Objective To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cells (HKC).Methods Cultured HKC were divided into 3 groups: (1) negative control; (2) low dose CTGF treated (rhCTGF 2. 5 ng/ml); (3) high dose CTGF treated (rhCTGF,5. 0 ng/ml). Expression of ?-smooth muscle actin (?-SMA) and fibronectin(FN) mRNA were measured by RT-PCR. Indirect immunofluorescence and flow cytometry methods were used to assess the level of intracellular ?-SMA protein. Concentration of FN secreted into the media was determined by ELBA. Results Upon the stimulations of different concentrations of rhCTGF,expression of ?-SMA and FN mRNA increased markedly(P

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of alpha-smooth muscle actin (alpha-SMA) were assessed by indirect immuno-fluorescence, and the percentage of alpha-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of alpha-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of alpha-SMA were markedly stronger than that in negative controls. The percentages of alpha-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9%, 65.5% vs 2.4%, P<0.01). alpha-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).
Actins/metabolism ; Cell Differentiation/*drug effects ; Cells, Cultured ; Epithelial Cells/*cytology ; Immediate-Early Proteins/*pharmacology ; Insulin-Like Growth Factor Binding Proteins/pharmacology ; Intercellular Signaling Peptides and Proteins/*pharmacology ; Kidney Tubules/*cytology

Background The conventional therapies for childhood amblyopia mainly include optometry,occluding therapy,atropine application,fine eyesight training,instrument or regular comprehensive therapy,but the clinical effectiveness of each method had its limitation.Chinese herbs have been used to the treatment of childhood amblyopia,however,their effects are still in controversy.Objective This study was to assess the evidence of efficacy of Chinese herbs combined with moderate methods for childhood amblyopia.Methods The published papers of randomized controlled trials (RCT) and controlled clinical trials (CCT) of Chinese herbs combined with moderate comprehensive therapy for childhood amblyopia from 2000 to 2015 were searched in China National Knowledge Infrastructure (CNKI),Wanfang database and PubMed database with computer.The methodology quality of included papers was evaluated according to the criteria of Cochrane and advanced Jadad scale.Data synthesis was facilitated using Reviewer Manager 5.1 software.The overall effect,or odds ratio (OR) of the treatment was analyzed by fixed effective model.Results Thirty-three RCT or CCT papers meeting inclusion criteria were included with the total sample size 6 666,and matched demography was showed in 25 papers;referential diagnosis criteria and effective criteria were described in 29 papers and 30 papers,respectively.Advanced Jadad scale showed 6,3 and 1-2 scores in 1,2 and 30 papers,respectively.The evidence showed that the total effectiveness of treatment within the group receiving Chinese herbs combined with conventional methods was higher than that in conventional group,with a statistically significant difference between groups (polled fixed effects model [OR] =3.92,95％ confidence interval [CI]:3.30-4.64,Z=15.74,P＜0.01).Conclusions The total effectiveness rate of Chinese herbs combined with conventional methods for childhood amblyopia is superior to only conventional treatment,indicating that Chinese herbs is effective for childhood amblyopia.More well-designed RCTs are needed to confirm the effect.

Objective:Our aim is to investigate the relationship of VEGF-C,VEGF-D and lymph node metastasis in pancreatic cancer,and to elucidate the role and significance of the cancerous peripheral lymphatic tissue.Methods: The expression of VEGF-C and VEGF-D was assayed by means of immunohistochemistry in 30 pancreatic carcinomas.Results:The positive rates of VEGF-C、VEGF-D were 73% (22/30)、57% (17/30) respectively in pancreatic cancer.The expression of VEGF-C、 VEGF-D in cancerous invasive edge was significantly higher than that in the center of cancerous tissues.There was no correlation between the expression of VEGF-C、VEGF-Dand the site,differentiation,histology types.Ⅲ~Ⅳ stages of pancreatic cancer showed strong expressions of VEGF-C、VEGF-D than that Ⅰ~Ⅱ stages of pancreatic cancer.The positive lymph node in positive VEGF-C and VEGF-D group were higher than that in the negative group.Conclusion:VEGF-C and VEGF-D induced lymphangiogenisis in pancreatic cancer,promoted the tumor cell lymph metastasis.

AIM:To observe the expression of connective tissue growth factor(CTGF)in unilateral ureteral obstruction(UUO)rats,and to explore its pathogenic role in renal tubulointerstitial fibrosis.METHODS:48 Wistar rats were randomly divided into sham-operated and UUO group.The rats were sacrificed at day 1,3,7,and 14.The degree of tubulointerstitial damage was scored according to the Masson staining.The mRNA and protein levels of CTGF,transforming growth factor-?1(TGF-?1),collagen Ⅰ(Col Ⅰ),and plasminogen activator inhibitor-1(PAI-1)were detected by reverse transcriptional-polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively.Expression of CTGF protein in the kidney was also assessed using Western blotting.RESULTS:TGF-?1 mRNA level began to increase as early as 1 day after UUO.This increase was followed by the elevation of CTGF mRNA level,which began to increase at third days after UUO(P

Objective To study the effects of CD2-associated protein (CD2AP) on podocyte adhesion and extension ability and to explore its possible mechanism. Methods Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium at 33℃permissive conditions. The podocytes were transfected with CD2AP small interfering RNA (siRNA) and serambing sequences labeled with fluorescein were taken as control. The transfected podocytes were trypsinized and seed into collagen IV coated plates. The relative cell adhesion and cell area were examined 90 min later. Apoptotic rates of CD2AP siRNA transfected podoeytes and different PAN concentrations incubated podoeytes were detected by flow cytometer. The distribution of F-actin was observed under laser scanning confoeal microscope. Nephrin protein expression and its phosphorylation level were examined by immunofluorescence and Western blot. Results The relative ceil adhesion of CD2AP siRNA transfected podocytes was apparently lower than that of control group[(41.72±6.07)% vs (64.46±8.53)%, P<0.05]. The cell area analysis had the similar result. The apoptotic rate of CD2AP siRNA transfected podocytes was significantly higher than that of the controls [(5.73±0.61)% vs (3.26±0.45)%, P<0.05]. 100 mg/L PAN could markedly induce podocytes to apoptosis and impair cell adhesion ability (P<0.05). Nevertheless, no significant difference was found in cell body spreading (P>0.05). The distribution of F-actin in CD2AP depletion podocytes was apparently altered. The expression of nephrin protein and its phosphorylation level was conspicuously descended to some degree (P<0.05). Conclusions CD2AP depletion facilitates podocyte apoptosis and impairs cell adhesion function. Cytoskeleton confusion and nephrin signaling weakness caused by CD2AP depletion may he partly responsible for the decline of cell adhesion and spreading.

To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.
Fibrosis ; Kidney/*pathology ; Kidney Diseases/etiology ; Kidney Diseases/pathology ; Kidney Diseases/*prevention & control ; Mycophenolic Acid/*analogs & derivatives ; Mycophenolic Acid/*pharmacology ; Random Allocation ; Rats, Sprague-Dawley ; Ureteral Obstruction/*complications

Objective To study the value of the application of 320-row detector dynamic volume CT in diagnosis of complicated congenital heart diseases(CCHD).Methods Datas of 43 cases with CCHD,who examined by ECC-gated 320-row CT angiography (CTA)as preoperative examinations were collected.All the CT data were processed by combined application of multi-imaging post-processing technique to show the anatomic malformation based on segmental analysis,and were compared with the transthoracic ech-ocardiography (TTE )and surgical results .Results 1 97 various malformations were found in 43 cases and proved by operation, among which there were 87 intracardiac malformations,the diagnostic accuracy with CTA and TTE were 86.2%,89.7%,and there was no obvious difference between them(P >0.05 ).1 10 extracardiac malformations (including connection malformations between heart and vessels,extracardiovascular malformation,bronchial or other nonvascular structure malformation)were found ,the accura-cy with CTA and TTE were 91.8% and 76.4%,the former was superior to latter (P <0.05)Conclusion 320-row detector dynamic volume CT in diagnosis of CCHD has very important meaning and practical application value,should be used as the significant choice for clinical diagnosis of CCHD.

Through analyzing the research literature about the mechanism of acupuncture intervention on vision in recent years, this article reviewed the mechanism of acupuncture intervention on vision from the aspects of central processing of visual information, the influnce of information transmission in the retina, the changes in visual microcirculation, optic nerve and neurotransmitter transmission, visual electrophysiology, visual plasticity. This article also proposed the future research idea for this field.

AIM: To investigate the effect of high glucose concentration on serum and glucocorticoid induced protein kinase (SGK) mRNA and protein expressions in human proximal tubular epithelial cells (HKC) and the possible role of SGK in the production of extracellular matrix (ECM) of HKC under the condition of high glucose. METHODS: HKC was divided into 3 groups: control glucose group (CG group, 5 5 mmol/L D-glucose); high glucose group (HG group, 25 mmol/L D-glucose) and osmotic control group (MG group, 19 5 mmol/L mannitol and 5 5 mmol/L D-glucose). The expressions of SGK mRNA and protein were assessed by semi-quantitative RT-PCR and Western blotting respectively. The level of secretary and cytoplasmic fibronectin (FN) were detected by enzyme-linked immunoabsordent assay (ELISA) and indirect-immunofluorescence. RESULTS: HKC expressed SGK1, SGK2 and SGK3 at mRNA and protein levels. Their mRNA level were up-regulated since 2 hours after cells exposed to D-glucose and this up-regulation persisted to the end of 8th hour, and SGK1 protein level elevated simultaneously. On the other hand, the increased FN secretion by high glucose was in a time-dependent manner and its improved secretion threshold was just followed by the high expression of SGK1. CONCLUSIONS: In response to high glucose, the expression of SGK1, SGK2 and SGK3 in human proximal tubular epithelial cells were up-regulated which was accompanied with FN accumulation. The high expression of SGK may mediate overproduction of ECM in proximal tubular epithelial cells and contribute to the diabetic nephropathy.