Combination of serum tumor markers dickkopf-1, DCP and AFP for the diagnosis of primary hepatocellular carcinoma

Objective To evaluate the detection accuracy of the biomarkers dickkopf-1, DCP and AFP as a serum biomarker panel by comparing the sensitivity of the panel with those of the individual biomarkers. Methods The study was composed of three groups, one with HCC patients, one with non-HCC liver diseases and one with healthy controls. Serum AFP was measured using a chemiluminescence assay and serum dickkopf-1 and DCP were measured with ELISA. The sensitivity and specificity of the biomarkers were analyzed as single parameters and as a serum panel. Results The HCC group showed higher levels of dickkopf-1, DCP and AFP than the other two groups (P 0.05). The combination of the biomarkers as a serum panel produced much better sensitivity (93.02%) and specificity (78.00%) than each of the markers individually (P < 0.05). Conclusion The combination of AFP, DCP and dickkopf-1 as a biomarker panel can significantly improve the detection power with much higher sensitivity and specificity for HCC than any of the biomarkers alone. The tests are convenient and inexpensive, and may serve as a valuable addition to current options for the diagnosis of HCC.

Combination of serum tumor markers dickkopf-1, DCP and AFP for the diagnosis of primary hepatocellular carcinoma

OBJECTIVE@#To evaluate the detection accuracy of the biomarkers dickkopf-1, DCP and AFP as a serum biomarker panel by comparing the sensitivity of the panel with those of the individual biomarkers.@*METHODS@#The study was composed of three groups, one with HCC patients, one with non-HCC liver diseases and one with healthy controls. Serum AFP was measured using a chemiluminescence assay and serum dickkopf-1 and DCP were measured with ELISA. The sensitivity and specificity of the biomarkers were analyzed as single parameters and as a serum panel.@*RESULTS@#The HCC group showed higher levels of dickkopf-1, DCP and AFP than the other two groups (P  0.05). The combination of the biomarkers as a serum panel produced much better sensitivity (93.02%) and specificity (78.00%) than each of the markers individually (P < 0.05).@*CONCLUSION@#The combination of AFP, DCP and dickkopf-1 as a biomarker panel can significantly improve the detection power with much higher sensitivity and specificity for HCC than any of the biomarkers alone. The tests are convenient and inexpensive, and may serve as a valuable addition to current options for the diagnosis of HCC.

Coagulase-negative staphylococcus and enterococcus as predominant pathogens in liver transplant recipients with Gram-positive coccal bacteremia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Gram-positive bacteria such as Staphylococcus aureus have been a common cause of infection among liver transplant (LT) recipients in recent decades. The understanding of local epidemiology and its evolving trends with regard to pathogenic spectra and antibiotic susceptibility is beneficial to prophylactic and empiric treatment for LT recipients. This study aimed to investigate etiology, timing, antibiotic susceptibility and risk factors for multidrug resistant (MDR) Gram-positive coccal bacteremia after LT.</p><p><b>METHODS</b>A cohort analysis of prospectively recorded data was performed to investigate etiologies, timing, antibiotic susceptibility and risk factors for MDR Gram-positive coccal bacteremia in 475 LT recipients.</p><p><b>RESULTS</b>In 475 LT recipients in the first six months after LT, there were a total of 98 episodes of bacteremia caused by Gram-positive cocci in 82 (17%) patients. Seventy-five (77%) bacteremic episodes occurred in the first post-LT month. The most frequent Gram-positive cocci were methicillin-resistant coagulase-negative staphylococcus (CoNS, 46 isolates), methicillin-resistant Staphylococcus aureus (MRSA, 13) and enterococcus (34, E. faecium 30, E. faecalis 4). In all Gram-positive bacteremic isolates, 59 of 98 (60%) were MDR. Gram-positive coccal bacteremia and MDR Gram-positive coccal bacteremia predominantly occurred in patients with acute severe exacerbation of chronic hepatitis B and with fulminant/subfulminant hepatitis. Four independent risk factors for development of bacteremia caused by MDR Gram-positive coccus were: LT candidates with encephalopathy grades II - IV (P = 0.013, OR: 16.253, 95%CI: 1.822 - 144.995), pre-LT use of empirical antibiotics (P = 0.018, OR: 1.029, 95%CI: 1.002 - 1.057), post-LT urinary tract infections (P < 0.001, OR: 20.340, 95%CI: 4.135 - 100.048) and abdominal infection (P = 0.004, OR: 2.820, 95%CI: 1.122 - 10.114). The main infectious manifestations were coinfections due to gram-positive cocci and gram-negative bacilli.</p><p><b>CONCLUSIONS</b>Methicillin-resistant CoNS and enterococci are predominant pathogens among LT recipients with Gram-positive coccal bacteremia. Occurrences of Gram-positive coccal bacteremia may be associated with the severity of illness in the perioperative stage.</p>

Spectrum and risk factors for invasive candidiasis and non-Candida fungal infections after liver transplantation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Invasive fungal infections are an important cause of posttransplant mortality in solid-organ recipients. The current trend is that the incidence of invasive candidiasis decreases significantly and invasive aspergillosis occurs later in the liver posttransplant recipients. The understanding of epidemiology and its evolving trends in the particular locality is beneficial to prophylactic and empiric treatment for transplant recipients.</p><p><b>METHODS</b>A retrospective analysis was made of recorded data on the epidemiology, risk factors, and mortality of invasive fungal infections in 352 liver transplant recipients.</p><p><b>RESULTS</b>Forty-two (11.9%) patients suffered from invasive fungal infection. Candida species infections (53.3%) were the most common, followed by Aspergillus species (40.0%). There were 21 patients with a superficial fungal infection. The median time to onset of first invasive fungal infection was 13 days, first invasive Candida infection 9 days, and first invasive Aspergillus infection 21 days. Fifteen deaths were related to invasive fungal infection, 10 to Aspergillus infection, and 5 to Candida infection. Invasive Candida species infections were associated with encephalopathy (P = 0.009) and postoperative bacterial infection (P = 0.0003) as demonstrated by multivariate analysis. Three independent risk factors of invasive Aspergillus infection were posttransplant laparotomy (P = 0.004), renal dysfunction (P = 0.005) and hemodialysis (P = 0.001).</p><p><b>CONCLUSIONS</b>The leading etiologic species of invasive fungal infections are Candida and Aspergillus, which frequently occur in the first posttransplant month. Encephalopathy and postoperative bacterial infection predispose to invasive Candida infection. Posttransplant laparotomy and poor perioperative clinical status contribute to invasive Aspergillus infection. More studies are needed to determine the effect of prophylactic antifungal therapy in high risk patients.</p>

Reduction of endotoxin may protect lung from injury in rats with hepatopulmonary syndrome / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the protective effects of ofloxacin by reduction of endotoxin on lung in rats with hepatopulmonary syndrome (HPS).</p><p><b>METHODS</b>Rat models of HPS were induced by ligating the bile duct to result in the biliary cirrhosis. Thirty male SD rats were randomly divided into three groups with 10 animals in each one: sham operation group (the bile ducts were isolated and 2 ml NS was injected i.p), HPS model group (the bile ducts were ligated and 2 ml NS was injected i.p) and levofloxacin group (the bile ducts were did as the former group and 20 mg/kg ofloxacin injected i.p). After 6 weeks the portal pressure, ratios of dry weight to wet weight of lung (D/W), weight of spleen, plasma levels of endotoxin, MPO active and MDA contents in the lung. Plasma and lavage fluid of lung were examined. The results of the blood-gas analysis, the bacterial culture of blood, bile and ascites and the evaluation of pathologic change of lung be also investigated.</p><p><b>RESULTS</b>In levofloxacin group, portal pressure [(19.28 +/- 2.3) compare with (17.80 +/- 2.18)cm H2O, P<0.01], D/W [(5.1 +/- 0.7) compare with (4.9 +/- 0.7), P <0.01], plasma endotoxin levels [(59 +/- 6.2) compare with (268 +/- 35.6)ng/L, P<0.01], MPO active [(0.40 +/- 0.10) compare with (0.24 +/- 0.03)U, P<0.01] and MDA [(0.32 +/- 0.05) compare with (0.22 +/- 0.03) micromol/L, P<0.01] contents in the lung were significantly decreased compared with those in HPS group, and the inflammation of lung was also obviously alleviated.</p><p><b>CONCLUSION</b>Reduction of endotoxin can attenuate the injury of lung in HPS rats.</p>

Efficacy of hepatitis B immunoglobulin in relation to the gene polymorphisms of human leukocyte Fcgamma receptor III (CD16) in Chinese liver transplant patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Although the use of hepatitis B immunoglobulin (HBIG) may lead to a significant reduction in recurrent hepatitis B virus (HBV) infection and improve the survival of patients who have undergone liver transplantation (LT) for hepatitis B-related diseases, the recurrence of the disease still remains at a lower level. Different clinical curative effects were observed in patients with the same HBV-related diseases and the same therapy. This study was undertaken to investigate whether the efficacy of HBIG is associated with FCGR3A gene polymorphisms in Chinese liver transplant patients.</p><p><b>METHODS</b>Altogether 77 patients who had received liver transplantation for hepatitis B-related diseases with more than one-year survival after surgery were studied. The recurrence of HBV was characterized by the appearance of HBsAg in serum after the operation. The FCGR3A genotyping was performed using genomic DNA sequencing (ABI 3037). Single nucleotide polymorphism at nucleotide 559 was detected by Polyphred.</p><p><b>RESULTS</b>Of the 77 patients, 14 were complicated with HBV recurrence post-transplant. The FCGR3A at nucleotide 559 TT was observed in 35 (45.5%) subjects, whereas TG in 31 (40.3%) and GG in 11 (14.3%). In the 559G carrier group (n = 42, 54.5%), the risk of HBV recurrence was 9.5%, and 1- and 2-year recurrence-free survival rates were 95.2% and 88.7%, respectively. In the 559G noncarrier group (n = 35, 45.5%), the risk of HBV recurrence was 28.6%, and 1- and 2-year recurrence-free survival rates were 74.3% and 69.3%, respectively. The risk of HBV recurrence and the recurrence-free survival rate were both statistically different between the 559G carrier and noncarrier groups (P < 0.05).</p><p><b>CONCLUSIONS</b>A single nucleotide polymorphism (T/G) at position 559 of the FCGR3A gene was found in Chinese patients. The efficacy of HBIG in prophylaxis of HBV recurrence after LT is associated with the gene polymorphism, so detecting FCGR3A genotypes can be a clinical reference of the HBIG administration.</p>

Infusion of nonmyeloablative bone marrow alleviates acute rejection reaction in liver allotransplantation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To study the effect and implication of nonmyeloablative donor specific bone marrow (DSBM) infusion on the immunoreaction of liver allotransplantation.</p><p><b>METHODS</b>Orthotopic liver transplantation model was used in this study. Groups were set as follows: Group I, syngeneic control (Wistar-to-Wistar); Group II, acute rejection (SD-to-Wistar); Group III, acute rejection treated with cyclosporine A (CsA) by intramuscular injection (SD-to-Wistar+CsA); Group IV, bone marrow infusion at 7 d pretransplantation followed by short-term CsA treatment (SD-to-Wistar+DSBM); Another group of short-term CsA treatment preoperatively without bone marrow infusion was also set as control. General characteristics and survival time were observed. Histological grades of rejection were determined by pathological examination. IL-2 and IFN-gamma level in peripheral blood and donor liver were detected respectively by Enzyme-Linked Immuno-Sorbent Assay (ELISA) and Western blot. Chimerism of donor cells was measured by PCR for a male-specific marker (Y-chromosome-specific sequence, Sry).</p><p><b>RESULTS</b>No signs of rejection were found in Group I. Acute rejection occurred in both Group II and the short-term CsA treated group. All the recipients died at (9-15) d posttransplantation with a median survival time of (10.7+/-0.5) d and (11.2+/-2.4) d, respectively. Only mild rejection could be seen in Group III. In Group IV, 4 out of 6 recipients had long-term survival (>100 d), the histological grade of rejection was significantly lower than that of Group II, so did the expression level of IL-2 and IFN-gamma in both peripheral blood and grafted liver. Y-chromosome-specific sequence (Sry) of male SD rats could be detected in the bone marrow, spleen and thymus of female recipients at 15 d after bone marrow infusion.</p><p><b>CONCLUSION</b>Mild preconditioning nonmyeloablative donor specific bone marrow infusion can enhance chimerism formation in recipients, alleviate the rejection of liver allotransplantation and prolong survival of liver allotransplantation.</p>

Role of 1,25-dihydroxyvitamin D3 in preventing acute rejection of allograft following rat orthotopic liver transplantation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We investigated the role of 1,25-dihydroxyvitamin D3(1,25-(OH)2D3) in preventing allograft from acute rejection following orthotopic liver transplantation.</p><p><b>METHODS</b>A rat orthotopic liver transplantation model was used in this study. SD-Wistar rats served as a high responder strain combination. Recipients were subjected to administration of 1,25-(OH)2D3 at dosages ranging from 0.25 microg.kg(-1).d(-1) to 2.5 microg.kg(-1).d(-1). Survival after transplantation as well as pathological rejection grades and IFN-gamma mRNA, IL-10 mRNA transcription intragraft on day 7, and day 30 post-transplantation were observed.</p><p><b>RESULTS</b>After recipients were treated with 1,25(OH)2D3 at dosages of 0.5 microg.kg(-1).d(-1) or 1.0 microg.kg(-1).d(-1), survivals of recipients were prolonged. Ninety-five percent confidence intervals of survival were 46 - 87 days and 69 - 102 days (both P = 0.0005 vs control group), respectively. On day seven post-transplantation, relative levels of IFN-gamma mRNA transcription were 0.59 +/- 0.12 and 0.49 +/- 0.16, which was higher than the control group (P = 0.005, P = 0.003, respectively). Relative levels of IL-10 mRNA transcription were 0.83 +/- 0.09 and 0.76 +/- 0.09, which was lower than the control group (P = 0.002, P = 0.003, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and on day thirty post-transplantation were 1.5 and 2.0 in comparison with the CsA-treated group (P = 0.178, P = 0.171, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and day thirty post-transplantation were 1.5 and 1.5 in comparison with CsA-treated group (P = 0.350, P = 0.693, respectively).</p><p><b>CONCLUSION</b>After each recipient was treated with 1,25-(OH)2D3 at a dosage of (0.5 - 1.0) microg.kg(-1).d(-1), transcription of cytokine intragraft was accommodated effectively and deviated to Th2 type, resulting in alleviation of acute rejection. 1,25-(OH)2D3 can prolong survival of recipient after orthotopic liver transplantation.</p>

Ciclosporin down-regulates interferon-gamma gene transcription via its inhibition of nuclear factor kappaB activity after liver transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the relation between the activity of nuclear factor kappaB (NF-kappaB) and the expression of interferon (IFN)-gamma gene transcription with or without ciclosporin treatment after liver transplantation.</p><p><b>METHODS</b>Orthotopic liver transplantation was performed in this study. Group I: syngeneic control (Wistar-to-Wistar); Group II: acute rejection (SD-to-Wistar); Group III: acute rejection treated with ciclosporin by intramuscular route (SD-to-Wistar + ciclosporin). Electrophoretic mobility shift assay and reverse transcriptase polymerase chain reaction were used to analyze NF-kappaB activity of splenocytes and IFN-gamma gene transcription expression of grafted liver with or without ciclosporin treatment after liver transplantation. Histopathological examination was also used in this study.</p><p><b>RESULTS</b>Low NF-kappaB activity was only detected at day 5 and day 7 in Wistar-to-Wistar group after transplantation, meanwhile low IFN-gamma mRNA expression was detected at any time in this group. In contrast, high NF-kappaB activity was detected in SD-to-Wistar group and high level IFN-gammamRNA expression was detected at all time points in this group. The activity of NF-kappaB and IFN-gammamRNA expression were significantly inhibited in SD-to-Wistar + ciclosporin group which was significantly lower than that of SD-to-Wistar group (P < 0.001). A good correlation was found between activity of NF-kappaB and IFN-gamma mRNA expression in this study (r=0.815, P <0.01).</p><p><b>CONCLUSIONS</b>The change of expression IFN-gamma mRNA is at least partially due to the activity change of NF-kappaB after orthotopic liver transplantation. CsA down-regulates NF-kappaB activity and further inhibit IFN-gamma gene transcription.</p>

Serum cytokine profiles in stable survivors with clinical liver transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>To elucidate the profile of serum cytokines and adhesion molecules in stable survivors with clinical liver transplantation.</p><p><b>METHODS</b>Flow cytometric analysis was used to analyse the phenotype of T cell subsets in peripheral blood mononuclear cells (PBMCs) from group of liver transplantation (LTx) (n = 22), primary liver carcinoma (PLC) (n = 13) and healthy control (n = 12). Enzyme-linked immunoabsorbent assay (ELISA) was used to determine the serum cytokines and adhesion molecules profiles in stable survivors with clinical liver transplantation.</p><p><b>RESULTS</b>Percentage of CD3(+) T cell and CD8(+) T cell, as well as ratio of CD4(+) to CD8(+) revealed no difference among three groups. The percentage of CD3(+)CD25(+) T cells in LTx group was found higher than that in healthy group (P = 0.022). Th1 cytokines (IL-2, IFN-gamma) and Th2 cytokines (IL-4, IL-10), as well as TNF-alpha displayed no significant difference among three groups. The levels of IL-6, ICAM-1 and P-selectin in serum were not found any difference between LTx group and PLC group, while the levels of IL-6, ICAM-1 and P-selectin in serum shown significant difference between LTx and healthy groups (P = 0.048, 0.000 and 0.025, respectively).</p><p><b>CONCLUSIONS</b>Our data demonstrates that effector T-cells can also be activated and exert immunoresponse to grafts permanently under the treatment of immunosuppressant. Adhesion molecules (ICAM-1, P-Selectin) and pro-inflammatory cytokines (IL-6, TNF-alpha) might be involved in the process of chronic graft damage induced by allo-immunoresponse.</p>

The effect of pro-angiogenic factors and their receptors on angiogenesis in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of pro-angiogenic factors and their receptors on angiogenesis in hepatocellular carcinoma.</p><p><b>METHODS</b>Expression of VEGF/KDR and Angiopoietins/Tie2 was detected by RT-PCR and Western blot in 15 cases with hepatocellular carcinoma, 15 tumor adjacent tissues (<1 cm, >5 cm), 8 cirrhotic liver, and 4 normal liver. Immunohistochemistry (IHC) was used to detect CD34 expression, and the relationship between neovascular density and angiogenesis was analyzed.</p><p><b>RESULTS</b>The expression levels of VEGF and Ang2 were significantly higher in hepacellular carcinoma group than those in the other groups (P < 0.01), and so did the expression of CD34. The expressions of KDR and Ang1/Tie2 showed no significant difference in all groups, but they indeed increased to various levels in tumor and tumor adjacent tissues as compared with those in cirrhosis and normal liver.</p><p><b>CONCLUSION</b>VEGF/KDR and Angiopoietins/Tie2 may be the crucial signal pathways in the development of hepatocellular carcinoma.</p>