Down-regulation of ABCB4 expression in placenta inhibits glycocholic acid evacuation in fetus and is related with poor prognosis

AIM: To investigate the relationship between ABCB4 (ATP-binding cassette, sub-family B, member 4) expression in placenta and glycocholic acid (GA) evacuation in fetus for evaluating the prognosis of fetus from women with intrahepatic cholestasis of pregnancy (ICP).METHODS: Normal pregnant women (control group, n=30) and ICP women (ICP group, n=30) hospitalized in our department were selected. The expression of ABCB4 mRNA and protein in placenta was detected by RT-PCR and immunohistochemistry, respectively. The total bilirubin (TBIL) and alanine aminotransferase (ALT) levels in maternal blood and the GA levels in maternal or umbilical blood were measured by chemical oxidation method, enzyme kinetics method and radioimmunoassay, respectively. RESULTS: The expression of ABCB4 mRNA and protein in placenta of ICP patients was significantly lower than that of the controls (P<0.01). The TBIL and ALT levels in maternal blood and the GA levels in both maternal and umbilical blood of ICP women were higher than those of the controls (P<0.01). The GA levels in maternal and umbilical blood were negatively correlated with the ABCB4 mRNA levels in placenta (r=-0.627 and r=-0.795, respectively, P<0.01). In control group, the weight and Apgar scores of the newborns were higher than those in ICP group (P<0.01). However, the rate of stool contamination in amniotic fluid was lower in control group than that in ICP group (P<0.01).CONCLUSION: Down-regulation of ABCB4 expression is involved in the impairment of bile acid secretion in placenta and may be a marker for poor prognosis of the fetus from ICP women.     

Protein and mRNA expression of visfatin in placenta of patients with preeclampsia

AIM: To investigate the expression of visfatin in the placenta of patients with preeclampsia and its significance. METHODS: The pregnant women(n=100) were divided into normal pregnancy group, mild preeclampsia group and severe preeclampsia group according to the severity of the disease. The pathological changes of the placenta were observed by hematoxylin-eosin staining. The expression of visfatin at mRNA and protein levels in the placenta was detected by real-time PCR and immunohistochemistry respectively. RESULTS: Compared with normal pregnancy group, the pathological changes of the placenta in preeclampsia groups was significant, showing that the structure and the form were disordered and incompleted in both cytotrophoblasts and syncytiotrophoblasts. The proliferation of cytotrophoblasts and the numbers of the placental villi with syncytial knots were observed. The vascular numbers of villi were decreased and congested. The results of immunohistochemistry and real-time PCR showed that the visfatin protein was observed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts among 3 groups. The expression of visfatin at mRNA and protein levels was increased with the severity of the preeclampsia. CONCLUSION: The injury and dysfunction of vascular endothelial cells in the villi exist in the patients with preeclampsia. High expression of visfatin at mRNA and protein levels in placenta of the patients with preeclampsia indicates that visfatin is closely related to preeclampsia.     

Estradiol promotes growth of decidua derived mesenchymal stem cells by inhibiting miR-16 via estrogen receptor α

AIM: To study the effect of estradiol (E₂) on the viability of mesenchymal stem cells (MSCs) derived from the decidua of the placenta by regulating the expression of microRNA-16 (miR-16). METHODS: The concentration of E₂ in the peripheral blood of normal pregnant women and the patients with severe preeclampsia (PE) was measured. The effects of E₂ at different concentrations on the viability of MSCs were analyzed. The effect of E₂ at different concentrations on the expression of miR-16 in the MSCs was detected, and which estrogen receptor (ER) mediated the regulatory effect of E₂ on miR-16 expression was determined. RESULTS: The concentration of E₂ in peripheral blood of the patients with severe PE was significantly decreased (P<0.01). After treatment with E₂ at 5, 10 and 100 nmol/L for 48 h, the viability of MSCs was increased (P<0.05). The expression level of miR-16 was down-regulated in the MSCs treated with E₂ at 5, 10 and 100 nmol/L for 12 h. After treatment with E₂ at 10 nmol/L for different time (0 h, 3 h, 6 h, 12 h and 24 h), the expression level of miR-16 in the MSCs showed a clear time-dependent downward trend. E₂ significantly promoted the viability of MSCs, and the cell viability was significantly reversed after miR-16 pretreatment. Pretreatment with estrogen receptor antagonists ICI 182780 and tamoxifen for 6 h attenuated the inhibitory effect of E₂ on miR-16 expression. Only ERα agonist propyl pyrazole triol significantly inhibited the expression of miR-16 in MSCs but ERβ agonist diarylpropionitrile did not. CONCLUSION: E₂ promotes the growth of decidua-derived MSCs by inhibiting miR-16 via ERα.     

Preliminary study of endothelial progenitor cells formed vessel-like structure in vitro

AIM: Endothelial progenitor cells (EPCs) are a group of stem cells/progenitor cells, which exist in postnatal body and can be of specially homing to the foci of angiogenesis and then differentiate into endothelial cells.This investigation was to study the method for culturing endothelia progenitor cells (EPCs) in vitro, and to observe its feasibility and condition formed vessel-like structure.METHODS: The cells were isolated from born marrow, peripheral blood, umbilical cord blood or spleen in different laboratories.The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells under the induction of special cytokines.The expression of CD34, VEGFR-2, AC133 and VE-cadherin were detected by fluorescence-activated cell sorting.The endothelial cell lineage was confirmed by DiI-ac-LDL up-taking and Ⅷ factor immunocy tochemistry.RESULTS: The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro successfully, forming cord-like and tube-like structure.The EPCs derived from rabbit peripheral blood differentiated more mature and formed vessel-like structure.CONCLUSION: The EPCs derived from human umbilical cord blood and rabbit peripheral blood formed vessel-like structure in vitro.EPCs may be a potential resource of vessel tissue engineering.     

Influence on killing effect of NK cells in vitro by up-regulation of HLA-E expression in hepatocellular carcinoma Bel7402 cells

AIM: To investigate the influence on the killing effect of NK cells in vitro by up-regulation of human leukocyte antigen-E (HLA-E) expression in hepatocellular carcinoma Bel7402 cells. METHODS: The recombinant lentiviral vector (Lentivirus/CMV/GFP-HLA-E) was constructed and transfected into hepatocellular carcinoma Bel7402 cells. The HLA-E gene expression at mRNA and protein level was monitored by the methods of real-time RT-PCR and Western blotting. The influence on the killing effect of NK cells in vitro by up-regulation of HLA-E expression in hepatocellular carcinoma Bel7402 cells and by HLA-ABC antibody blocking the site on the surface of target cells was analyzed.RESULTS: Real-time RT-PCR showed that there was a significant increase in HLA-E mRNA level in hepatocellular carcinoma Bel7402 cells transfected with Lentivirus/CMV/GFP-HLA-E at 24 h (P<0.05) and 48 h,72 h, 96 h (P<0.01) as compared to blank group. There was also a significant increase in exogenous/endogenous HLA-E proteins at 12 h, 24 h, 48 h, 72 h and 96 h by Western blotting (P<0.01). Without HLA-ABC antibody blocking, there was a statistical difference for the killing effect of NK cells,comparing Bel7402 Lenti HLA-E group with Bel7402 group (P<0.05). Comparing HLA-ABC antibody blocking group with no HLA-ABC antibody blocking group, a statistical difference for the killing effect of NK cells (P<0.05) was observed. There was also a statistical difference for the killing effect of NK cells between Bel7402 with blocking group and Bel7402 Lenti HLA-E with blocking group (P<0.05). CONCLUSION: The vector of Lentivirus/CMV/GFP-HLA-E has an active up-regulation effect in HLA-E mRNA level and HLA-E protein level. While up-regulation of HLA-E in target cells, the killing effect of NK cells on target cells is obviously weakened. Blockage of the sites on the surface of target cells by HLA-ABC antibody universally enhances the killing effect of NK cells on the target cells.     

Percentage change on natural killer subsets of peripheral blood and decidua in between normal early pregnancy and spontaneous abortion

AIM: To determinate the percentage change on natural killer(NK) subsets of peripheral blood and decidua in between normal early pregnancy and spontaneous abortion. METHODS: Flow cytometry was used to detect the expression of CD56 and CD16 on lymphocytes of decidua and peripheral blood in normal early pregnancy and spontaneous abortion. RESULTS: In peripheral blood, the percentage of CD56⁺ of spontaneous abortion tended to decrease in comparison with normal early pregnancy, and the percentage of CD56⁺CD16⁺ in spontaneous abortion was significantly less than that in normal early pregnancy. The percentages of different natural killer subsets in decidua of spontaneous abortion were significantly lower than those in decidua of normal early pregnancy. CONCLUSION: The decrease of CD56⁺NK cells in decidua may be one of causes for abortion, the loss of CD56⁺ and CD56⁺CD16^+NKcells in peripheral blood could have a diagnostic value for spontaneous abortion.     

Serum levels of visfatin and tumor necrosis factor-alpha in patients with pre-eclampsia and their relationship with insulin resistance

AIM: To explore the serum levels of visfatin (VF) and tumor necrosis factor-alpha (TNF-α) in the patients with pre-eclampsia (PE) and their correlation with insulin resistance (IR). METHODS: The severe PE patients (n=30), mild PE patients (n=30) and normal pregnant women (n=40) were selected according to the classification standard of PE. The serum levels of VF and TNF-α were measured by ELISA. Fasting plasma glucose (FPG) and fasting insulin (FIns) were detected by glucose oxidase method and radioimmunoassay, respectively. Triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by an automatic biochemical analyzer. According to calculating the mean arterial pressure (MAP), body mass index (BMI) and homeostatic model assessment for insulin resistance index (HOMA-IR), the correlation between IR and the levels of serum VF as well as TNF-α were analyzed.RESULTS: The levels of VF and TNF-α in severe PE group and mild PE group were significantly lower than those in normal pregnancy group (P<0.05). In addition, the levels of VF and TNF-α in severe PE group were lower than those in mild PE group (P<0.05). Linear correlation analysis showed that serum VF was positively correlated with TNF-α and HDL-C (P<0.05), and negatively with MAP and FIns (P<0.05). The serum TNF-α was positively correlated with HDL-C (P<0.05), and negatively with BMI, TG, MAP and FIns (P<0.05). Multiple stepwise regression analysis showed that FBG, FIns and HOMA-IR were relative independent factors of serum VF and TNF-α (P<0.05).CONCLUSION: Serum levels of VF and TNF-α are closely related to IR.     

Role of NK cells in mouse allogeneic bone marrow transplantation

AIM: To study the effect of natural killer(NK)-cells on graft-versus-host disease(GVHD), graft rejection, engraftment and reconstituting of hematopoiesis in mouse allogeneic bone marrow transplantation. METHODS: Lethally irradiated BALB/c(H-2^d) mice were transplanted with C57/6j(H-2^b) bone marrow containing donor peripheral T cells and/or NK cells. Recipients CD₃₄⁺ cell counts and the expression of H-2K^b＋ cell were detected by flow cytometry, peripheral white blood cell(WBC) was detected by auto-cytometry, and the recipients survival rates, GVHD, engraftment and hematopoiesis recovery were then observed. RESULTS: In the group of transplanted with NK cells infusion, the incidence of GVHD was evidently lessened, the survival rates, WBC and CD₃₄⁺ cell counts and the expression of H-2K^b＋ cell were significantly high than that without NK cells infusion. CONCLUSION: In mouse allogeneic bone marrow transplantation, alloreactivity NK cells prevents GVHD, reduces graft rejection, and promotes engraftment and reconstituting of hematopoiesis.     

Transfusion of mesenchymal stem cells improve the hematopoiesis in mice with aplastic anemia by regulating immune cells

AIM: To study the possible immunologic mechanism of mesenchymal stem cells (MSCs) by which the hematopoiesis of mice with aplastic anemia (AA) is improved. METHODS: The mice model of immuno-mediated aplastic anemia was established. Thirty BALB/c mice were divided into 3 groups: radiation group, AA model group and MSCs group. The mice in radiation group only had processing of 5 Gy ［60Co］γ radiation. The mice in AA model group were intravenously injected with the cell suspension of thymocyte and lymph-node cell mixture (prepared from DBA/2 mice, 1×106 cells) after 5 Gy ［60Co］γ radiation. The animals in MSCs group received the same treatment as the mice in AA model group did, and afterwards was injected with 1×106 MSCs intravenously at 3 days. All the mice in the 3 groups were observed and analyzed the following parameters: peripheral blood cells, pathological features of bone marrow, the relation between the changes of peripheral CD4+, CD8+, CD4+/CD8+, NK cells and the hematopoiesis. RESULTS: After 5 Gy ［60Co］γ radiation, the peripheral blood cells in all groups decreased at 7 days, while at 21 days those in MSCs group and radiation group restored to normal but those in AA model group was still at lower level. At 7 days, the cells of CD4+, CD8+ and CD4+/CD8+ in each group were similar without statistical difference, but the number of NK cells in MSCs group was quite lower than that in the other 2 groups (P<0.05). At 14 days, the CD4+ cells in all groups were obviously decreased, while the CD8+ cells in MSCs group and AA model group were significantly higher than those in radiation group. At 21 days, the CD8+ and NK cells in MSCs group were quite the same as those in radiation group while those in AA model group were obviously higher compared to the other 2 groups. The number of adipose cells observed in the pathological slice of femoral bone in MSCs group and radiation group was no difference while that in AA model group was much higher. CONCLUSION: MSCs transfusion improves the hematopoiesis in mice with aplastic anemia by regulating the functions of immune cells.     

Effect of pregnancy-associated glycoproteins on E-selectin-medi ated endothelial cell adhesion

AIM:To explore the effect of high purified human chorionic gonadotropin (hCGs) and glycodelins isolated from amniotic flui d and serum of pregnant women on E-selectin mediated endothelial cell adhesion.METHODS:The hCGs and glycodelins in human amniotic fluid and se rum collected from pregnant women were purified by using chromatography.The bin ding of[51Cr］-labelled HepG2 cells to IL-1β stimutated human umbilical vein endothelial cells (HUVECs) in the presence of increasing concentrations of hCGs and glycodelins was quantified.The effect of hCG and glycodelin on E-selec tin-mediated endothelial cell adhesion was observed.RESULTS:HCGs isolated from amniotic fluid and serum of pregnant women inhibited the E-selectin mediated adhesion of HepG2 cells to HUVECs in a dose-dependent manner,the IC₅₀ was 3.2×10-6 mol/L and 6.3×10 -6 mol/L,respectively and the relative inhibition was 1.5×104 and 1 .1×104,respectively,compared to sialyl-Lewisx.The IC₅₀ of amnioti c fluid glycodelin and serum glycodelin was 3.2×10-6 mol/L and 6.3×10 -6 mol/L respectively,and the relative inhibition was 7.2×102 and 3.7×102,respectively,compared with sialyl-Lewisx.CONCLUSION:Pregnancy-associated glycoproteins,hCG and glycodel in,are the powerful inhibitors for E-selectin-mediated endothelial cell adhesion.     

Effect of microwave ablation of liver cancer on cellular immunity in mice

AIM:To investigate the effect of microwave ablation of liver cancer on the cellular immunity in mice.METHODS:A C57BL/6J mouse model of liver cancer was established by subcutaneous injection of Hepa 1 - 6 cells.The tumors were subjected to micr owave ablation under the ablation condition of 45 ℃,50 ℃,55 ℃ or 60 ℃ fo r 180 s.The CD4＋ T cells,CD8＋ T cells and natural killer cells (NK) in p eripheral blood were detected by FACS.The cytotoxicity of splenic NK and splen ic cytotoxic T lymphocytes (CTL) activated by inactivated Hepa 1-6 cells was ass ayed by LDH method.RESULTS:The proportions of CD4＋ T cells,CD8＋ T cells and NK cells in peripheral blood in 50 ℃ and 55 ℃ group at 21 d after ablation we re significantly increased and that of NK cells in 60 ℃ group was significantly in creased.There was no significant difference between those in group 42 d after ablation and control.The cytotoxicities of splenic CTL and NK cells in 50 ℃ an d 55 ℃ groups at 21 d or 42 d after ablation were significantly increased,and they were much higher than those in 45 ℃ group at the same time.The cytotoxicities of splenic CTL in 50 ℃ and 55 ℃ groups at 21 d after ablation were much highe r than that in 60 ℃ group at the same time.CONCLUSION:Under a certain ablation temperature,microwave abla tion of liver cancer promotes the cellular immunity.     

The effect of mifepristone on natural killer subsets of peripheral blood and decidua in early pregnancy

AIM: To investigate the effect of mifepristone on natural killer(NK) subpopulations of peripheral blood and decidua in early pregnancy. METHODS: Flow cytometry was used to detect the expression of CD56 and CD16 on lymphocytes of decidua and peripheral blood in early pregnancy, mifepristone-treated pregnancy. RESULTS: The percentages of different natural killer subsets in peripheral blood between early pregnancy and mifepristone-treated pregnancy were almost identical. The serum levels of estradiol and progesterone in mifepristone-treated pregnancy were slightly higher than in early pregnancy. The percentage of decidual CD56⁺NK cell in early pregnancy was significantly higher than in mifepristone-treated pregnancy. The CD56⁺NK cells were predominant lymphocyte population of decidua in mifepristone-treated pregnancy, but in which the CD56⁺CD16⁺ and CD16⁺NK cells were major lymphocyte subpopulations of peripheral blood. CONCLUSION: Mifepristone acted principally on feto-maternal interface, it blocked the proliferation and differentiation of decidual CD56⁺NK cells and induced embryo immune rejection.     

Expression and effect of glucocorticoid receptor β on children with idiopathic nephrotic syndrome

AIM: To elucidate the significance of glucocorticoid(GC) receptor isoform β(GRβ)in children with idiopathic nephrotic syndrome(INS), and to evaluate the effect of sera from GC-resistant INS on the expression of GRβ. METHODS: The percentage of GRβ positive staining peripheral blood mononuclear cells(PBMC) and the quantity of nuclear protein of GRβ in PBMC were detected by immunocytochemistry and Western blotting assay, respectively. The effect of sera isolated from children with GC-resistant INS on GRβ expression was examined by cell culture in vitro. RESULTS: The number of GRβ positive staining PBMC and the quantity of nuclear protein of GRβ in children with GC-resistant INS were significantly higher than those in patients with GC-sensitive INS(P<0 01). The quantity of GRβ protein in nuclei of PBMC in children with GC-resistant INS was significantly higher than that in normal control and children with GC-sensitive INS. After 24 hours incubation with sera from children with GC-resistant INS, the percentage of GRβ positive PBMC and the quantity of nuclear GRβ protein in children with GC-sensitive INS and normal control increased significantly(P<0.05). CONCLUSION: Increased expression of GRβ is closely related with the GC resistance in children with INS, and high expression of GRβ is mediated, at least in part, by certain humoral factors in the serum.     

Activated NK cells against human hepatocellular carcinoma cell lines in vitro and in vivo

AIM: To evaluate the role of natural killer(NK) cells against a variety of human hepatocellular carcinoma (HCC) cell lines in vitro and in vivo, and to investigate the expression of MHC class I chain-related protein (MIC protein) in these HCC cell lines. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from 50 mL peripheral blood donored by a healthy volunteer and cultured in the NK cell kit followed by amplification and cytokine activation. The release of lactate dehydrogenase (LDH) was detected by lysis experiment. Animal experiments were performed following vaccination with HCC cell lines (BEL7402, HepG2 and SMMC7721) in BALB/c-nu/nu mice. The diameters of the tumors were measured and the growth curves of difference cell lines were delineated. Three weeks later, these mice were sacrificed and the weight of the transplanted tumors was also detected. Furthermore, the expression of MIC protein was determined. RESULTS: The obvious differences of lysis effects of NK cells on different cell lines were observed, in which the lysis effect of NK cells on K562 cells was the strongest, the effect on BEL7402 cells was in the middle and the effect on SMMC7721 cells was the weakest. In animal experiments, NK cells also showed obvious and different restraining effects on a variety of HCC cell line-transplanted tumors in nude mice. The tumor inhibitory rates of NK cells were 43.5% in BEL-7402 cell-transplanted tumor, 40.7% in HepG2 cell-transplanted tumor and 36.0% in SMMC-7721 cell-transplanted tumor. The protein expression of MIC was 48.7%, 32.8% and 0.9% in the 3 cell lines,respectively. CONCLUSION: The lysis effects of NK cells on a variety of human HCC cell lines are different. The expression of MIC in the tumor cells may play distinct role in evaluating these differences.     

HCMV-infected human THP-1 cells induce expression of HLA-G and its receptors

AIM: To investigate the differential expression of human leukocyte antigen-G (HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus (HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism.METHODS: THP-1 cells were infected with HCMV Towne strain. The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively. The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry. The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS: After infection of the THP-1 cells with HCMV, no obvious apoptosis in the cells was observed, and the viability of the cells was high. A significant up-regulation of HLA-G1, -G3, -G4 and -G5 at mRNA expression level 1 d after infection was found, while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected. The expression of HLA-G/ILT2/ILT4 was evidently up-regulated 1 d after infection. The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01). The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION: The differential expression of HLA-G isoforms and secretion of the receptors ILT2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection. This study provides experimental evidence for evaluating the immune mechanism of HCMV infection.     

Effect of pregnancy uterine microenvironment on the expression of NKG2A,NKG2D and their ligands in decidual NK cells

AIM：To investigate the expression of NKG2A,NKG2D and their ligands in pregnancy uterine micro-environment and to probe the function of NKG2A and NKG2D imbalance expression during the immunotolerance at the fetal-maternal boundary.METHODS：Decidual lymphocytes and peripheral lymphocytes were obtained from 30 women during 6-9 weeks of pregnancy who were undergoing selective termination.FACS technology was used to detect NK cells number and NKG2A,NKG2D expression.RT-PCR was used to investigate HLA-E and MICA mRNA in trophoblast tissue.RESULTS：Natural killer cells predominate,accounting for 70% of pregnancy endometrial lymphocytes.FACS results indicated that NKG2A was significantly increased in decidual NK cells as compared with that in peripheral NK cells,accounting for 97.86%±1.75% and 33.35%±10.92%.The difference between them in NKG2A expression was significant (P<0.05).Although the levels of NKG2D in decidual NK cells were similar to that in peripheral NK cells,accounting for 93.21%±4.52% and 97.80%±1.72%,respectively,the difference between them in NKG2A expression was also significant (P<0.05).At the same time,we found HLA-E mRNA in the trophoblast tissue.There was no proof yet on the expression of NKG2D ligand-MICA on trophoblast.CONCLUSIONS：Natural killer cells are the predominant lymphocytes during the first trimester of decidual tissue.These cells have a great different antigenic phenotype with peripheral natural killer cells.Decidual natural killer cells have high expression of NKG2A and trophoblast tissue expressed HLA-E.This may be an important factor contributing to the immunotolerance at the fetal - maternal boundary during pregnancy.     

Immune function of dendritic cells in patients with hepatocellular carcinoma

AIM: To investigate the immune function of dendritic cells (DCs) in patients with hepatocellular carcinoma(HCC). METHODS: The DCs were cultured from human peripheral blood mononuclear cells (PBMC) by using GM-CSF and IL-4 for 7 days. Surface molecules CD86, HLA-DR of DCs were detected by flow cytometry. IL-12 production by DCs and IFN-γ production by T cells was measured with ELISA and ELISPOT, respectively. Allogenic mixed lymphocyte reaction was detected by MTT assay. RESULTS: CD86 expression in DCs in HCC patients were markedly lower than that in health control (91.7％ vs 83.5%, P<0.05). IL-12 production by DCs had no significant difference between the HCC patients and the health control ［(324.6±171.0)ng/L vs (436.5±142.7)ng/L, P>0.05］. However, after stimulated DCs with LPS, IL-12 production in HCC patients was significantly lower than that in health control ［(478.6±142.7)ng/L vs (630.0±151.9)ng/L, P<0.05］. IFN-γ production by T cells and T cell proliferation index (PI) in the HCC patients were all significantly lower than those in health control ［(IFN-γ: 133.4±51.2)103U/L vs (183.0±60.2)103U/L, P<0.05; PI: 2.3±0.7 vs 3.5±0.8, P<0.01］. CTL killing activity assay indicated that DC-induced CTL killing activity in HCC group was significantly lower than that in health group when the E/T ratio was 10∶1 and 20∶1, respectively. CONCLUSION: The immune function of DCs and T cells in the patients with HCC are significantly decreased. The CTL killing activity of HCC group is also markedly decreased.     

The expression of hHR21sp gene in the peripheral blood cells and bone marrow cells of patients with Fanconi anemia

AIM:To detect the expression of hHR21^spgene in normal human peripheral blood cells and in hemapoietic cell from fanconi anemia(FA) patients bone marrow after exposure to UV and gamma-irradiation.METHODS:RNA of FA hemapoietic cell and normal peripheral blood cells were harvested after UV and gamma irradiation. hHR21^sp gene expression was analyzed using RT-PCR, southern blot hybridization, phosphoimmage autoradiogram.RESULTS:The expression of hHR21^sp gene in PBL varied with the time of UV gamma-irradiation and increased obviously six hours after 80 J/M² UV-or 5 Gy gamma-irradiation. However, the expression was down-regulated in FA hemapoietic cell from FA patients indicating that hHR21^sp expression is something related to the ability of cells to repair DNA double strand break.CONCLUSION:The expression of hHR21^sp down-regulated in the FA syndrome and this might be involved in the pathogenesis of the FA.     

Value of single nucleotide polymorphism array in investigating origin and pathogenic mechanisms of uniparental disomy

AIM：To assess the value of the genome-wide human single nucleotide polymorphism (SNP) array for the investigation of the origin and pathogenic mechanisms and the genetic counseling of uniparental disomy (UPD). METHODS：Genetic analysis with the genome-wide human SNP array was carried out on the fetal cells in amniotic fluid and the peripheral blood cells from 124 pregnant women with high risk of Down’s syndrome, whose G-banded chromosome karyotype analysis was done using the fetal cells in amniotic fluid. The peripheral blood cells from the fetuses fathers were also taken for SNP array analysis. RESULTS：Two cases of segmental UPD 16 were found from the SNP array analysis of the fetal cells in amniotic fluid. The regions of isodisomy in one case were located in 16p12.2~13.3 and 16q24.1~24.3, and the region of the other was located in 16q21~24.3. Both cases of UPD were maternal upon the genetic linkage analysis of the peripheral blood cells of the parents. CONCLUSION：The genes that induce the fetal growth restriction are probably located at the ends of long arm and short arm of chromosome 16. SNP array can identify the parental origins and the pathogenic mechanisms of UPD, which provides the assistance for genetic counseling.