Animals ; Humans ; Leptin ; genetics ; physiology ; Liver Cirrhosis ; etiology ; RNA, Messenger ; analysis ; Receptors, Cell Surface ; analysis ; physiology ; Receptors, Leptin ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

PURPOSE: To observe the changes of gait behavior and the expression of wound healing factors of transforming growth factor-β1 (TGF-β1), TGF-β3 and cAMP response element binding protein-1 (CREB-1) during the healing of Achilles tendon in a rat model, and to investigate whether gait analysis can be used to evaluate the tendon healing. METHODS: Achilles tendon of 40 healthy male Sprague-Dawley rats were transected and sutured to establish the Achilles tendon injury (ATI) model. They were randomly divided into 4 groups based on the observational time point at 1, 2, 4 and 6 weeks after injury (n = 10 for each group). Before modeling, 9 rats were randomly selected for CatWalk gait analysis, which contained step cycle, single stance time and average speed. Data were recorded as the normal controls. After then, ATI models were established in the left hind limbs of the all 40 rats (ATI group), while the right hind limbs were only cut and sutured without injury of the Achilles tendon (sham operation group). At 1, 2, 4 and 6 weeks after injury, the gait behavior of the corresponding group of rats (n = 9) as observed and recorded by CatWalk platform. After then, the rats were sacrificed and Achilles tendon of both limbs was harvested. The tendon healing was observed by gross anatomy and histological examination, and the protein and mRNA expression of TGF-β1, TGF-β3, CREB-1 were observed by immunohistochemistry and qPCR. The results of tendon gross grading were analyzed by Wilcoxon rank sum test, and other data were analyzed by one-way analysis of variance among multiple groups. RESULTS: Compared with normal controls, all gait indexes (step cycle, single stance time and average speed) were greatly affected following ATI, which however improved with time. The step cycle was significantly lower at 1, 2 and 4 weeks after ATI (compared with normal controls, all p < 0.05), but almost returned to the normal level at 6 weeks ((0.694 ± 0.102) vs. (0.503 ± 0.094) s, p > 0.05). The single stance time of the ATI group was significantly shorter at 1 and 2 weeks after operation ((0.078 ± 0.010) s at 1 week, (0.078 ± 0.020) s at 2 weeks, all p < 0.001) and revealed no significant difference at 4 weeks (p = 0.120). The average speed of ATI group at 1, 2, 4, 6 weeks was significantly lower than that in the normal control group (all p < 0.001). Gross observation showed that the grade of local scar adhesion in ATI group increased significantly at 2, 4 and 6 weeks, compared with the sham operation group (all p < 0.001). Extensive adhesion was formed at 6 weeks after ATI. The results of HE staining showed that the number of fibroblast increased gradually and arranged more orderly in ATI group at 1, 2 and 4 weeks (all p < 0.001), and decreased at 6 weeks, but it was still significantly higher than that of the sham operation group (p < 0.001). Immunohistochemistry showed that the positive expression of TGF-β1, TGF-β3, CREB-1 in ATI group was higher than that in the sham operation group at 4 time points (all p < 0.05), which reached the peak at 2 weeks after operation and decreased at 4 weeks (p = 0.002, p < 0.001, p = 0.041, respectively). The results of qPCR suggested that the mRNA expression of TGF-β1, TGF-β3, CREB-1 in ATI group was higher than that in the sham operation group at all-time points (all p < 0.05), which reached the peak at 2 weeks after operation, decreased at 4 weeks, and significantly decreased at 6 weeks (all p < 0.001). CONCLUSION Gait behavior indexes are associated with Achilles tendon healing. The study gives an insight of TGF-β1, TGF-β3, CREB-1 changes in the coursing of Achilles tendon healing and these cytokines may be able to be used to regulate the Achilles tendon healing.
Achilles Tendon ; Animals ; CREB-Binding Protein ; Gait Analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1/genetics* ; Transforming Growth Factor beta3 ; Wound Healing

OBJECTIVETo analyze the clinical features and TGFBI gene mutation in a Chinese family affected with Reis-Bucklers corneal dystrophy.

METHODSGenomic DNA was extracted from 53 members including 9 patients from the family. The 17 exons and splice region of introns of the TGFBI gene were amplified by PCR and directly sequenced. All family members were subjected to ophthalmologic examination.

RESULTSA heterozygous mutation (R124L) was found in exon 4 of the TGFBI gene among all patients from the family. The same mutation was not found among unaffected family members. The inheritance pattern of the family was identified as autosomal dominant, and the Reis-Bucklers corneal dystrophy in the family was diagnosed as the geographic type.

CONCLUSIONThe R124L mutation of the TGFBI gene probably underlies the pathogenesis of Reis-Bucklers corneal dystrophy in this Chinese family. Molecular genetic approach is useful for the proper diagnosis of this type of corneal dystrophy.

Corneal Dystrophies, Hereditary ; etiology ; genetics ; Female ; Humans ; Male ; Mutation ; Sequence Analysis, DNA ; Transforming Growth Factor beta1 ; genetics

OBJECTIVETo explore the expressions of transforming growth factor-beta1 (TGF-beta1) and its signaling transduction molecule Smad 2 and their significance in the development of glomerulosclerosis.

METHODSUsing in situ hybridization and immunohistochemistry to detect Smad 2 mRNA expression and TGF-beta1, collagen IV, fibronectin expression in renal biopsies from 61 cases with a spectrum of glomerulonephritis including IgA nephropathy (40 cases), membranous glomerulonephritis (10 cases) and sclerosing glomerulonephritis (11 cases), compared with 11 cases of glomerular mild lesion with image analysis system.

RESULTSWith the exception of Smad 2 mRNA expression in mild type IgA nephropathy, all other types of diseased glomeruli showed increased expression of both TGF-beta1 and Smad 2 mRNA when compared with the 11 cases of mild glomerular lesions. The expressions of glomerular TGF-beta1 and Smad 2 mRNA positively correlated with collagen IV and fibronectin deposition in the glomeruli.

CONCLUSIONSTGF-beta1 and Smad 2 may be involved in the excessive deposition of glomerular extracellular matrix and play an important role in the development of glomerulosclerosis.

Collagen Type IV ; analysis ; DNA-Binding Proteins ; genetics ; Fibronectins ; analysis ; Glomerulonephritis ; metabolism ; Humans ; Immunohistochemistry ; Kidney Glomerulus ; chemistry ; RNA, Messenger ; analysis ; Smad2 Protein ; Trans-Activators ; genetics ; Transforming Growth Factor beta ; analysis ; Transforming Growth Factor beta1

OBJECTIVETo elucidate the effects of exogenous basic fibroblast growth factor (bFGF) on biological characteristics of rat osteoblasts cultured in vitro.

METHODSThe osteoblasts isolated from a Sprague-Dawley rat and cultured in vitro were treated with different concentrations of bFGF (5-50 ng/ml) respectively. At 24 hours after treatment, the proliferating cell nuclear antigen was measured with immunocytochemistry, alkaline phosphatase (ALP) activity was determined and the expression of transforming growth factor beta 1 (TGF-beta(1)) was detected to observe the effects of bFGF on growth and differentiation of osteoblasts.

RESULTSbFGF (5-50 ng/ml) could obviously promote the growth of osteoblasts. The intracellular expression of TGF-beta(1) mRNA increased significantly, but the intracellular ALP content decreased.

CONCLUSIONSbFGF can obviously stimulate the proliferation of osteoblasts and promote the synthesis of TGF-beta(1), but cannot promote the differentiation of osteoblasts.

Alkaline Phosphatase ; metabolism ; Animals ; Cells, Cultured ; Fibroblast Growth Factor 2 ; pharmacology ; Osteoblasts ; drug effects ; metabolism ; Proliferating Cell Nuclear Antigen ; analysis ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

BACKGROUND/AIMS: The genetic polymorphism of transforming growth factor-beta1 (TGF-beta1) at codons 10 and 25 which influences the production of TGF-beta1 is related to fibrogenesis in the lung and liver. We evaluated the genetic polymorphism at codons 10 and 25 in controls and in patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC). METHODS: Blood samples were collected from controls (n=35), patients with LC (n=64), and HCC (n=49). Genomic DNA was isolated and polymerase chain reaction (PCR) was done for a segment including codons 10 and 25. The results of direct sequencing for PCR products were compared between the controls and the patients. RESULTS: There was no genetic polymorphism at codon 25 and three types of genetic polymorphism at codon 10. The leucine homozygous genotype (CTG/CTG) at codon 10 was more common in patients with LC than the controls (p=0.01) and especially in patients with LC caused by HBV (p=0.004). The polymorphism at codons 10 in patients with HCC was similar to the controls. However, leucine homozygous genotype was more common in patients with HCC of uninodular morphology than those of massive morphology (p=0.007). CONCLUSIONS: The genetic polymorphism of TGF-beta1 at codon 10 might be associated with LC and morphology of HCC. The potential usefulness of TGF-beta1 genotyping needs further studies in large scale.
Adult ; Aged ; Carcinoma, Hepatocellular/*genetics ; Codon/genetics ; Female ; Genotype ; Humans ; Korea ; Liver Cirrhosis/*genetics ; Liver Neoplasms/*genetics ; Male ; Middle Aged ; *Polymorphism, Genetic ; Sequence Analysis, Protein ; Transforming Growth Factor beta/*genetics ; Transforming Growth Factor beta1

OBJECTIVETo study the feasibility, efficiency and any benefits of recruitment maneuver (RM) in the facilitation of lung repair during recovery from ALI in acute lung injury (ALI) model of young piglets.

METHODSThe piglet model of ALI was established by an intravenous injection of lipopolysaccharide (LPS). Twelve ALI piglets were randomly divided into two groups: conventional ventilation (CON) and RM with low tidal volume. Arterial blood gas, dynamic lung compliance (Cdyn) and systematic hemodynamics were monitored during the treatment. TGF-β1 levels in bronchoalveolar lavage fluid (BALF) and plasma were measured. The mRNA expression of TGF-β1 in the lungs was assessed by real time PCR. Lung tissue was examined for morphological changes.

RESULTSNo significant difference was observed in cardiac output and peripheral vascular resistance (PVR) between the two groups. The extravascular lung water index (ELWI) from 6 hrs after ALI inducement and the pulmonary vascular permeability index (PVPI) 8 hrs after ALI inducement in the RM group decreased significantly compared with the CON group. Cdyn in the RM group increased quickly 1 hr after ALI inducement, and there was a significant difference between the two groups (P<0.05). P/F (ratio of PaO2 to FiO2) in the RM group was significantly higher than in the CON group from 2 hrs after ALI inducement (P<0.05). Alveolar-to-arterial oxygen difference in the RM group was obviously lower compared with the CON group from 2 hrs after ALI inducement (P<0.05). The levels of TGF-β1 in plasma and BALF and the mRNA expression of TGF-β1 in the lung tissue were lower than in the CON group. Volume density of alveolar aeration in the RM group was significantly higher than in the CON group, and the injury score in the RM group was lower (P<0.05).

CONCLUSIONSRM can improve gas exchange and Cdyn in the treatment of piglets with ALI. RM is a safe and effective approach to alveolar recruitment and can alleviate ventilation induced lung injury.

Acute Lung Injury ; pathology ; physiopathology ; therapy ; Aging ; Animals ; Disease Models, Animal ; Hemodynamics ; Lung ; pathology ; Male ; RNA, Messenger ; analysis ; Swine ; Transforming Growth Factor beta1 ; analysis ; genetics

OBJECTIVETo screen the differentially expressed genes in hepatic stellate cells (HSC) treated with transforming growth factor beta 1 (TGFbeta1) by cDNA microarray technique, and to elucidate the molecular pathogenesis of liver fibrosis involving TGFb1.

METHODSTotal RNA was extracted from HSC treated with TGFbeta1 and PBS by trizol and reverse-transcribed to double strand cDNA templates. Transcription of cDNA probe with biotin-labeling was performed, and then the obtained cDNA was hybridized with human cDNA microarray. The results were imaged by an Agilent scanner, and the differentially expressed genes were analyzed with bioinformatics software.

RESULTSOne hundred seventy-seven differentially expressed genes were screened from 13824 targeting genes; 123 genes were up-regulated, including connective tissue growth factor, tubulin epsilon 1, collagen, type V, alpha2, catenin delta 2, cadherin 6, type 2, Smad3, mitogen-activated protein kinase 4, growth factor receptor-bound protein 7 and MAP kinase-interacting serine/threonine kinase 1; 54 genes were down-regulated, including TNF receptor-associated factor 4, interferon regulatory factor 7, interferon inducible protein p78, bone morphogenetic protein 7, matrix gla protein, serine proteinase inhibitor, interferon stimulated gene 2.0 x 10(4), death-associated protein 6, metallothionein 1H and superoxide dismutase 2; in addition, 8 genes with unknown functions were also found.

CONCLUSIONThe differentially expressed genes in HSC treated with TGFbeta1 were successfully screened by cDNA microarray technique. It revealed that the molecular pathogenesis of liver fibrosis involving TGFbeta1 was the result of co-regulation by multiple factors. This information might be of help in searching for new targets in gene therapy.

Animals ; Cells, Cultured ; Gene Expression Profiling ; Hepatic Stellate Cells ; Liver Cirrhosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Rats ; Transforming Growth Factor beta1 ; genetics

OBJECTIVETo identify the mutation in transforming growth factor-beta1 gene (TGF beta1) in a Chinese patient with Camurati-Engelmann disease(CED).

METHODSDenaturing high-performance liquid chromatography (DHPLC) analysis was performed on the whole seven coding exons and exon-intron boundaries, then the mutation was identified by direct sequencing.

RESULTSMutation screening of TGF beta1 in this patient revealed a heterozygous missense mutation R218H in exon 4.

CONCLUSIONThe identification of the mutation could provide essential data for subsequent therapy and genetic counseling.

Base Sequence ; Camurati-Engelmann Syndrome ; genetics ; China ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Transforming Growth Factor beta1 ; genetics ; Young Adult

Animals ; Collagen Type I ; genetics ; Connective Tissue Growth Factor ; Corneal Injuries ; Fibronectins ; analysis ; Immediate-Early Proteins ; analysis ; genetics ; physiology ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; analysis ; genetics ; physiology ; RNA, Messenger ; analysis ; Rabbits ; Signal Transduction ; physiology ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; analysis ; genetics ; physiology ; Transforming Growth Factor beta1 ; Wound Healing ; physiology