Objective To investigate clinical and experimental features of APL with tetraploid clone characterized by double t (15 ; 17). MethodsFive cases of APL with tetraploid clone characterized by double t(15;17) were chosen. Cytogenetic examination of bone marrow was performed with bone marrow or short-period culture. R banding technique was used for karyotype analysis. DNA content in one case was determined by flow cytometry. Immunophenotyping was performed by using a panel of monoclonal antibodies :CD2, CD13, CD15, CD33 and CD34. PML/RARα fusion gene was detected by interphase FISH using dualcolor PML/RARα probe in one case and by RT-PCR in two cases. ResultsAll cases were male with a median age of 38. Their marrow cell morphology examination showed marked hyperplasia with large leukemic cells that had bizarre nuclear configuration. Chromosome analysis revealed that a leukemia clone with tetraploid or near-tetraploid karyotype characterized by double t(15;17) (q22 ;q12) in five cases, of which, one also had a diploid clone with t(15;17) and a normal cell;two had some cells with normal karyotypes.PML/RARα fusion gene was detected by FISH in one of 5 cases and by RT-PCR in 2 of 5 cases. CD33 expression was found in one case. CD13 and CD33 expressions were seen in the other four cases, of which,CD34 or CD2 co-expression was found in one case and in two cases respectively. The result of DNA content showed a single peak which indicated only tetraploid clone whose DNA index was 1. 998 with CV of 8. 2%.All patients obtained complete remission after the treatment with ATRA and/or arsenic trioxide. Conclusions These results indicate that API, patients with tetraploidy and near-tetraploidy have giant and bizarre blasts.Most patients have short-type PML/RARα transcripts. The tetraploidy in APL does not appear to affect the response to treatment of ATRA.

Objective To evaluate the occurrence and management of complications following ultrasound-guided minimally invasive percutaneous nephrolithotomy (MPCNL). MethodsFrom November 2003 to January 2011,2300 cases of ultrasound - guided MPCNL were performed for upper urinary tract stones in our department.Of these cases,renal calculi were found in 1305 cases,upper ureteral calculi in 322,renal and coexisting ureteral stones in 673. Results Among the 2300 cases of MPCNL,a total of 756 (32.9％) patients encountered complications.Of these cases,peel-away sheath placement failure occurred in 184 cases( 8.0％ ),in which six cases needed secondary surgery.Collecting system perforation occurred in 308 cases ( 13.4％ ),fever in 303 cases ( 13.2％ ),including septicemia in 20 cases (0.87％).The 20 septicemia patients received intensive antibiotic treatment and were successfully cured.Thrity-six patients required transfussions due to severe hemorrhaging ( 1.57％ ).Renal vein injury occurred in three cases (0.13％),for which these patients received intensive care therapy to provide haemostasis with a second procedure months later.There was extensive hemorrhage in 16 cases (0.70％) post-MPCNL,super-selective renal artery embolisation was performed in 12 cases and nephrectomy in 1 case.Pleural injury occurred in one case (0.04％) and pleural effusion in two cases (0.09％),all of which were cured with conservative therapy.There were no cases of abdominal organ injury.ConclusionsThe rate of ultrasound guided complications in MPCNL was lower than that of X-ray guided MPCNL in adjacent organ injury,but higher in complications related to the access ( such as:peel-away sheath placement failure,collecting system perforation),parenchymal bleeding and fever.Most complications (i.e.,bleeding,fever) could be managed conservatively or with minimally invasive procedures ( i.e.superselective renal embolisation,antibiotics treatment) when the complications were recognized early.Renal severe hemorrhage in operation,delayed hemorrhage and infection after MPCNL were several of the severe complications that required active prevention and cure measurement.

Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.

Objective To establish a pancreatic ?-cell developmet fish model with specific spatial expression patterns.Methods Molecular cloning,microinjection,whole embryo in-situ hybridization(WISH)and fluorescence microscopy in living were used to analyze of ?-cell development through generation of transgenic zebrafish.ResultsScreened and established pancreatic ? cells of transgenic zebrafish,and confirmed the fluorescence protein expression in the same spatiotemporal pattern with endogenous insulin gene to achieve dynamic monitoring islet ?-cell development situation in vivo.Conclusion The pancreatic ? cells of transgenic zebrafish animal model can successfully trace pancreatic ? cell development.

0.05).Coincidence both of them in subtypes of ADHD diagnosed by 2 different ways were lower than 50% in the 6.0-6.9 and over 10.0 years old groups,but coincidence both of them were higher than 60% in 7.0-7.9,8.0-8.9,9.0-9.9 years old groups.What's more,there were significant differences though ?2 variance analysis in subtypes of ADHD by 2 different ways(Pa

Objective To construct the recombinant yeast expressing PreS2 120-146-hepatitis B surface antigen (HBsAg),and to evaluate the immune effects of whole yeast cells.Methods PreS2 120-146 and HBsAg gene sequence were optimized according to the yeast cell codon preference,and were recombined and cloned into pPIC3.5K yeast expression vector to construct pPIC3.5K/PreS2 120-146 plasmid.After digested and linearized by Bgk Ⅱ restriction enzyme,pPIC3.5K/PreS2 120-146-HBsAg recombinant plasmid was electrotransformed into GS115 strain to screen PreS2 120-146-HBsAg-recombinant Pichiapastoris .The expression of PreS2 120-146-HBsAg was identified by sodium doclecyl sulfate polyacrylamide gel electrophogesis (SDS-PAGE),Western blot and enzyme linked immunosorbent assay (ELISA)analysis. BALB/c mice were vaccinated by inactivated whole recombinant yeast cells expressing target protein. Specific antibodies to HBsAg were detected by ELISA.Cytotoxic T lymphocyte (CTL)response induced by interferon (IFN)-γ was detected by reverse transcription-polymerase chain reaction (RT-PCR)when immune spleen cells of mice were stimulated by CTL epitope on HBsAg.Independent sample t test was used. Results Data of PCR detection,restriction enzyme digestion and sequencing analysis showed that recombinant pPIC3.5K/PreS2 120-146-HBsAg plasmid was successfully constructed.SDS-PAGE,Western blot and ELISA verified the expression of PreS2 120-146-HBsAg in the lysate of the recombinant Pichiapastoris induced by methanol.Levels of specific anti-HBsAg IgG antibodies produced by inactivated yeast cells vaccinated mice were comparable to purified HBsAg immunization (t =0.946,P =0.381 ). Analysis of HBsAg-specific CTL responses revealed that the level of IFN-γwas significantly higher when the immune spleen cells of mice were stimulated by CTL epitope peptides on HBsAg (t =2.305 ,P =0.044).Conclusions PreS2 120-146-HBsAg target protein is successfully expressed by construction of recombinant Pichiapastoris . The specific humoral and cellular immune responses are induced by recombinant whole yeast cells vaccinated mice.

Human ribosomal DNA (hrDNA) targeting vector pHr is a homologous recombinant plasmid for human genome which developed in the State Key Laboratory of Medical Genetics. pHr was used to construct a recombinant plasmid pHr-CMG expressing mda-7/GFP fusion gene and its efficacy in the hepatocellular carcinoma cell line Bel-7402 was investigated. The expression of mda-7/GFP fusion gene was detected by fluorescent microscope, RT-PCR and Western blotting, and its function was detected by cell-cycle analyses, MTT assay and Hoechst33258 staining. The results demonstrated that pHr-CMG vector could express MDA-7/GFP fusion protein effectually and the mda-7 gene could induce cell apoptosis and proliferation suppression in Bel-7402 cell line, which might be caused by the G2/M cell cycle arrest. These results also suggested that human ribosomal DNA targeting vector system and the pHr-CMG vector may be applied in further gene therapy researches for hepatocellular carcinoma.

OBJECTIVESome research has shown that there may be memory/caution (M/C) defects in children with primary nocturnal enuresis (PNE). This study aimed to investigate whether the defects affect the intelligence level and the intelligence structure in PNE children.

METHODSIntelligence tests were performed by means of Wechsler Young Children Scales of Intelligence (C-WISC) in 40 children with PNE and 40 age-matched normal children.

RESULTSThe full intelligence quotient (FIQ), verbal IQ (VIQ) and performances IQ (PIQ) in the PNE group were in a normal range and did not different from the control group. There were significant differences in the scores for digit extent, decipher, knowledge and arithmetics between the PNE and the control groups (P < 0.05). M/C factor in the PNE group was statistically lower than in the control group (93.44 +/-11.27 vs 100.03 +/-11.79; P < 0.05).

CONCLUSIONSThe total intelligence level of children with PNE was normal, but the M/C factor in the intelligence structure had some defects, suggesting that PNE may be related to the abnormity of executive function in the frontal lobe.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; psychology ; Child ; Female ; Humans ; Intelligence ; Male ; Nocturnal Enuresis ; psychology

Objective To explore the effects and safety of hemoperfusion(HP) therapy on tetramine poisoning in infants.Methods Thirty-five infants with tetramine poisoning were divided into two groups: HP group(n=18) and non HP group(n=17).The changes of blood tetramine concentration and clinical symptom improving of both groups after the treatment were observed together with the adverse effects of HP group.Results The average blood tetramine concentration of HP group was higher than that of non HP group(342.2?333.4 vs 117.9?50.8 ?g/L,P

In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.