Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells     

Wei Wang  Li Wang  Xin-xiu Li  Xia Chen  Hai-yan Zhang  Yu He  Jing-jing Wang  Yong-yan Zhao  Bao-le Zhang  Yin-xue Xu 《Journal of Zhejiang University. Science. B》2010,11(9):719-727

Bone morphogenetic proteins (BMPs) play a critical role in the growth and steroidogenesis of granulosa cells (GCs). BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors. Upon ligand binding, BMPs activate intracellular Smad proteins and regulate growth and apoptosis in various cell types. The objective of this study was to demonstrate the effects of BMP/Smad signal on growth and steroidogenesis of porcine GCs. A strategy of RNA interference (RNAi)-mediated ‘gene silencing’ of Smad4, a core molecule mediating the intracellular BMP/Smad signal transduction pathways, was used to interrupt endogenous BMP/Smad signaling. Results indicate that Smad4-small interfering RNA (siRNA) caused specific inhibition of Smad4 mRNA and protein expression after transfection. Interrupted endogenous BMP/Smad signaling significantly inhibited growth, and induced apoptosis of porcine GCs, while decreasing estradiol production. In addition, interrupted BMP/Smad signaling significantly (P<0.05) changed the expression of Cyclin D2, CDK4, Bcl-2, and Cyp19a1. These findings provide new insights into how BMP/Smad signaling regulates the growth and steroidogenesis of porcine GCs.  相似文献   

Effect of high glucose levels on the calcification of vascular smooth muscle cells by inducing osteoblastic differentiation and intracellular calcium deposition via BMP-2/Cbfα-1 pathway     

Liu F  Zhong H  Liang JY  Fu P  Luo ZJ  Zhou L  Gou R  Huang J 《Journal of Zhejiang University. Science. B》2010,11(12):905-911

In this paper,we investigate the effect and the possible mechanism of high glucose levels on the calcification of human aortic smooth muscle cells (HASMCs).HASMCs were divided into four groups: normal glucose group (NG),osmolality control group (OC),high glucose group (HG,HASMCs culture medium containing 30 mmol/L glucose),and high glucose plus recombinant human Noggin protein (bone morphogenetic protein-2 (BMP-2) antagonist) group (HN).The mRNA levels and the protein expressions of BMP-2 and core binding factor alpha-1 (Cbfα-1) were measured by real-time quantitative polymerase chain reaction (PCR) and Western blot.After induced by 10 mmol/L β-glycerol phosphoric acid,cells were harvested for assessments of alkaline phosphatase (ALP) activities at Days 1,2,and 3,and intracellular calcium contents at Days 7 and 14,respectively.High glucose levels increased the mRNA levels and the protein expressions of BMP-2 and Cbfα-1 (P0.05).The expression of Cbfα-1 was partially blocked by Noggin protein (P0.05),while BMP-2 was not (P0.05).After being induced by β-glycerol phosphoric acid,high glucose levels increased the ALP activity [(48.63±1.03) vs.(41.42±2.28) U/mg protein,Day 3;P0.05] and the intracellular calcium content [(2.76±0.09) vs.(1.75±0.07) μmol/mg protein,Day 14;P0.05] in a time-dependent manner when compared with the NG group,while the ALP activity could not be blocked by Noggin protein [(48.63±1.03) vs.(47.37±0.97) U/mg protein,Day 3;P0.05].These results show that high glucose levels can evoke the calcification of HASMCs by inducing osteoblastic trans-differentiation and intracellular calcium deposition via the BMP-2/Cbfα-1 pathway,which can be partially blocked by Noggin protein.  相似文献   

Clinicopathological features of atypical lipomatous tumors of the laryngopharynx     

Shi HY  Wei LX  Wang HT  Sun L 《Journal of Zhejiang University. Science. B》2010,11(12):918-922

Atypical lipomatous tumor (ALT) of the laryngopharynx is rare. Here we report five cases to demonstrate their clinicopathological features. The patients were four males and one female, aged 41 to 69 years (median 53.6 years). All tumors (two in the hypopharynx and three in the larynx) presented as a slowly growing, painless mass. Symptoms included dysphagia (2/5), dysphonia (3/5), and the feeling of a foreign body in the throat (5/5). Tumors were well circumscribed or focally infiltrative, ranging from 2.0 to 5.0 cm (median, 3.4 cm) in size, and microscopically showed the typical features of lipoma-like ALT. Immunohistochemically, tumor cells were stained with S-100, vimentin, murine double minute 2 (MDM-2), and cyclin-dependent kinase 4 (CDK4). Two patients had local tumor recurrences at 6 and 14 months after initial surgery during follow-up. ALT of laryngopharynx is an indolent tumor. Immunohistochemical staining for MDM-2 and CDK4 is helpful in pathological diagnosis.  相似文献   

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk   总被引：2，自引：0，他引：2  

Bin Huang  Yun Yin  Lei Lu  Hai Ding  Lin Wang  Ting Yu  Jia-jin Zhu  Xiao-dong Zheng  Yan-zhen Zhang 《Journal of Zhejiang University. Science. B》2010,11(10):812-818

A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC₅₀) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%–84.60%, with coefficients of variation of less than 12.2%.  相似文献   

Case Report :Surgical intervention of severe post-ERCP-pancreatitis accompanied with duodenum perforation     

Zuo-bing CHEN  Zhong-yan LIANG  Yun ZHANG  Shao-yang ZHANG  Shu-sen ZHENG 《Journal of Zhejiang University. Science. B》2010,11(1)

Endoscopic retrograde cholangiopancreatography (ERCP) is a procedure widely used to diagnose and treat conditions of biliary or pancreatic ductal system. The post-ERCP severe acute pancreatitis (SAP) accompanied with duodenum perforation is rare but serious, remaining a challenge in clinic. In this study we report two such cases. Two Chinese women were treated for clinical suspicion of bile duct obstruction and underwent ERCP after admission. Both developed duodenum perforation and SAP after ERCP, and were ...  相似文献   

Arachnoid cell involvement in the mechanism of coagulation-initiated inflammation in the subarachnoid space after subarachnoid hemorrhage     

Zhao-liang Xin  Xiao-kang Wu  Jian-rong Xu  Xi Li 《Journal of Zhejiang University. Science. B》2010,11(7):516-523

Objective: To assess if arachnoid cells have the capability to present antigen and activate T-lymphocytes after stimulation by bloody cerebrospinal fluid (CSF), and to illuminate the mechanism of coagulation-initiated inflammation in the subarachnoid space after subarachnoid hemorrhage (SAH). Methods: Arachnoid cells were cultured, characterized, and examined by immunofluorescence for the basal expression of human leukocyte antigen-DR (HLA-DR). Expression of HLA-DR, after co-culturing arachnoid cells in vitro with bloody CSF, was investigated by immunofluorescence and flow cytometry (FCM). The variation of arachnoid cells' ultrastructure was observed by transmission electron microscope (TEM). Arachnoid cells were co-cultured with peripheral blood mononuclear cells (PBMCs). The content of soluble interleukin-2 receptor (sIL-2r) in culture medium was detected by enzyme-linked immunosorbent assay (ELISA). Results: (1) Arachnoid cells were successfully cultured for many passages. The immunofluorescent staining was positive for HLA-DR in over 95% of the human arachnoid cells. The punctate HLA-DR was distributed in cytoplasm and not in the karyon. (2) After co-culturing arachnoid cells in vitro with bloody CSF, numerous particles with strong fluorescence appeared in the cytoplasm on Day 6. On Day 8, the quantity of particles and fluorescent intensity were maximal. FCM showed that the percentage of HLA-DR expressing cells was (2.5±0.4)% at the first 5 d, increasing to (60.8±3.6)% on Day 7. (3) After co-culturing arachnoid cells in vitro with bloody CSF, many lysosome and secondary lysosome particles were present in the cytoplasm. Hyperplasia of rough endoplasmic reticulum and enlarged cysts were observed, with numerous phagocytizing vesicles also observed at the edge of the arachnoid cells. (4) Arachnoid cells stimulated by bloody CSF were co-cultured in vitro with PBMCs. The content of sIL-2r in the culture medium, having been maintained at around 1.30 ng/ml during the first 3 d, had increased by Day 4. The content of sIL-2r peaked 7.53 ng/ml on Day 7 and then reduced gradually. Conclusions: (1) Basic HLA-DR expression is present in arachnoid cells. (2) After stimulation by bloody CSF, arachnoid cells have the potential to serve as antigen presenting cells (APCs) and the ability to activate T-lymphocytes, indicating that arachnoid cells are involved in the mechanism of coagulation-initiated inflammation in the subarachnoid space after SAH.  相似文献   

Validity and reliability testing of the Chinese (mainland) version of the 39-item Parkinson’s Disease Questionnaire (PDQ-39)     

Wei Luo  Xiao-hong Gui  Bo Wang  Wen-ying Zhang  Zhi-yuan Ouyang  Yi Guo  Bao-rong Zhang  Mei-ping Ding 《Journal of Zhejiang University. Science. B》2010,11(7):531-538

The 39-item Parkinson’s Disease Questionnaire (PDQ-39) has been tested in many languages, but not in Chinese mainland. We aimed to assess the Chinese (mainland) version of the PDQ-39. Seventy-one subjects with Parkinson’s disease (PD) completed the PDQ-39 and the Medical Outcomes Study 36-item Short Form Health Survey (SF-36). All subjects were retested with the PDQ-39 a week later. The united Parkinson’s disease rating scale (UPDRS) and the Hoehn and Yahr (H &; Y) scale were also used to evaluate the subjects. Reliability was assessed by Cronbach’s α and intra-class correlation coefficient (ICC). Validity was examined in terms of agreement with SF-36, UPDRS, and H &; Y scales. The Chinese (mainland) version of the PDQ-39 demonstrated acceptable reliability (Cronbach’s α: 0.84–0.88; ICC: 0.56–0.82). The item-total correlations (0.33–0.88) and scaling success rates (77.56%) indicated satisfactory convergent and discriminant validity of the PDQ-39 items. The correlations between related constructs of the PDQ-39 and UPDRS (r=0.44–0.68) and between those of the PDQ-39 and SF-36 (r=(−0.46)-(−0.69)) were all statistically significant (P<0.01). Except for stigma, cognitions, and bodily discomfort, all other dimensions of the PDQ-39 significantly discriminated patients at different H &; Y stages indicated by the H &; Y scale. Although our observations indicate that some problematic subscales of this version of the PDQ-39 could be improved upon, this study suggests acceptable reliability and validity of the Chinese (mainland) version of the PDQ-39.  相似文献   

Transgenic barley with overexpressed PTrx increases aluminum resistance in roots during germination     

Li QY  Niu HB  Yin J  Shao HB  Niu JS  Ren JP  Li YC  Wang X 《Journal of Zhejiang University. Science. B》2010,11(11):862-870

A transgenic barley line (LSY-11-1-1) with overexpressed Phalaris coerulescens thioredoxin gene (PTrx) was employed to measure the growth, protein oxidation, cell viability, and antioxidase activity in barley roots during germination on the presence of 2 mmol/L AlCl₃ on filter paper. The results show that (1) compared with the non-transgenic barley, LSY-11-1-1 had enhanced root growth, although both were seriously inhibited after AlCl₃ treatment; (2) the degree of protein oxidation and loss of cell viability in roots of LSY-11-1-1 were much less than those in roots of non-transgenic barley, as reflected by lower contents of protein carbonyl and Evans blue uptakes in LSY-11-1-1; (3) activities of catalase (CAT), glutathione peroxidase (GPX), ascorbate peroxidase (APX), and glutathione reductase (GR) in LSY-11-1-1 root tips were generally higher than those in non-transgenic barley root tips, although these antioxidase activities gave a rise to different degrees in both LSY-11-1-1 and non-transgenic barley under aluminum stress. These results indicate that overexpressing PTrx could efficiently protect barley roots from oxidative injury by increasing antioxidase activity, thereby quenching ROS caused by AlCl₃ during germination. These properties raise the possibility that transgenic barley with overexpressed PTrx may be used to reduce the aluminum toxicity in acid soils.  相似文献   

A facile approach to construct hybrid multi-shell calcium phosphate gene particles     

Zhi-xue Xu  Ran Zhang  You-xiang Wang  Qiao-ling Hu 《Journal of Zhejiang University. Science. B》2010,11(4):292-297

The calcium phosphate (CaP) particles have attracted much attention in gene therapy. How to construct stable gene particles was the determining factor. In this study,hybrid multi-shell CaP gene particles were successfully constructed. First,CaP nanoparticles served as a core and were coated with DNA for colloidal stabilization. The ξ-potential of DNA-coated CaP nanoparticles was -15 mV. Then polyethylenimine (PEI) was added and adsorbed outside of the DNA layer due to the electrostatic attraction. The ξ-pot...  相似文献   

(1+1+1)维时空光孤子控制     

郝瑞宇  琚爱堂 《长治学院学报》2010,27(2):1-3

本文通过数值地求解孤子控制系统下的（1＋1＋1）维非线性薛定谔方程,讨论在一定的孤子控制系统中传输的时空光孤子。结果表明,在一定的孤子控制系统中,（1＋1＋1）维时空光孤子可较稳定地传输。最后,讨论时空光孤子传输过程的稳定性。结果表明,在白噪声扰动下,时空光孤子传输过程是稳定的。  相似文献