OBJECTIVETo study the relationship between hepatitis B immunoglobulin (HBIG) injection before delivery and hepatitis B virus (HBV) S gene mutation.

METHODS18 neonates infected with HBV in uterus and their mothers were divided to a) HBIG group (8) in which their mothers received HBIG injection before delivery and b) control group (10) in which their mothers never received any treatment HBV DNA fragments were amplified by nest-PCR from sera of these neonates and their mothers. S gene region of these HBV DNA fragments were directly sequenced and data on mutations was analyzed.

RESULTSThere was no significant difference on nucleotide and amino acid changes in the S gene between the HBIG group and the control group. The majority HBV strains of newborn (17/18) were identical to their mother's dominant strains before delivery, including four mutation HBV strains. Among 18 newborns with HBV intrauterine infection, 12 were infected by B type (adw2), and 6 by C type (adrq+).

CONCLUSIONMothers who were asymptomatic HBsAg carrier and received injections ofHBIG before delivery would not be influenced by HBV S gene mutation. HBV intrauterine transmission with or without gene mutation might occur in the third-trimester of pregnancy. Gene mutation of HBV was not the main factor in intrauterine transmission of HBV.

Female ; Genes, Viral ; Hepatitis B ; prevention & control ; transmission ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Immunoglobulins ; administration & dosage ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Mutation ; Pregnancy

OBJECTIVETo reassess the diagnostic value of 24 hour urinary copper excretion in children with Wilson disease (WD).

METHODSFrom July 2005 to June 2007, inpatients over three years old in a pediatric liver center were assigned into WD and non-WD group.

RESULTS94 patients, including 26 cases in WD and 68 in non-WD group, were enrolled in this study. The median of 24 h urinary copper excretion was 98.5 microg in WD group and 25.8 microg in the non-WD group (Z = -6.111, P equal to 0.000). The area under receiver operator curve (ROC) was 0.909 (95% CI: 0.839-0.979, P equal to 0.000). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 84.6%, 91.2%, 89.4%, 78.6% and 93.9% respectively using 52.0 ug as a cutoff value, and 50.0%, 97.1%, 84.0%, 86.7% and 83.5% using 100 microg as a cutoff value. The goodness of fitness of 52 microg criteria was significantly higher than 100 microg criteria (kappacoefficient 0.760, 0.541 respectively, P equal to 0.000).

CONCLUSIONComparing to 100, 52 microg of 24 h urinary copper excretion as a cutoff value significantly improves the sensitivity and accuracy for diagnosing WD in children.

Adolescent ; Age Factors ; Ceruloplasmin ; Child ; Child, Preschool ; Copper ; urine ; Female ; Hepatitis ; diagnosis ; pathology ; urine ; Hepatitis A ; diagnosis ; pathology ; urine ; Hepatolenticular Degeneration ; diagnosis ; pathology ; urine ; Humans ; Liver ; pathology ; Male ; Penicillamine ; Predictive Value of Tests ; ROC Curve ; Sensitivity and Specificity ; Time Factors

To study the relationship between tau hyperphosphorylation and the function of glutamate transporter okadaic acid (OA), a protein phosphatase inhibitor, 20 ng in a 0.5 microl volume, was injected into the frontal cortex of rat brain and immunostaining was used to observe the phosphorylation of tau protein and the expression of excitatory amino acid transporter 1 (EAAT1) in the brain following the injection. The results showed that (1) the neurons in the center of the injection region displayed cytoplasmic shrinkage, swelling, nuclear pyknosis, and dislocation at the early stage, and necrosis appeared 3 d after the injection. However, most neurons in the peri-injected areas showed normal morphological characters with immuno positive reaction for AT8, a tau phosphorylated marker; (2) morphological analysis showed that tau hyperphosphorylation caused by OA treatment was mainly observed in the axons and dendrites of neuronal cells at 6 h in the cell body at 1 d, which brought about dystrophic neurites and neurofibrillary tangle (NFT)-like pathological changes; (3) the induction of glutamate transporter EAAT1 was observed in the involved areas corresponding to that with AT8 immunopositive staining, and the number of EAAT1-positive staining cells markedly increased at 12 h (P<0.01), peaked at 1 d (P<0.001), then decreased at 3 d following the injection. Combined with a confocal laser scanning microscopic analysis, double fluorescent immunostaining showed that EAAT1 positive staining appeared in neurons as well as astrocytes in the peri-injected areas of the frontal cortex. These results demonstrate that OA increases glutamate transporter EAAT1 expression in neurons while it induces tau hyperphosphorylation. However, the mechanism and significance of the induction of glutamate transporter EAAT1 expression remain to be further elucidated.
Animals ; Astrocytes ; drug effects ; metabolism ; Axons ; drug effects ; metabolism ; Brain ; cytology ; Dendrites ; drug effects ; metabolism ; Excitatory Amino Acid Transporter 1 ; metabolism ; Neurofibrillary Tangles ; pathology ; Neurons ; drug effects ; metabolism ; Okadaic Acid ; pharmacology ; Phosphorylation ; Rats ; tau Proteins ; metabolism

OBJECTIVETo explore the efficacy of intraperitoneally injected epirubicin (EPI)-loaded poly (d, l)-lactic acid (PLA) microspheres (MS) alone or combined with free epirubicin (FEPI) in treating hepatocellular carcinoma (HCC) in mice.

METHODSMice that were transplanted with H22 ascites HCC were randomized into seven groups, which were intraperitoneally injected with blank microspheres, normal saline, three different doses of microspheres (9, 18, and 36 mg/kg EPI) , FEPI (9 mg/kg) , and the combination (microspheres with EPI 4.5 mg/kg + FEPI 4.5 mg/kg). The survival time of all animals was recorded. The rates of increase in life span of all the treatment groups were calculated.

RESULTSEPI-PLA-MS significantly prolonged the survival time of HCC mice in a dose-dependent manner, with a maximal tolerated dose (MTD) of 18 - 36 mg/kg. The combination group had the highest average survival time, median survival time, and rate of increase in life span, which were (40.0 +/- 16.9) days, 33.5 days, and 222.58%, respectively.

CONCLUSIONEPI-PLA-MS combined with FEPI is highly effective in treating HCC in mice when intraperitoneally injected.

Animals ; Delayed-Action Preparations ; Epirubicin ; administration & dosage ; therapeutic use ; Infusions, Parenteral ; Lactic Acid ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Microspheres ; Polyesters ; Polymers

AIM:To analyze zebrafish embryos and to identify erythropoietin (Epo) as an active renal anti-apoptotic factor in an Epo receptor (EpoR)-dependent manner. METHODS:For transient knockdown of Epo and EpoR in renal Tg (wt1b:EGFP) zebrafish reporter lines, the morpholino antisense oligonucleotide technique was used. Morphant zebrafish embryos were phenotypically analyzed using fluorescence microscopy. Apoptosis was determined by TUNEL assay and Annexin V staining. Western blot was used to identify Akt phosphorylation in Epo and EpoR knockdown zebrafish. RE-SULTS:Epo and EpoR zebrafish morphants exhibited pathophysiological changes within the pronephros, adversely affecting pronephric structure. Zebrafish embryos upon silencing of Epoa and EpoR showed a significant increase in the apoptosis within the zebrafish pronephros, consequently leading to renal pathophysiological effects. Decreased p-Akt was identified in Epo and EpoR knockdown zebrafish embryos. CONCLUSION:Epo is an essential regulator of renal development and function by interacting with its receptor EpoR and thereby repressing apoptosis, mechanistically by promoting Akt phospho-rylation.

Objective To establish simple,stable and reliable rat models of oxygen glucose deprivation/reoxgenation(OGD/R)in adult neural stem cells(NSCs)in vitro.Methods The NSCs from adult Fisher344 rats were cultured in serum-free medium and identified using nestin and DAPI immunofluorescent double staining.These cells were washed with a Earle′s balanced salt solution without glucose for 2 times,then,incubated for different periods(2,4,6,8 and 10 h)in a trigas incubator with an atmosphere of 1％ O2,5％CO2 and 94％ N2,98％ humidity at 37 ° C.And then,these cells were removed from the anaerobic incubator,washed,and added DEME/F12 containing bFGF supplement.A normoxic-normoglycemic control group was employed.Morphological assessment of NSCs was performed by light microscopy after re-oxgenation for 24 h; CCK-8 colorimetric method was used to determine the survival and proliferation of NSCs,and flow cytometry was employed to detect the apoptosis of NSCs.Results After the setting of oxygen glucose deprivation for 2 h,the OD value and the survival rate in the OGD cells were increased as compared with those in control group without significant difference(P＞0.05).While the morphological damage of NSCs aggravated gradually and the OD value decreased in OGD cells following the prolongation of times; under the setting of oxygen glucose deprivation for 6 h,the OD value in OGD cells obviously decreased as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the survival rate obviously decreased and the apoptosis rate significantly increased in OGD cells as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the apoptosis rate of NSCs excessed to 50％.Conclusion By means oftrigas incubator,simple,stable and reliable models of OGD/R in NSCs in vitro can be successfully established.

OBJECTIVETo study the effectiveness of treating hepatocellular carcinoma (HCC) in mice with locally administered epirubicin-loaded poly( D, L) - lactic acid microspheres (EPI-PLA-MS ).

METHODSEPI-PLA-MS was prepared with double emulsion solvent evaporation technique. Five groups of mice (n = 8 in each group) were intraperitoneally injected with five different doses of free epirubicin (FEPI), and the maximum tolerated dose (MTD) was calculated. Then 15 mice with transplanted subcutaneous H22 HCC were divided into three groups (n = 5), which were respectively intratumorally injected with normal saline (NS), blank microspheres, and EPI-PLA-MS (with 9 mg/kg of EPI). After two weeks the tumors were excised and weighed. Another 15 mice with transplanted H22 ascites HCC were divided into three groups (n = 5), which were intraperitonealy injected with the same drugs, and the increased life span were registered exactly.

RESULTSThe MTD of intraperitoneally injected FEPI was 9 mg/kg. The tumour-inhibiting rates was 40.35% and 36.09% when EPI-PLA-MS were administered by intratumoral injection to the mice with subcutaneous H22 HCC. It significantly prolonged the survival time of mice with H22 ascites HCC and the increased life span by 153.49% and 142.22% when EPI-PLA-MS were intraperitoneally administered.

CONCLUSIONEPI-PLA-MS is a new sustained-release preparation with high-efficacy and low-toxicity in treating HCC and has shown promising prospects when administered locally.

Animals ; Antibiotics, Antineoplastic ; administration & dosage ; Delayed-Action Preparations ; Drug Carriers ; Epirubicin ; administration & dosage ; Injections, Intraperitoneal ; Lactic Acid ; pharmacology ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Mice, Inbred Strains ; Microspheres ; Polyesters ; Polymers ; pharmacology

OBJECTIVETo explore the possible relationship between cytokines (TNF-alpha, IFN-gamma, IL-4 and IL-10), which were expressed abnormal quantity in the peripheral blood to intrauterine HBV infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to HBV intrauterine infection.

METHODSA cross sectional study on molecular epidemiology was carried out. The subjects were selected from outpatients of the hepatitis B vaccine special clinics of our hospital. According to intrant criteria, children under high risk of HBV intrauterine infection were divided into immuno-failure group (group I) and immuno-effective group (group II) while children without high risk were included in the control group. Four gene SNP sites of TNF-alpha-238, IFN-gamma + 874, IL-4-590 and IL-10-1082 region were determined by real-time quantitative fluorescent PCR.

RESULTSSignificant differences of TNF-alpha-238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) as well as between group I and control group (chi(2) = 9.513, P < 0.05). No evident difference of TNF-alpha-238 A was found between group II and control group (chi(2) = 0.047, P > 0.05). Significant differences of IFN-gamma + 874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the controls (chi(2) = 5.199, P < 0.05) but no significant difference was found between group II and control group (chi(2) = 0.602, P > 0.05). Significant differences of IL-4-590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), group I and control group (chi(2) = 0.584, P > 0.05), or between group II and control group (chi(2) = 0.004, P > 0.05) respectively. Significant differences of IL-10-1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but not found between group I and control group (chi(2) = 1.759, P > 0.05).

CONCLUSIONThis study suggested the possibility that TNF-alpha-238 A allele and IFN-gamma + 874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4-590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10-1082 G allele seemed to be associated with preventive efficacy to HBV intrauterine infection.

Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Hepatitis B ; transmission ; Humans ; Infant ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Polymorphism, Genetic ; Pregnancy ; Pregnancy Complications, Infectious ; Retrospective Studies ; Risk Factors ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo study the cytotoxic effect of allogenetic natural killer (NK) cells in vitro on human CD34+ acute myelogenous leukemia cells.

METHODSCD34 expression on acute myelogenous leukemia KG1a cells was detected by flow cytometry. KG1a cells were co-cultured at different effector-to-target (E:T) ratios with NK cells isolated from 5 healthy individuals using magnetic cell sorting. Lactate dehydrogenase (LDH) release assay was employed to examine the cytolysis of KG1a cells in the co-culture, and the inhibition rate of the KG1a cell colony formation in methylcellulose was determined with K562 cells sensitive to NK cells as the control.

RESULTSA expression rate as much as (98.0-/+1.1)% was detected for CD34 antigen on KG1a cells, and the isolated NK cells (CD3(-)CD16+CD56+ cells) had a purity of (93.2-/+3.7)% after magnetic cell sorting. Allogenetic NK cells exhibited obvious cytotoxicity and colony inhibition in vitro against KG1a cells at different E:T ratios, and the effects were significantly enhanced as the E:T ratios increased (P<0.05). At the same E:T ratio, the cytotoxicity and colony inhibition rate of allogenetic NK cells against KG1a cells was lower than those against K562 cells (P<0.05).

CONCLUSIONAllogenetic NK cells exhibit obvious cytotoxicity and colony formation against CD34+ acute myelogenous leukemia cells.

Antigens, CD34 ; immunology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Flow Cytometry ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology

BACKGROUNDThe influences of genomic background are confirmed in more diseases. Immunologic tolerance after intrauterine infection of hepatitis B virus is considered to occur in T cells. Cytokines work effectively in eliminating virus by immune system after hepatitis B virus infection. To explore the relationship between cytokines (tumor necrosis factor-alpha, interferon-gamma, interleukin-4 and interleukin-10), which expressed abnormal quantity in the peripheral blood to intrauterine hepatitis B virus infectious children, gene single nucleotide polymorphism (SNP) and susceptibility to hepatitis B virus intrauterine infection.

METHODSThis is a cross sectional study of molecular clinical epidemiology. The subjects in this study were selected from outpatients of hepatitis B vaccine follow-up special clinics of our hospital in the period. According to intrant criteria, the high risk children of hepatitis B virus (HBV) intrauterine infection were divided into immune failure group (group I); and immune effective group (group II) and non high risk children belonged to the control group. Four gene SNP sites of TNF-alpha -238, IFN-gamma +874, IL-4 -590 and IL-10 -1082 were determined by real-time quantitative fluorescent polymerase chain reaction (PCR).

RESULTSThe significant differences of TNF-alpha -238 A allele frequency were found between group I and group II (chi(2) = 6.797, P < 0.05) and between group I and the control group (chi(2) = 9.513, P < 0.05). No evident differences of TNF-alpha -238 A were found between group II and control group (chi(2) = 0.047, P > 0.05); the significant differences of IFN-gamma +874 A allele frequency were found between group I and group II (chi(2) = 7.238, P < 0.05), and between group I and the control group (chi(2) = 5.199, P < 0.05). No evident differences were found between group II and the control group (chi(2) = 0.602, P > 0.05); the significant differences of IL-4 -590 C/T allele frequency were not found between group I and group II (chi(2) = 0.632, P > 0.05), also group I and the control group (chi(2) = 0.584, P > 0.05), and the group II and the control group (chi(2) = 0.004, P > 0.05) respectively; The significant differences of IL-10 -1082 G allele frequency were found between group II and group I (chi(2) = 10.359, P < 0.001), and between group II and the controls (chi(2) = 35.418, P < 0.001), but the significant differences were not found between group I and the control group (chi(2) = 1.759, P > 0.05).

CONCLUSIONSThis study suggested the possibility that the TNF-alpha -238 A allele and IFN-gamma +874 A allele were associated with HBV intrauterine infection. There was no evident relationship between IL-4 -590 C/T allele SNP and susceptibility to HBV intrauterine infection, but the IL-10 -1082 G allele was associated with preventive efficacy to HBV intrauterine infection.

Cross-Sectional Studies ; Cytokines ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Hepatitis B ; genetics ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Interleukin-4 ; genetics ; Male ; Pregnancy ; Tumor Necrosis Factor-alpha ; genetics