Effects of rhBNP on myocardial fibrosis after myocardial infarction in rats

Purpose: We aimed to observe the effects and mechanism of rhBNP treatment on myocardial fibrosis (MF) after myocardial infarction (MI). Methods: SPF rats were separated into 3 groups: normal, MI (ligation of left coronary artery), and MI + rhBNP (recombinant human brain natriuretic peptide). Rats in MI + rhBNP group were given 30 μg/kg for 2 days before modeling and for 4 weeks after modeling. mRNA levels and the expression levels of TGF-β1 (transforming growth factor) and CTGF (connective tissue growth factor) in 3 groups were analyzed using the RT-qPCR and western blotting analysis, respectively. Furthermore, myocardial volume fraction (CVF) was analyzed using the Sirius Red F3B (SR) while the percentage of type I and III collagen in 3 groups were analyzed using the immunohistochemical staining. Results: Compared with the normal group, the levels of TGF-β1, CTGF, CVF, type I and III collagen were higher in MI group. However, mRNA levels of TGF-β1 and CTGF were significantly decreased in MI + rhBNP compared to MI groups. Expression of TGF-β1 was lower while that of CTGF was higher in MI + rhBNP group than that in MI group. Besides, CVF, and type I and III collagen were lower in MI + rhBNP group compared with MI group. Conclusion: rhBNP could significantly decrease the TGF-β1 and CTGF levels in post-MI so as to inhibit the type I and III collagen deposition in MF of post-MI. rhBNP will be benefit for the improvement of MF.     

Dual therapy of rosiglitazone/pioglitazone with glimepiride on diabetic nephropathy in experimentally induced type 2 diabetes rats

Diabetic nephropathy is a major cause of end-stage renal disease (ESRD) in the general population. It is estimated that diabetic nephropathy will eventually develop in about 40% of all patients with diabetes; therefore, prevention is critical for delaying the development and progression of diabetic kidney disease. Despite extensive efforts, medical advances are still not successful enough to prevent the progression of the disease. In the present study, we focused on the comparison of combination therapies and whether they offered additional renoprotection. Type 2 diabetes mellitus was induced by intraperitoneally administering streptozotocin (90 mg/kg) in neonatal rats and then these rats were treated with rosiglitazone (1.0 mg/kg) in combination with glimepiride (0.5 mg/kg) or with pioglitazone (2.5 mg/kg) in combination with glimepiride (0.5 mg/kg). Diabetic nephropathy markers were evaluated by biochemical and ELISA kits and renal structural changes were examined by light microscopy and transmission electron microscopy. Results show that the combination of pioglitazone with glimepiride is more effective in amelioration of diabetic nephropathy than rosiglitazone with glimepiride drug therapy due to glycemic control, suppressing albumin excretion rate, total protein excretion rate and augmented TNF-a signaling during the development of streptozotocin induced type 2 diabetic nephropathy.     

Fibrinogen-like protein 2 expression correlates with microthrombosis in rats with type 2 diabetic nephropathy

Fibrinogen-like protein 2 (fgl2),a novel prothrombinase,is involved in microthrombosis.We examined fgl2 expression in the glomerular and tubulointerstitial capillaries and its correlation with microthromsis in rats with streptozocin-induced type 2 diabetic nephropathy.Our RT-PCR and immunoblotting analysis showed that fgl2 mRNA and protein levels were increased in microvascular endothelial cells of the glomeruli and renal interstitia at week 19 and became significantly elevated with the development of diabetic nephropathy (P < 0.01).Fgl2 was not or only weakly expressed in the renal tissues of normal rats.Furthermore,a direct significant correlation (r=0.543,P < 0.01) was found between fgl2 expression and microthrombotic capillaries in the renal tissues.Enzyme linked immunosorbent assays (ELISA) additionally showed that circulating TNF-α levels in rats with type 2 diabetes were significantly elevated and closely correlated with fgl2 expression (r=0.871,P < 0.01).Our results suggest that fgl2 may activate renal microthrombosis,thus contributing to glomerular hypertension and renal ischemia.     

Reduced beta 2 glycoprotein I improves diabetic nephropathy via inhibiting TGF-β1-p38 MAPK pathway

Purpose: Beta 2 glycoprotein I (β2GPI) has been shown the positive effect on diabetic atherosclerosis and retinal neovascularization. β2GPI can be reduced by thioredoxin-1, resulting in the reduced state of β2GPI. The possible protective effects of β2GPI and reduced β2GPI on diabetic nephropathy (DN) are not fully elucidated. The purpose of this study was to test a hypothesis that β2GPI and reduced β2GPI would improve DN in streptozotocin (STZ) induced diabetic mice and high-glucose (HG) exposed rat mesangial cell (RMC). Methods: The STZ-induced Balb/c mice and HG exposed RMCs were administrated with β2-GPI and reduced β2-GPI at different time and concentrations gradient respectively. The changes of glomerular structure and expression of collagen IV, TGF-β1, p38 MAPK and phospho-p38 MAPK in renal cortical and mesangial cells were observed by immunohistochemical techniques, quantitative real-time PCR and western blot with or without the treatment of β2-GPI and reduced β2-GPI. Results: β2GPI and reduced β2GPI improved early clinical and pathological changes of DN in STZ-diabetic mice. Treatment with β2GPI and reduced β2GPI in the STZ-diabetic mice and HG exposed RMCs resulted in decrease expression levels of TGF-β1 and collagen IV, with concomitant decrease in phospho-p38 MAPK expression. Conclusions: β2GPI and reduced β2GPI improved renal structural damage and kidney function. The renoprotective and antifibrosis effects of β2GPI and reduced β2GPI on DN were closely associated with suppressing the activation of the TGF-β1-p38 MAPK pathway.     

厄洛替尼减轻STZ诱导的糖尿病肾病模型大鼠的肾损伤

目的:研究表皮生长因子受体(EGFR)抑制剂厄洛替尼(erlotinib)对糖尿病肾病大鼠肾脏的保护作用及机制。方法:采用大剂量(55 mg/kg)链脲佐菌素(STZ)腹腔注射诱导大鼠糖尿病肾病模型,以1周后血糖值16.7 mmol/L的大鼠为造模成功的标准。将糖尿病大鼠随机分为2组[STZ组和STZ+erlotinib(100 mg·kg~(-1)·d~(-1))组],并以正常大鼠为对照组(control组)。Erlotinib处理4周后,检测大鼠空腹血糖、血清肌酐和24 h尿蛋白含量的变化;HE染色和Masson染色观察肾脏组织病理学改变;Western blot检测各组肾脏组织中EGFR、p-EGFR、转化生长因子β1(TGFβ1)、Smad2/3、p-Smad2/3、Ⅳ型胶原蛋白(ColⅣ)和纤连蛋白(fibronectin)的蛋白水平;活性氧簇(ROS)和丙二醛(MDA)试剂盒分别检测各组肾脏组织中ROS和MDA水平。结果:与control组相比,STZ组血糖、24 h尿蛋白和血清肌酐水平均显著升高(P0.01),肾组织形态学出现异常变化;与STZ组相比,STZ+erlotinib组的血糖、24 h尿蛋白水平和血清肌酐水平显著降低(P0.05),肾小球结构恢复正常,肾小球系膜细胞增生程度明显减弱。厄洛替尼明显抑制了STZ大鼠肾组织中p-EGFR、TGFβ1、p-Smad2/3、ColⅣ和fibronectin蛋白水平,也明显抑制了STZ大鼠肾组织中ROS和MDA水平。结论:厄洛替尼可能通过抑制EGFR/TGFβ1-Smad2/3信号通路的激活来抑制糖尿病肾病肾组织的纤维化和氧化应激反应,从而减轻肾损伤。     

Effects of Smad decoy ODN on shear stress-induced atherosclerotic ApoE-/-mouse

Atherosclerosis is a complex disease which involves both genetic and environmental factors in its development and progression. Shear stress is the drag force per unit area acting on the endothelium as a result of blood flow, and it plays a critical role in plaque location and progression. TGF-β1 is often regarded to have pro-atherosclerotic effect on vascular disease. TGF-β1 downstream targets Smad, for regulating a set of genes associated with atherosclerosis. Therefore, modulation of TGF-β1 and Smad expression may be the important targets for the prevention and treatment of shear stress-induced vascular disease. However, the precise mechanism of the anti-atherosclerotic effects of novel therapeutic approach has not been elucidated by using animal models regarding the shear stress-induced vascular disease. Therefore, we designed to test whether Smad decoy ODN would prevent the development of atherosclerosis in the shear stress-induced ApoE-/-mice on a western diet. We examined the effect of cast placement on the development of atherosclerosis, and the carotid artery was harvested at the sacrifice to observe histological changes. Also, we evaluated the impact of Smad decoy ODN in the regulation of genes expression related to atherosclerosis, including TGF-β1, PAI-1, and α-SMA. Our results showed that western diet with cast placement developed atherosclerosis in ApoE-/-mouse. Also, administration of Smad decoy ODN decreases the expression of TGF-β1, PAI-1, and α-SMA. These results demonstrate the potential of Smad decoy ODN to prevent the progression of atherosclerosis in ApoE-/-mouse model with western diet and shear stress.     

白藜芦醇对糖尿病肾病大鼠醛糖还原酶的影响

目的:观察白藜芦醇对糖尿病肾病大鼠醛糖还原酶的影响。方法:将48只雄性SD大鼠随机分为空白对照组、模型组、白藜芦醇低、中、高剂量组和依帕司他组(n=8)。除正常对照组外,其余各组大鼠腹腔注射STZ(55 mg.kg-1),成功造模后白藜芦醇组分别给予5、154、5 mg.kg-1白藜芦醇,依帕司他组给予10 mg.kg-1依帕司他,连续灌胃6 w后,检测大鼠血糖、体重,HE染色检测肾脏病理改变,ELISA法检测醛糖还原酶(AR)活性。结果:与正常对照组相比,糖尿病组血糖与AR活性明显升高(P<0.05),与模型组比较,白藜芦醇各组血糖与AR活性明显降低(P<0.05)。结论:白藜芦醇可降低糖尿病大鼠血糖,其降糖效应可能与抑制AR活性有关。     

小檗碱对糖尿病肾病大鼠C反应蛋白的影响

目的:观察小檗碱对糖尿病肾病大鼠C反应蛋白的影响。方法:将48只雄性SD大鼠随机分为空白对照组、模型组、小檗碱75、150及300mg.kg-1组、罗格列酮组六组,每组8只。除正常对照组外,其余各组大鼠腹腔注射链脲佐菌素(STZ,55mg.kg-1),成功造模后小檗碱各组及罗格列酮组分别灌胃给予75、150、300mg.kg-1小檗碱以及4mg.kg-1罗格列酮,连续6w后检测大鼠血糖、体重,HE染色检测肾脏病理改变,ELISA法检测C反应蛋白活性。结果:与正常对照组相比,糖尿病组血糖与C反应蛋白活性明显升高(P<0.05),与模型组比较,小檗碱各组血糖与C反应蛋白活性明显降低(P<0.05),有统计学意义。结论:小檗碱可降低糖尿病大鼠血糖和C反应蛋白,减轻糖尿病肾病。     

Effects of hydrogen sulfide on high glucose-induced glomerular podocyte injury in mice

The aim of this study was to assess the effects of hydrogen sulfide on high glucose-induced mouse podocyte (MPC) injury and the underlying mechanisms. Mouse podocytes were randomly divided into 4 groups, including high glucose (HG), normal glucose (NG), normal glucose + DL-propargylglycine (PPG), and high glucose + NaHS (HG + NaHS) groups for treatment. Then, ZO-2, nephrin, β-catenin, and cystathionine γ-lyase (CSE) protein expression levels were determined by western blot. We found that high glucose significantly reduced nephrin, ZO-2, and CSE expression levels (P<0.05), and overtly elevated β-catenin amounts (P<0.05), in a time-dependent manner. Likewise, PPG at different concentrations in normal glucose resulted in significantly lower CSE, ZO-2, and nephrin levels (P<0.05), and increased β-catenin amounts (P<0.05). Interestingly, significantly increased ZO-2 and nephrin levels, and overtly reduced β-catenin amounts were observed in the HG + NaHS group compared with HG treated cells (P<0.01). Compared with NG treated cells, decreased ZO-2 and nephrin levels and higher β-catenin amounts were obtained in the HG + NaHS group. In conclusion,CSE downregulation contributes to hyperglycemia induced podocyte injury, which is alleviated by exogenous H₂S possibly through ZO-2 upregulation and the subsequent suppression of Wnt/β-catenin pathway.     

WNT4及其抑制因子SFRP1在糖尿病肾病大鼠肾组织中的表达

目的观察WNT4/β-catenin信号通路及其抑制因子分泌型卷曲相关蛋白1(SFRP1)在糖尿病肾病(DN)大鼠肾组织中的表达变化,探讨其在肾脏纤维化发生发展中的可能作用。方法将大鼠随机分为正常对照(NC)组和DN组,8只/组。尾静脉注射STZ 55 mg/kg复制IDDM模型。HE、PAS及Masson染色观察肾组织形态学结构和纤维化病变;免疫组织化学法观察WNT4和β-catenin蛋白在肾组织的表达部位;Western blot检测WNT4、SFRP1、β-catenin、p-GSK-3β、GSK-3β、CollagenⅠ、α-SMA和E-cadherin蛋白在肾组织中的表达;Real-time PCR检测WNT4及SFRP1 mRNA在肾组织中的表达。结果与NC组相比,DN组大鼠肾组织纤维化病变明显;WNT4蛋白和mRNA表达显著增多(P<0.05);β-catenin、p-GSK-3β、α-SMA和CollagenⅠ蛋白的表达显著增多(P<0.05);E-cadherin蛋白表达显著减少(P<0.05);SFRP1蛋白和mRNA表达显著下降(P<0.05)。结论在DN发病中,WNT4/β-catenin信号通路异常活化;SFRP1表达减少可能抑制该通路,促进了DN肾纤维化的发生发展。     

Biologic Response of Degenerative Living Human Nucleus Pulposus Cells to Treatment with Cytokines

Purpose

To investigate the molecular responses of various genes and proteins related to disc degeneration upon treatment with cytokines that affect disc-cell proliferation and phenotype in living human intervertebral discs (IVDs). Responsiveness to these cytokines according to the degree of disc degeneration was also evaluated.

Materials and Methods

The disc specimens were classified into two groups: group 1 (6 patients) showed mild degeneration of IVDs and group 2 (6 patients) exhibited severe degeneration of IVDs. Gene expression was analyzed after treatment with four cytokines: recombinant human bone morphogenic protein (rhBMP-2), transforming growth factor-β (TGF-β), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). Molecular responses were assessed after exposure of cells from the IVD specimens to these cytokines via real-time polymerase chain reaction and immunofluorescence staining.

Results

mRNA gene expression was significantly greater for aggrecan, type I collagen, type II collagen, alkaline phosphatase, osteocalcin, and Sox9 in group 1 than mRNA gene expression in group 2, when the samples were not treated with cytokines. Analysis of mRNA levels for these molecules after morphogen treatment revealed significant increases in both groups, which were much higher in group 1 than in group 2. The average number of IVD cells that were immunofluorescence stained positive for alkaline phosphatase increased after treatment with rhBMP-2 and TGF-β in group 1.

Conclusion

The biologic responsiveness to treatment of rhBMP-2, TGF-β, TNF-α, and IL-1β in the degenerative living human IVD can be different according to the degree of degeneration of the IVD.     

SAHA对糖尿病大鼠肾组织Twist和HDAC1表达的影响

目的:通过观察辛二酰苯胺异羟肟酸(SAHA)对糖尿病(DM)大鼠肾组织Twist、组蛋白去乙酰化酶1(HDAC1)及相关上皮-间充质转化和纤维化指标表达的影响,初步探讨SAHA对糖尿病肾病Twist表达变化的可能干预机制。方法:以链脲佐菌素复制DM大鼠模型,实验分为正常对照(NC)组、DM组和SAHA组,每组12只。造模成功8周后,SAHA组用SAHA(25 mg·kg~(-1)·d~(-1))进行灌胃。治疗8周后处死大鼠,HE和Masson染色观察肾组织形态变化;免疫组织化学染色观察肾组织中Twist的表达变化;Western blot法检测肾组织中Twist、HDAC1、上皮钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)和IV型胶原(Col-Ⅳ)蛋白的表达;real-time PCR检测肾组织中Twist的mRNA表达。结果:DM组的血糖、24 h尿蛋白量和肾脏指数均显著高于NC组;而SAHA组肾脏指数与DM组相比有明显降低(P0.05),24 h尿蛋白量虽有降低,但差异无统计学显著性,而血糖无明显改变。与NC组对比,DM组大鼠肾组织中HDAC1、α-SMA和Col-Ⅳ蛋白表达上调,E-cadherin蛋白表达下调,Twist的mRNA和蛋白表达上调(P0.05);SAHA组大鼠肾组织中的Twist、HDAC1、α-SMA和Col-Ⅳ蛋白表达量明显低于DM组(P0.05),且Twist的mRNA的表达比DM组低,E-cadherin蛋白表达较DM组有所上升(P0.05)。相关性结果显示,在糖尿病大鼠肾组织中,Twist与HDAC1蛋白表达呈正相关(P0.05)。结论:SAHA可下调DM大鼠肾组织中Twist的表达,减轻糖尿病肾病大鼠肾纤维化病变,其机制可能与抑制HDAC1和Twist形成进而促进Ecadherin转录有关。     

Astragaloside effect on TGF-β1, SMAD2/3, and α-SMA expression in the kidney tissues of diabetic KKAy mice

Numerous cytokines participate in the occurrence and development of inflammation and renal interstitial fibrosis. Previous studies confirmed that TGF-β1 overexpressed in diabetic nephropathy. As a downstream signal protein of TGF-β1 family, SMAD has an important role in the process of α-SMA mediated renal interstitial fibrosis. This study aimed to study astragaloside effect on TGF-β1, SMAD2/3, and α-SMA expression in the kidney tissue of diabetic KKAy mice, to reveal its potential impact on renal interstitial fibrosis. 20 type II diabetic KKAy mice were randomly equally divided into model group and astragaloside group, while 10 male C57BL/6J mice were selected as the control. Astragaloside at 40 mg/(kg•d) was given when the KKAy mice fed with high-fat diet to 14 weeks old. The mice were killed at 24 weeks old and the kidney tissue samples were collected. Pathology morphological changes were observed. TGF-β1, SMAD2/3, and α-SMA expression levels were determined by immunohistochemistry. Compared with control, mice kidney in model group appeared obvious fibrosis and up-regulated blood glucose level, TGF-β1, SMAD2/3, and α-SMA expression (P < 0.05). Mice in astragaloside group exhibited alleviated renal interstitial fibrosis compared with the model. Its blood glucose level, TGF-β1, SMAD2/3, and α-SMA expression levels were significantly lower than the model group (P < 0.05). Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1, SMAD2/3, and α-SMA expression.     

Contraction of tubulointerstitial fibrosis tissue in diabetic nephropathy,as demonstrated in an in vitro fibrosis model

Tubulointerstitial fibrosis in diabetic nephropathy (DN) was investigated using an in vitro tissue model of remodeling, to determine the pathogenic mechanism of fibrosis that leads to renal atrophy, i.e., renal failure. The remodeling model consisted of a renal fibroblast-populated collagen lattice (FPCL). The overexpression of transforming growth factor (TGF)-β1 in the diabetic kidney gave rise to FPCL contraction. FPCL relaxation was induced by the subsequent addition of cytochalasin D. The FPCL failed to contract when exposed to TGF-β1 plus Y27632, a Rho kinase inhibitor. TGF-β1 induced the phosphorylation of myosin light chains, and Y27632 blocked this activity. TGF-β1-induced FPCL contraction was suppressed by the addition of 2,3-butanedione monoxime, a myosin ATPase inhibitor. As shown in the video, the contraction rate of the projections of the cells in the FPCL was significantly greater in the TGF-β1 group than in the control group. Collectively, these results indicate that TGF-β1-induced FPCL contraction is attributable to actin–myosin interactions in the fibroblasts through the activation of Rho kinase, the phosphorylation of myosin light chains, and the subsequent activation of myosin ATPase. We propose that via these mechanisms, tubulointerstitial fibrosis generates tissue contraction that leads to renal atrophy and renal failure in DN.     

The effects of palm vitamin E on stress hormone levels and gastric lesions in stress-induced rats

Introduction

This study examines the effects of palm vitamin E (PVE) or α-tocopherol (α-TF) supplementation on adrenocorticotropin hormone (ACTH), corticosterone and gastric lesions in rats exposed to water-immersion restraint stress (WIRS).

Material and methods

Sixty male Sprague-Dawley rats (200-250 g) were divided into three groups. Group I: 20 rats as a control group were given a normal diet. Group II: 20 rats received oral supplementation of PVE at 60 mg/kg body weight. Group III: 20 rats received oral supplementation of α-TF at 60 mg/kg body weight. After the treatment period of 28 days, each group was further subdivided into two groups: 10 rats not exposed to stress, and the other 10 rats subjected to WIRS for 3.5 h. Blood samples were taken to measure the ACTH and corticosterone levels. The rats were then sacrificed and the stomach excised and opened along the greater curvature and examined for lesions.

Results

Rats exposed to WIRS had lesions in their stomach mucosa. Our findings showed that dietary supplementation of PVE or α-TF was able to reduce gastric lesions significantly in comparison to the stressed controls. The WIRS increased plasma ACTH and corticosterone significantly. Palm vitamin E and α-TF treatments reduced these parameters significantly compared to the stressed controls.

Conclusions

Supplementation with either PVE or α-TF reduces the formation of gastric lesions, probably by inhibiting the elevation of ACTH and corticosterone levels induced by stress.     

TGF-β1 gene-modified,immature dendritic cells delay the development of inflammatory bowel disease by inducing CD4+Foxp3+ regulatory T cells

Inflammatory bowel disease (IBD) is caused by an uncontrolled immune response in the intestinal lumen, leading to inflammation in genetically predisposed individuals. Immunotherapy may be a promising approach to the treatment of IBD. Here, we show that transforming growth factor-β1 (TGF-β1) gene-modified immature dendritic cells (imDCs) could enhance the inhibitory function of imDCs and delay the progress of IBD induced by dextran sodium sulfate in mice. The results of fluorescence-activated cell sorter (FACS) demonstrated that this protective effect is mediated partially by inducing CD4⁺Foxp3⁺ regulatory T cells (Tregs) in mesentery lymph nodes to control inflammation. In vitro experiments also supported this hypothesis. In conclusion, we provide evidence that TGF-β1-modified bone marrow-derived imDCs may have a therapeutic effect to IBD.     

Increased IL-12 and decreased IL-33 serum levels are associated with increased Th1 and suppressed Th2 cytokine profile in patients with diabetic nephropathy (CURES-134)

The role played by recently discovered novel cytokine IL-33 in controlling T-helper (Th)1 and Th2 cytokines under conditions of diabetic nephropathy (DN) is less well studied. In the present study, we estimated the levels of IL-33 along with both Th1 and Th2 cytokines in the serum of normal glucose tolerant (NGT), diabetic subjects with (DN) or without nephropathy (DM) and correlated it with the clinical risk factors of diabetes and nephropathy. 222 study subjects were recruited from the Chennai Urban Rural Epidemiology Study (CURES): 61 NGT, 79 DM and 82 DN. IL-33 level was estimated by ELISA while other Th1 (IL-12, IFN-gamma and IL-2) and Th2 (IL-4, IL-5 and IL-13) cytokines were measured using a Bio-plex bead assay. DM subjects showed a mixed Th1-Th2 profile (increased IFN-g, IL-12, IL-4 and IL-13 and decreased IL-33) while DN subjects showed enhanced Th1 profile (increased IFN-g, IL-2 and IL-12) with suppression of Th2 cytokine (decreased IL-33 and IL-13). The IL-33 levels showed a serial decline with increasing severity of insulin resistance and microalbuminuria. DN was associated with enhanced Th1 response and suppression of Th2 responses which might be due to inreased levels of IL-12 and decreased levels of IL-33 cytokines respectively.     

Association of vitamin D receptor gene polymorphisms with susceptibility to asthma in Tunisian children: A case control study

Background

Vitamin D and its nuclear receptor (VDR) are linked to asthma in a genetic and immunologic basis. Polymorphisms in the VDR gene may alter the actions of vitamin D and then influence the development and the severity of asthma.

Aims

We aimed at elucidating the genetic association of VDR gene polymorphisms with susceptibility to asthma in Tunisian children and with serum vitamin D levels.

Methods

The study included 155 patients recruited from Abderrahmen MAMI hospital in Tunisia and two hundred twenty five healthy individuals matched with patients in age and sex for comparison. VDR genotypes were determined by PCR-RFLP method using endonuclease FokI, BsmI, TaqI and ApaI and vitamin D was assessed with a radioimmunoassay kit.

Results

The distribution of genotype frequencies differed significantly between asthmatics and controls (FokI: P = 0.04; BsmI: P = 0.006; TaqI: P = 0.006). Haplotype analyses revealed a significant association between bAt and bat haplotypes and asthma (P = 0.00076, P = 0.016). When patients were stratified according to atopic status and stage of severity, no significant association was detected with VDR variants. No association was found between VDR SNPs and serum 25-hydroxyvitamin D levels.

Conclusion

Our study shows a relation between VDR gene polymorphisms and susceptibility to asthma in children.     

Reverse correlation of Jab1 and Smad4 in PANC-1 cells involved in the pathogenesis of pancreatic cancer

Objective: Steps in the genetic basis of pancreatic cancer (PC) have been recently identified, however, Studies focusing on the relationship between Jab1 and Smad4 in PC are rarely reported. This study was performed to examine the expression patterns and association of Jab1 and Smad4 in PC cells for gaining a further understanding of PC pathogenesis. Methods: Human pancreatic cancer cell line PANC-1 cells were infected with retrovirus vector containing GFP, HA-Jab1, siGFP, and siJab1 respectively. The expression of Jab1 and Smad4 in PANC-1 cells was analyzed by Western blot and immunocytochemistry. Subsequently, the effect of overexpression of Jab1 on cell proliferation inhibition mediated by TGF-β was examined with MTT colorimetry. Results: The expression of Smad4 in PANC-1 cells was inhibited after the overexpression of Jab1. Inversely, the expression of Smad4 was increased after the down-regulation of Jab1 silenced by SiRNA. Smad4 expression in PANC-1 cells was negatively correlated with Jab1 expression. In addition, the cell proliferation inhibitory effect induced by TGF-β in PANC-1 cells was attenuated after the overexpression of Jab1. Conclusions: The reverse correlation of Jab1 and Smad4 in PANC-1 cells may be involved in the Pathogenesis of PC. Jab1 can cause degradation of Smad4 via TGF-β signal pathway, consequently contributing to the proliferation of PC cells.