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 explore the susceptibility of children to develop intrauterine hepatitis B virus (HBV) infection through studying the association between interferon gamma (IFN-gamma) + 874 single nucleotide polymorphism (SNP) and intrauterine HBV infection.

METHODSThe subjects were selected from outpatients who were in our hepatitis B (HB) vaccine following-up clinics. The subjects whose mothers were HBV carriers were inoculated with HB vaccine or HB vaccine and hepatitis B immunoglobulin (HBIg). Intrauterine HBV infection was defined as peripheral blood HBsAg and/or HBV-DNA positive at birth and lasting for six months (group I). Normal immune children were defined as peripheral blood negative for HBV marker since birth and afterwards HBsAb titers were above protective level (group II). The subjects were composed of the following two groups. Group I consisted of 46 children with intrauterine HBV infection. Group II was composed of 73 normal children. A Taqman fluorescence polymerase chain reaction for the IFN-gamma + 874 SNP was performed for both groups.

RESULTSIFN-gamma + 874 SNP was tested successfully for every subject. Frequencies of AA, AT and TT genotype were 67.4%, 19.6% and 13.0% in the intrauterine HBV infection group, and 45.2%, 30.1% and 24.7% in the normal immune children group. A significant difference was found in the frequency distribution of IFN-gamma + 874 genotype between the two groups (chi(2) = 5.102, P = 0.02389). In the intrauterine HBV infection group the AA genotype was more common than in normal immune group.

CONCLUSIONThere is an association between IFN-gamma + 874 SNP and intrauterine HBV infection. This study suggested the possibility that IFN-gamma + 874 SNP might be important in determining an individual's susceptibility to development of intrauterine HBV infection.

Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Hepatitis B ; genetics ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Interferon-gamma ; genetics ; Male ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide

OBJECTIVETo study the possible relationship between tumor necrosis factor (TNF)-alpha-238G/A gene polymorphism and the susceptibility to intrauterine HBV infection.

METHODSTwo hundred and fifty-six children, including 130 infants born to HBsAg positive mothers were divided into two groups: forty-five children with intrauterine HBV infection (group I) and 85 children without intrauterine HBV infection (group II), with a control group of 126. TNF-alpha-238G/A gene polymorphism was examined in all 256 children, by means of real-time quantitative fluorescent PCR.

RESULTSA significant difference of TNF-alpha-238A allele frequency was found between group I and group II (x2=6.797, P=0.009), and between group I and the controls group (x2=0.047, P=0.002), but there was no significant difference between group II and the control groups (x2=0.047, p=0.828).

CONCLUSIONThis study found that genetic polymorphism of tumor necrosis factor-a was associated with intrauterine HBV infection

Adult ; Child ; Disease Susceptibility ; Female ; Hepatitis B ; transmission ; Humans ; Infant ; Infectious Disease Transmission, Vertical ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; Tumor Necrosis Factors ; genetics

OBJECTIVETo evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms.

METHODSSixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-β1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing.

RESULTSIntraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-β1 and collagen I expressions in the ischemic tissues.

CONCLUSIONSIntraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-β signaling pathway.

Animals ; Cecum ; injuries ; pathology ; Collagen Type I ; metabolism ; Fibrinolysis ; Injections, Intraperitoneal ; Peritoneum ; injuries ; pathology ; Phenanthrenes ; pharmacology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Postoperative Complications ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Adhesions ; prevention & control ; Tissue Plasminogen Activator ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing

OBJECTIVETo evaluate the clinical effect of implanted cardioverter-defibrillators (ICD) for primary prevention of sudden cardiac death.

METHODSAccording to ACC/AHA Guideline of ICD implantation (2005), 35 patients successfully received ICD/CRT-D implantation for primary prevention of sudden cardiac death in our hospital from January 2006 to December 2009. All the patients were followed up for a mean of 2 years.

RESULTSDuring the follow up, 11 (31.43%) patients experienced ventricular arrhythmic episodes, for which 16 defibrillation therapies and 75 anti-tachycardia pacing (ATP) therapies were delivered without mistaken shock or death. The incidence rate of NVM was 100%, that of PVT was 66.67%, Brugada syndrome 50%, HCM 25% and DCM 16.67%. Of these episodes, the incidence of VF episodes among PVC patients was 87.5% (14 beats), ventricular tachycardia PVC was 82.28% (65 times), 5 beats in NVM patients, 4 beats in HCM and Brugada syndrome patients, and 1 beat in DCM patients. No ICD implantation-related complication was detected, and no ventricular tachycardia induced syncope occurred in these cases. All patients showed improved quality of life after the implantation.

CONCLUSIONICD implantation can prevent malignant ventricular arrhythmia episodes, especially for PVT, NVM and Brugada syndrome in high risk SCD patients, demonstrating the value of implantation of ICD as a primary prevention in high-risk SCD patients.

Adult ; Aged ; Death, Sudden, Cardiac ; epidemiology ; prevention & control ; Defibrillators, Implantable ; Female ; Humans ; Male ; Middle Aged ; Primary Prevention

OBJECTIVETo study the clinicopathologic features, immunophenotype and gene rearrangement of primary cutaneous diffuse large B-cell lymphoma, leg type (PCLBCL).

METHODSSeven cases of PCLBCL were enrolled into the study. Clinicopathologic analysis, immunohistochemical staining and gene rearrangement for IgH and Igκ were undertaken in the study.

RESULTSAll the seven cases were male, and the median age was 72 years. Patients usually presented with multiple purple tumors, nodules, papules and infiltrative plaques. Two patients had a history of leg injury before onset, and one had mosquito bites. Histologically, the tumor involved the dermis and subcutis with dense and diffuse infiltrative pattern composing of centroblasts and/or immunoblasts. Immunohistochemical staining showed that seven cases (7/7) expressed CD20, six (6/6) expressed bcl-2, four (4/4) expressed MUM-1, four (4/5) expressed CD79a, four (4/5) expressed PAX-5 and four (4/6) expressed bcl-6, respectively. All cases did not express CD3ε, CD45RO, CD10 and CD30. IgH gene rearranged bands were detected in three (3/6) cases and Igκ was detected in one (1/5) case. Six of the seven cases died and the remaining patient, who was 44-year-old, was alive after 22 months of follow-up.

CONCLUSIONSPCLBCL is rare, predominantly affects elderly male patients. PCLBCL has poor prognosis and high mortality, but younger patients seem to have better prognosis. Some cases had a history of trauma or mosquito bites. The relationship between the history and the onset of PCLBCL needs further evaluation.

Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; analysis ; Culicidae ; Gene Rearrangement ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Immunophenotyping ; Insect Bites and Stings ; complications ; Leg ; Leg Injuries ; complications ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Skin Neoplasms ; genetics ; pathology

OBJECTIVETo investigate the gene defect in a hereditary coagulation factor V (FV) deficiency family.

METHODSThe plasma FV actigen was measured by one-stage clotting assay. The FV antigen was assayed by Biotin-Avidin enzyme linked immunosorbent assay (BA-ELISA). The full length of exon 1 to exon 25 and the 5' untranslated sequence of FV genomic DNA were analyzed by polymerase chain reaction (PCR) and direct sequencing of the amplified fragments, meanwhile the defect was identified by T/A cloning sequencing.

RESULTSThe plasma coagulant activity and amount of FV of the proband were marked deficient (1% and 1.54%, respectively). DNA sequence analysis for the proband revealed a causative mutation in a heterozygous status. It was one base pair deletion in exon 4 at nucleotide 675 inherited from her mother.

CONCLUSIONSA novel mutation in the FV gene was identified in the proband with congenital FV deficiency. The mutation was 675delA in exon 4 resulting in a frameshift and a premature termination codon.

Adolescent ; Blood Coagulation ; Factor V ; analysis ; genetics ; Factor V Deficiency ; blood ; genetics ; Female ; Humans ; Mutation

OBJECTIVETo investigate effect of oxidized low-density lipoprotein (ox-LDL) on memory CD8T cell subpopulation differentiation in mice with autoimmune diabetes.

METHODSCultured splenic CD8T cells from pre-diabetic NOD mice isolated with magnetic beads were treated with 30 µg/mL ox-LDL and 10 U/mL interleukin-2 (IL-2) for 24 h and the control cells were treated with IL-2 only. Flow cytometry was used to determine the percentage of splenic CD8IFN-γT cells, expressions of CD8, CD44 and CD62L on the T cells, and the activation of T cell factor-1 (TCF-1) and STAT-3. The CD127memory T cells were purified and transplanted into the pre-diabetic NOD mice via the tail vein, and the blood glucose was recorded weekly and survival time of the mice was monitored.

RESULTSTreatment with ox-LDL significantly reduced islet β cell-specific cytotoxic CD8T cells as compared with the control group [(0.7∓0.03)% vs (2.7∓0.14)%, P<0.01]. The percentage of effector memory CD8T cells (Tem) in the total memory CD8T cells was reduced [(10.3∓0.71)% vs (30.3∓1.36)%, P<0.01] and that of stem cell-like memory T cells was significantly increased [(72.3∓3.8)% vs (55.1∓2.61)%, P<0.05] following ox-LDL treatment, which also resulted in significantly decreased activation of TCF-1 [(14.5∓0.82)% vs (34.2∓1.23)%, P<0.01] and pSTAT-3 [(3.3∓0.12)% vs (22.1∓1.1)%, P<0.01]. Transplantation of ox-LDL-treated memory T cells in pre-diabetic NOD mice obviously inhibited the increase of blood glucose and prolonged the survival time of the mice (P<0.05).

CONCLUSIONOx-LDL decreases the activation of transcriptional factors TCF-1 and phosphorylation of STAT-3, inhibits the formation of effector memory CD8T cells with long-term cytotoxicity, but promote the generation of stem cell-like memory CD8T cells, which result in suppression of islet β cell-specific effector cytotoxic CD8T cell differentiation to lessen autoimmune injury to the islet β cells.

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

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