[Objective] To set up an optimized protocol for aneuploidy detection from single cells through Array- Comparative Genetic Hybridization (CGH).[Method] Two cell lines,trisomy 18 (Tri-18;GM02732,47,XY,+18) and chromosome 4 segment deletion [sDel-4;GM00343,46,XY,4(del) (qter ＞ p14)],were used in the study.In combination of 10 k 2.0 SNP mapping array platform and multiple displacement amplification (MDA),the diagnostic accurate rates of MDA product from single cells of the two cell lines using gDNA and single-cell MDA product as reference were compared.[Result] An extremely lower call rate (3.2 ± 1.2)% in the negative control group was observed compared to the experiment groups.When the single-cell MDA product was used as reference,the standard deviations of Log2 (signal intensity ratio) were significantly decreased in both groups,compared with when the gDNA as reference (P ＝ 0.004).Through CNAT analytic software,some specific chromosomes (16,17,19,and 22) presented obvious preferential amplification (PA) when the gDNA was used as reference,but this PA could be eliminated when single-cell MDA product was used as reference.[Conclusion] 10 k 2.0 SNP mapping array in combination with MDA could be a quick,highly efficient and accurate method to detect aneuploidy in single cells.

Objective To evaluate the success rate, safety and efficacy of endoscopic retrograde cholangiopancreatography (ERCP) after Billroth Ⅱ gastrectomy. Methods Data of 75 patients with biliary disease after Billroth Ⅱ gastrectomy, who underwent ERCP from January 2007 to November 2009, were retrospectively analyzed. Results In 75 patients, afferent loop intubation was achieved in 69 (92%) and selective cannulation of bile duct were successful in 68 (68/69, 98. 5%). Diagnostic procedures were carried out in 3 patients, and therapeutic in 65 others, which included EST plus stone removal and ENBD in 16, ERBD in 19, EMBE in 18 and EBD plus stone removal and ENBD in 12. Afferent loop perforation occurred in 1 patient (1.3%) and was treated surgically, and 2 acute pancreatitis (2. 6%) were treated conservatively.There was no complication of bleeding. Conclusion ERCP after Billroth Ⅱ gastrectomy is safe and efficiency for biliary disease.

Objective To evaluate the effect of anti-RANTES monoclonal antibody in combination with ciclosporin (CsA)upon inhibiting the rejection response during secondary heart transplantation in mouse models. Methods BALB/c mouse models were used as the donors and C57BL/6 mice were utilized to establish secondary heart transplantation recipient models. The animals were randomly divided into the control (physiological saline,n =6 ),A (anti-RANTES monoclonal antibody treatment,n =6 ),B (CsA treatment,n =6 ) and C groups (anti-RANTES monoclonal antibody combined with CsA treatment,n=6). The survival time of heart after secondary transplantation was observed. The degree of acute heart rejection was assessed by histopathological analysis. The relative expression levels of RANTES,interleukin(IL)-2,IL-1 0,interferon(IFN)-γand transcription growth factor(TGF)-βmessenger ribonucleic acid (mRNA)in the heart grafts were quantitatively measured by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The serum levels of RANTES,IFN-γ,IL-2,IL-1 0 and TGF-βwere detected by enzyme-linked immune absorbent assay (ELISA). Results The heart grafts of all mice survived after secondary cardiac transplantation. Compared with the control group,the survival time of hearts in group A,B and C was significantly prolonged (all P<0. 01 ). Pathological staining revealed that the quantity of infiltrated inflammatory cells in group C was significantly decreased than those in the other groups. The expression levels of heart RANTES,IFN-γand IL-2 mRNA in group C were significantly down-regulated, whereas the expression levels of IL-1 0 and TGF-βmRNA were considerably up-regulated compared with those in the other three groups (all P<0. 05). The serum levels of RANTES,IL-2 and IFN-γin group C were significantly down-regulated, whereas the serum contents of IL-1 0 and TGF-βwere considerably up-regulated compared with those in the other three groups (all P<0. 05 ). Conclusions Combined application of anti-RANTES monoclonal antibody and CsA can effectively induce the immune tolerance to secondary cardiac transplantation and prolong the survival time of the cardiac grafts in mouse models.

OBJECTIVETo evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.

METHODSA fibroblast cell line (Tri-18; GM02732, 47, XY, +18) was used as the template, and 6 groups were set up in the study. Groups A and B were positive and negative control, respectively; groups C-F were experimental groups involving the MDA products from 1, 2, 5 and 10 cells respectively. In combination of single nucleotide polymorphism (SNP) array, the product of each group was assessed based on the genome coverage, loss of heterozygosity (LOH) rate and allele dropout (ADO) rate.

RESULTSThe nonspecific product of negative control presented an average call rate of 3.2%. The genome coverage of the MDA product increased from 86.4% to 96.4% with the increasing number of template from 1 to 10 cells, while the LOH rate and ADO rate decreased significantly (P<0.05).

CONCLUSIONMDA is a highly efficient and reliable method for whole genome amplification. The fidelity of MDA will be improved significantly with the increasing number of template cells. 10K 2.0 SNP mapping array is a quick, accurate and comprehensive method to evaluate the fidelity of amplified DNA products, but the ADO SNPs should be distinguished from those of preferential amplification from the LOH loci to avoid errors.

Cell Line ; Cells ; cytology ; DNA ; genetics ; Humans ; Loss of Heterozygosity ; Nucleic Acid Amplification Techniques ; methods ; Polymorphism, Single Nucleotide ; Templates, Genetic

Objective To synthesize and investigate cytotoxicity of an indole-chalcone derivative FC58. Methods The target compound was synthesized through the Aldol condensation with 1-(3,4,5-trimethoxyphenyl)ethan-1-one and 1H-indole-3-carbaldehyde. The Cell Titer-Blue method was used to determine in vitro cytotoxicity. The cell cycle experiment was performed to analyze the action characteristics of FC58. Results FC58 exhibited high cytotoxicity against various leukemia cells and resulted in G2/M phase arrest. It showed stronger drug resistant index than traditional tubulin inhibitors such as paclitaxel, vinblastine and doxorubicin. Conclusion FC58 represents a promising lead compound for multi-drug resistant leukemia.

Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia(ALSP)is a clinically rare autosomal dominant genetic disease,and its specific pathogenesis is not yet clear.The colony-stimulating factor 1 receptor(CSF1R)is a transmembrane tyrosine kinase receptor on the cell surface and mutations in the gene encoding it have been identified as potential pathogenic factors for ALSP.However,the specific mechanisms by which CSF1R gene mutations lead to the onset of ALSP are still unclear.After reviewing the mutation sites and pathogenic mechanisms of CSF1R in the pathogenesis of ALSP,CSF1R mutations have been shown to cause microglial dysfunction through mechanisms such as dominant-negative effects,loss of function,haploinsufficiency,and gain of function,thereby leading to the onset of ALSP.A deeper understanding of the causes of ALSP will help in exploring potential treatment methods.