Advances in cardiovascular-related biomarkers to predict diabetic peripheral neuropathy.

Diabetic peripheral neuropathy (DPN) is a common chronic complication of diabetes mellitus. One of the most common types is distal symmetric poly-neuropathy, which begins as bilateral symmetry pain and hyperesthesia and gradually progresses into hypoesthesia with nerve fibre disorder and is frequently accompanied by depression and anxiety. Notably, more than half of patients with DPN can be asymptomatic, which tends to delay early detection.

Testosterone regulates the autophagic clearance of androgen binding protein in rat Sertoli cells.

Dysregulation of androgen-binding protein (ABP) is associated with a number of endocrine and andrology diseases. However, the ABP metabolism in Sertoli cells is largely unknown. We report that autophagy degrades ABP in rat Sertoli cells, and the autophagic clearance of ABP is regulated by testosterone, which prolongs the ABP biological half-life by inhibiting autophagy.

Renal tissue thawed for 30 minutes is still suitable for gene expression analysis.

Some biosamples obtained from biobanks may go through thawing before processing. We aim to evaluate the effects of thawing at room temperature for different time periods on gene expression analysis. A time course study with four time points was conducted to investigate the expression profiling on 10 thawed normal mice renal tissue samples through Affymetrix GeneChip mouse gene 2.

NANOG promotes liver cancer cell invasion by inducing epithelial-mesenchymal transition through NODAL/SMAD3 signaling pathway.

NANOG is a major transcription factor essential to the stem cell self-renewal and is associated with tumor malignancy, but the NANOG signaling in cancer metastasis is still elusive. In this study, we determined the expression of NANOG in hepatocellular carcinoma (HCC) and investigated its underlying mechanism in the metastasis of HCC. The expression levels of NANOG were examined in tumor tissues by immunohistochemistry.

[Applicability of the multiplex quantitative antibody array system for early diagnosis of hepatocellular carcinoma].

Objective: To develop an early and accurate detection method for hepatocellular carcinoma (HCC) based on detection of tumor-associated serum markers using a multiplex quantitative antibody array.

Methods: The double-antibody sandwich principle was used to establish an antibody array composed of eight cancer-related serum markers, including alpha-fetoprotein (AFP), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), transforming growth factor-beta 1 (TGF-b1), and vascular endothelial growth factor (VEGF). Serum samples from 160 cases of clinically diagnosed HCC and from 58 cases of liver cirrhosis (LC; controls) were obtained to test the array.

[Identification of metastasis-related osteopontin expression and glycosylation in hepatocellular carcinoma].

Objective: To test expression level and glycosylation level of OPN in HCC cell lines with different metastatic potential and HCC tissues, and investigate the correlation between the glycosylation change and the liver cancer transporting as well as its significance.

Methods: The level of OPN expression in liver cancer tissue(6 cases of non-metastasis and 7 cases of metastasis)as well as HCC cell lines with different metastatic potential (L02, Hep3B, MHCC97L, MHCC97H, HCCLM3, HCCLM6)was identified by immunohistochemistry and Western Blot, and then OPN was purified from HCC tissues by immunoprecipitation, followed by glycosylation detection of OPN from non-metastatic and metastatic HCC tissues by multiple lectin blot. Data were analyzed by t-test and variance analysis.

iTRAQ-2DLC-ESI-MS/MS based identification of a new set of immunohistochemical biomarkers for classification of dysplastic nodules and small hepatocellular carcinoma.

The study aims to develop novel clinical immunohistochemical biomarkers for distinguishing small hepatocellular carcinoma (sHCC) from dysplastic nodules (DN). iTRAQ-2DLC-ESI-MS/MS technique was used to screen immunohistochemical biomarkers between precancerous lesions (liver cirrhosis and DN) and sHCC. A total of 1951 proteins were quantified, including 52 proteins upregulated in sHCC and 95 proteins downregulated in sHCC by at least 1.

[Effects of AKR1B10 gene silence on the growth and gene expression of HCC cell line MHCC97H].

Objective: To explore the biological function and possible underlying mechanism of aldo-keto reductase family 1 member B10 (AKR1B10) gene during hepatocarcinogenesis.

Methods: A pair of chemically synthesized small interfering RNA (siRNA) targeting on AKR1B10 was transfected into liver cancer cell line MHCC97H by LipofectamineTM 2000. After confirming the interfering effects of AKR1B10-siRNAs through Quant SYBR Green polymerase chain reaction (Real-time PCR), Western blot and enzymatic activity assay, the capabilities of proliferation and apoptosis of the transfected cells were observed by CCK-8 assay and flow cytometry analysis, and the expressions of a group of tumor-related gene such as c-myc, c-fos, N-ras were observed through Real-time PCR.

Array-based profiling of the differential methylation status of CpG islands in hepatocellular carcinoma cell lines.

Alterations in the DNA methylation status particularly in CpG islands are involved in the initiation and progression of many types of human cancer. A number of DNA methylation alterations have been reported in hepatocellular carcinoma (HCC). However, a systematic analysis is required to elucidate the relationship between differential DNA methylation status and the characteristics and progression of HCC.

[Comparison of the serum proteomes of pathological stages during hepatocarcinogenesis].

Objective: To compare the 2-DE profiles for serum proteins of different pathological stages during hepatocarcinogenesis.

Methods: Sera from hepatocellular carcinoma patients, cirrhosis patients, chronic hepatitis patients and healthy controls were collected. After sonication, albumin and immunoglobulin (IgG) depletion, and desalination, sera were subjected to 2-DE, the differential protein spots were identified by MALDI-TOF-MS.

[Differentially expressed proteins in the precancerous stage of rat hepatocarcinogenesis induced by diethylnitrosamine].

Objective: To screen the differentially expressed proteins especially at the precancerous stage of diethylnitrosamine (DEN) induced hepatocarcinogenesis by comparative proteome research.

Methods: Rats were divided into normal and DEN groups and sacrificed periodically. The liver samples were stained with gamma-glutamyl transpeptidase (GGT) and HE to distinguish the preneoplastic lesion (pre-HCC) from the normal and HCC tissues.

[Protein profiles of multinodular hepatocellular carcinoma with multicentric occurrence or with intrahepatic metastasis].

Objective: To analyze the protein expression profiles of multinodular hepatocellular carcinoma (HCC) with multicentric occurrence (MO) or with intrahepatic metastasis (IM).

Methods: 5 IM and 6 MO patients were divided into groups of IM1, IM2, MO1 and MO2 according to the size of node of HCC. Two dimensional gel electrophoresis (2-DE) and mass spectrum were used to analyze the protein expression profiles.

Application of SELDI-TOF-MS to identify serum biomarkers for renal cell carcinoma.

Purpose: Early diagnosis is critical for improving the outcome of patients with renal cell carcinoma (RCC). In this study, we applied a proteomic approach to identify serum biomarkers associated with different stages of renal tumor development.

Materials And Methods: The protein expression profiles in patient serum samples were analyzed using surface enhanced laser desorption/ionization time-of-flight mass spectroscopy (SELDI-TOF-MS).

[Analysis of the expression of Slit/Robo genes and the methylation status of their promoters in the hepatocellular carcinoma cell lines].

Objective: To analyze the expression of genes in the Slit/Robo signaling pathway, and the methylation status of their promoters in hepatocellular carcinoma (HCC) cell lines.

Methods: Genomic DNA and total RNA were isolated from 9 HCC cell lines of different metastatic ability (Hep3B, HepG2, PLC/PRF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, LM3, LM6) and a control cell line L-02. The expression profiles of Slit1, Slit2, Slit3, Robo1, and Robo3 were analyzed by reverse transcription polymerase chain reaction (RT-PCR).

[Screening for differential methylation status by CpG island microarray in the hepatocellular carcinoma cell lines].

Objective: To profile the methylation alterations of CpG islands in hetpatocellular carcinoma cell lines.

Methods: A global analysis of DNA methylation using the Human CpG-island 12K Array (HCGI12K) from Canada University Health Network was performed on nine human hepatocellular carcinoma (HCC) cell lines (Hep3B, HepG2, PLC/RPF/5/RPF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, HCCLM3, HCCLM6) and a control cell line Chang's liver. Metastatic potential related alterations were also screened in MHCC97 series cell lines (MHCC97-H, MHCC97-L, HCCLM3, HCCLM6), using MHCC97-L, a cell line with low metastatic potential, as control.

[Analysis of gene expression profile during malignant transformation of rat liver oval-like cells].

Objective: To investigate the changes of gene expression profile during malignant transformation of rat liver oval-like cells and to analyze the significances of these changes.

Methods: MNNG initiated WB-F344 cells were exposed to H2O2 once a week (repeated 21 times) to induce their malignant transformation. The characteristics of the transformed cells were confirmed by morphology, genetics and soft agar assay.

[Proteomic analysis in combination with CT diagnosis to distinguish renal cell carcinoma from renal benign masses].

Objective: To identify the proteomic differences between renal cell carcinoma (RCC) and renal benign masses and to evaluate the diagnostic value of parallel and serial test combining with CT and surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS).

Methods: Serum samples were collected from 96 patients with renal tumors, 62 RCC cases and 34 renal benign mass cases, all of which had been evaluated by CT before surgery. The sera were analyzed using IMAC-Cu2+ ProteinChip system by SELDI-TOF-MS.

[Proteomic analysis of hepatitis B virus-infected human liver tissues].

Objective: To search for the difference of protein molecules expression of HBV infection.

Methods: Specimens taken from human normal liver tissues (group A), HBV infected human liver tissues which were HBsAg positive, and HBsAg, anti-HBe, and anti-HBc positive in serum (group B) were analysed through the methods of 2-dimensional electrophoresis (2-DE) and matrix-assisted laser-desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS).

Results: Totally 1125 plus/minus 56 (n=3) spots were detected in the sample of group A, 1203 plus/minus 42 (n=3) in group B samples.

Screening serum hepatocellular carcinoma-associated proteins by SELDI-based protein spectrum analysis.

Aim: To find out potential serum hepatocellular carcinoma (HCC)-associated proteins with low molecular weight and low abundance by SELDI-based serum protein spectra analysis, that will have much application in the diagnosis or differentiated diagnosis of HCC, as well as giving a better understanding of the mechanism of hepato-carcinogenesis.

Methods: Total serum samples were collected with informed consent from 81 HCC patients with HBV(+)/cirrhosis(+), 36 cirrhosis patients and 43 chronic hepatitis B patients. Serum protein fingerprint profiles were first generated by selected WCX2 protein chip capture integrating with SELDI-TOF-MS, then normalized and aligned by Ciphergen SELDI Software 3.

Identification of N-glycan of alpha-fetoprotein by lectin affinity microarray.

Purpose: The occurrence of cancer accompanies with changes in glycosylation of related proteins. A simple, rapid and high-throughput manner analyzing the N-glycan moiety is needed in the cancer glycoproteome study.

Methods: Gel-slide was used as solid support of lectin microarray.

[Screening low molecular weight protein biomarkers relevant to portal vein tumor thrombi in serum of patients with hepatocellular carcinoma].

Objective: To screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients.

Methods: Serum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software.

[Study of serum proteome biomarkers with relation to the formation of portal vein tumor thrombi in hepatocellular carcinoma patients].

Objective: To screen serum proteome biomarkers and establish predictive model with relation to the formation of portal vein tumor thrombi (PVTT) in hepatocellular carcinoma (HCC) patients.

Methods: Serum samples were collected from 135 HCC patients, which were divided, into training set (including 33 HCC patients with PVTT and 62 HCC patients without PVTT) and blind testing set (including 18 HCC patients with PVTT and 22 HCC patients without PVTT). Special serum protein or peptide pattern was determined by SELDI-TOF-MS measurement after treating the sample onto WCX2 protein chip for each case.

[Glutamate-neuroexcitotoxicity protective study of monoclonal antibody against human NMDA receptor key subunit].

Objective: To study if monoclonal antibody against human NMDA receptor key subunit (NR1) may protect neurons from excitotoxicity.

Methods: We cultured primary hippocampal neurons from 10 newborn rats and made the glutamate excitotoxicity model, examined the ratio of surviving neuron (Trypan blue dye staining) and LDH assay to study the protective effects of mAbN1, There were 2 plates in parallel per group, the experiments were repeated 4 times.

Results: 0.