Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

OBJECTIVETo explore the diagnosis and treatment of pregnancy-associated acute Stanford type A aortic dissection to improve the maternal and fetal outcomes.

METHODSWe analyzed the perioperative data of 5 pregnant women with acute Stanford type A aortic dissection treated between June, 2009 and February, 2017.

RESULTSThe median age of the women was 30 years (range, 22-34 years) with gestational weeks of 23-38 weeks upon diagnosis. All the 5 patients received surgical interventions. Three patients underwent caesarean delivery and hysterectomy, and the fetuses survived after the surgery; 2 patients chose to continue pregnancy following the surgery, among whom one died due to postoperative complications and the other underwent termination of pregnancy. During follow-up, the surviving patients showed no endoleak in the descending aorta stent and the distal dissection remained stable.

CONCLUSIONThe maternal and fetal outcomes of pregnancy-associated acute Stanford type A aortic dissection can be improved by multidisciplinary cooperation and optimization of the surgical approaches according to the time of pregnancy, fetal development and conditions of the aortic lesions.

OBJECTIVETo explore the inhibitory effect of estrogen against metastasis of human hepatocellular carcinoma MHCC97H cells and explore the molecular mechanism.

METHODSThe inhibitory effect of estrogen on the migration and invasion of MHCC97H cells was evaluated with wound healing assay and Transwell assay. Western blotting was used for investigating the expression of MMP-2, MMP-9, AKT and p-AKT in the cells treated with estrogen.

RESULTSEstrogen treatment significantly inhibited the migration and invasion of MHCC97H cells in a dose-dependent manner. Estrogen significantly down-regulated the protein expressions of MMP-2 and MMP-9 and lowered the phosphorylation level of AKT.

CONCLUSIONThe anti-metastatic effect of estrogen involves inhibition of MMP-2 and MMP-9 in MHCC97H cells probably by regulating AKT signal pathway.

Objective To investigate the effect of inhibiting S100A4 expression to enhance the dissociation of mast cells pre-bound by immunoglobulin E(IgE)by omalizumab(OmAb).Methods LAD2 cells were randomly divided into normal group,IgE group(IgE induction),OmAb-L group(0.5 mg·mL-1 OmAb),OmAb-M group(1.0 mg·mL-1 OmAb),OmAb-H group(2.0 mg·mL-1 OmAb),OmAb-h+si-S100A4 group(transfected with si-S100A4+2.0 mg·mL-1 OmAb).IgE levels on cell surface were detected by flow cytometry;degranulation was measured by β-amino-hexosidase release assay;the levels of histamine and leukotriene C4 were detected by enzyme-linked immunosorbent assay(ELISA);the expression of related proteins was detected by Western blot.Results After 6 h treatment,IgE levels in normal group,IgE group,OmAb-H group and OmAb-H+si-S100A4 group were(4.13±0.52)％,(100.00±6.20)％,(60.12±3.41)％and(54.04±5.60)％,respectively;β-amino-hexosidase release rates were(12.59±1.35),(69.27±6.43),(45.39±2.14)and(37.80±2.77)％,respectively;histamine levels were(2.43±0.16),(8.57±0.41),(4.91±0.24),(3.01±0.23)ng·mL-1,respectively;the C4 levels of leukotriene were(198.85±18.91),(423.56±1.25),(273.68±17.11)and(242.79±12.44)pg·mL-1,respectively;relative phosphorylated extracellular signal-regulated kinases(p-ERK)expression levels were 0.31±0.04,0.91±0.12,0.55±0.04 and 0.35±0.02,respectively.The above indexes in IgE group were compared with those in normal group,the above indexes of OmAb-H group were compared with IgE group,the above indexes of OmAb-H+si-S100A4 group were respectively compared with those of OmAb-H group,the differences were statistically significant(all P＜0.05).Conclusion Inhibition of S100A4 can enhance the dissociation effect of OmAb on mast cells and IgE,and further block the release of allergic mediators.

OBJECTIVETo investigate the inhibitory effect of resveratrol on interleukin-1β (IL-1β) production in mesenchymal stem cells (MSCs) exposed to radiation and the action mechanism of resveratrol.

METHODSMSCs were divided into blank control group, radiation group, shRNA interference group, and resveratrol groups. The resveratrol groups were given different doses of resveratrol (50, 100, and 200 µmol/L) before radiation. The secretion and expression of IL-1β was measured by enzyme-linked immunosorbent assay, Western blot, and RT-PCR.

RESULTSCompared with the radiation group, the resveratrol groups had significantly decreased extracellular secretion of IL-1β (t = 83.34, 24.48, and 12.52, P < 0.05 for all) and significantly decreased intracellular expression of IL-1β protein and mRNA (t = 8.695, 14.77, and 13.9, P < 0.05 for all). Compared with those given 200 µmol/L resveratrol alone before radiation, the MSCs treated by SIRT1 silencing and given 200 µmol/L resveratrol before radiation had significantly increased extracellular secretion of IL-1β (t = 18.57, P < 0.05) and significantly increased intracellular expression of IL-1β protein and mRNA (t = 10.24, P < 0.05).

CONCLUSIONResveratrol can significantly inhibit the production of IL-1β in MSCs exposed to radiation, and SIRT1 may play a key regulatory role in the process of inflammation induced by radiation.

Cells, Cultured ; Humans ; Interleukin-1beta ; metabolism ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; radiation effects ; Radiation ; Radiation Dosage ; Stilbenes ; pharmacology

Objective:The goal of this study was to analyze the clinical significance of relationship between myocardial collateral and the levels of hypoxia‐inducible factor 1‐alpha (HIF‐1α) and vascular endothelial growth factor A (VEGF‐A) in patients with coronary chronic total occlusion lesion .Methods:89 patients with coronary chronic total occlusion lesion confirmed by clin‐ical data and coronary angiography were identified .The levels of HIF‐1αand VEGF‐A were measured by ELISA ,and the rela‐tive expression of VEGF‐A of peripheral blood mononuclear cell (PBMC) were measured by real‐time PCR .The results were statistically analyzed by the statistical programme for social sciences (SPSS version 18 .0) and software SAS JMP 9 .0 .Results:Compared to Rentrop 0‐1 grade group (18/38 ,47 .4% ) ,Rentrop 2 (11/31 ,35 .5% ) and Rentrop 3 (3/20 ,15 .0% ) grade group had fewer diabetes mellitus .Rentrop 2 [(6 .67 ± 1 .41) mmol/L] and Rentrop 3 [(5 .48 ± 1 .26) mmol/L] grade group had low‐er fasting blood glucose than Rentrop 0‐1 grade group [(7 .24 ± 1 .39) mmol/L] .Rentrop 2 (12/31 ,38 .7% ) and Rentrop 3 (3/20 ,15 .0% ) grade group had fewer clinical heart failure (NYHA Ⅱ ~ Ⅳ grade) than Rentrop 0‐1 grade group (20/38 , 52 .6% ) .Rentrop 2 [(85 .5 ± 27 .7) pg/mL ,(139 .5 ± 42 .1) pg/mL] and Rentrop 3 [(103 .3 ± 30 .2) pg/mL ,(162 .6 ± 43 .3) pg/mL] grade group had higher levels of HIF‐1αand VEGF‐A than Rentrop 0‐1 grade group [(42 .0 ± 16 .1) pg/mL ,(76 .5 ± 32 .2) pg/mL] .Rentrop 2 (1 .31 ± 0 .46) and Rentrop 3 (1 .38 ± 0 .44) grade group had higher level of relative expression of VEGF‐A in PBMC than Rentrop 0‐1 grade group (1 .00 ± 0 .28) .Conclusions:Chronic and consistent ischemia and hypoxia in‐duced the increase of expression of HIF‐1αand VEGF‐A is important for establishment of coronary collateral ,increasing blood supply and improving the heart function and prognosis .

OBJECTIVEThis study was to investigate the cell morphology and cell immune phenotypic characteristics in patients with multiple myeloma (MM).

METHODSThe flow cytometry with multiparametric direct immunofluorescence technique, and CD45/SSC and CD38(+)(+)/CD138(+) gating were used to measure cell markers CD138, CD38, CD56, CD117, CD3, CD13, CD33, CD19, CD7, CD20, CD22, CD34, CD28 in 47 MM patients. At the same time the morphology examination of bone marrow cells was performed.

RESULTSThe suspicious myeloma cell ratio in MM patients was 9.42%-74.25% detected by flow cytometry, moreover, the myeloma cell ratio detected by morphology examination was 11.0%-80.6%, there was a good correlation between the two detection methods (r(2) = 0.54, P < 0.001). The ratio of antigen positive expression was as follows: 74.46% for CD138, 100% for CD38, 57.44% for CD56, 40.42% for CD117, 6.38% for CD13, 19.15% for CD33, 8.51% for CD20, 27.66% for CD28, 2.12% for CD22, 4.25% for CD34, 0% for CD3, 0% for CD19, 0% for CD7.

CONCLUSIONSCD45/SSC and CD38(+)/CD138(+) gating technique can accurately gate multiple myeloma cell sets which need analysis, the majority of myeloma cells expreses CD138, CD38, CD56 antigens. The immunophenotypic analysis combined with the cell morphology examination more contribute to the diagnosis and differential diagnosis of multiple myeloma.

Antigens, CD ; Bone Marrow Cells ; Flow Cytometry ; Humans ; Immunophenotyping ; Multiple Myeloma

The aim of this study was to detect the mRNA expression of tissue factor pathway inhibitor-2 ( TFPI-2) and its methylation in bone marrow mononuclear cells from acute myeloid leukemia (AML) patients and to explore its significance in AML. Bone marrow mononuclear cells were isolated from newly diagnosed AML patients (n = 33), complete remission AML patients (n = 19), relapsed/refractory AML patients (n = 12) and iron deficiency anemia patients (control group, n = 15). Expression of TFPI-2 mRNA was detected with real-time quantitative PCR (RT-PCR) and the methylation of CpG island in its promoter was detected with methylation-specific PCR (MSP). The results showed that the expression of TFPI-2 mRNA in newly diagnosed AML, complete remission AML and relapsed/refractory AML patients was much lower than that in the controls (P < 0.05). Furthermore, its expression in relapsed/refractory AML patients was lower than that in newly diagnosed AML patients (P = 0.006). Compared with complete remission AML patients, the expression of TFPI-2 mRNA in newly diagnosed AML patients was significantly reduced (P = 0.030). The percentage of TFPI-2 promoter methylation in AML patients was 64.63% (42/64). In newly diagnosed AML group, complete remission AML group and relapsed/refractory AML group,the percentages of TFPI-2 promoter methylation were 66.67% (22/33), 52.63% (10/19) and 83.33% (10/12) (P > 0.05), respectively. The optical density ratio of TFPI-2 mRNA expression was 0.165 (0.005-2.099) in methylated AML patients, and 0.597 (0.011-2.787) in unmethylated AML patients (P < 0.05). Methylation of TFPI-2 gene promoter was not detected in control patients. After 2 courses of chemotherapy, the level of TFPI-2 mRNA was much higher in the CR group than that in the non-CR group (P < 0.05). It is concluded that the down-regulation or silence of TFPI-2 gene potentially results from its promoter methylation, and the expression level of TFPI-2 and the methylation status of its promoter may be used as indicators of risk stratification and evaluation of disease progress.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; DNA Methylation ; Female ; Glycoproteins ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; pathology ; Male ; Middle Aged ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVE: To compare the treatment outcome and prognosis of the newly-treated myc METHODS: 152 double-expression lymphoma patients (myc RESULTS: The median age of 152 DEL patients was 60.5 years old (15-87 years old). 85 patients (55.9%) were Ann Arbor stage III/IV. There was no significant difference in clinical data between the patients in the two groups. Multivariate Cox regression analysis showed that bcl-6 expression, ECOG score, and stage were the independent prognostic factors for the entire group of DEL patients. There was no statistical difference in ORR between the patients in the two groups (χ2=0.749, P=0.387). Kaplan-Meier survival analysis showed that PFS and OS of the bcl-6 CONCLUSION bcl-6
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; Cyclophosphamide ; Doxorubicin ; Humans ; Lymphoma, Large B-Cell, Diffuse/drug therapy* ; Middle Aged ; Prednisolone ; Prognosis ; Vincristine/therapeutic use* ; Young Adult

OBJECTIVETo ovaluate the prognostic value of prednisone response in treatment regimes of children with acute lymphoblastic leukemia.

METHODSA total of 598 newly diagnosed ALL patients were enrolled and received prednisone pre-treatment. Based on the peripheral lymphoblast count on day 8, these patients were divided into 2 groups: prednisone good response (PGR) and prednisone poor response (PPR). PPR patients were classified into high risk group immediately and then received intensed chemotherapy. The all enrolled patients were followed up and the clinical features and treatment outcomes of the two groups were analyzed.

RESULTSCompared with PGR group, PPR group had different characteristics. They were older in age and had higher initial white blood cell count (P<0.05). T-cell ALL (T-ALL) and Philadelphia chromosome positive ALL (Ph+ ALL) were frequent in PPR group (P<0.05). Event-free survival (EFS) rate of PPR group was significantly lower than that of PGR group (P<0.05). 2 year event-free survival(EFS) rate of PGR group was (88.3±1.5)%, while the 2-year EFS rate of PPR group was (58.4±5.3)%. 5 year EFS rates of PGR and PPR were (80.8±2.1)% and (53.4±6.0)%, respectively. The EFS rate of PPR group was falling rapidly within 2 years. PPR group had higher relapse rate, and most relapses occurred within 18 months (P<0.05). PPR group had more high incidence of minimal residual disease (MRD) both on day 33 and on week 12 (P<0.05). No significant difference of EFS and relapse time was found between PPR and high risk PGR patients (P>0.05). In multi-variate regression analysis, the PPR, the presence of BCR-ABL1 and MLL were significantly unfavorable factors (P<0.05).

CONCLUSIONPrednisone response has been confirmed to be still great prognostic value and PPR children patients have poor outcomes generally. It is likely that the response to prednisone does not make much sense to high risk ALL patients.

Disease-Free Survival ; Humans ; Multivariate Analysis ; Neoplasm, Residual ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prednisone ; Prognosis ; Recurrence ; Treatment Outcome