Role of ILK siRNA on expression of GSK-3β and β-catenin in human tubular epithelial cells stimulated by high glucose

AIM：To investigate the effects of siRNA targeting integrin-linked kinase (ILK) on the expression of glycogen synthase kinase 3β (GSK-3β) and β-catenin during epithelial-mesenchymal transition (EMT) in human kidney proximal tubular epithelial cell line HKC induced by high glucose. METHODS：HKC cells were divided into 4 groups:normal glucose (NG) group, high glucose (HG) group, HG+HK (a vector containing the non-specific siRNA designed as negative control) group and HG＋ILK siRNA group. The inverted fluorescence microscope was used to examine the expression of green fluorescent protein (GFP). The expression of ILK at mRNA and protein levels was detected by RT-PCR and Western blotting. The expression of p-GSK-3β and β-catenin was observed by immunocytochemical staining. The protein expression of total GSK-3β, p-GSK-3β, nuclear β-catenin, total β-catenin, E-cadherin and α-smooth muscle actin (α-SMA) was measured by Western blotting. RESULTS：GFP was observed in HKC cells, indicating that the transfection was successful. Both the protein and mRNA of ILK were down-regulated in HG＋ILK siRNA group compared with HG group and HG+HK group, but still higher than those in NG group. Silencing of ILK down-regulated the expression of p-GSK-3β and nuclear β-catenin. No difference of total GSK-3β or total β-catenin was observed among the 4 groups. CONCLUSION:These data support a functional role of ILK, GSK-3β and β-catenin in tubular EMT induced by high glucose. ILK may promote tubular EMT by regulating the activity of GSK-3β and β-catenin, the downstream effectors of the Wnt/β-catenin pathway.     

Inflammatory response in rat kidney with contrast-induced nephropathy

AIM：To observe the expression of tumor necrosis factor α (TNF-α) and nuclear factor κB (NF-κB) in the renal tissue of the rats with contrast-induced nephropathy (CIN). METHODS：Male Sprague-Dawley rats (n=96) were randomly divided into control group (n=48) and CIN group (n=48). The model rats in CIN group were intravenously injected with iodinated contrast media (76% compound diatrizoate injection,10 mL/kg), while the rats in control group were injected with the same volume of saline. Six rats in each group were sacrificed at 6 h, 12 h, 24 h, 48 h, 72 h, 5 d, 10 d and 15 d after intravenous injection, respectively, and the blood and kidney samples of the rats were obtained. The renal tubular injury was assessed by histological examination (HE staining). The expression of kidney injury molecule-1 (KIM-1), TNF-α and NF-κB at mRNA and protein levels in the renal tissues were semiquantitatively measured by the methods of RT-PCR and immunohistochemistry, respectively. The correlations between the expression of TNF-α, NF-κB and tubular injury score, KIM-1 expression in renal tissue of CIN group were analyzed. RESULTS：The levels of serum creatinine (SCr) and blood urea nitrogen (BUN) in control group were not changed between different time points (P>0.05). The levels of SCr and BUN in CIN group displayed significant increases at different time points (except 15 d) compared with control group (P<0.05). The renal tubular injury score in CIN group was significantly higher at all time points than that in control group (P<0.05). The expression of KIM-1, TNF-α and NF-κB at mRNA and protein levels up-regulated significantly at 6 h and the peaking of KIM-1 expression was at 24 h, while the peaking of TNF-α and NF-κB expression was at 48 h in CIN group. The expression of KIM-1,TNF-α and NF-κB was significantly increased in CIN group compared with control group except at 15 d (P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels showed close correlations with renal tubular injury score (r=0.843, 0.758, 0.743 and 0.707, P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels was also positively correlated with KIM-1 expression (r=0.863, 0.807, 0.839 and 0.855, P<0.05). CONCLUSION：The expression of TNF-α and NF-κB at mRNA and protein levels in the renal tissues of CIN group is up-regulated and is closely related with renal tubular injury, indicating that the inflammatory response is involved in the pathogenesis of CIN.     

Aortic expression of HSP22, TNF-α and eNOS in rats with hyperlipidemia and effects of atorvastatin

AIM:To establish a rat hyperlipidemia model for studying the aortic expression of heat shock protein 22 (HSP22), tumor necrosis factor alpha (TNF-α) and endothelial nitric oxide synthase (eNOS) and the effect of atorvastatin intervention. METHODS:Hyperlipidemia model was established in SD rats. Afterwards, the rats were divided into normal control group, high fat group and high fat+atorvastatin intervention group. The expression of HSP22 and TNF-α in the rat aortas was detected by immunohistochemical assay and the expression of eNOS was assessed by Western blotting. RESULTS:No detectable expression of HSP22 and TNF-α in the normal control group was observed. However, the expression of HSP22 and TNF-α was positive in the high fat group and the atorvastatin intervention group. The mean densities of HSP22 and TNF-α positive particles were significant lower in the atorvastatin intervention group as compared with high fat group (both P＜0.05). The expression of eNOS protein in the high fat group and atorvastatin intervention group was significantly lower than that in normal control group (P<0.01). However, no marked difference of eNOS protein expression between high fat group and atorvastatins intervention group was observed. CONCLUSION: The expression of HSP22 and TNF-α in the rat aortas is increased in the hyperlipidemia rat model. This effect can be restored by atorvastatin treatment. The expression of eNOS in the rat aortas is decreased in the hyperlipidemia rat model, but this tendency could not be attenuated by atorvastatin.     

Hydrogen sulfide ameliorates high glucose-induced endothelial cell senescence by suppressing oxidative stress

AIM：To explore the effect of hydrogen sulfide on the senescence of human umbilical vein endothelial cells (HUVECs) induced by high glucose. METHODS：Senescence model was established by treating HUVECs with 33 mmol/L glucose for 48 h. The parameters were detected to demonstrate the effect of hydrogen sulfide on senescence and the mechanism involved was also investigated. RESULTS：In the cells treated with high glucose, the proliferation was attenuated with a higher number of senescence-associated β-galactosidase (SA-β-Gal) positive cells, and plasminogen activator inhibitor 1 (PAI-1) protein expression, malondialdehyde (MDA) production and NF-κB p65 activity were increased significantly, but the expression of superoxide dismutase 1 (SOD1) was decreased. However, the cell number and SOD1 expression were increased, and the number of SA-β-Gal positive cells, PAI-1 protein expression, MDA production and the activity of NF-κB p65 were decreased after sodium hydrosulfide (100 and 200 μmol/L) treatment.CONCLUSION：Exo-genous hydrogen sulfide prevents HUVECs against high glucose-induced senescence by suppressing oxidative stress and NF-κB p65 activity.     

Mechanisms of hyperglycemia induced by immunosuppressant FK506

AIM：To investigate the effect of immunosuppressant FK506 on serum glucose in rats and to explore its mechanism. METHODS：Sprague-Dawley rats (n=12) were randomly divided into drug group and normal group. The rats in drug group were intraperitoneally injected with FK506 at dose of 1 mg·kg-1·d-1 and the rats in normal group received saline (1 mL·kg-1·d-1, ip) for 14 d. The fasting weight and fasting glucose were regularly measured every 2 d. Visceral fat was isolated from the rats at the end of experiment. The mRNA expression of adiponectin, leptin, visfatin, resistin, retinol-binding protein 4 (RBP4) and peroxisome proliferator-activated receptors γ (PPAR-γ) was determined by real-time fluorescence quantitative PCR. The protein expression of PPAR-γ and adiponectin was measured by Western blotting. RESULTS：Compared with normal group, the concentration of fasting blood glucose in model group was significantly increased from the 10th day (P<0.05). At day 14, the fasting blood glucose of the model group increased from (5.10±062) mmol/L to (7.73 ± 0.73) mmol/L. No significant change of blood glucose in normal group between the 10th day and the 14th day ［from (4.66 ± 0.32) mmol/L to (5.80±0.10) mmol/L］ was observed. Compared with normal group, the mRNA expression of PPAR-γ, adiponectin and leptin in the adipose tissue of model group was significantly decreased (P<001), whereas the expression of visfatin, resistin and RBP4 was significantly increased (P<005). Compared with normal group, the expression of PPAR-γ and adiponectin in model group was decreased (P<001). CONCLUSION：FK506 may decrease the expression of PPAR-γ to change the expression of adipocytokines and induce hyperglycemia in rats.     

番茄新品种潍科粉3 号的选育

潍科粉3 号是以H10A 为母本,以J10211 为父本杂交选育而成的粉果番茄一代杂种。早熟,由定植到第1 穗果成
熟需75 d（天）左右,植株长势强,无限生长类型,连续坐果能力强,始花节位在第6～7 节;果实略高圆形,着色均匀,
亮粉色,果实味浓爽口,风味佳,VC 含量高达256 mg·kg-1;硬度高,萼片平展,商品性好,单果质量230～250 g,每
667 m² 产量13 000 kg 以上;对番茄黄化曲叶病毒、疫病、叶霉病的抗性较对照梦之粉强。适于温室和大棚早春、秋延后及
越冬栽培。     

不同时期接种AM 真菌对大棚黄瓜产量及品质的影响

研究不同时期接种丛枝菌根真菌根内球囊菌（Glomus intraradices,GI）对大棚土壤栽培黄瓜植株生长、产量及营
养品质的影响。结果表明：黄瓜不同时期接种GI 对其生长均有促进作用,与不接菌的对照相比,播种时接菌和移栽时接菌处理使黄瓜的单株产量分别提高54%、34%;播种时接菌处理的黄瓜叶片的叶绿素含量比对照显著提高了14.7%～17.7%;播种时接菌和移栽时接菌的黄瓜硝酸盐含量较对照分别下降了9.4% 和15.0%,总糖含量分别比对照提高了8.9% 和7.1%;移栽时接菌处理的黄瓜粗蛋白含量比对照提高了4.6%;播种时接菌和移栽时接菌显著提高了黄瓜VC 含量。表明接种GI 对黄瓜生长和产量品质具有一定的改善作用。     

厚皮甜瓜新品种敦蜜1 号的选育

敦蜜1 号是以自交系ML29 为母本,以自交系ML20 为父本配制而成的厚皮甜瓜一代杂种。中早熟,露地栽培生育
期87 d（天）左右。果实发育期40 d（天）左右。植株生长势强,叶色深绿。果实椭圆形,果形指数1.4,果皮纯白色带网纹,
果肉浅绿色,肉厚3.4 cm,中心可溶性固形物含量16.4%,耐贮运。单瓜质量1.5 kg 左右,每667 m² 产量2 260 kg 左右。抗
甜瓜蔓枯病、霜霉病、白粉病。适宜于酒泉市及类似区域大田地膜覆盖种植。     

马铃薯新品种天薯12 号的选育

天薯12号是以品系天97-8-98为母本,庄薯3号为父本杂交选育而成的马铃薯新品种。从出苗至块茎成熟126 d（ 天）
左右,属晚熟品种。薯块椭圆形,黄皮黄肉,芽眼浅而紫。单株结薯4.1 个,平均单薯质量105 g,大中薯率87.4%。每667 m²
产量在1 500 kg 以上,适于甘肃天水、临夏、定西、平凉、陇南等地种植。     

保护地番茄新品种洛番12 号的选育

洛番12 号是以992 为母本,978 为父本杂交育成的适宜保护地栽培的番茄一代杂种。无限生长类型,中早熟,生长势强,普通花叶,叶色绿,第8～9 节着生第１花序,花序间隔2～3 叶,果实圆形,成熟果粉红色,果面光滑,色泽鲜亮,无绿果肩,果脐及梗洼小,果实硬,整齐度好,单果质量200 g 左右,平均货架期为8 d（天）,可溶性固形物含量4.8%。耐寒、耐弱光,不易裂果,耐贮运。早春保护地栽培每667 m2 产量7 000 kg 左右。