Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

OBJECTIVETo study the effect of large-volume liposuction on adipokines, inflammatory markers and insulin resistance and its significance.

METHODSThe levels of serum adiponectin, resistin, IL-6, TNF-alpha, CRP and insulin sensitivity were measured pre- and 3 months postoperatively in 16 consecutive patients undergoing liposuction.

RESULTSThe adiponectin postoperative [(5.56 +/- 1.03) ng/ml] was significantly higher than the preoperative [(6.54 +/- 1.27) ng/ml] (P<0.05); the resistin postoperative [(6.30 +/- 3.25) pg/ml] is significantly lower than the preoperative [(5.18 +/- 2.28) pg/ml] (P<0.05); the insulin sensitivity postoperative (-3.78 +/- 0.39) significantly increased compared to the preoperative (-3.48 +/- 0.32, P<0.05); the TNF-alpha postoperative [(0.19 +/- 0.02) pg/ml] significantly increased compared to the preoperative [(0.23 +/- 0.03) pg/ml, P<0.01]; the postoperative IL-6 [(0.27 +/- 0.04) pg/ml] was similar to the preoperative [(0.31 +/- 0.06) pg/ml, P >0.05]; the postoperative CRP [(8.03 +/- 6.01) mg/L] was not significantly different with the preoperative [(7.28 +/- 6.21) mg/L, P>0.05].

CONCLUSIONSLarge-volume liposuction has good effect on insulin resistance and adipokines.

Adipokines ; blood ; Adiponectin ; blood ; Adult ; Female ; Humans ; Insulin Resistance ; Interleukin-6 ; blood ; Lipectomy ; Lipid Metabolism ; Middle Aged ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVEDendritic cell vaccines are one of the important active immunotherapies for neoplasms. The aim of this study was to observe the killing effect of specific cytotoxic T lymphocytes (CTL) on liver carcinoma HepG2 and SMMC-7721 cells in vitro. The CTL was induced by human peripheral blood mononuclear cells-originated dendritic cells (DC) transfected by recombinant adeno-associated virus (rAAV) with hAFP gene fragment (137-145).

METHODSImmature DCs were generated from peripheral blood mononuclear cells of healthy volunteers and then transfected by rAAV with AFP gene fragment. The CTL was thereafter induced. The activities of DC and CTL were measured and the killing effect of the CTL on HepG2 cells was detected using M1Tr assay.

RESULTSThe mature DC, transfected or not, highly expressed CD40, CD86 and IL-12. IFN-gamma was highly expressed in the CTL. The DC-induced CTL could effectively recognize and destroy the HepG2 and SMMC-7721 cells.

CONCLUSIONDC transfected by rAAV can stimulate the proliferation and differentiation of lymphocytes and also induce the proliferation of CTL, and their own phenotypes are not significantly altered. The DC vaccine can be effectively used as an adjuvant immunotherapy for patients with hepatocellular carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Dependovirus ; genetics ; Hep G2 Cells ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Liver Neoplasms ; pathology ; Peptide Fragments ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; pathology ; Transfection ; alpha-Fetoproteins ; genetics

OBJECTIVETo study the effect of liposuction on insulin resistance and lipid metabolism.

METHODSThe levels of serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and insulin sensitivity were measured pre-and 2-4 months postoperatively in 20 consecutive patients undergoing liposuction.

RESULTSCompared with preoperative, the insulin sensitivity increased significantly, the levels of TC and LDL-C decreased after the liposuction procedure.

CONCLUSIONSLiposuction may improve the insulin resistance and lipid metabolism.

Adult ; Blood Glucose ; metabolism ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Humans ; Insulin Resistance ; Lipectomy ; Lipid Metabolism ; Middle Aged ; Triglycerides ; blood ; Young Adult

OBJECTIVETo explore the effect of retinoblastoma binding protein 2 (RBP-2), a histone H3K4 demethylase, on osteogenic differentiation of human adipose-derived stromal cell (hASC).

METHODSAccording to the GenBank sequence information of RBP-2, four different small interfering RNAs (siRNA) targeting RBP-2 gene were designed and the corresponding short hairpin RNAs (shRNA) were cloned into pLL 3.7 lentivirus RNA interference vector. The lentivirus with RBP-2-siRNA was packaged in 293T cells. The effective sequence was examined and selected by Western blotting and real-time PCR. The lentiviruses with efficient knockdown effects were used to infect hASC. On the 14th day after osteogenic differentiation, alkaline phosphatase (ALP) activities of hASC were quantitatively tested and at the same time, ALP staining and alizarin red staining were performed to assess the difference of osteogenic differentiation between the knockdown group and the control group.

RESULTSThe recombinant lentivirus siRNA targeting RBP-2 was successfully constructed and the expression of RBP-2 mRNA and protein were dramatically suppressed by infection with RBP-2-siRNA lentivirus. On the 14th day after osteogenic induction, ALP activity of hASC in the knockdown group [(299.2 ± 22.7), (224.3 ± 17.7) U/g] was much stronger than that in the control group [(129.9 ± 12.9) U/g, P < 0.05] and the same result was achieved for the ALP staining and alizarin red staining.

CONCLUSIONSThe constructed RBP-2-siRNA lentivirus could markedly decrease the expression of RBP-2 and promote osteogenic differentiation of hASC. It indicated that RBP-2 can repress the osteogenic differentiation of hASC.

Adipose Tissue ; cytology ; Adult ; Alkaline Phosphatase ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Cells, Cultured ; Female ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Lentivirus ; Osteogenesis ; Osteosarcoma ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Retinoblastoma-Binding Protein 2 ; genetics ; metabolism ; Stromal Cells ; cytology ; metabolism

Humans ; Inflammation ; etiology ; Sepsis ; complications ; Signal Transduction

OBJECTIVETo evaluate the therapeutic effects of bicyclol combined with ganciclocir on infantile cytomegalovirus hepatitis.

METHODSSeventy infants with cytomegalovirus hepatitis were randomized into treatment group (n=35) and control group (n=35) for a 2-week-long treatment with ganciclocir (5 mg/kg) with and without oral bicyclol (3 mg/kg, twice daily), respectively.

RESULTSIn both groups, significant changes occurred in the levels of alanine aminotransferase, alkaline phosphatase, serum total bilirubin, serum total bile acid, and glutamyl transpeptidase after the 2-week treatment (P<0.01); these parameters differed significantly between the two groups after the treatment (P<0.01). Compared with those in the control group, the infants in the treatment group showed significantly better responses to the treatment (P<0.05) with a significantly higher rate of serum anti CMV IgM negativity (P<0.05).

CONCLUSIONSBicyclol combined with ganciclocir can reduce glutamic pyruvic transaminase, alkaline phosphatase and serum total bilirubin, and decrease bile acid levels to lessen liver cell damage and promote the recovery of liver cells.

Alanine Transaminase ; metabolism ; Alkaline Phosphatase ; metabolism ; Antiviral Agents ; therapeutic use ; Bilirubin ; blood ; Biphenyl Compounds ; therapeutic use ; Cytomegalovirus ; Cytomegalovirus Infections ; drug therapy ; Drug Therapy, Combination ; Ganciclovir ; therapeutic use ; Hepatitis ; drug therapy ; virology ; Humans ; Infant ; Liver Function Tests

P13-OMP (29.1). P13-OMP and OMP68 group challenged with P13 and P11 can be efectivly protected; P13-WCB group challenged with P13 and P11 can not be efectivly protected; the control group were died out. The P13-OMP and OMP68 of Bordetella bronchiseptica has good immunogenicity and protection, so the results of this study lay good theoretical foundation for OMP subunit vaccine.

OBJECTIVETo explore the risk factors of acute lymphoblastic leukemia (ALL) recurrence in adult patients and establish a prognosis index (PI) calculation model in order to improve the prevention strategy of ALL in adults.

METHODS104 adult ALL patients from Blood Diseases Hospital & Chinese Academy of Medical Sciences between August 2008 and November 2011 were enrolled. COX proportional hazards regression stratified by Dummy variable was used to set up the prediction model; Kaplan-Meier method and Log-rank test were used to estimate and compare the survival. After calculated individual PI value, patients' expected survival should be estimated by groups.

RESULTSThe overall median survival of adult ALL patients was 22.00 months (95% CI 17.00-27.00). COX regression analysis showed that chemotherapy group patients had a higher risk of recurrence than of ASCT group while setting treatment as the dummy variable (RR=2.052, 95%CI 0.877-4.799, P=0.007). Stratified Analysis showed that the risk factors of B-ALL recurrence in adult patients included HGB <100 g/L (RR=0.186, 95% CI 0.068-0.512, P=0.001), CNSL (RR=7.767,95% CI 2.951- 20.433, P=0.001), number of consolidation chemotherapy<3 (RR=0.445, 95% CI 0.211-0.940, P=0.034) and Ph chromosome positive (RR=2.771, 95% CI 1.353-5.674, P=0.005). Grouped by the PI value, the expected survival of each individual patient could be estimated as PI=0.58 base.

CONCLUSIONHGB, CNSL, number of consolidation chemotherapy and Ph chromosome were independent risk factors of B-ALL recurrence in adult patients. PI value could predict the survival of adult ALL patients and provide reference for individual therapy and prognostic evaluation.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; epidemiology ; pathology ; Prognosis ; Recurrence ; Risk Assessment ; Risk Factors ; Young Adult

This study was to establish the episomal vector reprogramming method to reprogram iPSC from human cord blood (CB) CD34(+) cells. The non-integrating plasmids of pEB-C5 and pEB-Tg were transfected into short-term cultured CB CD34(+) cells by using the nucleofector, so as to demonstrate efficient reprogramming of CB CD34(+) cells. Within 14 days of one-time transfection by two plasmids together, up to 200 iPSC-like colonies per 2 million transfected CB CD34(+) cells were generated. The results showed that the pluripotency of iPSC-derived CB CD34(+) cells was similar to that of hESC and the karyotypes of iPSC were normal. In addition, no vector integration was found in iPSC of 9th and 10th passages. Furthermore, hiPSC formed teratoma with three embryonic germ layers. It is concluded that the integration-free method to generate human iPSC from CB CD34(+) cells is reliable and can provide new ways for both research and future clinical applications.
Animals ; Antigens, CD34 ; immunology ; Cell Culture Techniques ; Cells, Cultured ; Cellular Reprogramming ; Fetal Blood ; cytology ; immunology ; Fibroblasts ; cytology ; Humans ; Induced Pluripotent Stem Cells ; cytology ; Mice ; Plasmids