Plasma Metabolic Profile Determination in Young ST-segment Elevation Myocardial Infarction Patients with Ischemia and Reperfusion: Ultra-performance Liquid Chromatography and Mass Spectrometry for Pathway Analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study was to establish a disease differentiation model for ST-segment elevation myocardial infarction (STEMI) youth patients experiencing ischemia and reperfusion via ultra-performance liquid chromatography and mass spectrometry (UPLC/MS) platform, which searches for closely related characteristic metabolites and metabolic pathways to evaluate their predictive value in the prognosis after discharge.</p><p><b>METHODS</b>Forty-seven consecutive STEMI patients (23 patients under 45 years of age, referred to here as "youth," and 24 "elderly" patients) and 48 healthy control group members (24 youth, 24 elderly) were registered prospectively. The youth patients were required to provide a second blood draw during a follow-up visit one year after morbidity (n = 22, one lost). Characteristic metabolites and relative metabolic pathways were screened via UPLC/MS platform base on the Kyoto encyclopedia of genes and genomes (KEGG) and Human Metabolome Database. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of characteristic metabolites in the prognosis after discharge.</p><p><b>RESULTS</b>We successfully established an orthogonal partial least squares discriminated analysis model (R2X = 71.2%, R2Y = 79.6%, and Q2 = 55.9%) and screened out 24 ions; the sphingolipid metabolism pathway showed the most drastic change. The ROC curve analysis showed that ceramide [Cer(d18:0/16:0), Cer(t18:0/12:0)] and sphinganine in the sphingolipid pathway have high sensitivity and specificity on the prognosis related to major adverse cardiovascular events after youth patients were discharged. The area under curve (AUC) was 0.671, 0.750, and 0.711, respectively. A follow-up validation one year after morbidity showed corresponding AUC of 0.778, 0.833, and 0.806.</p><p><b>CONCLUSIONS</b>By analyzing the plasma metabolism of myocardial infarction patients, we successfully established a model that can distinguish two different factors simultaneously: pathological conditions and age. Sphingolipid metabolism is the top most altered pathway in young STEMI patients and as such may represent a valuable prognostic factor and potential therapeutic target.</p>

Diagnostic value of absent ductular reaction at hepatocellular-stromal boundaries in early stage hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of absent ductular reaction (DR) at hepatocellular-stromal boundaries in early stage hepatocellular carcinoma (HCC) with cirrhosis in patients with chronic hepatitis B.</p><p><b>METHODS</b>Cytokeratin (CK)7 and CK19 expression was detected by the SP immunohistochemistry method in 112 hepatic nodules taken from 20 cases of early HCC, 26 cases of HCC with nodules more than 3 cm, 20 cases of high-grade dysplastic nodule (HGDN), 26 cases of low-grade dysplastic nodule (LGDN), and 20 cases of cirrhosis (CIR). DR/CK7 and DR/CK19 were assessed separately on a semi-quantitative scale and statistically analyzed.</p><p><b>RESULTS</b>The mean age of the patients in the study was 53.71 years-old, and the study population consisted of 73 males and 39 females. The follow-up time ranged from 3 to 90 months. Positive CK7 and CK19 staining was detected in the cytoplasm of DR-positive hepatobiliary cells, interlobular bile duct, and a portion of hepatic cells. All of the DR/CK7- and DR/CK19-positive cells were localized around the non-invasive nodules. Specimens with focal or diffuse DR/CK7- and DR/CK19-loss had more robust stromal invasion. Specimens from early HCC cases showed greater DR/CK19 loss than specimens from HGDN cases, LGDN cases and CIR cases (all P less than 0.01). DR/CK7 loss of early HCC was less than HCC with nodules more than 3 cm (P less than 0.05), and more than LGDN cases and CIR cases (both P less than 0.01).The area under the receiver operating characteristic curve of DR/CK7 was very similar to that of DR/CK19 (P more than 0.05). Pearson's correlation analysis indicated that DR/CK7 and DR/CK19 were positively correlated with tumor-free time (P less than 0.01) and negatively correlated with early recurrence time as well as death rate (both P less than 0.01). Furthermore, cases showing DR/CK7 or DR/CK19 loss had lower overall survival rate and tumor-free survival rate (P less than 0.01) and higher early recurrence rate (P less than 0.01).</p><p><b>CONCLUSION</b>DR/CK7 and DR/CK19 immunostaining may help to distinguish non-invasive HGDNs from both minimally-invasive and overtly-invasive HCCs by identifying small foci of invasion and predicting increased risk of invasiveness.</p>

Analysis of the relationship between hepatitis B virus precore and basal core promoter mutations and acute-on-chronic liver failure / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analyze the relationship between hepatitis B virus (HBV) precore (PC) and basal core promoter (BCP) mutations and HBV-related acute-on-chronic liver failure (HB-ACLF).</p><p><b>METHODS</b>Forty-four patients with HB-ACLF and 28 patients with chronic hepatitis B (CHB; used as controls) were enrolled and venous blood samples were collected from all individuals. The PC and BCP gene fragments were amplified by nested PCR. HBV genotype and BCP/PC mutations were determined by direct sequencing and analysis by BioEdit (version 7.0.9.0). Ten of the HB-ACLF patients were selected for follow-up (range: 2-8 weeks), which included once weekly sera collection to determine the relation of mutations and treatment response. Serum levels of HBV DNA were measured by real-time PCR assay, and alanine aminotransferase, total bilirubin, creatinine and albumin were measured by standard biochemical assay and used to determine the MELD score.</p><p><b>RESULTS</b>All 44 HB-ACLF patients were infected with HBV genotype C. In the CHB group, 26 patients were infected with genotype C and two with genotype B. Single mutations (A1762T, G1764A, T1753V, G1896A, and G1899A) and combined mutations (A1762T + G1764A, G1896A + G1899A, T1753V+ A1762T + G1764A, G1896A + G1899A + A1762T + G1764A, and A1762T + G1764A + G1896A) were more frequently detected in HB-ACLF patients than in CHB patients (P less than 0.05). A significantly higher proportion of PC/BCP wild-type sequences was found in patients with CHB than in patients with HB-ACLF (17.9% vs. 2.3%; x² = 5.440, P = 0.020). The proportion of patients carrying both PC and BCP mutations was significantly higher in HB-ACLF patients than in CHB patients (79.6% vs. 39.3%; x² = 12.021, P = 0.001). The proportion of patients carrying only BCP mutation was 42.9% in the CHB group and 20.5% in the HB-ACLF group (x² = 4.157, P = 0.041). No occurrences of only PC mutation were detected in either the CHB or HB-ACLF group. The combined mutations were present in all 10 of the HB-ACLF follow-up patients. Mutations G1899A, T1753V, and A1846T were correlated with disease recovery. Significant decreases in the MELD score were accompanied by decreases in the A1846T mutation.</p><p><b>CONCLUSION</b>Significantly more HB-ACLF patients carried HBV with mutations in the PC and BCP than CHB patients. Moreover, more HB-ACLF patients carried HBV with PC + BCP combined mutations and PC mutation only. The G1899A, T1753C, and A1846T mutations were associated with HB-ACLF response to treatment and improvement in liver function.</p>

Tumor-infiltrating FoxP3+ Tregs are associated with CD34 expression and prognosis of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the correlation between FoxP3+ regulatory T lymphocytes (Tregs) in hepatocellular carcinomas (HCCs) and peritumoral tissues with CD34 expression and patient prognosis.</p><p><b>METHODS</b>Fifty-five sets of patient-matched tumors and peritumoral tissues were obtained during curative resection for HCC. In situ immunohistochemistry was used to assess and comparatively analyze Treg presence and CD34 expression in each specimen set. The relation between quantified Tregs values and various clinicopathologic factors were evaluated by the Spearman Rank Correlation test. Univariate (Log Rank test) and multivariate (Cox Regression model) analyses were used to determine the potential prognostic value of each factor.</p><p><b>RESULTS</b>The average number of intratumoral Tregs was significantly higher than that in corresponding peritumoral tissues (10.8 (range: 4.4 to 19.4) vs. 1.4 (0.6 to 3.2), respectively; P less than 0.01). The presence of intratumoral Tregs correlated with up-regulated CD34 expression (r = 0.279, P less than 0.05). Increased number of intratumoral Tregs were significantly associated with decreased rates of overall survival (OS, P less than 0.05) and disease-free survival (DFS, P less than 0.05), and was identified as an independent prognostic factor (OS, hazard ratio (HR) = 3.310, 95% confidence interval (CI): 1.368-8.007, P less than 0.01; DFS, HR = 2.666, 95% CI: 1.321 to 6.394, P less than 0.01).</p><p><b>CONCLUSION</b>Intratumoral infiltration by Tregs is a marker of poor prognosis in HCC patients.</p>

Studies on the relationship between polymorphism of IL-28B rs8099917 and the outcome of HBV infection / 中华流行病学杂志

Objective To investigate the correlation between IL-28B rs8099917 polymorphism and the outcome of HBV infection.Methods Genotype ofrs8099917(T＞G) in IL-28B locus was determined by TaqMan SNP genotyping from 486 individuals which including 199 chronic HBV carriers (including 100 HBV-induced liver cirrhosis and 99 HBV-related HCC).143 people with selflimited infection and 144 healthy people served as controls.Multivariate analysis was used to assess the effect of IL-28B rs80999 1 7 SNP among all the studied groups.Results Distribution of genotype and allele of the rs8099917 locus were in accordance with Hardy-Weinberg equilibrium in different groups or with the total population.The frequencies of the rs8099917 TT,GT,GG genotypes were 89.3％,10.5％ and 0.2％,and the frequency of allele T and G accounted for 94.5％ and 5.5％,respectively.In respect of genotype or allele frequency,there was no significant differences found among the groups(P＞0.05 ).When comparing with the TT genotype,data from the multinomial logistic analysis showed that the ORs and (95％CI) of TG/GG genotypes were 1.589 (0.735-3.437),1.351 (0.550-3.316) and 1.704 (0.717-4.052),respectively.The genotype frequencies in different groups with different clinical features showed that TG/GG genotypes significantly increased the risk of r-GT Ⅱ( + ) for individuals with HBV-related HCC (X2=17.534,P=0.001 ),with OR as 14.821 (3.227-68.064).It was particularly so for males(X2=14.924,P=0.014),with OR(95％CI) as 45.000(2.772-730.571 ).Conclusion IL-28B rs8099917 SNP had no correlation with the outcome of HBV infection.

Resistance mutation patterns of hepatitis B virus in patients with suboptimal response to adefovir dipivoxil therapy after lamivudine resistance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the resistance mutation patterns of hepatitis B virus(HBV) during adefovir dipivoxil (ADV) monotherapy or combination therapy after lamivudine(LAM) resistance.</p><p><b>METHODS</b>Serum samples from fifteen patients with suboptimal viral response to ADV therapy after LAM resistance were collected. The RT region of HBV P gene was PCR-applied, cloned and sequenced, and the mutation patterns in relation to resistance were analyzed.</p><p><b>RESULTS</b>The ADV resistance mutation patterns of A181T+N236T, A181V, A181T were selected in ADV monotherapy group. The LAM resistance mutation patterns of M204V+L180M, M204V+L180M+L229V, M204I+L80I, M204V+L180M+V207I were detected in the combination therapy group. 20% of clones from three serum samples were detected double resistance to LAM and entecavir (ETV) in the combination therapy group, the resistance patterns were M204I+L80I+T184I (2/10), M204V+L180M+T184S (2/10), and M204V+L180M+G173L+S202G (2/10) respectively. I269L clones were detected in two serum samples from both two groups and P109S clones also detected in the one from monotherapy group.</p><p><b>CONCLUSIONS</b>In the patients with suboptimal viral response to ADV therapy after LAM resistance, the ADV resistance mutation patterns of A181T+N236T, A181V and A181T could easily be selected during ADV monotherapy; while in the patients with combination therapy, the LAM resistance mutation patterns of M204V+L180M, M204V+L180M+L229V, M204I+L80I, and M204V+L180M+V207I were predominant, the ETV resistance mutation T184I/S and S202G could be selected. The mutation patterns of I269L and P109S may impact the responses to ADV therapy in some patients.</p>

Identification of the regions of copy number amplification associated with hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To screen and determine the regions of copy number variation (CNV) associated with hepatocellular carcinoma (HCC) using SNP array and fluorescence quantitative PCR.</p><p><b>METHODS</b>The CNV from HCC cell line TJ3ZX-01 was analyzed using GeneChip Human Mapping 500K SNP array. According to the data obtained by SNP array analysis, four candidate amplification regions were verified in 41 primary HCC samples by fluorescence quantitative PCR.</p><p><b>RESULTS</b>Four regions of copy number amplification at 1q21.2, 1q22 approximately 23.1, 7p22.1 and 22q13.1 were detected by SNP array analysis. The four candidate amplicons occurred in 56.1% (23/41) of HCC samples at 1q21.2; 80.5% (33/41) at 1q22 approximately 23.1; 75.6% (31/41) at 7p22.1 and 31.7% (13/41) at 22q13.1 analyzed with sequence tagged site (STS) markers by quantitative PCR.</p><p><b>CONCLUSION</b>In four candidate amplification regions selected by SNP array analysis and detected by fluorescence quantitative PCR, three amplification regions show increased copy number in more than 50.0% HCC tissues. This result indicates that these amplification regions are associated with pathogenesis of hepatocellular carcinoma.</p>

Induction of dendritic cells from intra-operative lost blood and their effects on CIK against liver cancer cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the practical possibility of inducing dendritic cells (DCs) from mononuclear cells in the lost blood during operation of hepatocellular carcinoma (HCC) patients, and attempted to find a new source of precursor cells for the personalized immunotherapy based on DCs.</p><p><b>METHODS</b>Collected lost blood during hepatectomy from 9 HCC patients and human cord blood from 8 cases of healthy donors undergoing caesarean section. Their mononuclear cells were divided into monocytes and nonadherent lymphocytes. RhGM-CSF and rhIL-4 were administered to induce the monocytes differentiation into DCs, and then loaded with different antigens (lysate antigen of autologous liver cancer cells and cell line SMMC-7721 cells). The lymphocytes were induced into cytokine-induced killer cells (CIK) with IL-2, CD3-Ab, gamma-IFN and PHA. MTT assay was performed to detect the proliferation rate of T lymphocytes mediated by DC and the cytotoxicity of CIK to liver cancer cells.</p><p><b>RESULTS</b>DCs induced from monocytes of the intra-operative lost blood possessed typical morphology and phenotypes. Compared with the DCs from cord blood, the DCs from intra-operative lost blood expressed lower level of surface markers, but both could effectively induce proliferation of CIK and enhance the cytotoxicity of activated CIK against liver cancer cells at similar levels. When the DCs from lost blood and their counterpart from cord blood were both loaded with autologous tumor cell antigen, the proliferation rates of CIK were (388.9 +/- 137.3)% and (315.1 +/- 44.5)%, respectively, and the killing rates against tumor cells were (87.1 +/- 8.0)% and (90.0 +/- 5.1)%, respectively. When the two similar DC groups were loaded with lysate antigen of SMMC-7721 cells, the proliferation rates of CIK were (239.9 +/- 48.7)% and (226.3 +/- 32.3)%, respectively, and the killing rates against tumor cells were (76.4 +/- 7.9)% and (81.1 +/- 4.3)%, respectively. There were no significant differences between those two DC groups. The data also showed that the proliferation and cytotoxicity of CIK induced by DCs loaded with autologous antigen were higher than that of DCs loaded with SMMC-7721 antigen.</p><p><b>CONCLUSION</b>Mononuclear cells separated from intra-operative lost blood of HCC patients can be induced into mature DCs, which can effectively activate CIK and significantly increase its killing effect on the liver cancer cells, and may become a new source of DCs to study and develop vaccines for clinical application.</p>

Aberrant methylation of multiple genes and its clinical implication in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the methylation frequencies of multiple tumor suppressor genes (TSGs) in hepatocellular carcinoma (HCC) and the clinical implication of aberrant DNA methylation in molecular carcinogenesis of HCC.</p><p><b>METHODS</b>Sixty samples of HCC and the paired adjacent liver tissue, 16 samples from post-hepatitis cirrhotic livers, 5 from livers with chronic hepatitis and 5 from normal livers were collected. Eight TSGs frequently silenced by hypermethylation of their promoters in various types of digestive tumors were selected, including APC, RASSF1A, p16, GSTP1, MGMT, DAPK, SOCS-1 and RIZ1. The status of promoter methylation in these 8 genes was investigated using methylation-specific polymerase chain reaction. The clinicopathological data of HCC were also analyzed in order to evaluate the clinical implication of aberrant methylation in HCC.</p><p><b>RESULTS</b>Methylation of the 8 TSGs was quite frequent in HCC, with a methylation rate of 95.0% in RASSF1A, 90.0% in APC, 73.3% in GSTP1, 65.0% in p16, 61.6% in RIZ1 and 60.0% in MGMT. Methylation of the 6 genes was more frequent in HCC than that in adjacent tissues (P < 0.05). The methylation rate of MGMT, GSTP1 and RIZ1 in the adjacent tissues was 41.6%, 40.0% and 25.0%, respectively, significantly higher than that in cirrhotic liver (P < 0.05). p16 methylation was more frequently observed in HCC in elderly patients. The frequency of MGMT methylation was tended to be higher in giant HCC than that in the other types of HCC. Patients with MGMT methylation in the tumor were found to have a shorter disease free survival.</p><p><b>CONCLUSION</b>Different frequency of methylation in hepatocellular carcinomas, adjacent liver tissues and cirrhotic livers implies that epigenetic alteration in the hepatocellular carcinogenesis may be a gradually progressive process. Methylation status of MGMT, GSTP1 and RIZ1 may be promising in risk assessment of hepatocellular carcinoma and in early diagnosis. Furthermore, MGMT methylation might be also used as a potential prognostic biomarker for hepatocellular carcinoma patients.</p>

Construction of anti-lung tumor gene differentially expressed bank of wild mouse / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To construct the anti-lung tumor gene differentially expressed bank of wild mouse and to explore the mechanisms of the TW wild mouse suppressing the occurring of lung tumor.</p><p><b>METHODS</b>Using suppression subtractive hybridization (SSH) technique, the differentially expressed genes between TAF1 mouse and A/wy mouse were selected out and the subtracted cDNA bank was constructed. 166 clones were performed DNA sequencing and then were assayed by blast programme.</p><p><b>RESULTS</b>Among the blast results of 166 differentially expressed clones, 87 known genes (mRNA or cDNA) were in homology with 134 clones and were divided into 7 classifications according to the biological role.14 DNA fragments were in homology with 32 clones, in which 20 clones were in homology with 9 mouse DNA sequences, 2 clones were in homology with one bacterial gene sequences and 3 clones were clone vector.</p><p><b>CONCLUSION</b>With SSH technique, the anti-lung tumor gene differentially expressed bank of wild mouse are successfully constructed.</p>