Evaluation of diagnostic efficacy of NRP-1/CD304 in hematological diseases.

BACKGROUND: Previous studies had explored the diagnostic or prognostic value of NRP-1/CD304 in blastic plasmacytoid dendritic cell neoplasm (BPDCN), acute myeloid leukemia (AML), and B-cell acute lymphoblastic leukemia (B-ALL), whereas the expression and application value of NRP-1/CD304 in other common hematological diseases have not been reported. METHODS: Bone marrow samples from 297 newly diagnosed patients with various hematological diseases were collected to detect the expression of NRP-1/CD304 by flow cytometry (FCM). The diagnostic efficacy of NRP-1/ CD304-positive diseases was analyzed by receiver operating characteristic (ROC) curve, and the area under the ROC curve (AUC) was compared. RESULTS: In the research cohort, the total positive rate of NRP-1/CD304 was 14.81% (44/297), mainly distributed in BPDCN (100%, 6/6), B-ALL (48.61%, 35/72), and AML (4.48%, 3/67), with statistically significant differences (p < 0.01). Other diseases, such as T-cell acute lymphoblastic leukemia (T-ALL), B-cell non-Hodgkin lymphoma (B-NHL), T/NK-cell lymphoma and plasma cell neoplasms, did not express NRP-1/CD304. The AUC of NRP-1/CD304 was 0.936 (95% CI 0.898-0.973), 0.723 (95% CI 0.646-0.801), and 0.435 (95% CI 0.435) in BPDCN, B-ALL and AML, respectively. Besides, CD304 was commonly expressed in B-ALL with BCR-ABL1 gene rearrangement (p = 0.000), and CD304 expression was positively correlated with CD34 co-expression (p = 0.009) and CD10 co-expression (p = 0.007). CONCLUSIONS: NRP-1/CD304 is only expressed in BPDCN, B-ALL and AML, but not in other common hematological diseases. This indicates that NRP-1/CD304 has no obvious diagnostic and follow-up study value in hematological diseases other than BPDCN, B-ALL, and AML.

An Immune Risk Score Predicts Survival of Patients with Acute Myeloid Leukemia Receiving Chemotherapy.

PURPOSE: Prediction models for acute myeloid leukemia (AML) are useful, but have considerable inaccuracy and imprecision. No current model includes covariates related to immune cells in the AML microenvironment. Here, an immune risk score was explored to predict the survival of patients with AML. EXPERIMENTAL DESIGN: We evaluated the predictive accuracy of several in silico algorithms for immune composition in AML based on a reference of multi-parameter flow cytometry. CIBERSORTx was chosen to enumerate immune cells from public datasets and develop an immune risk score for survival in a training cohort using least absolute shrinkage and selection operator Cox regression model. RESULTS: Six flow cytometry-validated immune cell features were informative. The model had high predictive accuracy in the training and four external validation cohorts. Subjects in the training cohort with low scores had prolonged survival compared with subjects with high scores, with 5-year survival rates of 46% versus 19% (P < 0.001). Parallel survival rates in validation cohorts-1, -2, -3, and -4 were 46% versus 6% (P < 0.001), 44% versus 18% (P = 0.041), 44% versus 24% (P = 0.004), and 62% versus 32% (P < 0.001). Gene set enrichment analysis indicated significant enrichment of immune relation pathways in the low-score cohort. In multivariable analyses, high-risk score independently predicted shorter survival with HRs of 1.45 (P = 0.005), 2.12 (P = 0.004), 2.02 (P = 0.034), 1.66 (P = 0.019), and 1.59 (P = 0.001) in the training and validation cohorts, respectively. CONCLUSIONS: Our immune risk score complements current AML prediction models.

Rinsing sampling of core needle biopsy for flow cytometric analysis: A favorable method for lymphoma diagnosis.

BACKGROUND: Conventional protocols utilize core needle biopsy (CNB) or fine needle aspiration (FNA) to produce cell suspension for flow cytometry (FCM) is a diagnostic challenge for lymphoid malignancies. We aim to develop an alternative CNB rinsing technique (RT) to produce cell suspension for FCM during this mini-invasive procedure of CNB for lymphoma diagnosis. METHODS: FNA and CNB specimens from the same lesion of 93 patients with suspected lymphoma were collected under the guidance of B-ultrasound simultaneously. The fresh CNB samples were prepared to cell suspension by RT for FCM immunophenotyping analysis (Group CNB-RT). Then, the CNB tissues after performing the RT process and the fresh FNA tissues were processed by conventional tissue cell suspension (TCS) technique to obtain the cell suspensions (Groups of CNB-TCS & FNA-TCS), respectively, as comparison. The diagnostic efficacies, as well as the concordances of the FCM results with reference to the morphologic diagnoses were compared in these three groups. RESULTS: RT could yield sufficient cells for FCM immunophenotyping analysis, though a lower cell numbers compared to TCS technique. The diagnostic concordance was comparable in group CNB-RT (91.1%) to the group CNB-TCS (88.9%) and group FNA-TCS (88.4%) (p = 0.819). The diagnostic sensitivity and specificity of CNB-RT (91.1%; 100%) was not inferior to that of CNB-TCS (88.9%; 100%) and FNA-TCS (88.4%; 98.8%). CONCLUSIONS: This study shows the CNB-RT presented non-inferior diagnostic concordance and efficacy as compared to the TCS technique. CNB-RT has the potential to produce cell suspension for FCM immunophenotyping while preserving tissue for lymphoma diagnosis and research.

A novel score based on serum apolipoprotein A-1 and C-reactive protein is a prognostic biomarker in hepatocellular carcinoma patients.

BACKGROUND: The aim of this study was to propose a prognostic scoring system based on preoperative serum apolipoprotein A-1 and C-reactive protein (ApoA-1 and CRP, AC score) levels and to evaluate the prognostic value of these markers in patients with hepatocellular carcinoma (HCC). METHODS: In all, 539 consecutive cases diagnosed with HCC from 2009 to 2012 at Sun Yat-sen University Cancer Center were analysed. The characteristics and levels of pretreatment lipids (ApoA-1, apolipoprotein B (Apo-B), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglycerides (TGs)) and CRP were reviewed and determined by univariate and multivariate Cox hazard models. Then, the AC score was proposed, which combines two independent risk factors (ApoA-1 and CRP). RESULTS: The optimal cut-off points in our study were determined according to established reference ranges. Patients with decreased ApoA-1 levels (< 1.090 g/L) and increased CRP levels (≥3.00 mg/L) exhibited a significantly poor overall survival (OS) and disease-free survival (DFS). The AC score was calculated as follows: patients with decreased ApoA-1 and elevated CRP were given a score of 3, patients with only one of these abnormalities were given a score of 2, and those with no abnormalities were given a score of 1. Patients with a higher AC score showed more progressive disease and a poorer prognosis. This was observed not only in the entire cohort (for OS, P < 0.001; for DFS, P < 0.001) but also in the subgroups stratified by pathological stage (stage I-II and stage III-IV). The discriminatory ability of the AC score in HCC was assessed according to the AUC values. The AUC value of the AC score (AUC: 0.676, 95% CI: 0.629-0.723, P < 0.001) was higher than that of AFP. In addition, the combination of the AFP and AC scores (AUC: 0.700, 95% CI: 0.655-0.745, P < 0.001) was superior to the AFP and AC scores alone. CONCLUSIONS: The AC score is a significant valuable predictor of OS and DFS and could more accurately differentiate the prognosis of HCC patients. As this study is a retrospective analysis, the value of the AC score should be validated in large prospective trials.

A Simple and Effective Method for Flow Cytometric Study of Lymphoid Malignancies Using Needle Core Biopsy Specimens.

OBJECTIVES: We developed a simple and effective rinsing technique (RT) of needle biopsies to produce cell suspensions for flow cytometry (FCM) and evaluated whether the RT is comparable to the conventional tissue cell suspension (TCS) technique. METHODS: We retrieved 93 needle core biopsy cases employing the RT for FCM and 25 needle biopsy cases using TCS for FCM. RESULTS: The diagnostic concordance between the FCM results and the morphologic diagnoses of both groups was compared. The diagnostic concordance was comparable in the RT group (92.6%) to the TCS group (71.4%). Furthermore, the diagnostic concordance in the RT group was associated with number of isolated cells. The diagnostic accuracy increased significantly when the cell number was above 30,000 in the RT group. CONCLUSIONS: The RT for FCM not only maximizes the tissue utilization, but also is a simple and effective method to obtain cell suspension as compared to traditional cell suspension technique. © 2018 International Clinical Cytometry Society.

Chemokine CCL27 is a novel plasma biomarker for identification the nasopharyngeal carcinoma patients from the Epstein-Barr virus capsid antigen-specific IgA seropositive population.

BACKGROUND: To investigate the predictive value of chemokine CCL27 for identifying early stage nasopharyngeal carcinoma (NPC) patients within a population seropositive for Epstein-Barr virus (EBV) capsid antigen-specific IgA (VCA-IgA). METHODS: CCL27 in plasma samples from 104 NPC patients, 112 VCA-IgA-positive healthy donors, and 140 VCA-IgA-negative normal subjects was measured by ELISA. Expression of CCL27 in nasopharyngeal tissue from 20 VCA-IgA-positive healthy donors and 20 NPC patients was examined by immunohistochemical staining. RESULTS: Levels of CCL27 in the plasma of VCA-IgA-positive healthy donors (607.33 ± 218.81 pg/ml) were significantly higher than the levels in all NPC patients (437.09 ± 217.74, P = < 0.0001) and in the subset of patients with early stage NPC (463.85 ± 226.17, P = 0.0126). Plasma CCL27 levels were significantly lower in the VCA-IgA-negative normal subjects (358.22 ± 133.15 pg/ml) than in either the VCA-IgA-positive healthy donors (P < 0.0001) or the NPC patients (P = 0.0113). CCL27 protein was detected in 16 of 20 (80%) nasopharyngeal tissue samples from VCA-IgA-positive healthy donors and in 3 of 20 (15%) tumor tissue samples from NPC patients. There was no relationship between CCL27 levels and VCA-IgA titers or plasma EBV DNA content. Receiver operating characteristic (ROC) curves demonstrated that plasma CCL27 levels had a sensitivity of 67.00%, a specificity of 73.10%, and an area under the ROC of 0.725 (95% confidence interval [CI]: 0.657-0.793) for distinguishing between NPC patients and VCA-IgA-positive healthy donors. Further analysis showed that CCL27 levels could distinguish between early stage NPC patients and VCA-IgA-positive healthy donors with an area under the ROC of 0.712 (95% CI: 0.560-0.865), a sensitivity of 59.80%, and a specificity of 84.60%. CONCLUSIONS: Chemokine CCL27 could successfully identify NPC patients within a VCA-IgA-positive population.

A retrospective discussion of the prognostic value of combining prothrombin time(PT) and fibrinogen(Fbg) in patients with Hepatocellular carcinoma.

Aims: The levels of coagulation system tests have been studied in various cancers. In this study, our aim is to evaluate the prognostic value of pretreatment plasma coagulation tests in hepatocellular carcinoma (HCC) patients. Patient and methods: A retrospective study was performed in 539 patients with HCC, and follow-up period was at least 60 months until recurrence or death. The prognostic significance of coagulation system tests (prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen) were determined by univariate and multivariate Cox hazard models. Then, according to the results of the multivariate analyses, we proposed the coagulation-Based Stage, which combined the independent risk factors (prothrombin time and fibrinogen). Results: Coagulation system tests including prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fbg) were analyzed. Patients with prolonged PT (≥12.1 sec) levels had significantly poor overall survival (OS) and disease-free survival (DFS), not only in the entire cohort (HR: 1.661, 95%CI: 1.125-2.451, p=0.011 vs. HR: 1.660, 95%CI: 1.125-2.451, p=0.011), but also in the subgroups stratified by pathological stage (stage I-II and stage III-IV). Additionally, high Fbg (≥2.83 g/L) levels experienced significantly decreased OS and DFS (HR: 2.158, 95%CI: 1.427-3.263, p<0.001 vs. HR: 2.161, 95%CI: 1.429-3.267, p<0.001), not only in the entire cohort but also in the subgroups stratified by pathological stage (stage I-II and stage III-IV). All the patients were then stratified (based on combined PT and Fbg) into three groups, The OS for HCC patients were (41.37±17.76), (31.83±19.84) and (18.68±18.41) months, and the DFS for HCC patients were (41.15±17.88), (31.65±19.81) and (18.66±18.39) months. Conclusions: Our findings suggest that the combination of plasma PT and Fbg levels should be evaluated as the valuable predictor of survival in patients with HCC.

Evaluating the diagnostic and prognostic value of circulating cathepsin S in gastric cancer.

To evaluate whether serum Cathepsin S (Cat S) could serve as a biomarker for the diagnosis and prognosis of gastric cancer (GC), Enzyme-linked immuno sorbent assay (ELISA) was used to detect serum Cat S in 496 participants including healthy controls and patients with benign gastric diseases, gastric cancer, esophageal cancer, liver cancer, colorectal cancer, nasopharyngeal cancer and lung cancer. The levels of serum Cat S were significantly increased in cancer patients, especially in GC patients. The qRT-PCR, Western blotting, and immunohistochemical staining revealed the overexpression of Cat S in GC cell lines and tissues. The diagnostic value of serum Cat S for GC patients from controls resulted in an AUC of 0.803 with a sensitivity of 60.7% and a specificity of 90.0%. Moreover, the levels of serum Cat S were associated with GC tumor volume, lymphoid nodal status, metastasis status, and stages. Moreover, the patients with high levels of serum Cat S had a poorer overall survival. Univariate analysis revealed Cat S expression was a prognostic factor. The knockdown of Cat S significantly suppressed the migration and invasion of GC cells. This study suggested serum Cat S may be a potential biomarker for the diagnosis and prognosis of GC.

High level of interleukin-10 in serum predicts poor prognosis in multiple myeloma.

BACKGROUND: Interleukin-10 (IL-10) is a inhibiting inflammatory cytokine that plays an important role in immune suppressive microenvironment in multiple myeloma (MM). Whether the level of serum IL-10 could predict treatment response and survival outcomes or not needs to be investigated in MM patients. METHODS: The level of IL-10 in serum was measured using enzyme-linked immunosorbent assay in 188 patients with newly diagnosed MM. RESULTS: The best cutoff value for IL-10 in predicting survival is 169.69 pg ml(-1) with an area under the curve (AUC) value of 0.747 (P<0.001). In all, 92 patients (48.9%) were classified as high-IL-10 group (>169.96 pg ml(-1)) and 96 patients (51.1%) as low-IL-10 group (⩽169.96 pg ml(-1)). The overall response rate (ORR) was 79.2% in low-IL-10 group, significantly higher than that in high-IL-10 group (53.3%, P<0.001). Patients in low-IL-10 group had significantly better survival compared with those in high-IL-10 group (3-year PFS rate: 69.3% vs 13.3%, P<0.001; 3-year OS rate: 93.6% vs 51.9%, P<0.001). Multivariate analysis revealed that serum IL-10 level >169.96 pg ml(-1) at diagnosis and certain cytogenetic abnormalities were two adverse factors for PFS and OS. CONCLUSIONS: Our data suggest that serum IL-10 at diagnosis is a novel, powerful predictor of prognosis for MM.

Preoperative D-dimers as an independent prognostic marker in cervical carcinoma.

Activation of coagulation and fibrinolysis has been observed in many tumors. Our study aimed to investigate the clinical and prognostic significance of various plasma coagulation tests in patients with cervical cancer. A total of 296 patients with cervical cancer were included in the analysis. Patients were followed up for at least 60 months until death. Pretreatment parameters including activated partial thromboplastin time, D-dimer, fibrinogen, prothrombin time, thrombin time, lactate dehydrogenase, and squamous cell carcinoma antigen were evaluated. Prothrombin time (hazard ratio = 1.825; P = 0.006) and plasma D-dimer levels (hazard ratio = 2.179; P = 0.036) were identified as significant independent predictors of overall survival. Patients with elevated D-dimer levels had a significantly shorter overall survival compared with those with low-D-dimer levels (<0.5 µg/ml) in the stage I subgroup (n = 98, P = 0.019) and stage II subgroup (n = 77, P = 0.044). D-dimer levels differed significantly according to mortality (P < 0.001), stage I versus stage II (P = 0.030), and stage I versus stage III/IV (P = 0.038). DD level of patients with chemotherapy and/or radiotherapy was higher than patients with other treatment (P < 0.001). Patients with a low-D-dimer level (<0.5 µg/ml) showed a significantly better 5-year overall survival (OS) compared with patients with an increased D-dimer level for different histological typing of squamous cell carcinoma (SCC) (P = 0.001) and non-SCC (P < 0.043). In conclusion, the pretreatment plasma D-dimer level is a potential prognostic factor for cervical cancer.

Establishment of using serum YKL-40 and SCCA in combination for the diagnosis of patients with esophageal squamous cell carcinoma.

BACKGROUND: Elevated serum YKL-40 levels have been observed in various cancers. We evaluated the diagnostic performance of serum YKL-40 alone or in combination with the CEA, CYFRA21-1 and SCCA tumor markers for patients with esophageal squamous cell carcinoma (ESCC). METHODS: YKL-40 was detected in ESCC cell lines and tissues by real-time RT-PCR, Western blotting and ELISA. YKL-40 protein expression was determined in 20 ESCC tumor tissues using immunohistochemistry. Serum YKL-40 was measured by ELISA in 126 healthy donors, 59 patients with benign esophageal diseases and 150 patients with ESCC. Serum CEA, CYFRA21-1 and SCCA were determined by electrochemiluminescence. RESULTS: YKL-40 mRNA and protein were observed in ESCC cancer cell lines, tissues and cell culture media, respectively. YKL-40 expression was observed in 17 of 20 ESCC samples (85%). Serum YKL-40 concentration was significantly elevated in patients with ESCC (Range: 6.95-502.10 ng/ml) compared with patients with benign diseases (Range: 1.21-429.30 ng/ml; P = 0.038) and healthy controls (Range: 2.56-132.26 ng/ml; P < 0.001). ROC curves demonstrated that serum YKL-40 has a sensitivity of 72.70%, a specificity of 84.13% and an AUC of 0.874 for the diagnosis of ESCC, which was superior to CEA (Sen: 8.00%; Spe: 96.80%, AUC = 0.652), CYFRA21-1 (Sen: 40.00%; Spe: 92.06%, AUC = 0.746) and SCCA (Sen: 32.67%; Spe: 94.44%, AUC = 0.789). The YKL-40 and SCCA combination was better for diagnosing ESCC (Sen: 82.00%, Spe: 79.37%, PPV: 82.55 and NPV: 78.74; AUC = 0.917) than the YKL-40 and CEA combination (Sen: 74.00%, Spe: 83.20%, PPV: 84.09 and NPV: 72.73; AUC = 0.877), the YKL-40 and CYFRA21-1 combination (Sen: 82.00%, Spe: 77.78%, PPV: 81.46% and NPV: 78.40%; AUC = 0.897) or the CEA, CYFRA21-1 and SCCA combination (Sen: 56.67%, Spe: 84.80%, PPV: 81.73 and NPV: 61.99; AUC = 0.831). Associations between serum YKL-40 levels and the clinic characteristics of ESCC were not significant, with the exception of age (p = 0.001). CONCLUSIONS: ESCC tumor cells and tissues express YKL-40. Serum YKL-40 may be a potential biomarker for ESCC. Serum YKL-40 in combination with SCCA significantly increases the sensitivity of detecting ESCC.

Epstein-Barr virus infection induces indoleamine 2,3-dioxygenase expression in human monocyte-derived macrophages through p38/mitogen-activated protein kinase and NF-κB pathways: impairment in T cell functions.

UNLABELLED: Epstein-Barr virus (EBV) infection has been observed in tumor-infiltrated macrophages, but its infection effects on macrophage immune functions are poorly understood. Here, we showed that some macrophages in the tumor stroma of nasopharyngeal carcinoma (NPC) tissue expressed the immunosuppressive protein indoleamine 2,3-dioxygenase (IDO) more strongly than did tumor cells. EBV infection induced mRNA, protein, and enzymatic activity of IDO in human monocyte-derived macrophages (MDMs). Infection increased the production of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), whereas the neutralizing antibodies against TNF-α and IL-6 inhibited IDO induction. EBV infection also activated the mitogen-activated protein kinase (MAPK) p38 and NF-κB, and the inhibition of these two pathways with SB202190 and SN50 almost abrogated TNF-α and IL-6 production and inhibited IDO production. Moreover, the activation of IDO in response to EBV infection of MDMs suppressed the proliferation of T cells and impaired the cytotoxic activity of CD8(+) T cells, whereas the inhibition of IDO activity with 1-methyl-l-tryptophan (1-MT) did not affect T cell proliferation and function. These findings indicate that EBV-induced IDO expression in MDMs is substantially mediated by IL-6- and TNF-α-dependent mechanisms via the p38/MAPK and NF-κB pathways, suggesting that a possible role of EBV-mediated IDO expression in tumor stroma of NPC may be to create a microenvironment of suppressed T cell immune responses. IMPORTANCE: CD8(+) cytotoxic T lymphocytes (CTLs) play an important role in the control of viral infections and destroy tumor cells. Activation of the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO) in cancer tissues facilitates immune escape by the impairment of CTL functions. IDO expression was observed in some macrophages of the tumor stroma of nasopharyngeal carcinoma (NPC) tissue, and IDO could be induced in Epstein-Barr virus (EBV)-infected human monocyte-derived macrophages (MDMs). NPC cells and macrophages have been found to produce IDO in a gamma interferon (IFN-γ)-dependent manner. Instead, EBV-induced IDO expression in MDMs is substantially mediated by IL-6- and TNF-α-dependent mechanisms via the p38/MAPK and NF-κB pathways, which suppressed the proliferation of T cells and impaired the cytotoxic activity of CD8(+) T cells. This finding provides a new interpretation of the mechanism of immune escape of EBV and shows the immunosuppressive role of EBV-mediated IDO expression in tumor stroma of NPC.

High-density lipoprotein cholesterol is a favorable prognostic factor and negatively correlated with C-reactive protein level in non-small cell lung carcinoma.

BACKGROUND: Although the alterations of lipid profile in lung cancer have been documented, the prognostic value of serum HDL-C level and its correlation with inflammation in NSCLC remain unknown. SUBJECTS AND METHODS: Levels of preoperative serum lipid concentrations (including HDL-C, LDL-C, TC, and TG) and the inflammatory biomarker C-reactive protein level (CRP) were retrospectively analyzed in 228 patients with NSCLC and in 300 healthy controls. The serum lipid levels in these two populations were compared. Univariate and multivariate cox hazards analyses were performed to investigate the prognostic value of serum lipid levels in NSCLC. The correlation between CRP and lipid profile were also analyzed. RESULTS: Compared with those in normal controls, the serum HDL-C, LDL-C, and TC levels were statistically decreased and the TG levels were significantly increased in 228 NSCLC patients. The patients with decreased levels of HDL-C had significantly lower 5-year survival rates than those with normal HDL-C, not only in the whole NSCLC cohort but also in the subgroups stratified according to the disease T, N classifications, and metastasis, whereas the other lipid components were not independent prognostic factors for NSCLC. Of the lipid components, a lower HDL-C level was observed more often in patients with a high CRP level than in those with a normal CRP level. Spearman's rank correlation analysis revealed that the HDL-C level presented a negative correlation with the CRP level (r = -0.360, p<0.001). CONCLUSIONS: A decreased level of preoperative HDL-C was found to be associated with poor survival in patients with NSCLC. Serum HDL-C level may be a clinical prognosis factor for NSCLC patients. In addition, a negative correlation was present between the levels of HDL-C and CRP, the well-known inflammation biomarker.

Low level of high-density lipoprotein cholesterol correlates with poor prognosis in extranodal natural killer/T cell lymphoma.

Studies have found that lymphoma patients often exhibit abnormal lipid metabolism, and a decrease in serum high-density lipoprotein cholesterol (HDL-C) may occur during lymphomagenesis and tumor growth. However, no literatures have investigated the role of HDL-C in patients with extranodal natural killer/T cell lymphoma (ENKTL). In this study, we retrospectively reviewed the HDL-C level in 107 patients newly diagnosed with ENKTL that received either l-asparaginase-based regimen or EPOCH regimen as induction chemotherapy, and evaluated its prognostic value. The mean level of HDL-C was 1.10 mmol/L (range, 0.15­2.63), and the HDL-C level was significantly lower in patients with elevated LDH and beta 2-microglobulin (ß2-MG) (p = 0.017 and 0.001, respectively) and those who underwent disease progression and died (p = 0.031 and 0.007, respectively). In univariate survival analysis, higher Eastern Cooperative Oncology Group performance status score (≥1), Ann Arbor stage III­IV, elevated LDH, higher international prognostic index (IPI) score (≥2 vs. 1 vs. 0), decreased HDL-C level (<40 mg/dL), elevated ß2-MG level, and response status after induction chemotherapy correlated significantly with poor progression-free survival (PFS) and overall survival (OS) (p < 0.05). In a multivariate Cox regression model that included IPI score, HDL-C level, ß2-MG level, and response status after induction chemotherapy, it was found that HDL-C level and response status after chemotherapy were independent prognostic factors for OS (p = 0.014 and 0.010, respectively) and PFS (p = 0.016 and 0.020, respectively). In conclusion, HDL-C was found to be a valuable independent prognostic factor in ENKTL, and the mechanism needs to be further investigated, which may offer the possibility of therapeutic targets.

[The protective effect of Xuebijing injection pretreatment on hepatic ischemia reperfusion injury and coagulopathy after excision of liver cancer].

OBJECTIVE: To observe the protective effect of Xuebijing injection pretreatment on hepatic ischemia reperfusion (I/R) injury and coagulopathy in liver cancer patients undergoing excision of hepatic cancer after occlusion of hepatic blood flow. METHODS: A prospective randomly controlled study was conducted. Sixty patients with liver cancer classified as Child-Pugh class A undergoing hepatectomy in the Department of Hepatobiliary Surgery of Sun Yat-sen University Cancer Center from October 2011 to March 2013 were enrolled. The patients were randomized into control group and Xuebijing group (each patient received 100 mL Xuebijing injection added to 0.9% saline as a preoperative treatment for 3 days). Complete blood count, coagulation function, hepatic function, serum pro-inflammatory cytokines and alpha-fetoprotein (AFP) levels were determined before and after operation. RESULTS: Forty-five out of 60 patients were enrolled eventually, with 23 patients in control group and 22 in Xuebijing group, and among them 43 patients were positive for hepatitis B surface antigen (HBsAg) at admission. Compared with those before operation, the postoperative levels of alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH) in control and Xuebijing groups were significantly elevated, prothrombin time (PT) and activated partial prothrombin time (AfYIT) were significantly prolonged, and white blood cells (WBC) , proportion of neutrophils (N) and C-reactive protein (CRP) were significantly increased (P<0.05 or P<0.01 ). Although the above indexes in Xuebijing group after operation were lower than those in control group in different degrees [ALT (U/L): 213.1 (80.4-796.6) vs. 265.8 (15.6-882.3), AST (UIL): 194.1 (65.4-914.2) vs. 264.3 (15.4-475.9), LDH (lg,U/L): 5.69 ± 0.72 vs. 5.71 ± 0.72, PT (s): 15.24 ± 2.16 vs. 14.41 ± 1.33, AfYIT (s): 31.51 ± 7.04 vs. 29.47 ± 4.90, WBC (x 109/L) : 13.4 7 ± 4.66 vs. 14.58 ± 4.40, N: 0.87 ± 0.06 vs. 0.87 ± 0.04, CRP (mg/L): 40.64 (16.93-189.59) vs. 45.64 ( 1.65-349.40) J, no statistical significance was found between the groups (all P>0.05 ). The preoperative levels of tumor necrosis factor-a (TNF -a) and interleukin-6 OL-6) were both less than 1.0 ng/L, and the postoperative levels of TNF-a showed no significant change, and IL-6 was increased to 485.10 (104.00-837.50) ng/L and 193.26 (95.10-385.20) ng/L in control and Xuebijing groups respectively (P<0.01). The serum high mobility group box-1 ( HMGB1 ) protein levels after operation were higher than those of preoperative in both groups (both P<0.01), but the postoperative HMGB1 in Xuebijing group were significantly lower than those in control group (j.Lg/L: 268.73 ± 5.56 vs. 277.12 ± 2.92, P<0.01). Acute physiology and chronic health evaluation ll (APACHE ll) score in Xuebijing group was significantly lower than that in control group (4.18 ± 3.75 vs. 4.53 ± 2.34, t=5.328, P=0.027), and the first passage of flatus and defecation after operation in Xuebijing group were significantly earlier than those in control group [exhaust time (days): 3 (2-4) vs. 3 (2-4), U=-2.023, P=0.043; defecation time (days): 4 (2-6) vs. 5 (3-8), U =-2.926, P=0.003 J. However, no difference was found between two groups in the postoperative and total hospital days. Spearman rank correlation analysis showed there were positive correlations between hepatitis B virus (HBV)-DNA levels and preoperative ALT (r=0.414, P=0.044) and AST (r=0.405, P=0.024) in 33 HBV-DNA positive patients, but there was no significant correlation between HBV -DNA levels or other preoperative liver function indicators. CONCLUSIONS: Hepatic I/R injury and coagulopathy may occur in liver cancer patients undergoing resection of cancer with occlusion of hepatic blood flow. Xuebijing injection may inhibit the release of serum pro-inflammatory cytokines, thereby alleviate hepatic I/R injury and promote the recovery of intestinal function. But it does not offer protective effect on coagulopathy.

Combined determination of Epstein-Barr virus-related antibodies and antigens for diagnosis of nasopharyngeal carcinoma.

BACKGROUND AND OBJECTIVE: With the development of molecular biology in recent years, many indexes for detecting Epstein-Barr virus (EBV) have been developed. This study was to evaluate the diagnostic value of combined determination of EBV-related antibodies and antigens, including VCA-IgA, EA-IgA, EBV-DNase antibody and EBV-DNA, in diagnosing nasopharyngeal carcinoma (NPC). METHODS: Serum and plasma samples from 160 untreated NPC patients and 76 healthy donors were collected. VCA-IgA and EA-IgA in the serum samples were detected by immunoenzyme staining method. Raji cells were stimulated by ortho-butanoic acid and croton oil to detect EBV-DNase antibody. The content of EBV-DNA in the plasma samples was detected by real-time fluorescence quantitative polymerase chain reaction (RQ-PCR). The diagnostic values of the indexes for NPC were evaluated. RESULTS: The sensitivity and specificity for diagnosing NPC were 90.0% and 89.5% for VCA-IgA, 75.0% and 94.7% for EA-IgA, 76.3% and 90.8% for EBV-DNase antibody, 68.8% and 88.2% for EBV-DNA, and 98.8% and 84.2% for combined determination. The positive rates of VCA-IgA and EA-IgA had no relationship with clinical stage of NPC (p > 0.05); nevertheless, the positive rates of EBV-DNase antibody and EBV-DNA were related with clinical stage (p < 0.05). CONCLUSIONS: The sensitivity of VCA-IgA and the specificity of EA-IgA are the highest while detecting solely. Combined determination could improve the diagnostic sensitivity and accuracy for NPC. EBV-DNase antibody and EBV-DNA could be helpful to evaluate the course of disease and classify the clinical stage of NPC.

[Interleukin-12 restores and promotes the T-cell immune function inhibited by 5-fluorouracil].

BACKGROUND & OBJECTIVE: Currently, 5-fluorouracil (5-FU) is still one of the widely applied chemotherapeutic agents for tumors. Interleukin-12 (IL-12) can promote the differentiation of Th1 cells and induce the production of interferon-gamma (IFN-gamma) by CD8+ T cells. This study was to investigate the suppression mechanism of 5-FU on the immune response mediated by T cells from normal human peripheral blood, and to determine the effect of IL-12 on the immune suppression induced by 5-FU. METHODS: The effects of 5-FU on the proliferation of peripheral blood mononuclear cells (PBMCs) and liver cancer cell line HepG2 were examined. PBMCs were stimulated with either anti-CD3 alone or anti-CD3 plus anti-CD28 in the presence or absence of 5-FU at different concentrations (0.20-50.00 microg/ml). The level of IFN-gamma in the culture supernatant was determined by ELISA. PBMCs were pretreated with 5-FU and stimulated with anti-CD3 and anti-CD28 for 2 days. The proportions of CD4+IFN-gamma+, CD8+IFN-gamma+, CD4+IL-2+ and CD8+IL-2+ T cells, and the expression of CD25 on CD4+ and CD8+ T cells were examined by flow cytometry (FCM). PBMCs were cultured in different combinations with anti-CD3 plus anti-CD28, IL-12 and/or 5-FU for 48 h. IFN-gamma level in the supernatant was detected by ELISA. The expression of IFN-gamma on CD4+ and CD8+ T cells were examined by FCM. RESULTS: 5-FU inhibited the proliferation of HepG2 cells and PBMCs, and suppressed INF-gamma production in PBMCs in a dose-dependent manner. The proportions of immune T cells were lower in 5-FU-pretreated PBMCs than in control PBMCs (0.7% vs. 2.1% for CD4+IFN-gamma+ T cells, 2.2% vs. 3.9% for CD8+IFN-gamma+ T cells, 0.7% vs. 2.5% for CD4+IL-2+ T cells, 0.2% vs. 0.4% for CD8+IL-2+ T cells). Both the positive rate and mean fluorescence intensity (MFI) of CD25 on CD4+ and CD8+ T cells were decreased after pretreatment of 5-FU. When stimulated by anti-CD3 and anti-CD28, the proportions of CD4+IFN-gamma+ and CD8+IFN-gamma+ T cells were 1.1% and 3.2% before adding IL-12, and 1.6% and 4.1% after treatment of IL-12. When stimulated by anti-CD3, anti-CD28, and 5-FU, the proportions of CD4+IFN-gamma+ and CD8+IFN-gamma+ T cells were 0.5% and 1.1% before adding IL-12, and 1.0% and 2.5% after treatment of IL-12. CONCLUSIONS: 5-FU could inhibit the proliferation of HepG2 cells and the immune function of PBMCs. IL-12 could restore the T-cell immune function inhibited by 5-FU.

[Detection of serum tumor supplied group of factors and its clinical significance].

BACKGROUND & OBJECTIVE: Tumor supplied group of factors (TSGF) is the first kind of tumor marker in vitro which was primarily obtained the approval by Chinese government to land the market, there was no any reports about it in abroad before. In order to know its specificity and sensitivity, we designed this study to detect the TSGF in serum of the patients with malignancies and evaluate its clinical significance for diagnosis. METHODS: Serum TSGF in 523 cases with malignancies and 400 cases of normal control were detected with chemical colorimetry. The diagnostic accuracy was evaluated. RESULTS: Mean levels of serum TSGF in the patients without malignancies (group A), the patients with malignancies (group B), the patients after curative treatment for malignancies (group C) and the normal control (group D) were 66.57, 71.24, 60.78, and 53.56 u/ml, respectively. After the statistical analysis, differences between the group A and group B, group A and group D, group B and group C, group B and group D, group C and group D were found (P < 0.05). But there was no obvious difference between the group A and group C (P > 0.05). The sensitivity was 63.86%; the specificity was 90.89%; the truly negative rate was 87.01%; and the falsely negative rate was 13.00%. CONCLUSIONS: Serum TSGF increased in malignancies and may be helpful for screening and early detection of cancer.