical University Xi'an 710038 Shaanxi Province China. nieqinghe@***

AIM: To observe the inhibition of antisense oligonucleotides (asON) phosphorthioate to the tissue inhibitors metalloproteinase-1 (TIMP-1) gene and protein expression in the liver tissue of immunologically induced hepatic fibrosis rats. The possibility of reversing hepatic fibrosis through gene therapy was observed. METHODS: Human serum albumin (HSA) was used to attack rats, as hepatic fibrosis model, in which asONs were used to block the gene and protein expressing TIMP-1. According to the analysis of modulator, structure protein, coding series of TIMP-1 genome, we designed four different asONs. These asONs were injected into the hepatic fibrosis models through coccygeal vein. The results was observed by RT-PCR for measuring TIMP-1 mRNA expression, immunohistochemistry and in situ hybridization for collagen I, II, special staining of collagen fiber, and electron microscopic examination. RESULTS: Hepatic fibrosis could last within 363 days in our modified model. The expressing level of TIMP-1 was high during hepatic fibrosis process. It has been proved by the immunohistochemical and the electron microscopic examination that the asON phosphorthioate of TIMP-1 could exactly express in vivo. The effect of colchicine was demonstrated to inhibit the expressing level of mRNA and the content of collagen I, III in the liver of experimental hepatic fibrosis rats. However, the electron microscopy research and the pathologic grading of hepatic fibrosis showed that there was no significant difference between the treatment group and the model group (P> 0.05). CONCLUSION: The experimental rat model of hepatic fibrosis is one of the preferable models to estimate the curative effect of anti-hepatic fibrosis drugs. The asON phosphorthioate of TIMP-1 could block the gene and protein expression of TIMP-1 in the liver of experimental hepatic fibrosis rats at the mRNA level. It is possible to reverse hepatic fibrosis, and it is expected to study a new drug of antihepatic fibrosis on t

关键词： Gene Therapy Animals Collagen Type I Collagen Type III Disease Models, Animal Female Gene Expression Hepatocytes Immunohistochemistry Liver Liver Cirrhosis Microscopy, Electron Oligonucleotides, Antisense procollagen RNA, Messenger Rats Rats, Wistar Research Support, Non-U.S. Gov't Tissue Inhibitor of Metalloproteinase-1

Effects of lower fluence pulsed dye laser irradiation on production of collagen and the mRNA expression of collagen relative gene in cultured fibroblasts in vitro

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Chinese Medical Journal 2006年第18期119卷 1543-1547页

作者： YU Hai-yan CHEN Da-fang WANG Qi CHENG Hao Department of Dermatology Sir Run Run Shaw Hospital of Zhejiang University Hangzhou 310016 China

Background Lower fluence of 585-nm flashlamp-pumped pulsed dye laser has been successfully used as a nonablative technique in the treatment of wrinkles. The objective of this study was to evaluate the effect of the pulsed dye laser （585 nm） on the production of collagen and the mRNA expression of collagen related gene in fibroblasts in vitro. Methods Cultured fibroblasts were treated with a 585-nm flashlamp-pumped pulsed dye laser （ fluence 3 J/cm^2, 4 J/cm^2, spot size 7 mm, pulse duration 450 12s）. The production of collagen and the mRNA expression of transforming growth factor （TGF）-β1, SMAD2, SMAD3, SMAD4, SMAD7 and type Ⅰ procollagen α1, α2 in fibroblasts were investigated by colorimetry or real time polymerase chain reaction. Results The production of collagen was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm^2 （P 〈0.001）. The mRNA expression of TGF- β1, SMAD2, SMAD3, SMAD4, SMAD7 and procollagen I was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm^2 （P 〈0.001）. No significant difference of mRNA expression of SMAD2, SMAD3, SMAD4, SMAD7 and type Ⅰ procollagen was found between controls and fibroblasts treated with pulsed dye laser with a fluence of 4 J/cm^2 （P 〉0.05）. Conclusions Lower fluence （3 J/cm^2） pulsed dye laser increased the collagen production in fibroblasts by up-regulating TGF-β1, SMAD2, SMAD3, SMAD4, SMAD7 and type Ⅰ procollagen mRNA expression. These may be the reason it can be effectively used in the treatment of wrinkles.

关键词： pulsed dye laser fibroblasts procollagen SMADs