Objective:To investigate the quantitative expression of Melanoma-associated antigen MAGE-E1 mRNA in glioma, and explore its potential for immunotherapy in glioma.Methods:To establish a quantitative real-time polymerase chain reaction ( qRT-PCR) method to quantitatively determine MAGE-E1 mRNA in glioma.A total of 47 human glioma and 14 normal brain tissue specimens were analyzed.MAGE-E1 mRNA was normalized to HPRT1, a housekeeping gene.MAGE-E1/HPRT1 was used to evaluate the expression level of MAGE-E1 mRNA.Results:High-level expression of MAGE-E1 was observed in 7.1%of normal brain and 66.0%glioma tissues,which shown significant difference with P<0.05.There was irrelevant between the expression of MAGE-E1 mRNA and clinicopathogical parameters ,such as gender ,age,histological subtype and grade.Conclusion:With higher expression of MAGE-E1 in glioma tissues,it might be a potential target for immunotherapy of glioma.

The morphogenesis of Valsalva's sinus of semilunar valves in chick embryo was studied with SEM and LM. The sinus began to develop at stage 30 and was basically formed at stage 34 (based on H-H staging). The endothelial cells on arterial and ventricular surfaces of the valve were morphologically different. Mesenchymal cells beneath the endothelium were distributed loosely or densely on different parts of the valve in accordance with the morphological changes of the endothelial cells. The study suggests that the formation of the sinus is associated with not only the growth of endothelial cells on the arterial surface of the valve into underneath mesenchyme, but also with the mesenchyme which is loosing and presents some cavities.

Objective To analyze the percentage and functional changes of natural killer T (NKT) cells in peripheral blood and bone marrow of severe aplastic anemia (SAA) children before immunosuppressive therapy (IST) comparing to that of healthy children.Methods Ten children with severe aplastic anemia were included in the study and ten healthy children at the same age were selected as the control group. By lfow cytometry, the percentage of CD3+CD1d tetramer+ NKT cell in peripheral blood and bone marrow were detected from March 2014 to December 2014 in our hospital. Immune magnetic bead separation was used to isolate and purify iNKT cells .The puriifed iNKT cells were cultured in the OCH(50 ng/ml,100 ng/ml or 200 ng/ml)+rhIL-2+rhG-CSF culture systems. The ampliifcation of iNKT cells after cultured in different systems were calculated. Elispot method was used to analyze the spotting form cells (SFCs) of IFN-γ or IL-4 expressed by activated iNKT cells.Results The percentage of CD3+CD1d tetramer+ NKT cells in peripheral blood of SAA group(0.72±0.03)% was signiifcantly lower than that of the control group(0.92±0.02)%(P=0.000). The percentage of CD3+CD1d tetramer+ NKT cells in bone marrow of SAA group(0.82±0.02)% was signiifcantly lower than that of the control group(1.05±0.05)%(P=0.000).In vitro iNKT cell ampliif-cation ability of bone marrow in SAA group was signiifcantly lower than the control group, and in medium concentration(50±6) and high concentration OCH group(52±6), the ampliifcation ability was higher than that in low concentration OCH group(30±5) (P<0.05). The secretion of IFN-γ in the iNKT cells of SAA bone marrow was signiifcantly lower in medium concentration(33±3) and high concentration(35±3)OCH group than that of the low concentration(50±3)OCH group(P<0.01). The secretion of IL-4 in the iNKT cells of SAA bone marrow was signiifcantly higher in medium concentration(50±3)and high concentration(75±3) OCH group than that of the low concentration(33±3) OCH group(P<0.01).Conclusions The quantity and function of NKT cells from children with SAA are lower than that of the healthy children.In vitro, they had better ampliifcation ability and could improve IL-4/IFN-γ imbalance in medium concentration and high concentration OCH group than in low concentration OCH group.

10.3969/j.issn.1000-3606.2013.06.007

F-asparaginask(F-LSP)is onk of thk cke agknts in thk long-tkrm chkmothkrape of lemphoid malignanciks in childrkn with acutk lemphoblastic lkuckmia(LFF)and non Hodgcin＇s lemphoma(NHF). If F-LSP rksistanck occurs in patiknts,it is highle suggkstivk of poor prognosis. Thkrkfork,thk mkchanism of F-LSP rksistanck is a major stude in thk fikld of diagnosis and trkatmknt of lkuckmia in childrkn. Ovkr thk ekars,rklatkd studiks havk shown that thk bask lkvkl of asparagink senthktask in tumor cklls,bonk marrow hkmatopoiktic support cklls,somk advkrsk mkta-bolic changks aftkr F-LSP chkmothkrape,and thk F-LSP silkncing inactivation induckd be sklf nkutralizing antibode can lkad to F-LSP rksistanck. In this papkr,according to thk litkraturk rklatkd rkports in rkcknt ekars,providk rkfkrknck for domkstic collkaguks,in ordkr to carre out thk rklkvant basic and clinical rkskarch,to providk thkorktical basis for thk rkalization of individualization of chkmothkrape.

OBJECTIVE: To investigate the gene expression of Cancer-Testis Antigen SCP-1 in human nasopharyngeal carcinoma. METHOD: The SCP-1 mRNA were examined in the tumor tissues of 28 nasopharyngeal carcinoma cases and control pharynx vasitis tissues of 40 non-cancer cases by reverse transcriptase polymerase chain reaction. A randomized sample of positive expression of SCP-1 mRNA was extracted for DNA sequencing to examine the reliability of results. The correlation of the expression of SCP-1 mRNA with clinical data was analyzed by statistical method. RESULT: The expression frequency of SCP-1 mRNA was 14.3% (4/28) in nasopharyngeal carcinoma and zero (0/40) in control pharynx vasitis tissue of non-cancer cases (P < 0.05); The DNA sequencing confirmed that the product of reverse transcriptase polymerase chain reaction was truly the target gene; No significant relationship was found between the expression of SCP-1 mRNA and clinical characters, such as age, gender and serum anti-EBV level (P > 0.05). CONCLUSION SCP-1 mRNA is expressed in some nasopharyngeal carcinoma, suggesting that SCP-1 might be a new tumor target antigen of nasopharyngeal carcinoma.
Adult ; Antigens, Neoplasm ; genetics ; metabolism ; Carcinoma, Squamous Cell ; genetics ; immunology ; metabolism ; Case-Control Studies ; Female ; Humans ; Male ; Nasopharyngeal Neoplasms ; genetics ; immunology ; metabolism ; RNA, Messenger ; genetics ; metabolism

Objective To analyze the efficacy of different doses of intravenous immunoglobulin (IVIG) in the treatment of acquired severe aplastic anemia (AA) in children. Methods The clinical data of hospitalized children with severe AA who received adjuvant immunosuppressive therapy of IVIG from January 2000 to December 2015 were retrospectively analyzed. According to different doses of treatment, the children were divided into low dose group ( IVIG 200-400 mg/ (kg·d) once every 4 weeks for 6 times), high dose group (IVIG 1 g/ (kg·d ) x 2 days once every 4 weeks for 6 times). Results All the children were followed up until December 31, 2015. Among the 61 children, it was effective in 41 children and total effective rate was 67.2%. The effective rate of anti thymocyte globulin (ATG) treatment in high dose group was higher after 3 months than that of low dose group, and there was statistical difference (P=0.020). The interval between first dose of IVIG and first dose of ATG in 20 cases of ineffectiveness was 2.0 (2.0-5.0) d, while that in 41 cases of effectiveness was 8.0 (7.0-9.0) d, and the difference is statistically significant (P<0.001); Among the 20 ineffective children, 18 children had the interval <7 day. The survival rates of the two groups were 80% and 87.1%, respectively, and there was no difference between two groups (P>0.05). The incidence of severe infections in the high-dose group was lower than that in the low-dose group after the use of ATG for 6 months, and there was statistical difference (P=0.008). Conclusions High dose of IVIG therapy can increase the early response rate in children with acquired severe AA, but it does not increase the long-term effectiveness, cure rate and 5 year survival rate. In addition, it can reduce the severe infection rate, but cannot reduce the total infection rate and infection related mortality rate.

Objective To evaluate the effect of different kind of intravenous immunoglobulin (IVIG) therapy in treating Kawasaki disease (KD) and preventing cardiac consequences (coronary artery lesion, CAL). Methods A questionnaire form and guideline for KD diagnosis were sent to 50 hospitals providing pediatric medical care in Shanghai. The data from a total of 1 682 KD patients were collected. It included 1 064 males and 618 females from 1998 through 2008 in Shanghai. The average age of the KD patients was (2.57±2.33) years old (0.1-18.8 years).The patients had been divided into 6 groups for different IVIG therapy, which included 1 g/kg once, 2 g/kg once, 0.4-0.5 g/kg five times, 1 g/kg twice, 2 g/kg twice and others. SAS 6.12 software was used for statistical analysis. Results In all KD patients, the patients treated with IV1G in 5th-10th day of illness has the least cardiac complication and CAL incidence and the group with IVIG therapy of 1 g/kg twice also has the least cardiac complication and CAL incidence. Conclusions The best doses of IVIG in treating KD is 1 g/kg twice and the IVIG therapy should be used in 5th-10th day of KD illness.

ObjectiveTo investigate the effect of Stemona tuberosa alkaloids （STA） on apoptosis and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） and c-Jun N-terminal kinase/p38 mitogen-activated protein kinase （JNK/p38 MAPK） signaling pathways in human lung cancer A549 cells. MethodA549 cells were classified into blank group and STA groups （100， 150， 200， 250， 300 mg⋅L^-1）. Thiazole blue （MTT） assay and colony formation assay were used to evaluate the proliferation of A549 cells. Apoptosis was observed based on Hoechst 33258 staining， flow cytometry， and Annexin V-FITC/PI staining. Western blot was employed to detect the expression of apoptosis-related proteins cysteine-aspartic acid protease-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， and Bcl-2， and the expression of PI3K， phosphorylated （p）-PI3K， Akt， p-Akt， JNK， p-JNK， p38 MAPK， and p-p38 MAPK. ResultCompared with the blank group， STA groups （150， 200， 250， 300 mg⋅L^-1） demonstrated the increase in inhibition rate of cell proliferation （P<0.01） and cell clone inhibition rate， and decrease in cell clone formation rate （P<0.01）. In comparison with the blank group， STA groups showed typical characteristics of apoptosis， such as chromatin condensation and enhanced fluorescence reaction. The apoptosis rate of STA groups was significantly higher than that of the blank group （P<0.01）. Compared with the blank group， STA （150， 200， 250， 300 mg⋅L^-1） significantly up-regulated the protein expression of Caspase-3 and Bax （P<0.05， P<0.01） and down-regulated the expression of Bcl-2 protein （P<0.01）. Compared with the blank group， STA had no significant influence on the total protein expression of PI3K， Akt， JNK， and p38 MAPK. However， STA （150， 200， 250， 300 mg⋅L^-1） significantly decreased the levels of p-PI3K and p-Akt （P<0.05， P<0.01） and increased the level of p-p38 MAPK （P<0.05， P<0.01）. Compared with the blank group， STA （200， 250， 300 mg⋅L^-1） significantly raised the level of p-JNK （P<0.05， P<0.01）. ConclusionSTA can inhibit the proliferation and induce the apoptosis of A549 cells by inhibiting PI3K/Akt signaling pathway and activating JNK/p38 MAPK signaling pathway.

Objective : To investigate the effects of DNA demethylation drugs combined with histone deacetylase inhibitors on fragile X mental retardation 1 neighbor protein (FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression . Methods: Human oral cancer cell lines C al27 and SCC⁃9 were treated with decitabine (DAC) , an inhibitor of DNA methyltransferase , combined with trichostatin A ( TSA) and valproic acid ( VPA) , inhibitors of histone deacetylase . Then reverse transcription⁃polymerase chain reaction ( RT⁃PCR) , quantitative real ⁃time PCR ( qRT⁃PCR) and Western blot were used to detect the expression of FMR1NB and pyrosequencing was used to detect the methylation of FMR1NB promoter. Results : Compared with the blank control group , DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in C al27 and SCC⁃9 cells . Compared with DAC alone group , FMR1NB mRNA expression of each DAC⁃combined drug groups significantly increased , but FMR1NB protein did not significantly change in C al27 cells; for SCC⁃9 cells , except for DAC + TSA group , the mRNA and protein levels of FMR1NB significantly increased in all other groups . In addition , there was no significant difference in the expression of FMR1NB mRNA and protein between the three⁃combined drugs group and two-combined drugs groups . Further methylation assay showed that the methylation level of the overall FMR1NB promoter and its each CpG site measured were reduced to varying degrees in all treatment groups except for three⁃combination drug group of SCC⁃9 . Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter , wherein the enhanced expression effect of the combination of the two drugs is stronger , suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.