The most common forms of treatment for amblyopia are occlusion therapy and pharmacologic penalization. But these methods can not recover all the visual deficits of amblyopes and there are some disadvantages of the treatment which need to be considered, including adverse effects, compliance and social stigma. Recently, some novel approaches are investigated for improving the effectiveness and compliance of treatment for amblyopia.

AIM: To explore the molecular mechanisms about fibrosis and transforming growth factor (TGF) - β1 as well as inflammation in rats heart after acute myocardial infarction (AMI). METHODS: AMI model in rats was produced by left coronary artery ligation. Samples of rat cardiac tissue were collected at the end of 1 week, 4 weeks and 8 weeks. Hemodynamics had been performed before rats were sacrificed. Reverse transcription polymerase chain reaction (RT- PCR) and immunohistochemical methods were used to analyze mRNA expression and protein production of TGF- β1, respectively. Hydroxyproline was determined by chloramines T method. HE staining was resorted to analyze pathological myocardium after AMI. RESULTS: There were remarkable differences in hemodynamics between AMI groups and sham group (P＜0.01). Compared with sham group, TGF-β1mRNA expression and protein production and hydroxyproline quantification were enhanced greatly. Among them, the levels of TGF -β1 and hydroxyproline at 1 week were higher than those at 4 weeks or 8 weeks. A positive correlation between TGF- β1 protein and hydroxyproline was presented (r=0.75 - 0.99, P＜0.05). In microscope, leucocytes infiltrated significantly in the infarcted and border myocardium at 1 week after AMI, and were rarely seen at 4 weeks and 8 weeks. TGF - β1 protein were detected in cytoplasm of cardiac myocyte and leucocytes at 1 week, and at 4 and 8 weeks in myofibroblast and interstitial. CONCLUSIONS:TGF- β1 is upregulated and found in cytoplasm of cardiac myocyte and leucocytes as well as myofibroblast in heart after AMI,which is associated with dynamic changes of hydroxyproline content and inflammation. TGF - β1 is showed to play an important role in myocardial inflammatory repair and ventricular remodeling after AMI.

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

AIM: The study explored the significance of the imbalance of Th1/Th2 function after acute myocardial infarction (AMI). METHODS: Peripheral blood mononuclear cells from 33 AMI patients, 22 unstable angina (UA) patients and splenocytes from 35 AMI rats were collected. Cytokine-producing Th cells were monitored by 3-color flow cytometry after stimulated with phorbol myristate acetate (PMA) and ionomycin. IFN-? and IL-4 mRNA in the rat myocardium and chemokine receptors CCR3, CCR5 and CXCR3 mRNA on the surface of rat T lymphocytes after AMI were measured by RT-PCR. RESULTS: Th1 associated cytokine IFN-? significantly increased in patients with AMI and UA within 24 hours after the onset of symptom. the high ratio of IFN-?-producing T cells lasted short in patients with UA and recovered 1 week after the onset. In AMI patients, the high ratio of IFN-?-producing T cells could be examined 1 week and even 1 month after the onset. There was no significant difference on the frequencies of IL-4-producing peripheral T cells between each group. 1 week, 2 weeks and 1 month after AMI, IFN-? mRNA increased in the myocardium of rats, but there was no significant change on cytokine-producing Th cells and chemokine receptors on the surface of rat T lymphocytes. CONCLUSIONS: The Th1/Th2 functional imbalance and up-regulation of Th1 cell-functions exist after AMI and perhaps participate in the onset of AMI. Th1/Th2 functional imbalance may participate in the immune-mediated myocardial injury and ventricular remodeling after AMI as one of the pathogenesises of autoimmune disease.

Objective To explore the roles of autoantibodies against ?1 adrenoceptor(?1-receptor)and M2 cholinergic receptor(M2-receptor)in patients with chronic renal insufficiency.Methods The epitopes of the second extracellular loop of ?1 receptor and M2 receptor were synthesized and used respectively to detect the sera autoantibodies against ?1 receptor and M2 receptor by enzyme linked immunosorbent assay(ELISA) in 76 patients with chronic renal insufficiency,60 cases with hypertension and 40 healthy controls.Results In patients with chronic renal insufficiency,the positive rates of the autoantibodies against ?1-receptor and M2-receptor were 56.7% and 38.1% respectively,which were much higher than those of patients with hypertension(18.3% and 11.7%) and higher than those of healthy controls(17.5% and 15.0%)(all P

Objective To explore the role of the autoantibodies against ?_1-adrenergic receptor(?_1-receptor)in the development of hypertension with renal failure.Methods The epitopes of the second extracellular loop of ?_1-receptor were synthesized and used respectively to screen sera autoantibodies from patients with hypertension with renal failure(61 cases),hypertension without renal failure(58 cases) and healthy blood donors(40 cases,control) by ELISA method.Results The positive rates of the autoantibodies ?_1-receptor(69%,42/61) in patients with hypertension with renal failure were higher than those of patients with hypertension without renal failure(19%,11/58) respectively(P

To observe the relationship between positive rate of β1-adrenergic and AT1 receptors autoantibodies with serum concentration of cystatin C in 371 patients with diabetic nephropathy patients,107 patients with type 2 diabetes,and 47 subjects as healthy control.In patients with diabetic nephropathy,the positive rates of the β1 and AT1 receptors autoantibodies were significantly higher than those in patients with type 2 diabetes and normal controls.The titers of β1 and AT1 receptors autoantibodies in diabetic nephropathy patients with abnormal cystatin C were significantly higher than those with normal cystatin C concentration.These findings suggested that β1 and AT1 receptors autoantibodie may play important roles in the pathogenesis of diabetic nephropathy.

AIM: To clarify the relationship between the cytokine and collagen in myocardial remodeling after acute myocardial infarction(MI) in rats.METHODS: In MI group,Wistar rats were undergone acute myocardial infarction by ligation of the anterior descending coronary artery.Sham operation was made in rats as control.The mRNA expression of collagen and cytokines such as TNF-? and TGF-?_1 in infract and non-infarct region of left myocardium were detected by RT-PCR at different time point(3 d,1 and 4 weeks).RESULTS: Collagen type Ⅰ and Ⅲ elevated as well as the TNF-? and TGF-?_1 in the MI group at 3th day.Expression of collagen type Ⅰ and Ⅲ were higher in the infarct region than that in the non-infarct region even at 4 weeks.TNF-? and TGF-?_1 peaked at 1 week and declined gradually to the baseline,which was still higher than those in control group(P

To observe the dynamic changes of the TGF-beta1 expressed in the infarct and non-infarcted region of rat heart during the ventricular remodeling (day 3, 7, 28, 180), myocardial infarction rat model was made and relationship between the cytokine and indicator of myocardial remodeling was analyzed. After the detection of hemodynamic parameter was performed by the Powerlab devices, the size of myocardial infarction and the morphology change was detected by TTC and HE, respectively. The relative levels of mRNA of TGF- beta1, collagen type I, III, and fetal gene beta-MHC were detected by RT-PCR. The distribution of TGF- beta1 protein in the myocardium was detected by immunohistochemistry. The results showed that the size of infarction was higher than that of the sham operated groups in the infarcted group (44.5 +/- 0.5 vs 0). The difference in hemodynamic parameters between the infarcted group and sham operated group was significant (P < 0.01). HE staining showed that inflammatory cells were accumulated in the infarcted region at the beginning of the 3rd day, which lasted 4 weeks. Then, it decreased gradually. beta-MHC in the non-infarcted region rose from the 3rd day, reaching its peak at the 4th week, and it decreased gradually. The ratio of the collagen type I/III showed similar changes as compared with the sham operated groups (P < 0.01). And the relative mRNA levels in the non-infarcted group were significantly higher than that in the infarcted and sham operated group (P < 0.01) at day 180. Linear regression analysis indicated that the TGF-beta1 was positively correlated with the ventricular remodeling. It was concluded that the cytokine TGF-beta1 participates in the process of the myocardial remodeling, which could be a strategy in the interference of myocardial remodeling.

OBJECTIVEExposure to high concentrations of oxygen in the neonatal period may impair lung growth and is a major contributing factor to the development of bronchopulmonary dysplasia (BPD). Cell death from hyperoxic injury may occur through either an apoptotic or nonapoptotic pathway. The aim of the present study was to investigate the effect of hyperoxia on caspase-3 and p53 gene expression and apoptosis in the lungs of neonatal rats, so as to determine the type of cell death that occurs in the lungs of neonatal rats exposed to hyperoxia.

METHODSHyperoxic lung injury model was established by exposing to 95% O(2) in the neonatal period of Spraque-Dawley rats. The levels of caspase-3 mRNA and p53 mRNA expression in the lungs of neonatal rats exposed to 95% hyperoxia or room air were detected by RT-PCR. To quantify PCR products, PCR products were electrophoretically separated with 1.5% agarose gels. The optical density (A) values of the DNA bands were quantified by complete gel documentation and analysis system. The A ratios of p53/beta-actin denoted the relative content of p53 mRNA, results were showed as mean +/- standard deviation. The specific positive or negative bands of caspase-3 in electrophoresis gels were counted, Fisher's exact test of propabilities was used to determined statistically significant differences between two groups. We determined the extent of apoptosis taking place in the lungs of neonatal rats exposed to 95% hyperoxia using terminal deoxyribonucleotide transferase-mediated deoxyuridine triphosphate-fluorescence nick-end labeling (TUNEL) in 7-d-old neonatal lung. Under light microscope, five areas of lung parenchyma were systematically and randomly photographed from each animal and positive cells among 500 lung cells were calculated. Results were showed as mean +/- standard deviation.

RESULTSWe found increased levels of p53 messenger RNA, a gene associated with apoptosis, in the lungs of neonatal rat born and raised in 95% hyperoxia. Moderate increase in the level of p53 mRNA was found in the hyperoxic-treated group at 24 h (q = 3.2305, P > 0.05). Significant increase in the level of p53 mRNA was found in the hyperoxic-treated group at 48 h (q = 7.2941, P < 0.01). The levels of p53 mRNA expression in neonatal rat lungs exposed to 95% O(2) for 72 h or 96 h returned to normal level. The levels of caspase-3 mRNA expression were very low or absent in the hyperoxic-treated groups at 12 h, 24 h, 48 h, 72 h and 96 h or in the air-breathing groups at 12 h, 24 h, 48 h, 72 h and 96 h. An increase in the number of cells undergoing apoptosis was found in the hyperoxic-treated group at 7 d (F = 56.5010, P < 0.001) which was significantly greater than the number of apoptotic cells found in the lungs of rats of the same age exposed to room air.

CONCLUSIONOur results suggested that 95% hyperoxia could temporarily up-regulate the gene expression of p53, which induced the transcription of p21(WAF/CIP1) mRNA. Furthermore, p21(WAF/CIP1) could lead to cell cycle arrest and inhibit proliferation of lung cells. Meanwhile, p53 could also promote apoptosis of lung cells. Therefore, exposure to high concentrations of oxygen in the neonatal period may impair lung growth and is a major contributing factor to the development of bronchopulmonary dysplasia (BPD), and hyperoxia may affect the future lung growth and lead to barrier of lung development. The treatments of anti-apoptosis and promoting alveoli growth hold a promising perspective in hyperoxic lung injury. The level and ratio of caspase-3 gene expression were very low or absent in the lungs of neonatal rats exposed to 95% O(2) or room air. We speculated that caspase-3 gene expression was not essential in the hyperoxia induced lung cell apoptosis in neonatal rats.

Animals ; Animals, Newborn ; Apoptosis ; Caspase 3 ; genetics ; metabolism ; Disease Models, Animal ; Hyperoxia ; metabolism ; pathology ; In Situ Nick-End Labeling ; Lung ; metabolism ; pathology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism