[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

Objective: To explore the clinical value of prenatal ultrasound in diagnosis of vasa previa. Methods: The images of 65 230 pregnant women who underwent prenatal ultrasound examination were analyzed retrospectively. The accuracy of prenatal ultrasound in diagnosis of vasa previa was calculated. The delivery modes of all pregnant women and outcomes of all perinatal infants with vasa previa diagnosis were followed up. Results: Fifty-three vasa previa cases were diagnosed during the first examination using prenatal ultrasound and 10 cases were eliminated during reexamination in late pregnancy, so 43 cases were diagnosed. Finally 41 cases (41/65 230, 0.06%) were clinically confirmed. The accuracy of middle pregnancy screening (15-28 weeks) and late pregnancy (28-40 weeks) reexamination was 100%(28/28), and the accuracy of late pregnancy was 86.67%(13/15). Cesarean section surgery was performed in all 41 pregnant women, and all fetuses survived. Conclusion: Prenatal ultrasound has a high value of prenatal ultrasound in diagnosis of vasa previa.

Objective To investigate the clinical value of prenatal ultrasound in fetal meningocele and encephalocele.Methods Thirty nine fetuses with meningocele and encephalocele confirmed by prenatal ultrasound and abortion were acquired and their clinical data and sonographic features were retrospectively analyzed.Results Ultrasound screening in 111 620 cases showed 39 fetuses were with meningocele and encephalocele,accounting for 0.35％.Among them,16 cases were diagnosed with meningocele (including 1 case with two bulging parts) and 23 cases with encephalocele.Prenatal ultrasound could clearly detect the location and size fetal skull defect,and bulging features.According to their sonographic features,meningocele or encephalocele was determined.Conclusion Ultrasound could be a reliable prenatal screening method,provide an important basis for clinical intervention and have a significant clinical value in fetal meningocele and encephalocele.

Objective To investigate the role of NRAGE subcellular localization in the EMT and radioresistance of esophageal cancer cells.Methods EMT model cells were established by the treatment of TE13 cells with TGF-β1.To verify the establishment of EMT model and the phenotype of EMT-like TE13R120 cells,EMT marker mRNA and protein were detected by Real-time PCR and Western blot,respectively.Real-time PCR was also used to detect the expression of NRAGE mRNA in three groups.Total NRAGE protein,cytoplasm protein and nuclear protein were measured by Western blot.Results It was found that TGF-β1 could induce morphological alterations of TE13 cells from epithelial to mesenchymal and change the expressions of EMT maker E-cadherin and vimentin (t =13.56,-232.84,P ＜ 0.05),indicating the successful establishment of EMT model cells.Similar expression trends of EMT makers were observed in TE13R120cells (t=15.84,-54.54,P＜0.05).NRAGE mRNA (t=-8.73,-5.62,P＜ 0.05) and total protein in both EMT model cells and TE13R120 cells were higher than that in TE13 cells,especially for the nuclear proteins.However,no differences in NRAGE cytoplasm protein expression were found among the three groups.In addition,there were also no difference of NRAGE mRNA (t =-0.88,P ＞0.05),cytoplasm and nuclear protein between TE13R120 cells and EMT model cells.Conclusions The radioresistant cell line TE13R120 has the EMT-like phenotype that may cause cell radioresistance by changing the subcelluar localization of NRAGE.

OBJECTIVETo investigate the clinical and genetics characteristics of patients with monosomal karyotype acute myeloid leukemia (MK-AML).

METHODSThe karyotypes of 3743 patients with newly-diagnosed de novo AML were analyzed, which had identified 153 cases with MK-AML, for whom the clinical and genetics characteristics were analyzed.

RESULTSThere were 2056 patients (54.9%) among all patients. A total of 153 patients fulfilling the criteria for MK-AML were identified, which comprised 93 males and 60 females, with a median age of 54. The median white blood cell count on presentation was 4.4×10 (9)/L. One hundred and forty-five cases (94.8%) have fulfilled the criteria for complex karyotype (≥ 3 chromosomal abnormalities). Although the monosomy could be found with all autosomes, chromosome 7 has been most frequently involved (38.56%, 59/153).

CONCLUSIONMK-AML is a distinct cytogenetic subtype of AML. Monosomy 7 is frequently detected among MK-AML patients. The monosomal karyotype is common among elder patients with AML.

Adult ; Aged ; Chromosomes, Human, Pair 7 ; genetics ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Monosomy ; Young Adult

OBJECTIVETo explore the clinical and laboratory features of chronic myeloid leukemia (CML) with atypical e14a3 and e19a2 BCR-ABL fusion gene subtypes.

METHODSWe retrospectively analyzed a cohort of CML patients with Ph chromosome positive confirmed by cytogenetic and FISH but classical e13a3(b2a2), e14a2(b3a2)and e1a2 fusion transcripts negative identified by conventional real-time quantification RT-PCR (RQ-PCR). Further RQ-PCR was done with the forward primer and reverse primer designed to detect rare atypical BCR-ABL fusion genes including e14a3 and e19a2 transcripts. Direct sequencing analysis was performed on the PCR products and mutations in the BCR-ABL kinase domain were detected. The clinical data of patients were retrospectively analyzed.

RESULTSSix CML patients were found to carry t(9;22) abnormality and BCR-ABL rearrangement confirmed by FISH but classical BCR-ABL fusion genes negative detected by RQ-PCR. Further RQ-PCR and sequencing analysis confirmed the fusion of BCR exon 14 and ABL exon 3 in five CML patients (case 1-5) and the fusion of BCR exon 19 and ABL exon 2 in one CML patient (case 6). E255K and I293T IM-resistant mutations were detected in case 1 and 2, respectively. Among five cases with e14a3 transcripts, four were CML-CP, one CML-AP. Four patients were male and one was female. The median age was 48 years. The patient (case 6) with e19a2 transcripts was 40-year-old female with a diagnosis of CML-CP and PLT count was more than 1 000×10⁹/L. Imatinib (IM) therapy was administer in case 1, 2, 3, 4 and hematopoietic stem cell transplantation (HSCT) was undergone in case 5 after hydroxyurea (Hu) or interferon failure. Case 1 who had E255K IM resistant mutation, responded poorly to IM but obtained a complete cytogenetic remission (CCyR) after a substitution of dasatinib for IM. Case 2 and 3 achieved CCyR 6 months later after IM treatment and had been maintained well with IM despite I293T mutation in case 2. Case 4 attained CCyR 3 months later after IM treatment but relapsed and died soon. Case 5 was still in CCyR after HSCT. Case 6 with e19a2 transcripts got complete hematologic response after Hu treatment and CCyR was achieved soon after IM therapy.

CONCLUSIONIncidence of CML with atypical transcripts is extremely low. They could benefit from tyrosine kinase inhibitors or HSCT. Rare and atypical BCR- ABL fusion gene subtypes could be missed by conventional RQ-PCR.

Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; classification ; diagnosis ; genetics ; Male ; Middle Aged ; Retrospective Studies

PurposeTo explore the value of prenatal ultrasound in diagnosis of fetuses with limb body wall complex (LBWC).Material and MethodsThe ultrasound images and follow-up results were studied retrospectively in 20 cases of LBWC, and the ultrasonic features of LBWC were analyzed.ResultsAll 20 cases showed rather severe body wall defect and spinal abnormality, including 16 cases of abdominal wall defect, 4 cases of abdominal thoracoabdominal defect, 8 cases of idiopathic scoliosis, 2 cases of kyphosis, 3 cases of twist into a corner, 1 cases of distortion as S shape, and 6 cases of vertebral body arranged in disorder. Moreover, 16 cases presented abnormal umbilical cord,among which 11 had too short umbilical cord, 1 without umbilical cord, and 7 as single umbilical artery (3 cases with short cord). Five cases showed limb abnormalities, among which 2 cases were left lower limb absence, 1 cases as left upper limb absent, 1 cases as left foot varus and right foot slightly varus, and 1 cases as left foot drop.ConclusionPrenatal ultrasonography can accurately diagnose LBWC in time, so as to provide evidence for early clinical intervention.

Objective To evaluate three dimensional fetal heart structure by sono-automatic volume count (SonoAVC) with spatio-temporal image correlation(STIC). Methods The heart volumes datas were acquired by STIC in 32 fetuses with postnatally confirmed diagnoses(20 cases of normal heart, 12 cases of complex congenital heart disease between 20 - 37 gestional weeks), then the volume datas were analyzed offline. SonoAVC software automaticly searched hypoechoic and anechoic structures, and assigned individual colors,and this can be corrected by manual splitting and/or removing and merging of individual segments.Results Individual segments of fetal heat could be separated,and digital casts were generated. The digital casts were obtained successfully in 4cases of transposition of the great arteries(TGA) ,2 cases of tetralogy of Follot,1 case of Ebstein's anomaly, 1 case of double-outlet right ventricle and 20 cases of normal heart.Conclusions Combination of STIC and SonoAVC can demonstrate the size, shape and connection of fetal cardiac cavities and great arteries in three dimensional spatial context. It has the potential both to help in obtaining strctural diagnostic,and to generate 3D visual displays for consultation and teaching.

Objective To Study the effects of high mechanical index ultrasound radiation microbubble on colon cancer cell skeletal.Methods Lovo cell were cultured in vitro,and this study was divided into control group,simple contrast agent group,microbubble plus ultrasound radiation group and simple ultrasound radiation group.Ultrasound contrast agent SonoVue and ultransound (frequency 1.5 MHz and MI 1.7) were used.Results In the control group,the microtubes were densest in dyeing silk,and extended to the edge of a cell.In the ultrasound and microbubble group,the microtubes were weaker and thin,the.network was mainly in the structure of the long axis.The cells arranged in a slight pigmentation to the density cell,to reach out many fine short hair,there were obviously a sense of silk and directional.In the microbubble and ultrasound group,Lovo cells streamed of the central significantly reduced,fluorescent lighting,with short hair.There was no difference in the ultrasound group and the microbubble group with microfilament and microtubule.Conclusions High mechanical index ultrasound radiation can changed into vacant,a trace of the assembly and distributed to the tumor cells are attacking,the transfer of restraining.

OBJECTIVETo investigate whether CpG-oligodeoxynucleotide (CpG-ODN) can improve the detection rate of the karyotypic abnormalities in chronic lymphocytic leukemia (CLL).

METHODSThe bone marrow (BM) or peripheral blood (PB) cells from 57 cases of CLL were collected and cultured with CpG-ODN DSP30+interleukin-2 (IL-2), phytohemagglutinin (PHA), pokeweed (PWM) or IL-2, respectively. Five days later cells were harvested for chromosome preparation. Karyotypic analysis was done using R banding technique. Panel fluorescence in situ hybridization (FISH) was carried out on 19 cases of CLL with normal karyotypes using the following probes: Cen12, D13S25, Rb1, ATM, p53, MYB and IgH. Genomic DNA from 21 cases of them was extracted from BM or PB leukocytes. The immunoglobulin variable heavy chain (IgVH) was amplified by polymerase chain reaction (PCR) and sequenced. CD38 and ZAP70 expressions in the leukemic cells were determined by flow cytometry (FCM).

RESULTSThe detection rate of karyotypic abnormalities in the CpG-ODN+IL-2 group (43.85%) was obviously higher than that in the PHA (15.09%), PWM (17.31%) and IL-2 (3.13%) groups (P<0.01). Fifty-two types of karyotypic abnormalities were found. Among them, trisomy12 (+12) or +12 with other abnormalities were the most common, while translocations were the most frequent structural abnormalities including 3 unbalanced and 11 balanced translocations, among them 7 had rearrangements involving 14q32. Thirteen cases showed one or more abnormalities on FISH including trisomy 12 and p53 deletion each in one case, IgH rearrangement and partial deletion each in one case, 13q14.3 deletion in 11 cases of which 5 cases also had Rb1 deletion, 1 case had Rb1 partial deletion. No case with ATM or MYB deletions was found. PCR detected IgVH mutations in 10/21 cases. FCM showed 10/45 cases were CD38 positive, but 35 /45 were CD38 negative, 11/27 cases expressed ZAP70, but 16/27 did not. Among the 26 cases examined for CD38 and ZAP70 expressions simultaneously, 5 cases were CD38+ZAP70+, 13 were CD38-ZAP70-, 6 were CD38-ZAP70+, and 2 were CD38+ZAP70-, respectively. Statistic analysis showed a correlation between complex karyotype and IgVH without mutation, but no association between karyotype and CD38 or ZAP70 expression was observed.

CONCLUSIONCpG-ODN immunostimulation can obviously raise the detection rate of abnormal karyotypes, especially translocations in CLL. FISH is an important complement to conventional karyotypic analysis. The combination of both methods can provide more comprehensive genetic information for CLL.

Adjuvants, Immunologic ; genetics ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; cytology ; immunology ; Cells, Cultured ; Chromosome Aberrations ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; In Situ Hybridization, Fluorescence ; Interleukin-2 ; genetics ; Karyotyping ; methods ; Leukemia, Lymphocytic, Chronic, B-Cell ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Oligodeoxyribonucleotides ; genetics ; immunology ; Phytolacca americana ; genetics