The early diagnostic value of EBV-DNA load in plasma and PBMC for children′s primary infectious mononucleosis / 中华检验医学杂志

Objective To investigate the early diagnostic value of EBV-DNA load in Plasma and peripheral blood mononuclear cells (PBMCs) for Children′s Primary Infectious Mononucleosis(IM).Methods The plasma and PBMCs samples were collected from children of 67 cases with primary IM (IM groups) and 75 healthy cases (H groups) during April, 2015 to March, 2016 in our hospital. The EBV-DNA load in PBMCs and plasma samples were quantified by Real-time PCR. Statistical differences between two groups were analyzed by using Mann-Whitney Test. Receiver operating characteristic curves (ROC) were analyzed to assess the early diagnostic value of EBV-DNA load for primary IM.Results Levels of EBV-DNA in the samples of plasma were significantly higher in the IM groups (median, 380 IU/ml) compared to the H groups (Z=-9.332,P<0.01), and levels of EBV-DNA in the PBMCs samples were also significantly higher (median, 2.51×105 IU/ml, Z=-9.194,P<0.01). According to ROC curve analysis, the AUC of EBV-DNA load in plasma was 0.908, the most appropriate cut-off value was 12.5 IU/ml (sensitivity: 83.6% and specificity: 94.7%),the AUC of EBV-DNA load in PBMCs was 0.944, the most appropriate cut-off value was 2.19×104 IU/ml (sensitivity: 89.6% and specificity: 82.7%), the AUC of combined testing(plasma and PBMCs) was 0.967(sensitivity: 98.3% and specificity: 99.0%).Conclusions Combined testing of plasma and PBMCs have better clinical value, it may be useful for the diagnosis of early stage of Children′s Primary Infectious Mononucleosis.

Value of peripheral NLR and PLR for the survival of patients with primary gastric cancer / 中国生化药物杂志

Objective To investigate value of peripheral NLR and PLR for the survival of patients with primary gastric cancer.Methods The clinical data of 132 primary gastric cancer patients and 30 healthy controls were analyzed by the Kaplan-Meier, Log-rank test and multivariate COX regression.Results NLR, PLR levels of the case group were significantly higher than that in the healthy control group (t=6.67, P=0.000;t=13.23, P=0.000); the higher the age, the greater tumor diameter, the higher the degree of differentiation, lymph node metastasis, and not be treated with surgery, NLR and PLR could increase (P<0.05);NLR and PLR showed a significant positive correlation (r=0.3164, P=0.0002);survival time of low NLR group was (57.59 ±2.23) months and high NLR group was (35.22 ±3.09) months(P<0.05);survival time of low PLR group was (54.09 ±2.66) months and high PLR group was (35.22 ±2.75 ) months(P<0.05);age, clinical stage, lymph node metastasis and NLR, PLR levels were independent factors for the overall survival in patients with gastric cancer ( P<0.05 ) .Conclusion NLR and PLR of gastric cancer patients increase significantly and are closely related to tumor size, metastasis, clinical stage, and the deterioration, which showes some predictive value for the survival prognosis of the patients.

Construction and Expression of DNA Vaccine pIRES-Sj97-Sj14-Sj26 and Its Immunogenicity in Mice / 华中科技大学学报（医学）（英德文版）

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was trans-fected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA,plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh.Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the plRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ, level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P<0.01,P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice.

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes / 华中科技大学学报（医学）（英德文版）

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Construction and expression of bivalent membrane-anchored DNA vaccine encoding Sjl4FABP and Sj26GST genes / 华中科技大学学报（医学）（英德文版）

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.