Prognostic significance of serum anti-Epstein-Barr virus antibodies in nasopharyngeal carcinoma / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the association between serum against Epstein-Barr virus (EBV) antibodies levels and nasopharyngeal carcinoma (NPC) patients' prognosis.</p><p><b>METHODS</b>Blood samples from 140 primary NPC patients without metastasis were collected before and after treatment. The titers of VCA/IgA and EA/IgA were detected by immunoenzyme assay, and the levels of NA1/IgA and Rta/IgG were detected by enzyme-linked immunosorbent assay (ELISA). All patients received consequent follow-up and long-term efficacy and survival assessment.</p><p><b>RESULTS</b>Post-treatment serum levels of VCA/IgA, EA/IgA, NA1/IgA and Rta/IgG in NPC patients significantly decreased than those before treatment, while had significantly higher than those in control individuals (P < 0.05). Patients in remission had significantly lower pre-treatment serum levels of VCA/IgA and EA/IgA than patients with progression (P < 0.05). None of serum levels of VCA/IgA, EA/IgA, NA1/IgA and Rta/IgG was associated with the 3-year overall survival (P > 0.05). The progression-free survivals were significantly lower in patients with higher pre-treatment VCA/IgA (> or = 1 : 320) and EA/IgA (> or = 1:80) levels than in those with lower VCA/IgA ( < 1 : 320) and EA/IgA (< 1 : 80) levels, respectively (61.8% vs. 86.5% , 61.3% vs. 86.5%, P < 0.001). Cox regression model analysis demonstrated that pre-treatment serum VCA/IgA level was an independent risk factor for progression-free survival (HR = 3.80, P = 0.001).</p><p><b>CONCLUSION</b>Anti-EBV VCA/IgA and EA/IgA might provide information regarding the prognosis of NPC patients.</p>

Evaluation of computational methods for HLA three loci haplotype by compare with family-based data / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>We evaluated the accuracy and efficiency of computational inference methods for haplotype on estimate HLA-A-B-C haplotype frequencies by compared with the haplotypes manually defined in a family-base dataset.</p><p><b>METHODS</b>558 individuals with pedigree information were selected, and their haplotyps were compared with the data obtained by the following three method: the Expectation-Maximization (EM) and Excoffier-Laval-Balding (ELB)algorithms using the AELEQUIN software, and the SAS/Genetics PROC HAPLOTYPE method.</p><p><b>RESULTS</b>After performing the SAS/Genetics method, and the Expectation-Maximization (EM) and Excoffier-Laval-Balding (ELB) algorithms using the AELEQUIN software, 248, 247, and 238 different haplotypes were obtained respectively. The accuracy rates of these three methods were 88.5%, 89.1%, and 90.3% respectively. There are no significant different in the accuracy and estimated haplotype frequency comparisons among any two of these computational inference methods.</p><p><b>CONCLUSION</b>High accuracy haplotype frequency estimate rates could be obtained by these three computational inference methods, and there are no significant difference in the comparison of haplotypes estimated by SAS/Genetics, the EM and ELB algorithms using the AELEQUIN software. However, ELB algorithm shows better performance than EM algorithm and SAS/Genetics PROC HAPLOTYPE method for haplotype frequencies estimation in general.</p>

Relationship between clinical stages of nasopharyngeal carcinoma and Epstein-Barr virus antibodies Rta/IgG, EBNA1/IgA, VCA/IgA and EA/IgA / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the relationship between the clinical stages of nasopharyngeal carcinoma (NPC) and Epstein-Barr virus (EBV) antibodies Rta/IgG, EBNA1/IgA, VCA/IgA and EA/IgA.</p><p><b>METHODS</b>Serum samples obtained from 211 untreated patients with NPC categorized by the project of 92' stage were examined for the presence of the EBV antibodies Rta/IgG and EBNA1/IgA by enzyme-linked immnunosorbent assay (ELISA) and for VCA/IgA and EA/IgA by immunoenzymatic assay. The positive rates and antibody levels in the NPC patients in different TNM stages and clinical stages were analyzed statistically.</p><p><b>RESULTS</b>No significant difference in Rta/IgG rA value was found in the NPC patients in different TNM or clinical stages (P>0.05). The EBNA1/IgA rA value was significantly lower in stage T1, N0, and clinical stage I than in the other corresponding T stages, N stages and other clinical stage (P<0.05). The antibody titers of VCA/IgA and EA/IgA differed significantly between the N stages and the clinical stages (P<0.05).</p><p><b>CONCLUSION</b>The expression of EBV Rta/IgG is not associated with NPC stage. The expression of EBNA1/IgA is relatively low in early NPC. The antibody level of VCA/IgA and EA/IgA are significantly correlated to the degree of neck lymph node metastasis, and might be helpful to classify the clinical stages of NPC.</p>

Combined detection of Epstein-Barr virus antibodies for serodiagnosis of nasopharyngeal carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of combined detection of Epstein-Barr virus (EBV) VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA in serodiagnosis of nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Serum samples obtained from 211 untreated patients with NPC and 203 non-NPC ENT patients were examined for the presence of VCA/IgA and EA/IgA by immunoenzymatic assay and for Rta/IgG and EBNA1/IgA by enzyme-linked immunosorbent assay (ELISA). The receiver operating characteristic (ROC) curve was generated to confirm the cutoff values of different antibodies. The evaluation indexes of combined detection of multiple antibodies used for serodiagnosis of NPC were calculated with compounded positive judgment method.</p><p><b>RESULTS</b>Compared to a single antibody, combined detection achieved a higher sensitivity and specificity. The sensitivity of VCA/IgA + Rta/IgG + EBNA1/IgA (98.1%) was higher than the other 3 combinations with a specificity, accuracy, Youden index and positive predictive value (PPV) of 88.7%, 93.5%, 0.868 and 90.0%, respectively. The combination of EA/IgA+Rta/IgG+EBNA1/IgA had the highest specificity (95.1%), accuracy (94.9%), Youden index (0.899) and PPV (95.2%), with a sensitivity of 94.8%, suggesting its higher accuracy in the serodiagnosis of NPC. Combined detection of the 4 antibodies had the highest sensitivity (98.6%) with a specificity, accuracy, Youden index and PPV of 88.2%, 93.5%, 0.868 and 89.7%, respectively.</p><p><b>CONCLUSIONS</b>Combined detection of Rta/IgG against immediate early antigens, EA/IgA against early antigens, VCA/IgA against late antigens, and EBNA1/IgA against latent antigens provides better understanding of the expression profiles of EBV lytic and latent antigens with excellent complementarity, and may serve as an optimal combination for NPC serodiagnosis.</p>

The experimental study on anti-tumor effect of 131Ⅰ-Tyr-octreotide in nude mice bearing human non-small cell lung cancer / 中华核医学杂志

Objective Radionuclide-labeled low molecular weight polypeptide is reeently advocated for the diagnosis and treatment of malignant tumor. The purpose of this study was to evaluate the anti-tumor effect of 131Ⅰ-Tyr-octreotide in nude mice bearing human non-small cell lung cancer (NSCLC). Methods 131Ⅰ-Tyr-octreotide was prepared by Ch-T method. The radiochemical purity was measured and biodistribution was evaluated. The nude mice models bearing human NSCLC were studied and divided into four groups: group A injected 131Ⅰ-Tyr-octreotide through tail vein, group B injected normal saline, group C injected 131Ⅰ-Tyroctreotide through stroma and group D injected 131Ⅰ through stroma. The radioactivity ratio of tumor to normal tissue (T/NT) was calculated over region of interest (ROI). The tumor cell cycle and cell apoptosis were analyzed by flow cytometry (FCM), terminal deoxynucleotidyl transferase mediated dUTP-biotion nick end labeling (TUNEL) and histopathological analysis. Statistical analysis was performed with SPSS 11.0, and the comparison for difference between groups performed with one-way ANOVA analysis. Results The labeled radiochemical purity was (95.23±1.67)% and specific activity of 3.5×106Bq/ug. The biodistributiou showed high uptake in kidney, and low uptake in liver and spleen. The radioactive uptake in group C was higher than the other groups, and the retention time was longer. The T/NT was 52.74±0.13 after 24 h, which was much higher than that the other groups (group D: 8.90±0.23, group A: 6.42±0.02, q=628.81 and 664.33, all P<0.05). The resuits of tmnor cell cycle determined by FCM showed that the G1 phase was blocked mast remarkably in group C than the other groups [group C: (83.17±6.86)%, group A: (57.02±18.81)%, group D: (49.29±7.80)%, group B: (45.88±5.13)%, q=5.29, 6.86, 7.55, 1.56, 2.26, 0.69, all P<0.05]. Apeptotic cells were observed by TUNEL, and apoptotic body was detected by immuno-histochemical examination. Conclusions 131Ⅰ-Tyr-octreotide was easily labeled by Ch-T. 131Ⅰ-Tyr-octreotide could induce tumor cell apoptosis and inhibit the tumor cell of NSCLC. It might be a potential target-directed agent in NSCLC.

Evaluation of detection of Epstein-Barr virus Rta/IgG in nasopharyngeal carcinoma / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the clinical value of Epstein-Barr virus (EBV) Rta/IgG in the diagnosis of nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Serum samples derived from 211 untreated patients with NPC, 413 subjects including 203 non-NPC ENT patients and 210 healthy volunteers as control were examined for the presence of antibodies directed against Rta/IgG by using enzyme-linked immnunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve was applied to perform methodical evaluation of this tumor marker.</p><p><b>RESULTS</b>The rA value median of Rta/IgG in NPC group was significantly higher than one in control group (P < 0.001). The area under ROC was 0.933. The sensitivity and specificity of this marker were 90.5% and 90.1%, respectively, when the best cutoff value was defined.</p><p><b>CONCLUSION</b>Rta/IgG detected with ELISA method is a new target of EBV, and may be one of important marker for NPC diagnosis.</p>

Evaluation of combined determinations of Epstein-Barr virus antibodies for nasopharyngeal carcinoma assessed with receiver operating characteristic curve based on logistic regression / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the diagnostic value of combined determination of Epstein-Barr virus (EBV) antibodies for nasopharyngeal carcinoma (NPC), including immunoglobulin (Ig) A against EBV capsid antigens (VCA), IgA against early antigens (EA), IgG against BRLF1 transcription activator (Rta) and IgA against EBV nuclear antigen-1 (EBNA1), assessed with receiver operating characteristic (ROC) curve based on logistic regression.</p><p><b>METHODS</b>Serum samples derived from 211 untreated patients with NPC and 203 non-NPC ENT patients were examined for the presence of VCA/IgA and EA/IgA by immunoenzymatic assay, Rta/IgG and EBNA1/IgA by enzyme-linked immnunosorbent assay (ELISA). The different Logistic regression models were established for various combined determinations of antibodies, respectively. Using the predicted probability as the analyzed variable, ROC curve was applied to evaluate the diagnostic accuracy of different combined determinations.</p><p><b>RESULTS</b>The sensitivity of VCA/IgA (98.1%) and the specificity of EA/IgA (98.5%) were the highest while detecting solely. The results which were analyzed by ROC curve based on Logistic regression showed that the sensitivity and specificity were improved. In two-marker combinations, VCA/IgA + Rta/IgG whose area under ROC curve (AUC) was 0.991 had the highest diagnostic accuracy, and its sensitivity, specificity and Youden index were 94.8%, 98.0% and 0.928 respectively. No significant difference of AUC were found comparing VCA/IgA + Rta/IgG with VCA/IgA + Rta/IgG + EBNA1/IgA and four-marker combination( P > 0.05), of which sensitivity, specificity and Youden index were 94.8%, 98.5%, 0.933 and 96.7%, 97.0%, 0.937, respectively.</p><p><b>CONCLUSION</b>The approach of ROC curve based on Logistic regression can improve synthetic efficiency for combined determination of multiple markers. The combined determination of VCA/IgA and Rta/IgG with a complementary effect is optimal for NPC serodiagnosis.</p>

Molecular detection and haematological analysis of heterozygotes in beta-thalassemia combining deletional alpha-thalassemia / 中国实验血液学杂志

This study was aimed to investigate the prevalence and genotype distribution of heterozygotes in beta-thalassemia combining deletional alpha-thalassemia by using molecular detection and haematological methods. Three common deletions of alpha-thalassemia were detected by using gap-PCR. The mutations of beta-thalassemia were identified by using PCR with reverse dot blot hybridization. The routine analysis of blood cells was carried out. The results indicated that 15 cases from the 81 beta-thalassemia traits were found to be the compound heterozygosity for beta-thalassemia and alpha-thalassemia with 9 different types of gene defects with 18.52% detection rate. There were 6 cases (7.41%) of beta-thalassemia heterozygote combining alpha-thalassemia-1 gene (--(SEA)/alphaalpha), 8 cases (9.88%) combining with alpha-thalassemia-2 gene including 6 (7.41%) right ward deletion (-alpha(3.7)/alphaalpha) and 2 (2.47%) left ward deletion (-alpha(4.2)/alphaalpha), and 1 case (1.23%) combining deletional HbH gene (--(SEA)/-alpha(3.7)). No significant differences were found between beta-thalassemia heterozygotes combining deletional alpha-thalassemia and pure beta-thalassemia in all RBC parameters. It is concluded that the incidence of beta-thalassemia heterozygotes combining with deletional alpha-thalassemia is frequent in Wuzhou city. The hematological analysis can not give specificity for diagnosing these dual heterozygotes. Gap-PCR as a routine method for thalassemia screening has the advantages in reducing the possibility of failing to detect the combining heterozygosity for beta-thalassemia and alpha-thalassemia. It is more useful for genetic counselling and prenatal diagnosis of this disease.

Sequence analysis of the deletion and mutation in carboxy terminal region of the Epstein-Barr virus latent membrane protein 1 derived from nasopharyngeal carcinoma patients / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the deletion and mutation in carboxy terminal region of LMP1 gene derived from nasopharyngeal carcinoma (NPC) in Guangdong and Guangxi, the high risk areas of nasopharyngeal carcinoma in China.</p><p><b>METHODS</b>LMP1 gene carboxy terminal region was amplified from nasopharyngeal carcinoma tissues by PCR, and then cloned and sequenced.</p><p><b>RESULTS</b>Of the 20 cases, 17 were LMP1 positive. In all positive cases, only 1 case did not show deletion. Four positive cases were chosen for DNA sequencing, The rusult showed that all the four cases had mutation and the 30bp deletion.</p><p><b>CONCLUSIONS</b>High frequency of deletion and mutation in LMP1 gene of nasopharyngeal carcinoma tissues was found in Guangdong and Guangxi. Whether it related to the high incidence of NPC should be further studied.</p>