This paper introduces cytogenetics in myelodysplastic syndrnme(MDS)in three parts:(1)the frequency rate and types of clonal chromosomal abnormalities in MDS;(2)the value of cytogenetics in the diagnosis,classification,prognosis and treatment of MDS;(3)evaluation of the cytogenetics methods in MDS.Every patient suspected of MDS should receive chromosome examination.About 40%～70% of primary MDS patients and 95% of therapy-related MDS(t-MDS) patients have a clonal chromosomal abnormalities.The frequency rate is related to not only the disease's stages,but also the methods of examination.The types of chromosomal aberrations show huge heterogeneity including various numerical and structural abnormalities,the most common of which are numerical abnormalities and deletion.International score system for evaluating prognosis in MDS has been used worldwide.It dividing the prognosis of MDS into three deferent kinds:high risk,low risk and intermediate risk.The conventional karyotypic analysis (CC) is the most useful method.Fluorencence in situ hybridization (FISH) is an important supplement to CC.CC in combination with FISH will be likely to make an accurate chromosomal analysis for most MDS patients.

Objective To evaluate the efficacy of dexmedetomidine mixed with ropivacaine for transversus abdominis plane (TAP) block combined with general anesthesia in the patients undergoing gynecological laparoscopy.Methods Ninety American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients, aged 20-50 yr, with body mass index of 18-24 kg/m2, scheduled for elective gynecological laparoscopic ovarian cyst resection under general anesthesia, were randomly divided into 3 groups (n =30 each) using a random number table: general anesthesia group (group G) , general anesthesia + ropivacaine for TAP block group (group G+R), and general anesthesia + dexanedetomidine mixed with ropivacaine for TAP block group (group G+DR).After the end of anesthesia induction, ultrasound-guided left-sided unilateral TAP block was performed in G+R and G+DR groups.In G+R and G+DR groups, 20 ml of 0.375％ ropivacaine and 20 ml of 0.375％ ropivacaine mixed with 1 μg/kg dexmedetomidine were injected, respectively.After induction of general anesthesia, the laryngeal mask airway (LMA) was inserted, and the patients were mechanically ventilated in the 3 groups.Anesthesia was maintenaned with iv infusion of remifentanil 0.2 μg · kg-1 · min-1 and propofol 4-8 mg · kg-1 · h-1.Narcotrend index was maintained at 37-46.Only group G received postoperative intravenous analgesia.The occurrence of TAP block-related adverse events was recorded.The emergence time, time for recovery of spontaneous breathing, time for removal of LMA, and Steward score at 5 min after removal of LMA were recorded.The occurrence of postoperative nausea and vomiting, and respiratory depression was recorded.Results Compared with G and G+R groups,the emergence time, time for recovery of spontaneous breathing, and time for removal of LMA were significantly shortened (P＜0.05 or 0.01) , and no significant change was found in Steward score in group G+DR (P＞0.05).There was no significant difference in the above parameters between group G and group G+R (P＞0.05).Compared with group G, the incidence of postoperative nausea and vomiting was significantly decreased in G+R and G+DR groups (P＜0.05).VAS score was maintained ≤ 3 after operation in the 3 groups.No TAP block-related adverse events were found in G+R and G+DR groups.Conclusion Dexmedetomidine mixed with ropivacaine provides faster recovery from anesthesia, and avoids postoperative hyperalgesia when used for TAP block combined with general anesthesia in the patients undergoing gynecological laparoscopy.

Objective To report a rare case of M3r subtype of acute promyelocytic leukemia (APL)with 3'-end of RARα (3'RARα) submicroscopic deletion, and the characters of morphologic, cytogenetic,molecular genetic and molecular biology studies. Methods Chromosomes of bone marrow (BM) cells were prepared with direct method and short-term culture method, and R-banding technique was used for karyotypic analysis. Fluorescence in situ hybridization (FISH) assays were performed on fixed BM cells using the following specific DNA probes: CEP X/Y alpha satellite DNA probe, LSI PML-RARα dual-color dual-fusion and LSI RARα dual-color break apart probes. A quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) was performed to detect the PML-RARα transcript. A multiplex nested RT-PCR was also performed, which may simultaneously detect the fusion genes derived from 29 chromosomal aberrations in acute leukemia including PML-RARα, PLZF-RARα and NPM-RARα fusion transcripts. Results R-banding analysis revealed a karyotype of 45,X,-Y[6]/46,XY[8], FISH using CEP X/Y probe further confirmed Y-chromosome loss. FISH analysis with RARα dual-color break apart probe demonstrated a deletion of the entire 3'-end of one allele of RARα gene. Cytogenetic, FISH and RT-PCR analyses showed no PML-RARα,PLZF-RARα, NPM-RARα, NuMA-RARα and STAT5b-RARα rearrangements. Conclusion A new RARαrearrangement involving 3'RARα submicroscopic deletion in APL without X-RARα fusion has been identified.FISH analysis with RARα dual-color break apart probe is a useful method for characterization of this abnormality, but its molecular consequences remain to be elucidated.

Objective To explore the minimal residual disease (MRD) and cellular chimerism in patients with hematopoietic malignancies after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods From May 2001 to June 2005,83 patients received allo-HSCT,including 55 males and 28 females. Of them 49 patients received sibling HLA-matched bone marrow transplantation (BMT),3 HLA-matched peripheral blood stem cell transplantation,8 un-related BMT,9 nonmyeloablative stem cell transplantation (NST) and 14 related haploidentical transplantation. Among them,49 patients were diagnosed as having CML,16 having AML,16 having ALL,one having multiple myeloma and one having malignant lymphoma. Chimerism and MRD were monitored by fluorescence in situ hybridization (FISH) using X and Y specific centromeric probes or gene probes for BCR/ABL,MLL and AML1/ETO. 1000 cells were analyzed for each sample.Results Among 19 patients receiving sex-matched transplant,the former chromosome rearrangements were not found in 16 patients after transplantation,MRD was detected in 10 % of cells in one patient and 1 % of cells having MRD in 4th month after the reduction of immunotherapy,and the patients were still in remission one year after transplantation. Two patients were found having the former chromosomal rearrangement 1 and 4 months after transplantation,respectively,who did not achieve remission after chemotherapy. Over 99 % donor chimerisms were found in 50 patients on day 25,donor cells were at a low level ( 96.2 % ~ 98.7 % ) in 7 patients on day 25,and increased over 99 % later. They were in remission without relapse. The donor chimerisms were decreased gradually in other 7 patients,of them 3 patients with the host cells above 10 % showed hematologic relapse. Four patients with the host cells between 2 %~5 % had different outcomes: 2 patients died of severe GVHD after the reduction of cyclosporine A,one patient got a donor chimerism over 99 % after reduction of immunotherapy,and one patient was still in complete remission.Conclusions FISH could play a pivotal role in the detection of MRD and chimerism. It is helpful to the evaluation of graft and relapse and to the guide of intervention of early immunotherapy.

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.

AIM: To investigate the effects of interferon-alpha (IFN-?) on the development of dendritic cells (DCs) derived from bone marrow mononuclear cells in patients with chronic myeloid leukemia (CML). METHODS: Bone marrow mononuclear cells from 12 CML patients were cultured initially using cytokines as follows: recombined human granulocyte-macrophage colony stimulating factor (rhGM-CSF) plus IFN-? (IFN-?-DCs); rhGM-CSF plus recombined human interleukin-4 (rhIL-4) (IL-4-DCs); IFN-? alone; rhGM-CSF alone in 10% FBS RPMI-1640 medium for 7 days and then recombined human tumor necrosis factor-? (rhTNF-?) was added for another 3 days. The morphologic features were observed by Wright's staining under inverted microscope. CD_ 80,CD_ 86,CD_ 83,CD_ 1a and HLA-DR expression were assayed by flow cytometry. Cytogenetic analysis was performed for one CML patient by fluorescence in-situ hybridization (FISH), and the functions of antigen presenting were tested by mixed lymphocyte reaction (MLR). RESULTS: IFN-?-DCs displayed features in morphology that was similar to those of IL-4-DCs with delicate membrane projections. IFN-?-DCs showed an increase in expression of CD80, CD86, CD83, HLA-DR and more intense abilities of allogeneic antigen presentation with and without rhTNF-? stimulation, compared with the control groups of IL-4-DCs. FISH confirmed the DCs of both groups were leukemic origin. CONCLUSIONS: (1) IFN-? promoted the differentiation/activation of bone marrow mononuclear cells from patients with CML into activated dendritic cells. (2) The phenomenon of generation of activated DCs in vitro might contribute to therapeutic effect of IFN-? in CML. (3) IFN-? may be valuable for the generation of active bone marrow mononuclear cells-derived DCs to be as vaccination strategies of CML patients.

Objective To understand healthcare-associated urinary tract infection (HA-UTI)and pathogens causing HA-UTI in intensive care unit (ICU)patients,so as to provide scientific basis for the prevention and control of HA-UTI. Methods Targeted surveillance data about HA-UTI in 32 hospitals in 2013 were analyzed.Results A total of 23 680 ICU patients were monitored,157 cases of HA-UTI occurred,HA-UTI rate was 0.66%;the usage rate of urinary tract cathe-ter was 80.83%,catheter-associated UTI was 1.25‰.A total of 162 pathogenic strains were detected,the percentage of fungi,gram-negative bacteria,and gram-positive bacteria was 40.74% (n=66);31.48 % (n=51),and 27.78% (n=45)respectively.Conclusion The main pathogens causing HAI-URI are fungi,comprehensive intervention measures should be taken to control HA-UTI in ICU patients.

Objective To understand the molecular aberration at whole genomic level,CGH (comparative genomic hybridization) was used to investigate genetic abnormality in lung cancer.Methods Comparative genomic hybridization was performed in 17 cases to detect the global genomic aberration in cancer tissue cells.Results All of 17 cases detected by CGH showed chromosomal aberrations.The average numbers of chromosomal gains and losses in each case were 7.0 and 4.8 in NSCLC and 8.4 and 9.6 in SCLC,respectively.The frequency of gains and losses on chromosome had no significant differences between NSCLC and SCLC.The frequencies of gains on chromosomal arms 3q24 -28 and 11q13(58.3％ and 58.3％ ) in NSCLC were significantly higher than that in SCLC(0％ and 0％ ) ( P ＜0.05 and ＜0.05,respectively).Conclusions The cytogenetic aberration generally existed in lung cancer cells.Several regions ( more than one) of chromosomal aberration were involved in the carcinogenesis of NSCLC and SCLC.The regions and frequencies of chromosomal aberration in NSCLC were somewhat different from that in SCLC,which might result in the different biological behavior of the two types of lung cancer.The chromosomal aberration might be served as a marker to differentiate the two types of lung cancer.

Objective To reveal the chromosome abnormalities and their relationship with the clinical phenotype of neonates with congenital malformation.Methods Karyotype analysis of peripheral blood lymphocytes was performed on 396 newborns with congenital malformation,who were recruited at the Children's Hospital Affiliated to Soochow University from Jan.2006 to May 2012,chromosome karyotypes were prepared with neonatal peripheral lymphocytes by conventional G-banding technique.Results 1.Of 396 newborns,159 (40.2％) cases were detected to have chromosomal abnormalities,including karyotype first reported domestically and internationally in 3 cases.2.Trisomy-21 (Down's syndrome),which was the most common abnormal karyotype,was seen in 130 cases,accounting for 81.8％,of whom 119 cases show the standard type,10 cases accompanied by the Robertsonian translocation involving group D or group G,and 1 case accompanied by sexual chromosomal abnormality:inv(Y) (p1 1 q 1 1),+ 21.3.Other common karyotype abnormalities were as follows:del (5) (p 1 2-14) (cats cry syndrome) in 4 cases,trisomy-18 (Edwards syndrome)in 4 cases,45,XO (Turner' s syndrome) in 4 cases,inv (9) (p1 1 q1 2-21) in 4 cases,trisomy-X (super female syndrome) in 1 case,rob(13;14) in 1 case,trisomy-8 in 1 case and del(18) (q22) in 1 case.4.Special faces were seen in 147 cases (92.5 ％),congenital heart disease in 97 cases (61.0％),low birth weight in 72 cases (45.3 ％),congenital anal atresia in 13 cases(8.1％),multiple malformations in 11 cases (6.8％),intestinal malformations in 10 cases (6.2％),extrinsic genital abnormalities in 9 cases(5.7％),meow-like cry in 4 cases(2.5％),limb edema in 4 cases (2.5％),fingers and toe abnormalities in 6 cases(3.6％),congenital brain dysplasia in 6 cases (3.6％),webbed neck in 5 cases(3.1％) and cleft lip and palate in 3 cases(1.8％).Conclusions Chromosomal abnormality is an important factor leading to neonatal birth defects,of which special face,congenital heart disease,low birth weight,and multiple malformations are the main clinical manifestations of chromosomal diseases.

Objective To report two childhood acute myeloid leukemia (AML) patients with t(8;20)(q22;q13) and t(1;8;21)(q32;q22;q22) respectively,as variant t(8；21).Methods Chromosome preparation of bone marrow cells were made using short-term culture and karyotypic analysis was carried out using R and G-banding techniques.Interphase-fluorescence in situ hybridization (I-FISH) and metaphase-FISH (M-FISH) were performed using dual color,dual fusion AML1-ETO probe to detect the AML1-ETO fusion gene.Multiplex RT-PCR was used to demonstrate the expression of AML1-ETO fusion transcript.Results The karyotype of bone marrow cells for these two childhood AML patients were 45,X,-Y,t(8;20)(q22;q13)[12]/46,XY[3](case 1) and 46,XX,t(1;8;21)(q32；q22;q22)[18]/46,XX[2](case 2),respectively.I-FISH and M-FISH confirmed that they all had the AML1-ETO fusion gene and variant t(8;21).The AML1-ETO fusion transcript in both patients was detected by RT-PCR.Conclusion t (8;20)(q22;q13) and t (1;8;21)(q32;q22;q22) are variant t (8;21) in nature.It is important to combine the conventional karyotypic analysis with D-FISH and multiplex RT-PCR to determine the nature and prognosis of AML patients with variant t(8;21).