Assessment of EBV antibodies and EBV-DNA in the diagnosis and stages of nasopha-ryngeal carcinoma / 中国肿瘤临床

Objective:To evaluate the efficacy of Epstein-Barr nuclear antigen 1/immunoglobulin A (EBNA1/IgA), BamH1 Z transactivator/IgA (Zta/IgA), capsid antigen/IgA (VCA/IgA), and Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) in detecting different stages of na-sopharyngeal carcinoma (NPC). The relationship between the EBV markers and stages of NPC was also analyzed. Methods:Blood sam-ples of 152 untreated patients with NPC and 675 healthy subjects were collected.ELISA was used to detect the serum levels of EBNA1/IgA, Zta/IgA, and VCA/IgA. Fluorescence quantitative PCR (FQ-PCR) was used to detect the plasma levels of EBV-DNA. ROC and correla-tion analyses were employed to assess the detection assays for NPC diagnosis. The positive rates of EBV markers in NPC patients in dif-ferent stages were analyzed statistically. Results: The positive rates of EBNA1/IgA, Zta/IgA, VCA/IgA, and EBV-DNA in NPC patients were higher than those in the healthy individuals. The expression of EBNA1/IgA was relatively high in early NPC. The sensitivity of EB-NA1/IgA was 77.8%. In advanced NPC, the level of EBV-DNA was high, and the sensitivity of EBV-DNA was 88.8%. The specificity of EBV-DNA and EBNA1/IgA could reach more than 96%. The combination of EBV-DNA and EBNA1/IgA showed the best diagnostic value, with a sensitivity of 92.1%(early stage 82.5%, advanced stage 98.9%) and a specificity of 96.9%. The positive rates of EBV-DNA were positively associated with the NPC clinic stage and N stage. The positives rates of Zta/IgA were positively associated with the NPC N stage. Conclusion:The best single index for NPC screening in an asymptomatic population is EBNA1/IgA. EBV–DNA is an ideal index for auxiliary diagnostics of advanced NPC. The combination of EBV-DNA and EBNA1/IgA shows the best diagnostic value. EBV-DNA is an important index in the stage and illness monitoring of NPC. Zta/IgA can indirectly reflect the character of lymph node metastasis, and it may be useful in assessment of NPC surveillance.

Embryonic stem cell test for the study of the embryotoxicity of hydroquinone / 国际检验医学杂志

Objective To establish a model of embryonic stem cell test(EST)and utilize this model to evaluate the embryotoxici-ty of hydroquinone.Methods Mouse 3T3 fibroblasts and mouse embryonic stem(ES)cells(ES-E14TG2a)were cultured in vitro, and methyl thiazolyl tetrazolium(MTT)test was performed to detect the cytotoxicity of 3T3 cells and ES-E14TG2a cells induced by the positive control(5-fluorouracil),negative control(penicillin G)and tested compound(hydroquinone).The concentrations of the test compounds that inhibited 50% viability of ES-E14TG2a cells(IC50 ES)and 3T3 fibroblasts (IC50 3T3)were calculated.The hanging-suspension-adherent culture systems were used to induce embryonic stem cells into cardiomyocytes,and the concentrations of test compounds that caused 50% inhibition of differentiation of ES-E14TG2a cells into cardiomyocytes (ID50 ES)was calculated. The embryotoxic potential of hydroquinone was classified by prediction model of the embryonic stem cell test.Results The prolif-eration of ES-E14TG2a and 3T3 cells were inhibited by hydroquinone,of which the IC50 3T3 and IC50 ES values were (5.97±0.48) and (2.57±0.10)μg/mL respectively.The differentiation of ES-E14TG2a cells were also inhibited by hydroquinone,of which the ID50 ES was (3.77±0.31)μg/mL.Hydroquinone was evaluated as a strong embryotoxicity chemical by prediction model of EST. Conclusion Hydroquinone exhibits a strong embryotoxicity.

Intraarticular injection of autologous bone marrow mesenchymal stem cells for mild-to-moderate osteoarthritis / 中国组织工程研究

BACKGROUND:Increasing evidence col ected from animal experiments or laboratories shows that bone mesenchymal stem cel s possess potent immunosuppression and anti-inflammation effects and cartilage regenerative capability. The microenvironment in human knee joint of osteoporosis is more complex and involves lots of bioactive factors and immunologic mechanisms. OBJECTIVE:The analyze the therapeutic effects of intra-articular injection of autologous bone marrow mesenchymal stem cel s on mild-to-moderate osteoporosis, and to investigate the mechanisms for anti-inflammation, immunoregulation and reversion of cartilage degradation. METHODS:About 15 mL bone marrow was aspired from 26 patients with mild-to-moderate osteoporosis and taken to the laboratory where bone marrow mesenchymal stem cel s were isolated and characterized in terms of some surface markers by a flow cytometer, and the ability of osteogenic and adipogenic differentiation was analyzed. The bilateral knees of each patient were divided into two groups at random. The experiment group were treated with autologous bone marrow mesenchymal stem cel s by intra-articular injection (2×107 cel s), while the control group injected the same volume of control medium without bone marrow mesenchymal stem cel s. Western Ontario and McMaster Universities Osteoarthritis Index was used to estimate the function of the knee joints before and after the treatment. We col ected the joint fluid before, 2 and 4 weeks post treatment, and then measured the production of interleukin-1, interleukin-10, tumor necrosis factor-α, cartilage oligomeric matrix protein using ELISA. RESULTS AND CONCLUSION:According to the standardized culture-expansion protocol, a sufficient number of bone marrow mesenchymal stem cel s (more than 2×107 passage 3 cel s) were obtained for intra-articular injection. The bone marrow mesenchymal stem cel s isolated from patients were positive for CD105, CD29 and negative for CD45, CD34, and had the fair osteogenic and adipogenic capacities. The function of the knee joints was improved obviously after treatment. In the experimental group, the secretion of both interleukin-1, tumor necrosis factor-αand cartilage oligomeric matrix protein in the joint fluid was suppressed, while the interleukin-10 level was up-regulated compared to the control group. The results suggest that intra-articular injection of autologous bone marrow mesenchymal stem cel s can exert good effect in mild-to-moderate osteoporosis patients. Furthermore, bone marrow mesenchymal stem cel s via intra-articular injection can function through anti-inflammation and reversion of degradation of the articular cartilage, which is a new promising approach for treating mild-to-moderate osteoarthritis.

Embryotoxicity of eugenol based on a model of embryonic stem cell test / 中国组织工程研究

BACKGROUND:As the pharmacological effect of eugenol constantly being discovered, its application in medical and food industry becomes wider. However, its toxicity studies have not established a complete database, especialy in the improvement of safety assessment of developmental toxicity and teratogenicity. OBJECTIVE:To establish a model of embryonic stem cel test to evaluate the embryotoxicity of eugenol. METHODS:Mouse fibroblasts (3T3) and mouse embryonic stem cels (E14TG2a) were culturedin vitro, and MTT test was performed to detect the cytotoxicity of 3T3 cels and E14TG2a cels with positive control 5-fluorouracil, negative control penicilin G and tested compound eugenol. The concentration of the tested compounds that inhibiting 50% viability of embryonic stem cels (IC50 E14TG2a) and 3T3 fibroblasts (IC50 3T3) was calculated. The hanging-suspension-adherent culture systems were used to induce embryonic stem cels into cardiomyocytes, and the concentration of tested compounds that caused 50% inhibition of differentiation of E14TG2a cels into cardiomyocytes (ID50 E14TG2a) was calculated. The embryotoxic potential of eugenol was classified by prediction model of the embryonic stem cel test. RESULTS AND CONCLUSION:The proliferations of E14TG2a and 3T3 cels were inhibited by eugenol, of which the IC50 3T3 and IC50 E14TG2a values were (3.613±0.192) and (1.799±0.131) mg/L. The differentiation of E14TG2a was also inhibited by eugenol, of which the ID50 E14TG2a was (3.501±0.158) mg/L. Eugenol was evaluated as a chemical compound with strong embryotoxicity by the model of embryonic stem cel test.

Early NPC detection through serologic risk stratification and clinical follow-up of high risk subjects / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Nasopharyngeal Carcinoma (NPC) can be successfully treated by radiotherapy, if the tumor is confined to nasopharynx, but clinical onset is usually delayed to more advanced stages, when prognosis is poor. The objective is to determine efficacy of a new program for early NPC detection, which entails stratification of the NPC risk of target population according to serum levels of 3 Epstein Barr Virus (EBV) antibodies.@*METHOD@#The sera of 1373 healthy adult residents from Zhongshan were collected and analyzed in this study from Mar 16, 2007 to Dec 31, 2007. The levels of EBNA1/IgA, zta/IgG and EBNA1/IgG were tested by ELISA. To stratify the subjects of 1373 adults into high, moderate and normal NPC risk groups by regression analysis of the levels of the EBV antibody. The high-risk groups of nasopharyngeal carcinoma risk could be followed-up every 3-6 month.@*RESULT@#NPC risk of 1379 adults was stratified according to serum levels of the 3 EBV antibodies. Eleven (0.8%) were identified to be of high risk for NPC, having high levels of all three antibodies and/or IgA EBNA level > 3 rod. Clinical examination of high risk subjects detected 5 NPC cases, 3 cases detected in the first instance and 2 in follow-up examination 3 to 6 months hence. Three cases were diagnosed with UICC Stage I tumor (60%), one in the first instance and 2 in follow-up, and the 5 cases account for all NPC cases detected from the entire cohort over 28 months(100%).@*CONCLUSION@#The new program affords an efficient and efficacious means for early NPC detection.

Assessment of nasopharyngeal carcinoma risk by EB virus antibody profile / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the risk of nasopharyngeal carcinoma (NPC) through EB virus antibody profile by enzyme linked immunosorbent assay (ELISA).</p><p><b>METHODS</b>EBNA 1/IgA, EBNA 1/IgG and zta/IgG by ELISA and VCA/IgA by immmunoenzymatic method were detected in 121 NPC patients and 332 healthy subjects (HS) in the Pearl river estuary.</p><p><b>RESULTS</b>The sensitivity rates were 85%, 83% and 79% for EBNA 1/IgA, EBNA 1/IgG and zta/IgG, all three of which if combined was the highest 92%. The specificity rates were 86%, 86% and 80% for EBNA 1/IgA, EBNA 1/IgG and zta/IgG, all three of which if combined was also the highest 93%. According to the level of odds ratio, nasopharyngeal carcinoma risk could be divided into 3 groups: low, moderate and high-risk groups. 93% of HS had low risk of NPC with the odds ratio 0.0 to 0.3. 0.4% of HS had high risk of NPC with the odds ratio of 137.9%.</p><p><b>CONCLUSION</b>ELISA is more objective than the traditional immunoenzymatic method in the detection and diagnosis of NPC. The combination of EBNA 1/IgA, EBNA 1/IgG and zta/IgG is able to evaluate the risk of NPC.</p>

Screening out nasopharyngeal carcinoma by two-stage ELISA for EB virus / 中国免疫学杂志

Objective:Screening out NPC from susceptable crowd by serological test for EB virus.Methods:Checking antibody of EB virus by ELISA. At first, EBNA1 IgA was detected for susceptable crowd as a primary screening index, secondary , EBNA1 IgG and Zta IgG were detected in the EBNA1 IgA positive group.Results:The sensitivity and specificity of EBNA1 IgA were 91.8% and 91.4%, both were higher than those of EBNA1 IgG and Zta IgG; the sencondary detection of EBNA1 IgG and Zta IgG raised the specificity of NPC screening to 96.5% and also helped to divide the crowd into 3 groups as high, median, and low risk. And the high risk group accounted for 0.39%.Conclusion:Two-stage screening of NPC raise the sensitivity and specificity of NPC detection. It also helps to divide the crowd into 3 groups of risk.

The changes of IL-6 gene expression in acute repetitive hypoxia / 中华麻醉学杂志

Objective: This study was to observe the changes of IL-6 gene expression in acute repetitive hypoxia. Method: When the ratio of living cells was more than 95%, nerve cells cultured were passed through with mixed gases of 95% N_2+5%CO_2 for 3 min followed by another gases mixture of 95% O2+5% CO_2 for 10 min. After repeated the experiment as above, the alteration in the expression of IL-6 gene was measured using the RT-PCR method. The prod ucts of PCR were analyzed with computer Gel imaging and image analysis instrument. Result: After the first hypoxia IL-6 gene expression enhanced, and after the second hypoxia, it was still stronger than baseline although it declined slightly. There was no significant RNA disintegration. Conclusion:The IL-6 involves in the cellular defense in hypoxia adaptation.