Child, Preschool ; Humans ; Hypercalcemia ; etiology ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

Objective To learn about electrolytes imbalance and water intoxication in children treated with high-dose cyclophosphamide(HD-CTX)as well as the renal function and the relative clinical symptoms,and study the mechanisms of hyponatremia.Methods Patients' clinical manifestations during and after HD-CTX therapy were summarized.Serum electrolytes and creatinine(Cr)were detected before and 6 or 8 hours after therapy with HD-CTX,antidiuretic hormone(ADH) in some patients were measured.Results Of 108 therapeutic cases 24 accompanied with vomits and 22 with a decreased urine output,in which 4 developed eyelid or ankle edema.Seven cases had neural-sarcous symptoms and 5 cases had abdominal pain or diarrhea.Serum sodium decreased significantly after HD-CTX[(139.12?3.30) mmol/L vs(134.06?8.23) mmol/L] in whom rechecked after 6 h,(141.77?3.59) mmol/L vs(133.26?6.41) mmol/L in those rechecked after 8 h(Pa0.05].Serum Cr increased 8 h after therapy[(29.95?13.61) ?mol/L vs(43.33 ? 17.25) ?mol/L P

0.05)],which less than in control group [(4.614?1.683) IU/cm,(0.119?0.068) IU/cm,(564.2?53.8) ?m Pa

MicroRNA (miRNA) is an endogenous, non-coding single-stranded RNA that regulates a variety of signal pathways or cytokines.Recent studies have confirmed that miRNA can affect alkaline phosphatase activity and matrix mineralization in bone formation, and plays an important role in osteogenic differentiation and cartilage differentiation.Abnormalities in the osteogenesis process can lead to osteogenesis imperfecta, Feingold syndrome, and femoral head necrosis.This review summarizes the specific mechanism of osteogenic differentiation and cartilage differentiation regulated by miRNA, suggesting the new clue for the future research about the underlying mechanism of bone development and clinical treatment of bone dysplasia from epigenetics.

A pairs of primers were designed according to the fusion protein(F)gene sequences of canine distemper virus(CDV)in GenBank.A 369 bp fragment aimed signal peptide fragment of F gene was amplified.The PCR products from viscera samples,blood,urine of fur animals including foxes,minks and raccoon dogs,which collected in the years 2005-2007,were cloned to pMD18-T Vector and sequenced.We obtained 13 positive signal peptide fragments from wild-type strains.The results indicated there was obviously genetic diversity between the wild-type strains and CDV3 and other vaccine strains.The homology with CDV3 is 80.7%-83.2%in nucleotide,and 64.8%-71.3%in amino acid.The analysis for the hydrophobic regions indicated the function of signal peptide fragment may be changed.This study can offer aca- demic data to research of CDV genetic variation and epidemiology.

Objective To explore the siRNA as a new antiviral therapy,evaluate the inhibition effect of siRNA based on vector on the HBV of HepG2.2.15 cell,and observe the side effect and toxicity of siRNA vector on cells and the off-target effect of siRNA.Methods Three pairs of siRNA duplexes targeting HBV C gene were designed as double strands,and the duplex were annealed and ligated into the p-Silencer-Cmv 4.1-hygro vector.The ligation products were used to transform JM109 cells.The clones with shRNA were obtained,and the vectors were purified.After the initial identification of the vector with agarose gel and the size of the inserted sequence got examined by native polyacrylamide gel electrophoresis,furthermore the sequencing was further carried out.The recombinant plasmids were purified with ultrapure Midipreps DNA Purification System.Then HepG2.2.15 cells were transfected with the plasmid mixed with siPort XP-1.The expression of HBsAg and HBeAg were detected by immunofluorescence and Western blot,and the HBV RNA was investigated by RT-PCR.Furthermore the real-time quantitive PCR was carried out to detect the changes of HBV DNA.In order to evaluate the toxicity of the shRNA,MTT was used to examine the growth rate and curve of cells.The ELISA was performed to detect the changes of interferon-? (IFN-?).Results The Western blot showed that the HBsAg and HBeAg protein were suppressed with (81.15?0.69)%,(88.12?0.92)% respectively by vector p-C2 on the third day of post-transfection.It had the similar result indicated by immunofluorescence.And the RT-PCR showed that the specific siRNA targeting HBV C gene could markedly suppress the expression of HBV mRNA and the HBV C gene mRNA was inhibited with 96.9%.The real-time quantitive PCR showed that the specific functional siRNA could markedly suppress HBV DNA copy with two orders of magnitude,while the siRNA vector had no effect on the growth of cell showed by MTT detection.Compared with the non-transfected group and p-NC group,the IFN-? level was almost the same with siRNA p-C1,p-C2,p-C3 groups.Conclusions The siRNA based on the expression vector can suppress the expression and replication of HBV in HepG2.2.15 cell.The inhibition effect was specific and had a certain dependency on siRNA concentration.No toxicity effect was found in the study.And the drug resistance wouldn′t happen because the silence was based on the split of gene.

OBJECTIVEWe identify ionizing radiation-induced mitochondrial DNA (mtDNA) deletions in human lymphocytes and their distribution in normal populations.

METHODSLong-range polymerase chain reactions (PCR) using two pairs of primers specific for the human mitochondrial genome were used to analyze the lymphoblastoid cell line following exposure to 10 Gy (60)Co γ-rays. Limited-condition PCR, cloning and sequencing techniques were applied to verify the mtDNA deletions detected with long-range PCR. Human peripheral blood samples were irradiated with 0, 2 and 6 Gy (60)Co γ-rays, and real-time PCR analysis was performed to validate the mtDNA deletions. In order to know the distribution of mtDNA deletions in normal population, 222 healthy Chinese adults were also investigated.

RESULTSTwo mtDNA deletions, a 7455-bp deletion (nt475-nt7929 in heavy strand) and a 9225-bp deletion (nt7714 -nt369 in heavy strand), occurring between two 8-bp direct repeats, were identified in lymphoblastoid cells using long-range PCR, limited-condition PCR and sequencing. These results were also observed for (60)Co γ-rays irradiated human peripheral blood cells.

CONCLUSIONTwo novel mtDNA deletions, a 7455-bp deletion and a 9225-bp deletion, were induced by ionizing radiation. The rate of the mtDNA deletions within a normal population was related to the donors' age, but was independent of gender.

Cell Line ; Cloning, Molecular ; Cobalt Radioisotopes ; DNA Damage ; genetics ; radiation effects ; DNA, Mitochondrial ; genetics ; radiation effects ; Gene Deletion ; Genetic Markers ; Humans ; Lymphocytes ; radiation effects ; Radiation, Ionizing ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the influence of the polluted SY River on children's growth and sex hormones, and provide scientific data for assessment of the polluted status of the SY River.

METHODSThe study areas were selected randomly from the SY River Basin. Lead (Pb), mercury (Hg), arsenic (As), phthalates (DEP, DBP, DMP, DEHP), and bisphenol A (BPA) were measured both in the river water and in the drinking water. School children were selected by cluster sampling (n=154). Physical development indexes (height, weight, bust-circumference, and skinfold thickness) and sex hormones [testosterone (T) and estradiol (E2)] were measured for all the children.

RESULTSThe contents of Pb and Hg exceeded Class V standards of surface water quality in each section of the river and other indicators exceeded Class III. Compared to the control area, the concentrations of Pb, Hg, As, BPA, DEP, and DBP in the drinking water were significantly higher than in the polluted area (P<0.05). Children from the control area had significantly lower E2 and T than children from the polluted area (P<0.05). Among anthropometric results, only skinfold thickness had statistically significant difference between the two groups (P<0.05), while the other indexes showed no significant differences between the two groups (P>0.05).

CONCLUSIONThe drinking water has been polluted by the SY River and affected serum sex hormone levels of children living in the polluted area.

Adolescent ; Adolescent Development ; drug effects ; Child ; Child Development ; drug effects ; China ; Female ; Gonadal Steroid Hormones ; metabolism ; Humans ; Male ; Rivers ; chemistry ; Water ; chemistry ; Water Pollutants, Chemical ; toxicity ; Water Pollution, Chemical ; adverse effects ; Water Supply ; analysis

OBJECTIVETo screen childhood ALL related microRNAs (miRNAs), analyze association of miRNAs expression profiles with prognosis and relapse in childhood acute lymphoblastic leukemia (ALL) and explore new indicator for predicting relapse and prognosis.

METHODSmiRNAs expression profile was analyzed by gene chip in 49 newly diagnosed childhood ALL and 12 primary immune thrombocytopenia (ITP) cases (as control group). Abnormal expression of miRNAs was verified by qRT-PCR. The correlation of miRNAs expression pattern with indicators predicting early prednisone response and relapse within a year was analyzed.

RESULTSSpecific expression of miRNAs profiles associated with prednisone response and early relapse in childhood ALL was identified. Eight miRNAs (miR-18a, miR-532, miR-218, miR-625, miR-193a, miR-638, miR-550 and miR-633) could distinguish prednisone sensitive from insensitive. The early relapse of newly diagnosed patients with either high-risk or non-high-risk clinical types had some characteristics of abnormal expression of miRNAs, including miR-7, miR-216 and let-7i upregulated, while miR-486, miR-191, miR-150, miR-487 and miR-342 downregulated.

CONCLUSIONSThe initial screening reveals miRNAs differentially expressed from normal in ALL suggesting the potential roles of them in leukemogenesis. MiRNAs expression signatures may be useful for predicting prognosis and relapse in childhood ALL and directing personalized treatment.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; MicroRNAs ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; pathology ; Prognosis ; Recurrence ; Transcriptome

Ovine pulmonary adenomatosis (OPA) is a naturally occurring contagious lung tumor of sheep which was caused by an exogenous retrovirus of sheep, jaagsiekte retrovirus (JSRV). Although no specific circulating antibodies against the virus coud be detected in infected sheep, exogenous JSRV proviral DNA sequences (exJSRV) and JSRV RNA transcripts could be detected in lung tumors, lymphoreticular system and peripheral blood mononuclear cells (PBMC) from sheep affected by OPA. The sheep genome carried 15 to 20 copies of endogenous retrovirus loci (enJSRV) that were similar to JSRV in structural genes but the divergene in U3. Therefore, primers specific for the U3 sequences of exJSRV were designed for the specific PCR and nested PCR (n-PCR). Sensitivity between specific PCR assay and n-PCR assay was compared by using serial dilutions of positive plasmid pJSRV-LTR in a background of 700ng sheep genome DNA. Sensitivity of n-PCR was ten-fold higher than specific PCR. The n-PCR was only available in blood test for detection of JSRV infected sheep and might be useful in epidemiological studies and disease control of OPA.
Animals ; Base Sequence ; Clinical Laboratory Techniques ; DNA, Viral ; analysis ; Endogenous Retroviruses ; genetics ; Jaagsiekte sheep retrovirus ; genetics ; Lung Neoplasms ; virology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Pulmonary Adenomatosis, Ovine ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Sequence Analysis, DNA ; Sheep ; Sheep Diseases ; virology ; Virus Cultivation