Acute liver failure and end-stage liver diseases are llfe-threatening.So far,orthotopic liver transplantation is the unique treatment available for these late-stage liver diseases,which,however,is limited by the lack of donor liver.While hepatocyte transplantation,bioartificial liver and tissue engineered liver are potential alternative treatments,the limited availability of functional hepatoeytes is the major hurdle.Due to their extensive capacity for self-renewal and pluripotency to differentiate into cells,embryonic stem cells represent a potential unlimited cell source for therapy.The clinical application of embryonic stem cells in liver diseases requires well-defined and efficient protocols for differentiation,characterization and purification in vitro.Hepatic differentiation of embryonic stem cells in different culttire systelns and the problems remained are reviewed in this article.

Objective; To study the effect of embryos on the regulation of integrin ?3 in cultured mice endometrial cells with a three-dimensional model. Methods: The mice blastocysts on day 4 of pregnany were cultured on a three-dimensional endometrium in vitro,the level of integrin ?3 mRNA was measured with RT-PCR and prtein measured with immuno-blot method. Results:The level of integrin ?3 mRNA and protein expression was significantly increased by embryos( 1.021?0.022 vs 0.434?0. 154,126. 83?8.27 vs 69.90?6.16) (P

Objective To screen potential serum HCC associated proteins with low molecular weight and low abundance for better understanding the pathological mechanism of HCC and discovering new biomarkers.Methods All serum samples were collected from 81 HBV-related HCC patients,43 chronic hepatitis B patients and 36 cirrhosis patients.Serum protein fingerprint profiles were first generated by selected WCX2 protein chip integrating with SELDI-TOF-MS,and then normalized and aligned by Ciphergen SELDI Software 3.1.1 with Biomarker Wizard.Comparative analysis of the intensity of corresponding protein fingerprint peaks in normalized protein spectra was performed.Some protein peaks with significant difference among HCC,cirrhosis and chronic hepatitis B groups were found.The reproducibility of the SELDI system was assessed before serum protein fingerprint profiles analysis.Results The intra-and inter-assay CV for intensity and m/z in this SELDI system were 17.46% and 0.024%,and 17.74% and 0.024% respectively.Total 128 protein fingerprint peaks between 2 000 to 30 000 Da were identified under the condition of signal to noise＞5 and minimum threshold for cluster＞20%.Eighty-seven proteins were found to significantly expressed between HCC and cirrhosis groups(P＜0.05).Of the above differential proteins,forty-five proteins had changes greater than two fold,including 15 up-regulated proteins and 30 downregulated proteins in HCC sera.Between HCC and chronic hepatitis B groups,nine of fifty-two differential proteins(P＜0.05) had intensities of more than two folds,including 2 up-regulated proteins and 7 downregulated proteins in HCC sera.Between cirrhosis and chronic hepatitis B groups,twenty-eight of seventynine significantly differential proteins(P＜0.05) changed greater than two folds in intensity,including 17 up-regulated proteins in cirrhosis seru and 11 down-regulated proteins in chronic hepatitis B sera.Analysis of above leading differential proteins among three diseases using subtraction difference mode,the 5 common down-regulated proteins 2 870,3 941,2 688,3 165 and 5 483 m/z in HCC sera and 2 common up-regulated proteins 3 588 and 2 017 m/z in cirrhosis and HCC sera were screened.But no statistic difference in the level of protein 2 017m/z was found between HCC group and normal group inour previous study.Conclusion Because the interference of unspecific proteins from hepatitis B and cirrhosis could be eliminated partly in HCC sera through subtraction difference analysis,these 6 common differential proteins (2 870,3 941,2 688,3 165,5 483,3 588 m/z)have obvious advantages of increased specificity for evaluating the pathological state of HCC and might become promising candidate biomarkers in the diagnosis of HCC.

OBJECTIVETo compare gene expression profile of human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to screen for metastasis-related genes.

METHODSGene expression profile of MHCC97-L and HCCLM3, two HCC cell lines with similar genetic background but different in spontaneous metastatic potentials, were studied by cDNA microarray.

RESULTSFrom 1,626 screened genes, 25 differentially expressed genes were found, 18 showed decreased expression including the decreased expression of cell cycle control genes Rb2, mismatch repair gene hMSH2, and signal transduction gene protein kinase C beta 2 and 7 increased expression including signal transduction gene MAP kinase kinase 6, cell proliferation gene E25, immunity related gene SP40, 40, etc in HCCLM3.

CONCLUSIONThe genes, being closely associated with cancer metastasis, could be considered as potential markers to predict metastasis and targets for anti-metastasis intervention.

Blotting, Northern ; Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression ; Gene Expression Profiling ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Metastasis ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Neoplasm ; analysis ; Tumor Cells, Cultured

To evaluate the reagent 2-methoxy-4,5-dihydro-1H-imidazole used for isotopic labeling in quantitative proteomics, we synthesized 2-methoxy-4,5-dihydro-1H-imidazole and its tetradeuterated analog in three steps. Prior to tryptic cleavage, bovine serum albumin (BSA) was reduced and alkylated. Tryptic peptides were derivatized with an equal volume of either DO or D4 and D4-derivatized peptides were mixed with at variable ratio (from 10:1 to 1:5) prior to MS and MS/MS analysis. We used matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and Electro spray ionization-mass spectrometry (ESI-MS) to evaluate the quantitative capability of labeling. The specificity of the reagent is excellent: only lysine side chains were modified among tryptic peptides. MALDI and ESI ionization modes not only could achieve the quantification of differentially expressed proteins but also facilitate the de novo sequencing. This side-chain modification can be used for quantitative analysis with proteomic strategies involving liquid chromatography. Reverse phase liquid chromatography (RPLC) kept a good resolution, and the introduction of D atoms did not introduce a variation of retention time between heavy and light peptides in RPLC.
Imidazoles ; chemistry ; Isotope Labeling ; methods ; Lysine ; chemistry ; Peptides ; analysis ; chemistry ; Proteomics ; methods ; Serum Albumin, Bovine ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo identify specific serum glycoprotein profiles that correspond to the carcinogenic process of primary liver cancer (PLC) by analyzing a population with high-incidence of PLC using lectin affinity microarray.

METHODSSerum samples were collected from individuals classified as high risk for PLC (including patients with liver cirrhosis and hepatitis B) and development of PLC was recorded. Healthy individuals served as normal controls. The serum samples were subjected to glycoprotein profling by using lectin microarrays and the results were confirmed by lectin blot. Between-group differences were statistically analyzed.

RESULTSPLC carcinogenesis was found to be correlated with enhanced affinity for AAL, ACL, ConA, LCA, MPL, NML, PHA-E, PHA-L, PSA, RCA-I, STL, VAL,WGA, and SNA (P less than 0.05). These data implied that changes in specific glycan structures, such as aFuc, GlcNAc, GalNAc, mannose, bisecting GlcNAc and terminal beta1-4 Gal, may be involved in PLC carcinogenesis . The PLC group showed significantly different results for all detected lectins, except SNA (P less than 0.05). However, among the PLC group, the SNA affinity was not significantly different for the hepatitis B group (P =0.443, P more than 0.05).

CONCLUSIONGlycans may be associated with the carcinogenic process of PLC and may be developed as diagnostic and prognostic biomarkers of PLC in the future.

Carcinogenesis ; Chromatography, Affinity ; Cohort Studies ; Glycoproteins ; blood ; Humans ; Lectins ; blood ; Liver Neoplasms ; blood ; pathology

Objective:To analyze the efficacy of posterior staged correction in the treatment of severe kyphoscoliosis.Methods:Retrospective analysis was conducted on 61 patients with severe kyphoscoliosis who underwent one-stage posterior Ponte osteotomy followed by Halo-femoral traction and two-stage posterior correction in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School between January 2010 and January 2020. There were 23 males and 38 females with an average age of 22.0(18.0, 25.5) years. The etiologies were idiopathic in 26 cases, congenital in 17 cases, neuromuscular in 16 cases and Marfan syndrome with kyphoscoliosis in 2 cases. The curves were thoracic in 49 cases, thoracolumbar in 3 cases and double major in 9 cases. The apical vertebrae were T 5 level in 1 case, T 7 level in 2 cases, T 8 level in 9 cases, T 9 level in 15 cases, T 10 level in 23 cases, T 11 level in 8 cases, T 12 level in 1 case, and L 1 level in 2 cases. The flexibility of main curve was 13.5%±8.6%. The Cobb angle of main curve, global kyphosis (GK), coronal trunk shift (CTS), sagittal vertical axis (SVA), thoracic kyphosis (TK), lumbar lordosis (LL), pelvic incidence (PI), pelvic tilt (PT), and sacral slope (SS) were assessed at pre-operation, post-traction, post-operation and the last follow-up. The quality of life was evaluated using the MOS item short form health survey (SF)-36 questionnaire, and the complications during peri-operation and long-term follow-up were recorded. Results:All 61 patients were followed up for 25.0 (24.0, 26.5) months. The Cobb angle of main curve and GK were 121.4°±13.9° and 86.8°±20.0° at pre-operation, 94.1°±18.7° and 66.9°±15.3° at post-traction, 78.5°±20.3° and 54.7°±13.6° at post-operation and 79.5°±20.1° and 53.2°±11.3° at the last follow-up, respectively. The differences were statistically significant ( F=210.54, P<0.001; F=93.74, P<0.001). There was no significant difference between the last follow-up and post-operation ( P>0.05). There was no significant correction loss of SVA, TK, LL, PI, PT or SS at the last follow-up when compared with those at post-operation ( P>0.05). The CTS was 17.1±9.8 mm at pre-operation, 17.5±11.4 mm at post-operation, 11.1 (5.9, 23.3) mm at the last follow-up and the difference was statistically significant (χ 2=6.70, P=0.035). The difference between the last follow-up and post-operation was statistically significant ( P=0.032). The scores of physical functioning 80.0 (75.0, 85.0), general health 82.0 (69.5, 87.0) and social functioning 75.0 (62.5, 75.0) in SF-36 at the last follow-up were significantly improved compared with those at pre-operation ( Z=-2.11, P=0.035; Z=-3.64, P<0.001; Z=-2.07, P=0.039). During the traction process, the complications included pin track infection in 1 case, deep vein thrombosis of the lower extremity in 2, misplacement of pedicle screws in 3, coronal decompensation at immediate post-operation in 2, sagittal decompensation at immediate post-operation in 1, and 1 patient had broken rod at 3 years follow-up, respectively. Conclusion:The posterior staged correction could provide satisfying radiographic and clinical outcomes in patients with severe kyphoscoliosis, which could be well maintained during 2 years follow-up. Therefore, the posterior staged correction is a safe and effective treatment for severe kyphoscoliosis.