Objective To investigate the effects of keratinocyte growth factor(KGF) and keratinocyte growth factor receptor(KGFR) on the malignant transformation of gestational trophoblastic disease(GTD).Methods Immunolocalization of KGF/KGFR was performed on sections prepared with the samples from 26 hydatidiform mole,18 invasive mole and 12 choriocarcinoma.The in situ hybridization was used to detect the mRNA of KGF/KGFR in the tissues of hydatidiform mole and GTD.Analysis was performed according to intensity of staining and number of positive cells.Results It was revealed that specific staining for mRNA and protein of KGF/KGFR existed in hydatidiform mole and gestational trophoblastic tumor(GTT).The mRNA and protein of KGF/KGFR were allocated in cytoplasm of syncytiotrophoblasts and cytotrophoblasts of malignant hydatidiform mole,and the KGF/KGFR protein was also expressed in benign tissue,while the expression of KGFR in malignant hydatidiform mole was significantly higher than that in benign tissue(?2=12.775,P

OBJECTIVE To establish an in vitro test method and to evaluate the genotoxicity of chemicals using primary cultured mouse hepatocytes and the changes in phosphorylated histone H2AX(γH2AX)expression levels to provide a more reliable marker of the identification of genotoxicity. METHODS Hepatocytes were isolated from BALB/c mice by an improved two-step collagenase diges?tion method and then cultured in sandwich configuration. The primary cultured hepatocytes were treat?ed with various concentrations of four known genotoxic agents bleomycin(BLM),benzo(a)pyrene〔B (a)p〕,styrene and styrene-7,8-oxide(SO)within the range of 40 μmol · L-1 and two non-genotoxic agents azathioprine(Aza)and ciclosporin A(CsA)at different time points within 24 h. The cytotoxicity induced by these toxicants was assessed by CCK-8 assay. Then,the changes in γH2AX expression levels in treated cells were determined by flow cytometry. RESULTS The four genotoxic agents could be detected and two non-genotoxic agents could not be detected by this method. The γH2AX expression level was the highest when hepatocytes were exposed to BLM and SO for 3 h,or B(a)p and styrene for 6 h(P<0.01). The production of γH2AX was 25.67,18.36,12.43 and 14.25 for the four types of genotoxic agents,respectively,and was approximately 19,13,9 and 11 times that of the vehicle control group(P<0.01)at the optimum time point and concentration. There was a significant positive corre?lation between the indicated concentrations of genotoxic chemicals and γH2AX expression levels(P<0.01). In addition,the production ofγH2AX indicated no marked increase in two non-genotoxic agents such as Aza and CsA in comparison with the control group. CONCLUSION This test method can effec?tively distinguish genotoxic agents from non-genotoxic agents,and direct genotoxic agents from indirect genotoxic agents in the absence of S9. γH2AX might be a reliable marker for the identification of the potential genotoxicity of chemicals.

Objective To explore and compare the clinical effects of pregnant women with liver cirrhosis after hepatitis in cesarean section between general anesthesia and single spinal anesthesia.Methods Clinical data of pregnant women of 35,including compensated cirrhosis 26 cases,decompensated cirrhosis 9 cases,aged 24-45 years,weighing 55-98 kg,falling into ASA Ⅱ or Ⅲ,were divided into group A (general anesthesia group,n=15) and B (single spinal group,n=20).In patients of group A,propofol were infused at 1.5 mg/kg till the time for fetal delivery,after intubation,followed by infusion of remifentanil at a rate of 0.2 μg·kg-1·min-1 TCL combined with propofol until the end of operation.In group B,the puncture was performed at L3-4 interspace,following by intrathecal injection of ropivacaine 10~15 mg and within 10 seconds.With liver cirrhosis after hepatitis were collected and analyzed,retrospectively.Preoperative general status,blood loss and time of operation,Apgar score of neonates compared and analyzed in groups A and B.And the changes of hepatic function and coagulation function of the body under different anesthesia methods were also studied.The influential factors of postoperative elevation of hepatic function were analyzed by non-conditional logistic regression models.Results Blood loss and time of operation,Apgar score of neonates and postoperative hospital stay had no significant differences between the two groups.Postoperative albumin of group A was higher than that of preoperative,with a statistical significance (P<0.05).Alanine aminotransferase,aspartate aminotransferase of postoperative of group B were increased,with a statistical significance (P<0.01).Logistic analysis showed that the type of anesthesia was the possible influencing factor of postoperative elevation of total bilirubin (OR=12.04,95%CI 1.14-127.45).Conclusion The negative effect of single spinal anesthesia on hepatic function more than that of general anesthesia in pregnant women with liver cirrhosis in cesarean section.

Aim To isolate and characterize the human circulating fibrocytes from human peripheral blood and explore the effects of curcumin on human circulating fi-brocytes.Methods The cells were isolated and puri-fied by density gradient centrifugation,and identified by flow cytometry and immunocytochemistry .Then , CCK-8 and flow cytometry were used to study the effect of curcumin on the proliferation as well as COL I ex-pression of human circulating fibrocytes,respectively. Results After being isolated the cells expressed CD34,CD45 and COLⅠ,among which 79.7% were both CD45 and collagen I positive,typical of human circulating fibrocytes.Curcumin could exert regulatory effects on proliferation of human circulating fibrocytes. Exposure of the cells to curcumin for as short as 24 hours promoted their growth,while prolonged treatment (72 h ) significantly inhibited cell propagation and downregulated the COLⅠ levels,best manifested at a concentration as high as 20 μmol · L-1 .Conclusion The proliferation of cells and COLⅠexpressions can be effectively inhibited by curcumin with the prolonged action period and high concentrations.

Objective To explore the correlation of the myeloid antigen expression and clinical characteristics of acute lymphoblastic leukemia (ALL) in children.Methods The clinical data of 77 newly diagnosed ALL patients in Department of Hematology,the People's Hospital of Zhengzhou University from Jan.2010 to Dec.2013 were analyzed.The patients included 53 boys and 24 girls with a median age of 7.73 (2.00-15.00) years old.Based on flow cytometry (FCM) analysis of bone marrow,these patients were divided into 2 groups:one group included 26 patients with positive myeloid antigen expression (MyAg + ALL) and the other group included 51 patients with negative myeloid antigen expressions (MyAg-ALL).The correlation among myeloid antigen expression,clinical features,prednisone experiment,myelogram on the 15th day was analyzed through induction chemotherapy and minimal residual disease (MRD) on the 33rd day,and the rate of disease-free survival (DFS) was compared between the 2 groups.Results There were 26 cases with myeloid antigen expression among 77 patients (33.77％),CD13 + accounting for 19.48％ (15/77 cases),CD33 + 10.39％ (8/77 cases),and CD117 + 5.19％ (4/77 cases).Among these patients,there were 2 patients expressing both CD13 + and CD33 +,and 1 patient expressing both CD33 + and CD117 +.There was no difference between the MyAg + ALL group and MyAg-ALL group in gender (x2 =0.217,P =0.641),age (≥ 10 years old,x2 =0.011,P =0.918),white blood count(≥50 × 109/L,x2 =1.198,P =0.274),lactate dehydrogenase (LDH) (≥500 U/L,x2 =0.317,P =0.573),genetic abnormality (x2 =0.377,P =0.539),immunophenotype (B-ALL/T-ALL,x2 =0.397,P =0.529),and risk stratification (low-risk group,middle-risk group and high-risk group,x2 =0.260,P =0.878).Univariate Logistic regression showed that the reaction rate of prednisone experiment (P =0.023,OR =3.422) and positive rate of MRD (P =0.001,OR =0.133) of MyAg + ALL group were obviously higher than those in MyAg-ALL group.Multivariate Logistic regression showed that positive rate of MRD in CD13 + ALL group was obviously higher than that of CD13-ALL group (P =0.034,OR =120.765).The DFS rate of CD13 + ALL group and CD13-ALL group were (50.4 ± 13.8)％ and (77.4 ±6.7)％ respectively,and there was a significant difference between the 2 groups (x2 =3.928,P =0.047).Conclusions There is no significant correlation between myeloid antigen expression and clinical characteristics of children patients with ALL.For the patients with myeloid antigens,the early reaction of induction chemotherapy is bad,and for patients with CD13,the prognosis is not good.

Objective To evaluate the diagnostic value of six-slice coronal reformations in patients with acute midepigastric pain. Methods A total of 974 patients with acute midepigastric pain were included in this study and divided into group A(coronal reformation)and group B(non-coronal reformation).For group A,reconstructed coronal and oblique-coronal images were acquired.A comprehensive diagnosis was made based on coronal reformations and axial planes.Anatomical nomenclature was adopted,including kidney-ureter plane,abdominal aorta plane,superior mesenteric artery plane,ascending colon-appendix plane,stomach-cholecyst plane and colon-small intestine plane.For group B,the diagnosis was made based on axial planes.Finally,a comprehensive analysis was made,missed cases in these two groups were counted and compared,and statistical analysis was performed using the SPSS software(version SPSS V17).Results For group A,the missed diagnosis was made in 12 cases(1.23%)and it was 53 cases(6.58%)for group B.There was a statistically significant difference between two groups(P<0.05).Conclusion Combined with axial planes or oblique coronal reformations,six-sclice coronal reformation can reduce the the rate of missed diagnosis of acute midepigastric pain.

Objective: To investigate the prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric patients with t (8;21) acute myeloid leukemia (AML) . Methods: The clinical features and RUNX1-RUNX1T1 transcript levels of 55 pediatric t (8;21) AML patients, newly diagnosed from Jan. 2010 to Apr. 2016, were analyzed retrospectively. The relationship between the minimal residual disease (MRD) and prognosis was analysed by dynamic monitoring of RUNX1-RUNX1T1 transcript levels using real-time quantitative PCR (RQ-PCR) technology. Results: The RUNX1-RUNX1T1 transcript levels in bone marrow cells at diagnosis was not related to relapse. After one course of induction therapy, patients with a more than 2 Log reduction of RUNX1-RUNX1T1 transcript levels (>2 Log) had lower 5 years cumulative incidence of relapse (CIR) [ (24.3±8.4) % vs (52.6±9.7) %, χ²=9.046, P=0.003], relapse-free survival (RFS) [ (71.6±12.7) % vs (48.1±13.2) %, χ²=5.814, P=0.016], and better overall survival (OS) [ (76.9±12.5) % vs (48.9±14.7) %, χ²=6.346, P=0.012], compared to patients with a less than 2 Log reduction (a<2 Log) . Multivariate Cox survival analysis suggested that a>2 Log reduction in RUNX1-RUNX1T1 transcript levels after a course of induction therapy was an independent prognostic factor for RFS (HR=0.263, 95%CI 0.081-0.851, P=0.026) and OS (HR=0.214, 95% CI 0.057-0.808, P=0.023) . During consolidation therapy and follow-up period, molecular relapse of 16 cases and hematologic relapse of 13 cases were identified by continuous dynamic monitoring of RUNX1-RUNX1T1 transcript levels, with a median interval of 4.0 (1.5-5.8) months from the molecular relapse to hematologic relapse. 2 cases of molecular relapse who received timely allogeneic hematopoietic stem cell transplantation did not experience hematologic relapse. Conclusion Dynamic monitoring RUNX1-RUNX1T1 transcript levels by RQ-PCR technique can subdivide patients into relatively low and high risk group, early screen patients at high risk of relapse and provide a scientific basis for precision stratification and risk-adapted therapy for pediatric t (8;21) AML children.

Humans ; Interleukin-16 ; immunology ; Multiple Myeloma ; immunology

Objective:To investigate the role of IL-21/IL-21R-mediated CD4 + T cells in Chlamydia muridarum ( Cm) respiratory infection. Methods:C57BL/6 mice (WT mice) and IL-21R -/- mice were used to establish the models of Cm respiratory infection through intranasal inhalation of Cm. Flow cytometry was used to detect the proportion, number, activity and function of CD4 + T cells in lung and spleen tissues at 0, 3, 7 and 14 d after Cm respiratory tract infection. IFN-γ and IL-4 levels in spleen cell culture supernatants were detected by ELISA. Na?ve WT mice were transferred with CD4 + T cells in the spleen tissues of IL-21R -/- mice or WT mice on 7 d after infection and given Cm intranasally 2 h later. Then the mice were weighed daily and sacrificed on 14 d after infection. The bacterial load and pathological changes in lung were analyzed. Flow cytometry was performed to detect the proportions and numbers of neutrophils (CD45 + CD11b + Gr-1 high) and alveolar macrophages (CD45 + F4/80 + CD11c high)as well as the proportions of Th1 (IFN-γ + CD4 + ) and Th2 (IL-4 + CD4 + ) cells. ELISA was also performed to measure IFN-γ and IL-4 levels in spleen cell culture supernatants. Results:Compared with WT mice, IL-21R -/- mice showed elevated numbers and enhanced activation of CD4 + T cells, increased proportion of Th1 cells and decreased proportion of Th2 cells in spleen and lung tissues after Cm respiratory infection. Besides, IFN-γ levels increased, while IL-4 levels decreased in spleen cell culture supernatants of IL-21R -/- mice. After Cm infection, the na?ve WT transferred with CD4 + T cells from IL-21R -/- mice showed less body weight loss, reduced bacterial load and alleviated pathological changes in lung tissues, increased proportion of Th1 cells in lung tissue and higher IFN-γ level in spleen cell culture supernatants. Conclusions:IL-21/IL-21R-mediated CD4 + T cells could aggravate Cm respiratory infection by suppressing Th1 cell immune responses.

OBJECTIVE: To establish a PC12 cell line with stable expression of human apolipoprotein E( ApoE4) gene by transfection with a lentiviral vector carrying human ApoE4 gene and to investigate the effect of maltol aluminum on the viability of transfected PC12 cells. METHODS: The lentiviral vector carrying human ApoE4 gene was transfected into PC12 cells. PC12 cells with overexpression of ApoE4 gene and negative control vector were obtained after puromycin screening.The mRNA relative expression of R-Apo E and( or) H-Apo E-FLAG of cells in PC12,PC12-NC and PC12-ApoE4 groups were detected by real-time fluorescent quantitative polymerase chain reaction,and the effect of cell construction was identified. PC12-ApoE4 cells and PC12 cells were exposed to maltol aluminum solution at concentrations of 0. 00,100. 00,200. 00 and 400. 00 μmol/L respectively for 24 hours,and cell viability was detected by Cell Counting Kit-8( CCK-8)assay. RESULTS: PC12-ApoE4 and PC12-NC cells under the fluorescence microscope showed fluorescence expression,suggesting that transfection was successful. The expression of PC12 cells showed no fluorescence. The relative expression of H-Apo E-FLAG gene mRNA( the median amount) of PC12-ApoE4 cells was 148. 74,which was higher than the R-Apo E gene in PC12 cells( 1. 00) and PC12-NC cells( 1. 01)( P < 0. 01). After exposure to maltol aluminum,the cell survival rates in terms of the main effect and interaction effect of dose and cell type were statistically significant( P < 0. 01),among them,the cell viabilities were decreased in the concentration range of 0. 00-400. 00 μmol/L with the dose of maltol aluminum exposure increased,showing dose-effect relationship( P < 0. 01). CONCLUSION: The cell line stably expressed human ApoE4 gene was constructed successfully. There was interaction between the effects of maltol aluminum and ApoE4 gene on the survival rate of PC12 cells,and ApoE4 gene could enhance the cytotoxicity of maltol aluminum on PC12 cells.