OBJECTIVETo explore the possible effect of transforming growth factor-beta(1) (TGF -beta(1)) on the development of renal fibrosis in human mesengial proliferative glomerulonephritis (MsPGN).

METHODSImmunohistochemistry method, sirius red staining polarization microscopy and the computer imaging analysis system were used to detect the expression of TGF-beta(1), the distribution of collagen I, collagen III and collagen IV.

RESULTIn MsPGN with renal fibrosis, collagen IV was increased markedly,and collagen I and collagen III appeared in the expanded mesengial matrix abnormally. Collagen III and collagen IV were increased markedly in tubulointerstitium. TGF-beta(1) expression was positively correlated with the expression of collagen I, collagen III and collagen IV in tubulointerstitium (r=0.82 0.92,P<0.01), and negatively correlated with I/III, I/IV and III/IV (r=-0.83,-0.92, P<0.001).

CONCLUSIONAbnormal increase of TGF-beta(1) may be one of the important factors associated with glomerular sclerosis and tubulointerstitial fibrosis through the increment and abnormal distribution of collagen I, collagen III and collagen IV.

Collagen ; analysis ; Fibrosis ; Glomerulonephritis, Membranoproliferative ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Kidney ; pathology ; Transforming Growth Factor beta ; analysis ; Transforming Growth Factor beta1

OBJECTIVETo examine the expression of activin A (ACT A) and transforming growth factor-beta 1 (TGF-beta 1) during mandibular lengthening and elucidate the difference between the role of ACT A and TGF-beta 1 during mandibular distraction osteogenesis.

METHODSkeletally mature white new zealand rabbits were established right mandibular distraction osteogenesis model. The regenerating tissue of animals' lengthened mandibes were harvested at different time points to have immunohistochemistric research of ACT A, TGF-beta 1 protein and analysis ACT A, TGF-beta 1 mRNA by using RT-PCR semiquantitative mean.

RESULTSAT the end of latency period day, positive stain of ACT A were found in the osteoblasts while positive stain of TGF-beta 1 was found in mesenchymal cells. At the end of distraction phase, fibrosis tissue had no stain of ACT A, but had strong stain of TGF-beta 1. At the period of fixation days of 20 days, both cytoplasm of osteoblasts and extracellular matrix in primary mineralization front were strongly stained of ACT A. The osteoblasts, osteoid and osteocytes in peripheral new bone zone were moderately stained of ACT A. TGF-beta 1 had strongly positive stained in fibrosis zone and weekly positive stained in primary mineralization front and peripheral new bone zone. There were also broad activin A stains in cytoplasm of osteoblasts, osteoid and cytoplasm of ACT A, TGF-beta 1 in osteocytes after distraction for 30 days. Activin A mRNA began to express at the end of latency period. Expression for activin A mRNA increased gradually along with the beginning of distraction and at the peak in distraction of 10 days and 20 days, while TGF beta 1 mRNA increased at the peak at the end of latency period.

CONCLUSIONACT A and TGF beta 1 have different role during rabbit mandibular distraction osteogenesis.

Activins ; analysis ; physiology ; Animals ; Female ; Immunohistochemistry ; Inhibin-beta Subunits ; analysis ; physiology ; Mandible ; surgery ; Osteogenesis, Distraction ; Rabbits ; Transforming Growth Factor beta ; analysis ; physiology ; Transforming Growth Factor beta1

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 observe the effects of Genistein on TGF-beta1 expression and the intracellular free Ca2+ concentration in human hypertrophic scar fibroblasts, and to discuss the mechanism of the anti-fibrosis effect.

METHODSFibroblasts were derived from human hypertrophic scar tissue and cultured in vitro. Genistein in different concentrations (25, 50, 100 micromol/L) was administrated to the fibroblasts, respectively. After 48 hours of co-culture, the expression of TGF-beta1 mRNA and protein were examined by RT-PCR and Western-Blot assay respectively. The intracellular free Ca2+ concentration in hypertrophic scar fibroblasts pretreated by Genistein was determined by laser confocal scanning microscopy with or without the stimulation of bFGF.

RESULTSGenistein inhibited the expression of TGF-beta1 in hypertrophic scar fibroblasts on a concentration-dependent manner. bFGF significantly elevated the intracellular free Ca2+ concentration, however its stimulating effect was remarkably alleviated when the fibroblasts were pre-treated by Genistein.

CONCLUSIONSGenistein can reduce the expression of TGF-beta1 and block the accumulation of intracellular free calcium induced by growth factors. It maybe one of the possible mechanisms of Genistein's antifibrosis effect.

Calcium ; analysis ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Genistein ; pharmacology ; Humans ; Transforming Growth Factor beta1 ; metabolism

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 investigate the clinical significance of transforming growth factor-beta 1 (TGF-β1) in children with primary IgA nephropathy (IgAN).

METHODSThirty children who were diagnosed with primary IgAN by renal biopsy between May 2008 and October 2012 were included in the study. Thirty healthy children were used as the control group. Urinary and blood TGF-β1 levels were measured using enzyme-linked immunosorbent assay, and the protein expression of TGF-β1 in the renal tissue was measured by immunohistochemistry. The correlation between TGF-β1 levels in blood, urine, and renal tissue and their relationship with clinical indices were analyzed.

RESULTSChildren with primary IgAN had significantly higher urinary and blood TGF-β1 levels than the control group (P<0.01). Urinary TGF-β1 level was positively correlated with the pathological grade of renal tissue (r=0.557, P=0.001), and a significant positive correlation was also found between the TGF-β1 expression in the renal tissue and the pathological grade of renal tissue (r=0.682, P<0.01). There was no correlation between TGF-β1 levels in blood and renal tissue (r=0.038, P=0.844).

CONCLUSIONSUrinary TGF-β1 level is significantly positively correlated with the pathological severity of disease in children with primary IgAN. Clinical measurement of urinary TGF-β1 may be of great practical value in predicting the progression and prognosis of chronic nephropathy.

Adolescent ; Child ; Female ; Glomerulonephritis, IGA ; metabolism ; pathology ; Humans ; Kidney ; chemistry ; pathology ; Male ; Transforming Growth Factor beta1 ; analysis ; physiology ; urine

OBJECTIVE: To observe the therapeutic effects of different waves of electroacupuncture (EA) on knee osteoarthritis (KOA), and to explore the mechanism of different waves of EA on promoting cartilage repair. METHODS: Ninety- seven patients with KOA were randomly divided into a dilatational wave group (32 cases, 2 cases dropped off), a continuous wave group (32 cases, 2 cases dropped off) and a discontinuous wave group (33 cases, 3 cases dropped off). The same acupoints of Xuehai (SP 10), Liangqiu (ST 34), Dubi (ST 35) and Neixiyan (EX-LE 4) were selected in the three groups. The dilatational wave (frequency of 2 Hz/10 Hz) was used in the dilatational wave group, the continuous wave (frequency of 10 Hz) was used in the continuous wave group, and the discontinuous wave (frequency of 10 Hz) was used in the discontinuous wave group. All the needles were retained for 30 min. All the treatment was given 3 times a week (on Monday, Wednesday and Friday) for 4 weeks. Lysholm knees scoring scale (LKSS) was used to evaluate the knee joint function before and after treatment, and the content of transforming growth factor-β1 (TGF-β1) in the joint effusion before and after treatment was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared before treatment, the total score and each score of LKSS were increased after treatment in the three groups (all <0.05). The improvements of total score, pain score, instability score, swelling score of LKSS in the continuous wave group and the dilatational wave group were superior to those in the discontinuous wave group (all <0.05). The content of TGF-β1 in the joint effusion in each group was increased after treatment (<0.05), and the improvement in dilatational wave group was superior to thoes in the continuous wave group and the discontinuous wave group (all <0.05). CONCLUSION The three different waves of EA could all improve the clinical symptoms of KOA, which may promote cartilage repair by increasing TGF-β1 content. The dilatational wave had the best overall effect, which can be used as a clinical optimal treatment.
Acupuncture Points ; Electroacupuncture ; Humans ; Knee Joint ; Osteoarthritis, Knee ; therapy ; Synovial Fluid ; chemistry ; Transforming Growth Factor beta1 ; analysis ; Treatment Outcome

OBJECTIVETo investigate the effect of lipopolysacharide (LPS) in different concentrations on the biological features and growth factor secretion power of U937 cell line.

METHODSIn vitro cultured U937 cells were stimulated by 0 (as control), 0.1, 1.0, 10.0, 50.0 and 100 micro g/ml LPS respectively for 24 hours. Thereafter, the cell proliferation ability was determined by MTT method. The cell apoptosis rate was determined by flow cytometry. The changes in the contents of transforming growth factor beta(1) (TGFbeta(1)) and vascular endothelial growth factor (VEGF) of the supernatant of the cell culture were assessed by ELISA.

RESULTSApoptosis and TGFbeta(1) secretion could be induced by LPS in dose of 0.1 to 100 micro g/ml when compared with that without LPS challenge (P < 0.05 - 0.01). In detail, LPS in lower dose (0.1, 1.0 and 10.0 micro g/ml) could promote the proliferation of U937 (P < 0.05 - 0.01) but exerted no effect on VEGF secretion. In contrary, LPS in high dose (50 and 100 micro g/ml) could promote VEGF secretion (P < 0.01) but exerted no effects on the proliferation of U937 cells.

CONCLUSIONU937 cells could be activated to increase the secretion of TGFbeta(1) by LPS in optimal dose of 0.1 - 10.0 micro g/ml, but the secretion of VEGF could only be promoted by LPS in higher concentration.

Apoptosis ; drug effects ; Cell Division ; drug effects ; Humans ; Lipopolysaccharides ; pharmacology ; Transforming Growth Factor beta ; analysis ; secretion ; Transforming Growth Factor beta1 ; U937 Cells ; drug effects ; secretion ; Vascular Endothelial Growth Factor A ; analysis ; secretion

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