Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of prostate cancer

Aim： To identify the serum biomarkers of prostate cancer （PCa） by protein chip and bioinformatics. Methods： Serum samples from 83 PCa patients and 95 healthy men were taken from a mass screening in Changchun, China. Protein profiling was carried out using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF MS）. The data of spectra were analyzed using two bioinformatics tools. Results： Eighteen serum differential proteins were identified in the PCa group compared with the control group （P 〈 0.01）. There were four proteins at the higher serum level and 14 proteins at the lower serum level in the PCa group. A decision tree classification algorithm that used an eight-protein mass pattern was developed to correctly classify the samples. A sensitivity of 92.0 % and a specificity of 96.7 % for the study group were obtained by comparing the PCa and control groups. Conclusion： We identified new serum biomarkers of PCa. SELDI-TOF MS coupled with a decision tree classification algorithm will provide a highly accurate and innovative approach for the early diagnosis of PCa. （Asian J Androl 2006 Jan; 8： 45-51）

Prostate cancer： an emerging threat to the health of aging men in Asia

The aim of this study was to determine and examine the possible reasons for the difference in prostate cancer incidence between Asian men and North American men by literature review. Data regarding cancer incidence and mortality were obtained from the database of the International Agency for Research on Cancer （IARC）. A literature review was conducted by studying related articles published in peer-reviewed journals such as the The New England Journal of Medicine, Journal of Clinical Oncology, A Cancer Journal for Clinicians and Asian Journal of Andrology. To evaluate the early diagnosis and survival rates, the mortality.to-incidence rate ratio （MR/IR） was calculated from the IARC data. By comparing prostate cancer data between Asian men and North American men, we found that differences in the incidence rate and MR/IR could be attributed largely to a lack of annual prostate cancer screening with serum prostate-specific antigen （PSA） in most Asian countries. It is likely that PSA screening also contributes significantly to the differences in prostate cancer mortality rates. Prostate cancer has the highest incidence rate among five common malignancies in Asian Americans. However, the MR/IR ratio of prostate cancer is the lowest among cancers. These data seem to further support the usefulness of PSA screening, even though the percentage of low risk cancers is greater in prostate cancer than in other cancers. The low incidence rate of prostate cancer does not reflect the actual statistics of this disease in Asia. The data from limited institutions in many Asian countries seem to bias the true incidence and mortality rates. To improve this situation, incorporating PSA screening for prostate cancer, as well as constructing a nationwide cancer registration system, will be helpful.

Cloning and characterization of porcine aquaporin 1 water channel expressed extensively in gastrointestinal system

瞄准：在胃肠的系统克隆并且描绘猪的 aquaporins (AQP ) 。方法：基于 PCR 的克隆策略和赛跑被用来克隆从相对地抄录的猪肝 cDNA 编码顺序的全身的 AQP。散布的站流动光和一个基于 YFP 的荧光方法被用来测量红血球的渗透的透水性并且稳定地 transfected CHO 房间。RT-PCR，北污点，和免疫组织化学被用来决定克隆的 AQP 的胃肠的表示和本地化。在 transfected 房间和红细胞的蛋白质表示被西方的污点分析。结果：编码 271 氨基酸的 813 bp cDNA 猪的 aquaporin (指明的 pAQP1 ) 从肝 mRNA 被克隆(pAQP1 与人的 AQP1 有 93% 身份并且包含在 AQP 家庭，保存的二个 NPA 主题为连接 N 的 glycosylation 的一个一致序列，和在半胱氨酸 191 点的一个水银敏感的地点) 。RT-PCR 分析在猪的消化的腺和内脏揭示了 pAQP1 mRNA 的广泛的表示。北污点在选择消化机关给一个单身者看了 3.0 kb 抄本。pAQP1 蛋白质在小肠，唾液的腺的 microvessles，以及由 immunoperoxydase 的肝内胆汁管的上皮的中央 lacteals 是局部性的。是的高渗透的透水性禁止由 HgCl2 能稳定地在猪的红血球和 CHO 房间被检测有 pAQP1 cDNA 的 transfected。猪的红血球和 pAQP-transfected CHO 房间的 Immunoblot 分析揭示了 unglycosylated 28 ku 乐队和更大的 glycosylated 蛋白质。结论：pAQP1 是到目前为止能分子地被识别的第一猪的 aquaporin。在猪的消化机关的上皮和内皮细胞层的 pAQP1 的宽广分发可以在液体分泌物 / 吸收以及在胃肠的系统的消化功能和病理生理学建议调停隧道的水运输的一个重要角色。

羟基磷灰石纳米粒子携带Si-Star3质粒抑制小鼠前列腺癌生长的体外研究

DNA质粒介导的Stat3特异性RNA干扰呵以阻断Stat3信号并抑制前列腺肿瘤的生长。然而，抗肿瘤效心依赖于si—Stat3质粒的传递效率。本研究揭示了羟基磷灰石携带Stat3siRNA质粒对小鼠前列腺癌生长的影响。采用RM-1瘤块种植C57BL／6小鼠。CaCl2修饰的径基磷灰石携带si-Stat3质粒瘤体注射，测量肿瘤体积并绘制生长曲线，检测肿瘤组织学变化。采用RT-PCR和Westernblot分析Stat3展因与蛋白的表达变化，及pTyr—Stat3，Bcl-2，Bax，Caspase3，VEGF和cyclinD1蛋白的表达变化。采用免疫组织化学和TUNEL检测细胞凋亡。移植RM1瘤块C57BL／6小鼠，羟基磷灰石纳米粒子携带si-Stat3组与对照组相比较，显著抑制了肿瘤的生长，抑制率达到74％（P〈0．01）。在si-Stat3组肿瘤组织中，Stat3的表达水平显著下调，同时，肿瘤细胞的凋亡显著增加，凋亡指数达42％（P〈0．01）。肿垴组织中Stat3下游基因Bcl-2，VEGF和cyclinD1的表达水平也出现明显下调，同时Bax蛋白的表达增加和Caspase3的活性增强。这些结果表明，羟基磷灰石纳米粒子可以携带RNA干扰质粒进入肿瘤细胞内。

Renal protective effect of Ganoderma lucidum

OBJECTIVE Non-alcoholic fatty liver disease(NAFLD) encompasses a series of patho.logic changes ranging from steatosis to steatohepatitis,which may progress to cirrhosis and hepatocel.lular carcinoma.The purpose of this study was to determine whether Ganoderma lucidum polysaccha.ride peptide(GLPP) has therapeutic effect on NAFLD.METHODS ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD.Key metabolic path.ways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blotting.Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD.RESULTS GLPP administrated for a month alleviated hepatosteatosis,dyslipidemia,liver dysfunction and liver insulin resistance.Pathways of glycerophospholipid metabolism,fatty acid metabolism and primary bile acid biosynthesis were involved in the therapeutic effect of GLPP on NAFLD.Detection of key enzymes revealed that GLPP reversed low expression of CYP7 A1,CYP8 B1,FXR,SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice.Besides,GLPP inhibited fatty acid synthesis by reducing the expression of SREBP1 c,FAS and ACC via a FXR-SHP dependent mechanism.Additionally,GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepato.cytes induced by oleic acid and palmitic acid.CONCLUSION GLPP significantly improves NAFLD via regulating bile acid synthesis dependent on FXR-SHP/FGF pathway,which finally inhibits fatty acid synthesis,indicating that GLPP might be developed as a therapeutic drug for NAFLD.

Ganoderic acid A from Ganoderma lucidumretard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway

OBJECTIVE Autosomal dominant polycystic kidney disease(ADPKD) is a common.monogenetic disease characterized by progressive development of renal cysts.Thereis still further need for effective therapy.Based on our precious study that Ganoderma triterpenes(GT),which is the major secondary metabolites of Ganoderma lucidum,is able to attenuate renal cyst development.The aim of this study was to investigate the effect of a monomer,Ganoderic acid A(GA-A) that was purified from the GT,which has been reported to exhibit antinociceptive,antioxidative,hepatoproctive and anti.cancer activities,to have a potent anti-cyst effect in ADPKD.METHODS We first evaluated the potential cytotoxicity of GA-A on MDCK cells using a CCK-8 assay.Then we used MDCK cyst model,cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells.MDCK cells were co-incu.bated with 10 μmol·L^(-1) FSK with or without GA-A(25 μg·mL-1) and equal concentration GT as positive control from day 0 to day 6 to investigate the inhibitory effect of GA-A on cyst formation.And to further investigate the inhibitory effect of GA-A on cyst enlargement,MDCK cysts were treated with different concentration of GA-A(6.25,25 and 100 μg·mL-1) from day 5 to day 12.Next we used an embryonic kidney cyst model,wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP to prove the renal cyst inhibition at organ level.Meanwhile,we explored the possible mechanisms underlying GA-A inhibition on renal cyst development using MDCK cells treated with 10 μmol·L^(-1) FSK co-incubated with GA-A(25 μg·mL-1) and equal concentration GT.Several key components of Ras/MAPK pathway was evaluated by Western blot,the protein expression of H-ras,B-raf,p-ERK,Egr-1 and c-fos was evaluated.RESULTS MDCK cell viability was not affected by GA-A that were used ofincreasing concentrations up to 200 μg·mL^(-1).GA-A had no significant influence on cyst formation,but inhibited cyst enlargement dose-dependently and the inhibitory effect is significantly better than that of the same concentration of GT which proved that GA-A may be an effective monomer from GT.And after washing out GA-A on day 8,MDCK cysts regrew to a large size,suggesting that the inhibitory effect of GA-A on MDCK cyst enlargement was reversible.GA-A inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GA-A cyst inhibition at organ level,which is also more effective than equal concentration GT.Treatment of MDCK cells with FSK caused a significant elevation of H-ras,B-raf,p-ERK,Egr-1 and c-fos signaling molecules,while treatment with GA-A reduced the level of H-ras,B-raf,p-ERK,Egr-1 and c-fos expression.GA-A downregulated Ras/MAPK signaling pathway could contribute to its inhibitory effect on cyst development.CONCLUSION Ganoderic acid A from Ganoderma lucidum retard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway.

Ganoderma triterpenes slow cyst growth in polycystic kidney disease

OBJECTIVE Autosomal dominant polycystic kidney disease(ADPKD)is a common inherited disease with a high morbidity around 1/1000-1/400,characterized by progressive enlargement of fluid-fil ed cysts derived from renal tubular epithelial cells.Massive cysts gradually compress renal parenchyma destroying normal renal structures and compromising renal functions.Unfortunately,it will cause end-stage renal disease in most of the patients but without effective therapy now,who have to live on hemodialysis or kidney transplantation.Based on this present situation,it is of great significance to find early intervention to inhibit renal cyst development.The projective of this study was to investigate whether Ganoderma triterpenes(GT)can inhibit renal cyst development and study the related mechanism.METHODS and RESULTS First,we used MDCK cyst model,cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells.GT inhibited MDCK cyst formation significantly,and inhibited cyst enlargement dose-dependently proving GT cyst inhibition in vitro.Then we used an embryonic kidney cyst model,wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-c AMP.GT inhibited embryonic kidney cyst development significantly in a dosedependent and reversible manner proving GT cyst inhibition at organ level.Furthermore,we used two ADPKD mouse models with severe cystic kidney disease phenotypes.GT dramatically inhibited renal cyst development,decreased ADPKD mouse kidney volume and the cyst index inside proving GT cyst inhibition in vivo.By Western blot,we proved GT down-regulated Ras/MAPK signal pathway without detectable effect on m TOR signal pathway both in MDCK cells and two ADPKD mouse kidneys.CONCLUSION GT retard renal cyst development both in vitro and in vivo significantly.The related mechanisms were involved in GT promoting renal tubular epithelial cell differentiation,down-regulating intracellular c AMP level and Ras/MAPK signal pathway.