Surgical Safety of Elderly Hospitalized Patients Stratified by Age in General Surgery / 中国医学科学院学报

Objective To compare the surgical safety of elderly hospitalized patients in different age groups undergoing general surgery,and provide references for preoperative evaluation and treatment decision-making.Methods The inpatients ≥ 60 years old in the department of general surgery were selected from a national multi-center survey conducted from January to June in 2015 and from January to June in 2016.The patient characteristics and postoperative outcomes were described,and the risk factors for adverse postoperative outcomes of patients in different age groups were explored.Results The elderly patients (≥75 years old) accounted for 17.33%.The non-elderly patient (< 75 years old) group and the elderly patient (≥75 years old) group had significant differences in the proportions of patients with three or more chronical diseases (13.18% vs.5.36%,P<0.001),emergency surgery (16.64% vs.7.62%,P<0.001),American Society of Anesthesiologists score≥3 (48.68% vs.27.28%,P<0.001),and postoperative return to the intensive care unit(33.64% vs.12.00%,P<0.001).The occurrence of postoperative infectious complications showed no significant difference between the two age groups (7.29% vs.6.40%,P=0.410),while severe complications differed between the two groups (6.51% vs.2.60%,P<0.001).Besides,emergency surgery was a common independent risk factor for the two age groups.Conclusions Advanced age is not a contraindication to surgery of elderly patients.With consideration to patient's physical conditions and available surgical resources,elderly patients can still benefit from surgery.

Expression of IGLL1 Gene and Its Clinical Significance in Pediatric T-ALL / 中国实验血液学杂志

OBJECTIVE@#To detect the relative expression of IGLL1 (immunoglobulin lambda-like polypeptide 1) mRNA in bone marrow of children with T-cell acute lymphoblastic leukemia (T-ALL), and analyze its correlation with the clinical characteristics and prognosis of the patients, so as to clarify the clinical significance of IGLL1 in pediatric T-ALL patients.@*METHODS@#A total of 56 pediatric T-ALL patients hospitalized in Children's Hospital of Soochow University from June 2012 to December 2017 and treated with CCLG-ALL 2008 regimen were selected. Transcriptome sequencing technology was used to detect the transcription level of IGLL1 gene in children with T-ALL. According to 25% of the IGLL1 transcription level (cutoff value:448), the enrolled children were divided into IGLL1 low expression group (17 cases) and IGLL1 high expression group (39 cases). Combined with clinical data, the correlation between the expression level of IGLL1 and prognosis of the patients was analyzed.@*RESULTS@#The comparative analysis showed that the transcription level of IGLL1 was not correlated with the clinical characteristics of the patients, such as sex, age, bone marrow blast, white blood cell (WBC) count at initial diagnosis. The 5-year OS rate of patients with high IGLL1 expression was significantly higher than that of patients with low IGLL1 expression (76.9%±6.7% vs 47.1%±12.1%, P =0.018). Further comparison of relapse-free survival (RFS) rate between the two groups showed that the 5-year RFS rate of patients with high IGLL1 expression was higher than that of patients with low IGLL1 expression, but the difference between the two groups was not statistically significant (P =0.095). Multivariate COX analysis was conducted on common clinical prognostic factors (age, sex, WBC count at diagnosis, prednisone response on the 7th day, bone marrow response on the 15th day after treatment) and IGLL1 expression level, and the results showed that IGLL1 expression (P =0.012) and prednisone response (P =0.017) were independent risk factors for overall survival in pediatric T-ALL patients.@*CONCLUSION@#In pediatric T-ALL, the OS rate of children with high expression of IGLL1 gene was significantly higher than that of children with low expression of IGLL1 gene, and the expression level of IGLL1 gene was an independent factor affecting the survival of children with T-ALL, which suggests that IGLL1 is a marker of good clinical prognosis of children with T-ALL.

Hypoglycemic Mechanism of Gegen Qinliantang： An Exploration Based on GPR119/cAMP/GLP-1 Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effects of Gegen Qinliantang（GGQL） on the proliferation and apoptosis of intestinal epithelial cells as well as on the expression of cyclic adenosine monophosphate （cAMP）， G protein-coupled receptor 119 （GPR119）， and glucagon-like peptide-1 （GLP-1）， so as to explore its potential hypoglycemic mechanism. MethodTwenty-five Wistar rats were gavaged with GGQL at the dose of 23 g·kg-1 crude drug， twice a day， which meant that 6 mL was administered into each rat per day for preparing the GGQL-containing serum. After seven consecutive times of administration， the intestinal epithelial L （NCI-H716） cells were cultured with different concentrations （1%， 2.5%， 5%， 7.5%， and 10%） of GGQL. The cell proliferation was evaluated using cell counting kit-8 （CCK-8） and the apoptosis by flow cytometry. The GLP-1 and cAMP contents in cell supernatant were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein GLP-1 and GPR119 levels were assayed by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the control group， GGQL significantly reduced the proliferation of NCI-H716 cells（P<0.05）. As the GGQL concentration increased， its inhibitory effect became more obvious. GGQL at each concentration significantly promoted the apoptosis of NCI-H716 cells （P<0.05）. Compared with the control group， GGQL significantly up-regulated the expression of cAMP， GLP-1， and GPR119 （P<0.05）. The results showed that the effect of GGQL was positively correlated with its concentration， and 10% GGQL exhibited the best effect. ConclusionGGQL effectively inhibits the proliferation of NCI-H716 cells and promotes their apoptosis， and it may promote the secretion of GLP-1 by up-regulating the expression of cAMP and GPR119.

Minimally invasive treatment of hallux valgus with bandage for external fixation: Finite element analysis of stability of the osteotomy end / 中国组织工程研究

BACKGROUND: The clinical effect of minimally invasive treatment of hallux valgus is significant. The osteotomy end is stabilized only by external fixation of the bandage. There is currently no research on the stability of the osteotomy end. OBJECTIVE: To study the effect of minimally invasive treatment of the "8" bandage external fixation on the stress and displacement of the osteotomy end in the balanced standing condition after hallux valgus. METHODS: In the minimally invasive treatment of the "8" bandage external fixation finite element model after the hallux valgus operation, three vertical axes (X-axis, Y-axis, Z-axis) were established with the first tibial osteotomy as the center. The X-axis and Y-axis were parallel to the horizontal plane of the foot, pointing to the medial and anterior sides of the foot respectively. The Y axis was perpendicular to the horizontal plane of the foot, pointing upwards. The four nodes defining the distal osteotomy surface were A1 on the upper side, B1 on the outer side, and C1 on the outer side, and D1 on the inner lower side. The proximal end osteotomy surface corresponded to four nodes as A2, B2, C2 and D2. The displacement was positive when it coincided with the direction of the coordinate axis, and negative when it was opposite. Through the finite element analysis, the direction and magnitude of the stress and displacement of the distal and proximal nodes of the osteotomy surface in the balanced standing condition were obtained. RESULTS AND CONCLUSION: (1) The finite element model of the "8" bandage after minimally invasive treatment of hallux valgus was used in a balanced standing condition. The maximum stress at the osteotomy end was at the dorsal side of the osteotomy surface (B2), which was 0.632 MPa. (2) The first principal stress at the osteotomy surface was at Z-axis. The direction was opposite to the Z-axis, and was the same as the total stress, which was a compressive stress. The shear force was the largest on the XY plane, and the maximum stress was at the dorsal inner side (A2) of the proximal osteotomy surface, which was 0.058 MPa. (3) The major displacements of the distal and proximal ends of the first patella osteotomy were on the X-axis, and the displacements were on the medial condyle (D1) of the osteotomy surface, i.e., -1.002 mm and medial condyle (A2), and 0.621 mm, respectively. (4) The results confirm that the external fixation of "8" bandage can maintain the stability of the osteotomy end after minimally invasive treatment of hallux valgus, and is conducive to the healing of the osteotomy end.

Analysis of Factors Influencing Overall Survival of MDS Patients Transplanted with HSCs / 中国实验血液学杂志

OBJECTIVE@#To analyze the effect of clinical features, routine laboratory examination and related gene mutation on the OS of patients with myelodysplastic syndrome (MDS) after hematopoietic stem cell transplantation (HSCT).@*METHODS@#121 patients diagnosed as MDS and underwent hematopoietic stem cell transplantation in the First Affiliated Hospital of Soochow University from October 2013 to August 2018 were selected. Basic information of the patients was collected, and blood cells, bone marrow blasts at initial diagnosis, chromosomal karyotypes and gene mutations of the patients were detected.The effect of different factors on overall survival (OS) was analyzed by statistical method.@*RESULTS@#Kaplan-Meier univariate analysis shows that OS was significanly different among different age groups. The 3-year OS rate of patients aged 0-29 years was (83.3±7.7) %, the 3-year OS rate in patients aged 30-49 years was (58.1±7.7 %), and the 3-year OS rate of patients aged 50-69 years was (31.0±22.6) %, which was statistically different (P＜0.05) between different groups. There were also significant differences in OS among patients with different transplantation types. 3-year OS rate: HLA-matched sibling HSCT＞unrelated HLA-matched HSCT＞haploidentical HSCT＞micro HSCT. The OS rate of patients with bone marrow blasts≥10% seems lower than blasts＜10%, but there was no statistical difference.The 3-year OS rate of patients with chromosomal karyotype complex abnormality was (47.7±11.5) %, and that of patients without complex abnormality was (80±4.2) % which was statistical difference (P＜0.05). Patients with DNMT3A, NRAS, TP53 and GATA2 mutations had shorter OS time compared with patients without mutation of these genes, which shows statistically significant (P＜0.05). COX multivariate analysis showed that age, chromosome karyotype, DNMT3A, TET2, GATA2 and NRAS were the independent factors influencing OS of patients after HSCT, with statistically significant difference.@*CONCLUSION@#age of patients, donor selection of HSCT, chromosome karyotype, DNMT3A, NRAS, TP53, GATA2 and TET2 gene mutations are all independent factors affecting the OS of patients after HSCT. Therefore, the assessment of the OS of MDS patients with transplantation requires comprehensive consideration.

Epigenetic Regulation and Epigenomic Landscape in Normal Physiological Hematopoiesis--Editorial / 中国实验血液学杂志

Abstract　　The physiological hematopoiesis depends on the programmed expression of a series of gene regulated by mechanisms at various levels. Currently, the epigenetic regulation has been considered as the most important mechanism during hematopoietic differentiation, resulting in a specific epigenomic landscape in the hematopoietic stem/progenitor cells. We try to concisely review the epigenetic mechanisms, including the genomic methylation, the histone modifications and the expression profiles of noncoding RNA, illustrating briefly the differentiation from the hematopoietic stem/progenitor cells to to the erythroid, myeloid and lymphoid cells.

Construction and expression of a recombinant protein with γ-aminobutyric acid type C receptor ρ2 subunits / 新乡医学院学报

Objective To construct prokaryotic expression vector of γ-aminobutyric acid type C receptor ρ2 (GABACR ρ2) gene,induce the expression of the recombination protein GABACR ρ2,and transfect the protein into SH-SY5Y cell line,achieve the transient expression of the recombination protein GABACR ρ2 in SH-SY5Y cell line.Methods The ρ2-Tat gene was inserted into plasmid pET30 to construct the recombinant plasmid pET-ρ2-GFP-Tat.The expression of GABACR ρ2 was detected by Western blot.The histidine-tagged recombination protein ρ2 was purified through immobilized Ni2+ absorption chromatographic column and the purity of GABACR ρ2 protein was detected by sodium dodecyl sulfate polyacrylamide gel elec trophoresis.The transduction and cellular localization of the GABAC R ρ2 was observed by fluorescence microscope.The SH-SY5Y cells were divided into normal group and hypoxia low glucose group.The cells in normal group were divided into normal control group which cultured in high glucose medium and without protein transfection and normal transfection group which cultured in high glucose medium and with protein transfection;the cells in hypoxia low glucose group were divided into treatment group which cultured under hypoxia low glucose condition and treatment transfection group which cultured under hypoxia low glucose and with protein transfection;the viability of SH-SY5Y cells in each group was evaluated by cell counting Kit-8.Results The recombinant plasmid pET30-ρ2-Tat was constructed successfully.The purified recombination protein ρ2-Tat successfully crossed the cytomembrane and transfected the SH-SY5Y cells.The viability of cells in normal control group,normal transfection group,treatment group and treatment transfection group was(100.0 ± 6.9)％,(89.3 ± 3.6)％,(51.4 ± 3.6)％and (66.1 ± 8.5) ％ respectively.There was no statistic difference in the cell viability between the normal control group and normal transfection group(P ＞0.05);the cell viability in treatment group was significantly lower than that in the normal control group (P ＜ 0.01);the cell viability in treatment transfection group was significantly higher than that in the treatment group (P ＜ 0.01).Conclusion The recombination protein ρ2 can through the membrane under the effect of Tat transduction peptide and can successfully establish a SH-SY5Y cell lines which transiently express recombinant protein ρ2,which provide a new research method for the study of ρ2 subunit function.

Effect of AML1-ETO Fusion Protein on Transcriptional Regulation of p14 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of transcriptional regulation of aberrant transcription factor AML1-ETO on p14.</p><p><b>METHODS</b>P14expression both in AML1-ETO-expressing cells or U937 nonexpressing cells and in leukemia cells of AML patients with or without t(8;21) was assessed by quantitative PCR. Methylation-specific polymerase chain reaction (MSP) was used to analyze the methylation status of p14promoter. The chromatin immunoprecipitation (ChIP)-based PCR was used to investigate the direct interaction between the AML1-ETO and p14promoter in AML1-ETO positive leukemia cell line. And the p14mRNA expression level was detected by qRT-PCR after treatment with 5-Aza.</p><p><b>RESULTS</b>AML1-ETO-expressing cell subclone displayed low level of p14mRNA in comparison with the non-transfected U937. In primary bone marrow cells of acute myeloid leukemia containing AML1-ETO, level of p14mRNA was markedly lower when compared with other acute myeloid leukemias lacking this translocation. P14gene promoter was non-methylated in control group and primary leukemia cells of AML patients without t(8;21) and was hyper-methylated in U937-A/E1-4 and primary leukemia cells of AML patients with t(8;21). The enriched regions in transfected cells were located within p14promoter. 5-Aza could increase the expression of p14.</p><p><b>CONCLUSION</b>P14is a possible target gene of AML1-ETO. The p14silencing induced by hyper-methlylation may be an important factor for occurrence and development of the Msubtype of acute myeloid leukemia.</p>

Autophagy Activity of CD34+ Cells in MDS Patients and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the autophagy activity of CD34+ cells in bone marrow of MDS patients and its clinical significance.</p><p><b>METHODS</b>The activity of autophagy in bone marrow CD34+ cells from 20 MDS patients, 20 non-malignant anemia patients and 5 AML patients admitted in our hospital from October 2012 to March 2014 was detected by flow cytometry (FCM).</p><p><b>RESULTS</b>The autophagy activity in low risk MDS patients and non-malignant anemia patients were both significantly higher than that in both high risk MDS and AML patients (P<0.05), and more interestingly, the autophagy activity in MDS negatively correlated with World Health Organization classification-based prognostic system (WPSS) score (r=-0.877) .</p><p><b>CONCLUSION</b>The autophagy activity CD34+ cells in the patients with MDS is higher than that in AML patients, and negatively correlated with WPSS scores, indicating that the decrease of autophagy activity maybe accelerate the genesis and development of MDS and relate with the prognosis of MDS patients.</p>

Differentiation of K562 Cells Induced by Pulsatilla Saponin A into Erythroid Lineage / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the differentiation-inducing potentiality of Pulsatilla saponin A on K562 cells.</p><p><b>METHODS</b>Pulsatilla saponin A of different concentrations was used to treat K562 cells; the benzidine staining and the hemoglobinometry were applied to measure the change of hemoglobin content; the flow cytometry (FCM) was used to detect the expression of CD71 and GPA on K562 cells.</p><p><b>RESULTS</b>K562 cells treated with 4 µg/ml pulsatilla saponin A differentiated into the erythroid lineage. With the treatment of pulsatilla saponin A, the hemoglobin content in K562 cells increased significantly; CD71 and GPA expression on the K562 cell surface were up-regulated.</p><p><b>CONCLUSION</b>Pulsatilla saponin A can induce K562 cells to differentiate into erythroid lineage.</p>

Clinical and Laboratorial Characteristics of Primary Acute Myeloid leukemia with Philadelphia Chromosome and Inversion 16 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To summarize the clinical characteristics as well as diagnosis and treatment in 1 case of acute myeloid leukemia(AML) with coexpression of Ph and inv(16).</p><p><b>METHODS</b>A series of clinical tests, the cellular morphological, immunological, cytogenetic and molecular biological examinations of leukemia cells were performed.</p><p><b>RESULTS</b>The clinical characteristics of this patient were very common. The cellular morphology is similar to the AML with inv(16). The leukemia cells were stained positively for CD13, CD33, CD34, CD117 and HLA-DR. Karyotypic analysis showed a complex chromosome abnormality including inv(16) and Ph, and the FISH analysis showed that the percentage of rearrangement of CBFβ allele was over that of the BCR-ABL fusion signals. The obvious adverse events did not occur in this patient within 3 years.</p><p><b>CONCLUSION</b>Ph as secondary aberration of inv(16) rarely occures in primary AML cases, and so far there have not been the clear criteria of diagnosis and treatment. The cytogenetic and molecular biology could provide the basis for diagnosis. Moreover, autologous hematopoietic stem cell transplantation combined with imatinib probably is one of the effective treatment methods.</p>

Diagnostic Utility of Diffusion-weighted Magnetic Resonance Imaging in Differentiating Small Solid Renal Tumors (≤ 4 cm) at 3.0T Magnetic Resonance Imaging / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The aim of this study was to assess the performance of apparent diffusion coefficient (ADC) measurement obtained with diffusion-weighted magnetic resonance imaging (DW-MRI) to distinguish renal cell carcinomas (RCCs) from small benign solid renal tumors (≤ 4 cm).</p><p><b>METHODS</b>In this cross-sectional study, 49 consecutive patients with histopathologically confirmed small solid renal tumors, and seven healthy volunteers were imaged using nonenhanced MRI and DW-MRI. The ADC map was calculated using the b values of 0, 50, 400, and 600 s/mm 2 and values compared via the Kruskal-Wallis and Mann-Whitney tests. The utility of ADC for differentiating RCCs and benign lesions was assessed using a receiver operating characteristic curve. Multiple nonenhanced MRI features were analyzed by Logistic regression.</p><p><b>RESULTS</b>The tumors consisted of 33 cases of clear-cell RCCs (ccRCCs) and 16 cases of benign tumors, including 14 cases of minimal fat angiomyolipomas and 2 cases of oncocytomas. The ADCs showed significant differences among benign tumors ([0.90 ± 0.52] × 10-3 mm 2 /s), ccRCCs ([1.53 ± 0.31] × 10-3 mm 2 /s) and the normal renal parenchyma ([2.22 ± 0.12] × 10-3 mm 2 /s) (P < 0.001). Moreover, there was statistically significant difference between high and low-grade ccRCCs (P = 0.004). Using a cut-off ADC of 1.36 × 10-3 mm 2 /s, DW-MRI resulted in an area under the curve (AUC), sensitivity, and specificity equal to 0.839, 75.8%, and 87.5%, respectively. Nonenhanced MRI alone and the combination of imaging methods led to an AUC, sensitivity and specificity equal to 0.919, 93.9%, and 81.2%, 0.998, 97%, and 100%, respectively. The Logistic regression showed that the location of the center of the tumor (inside the contour of the kidney) and appearance of stiff blood vessel were significantly helpful for diagnosing ccRCCs.</p><p><b>CONCLUSIONS</b>DW-MRI has potential in distinguishing ccRCCs from benign lesions in human small solid renal tumors (≤ 4 cm), and in increasing the accuracy for diagnosing ccRCCs when combined with nonenhanced MRI.</p>

Simultaneous determination six components in oolong tea by HPLC / 中国药学杂志

OBJECTIVE: To develop an HPLC method for simultaneous determination five eatechins, including (+)-catechin, epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate and (-)-epigallocatechin-3-gallate, as well as caffeine in Oolong tea. METHODS: The HPLC analysis was carried out on a Grace Allitima C18 column (4.6 mm × 250 mm, 5 μm), with a gradient elution acetonitrile-0.2% acetic acid at the flow rate 1.0 mL · min-1. The detective wave length was set at 280 nm, and the column temperature was set at 35℃. RESULTS: The calibration curve was linear within the range 5.95-952 μg · mL-1 for catechin, 6.36-1018 μg · mL-1 for epicatechin, 6.55-1048 μg · mL-1 for epigallocatechin, 7.80-1248) μg · mL-1 for epigallocatechin gallate, 10.83-1732 μg · mL-1 for epigallocatechin-3-gallate, and 13.38-2 140 μg · mL-1 for caffeine, respectively. The average recoveries for six determined marker compounds were 99.12%-102.12%. CONCLUSION: This method is simple, accurate, and practical for quality control Oolong tea. There are some differences in the contents the six marker compounds in the samples from different producing areas and different collection periods.

Frequently ABL kinase domain G:C→A:T mutation and uracil DNA glycosylase abnormal expression in TKI-resistant acute lymphoblastic leukemia of Chinese population / 中国实验血液学杂志

Most Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL) patients often show rapid recurrence and development of ABL kinase domain (KD) mutation after tyrosine kinase inhibitor (TKI) treatment. To further investigate the mechanism of Ph(+) ALL fast relapse after TKI treatment, ABL KD mutation in 35 Chinese Ph(+) ALL with TKI resistance was detected by direct sequencing. The results showed that 77.1% (27/35) Ph(+) ALL patients with TKI resistance had ABL KD mutation and 55.6% (15/27) Ph(+) ALL patients with ABL KD mutation had T315I. Interestingly, 77.8% (21/27) Ph(+)ALL showed ABL mutation G: C→A:T, including T315I, E255K and E459K. Furthermore, all the Ph(+) ALL patients with two or more ABL KD mutations collaborated with complex chromosome abnormality and all the TKI-resistant Ph(+) ALL patients, whose karyotype progressed from simple t (9;22) into complex, developed ABL KD mutation. Moreover, the expression level of uracil-DNA glycosylase UNG2, which inhibits G:C→A:T transition in genomic DNA, decreased in Ph(+) ALL with TKI-resistance compared to that in newly diagnosis Ph(+) ALL. It is concluded that there is a high frequent ABL KD G:C→A:T mutation and a high genomic instability in Chinese TKI-resistant Ph(+) ALL. In addition, the decreased UNG2 expression in TKI-resistant Ph(+) ALL probably contributes to their high rate of ABL KD G:C→A:T mutation.

Preliminary analyses of lymphocyte differentially expressed proteins in Huaihua Dong peoples with hypertension / 中国医师杂志

Objective To explore lymphocyte differentially expressed protein profile and its pathogenic significance in Huaihua Dong peoples with hypertension.Methods The lymphocyte protein profiles among 130 cases of blood specimen were detected and identified with matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF-MS) and Mascot search against protein database.Results The reproducible protein-maps of two-dimensional gel electrophoresis were obtained among three groups.Compared with Dong normal peoples,the protein disulfide isomerase-related protein 5 and heat shock protein 27 showed higher expressions in the Dong peoples with hypertension (P ＜ 0.01),but chain A,structure of haemoglobin in the deoxy quaternary state with ligand bound at the alpha haems is no statistical significance(P ＞0.05).Those three proteins were upregulated in Han peoples with hypertension (P ＜0.01).Conclusions The up-regulation of the protein disulfide isomerase-related protein 5 and heat shock protein 27 in the Dong and Han peoples with hypertension suggest that those two proteins might be associated with the occurrence and development of hypertension.

The different characteristics of ABL kinase domain mutation in the Chinese Han nationality imatinib resistant Philadelphia chromosome-positive acute lymphoblastic leukemia and chronic myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify the distribution and differentiation of ABL kinase domain mutation in the Chinese Han nationality imatinib resistant chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL).</p><p><b>METHODS</b>Bone marrow or peripheral blood samples of 112 imatinib resistant CML patients and 21 Ph(+)ALL patients were obtained from the first affiliated hospital of Soochow university according to local law. Total RNA was extracted from the mononuclear cells using a TRIzol reagent. ABL kinase domain (KD) mutation was detected by direct sequencing.</p><p><b>RESULTS</b>Of the 112 imatinib resistant CML patients, 54.46%(61 cases) had ABL KD mutation. Twenty-three mutants were identified in 20 amino acid sites and 23.21% (26 cases) ABL KD mutations were in P-loop region. ABL KD mutations were also detected in 71.43% (15 cases) imatinib resistant Ph(+)ALL patients, with 10 mutations in 8 amino acid sites. The most frequent mutation was T315I (28.57%), followed by E255K/V (19.05%) and Y253F/H (14.29%). The frequency of T315I was much higher in imatinib resistant Ph(+) ALL than that in imatinib resistant CML (P = 0.001). Ph(+)ALL with additional chromosomal aberrations also had a higher rate of ABL KD mutation than that of CML (P = 0.010). Ph(+)ALL gained ABL KD mutation faster than CML (P < 0.010).</p><p><b>CONCLUSION</b>Chinese imatinib resistant CML and Ph(+)ALL patients had different characteristics in ABL KD mutation. The rate of ABL KD mutation in Ph(+)ALL with additional chromosomal aberrations was much higher than that of CML with additional chromosomal aberrations.</p>

The influence of CD44 on the adhesive, migratory and infiltrative abilities of leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of CD44 in leukemia cell lines and its role in adhesion, migration and infiltration of leukemia cells.</p><p><b>METHODS</b>The expression levels of CD44 in four leukemia cell lines SHI-1, THP-1, NB4 and K562 were assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot when they were in logarithmic phase. And these cell lines were divided into control group (treated with same species and isotype IgG) and experimental group (treated with anti-CD44 mono-clonal antibody). The assays of cell-cell adhesion to endothelial cells line ECV304, migration through the artificial matrix membrane and infiltration through the Matrigel were performed.</p><p><b>RESULTS</b>The relative expression ratios of CD44 to GAPDH in SHI-1, THP-1, NB4 cells were 0.0731 ± 0.0072, 0.0827 ± 0.0151 and 0.1473 ± 0.0365, respectively, which were significantly higher than that in K562 cells (0.0002 ± 0.0000, P < 0.01). Cell-cell adhesion assay showed that the adhesion rates of SHI-1, THP-1 and NB4 cells in the experimental group decreased to 72.78%, 64.09% and 57.42%, respectively, and were lower than those of the control groups, while that of K562 cells in the experimental group was 106.16%. Migration assay showed that the transmembrane rates of SHI-1,THP-1 and NB4 cells were 55%, 29% and 25% in the control group, respectively, and decreased to 32%, 18% and 12% in the experimental group, respectively, while those of K562 cells in both control group and experimental group remained 2%. The infiltration rates of SHI-1, THP-1 and NB4 cells decreased from 24%, 15% and 13% in the control group to 12%, 8% and 4% in the experimental group, respectively, while K562 cells in both groups could not pass through the Matrigel.</p><p><b>CONCLUSION</b>CD44 antigen might play an important role in the adhesion, migration and infiltration of leukemia cells and be involved in the extra-medullary infiltration of leukemia cells.</p>

Transfection of HL-60 cells by Venus lentiviral vector / 中国实验血液学杂志

In order to study the potential of Venus, lentiviral vector, applied to acute myeloid leukemia, the recombinant vector Venus-C3aR was transfected into 293T packing cells by DNA-calcium phosphate coprecipitation. All virus stocks were collected and transfected into HL-60, the GFP expression in HL-60 cells was measured by flow cytometry. The expression level of C3aR1 in transfected HL-60 cells was identified by RT-PCR and flow cytometry. The lentiviral toxicity on HL-60 was measured by using CCK-8 method and the ability of cell differentiation was observed. The results indicated that the transfection efficacy of lentiviral vector on HL-60 cells was more than 95%, which meets the needs for further study. C3aR1 expression on HL-60 cells increased after being transfected with recombinant lentiviral vector. Before and after transfection, the proliferation and differentiation of cells were not changed much. It is concluded that the lentiviral vector showed a high efficacy to transfect AML cells and can be integrated in genome of HL-60 cells to realize the stable expression of interest gene. Meanwhile, lentiviral vector can not affect HL-60 cell ability to proliferate and differentiate.

Study of the clonal origin and development of MDS by FISH analysis of dysplasia cells in bone marrow of patients with MDS / 中国实验血液学杂志

This study was purpose to explore whether the dysplasia of myelodysplastic syndromes (MDS) is unspecific feature or results of the abnormal clone, and to provide the evaluation of abnormal clone changes in bone marrow cells of MDS patients. The dysplasia cells in bone marrow smears was analyzed by morphologic observation, the clonal origin and development in 16 cases of MDS with abnormality of chromosome karyotypes were investigated by FISH combined with morphologic observation. The results found that both the dysplastic and nondysplastic bone cells displayed abnormal clones in the erythroid and granulocytic cells. The dysplastic bone marrow cells displayed more abnormal clones than the nondysplastic bone marrow cells in most of the patients, and the abnormal clones displayed more dysplastic cells than the normal clones. Most of the dysplastic and nondysplastic megakaryocytes were derived from abnormal clones. The abnormal clone showed a decreasing trend from the primitive stage to the terminal stage of cell differentiation. It is concluded that there is a correlation between the dysplastic cells and the abnormal clones in MDS, but the dysplasia of bone marrow cells is not a specific feature. The abnormal clones can differentiate into mature granulocytes and erythrocytes, and can be in coexistence with cells originated from the normal clones.

Biocompatibility of polyethylene imine (PEI)-coated magnetic Fe₃O₄ nanoparticles in SHI-1 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility of magnetic resonance cell imaging technology by using polyethylene imine (PEI)-coated magnetic nanoparticles of Fe₄O₄ (PEI-Fe₄O₄-MNPs) to track cell biology behavior.</p><p><b>METHODS</b>Endocytic PEI-Fe₄O₄-MNPs in SHI-1 cells were observed by transmission electron microscopy (TEM) . Iron contents of nano-labeled cells were analyzed by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) and Prussian blue staining. The proliferation ability of labeled cells was detected by cell counting kit-8 (CCK-8) assay; the differentiation and colony-forming abilities were also observed. SHI-1 cells without endocytosing PEI-Fe₄O₄-MNPs were used as control.</p><p><b>RESULTS</b>Our data showed that PEI-Fe₄O₄-MNPs could label SHI-1 cells. The labeling efficiency depended on the nanoparticles' concentration and the duration of cells treating. Inhibition rates of SHI-1cells labeled by 60-100 μg Fe/ml PEI-Fe₄O₄-MNPs were much higher than of 5-50 μg Fe/ml ones following treating by 5-100 μg Fe/ml PEI-Fe₄O₄-MNPs for 48 hrs. The expressions of CD11b and CD14 were (78.4±18.5)% and (18.7±2.9)% in control vs (83.3±14.2)% and (20.4±2.1)% in cells fractions treated by 30 μg Fe/ml PEI-Fe₄O₄-MNPs. Clony-forming rates of SHI-1 cells labeled by 0, 20 , 50 μg Fe/ml PEI-Fe₄O₄-MNPs were (25.20±7.22)%, (25.93±13.15)%, (23.37±9.33)%, respectively. Differentiation and colony-forming potentials of labeled cells were similar with control in the certain range of PEI-Fe₄O₄-MNPs concentration.</p><p><b>CONCLUSION</b>SHI-1 cells were efficiently labeled by PEI-Fe₄O₄-MNPs with well biocompatibilities in proper range of concentration, the latter could be coupled with magnetic resonance imaging (MRI) to track cells in vivo.</p>