Acute lymphoblastic leukemia (ALL) is a heterogeneous disease affected by patient' s age,immunologic subtype,genetic and molecular features.In the 54th ASH annual meeting,lots of new discoveries about the biology,prognosis and treatment of ALL were reported.Here highlights of genomic profiling,new targeted therapy and hematopoietic stem cell transplantation for ALL presented in this meeting are introduced.

Aim To illustrate the actions and molecular mechanisms of interventions taken to convert the metabolism pathways of cellular apoptosis caused by hypoxia and hypoxia-reperfusion in hypertrophic cardiomyocytes.Methods Angiotensin Ⅱ(0.1 ?mol?L-1)was applied to induce the hypertrophy of mice cardiomyocytes.The cardiomyocytes received the treatment of hypoxia-reperfusion in a tri-gas incubator to simulate the conditions of hypoxia-reperfusion.Before hypoxia/reperfusion,no drug intervention and pre-treatments of DCA(1 mmol?L-1),TMZ(5 ?mol?L-1),LC(50 ?mol?L-1)and AA(10 ?mol?L-1)were given respectively.The glucose and fatty acid oxidative metabolism rates were measured with radioactive counting methods.RT-PCR and Western blot methods were employed respectively to measure the mRNA and protein expression levels of cytochrome C and apoptosis inducers.The spectrophotometry method was used to measure the activity of Caspase-3 and Hoechst 33258 staining to quantify the percentage of cellular apoptosis.Results At post-hypoxia 12 h and post-reperfusion 4 h,the glucose oxidative metabolism rates in hypertrophic cardiomyocytes all decreased while the fatty acid oxidative metabolism rates increased.DCA,TMZ and LC all could inhibit both the reduction of glucose oxidative metabolism after hypoxia-reperfusion and the elevation of fatty acid oxidative metabolism after hypoxia-reperfusion.AA drove the reduction of glucose oxidative metabolism rate even lower and the fatty acid oxidative metabolism rate even higher in hypertrophic cardiomyocytes after ischemia/reperfusion.At the same time,DCA,TMZ and LC could inhibit the expression levels of mitochondrial cytochrome C and AIF mRNA and proteins,the nuclear translocation of cytochrome c and AIF proteins and the activity of caspase-3.And with the opposing actions to AA,DCA,TMZ and LC could inhibit the apoptotic rate of hypertrophic cardiomyocytes after hypoxia-reperfusion.And AA had the opposite effect.Conclusion Intervening in the metabolism pathway of hypertrophic cardiomyocytes was an effective way to prevent and control their programmed death through inhibiting the expression of mitochondrial apoptotic proteins.

Objective To study the active constituents of Physalis alkekengi var. franchetii MethodsThe compounds were separated by silica gel and Sephadex LH-20 chromatography method, their structures were identified on the spectral analyses and physical data. Results Eleven compounds were isolated and identified as ?-sitosterol (Ⅰ), physalin A (Ⅱ), physalin B (Ⅲ), physalin O (Ⅳ), physalin L (Ⅴ), physalin M (Ⅵ), daucosterol (Ⅶ), ombuine (Ⅷ), 5, 4′, 5′-trihydroxy-7, 3′-dimethoxy-flavonol (Ⅸ), luteolin (Ⅹ), and luteolin-7-O-?-D-glucopyranoside (Ⅺ). Conclusion Compound Ⅸ is a new compound named physaflavonol.

Objective To investigate the sensitivity of BIOMED-2 primer system in adult acute lymphoblastic leukemia (AIJ,) patients Ig gene rearrangement, and to analyze their frequency, corearrangement pattern, utilization of V, D and J genes and composition of junctional regions. Methods Amplification of rearranged IgH (complete and incomplete), IgK, IgK-Kde and IgL was performed in standard PCR in 29 adult ALL patients. Monoclonal PCR products were subjected directly to DNA sequencing. Sequences were identified by comparison with all known human Ig germline sequences to analyze the recombination patterns, somatic mutations and germline gene segments usage. Results IgH, incomplete IgH, IgK, igK-Kde and Igl, rearrangements were found with positive rate of 70.8%, 12.5% , 29.2% , 25.0% and 0 of B-ALL patients, respectively. All B-ALL patients displayed at least one pattern of Ig gene rearrangements. In TALL, one of five patients was found with incomplete IgH rearrangement, two patients were found with IgK rearrangements and two patients were PCR-negative. The sequence analysis showed that the most frequently used V, D, J segments in adult B-ALL patients were from VH3/VH4 families, DH3 family and JH6 family, respectively. Four of five IgK rearrangement used VκI family. 23.5% B-ALL IgH contained scattered replacement mutations with replacement to silent substitution ratio < 1 in complementarity determining regions. Conclusion BIOMED-2 multiplex PCR analysis strategy is a reliable and useful technique in the adult BALL patients.

Objectives To study the FLT3 gene expression and its internal tandem duplication(ITD) mutation in acute nonlymphoblastic leukemia(ANLL) patients.Method Polymerase chain reaction was used to detect the FLT3/ITD mutation in 71 ANLL patients.Results The expression of FLT3 gene were detected in 65/71(91 5%) ANLL cases and ITD mutation was found in 17 cases. 23 9% of ANLL cases were found with FLT3/ITD mutation. The leucocyte count and the percentage of bone marrow blast cells were higher in ANLL patient with FLT3/ITD mutation than that in the ANLL patient without FLT3/ITD mutation (P

ObjectiveTo evaluate the diagnostic value of ultrasound or CT guided percutaneous Trucut needle biopsy on the diagnosis of pancreatic tumors.Methods One hundred and twenty-four patients clinically diagnosed as pancreatic cancer without pathological diagnosis underwent percutaneous pancreatic biopsy by using Trucut needle under ultrasound or CT guidance.ResultsOne hundred and nine procedures of ultrasound-guided biopsy and 15 procedures of CT-guided biopsy were performed,and one patient received 2.3times of punctures.Tissue samples were obtained in all 124 patients,the diagnostic accuracy was 95.2％,among them 115 were adenocarcinoma,5 were cystadenoma,2 were metastasis cancer,1 was cancer of unknown origin and 1 was normal.The sensitivity,specificity,and accuracy were 99.2％ 100％,and 99.2％,respectively.Transient serum amylase increase was observed in 3 patients; 5 patients' abdominal pain aggravated,but all recovered with conservative management.One patient was found to have tumor seeding on the spot of insertion after 34 days.No other major complications occurred.ConclusionsUltrasound or CTguided percutaneous pancreatic 16 ～ 18G Trucut needle biopsy is a safe and simple procedure with excellent diagnostic value for pancreatic cancer.

Objective To set up a multiplex real time quantitative PCR method to detect the expression of WT1 and MDR1 gene simultaneously in acute leukemia patient. Methods Total RNA was extracted from k562 cell line and was reverse transcribed to cDNA by the outer primers of WT1 and MDR1 respectively. The cDNA of WT1 and MDR1 were purified and digested by Bam H Ⅰ and Bgl Ⅱ , and then the two fragments were ligated to form the recombinant fragment WT1 + MDR1. The outer forward primer of WT1 and outer reverse primer of MDR1 were used to amplify the recombinant fragment WT1 + MDR1. The PCR product was purified and cloned into pMD18-T vector, and then transferred into E. coli DH-5α. A new kind of WT1-MDRl-contained standard plasmid was obtained from the positive colony. The recombinant plasmid was verified by digestion with restriction enzyme and PCR amplification. A multiplex real time quantitative PCR method was set up with FAM-labeled MDR1 probe and VIC-labeled WT1 probe in one reaction tube. The WT1 and MDR1 gene expression was detected in forty-seven AL patients and thirty-two controls by this method. Seven patients were followed-up to elucidate the relationship between the gene expression levels and clinical prognosis. Results The recombinant plasmid was confirmed by EcoR1 digestion and PCR amplification. The multiplex real time quantitative PCR technique could reach the sensitivity of WT1 and MDR1 gene up to 102 copy/μl. The standard curve slopes were 0. 999 and 0. 998. The WT1 [ 37 000( 163-6 370 000 )copies/μg RNA ] and MDR1 [ 76 200( 179-18 000 000 )copies/μg RNA ]expression levels of AL patients were significantly higher as compared to the controls [ 258( 0-643 ) copies/μg RNA and 333( 0-779 )copies/μg RNA ]( Z= 6. 755,6. 736, P ＜ 0. 01 ). Following up seven patients with similar regimen of chemotherapy, the WT1 and MDR1 expression correlated to the clinical course. Three AL patients with WT1 and MDR1 expression levels (2 170 and 86 900, 1 130 and 5 860, 1 170 and 586 copies/μg RNA )significantly decreased after chemotherapy and kept in the low range ( 370 and 560,138 and 980, 150 and 690 copies/μg RNA ), and had a favorable outcome. Three AL patients with WT1 and MDR1 expression levels ( 1 600 and 11 800, 24 800 and 968, 48 200 and 1 100 000 copies/μg RNA )decreased after initial chemotherapy, but increased significantly afterwards (20 314 and 25 660,184 364 and 31 530, 15 680 and 878 000 copies/μg RNA ),and suffered clinical relapse. One patient with high WT1 and MDR1 expression levels ( from 81 600 and 1 200 000 copies/μg RNA to 124 100 and 7 632 400 copies/μg RNA )showed the persistence of disease. Conclusions A multiplex real time quantitative PCR method to detect WT1 and MDR1 gene simultaneously is constructed successfully. The expression of WT1 and MDR1 may provide useful information for AL patients prognosis.

Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.

To observe the effects of Yiqi Huayu Decoction, a compound traditional Chinese herbal medicine, on brain-derived neurotrophic factor (BDNF) expression in lumbar nerve root and soleus of rats after lumbar nerve root compression injury.