ASSOCIATION BETWEEN LOW- DENSITY LIPOPROTEIN RECEPTOR- RELATED PROTEIN GENE, BUTYRYLCHOLINESTERASE GENE AND ALZHEIMER'S DISEASE IN CHINESE

Objective. To research the relations between low- density lipoprotein receptor- related protein gene (LRP) polymorphism, butyrylcholinesterase gene (BchE) polymorphism and Alzheimer’s disease (AD) in Chinese. Methods. The gene polymorphisms of LRP and BchE were genotyped in 38 AD cases and 40 controls with polymerase chain reaction- restriction fragment length polymorphism (PCR- RFLP) methods. AD groups were classified according to the LRP C/C genotype and compared with matched controls. Results. AD group had higher frequencies of C/C homozygote (81.6％ vs 60.0％ , P< 0.05) and of C allele (89.5％ vs 76.3％ , P< 0.05),with no significant difference between any of these LRP genotypes classified AD groups and their respective control groups. Conclusions. A positive correlation was found between LRP gene polymorphism and AD, but not between BchE gene polymorphism and AD in Chinese AD cases.

Interaction between insulin-like growth factor binding protein-related protein 1 and transforming growth factor beta 1 in primary hepatic stellate cells

BACKGROUND: We previously showed that insulin-like growth factor binding protein-related protein 1 (IGFBPrP1) is a novel mediator in liver fibrosis. Transforming growth factor beta 1 (TGF beta 1) is known as the strongest effector of liver fibrosis. Therefore, we aimed to investigate the detailed interaction between IGFBPrP1 and TGF beta 1 in primary hepatic stellate cells (HSCs). METHODS: We overexpressed TGF beta 1 or IGFBPrP1 and inhibited TGF beta 1 expression in primary HSCs for 6, 12, 24, 48, 72, and 96 hours to investigate their interaction and observe the accompanying expressions of a-smooth muscle actin (alpha-SMA), collagen I, fibronectin, and phosphorylated-mothers against decapentaplegic homolog 2/3 (p-Smad2/3). RESULTS: We found that the adenovirus vector encoding the TGF beta 1 gene (AdTGF beta 1) induced IGFBPrP1 expression while that of alpha-SMA, collagen I, fibronectin, and TGF beta 1 increased gradually. Concomitantly, AdIGFBPrP1 upregulated TGF beta 1, alpha-SMA, collagen I, fibronectin, and p-Smad2/3 in a time-dependent manner while IGFBPrP1 expression was decreased at 96 hours. Inhibition of TGF beta 1 expression reduced the IGFBPrP1-stimulated expression of alpha-SMA, collagen I, fibronectin, and p-Smad2/3. CONCLUSIONS: These findings for the first time suggest the existence of a possible mutually regulation between IGFBPrP1 and TGF beta 1, which likely accelerates liver fibrosis progression. Furthermore, IGFBPrP1 likely participates in liver fibrosis in a TGF beta 1-depedent manner, and may act as an upstream regulatory factor of TGF beta 1 in the Smad pathway.

Expression levels of autophagy related proteins and their prognostic significance in retinocytoma and retinoblastoma

·AIM: To discuss the prognostic significant of autophagy related proteins(ARPs) in retinoblastoma(RB)and to find the molecular marker to distinguish retinocytoma(RC) and RB by investigating the different expression profiling of microtubule-associated protein light chain 3(LC3B) and other ARPs in RC and RB.·METHODS: Specimens with retinocytoma region(RCR)or mainly composed with Flexner-Winterstein rosettes(FWR) were screen out from 219 paraffin-embedded RB samples and respectively taken as RCR group and FWR group. Others were taken as undifferentiated(UD) group.Immunochemistry(IHC) of LC3 B and electronic microscopy was used to identify autophagy. The IHC scores of LC3 B and other ARPs, such as Beclin, PTEN,p27, p16INK4 a, mTOR and BCL-2 were compared and correlation analysis was applied to find potential proteins which may involve in autophagy regulation. The prognostics significance of LC3 B was evaluated by comparing the high risk features(HRFs) in 3 groups of total 219 samples.·RESULTS: Twenty-one specimens with RCR and 36 specimens mainly composed with FWR were screen out.RCR cell had a high level of LC3 B and lots of autophagic vacuoles. Beclin, PTEN, p27 had positive correlation with LC3, and p16INK4 ahad negative correlation, while the expression of mTOR and BCL-2 in RCR and RB region did not show any difference. Cases with RCR had lower rate of HRFs than undifferentiated cases.·CONCLUSION: ARPs had different expression pattern between RCR and other pathological types of RB, and could be ideal markers to distinguish RC from RB. Our finding indicated cases with RCR had favorable prognosis just like those with FWR.

Autophagy related protein 9A increase in hepatitis B virusassociated hepatocellular carcinoma and the role in apoptosis

The majority of hepatocellular carcinoma(HCC)cases are associated with the hepatitis B virus(HBV)infection.Autophagy related protein 9A(ATG9A)is a transmembrane protein required for autophagosome formation.In order to investigate the role of ATG9A in HBV-associated HCC,ATG9A protein expression was determined in tumor liver tissues and compared with adjacent nontumor tissues from HCC patients with or without HBV infection.In HBVassociated HCC tissues,ATG9A protein level was increased in tumor liver tissues,but not in cases of non-HBV HCC.Our findings suggested that ATG9A might be involved in HBV and cancer cell survival.Therefore,we aimed to analyze the function of ATG9A in HBV replication using RNA interference to evaluate the HBV DNA level using real-time PCR.In the present study,there were no significant differences between shATG9A-transfected HepG2.2.15 cells and the mock control.However,we found that silencing ATG9A affected apoptosis in HepG2.2.15 and HepG2 cell lines.Our results indicated that ATG9A might be partly involved in the survival of HCC.Thus,the inhibition of ATG9A together with other targets might be a potential drug target for HCC treatment.

The Wheat Pathogenesis Related Protein (TdPR1.2) Ensures Contrasting Behaviors to <i>E. coli</i>Transformant Cells under Stress Conditions

The pathogenesis-related proteins 1 (PR-1) gene family play important roles in the plant metabolism in response to biotic and abiotic stresses. The wheat TdPR1.2 has been previously isolated and characterized. Here we showed by bio-informatic analysis that TdPR1.2 contains six cysteine residues that are conserved between all PR-1 proteins tested. Using ScanProsite tool, we found that TdPR1.2 structure has a CRISP family signature 1 and 2 located at the C-terminal part of the protein. Those two domains are conserved in many identified PR1.2 proteins in plants. Moreover, SignalIP-5.0 analysis revealed that TdPR1.2 contains a putative signal peptide formed by 25 amino acids at the N-terminal extremity. The presence of this signal peptide suggested that the mature proteins will be secreted after the cleavage of the signal sequence. Further, we investigate the role of the TdPR1.2 proteins in the growth of Escherichia coli transformants cells under different abiotic stresses. Our results showed that the full-length form of TdPR1.2 enhanced tolerance of E. coli against salt and osmotic stress but not to KCl. Moreover, TdPR1.2 protein confers bacterial tolerance to heavy metals in solid and liquid mediums. Based on these results, we suggest that the TdPR1.2 protein could play an important role in response to abiotic stress conditions.

Down-regulation of lung resistance related protein by RNA interference targeting survivin induces the reversal of chemoresistances in hepatocellular carcinoma

Background Both survivin and lung resistance related protein （LRP） are hepatocellular carcinoma （HCC）. But the relationship between survivin and LRP is investigate the effects of down-regulation of survivin on LRP expressions and the both in vitro and in vivo. related to the chemoresistances in ndefinite. The aim of this study was to reversal of chemoresistances in HCC Methods The expressions of survivin were detected by RT-PCR and Western blotting in HCC cell line SMMC-7721 and SMMC-7721/ADM. The sensitivities of these two cell lines to ADM were evaluated by MTT assays. SiRNA which targeted survivin was transfected into SMMC-7721/ADM cells, then the sensitivity of SMMC-7721/ADM cells to ADM and the expressions of survivin and LRP were detected respectively. SMMC-7721/ADM cells were transplanted subcutaneously into nude mice to establish xenograft tumors. Antitumor activities of RNA interference （RNAi） targeting survivin, various doses of ADM and combination therapies were observed respectively. Possible toxicities were evaluated. LRP expression changes were tested. Student＇s ttest was used for evaluating statistical significance. Results The expressions of survivin in SMMC-7721/ADM cell line showed significant elevation compared to those in SMMC-7721 cell line （P 〈0.05）. Positive siRNA down-regulated the expressions of survivin significantly （P 〈0.05）. SiRNA targeting survivin could sensitize SMMC-7721/ADM cells to ADM and down-regulate the expressions of LRP significantly （P 〈0.05）. Growths of the tumors were significantly inhibited in positive siRNA group as compared with those in the control group from the 8th day （P 〈0.05）. Combination therapies caused significant tumor inhibitions compared with tumors of nude mice in the other three groups respectively （P 〈0.05）. No toxicities were found in nude mice treated by siRNA and combination therapies. The expressions of LRP were markedly reduced in tumors treated with siRNA targeting survivin （P 〈0.05）. Conclusions Down regulation of survivin gene by RNAi can increase chemosensitivity of HCC both in vitro and in vivo. The reversal of drug resistance may be reduced through the inhibitions of LRP.

Aging related methylation influences the gene expression of key control genes in colorectal cancer and adenoma

AIM To analyze colorectal carcinogenesis and age-related DNA methylation alterations of gene sequences associated with epigenetic clock CpG sites. METHODS In silico DNA methylation analysis of 353 epigenetic clock Cp G sites published by Steve Horvath was performed using methylation array data for a set of 123 colonic tissue samples [64 colorectal cancer(CRC), 42 adenoma, 17 normal; GEO accession number: GSE48684]. Among the differentially methylated agerelated genes, secreted frizzled related protein 1(SFRP1) promoter methylation was further investigated in colonic tissue from 8 healthy adults, 19 normal children, 20 adenoma and 8 CRC patients using bisulfite-specific PCR followed by methylation-specific high resolution melting(MS-HRM) analysis. m RNA expression of age-related "epigenetic clock" genes was studied using Affymetrix HGU133 Plus2.0 whole transcriptome data of 153 colonic biopsy samples(49 healthy adult, 49 adenoma, 49 CRC, 6 healthy children)(GEO accession numbers: GSE37364, GSE10714, GSE4183, GSE37267). Whole promoter methylation analysis of genes showing inverse DNA methylationgene expression data was performed on 30 colonic samples using methyl capture sequencing.RESULTS Fifty-seven age-related Cp G sites including hypermethylated PPP1R16 B, SFRP1, SYNE1 and hypomethylated MGP, PIPOX were differentially methylated between CRC and normal tissues(P < 0.05, ?β≥ 10%). In the adenoma vs normal comparison, 70 CpG sites differed significantly, including hypermethylated DKK3, SDC2, SFRP1, SYNE1 and hypomethylated CEMIP, SPATA18(P < 0.05, ?β≥ 10%). In MS-HRM analysis, the SFRP1 promoter region was significantly hypermethylated in CRC(55.0% ± 8.4 %) and adenoma tissue samples(49.9% ± 18.1%) compared to normal adult(5.2% ± 2.7%) and young(2.2% ± 0.7%) colonic tissue(P < 0.0001). DNA methylation of SFRP1 promoter was slightly, but significantly increased in healthy adults compared to normal young samples(P < 0.02). This correlated with significantly increased SFRP1 m RNA levels in children compared to normal adult samples(P < 0.05). In CRC tissue the mR NA expression of 117 agerelated genes were changed, while in adenoma samples 102 genes showed differential expression compared with normal colonic tissue(P < 0.05, logF C > 0.5). The change of expression for several genes including SYNE1, CLEC3 B, LTBP3 and SFRP1, followed the same pattern in aging and carcinogenesis, though not for all genes(e.g., MGP). CONCLUSION Several age-related DNA methylation alterations can be observed during CRC development and progression affecting the m RNA expression of certain CRC- and adenoma-related key control genes.

Clinicopathological significance of LRP16 protein in 336 gastric carcinoma patients

AIM： To investigate the expression of leukemia related protein 16 （LRP16）, and the possible relationship between LRP16 expression and clinicopathological indices in 336 gastric carcinoma patients. METHODS： Immunohistochemistry was used to detect LRP16 expression in 336 cases of paraffin-embedded gastric carcinoma tissues and 60 cases of distal normal mucosa. The relationships between LRP16 expression and patients＇ age, tumor size, histological grade, clinical stage, metastatic status and prognosis were analysed. RESULTS： The expression of LRP16 was 58.6% （197/336） in gastric carcinoma and 31.7% （19/60） in distal normal gastric mucosa. The expression of LRP16 in carcinoma was significantly higher than that in normal mucosa tissues （x^2 = 14.929, P = 0.001）. LRP16 protein expression was found in 44.1% （63/143） carcinomas at stage Ⅰ and Ⅱ, and 69.4% （134/193） carcinomas at stage Ⅲ and Ⅳ （Z2 = 21.804, P = 0.001）, and in 56.9% （182/320） of cancers without metastasis but 93.8% （15/16） of those with metastasis （2 = 8.543, P = 0.003）. The expression of LRP16 was correlated with tumor size, infiltrative depth, clinical stage, lymphatic invasion and distant metastasis （all P 〈 0.05）. Follow-up data showed that there was a significant difference in median survival time between cancer patients with expression of LRP16 （27.0 mo） and those without （48.0 mo, Log rank =31.644, P = 0.001）. CONCLUSION： The expression of LRP16 may be associated with invasion, metastasis and prognosis of gastric cancer.

Expression of telomerase gene and apoptosis related genes in benign and malignant breast lesion

Objective:To analyse the expression of telomerase and apoptosis related protein,and explore the possible mechanism of breast cancer development.Methods:Immunohistochemistry method(SP)was used to detect the expression of hTERT,p53 and bcl-2 in the tissues of 48 cases of human breast cancer and 42 cases of benign lesions in breast.Results: The positive rates of expression of hTERT,p53 and bcl-2 in breast cancer were 87.50%,56.25%and 54.17%,respectively. Compared with the groups of adjacent noncancerous and benign lesions,there was a significant difference among three types of tissues(P<0.05).The positive rates of expression of p53 and bcl-2 in the group with positive expression of hTERT were 64.28%and 61.90%,respectively,and their difference was significant compared with the negative group(P<0.05). Conclusion:There is a correlation between the activation of telomerases and p53 gene mutation in the development of breast cancer,and they are perhaps relation to the down regulation of bcl-2.

Yinchenhao decoction attenuates obstructive jaundice-induced liver injury and hepatocyte apoptosis by suppressing protein kinase RNA-like endoplasmic reticulum kinase-induced pathway

BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(ER)stress is one of the signaling pathways that induce apoptosis.Moreover,the protein kinase RNA-like endoplasmic reticulum kinase(PERK)-induced apoptotic pathway is the main way;but its role in liver injury remains unclear.Yinchenhao decoction(YCHD)is a traditional Chinese medicine formula that alleviates liver injury and apoptosis,yet its mechanism is unknown.We undertook this study to investigate the effects of YCHD on the expression of ER stress proteins and hepatocyte apoptosis in rats with obstructive jaundice(OJ).AIM To investigate whether YCHD can attenuate OJ-induced liver injury and hepatocyte apoptosis by inhibiting the PERK-CCAAT/enhancer-binding protein homologous protein(CHOP)-growth arrest and DNA damage-inducible protein 34(GADD34)pathway and B cell lymphoma/leukemia-2 related X protein(Bax)/B cell lymphoma/leukemia-2(Bcl-2)ratio.METHODS For in vivo experiments,30 rats were divided into three groups:control group,OJ model group,and YCHD-treated group.Blood was collected to detect the indicators of liver function,and liver tissues were used for histological analysis.For in vitro experiments,30 rats were divided into three groups:G1,G2,and G3.The rats in group G1 had their bile duct exposed without ligation,the rats in group G2 underwent total bile duct ligation,and the rats in group G3 were given a gavage of YCHD.According to the serum pharmacology,serum was extracted and centrifuged from the rat blood to cultivate the BRL-3A cells.Terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling(TUNEL)assay was used to detect BRL-3A hepatocyte apoptosis.Alanine aminotransferase(ALT)and aspartate transaminase(AST)levels in the medium were detected.Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were used to detect protein and gene expression levels of PERK,CHOP,GADD34,Bax,and Bcl-2 in the liver tissues and BRL-3A cells.RESULTS Biochemical assays and haematoxylin and eosin staining suggested severe liver function injury and liver tissue structure damage in the OJ model group.The TUNEL assay showed that massive BRL-3A rat hepatocyte apoptosis was induced by OJ.Elevated ALT and AST levels in the medium also demonstrated that hepatocytes could be destroyed by OJ.Western blot or qRT-PCR analyses showed that the protein and mRNA expression levels of PERK,CHOP,and GADD34 were significantly increased both in the rat liver tissue and BRL-3A rat hepatocytes by OJ.The Bax and Bcl-2 levels were increased,and the Bax/Bcl-2 ratio was also increased.When YCHD was used,the PERK,CHOP,GADD34,and Bax levels quickly decreased,while the Bcl-2 levels increased,and the Bax/Bcl-2 ratio decreased.CONCLUSION OJ-induced liver injury and hepatocyte apoptosis are associated with the activation of the PERK-CHOP-GADD34 pathway and increased Bax/Bcl-2 ratio.YCHD can attenuate these changes.

Phosphatase and tensin homology deleted in chromosome 10,hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression

To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein expression in the patients with human colorectal adenomas and adenocarcinomas.Methods The expression of PTEN,HIF-1 alpha gene was detected by using in situ hybridization,and the VEGF expression levels by immunohistochemistry in colorectal adenomas and primary colorectal adenocarcinoma.Results Strong expression of HIF-1 alpha was detectable in the majority of colorectal dadenocarcinoma,particularly surrounding areas of necrosis in adenocarcinoma.PTEN,HIF-1 alpha mRNA and VEGF protein were positive in 51.6%,67.7% and 59.7% respectively in 62 cases of adenocarcinomas,and 77.8%,44.4% and 33.3% respectively in 18 cases of adenomas.The positive rate of VEGF was higher in the patients with colorectal adenocarcinomas than that in those with adenomas,whereas that of PTEN mRNA was contrary.HIF-1 mRNA expression was correlated significantly with lymph node metastasis,liver metastasis,Duke’s stage and recurrence.During colorectal tumor progression,the expression of HIF-1 alpha mRNA was positively correlated with the VEGF protein expression (χ2= 4.751 ,P<0.05),but negatively with the PTEN mRNA expression(χ2=21.84,P<0.01).Conclusion The absence or low expression of PTEN and the increased levels of HIF-1α and VEGF may paly an important role in carcinogenesis and progression of colorectal carcinoma.These results suggest that VEGF upregulated by HIF-1 alpha gene may be involved in angiogenesis of colorectal adenocarcinoma.4 refs,1 tab.

Clinicopathological significance and prognostic value of LRP16 expression in colorectal carcinoma

AIM: To explore the expression of leukemia related protein 16 (LRP16) in colorectal carcinoma, and analyze its correlation with clinicopathologic features and prognosis. METHODS: Immunohistochemistry for LRP16 was performed in 201 cases of colorectal carcinoma and 60 cases of distal normal mucosa. Medical records were reviewed and clinicopathological analysis was performed. RESULTS: LRP16 expression was detected in 117 of 201 cases of the colorectal carcinoma and in 21 cases of 60 distal normal mucosa. The expression of LRP16 in carcinoma was significantly higher than that in normal mucosa (χ 2 = 9.999, P = 0.002). LRP16 protein expression was found in 43.3% (52/120) of carcinoma at stage Ⅰ and Ⅱ , and 80.2% (65/81) of carcinoma at stage Ⅲ and Ⅳ (χ 2 =27.088, P = 0.001). Correlation between LRP16 expression and clinicopathological factors was significant in differentiation (P = 0.010), tumor size (P = 0.001), infiltrative depth (P = 0.000) and distant metastasis (P = 0.027). The difference of median survival time between cancer patients with LRP16 expression (38.0 mo) and those without was statistically significant (105.0 mo, Log rank = 41.455, P = 0.001). The multivariate survival analysis revealed that LRP16 expression was correlated significantly (Cox’s regression: P = 0.001, relative risk = 2.082) with shortened survival in the patients with colorectal cancer. CONCLUSION: The expression of LRP16 is related to the degree of differentiation, invasiveness, metastasis and prognosis of colorectal carcinoma.

Emerging therapies in gastrointestinal cancers

Members of the receptor tyrosine kinase family, that include EGFR, ErbB-2/HER-2, ErbB-3/HER-3 and ErbB-4/ HER-4, are frequently implicated in experimental models of epithelial cell neoplasia as well as in human cancers. Therefore, interference with the activation of these growth factor receptors represents a promising strategy for de- velopment of novel and selective anticancer therapies. Indeed, a number of inhibitors that target either EGFR or HER-2, with the exception of a few that target both; have been developed for treatment of epithelial cancers. Since most solid tumors express different ErbB receptors and/or their ligands, identification of inhibitor(s), targeting multiple EGFR family members may provide a therapeutic benefit to a broader patient population. Here we describe the significance of an ErbB family of receptors in epithelial cancers, and summarize different available therapeutics targeting these receptors. It also emphasizes the need to develop pan-ErbB inhibitors and discusses EGF-Receptor Related Protein, a recently isolated negative regulator of EGFR as a potential pan-ErbB therapeutic for a wide vari- ety of epithelial cancers.

印度紫檀营养贮藏蛋白质的主要特性

采用光学和电子显微镜技术、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术分离鉴定热带落叶树木印度紫檀营养贮藏蛋白质(VSPs),并采用免疫印迹技术分析包括印度紫檀在内的5种热带树木VSPs之间的免疫相关性。结果表明:印度紫檀VSPs包括4种蛋白质,分子质量分别为33.5、28、21和19.5ku;它们主要分布在枝条木薄壁细胞、木射线薄壁细胞、韧皮薄壁细胞和韧皮射线薄壁细胞、皮层薄壁细胞以及树干、大根的韧皮薄壁细胞和韧皮射线薄壁细胞中;它们的超微结构存在明显差异,大致可以分为电子致密的团块状、絮状和均一状3种形态,不同形态的VSPs存在于不同的贮藏蛋白质细胞中,具有细胞特异性;在新梢发育时期,积累在枝条中的VSPs明显消失,但积累在树干和大根中的VSPs和落叶期相比没有明显变化,积累在枝条中的营养贮藏蛋白质是新梢生长发育所需氮素的主要来源;同一科树木的营养贮藏蛋白质之间存在较强的免疫相关性,但不同科树木的营养贮藏蛋白质之间缺乏免疫相关性。树木营养贮藏蛋白质生化性质的多样性可能反映了生物学功能的多样性。

Characterization and Proteomic Analysis of Novel Rice Lesion Mimic Mutant with Enhanced Disease Resistance

Lesion mmic mutants(LMMs)are plants that spontaneously form lesions without pathogeninfection or external stimulus and exhibit resistance to pathogens.Here,a rice LMM was created by ethylmethane sulfonate mutagenesis,named as hpil(hydrogen peroxide induced lesion).Diaminobenzidineand trypan blue staining showed that large amounts of H_(2)O_(2) were produced and cell death was occurredat and around the parts of lesion mimic in the rice leaves.The phenotype of hpil is controlled by a singlerecessive gene,localized at a 2 Mb interval on chromosome 2.The data suggested that hpil is a novelLMM with enhanced bacterial and fungal disease resistance,and multiple pathogenesis-related proteins(PRs)were up-regulated.The proteomes of leaves at three positions(different degrees of lesion mimicseverity)were characterized in hpil compared with its wild type plant.Differentially expressed proteinswere detected by two dimensional difference gel electrophoresis and 274 proteins were identified byMALDITOF/TOFTM.These proteins were related to metabolic process,cellular process and response tostimulus,with mostly down-regulated in hpil leaves.Many of these proteins were related to the Calvincycle,photosynthetic electron transport chain,glycolysis/gluconeogenesis and phosphonates pathways.Some resistance-related proteins including 14-3-3 proteins,OsPR10 and antioxidases such asperoxidase,superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesionmimic.These results provide the foundation for cloning of the target gene and shed light on themechanism involved in autaimmunity of rice.

Efficacy of MRP8/14 as a Marker of Disease Activity in Rheumatoid Arthritis

Objective: Early and accurate evaluation of the presence and activity of synovitis is extremely important in the diagnosis and treatment of rheumatoid arthritis. Myeloid related protein 8/14 (MRP8/14), also known as calprotectin or S100A8/A9 is considered as a sensitive marker for local inflammatory activity in rheumatoid arthritis. The aim of this study is to demonstrate the efficacy of MRP8/14 as a marker of disease activity in RA. Methods: Thirty-one patients with diagnosis of RA who received treatment without biological drugs at our institution were included in this study. Serum MRP8/14, CRP and MMP-3 were tested in all patients. Disease activity was evaluated using DAS28-CRP and SDAI. Ultrasonography was performed on the wrists and MCP joints of both hands using semi-quantitative scale of power Doppler signal. The sum of scales in joints was calculated as the PD score. The correlation of MRP8/14 with serum biomarkers, disease activity and ultrasonography examination was investigated. Result: Serum MRP8/14 was strongly correlated with CRP (r = 0.63) and MMP-3 (r = 0.69). A correlation was observed between serum MRP8/14 and DAS28-CRP (r = 0.53) and SDAI (r = 0.66). No significant correlation was found between PD scores and MRP8/14. Conclusion: This study demonstrated that MRP8/14 is correlated with evaluated disease activity and markers of serum inflammatory response in patients not using biological drugs. MRP8/14 is considered an effective new method for objective evaluation of synovitis in RA.

Cell cycle arrest effect of compound YSY-01A, a new proteasome inhibitor,on SK-OV-3 cells

Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aimed to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human ovarian cancer SK-OV-3 cells. The results suggested that YSY-01A significantly （P〈0.05） inhibited cellular proliferation of SK-OV-3 cells in a concentration-dependent and time-dependent manner. Furthermore, YSY-01A induced a G2/M cell cycle arrest of SK-OV-3 cells. Further investigation revealed that YSY-01A significantly （P〈0.05） changed the expression levels of a series of cell cycle related protein, such as cyclin B1, cdc2, and p-cdc2 （T14）. Meanwhile, YSY-01A could inhibit the TNF-a-induced NF-kB nuclear translocation and lead to the increase of 1kBa as well as the decrease of IKK and Gadd45a In conclusion, YSY-01A showed remarkable anti-cancer activity on SK-OV-3 cells, and its molecular mechanisms were related to G2/M cell cycle arrest.

Cell cycle arrest effect of compound YSY-01A, a new proteasome inhibitor, on HT-29 cells in vitro

Compound YSY-01A, a recently synthesized proteasome inhibitor, has shown potent growth-inhibitory effect on tumor cells in previous researches. However, the mechanism of its inhibitory effects, especially on cell cycle, remains largely unclear. This study aims to evaluate the correlation between cell cycle arrest effect of YSY-01A and its anti-cancer effect, and to probe the possible molecular mechanisms for its effects on human colorectal adenocarcinoma cells HT-29. The results suggested that YSY-01A significantly （P0.05） inhibited cellular proliferation of HT-29 cells in a time and concentration-dependent manner. Furthermore, YSY-01A suppressed the G 2 /M transition of HT-29 cells, whereas the mitotic inhibitor paclitaxel induced M phase accumulation. Further investigation revealed that YSY-01A significantly （P0.05） up-regulated the expression levels of a series of cell cycle related protein, such as cyclin B1, Wee1, p-cdc2 （Tyr15）, p53, p21, and p27. The HT-29 cells only exhibited typical cytotoxic symptom when YSY-01A concentration reached 0.5 μM （P0.05）, which was above the dose we used in the mechanism research. In conclusion, YSY-01A showed remarkable anti-cancer activity on HT-29 cells, and its molecular mechanisms are related to G 2 /M cell cycle transition arrest.

Association of common polymorphisms in the LRP6 gene with sporadic coronary artery disease in a Chinese population

Background Genetic factors contribute to the development of coronary artery disease （CAD）.Recently,a missense mutation in the low density lipoprotein receptor related protein 6 （LRP6） gene,encoding low density lipoprotein receptor related protein 6,has been implicated in an autosomal dominant form of early-onset CAD.The aim of this study was to determine whether the common variants in LRP6 are associated with sporadic CAD in Chinese.Methods A total of 766 CAD patients and 806 healthy controls were included in this study.The presence of angiographic CAD was determined by coronary angiographic analysis.Six signal nucleotide polymorphisms （SNPs） were genotyped using the polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） technique.Results A significant association was detected between rs11054731 in LRP6 intron 2 and CAD in our cohort （P=0.001）.The CC genotype and C allele frequency in the case group were 52% and 72%.Using a dominant model of inheritance,the C allele of rs11054731 was shown to be an independent risk factor for CAD with an OR of 1.45 （95% CI：1.19-1.77,P=0.0002）.With the stratification according to the number of affected coronary arteries,an association was observed between rs11054731 and CAD （P=0.0002）.No significant association was observed between any other SNPs and the risk of CAD.Conclusion The C allele of the rs11054731 within the LRP6 gene was associated with increased risk and extent of CAD in Chinese.