Role of nestin in the maintenance of the structure of podocytes

AIM:To investigate the expression of nestin, a kind of cytoskeletal protein in cultured murine podocytes and the role of nestin in the maintenance of the podocyte structure.METHODS:The immortalized murine podocytes were cultured. The expression of nestin was determined by immunofluorescence. In differentiated podocytes, the expression of nestin was knock-down by RNAi. The effect of nestin knock-down was examined by Western blotting and immunofluorescence. The projections longer than maximal length of the cell body in which the cells transfected with siRNA and control vector that contained nonhomologous oligo were counted, respectively. RESULTS:Nestin siRNA markedly reduced or abolished nestin expression. In cells transfected with nestin siRNA, the percentage of cells with processes was significantly lower than that in cells transfected with control vector (77.0%±6.3% vs 16.0%±4.6%, n=3, P<0.01). CONCLUSION:Nestin may play an important role in maintaining normal function of podocytes.     

Effect of atorvastatin on neointimal hyperplasia of venous autografts in rats

AIM: To investigate the effect of atorvastatin on neointimal hyperplasia of autogenous vein graft in rats. METHODS: The model of autogenous vein graft was prepared by transplanting the external jugular vein into aorta in Wistar rats. The rats were divided into three groups: sham operation group, graft control group and graft experimental group. From three days after transplantation, the rats of autograft experimental group were treated by atorvastatin at a dosage of 5 mg·kg-1·d-1. Four weeks after treatment, venous autografts were removed at autopsy and cut into 4 μm sections. Histopathological examination was carried out to analysis the neointimal hyperplasia of grafted veins. Immunohistochemical staining was conducted to evaluate SMα-actin and PCNA expression of neointimal cells in venous autografts. RESULTS: In venous autograft control and experimental groups, SMα-actin-positive smooth muscle cells were proliferated and accumulated excessively in venous autografts, which resulted in significant neointimal formation and vascular lumen narrowing. Neointima quantitative assay revealed that the neointimal hyperplasia of venous autografts was suppressed obviously in graft experimental group, and its neointimal area and NIA/MA ratio of venous autografts were significantly lower than those in graft control group (P<0.01). Immunohistochemical assay indicated that the PCNA labeling index of neointimal cells was significantly lower in graft experimental group than that in graft control group (P<0.01). CONCLUSION: Atorvastatin significantly inhibits the proliferation of neointimal smooth muscle cells and the development of neointimal hyperplasia of venous autografts in rats. Atorvastatin is a powerful inhibitor of restenosis after vascular reconstructive operation with a potential for therapeutic use.     

In vitro differentiation of exocrine pancreatic cells from mouse embryonic stem cells

AIM: To explore the effects of sodium butyrate, activin A and dexamthasone on inducing mouse embryonic stem (ES) cells to differentiate into exocrine pancreatic cells in vitro. METHODS: E14 mouse ES cells were cultured in suspension to form embryonic bodies (EBs). The EBs were cultured with differentiating medium containing different concentrations of sodium butyrate, and the spontaneously differentiated ES cells were used as control. Exocrine pancreatic genes such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR at different time points to determine the optimal concentration and exposure time of sodium butyrate. Furthermore, activin A or dexamthasone was also used to explore the effects on exocrine differentiation. After that, the combination of sodium butyrate, activin A and dexamthasone was used to promote the differentiation of exocrine pancreatic cells from ES cells. During the differentiation course, the gene expressions of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected by RT-PCR. Morphological changes were investigated by phase contrast microscopy. Amylase expression was examined by immunofluorescence staining. RESULTS: Exocrine pancreatic gene expressions such as amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase were detected in spontaneously differentiated EBs. A relatively lower concentration of sodium butyrate with a shorter exposure time significantly promoted those above gene expressions as compared to that of spontaneously differentiated EBs. Activin A and dexamethasone induced upregulation of exocrine gene expression. The combination of activin A, sodium butyrate and dexamethasone significantly enhanced the mRNA levels of amylase, chymotrypsinogen, elastase 1, elastase 2 and carboxypeptidase. Under the treatment of activin A, sodium butyrate and dexamethasone, differentiated cells were polygonal in shape with large, round, and center-situated nuclei. According to the observation of immunofluorescence staining, amylase was positive expressed at the final stage. CONCLUSION: These data indicate that exocrine pancreatic differentiation of ES cells is induced by sodium butyrate, activin A and dexamethasone. The combination of pancreatic inducing factors improves the differentiating efficiency.     

Identification of serum differential proteins in colorectal adenoma and early malignantly transformed adenoma by proteomics

AIM: To investigate the associated proteins and sensitive biomarkers for early diagnosis of colorectal adenocarcinoma by comparing the results of differential proteomic analysis between colorectal adenoma and early malignantly transformed adenoma. METHODS: Two-dimensional gel electrophoresis was used to define patterns of protein expressions of colorectal adenoma and early malignantly transformed adenoma. Proteins expressed differentially among groups were detected, cut out and analyzed by MALDI-TOF/TOF mass spectrometry. RESULTS: Two-dimensional protein maps of colorectal adenoma and early malignantly transformed adenoma were analyzed with gel-analysis software, an average of 1 672 spots in adenoma, 1 732 in early malignantly transformed adenoma were observed. 28 spots of a 1.5-fold change were found, including 15 proteins down-regulated and 13 up-regulated in early malignantly transformed adenoma, in which 23 proteins were identified by mass spectrometry, the rate of identification was 82.14%. 13 differential proteins were attained, 8 were up-regulated and 5 were down-regulated, which was classified to 6 categories, including protease inhibitor, complement, immunoglobulin, keratoproteins, signal transduction protein and function-unknown proteins. CONCLUSION: The changes of serum proteins in early malignantly transformed adenoma from adenoma can be identified by proteomic technology. Proteins detected in the study may provide new biomarkers correlated with biological behavior of colorectal adenocarcinoma.     

Effect of simvastatin on expression of integrin-linked kinase and renal tubulointerstitium injury in rats induced by high fat diet

AIM: To explore the effect of simvastatin on expression of integrin-linked kinase (ILK) and the injury of renal tubulointerstitium in the rats induced by high fat diet. METHODS: Fifty-four 6-8 week-old female Wistar rats were randomly assigned to the following three groups: high fat diet group, simvastatin group (rats were fed with high fat diet plus 10 mg·kg-1·d-1 simvastatin) and control group. Six rats in each group were sacrificed at 4th week, 10th week, and the others at 20th week. The injury of renal tubulointerstitium was observed under microscope with HE staining and the expression of renal ILK was determined by Western blotting analysis and immunohistochemistry. Levels of total cholesterol (TC) and triglyceride (TG) in serum were measured by enzymatic colormetric methods. RESULTS: The serum TC and TG levels and the expression of renal ILK significantly increased in both high-fat diet group and simvastatin group, compared to control group at 4th week, reaching a maximum at 20th week (P<0.01). Tubulointerstitium injuries including vacuolar degeneration, syncytial change, clody swelling, necrosis and atrophy of renal tubular epithelial cells, thinning of tubal wall, lumens compensational expansion or even abolition, and inflammatory cell infiltration, interstitial fibrosis were found in both high fat diet group and simvastatin group, compared to control group at 4th week, worsened to a maximum at 20th week. However, all of these ameliorated in simvastatin group, compared to high fat diet group at each time point. CONCLUSION: The results indicate that high-fat diet induces significant lesion of renal tubulointerstitium and increased expression of renal ILK. Simvastatin may play an important role in protecting against tubulointerstitium injury induced by hyperlipoidemia by down-regulating the expression of renal ILK.     

冀西北坝上地区几种蚜虫优势种对菜豆的侵害性研究

采用寄主转换方法研究了冀西北坝上地区几种蚜虫优势种对菜豆的适应性。结果表明,桃蚜、瓜蚜（棉蚜）和菊姬长管蚜转接到露地菜豆 的花、嫩尖、嫩叶等部位上仅能存活6~8 d,不能在菜豆上建立种群。室内寄主转换试验表明,取食菜豆嫩荚的蚜虫存活率明显低于取食茎部嫩尖的 蚜虫存活率。     

大白菜叶片长宽性状的QTL定位

利用一套大白菜双单倍体（DH）群体和包含287个分子标记的遗传图谱,运用复合区间作图法对叶长、无柄叶长、有柄叶长、叶宽、叶柄长、叶柄宽6个叶部形态学性状进行QTL定位及遗传效应的分析。结果表明：在4个连锁群上共检测到12个QTL,分别为叶长相关的3个,无柄叶长相关的2个,有柄叶长相关的2个,叶宽相关的1个,叶柄长相关的2个,叶柄宽相关的2个。各位点的遗传贡献率介于7.5 %~18.9 %之间。其中,叶长与叶柄长的QTL存在共定位,叶宽与叶柄宽的QTL也存在共定位。     

叶面喷施氨基酸水溶肥料对大白菜产量和VC含量的影响

以北京新3号大白菜为试验材料,在大白菜包心期喷施不同浓度氨基酸水溶肥料,测定大白菜产量及VC含量。结果表明：叶面喷施含氨基酸水溶肥料,大白菜产量提高5.8 %~12.4 %、VC含量增加15.8 %~51.7 %。     

钾素不同用量对莴苣硝酸盐积累的影响

以莴苣为试材,采用通用随机排列设计,利用基质培育方法,研究不同供应水平的钾素营养对莴苣硝酸盐积累的影响.结果表明:在供试条件下,硝酸盐含量与硝酸还原酶活性在一定浓度范围内都随着钾素浓度的增大而降低.     

不周土壤水分条件下萝卜幼苗叶片生理特性研究

在不同的土壤含水量条件下,测定了萝卜幼苗叶片的水分状况、叶绿素含量、光合速率、蒸腾速率及呼吸强度等指标.结果表明:随着土壤含水量的降低,萝卜幼苗叶片的相对含水量和水势下降,而且离体叶片的保水力在自然气干时下降较快;叶片叶绿素含量、光合速率、蒸腾速率和呼吸强度也随着土壤含水量的降低都减小.土壤含水量在20%～25%时,萝卜幼苗对土壤干旱的适应性最强.