OBJECTIVETo investigate the relationship between cytochrome P4501A1 (CYP1A1) Msp I gene polymorphism and childhood acute leukemia (AL).

METHODSRelevant literature was extensively searched and screened by Pubmed and Wanfang Database, Chinese Science Journal Database and Chinese Journal Net. Various data consolidation, combined OR values and their 95% CI were tested by RevMan 4.2; Funnel plots were used for the bias analysis.

RESULTSSix related literatures were found to meet the requirements. According to heterogeneity results, there was no significant difference in homozygous types(P>0.05), while there was significant difference in two others types (P all<0.05). For wild CYP1A1MspI homozygous for the reference group, Combined OR of heterozygous mutation, homozygous, heterozygous + homozygous mutation in AL and control groups were 1.18, 0.96, and 1.10 respectively. Subgroup analysis: Z values of CYP1A1MspI homozygous, heterozygous + homozygous in the acute lymphoblastic leukemia (ALL) and the control group were 0.10 and 0.76 respectively, Z values in non-acute lymphoblastic leukemia and control group were 0.74 and 0.75.

CONCLUSIONThere is no correlation between CYP1A1MspI gene polymorphism and the susceptibility of childhood AL.

Acute Disease ; Child ; Cytochrome P-450 CYP1A1 ; genetics ; Genetic Predisposition to Disease ; Heterozygote ; Humans ; Leukemia ; enzymology ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate the toxic and DNA damaging effect of acrylamide (AA) on human keratinocytes and its mechanism.

METHODS(1) After the keratinocyte cell line HaCaT cells were exposed to AA with different concentrations for 44 hours, cell survival rate was detected by MTT method. (2) The effects of DNA damage of exposed cells were detected by comet assay. (3) After treating the cells with 2.00 mmol/L of AA plus 0.50 mmol/L of 1-aminobenzotriazole (1-ABT), an inhibitor of cytochrome P-450 enzymes (CYP-450), for 4 hours, the relationship between DNA damage and CYP-450 was studied.

RESULTS(1) Cytotoxicity measurement of AA showed that cell survival rate decreased significantly after 44-hour treatment. (2) Cytotoxicity was not detected after 4-hour AA treatment, but significant DNA damage was observed in all treatment groups, and the degree of damage increased with the concentration of AA. Moreover, the tail lengths of comet cells were in dose-effect relationship. As for cells treated by 1-ABT with 2 mmol/L AA, comet rate and tail length were 15.4% and (8.2 +/- 2.0) micro m respectively, which were decreased significantly (P < 0.01) when compared with 2 mmol/L AA treatment group [80.6% and (44.3 +/- 4.0) micro m].

CONCLUSIONSAcrylamide has significant cytotoxicity and genotoxicity on HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolite(s) of AA through CYP-450.

Acrylamide ; toxicity ; Cells, Cultured ; Cytochrome P-450 Enzyme Inhibitors ; DNA Damage ; Dose-Response Relationship, Drug ; Humans ; Keratinocytes ; drug effects ; enzymology

OBJECTIVETo develop a quantitative method for methylation analysis of the p16 gene based on mismatch hybridization and chemiluminescence.

METHODSGenomic DNA was modified by sodium bisulfite to convert all unmethylated but not methylated cytosines to uracil, and subsequently a pair of primer having no CpG sites was designed for amplification target DNA containing methylated or unmethylated CpG sites. The PCR product spanning CpG sites were hybridized with two oligonucleotide probes which perfectly matched the methylated and unmethylated CpG sequences respectively, and the hybrids were detected by chemiluminescent method. The percentage of methylated target sequences could be estimated by calculating the ratio of signals obtained with two probes.

RESULTSThe percentage of methylation of artificial mixtures DNA showed a linear relation. There was a negative correlation between the methyaltion index with p16 transcriptional mRNA of p16 gene in tumor cell lines.

CONCLUSIONCompared with existing methods, this assay is nonisotopic, rapid, simple, and can be widely applied to the study of DNA methylation.

Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Genes, p16 ; Humans ; Luminescent Measurements ; Nucleic Acid Hybridization ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfites

OBJECTIVETo study the immunophenotype and its relationship with clinical characteristics in children with acute lymphoblastic leukemia (ALL).

METHODSBone marrow or blood samples (2-3 mL) with heparin anticoagulation from 139 children with ALL were obtained, and immunophenotypes were identified by flow cytometry.

RESULTSIn 139 ALL children, there were 103 cases (74.1%) of B-ALL, 24 cases (17.3%) of T-ALL, 12 cases of T/B biphenotypic (8.6% of T/BALL). In the 103 children with B-ALL, CD19 (90.3%), CD10 (83.5%) and CD20 (27.2%) were expressed as major antigens. In the 24 children with T-ALL, the major antigens were CD3 (79.2%), CD7 (66.7%) and CD5 (33.3%). In the 12 children with B/T-ALL, T-lymphoid antigens included CD7 (50.0%) and CD5 (41.7%), while the B-lymphoid antigens included CD19 (50.0%) and CD10 (33.3%). Of the 139 children with ALL, 32 cases (23.0%) showed myeloid antigen expression (My+ ALL) and the main expression antigens were CD13, CD33, CD14 and MPO. CD34 was expressed in 31 cases. CD34-positive expression (15.6%) in My+ ALL children was significantly lower than in My-ALL children (24.3%). HLA-DR was expressed in 82 of the 139 ALL children. The expression of CD10, CD34 and HLA-DR in the standard-risk, medium risk, high-risk ALL children was significantly different. There were significant differences in gender and incidence of bleeding between the My+ ALL and My-ALL groups (P<0.05).

CONCLUSIONSImmunetyping can differentiate the sources of leukemic cells. The expression of CD10, CD34 and HLA-DR antigen is related to the clinical classification of ALL.

Adolescent ; Child ; Child, Preschool ; Female ; HLA-DR Antigens ; analysis ; Humans ; Immunophenotyping ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology

OBJECTIVETo investigate the biological characteristics of childhood T-lineage acute lymphoblastic leukemia (T-ALL) and their clinical significance.

METHODSImmunophenotyping was performed by three-color flow cytometry analysis using CD45 /SSC gating in 23 children with newly diagnosed T-ALL. Meanwhile cytogenetic analysis was performed.

RESULTSCD3(+) expression of T-lineage antigens was apparently higher than CD7(+) and CD5(+) expression. CD19(+) expression of B-lineage antigens was apparently higher than CD22(+), CD10(+) and CD20(+) expression. Myeloid antigen was expressed in 4 cases (17%). CD34(+) and HLA-DR(+) were observed in 4 cases (17%) and 5 cases (22%), respectively. cCD3(+) and cCD79(+) were expressed in 23 cases (100%) and 22 cases (96%), respectively. The chromosome detection in 8 cases with T-ALL showed hyperdiploid or Ph(+) chromosome (one case each). The fusion gene detection in 5 cases showed MLL rearrangements in two cases and positive SIL/TAL1 fusion gene in one case. CD3 expression was related with the complete remission rate.

CONCLUSIONSImmunophenotyping is an important tool for diagnosis of T-ALL. However, the immunophenotype of T-ALL is heterogeneous. So, immunophenotyping along with cytogenetic and molecular genetic analysis is needed in the treatment and prognosis evaluation of T-ALL.

Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Humans ; Immunophenotyping ; Male ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology

This paper reviewed the worldwide research progresses of the genus Laggera both on phytochemical and pharmacological work in the past few decades. The main secondary metabolites of this genus are proved to be sesquitepenoids, flavonoids and phenolic acids. Phamacological investigations revealed that the certain extracts of some Laggera species possess significant bioactivities on anti-inflammation, anti-tumor and anti-viral infection. This review afforded the comprehensive description of the active components as to provide useful references to elucidate their historical clinical application on upper respiratory infection, influenza, parotitis, and recurrent herpes viral infection.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; therapeutic use ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Antiviral Agents ; pharmacology ; therapeutic use ; Flavonoids ; chemistry ; isolation & purification ; therapeutic use ; Humans ; Influenza, Human ; drug therapy ; Molecular Structure ; Parotitis ; drug therapy ; Phytotherapy ; Plants, Medicinal ; chemistry ; Ranunculaceae ; chemistry ; Respiratory Tract Infections ; drug therapy ; Sesquiterpenes ; chemistry ; isolation & purification ; therapeutic use

OBJECTIVETo investigate the expressions of matrix metalloprotein-2 (MMP-2) and tissue inhibitor of metallopeptidase inhibitor-1 (TIMP-1) in the renal allografts of patients with chronic active antibody-mediated rejection (ABMR), and explore their role in the pathogenesis of ABMR.

METHODSImmunohistochemistry and computer-assisted image analysis were used to detect the expression of MMP-2 and TIMP-1 in the renal allografts of 46 patients with interstitial fibrosis and tubular atrophy (IF/TA), with 15 normal renal tissue specimens as the control. The association of MMP-2 and TIMP-1 with the pathological grade of IF/TA in ABMR was analyzed.

RESULTSThe expressions of MMP-2 and TIMP-1 significantly increased in the renal tissues of the patients as compared with the normal renal tissues (P<0.05). MMP-2 expression tended to decrease, while TIMP-1 and serum creatinine increased with the pathological grades of IF/TA (P<0.05). In IF/TA group, the expression of TIMP-1 was positively correlated to serum creatinine level (r=0.718, P=0.00<0.05).

CONCLUSIONAbnormal expressions of MMP-2 and TIMP-1 can promote the development of renal fibrosis in chronic ABMR.

Adult ; Antibody Formation ; Complement C4b ; metabolism ; Female ; Fibrosis ; etiology ; Graft Rejection ; immunology ; Humans ; Kidney ; metabolism ; Kidney Diseases ; pathology ; Kidney Transplantation ; adverse effects ; immunology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Peptide Fragments ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

OBJECTIVETo study the relationship between human parvovirus B19 infection and childhood idiopathic thrombocytopenic purpura (ITP) by the principle of evidence based medicine.

METHODSPapers related to the relationship between human parvovirus B19 infection and childhood ITP published between 1994 and 2008 were retrieved electronically from the Chinese Journals Full-text Database and the Wanfang Data. These relevant papers on case-control trials were statistically studied by meta analysis.

RESULTSEight papers that met the inclusion criteria were included for this meta analysis. Five hundred and sixteen cases of childhood ITP and 246 healthy controls were enrolled. The meta analysis showed that the incidence of human parvovirus B19 infection in the ITP group was significantly higher than that in the control group (OR=13.71, 95% CI=7.07-26.59, Z=7.75, p<0.01).

CONCLUSIONSHuman parvovirus B19 infection is closely associated with childhood ITP.

Child ; Erythema Infectiosum ; complications ; Female ; Humans ; Male ; Purpura, Thrombocytopenic, Idiopathic ; etiology

BACKGROUND: To mimic physiological and functional requirements of cartilage and subcondral bone, some recent studies have fabricated bilayered scaffolds for osteochondral tissue engineering in cartilage repair. However, in cartilage repair, and little is reported on the use of a single biomaterial instead of composite biomaterials to fabricate bilayered porous scaffolds. OBJECTIVE: To investigate the biocompatibility of an integrated bilayered porous scaffold fabricated with poly(lactic-co-glycolide) (PLGA), and then to discuss its feasibility of constructing tissue-engineered osteochondral graft. METHODS: The integrated bilayered porous scaffolds were fabricated with PLGA by changing the ultrastructure of scaffold.In vitro,bone marrow mesenchymal stem cells(BMSCs)isolated from rabbits were seeded into the bilayered porous PLGA scaffold, co-cultured for 1 week and then observed under scanning electron microscope. After co-culturing 48 hours,the cells were stained by LIVE/DEAD Kit.In vivo,the composite of DiI-labeled BMSCs and the scaffold or the scaffold alone were implanted subcutaneously into the skin of nude mice, and the implants were taken out and stained by hematoxylin-eosin and DAPI staining at 4 and 8 weeks after implantation. RESULTS AND CONCLUSION: (1) We successfully obtained the bilayered porous scaffolds with different pore sizes (the upper layer:100-200 μm and the lower layer:300-450 μm)and 85% porosity.(2)In the in vitro experiment,the LIVE/DEAD staining showed that BMSCs survived well in the scaffold, and the cells adhered well to the wall of pores in all the scaffolds and extracellular matrix deposition was found under the scanning electron microscope.(3)In the in vivo experiment, shown by the hematoxylin-eosin staining, a small amount of chondrocytes formed in the fibrous tissue in the upper layer and a large amount of trabecular bones in the lower layer were found at 8 weeks after implantation in the experimental group. The close integration between the upper layer and the lower layer appeared. BMSCs could survive for 8 weeks in vivo shown by the DAPI staining.However,in the control group,a little fibrous tissue was found without chondrocytes and apparent trabecular bone, and the scaffold was partially degraded. To conclude, the integrated bilayered porous scaffold fabricated with PLGA alone has good biocompatibility, which is feasibly used in osteochondral tissue engineering.

Objective:To investigate the relationship between mitogen-activated protein kinase (MAPK) signaling pathway related signal molecules and the apoptosis of acute promyelocytic leukemia NB4 cells induced by puerariae radix flavones (PRF) and its significance.Methods:The cells were divided into control group [0.025% dimethyl sulfoxide (DMSO) to replace PRF] and 10, 30, 50 μg/ml PRF groups. The proliferation inhibition rate of NB4 cells exposed with PRF for 24, 48 and 72 hours was determined by methyl thiazolyl tetrazolium (MTT) method, and the nuclear morphology was determined by confocal laser scanning microscope after 48 hours. NB4 cells were divided into control group (adding 0.025% DMSO) and 10, 30 and 50 μg/ml PRF with or without 10 μmol/L c-Jun N-terminal kinase (JNK) inhibitor (SP600125) group, and the cells were treated for 48 hours and the changes in the expressions of MAPK pathway related proteins JNK, tumor necrosis factor α (TNF-α), extracellular signal-regulated kinase (ERK) and p38 MAPK were tested by Western blot.Results:10, 30 and 50 μg/ml PRF inhibited the proliferation of NB4 cells in 24, 48 and 72 hours, which was in time- and dose-dependent manners (all P < 0.05). The half-maximal inhibitory concentration (IC 50) at 24, 48 and 72 hours were (40.03±2.23) μg/ml, (22.92±1.72) μg/ml and (17.99±1.48) μg/ml, respectively. The confocal laser scanning microscope showed that NB4 cells displayed distinct apoptotic characteristics after PRF treatment. After co-cultivating NB4 cells with 10 μmol/L SP600125 and different concentrations of PRF for 48 hours, the expression of JNK1 in NB4 cells was suppressed ( P < 0.05), and the expressions of JNK2/3 and p38 MAPK decreased, but the differences were not statistically significant (both P > 0.05). The expressions of ERK1 and ERK2 gradually increased in the single-drug group, while the expression in the combined drug group decreased. The expression of TNF-α in the 50 μg/ml PRF+SP600125 group was down-regulated compared with the 50 μg/ml PRF single-drug group, while the expressions in the 10 and 30 μg/ml PRF+SP600125 groups were up-regulated compared with the 10 and 30 μg/ml PRF single-drug groups. Conclusion:10-50 μg/ml PRF may activate the MAPK signaling pathway through TNF-α. JNK, ERK1/2 and p38 MAPK interact with each other to activate pro-apoptotic related proteins and induce NB4 cells apoptosis.