AIMTo explore whether the oligonucleotide uptake in hematological tumor cells is related to cellular species and proliferation.

METHODSIntracellular mean fluorescence intensity was measured by flow cytometry.

RESULTSAfter treatment with FITC-labeled G3139 at the concentration of 0.60 mumol.L-1 for 4 h, the G3139 uptake into peripheral blood mononuclear cell and bone marrow mononuclear cell in hematological tumor patients was significantly higher than that in normal control. There was different uptake of G3139 among the malignant hematological tumor cell strains, and the uptake in cells derived from monocyte, B lymphocyte and myeloid cell was much higher than that in cells derived from T lymphocyte. After treatment with all-trans retinoic acid (ATRA), HL60 cell proliferation was markedly inhibited and the uptake of G3139 decreased significantly.

CONCLUSIONHematological tumor cells were capable of taking up oligonucleotide, and the oligonucleotide uptake in hematological tumor cells is related to its cellular species and its activation.

Biological Transport ; Cell Division ; physiology ; Genes, bcl-2 ; genetics ; HL-60 Cells ; Humans ; Leukemia ; metabolism ; pathology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Leukocytes, Mononuclear ; metabolism ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; Oligonucleotides, Antisense ; metabolism ; Thionucleotides ; metabolism ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

The objective was to explore the feasibility of differentiation of human umbilical cord blood mononuclear cells into endothelial cells induced by cytokines in vitro and to study the possibility of using cord blood stem cells in ischemic diseases therapy. The cells were isolated from umbilical cord blood by using lymphocyte separation solution, and committedly differentiated by using VEGF, bFGF and IGF-I in a liquid culture system. The results showed that the combination of cytokines produced a large number of caudated adherent cells and flow cytometric analysis revealed endothelial marker vWF expressed in about 80% cells, and the endothelial -specific Weibel-Palade body was detected in the cytoplasm by electronic microscope. It is concluded that human umbilical cord blood mononuclear cells may be induced to differentiate into endothelial cells induced by VEGF, bFGF and IGF-I. Human umbilical cord blood MNC may be an ideal source of adult stem cells for the treatment of the ischemic disease.
Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; ultrastructure ; Fetal Blood ; cytology ; Fibroblast Growth Factor 2 ; pharmacology ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Leukocytes, Mononuclear ; cytology ; Microscopy, Electron ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo investigate the outcomes after 2 methods of laparoscopic gastric bypass surgery for patients with type 2 diabetes mellitus(T2DM).

METHODSFrom December 2009 to June 2011, 21 patients with T2DM underwent laparoscopic gastric bypass surgery, including laparoscopic Roux-en-Y gastric bypass (LRYGB, n=11), and laparoscopic mini-gastric bypass (LMGB, n=10). Clinical data were analyzed retrospectively.

RESULTSThe clinical complete remission rate of T2DM was 64%(7/11) in LRYGB group, and 60%(6/10) in LMGB group. The clinical partial remission rate of T2DM was 36%(4/11) in LRYGB group, and 40%(4/10) in the LMGB group. There was no significant difference between the two groups(both P>0.05). The levels of BMI, waist circumference, HOMA-IR and HbA1c within the postoperative 6 months were improved in each group (all P<0.05), but there was no significant difference between the two groups(all P>0.05). There were no conversion or perioperative deaths in both groups. Compared to LMGB, the LRYGB group had longer operative time[(147.0±35.9) min vs. (110.5±39.7) min, P=0.038] and postoperative hospital stay [(8.9±2.3) d vs. (7.1±1.4) d, P=0.046). One patient suffered from ileus in LRYGB group, one patient suffered from reflux esophagitis and one suffered chronic diarrhea in LMGB group. The incidence of postoperative complication was similar between the two groups(P>0.05).

CONCLUSIONLRYGB and LMGB may result in satisfactory and safe effects for the treatment of T2DM, while the LMGB is simpler and associates with quicker recovery.

Adult ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Gastric Bypass ; methods ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo obtain recombinant human interleukin 12 by genetic engineering, and to explore possibility of its clinical application in treatment of tumor and chronic hepatitis.

METHODSBicistronic expression vector P35-IRES-P40 was constructed for the simultaneous translation of IL-12 p35 and p40 cDNA subunit through internal ribosomal entry sites (IRES). pCI-dhfr-P35-IRES-P40 vector was constructed for expression in CHO-DHFR- cells. Positively cloned cells were screened by means of ELISA. Pools of clones with increased expression of IL-12 could be generated by selection in methotrexate. To determine the biological activities of rhIL-12, PHA-activated lymphoblasts proliferation assay and IFN-gamma induction assay were used in this study.

RESULTSGenetically engineered cells expressing hIl-12 were obtained and all the cell lines showed the stabile expression of rhIL-12 in high efficiency and good growth properties.

CONCLUSIONrhIL-12 have good biological activities, it can stimulate activation and proliferation of T cells and induce production of IFN-gamma.

Animals ; Blotting, Western ; CHO Cells ; Cell Proliferation ; drug effects ; Cricetinae ; Cricetulus ; Dose-Response Relationship, Drug ; Genetic Vectors ; genetics ; Humans ; Interleukin-12 ; biosynthesis ; genetics ; pharmacology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; metabolism ; pharmacology ; T-Lymphocytes ; cytology ; drug effects ; Transfection

This study was purposed to construct a vector containing human suppressor gene p53 and p16, and to investigate their expression and effect on K562 and HL-60 cells. pBudCE4.1-53-16 is a vector designed for simultaneous expression of human suppressor gene p53 and p16 in mammalian cell line. After transfection into K562 cells with lipofectamine(TM) 2000, the expression of p53 and p16 genes was detected by Western blot and immunocytochemical method. The growth curve, apoptosis, cell cycle were assayed by CCK-8 and flow cytometry. The results showed that the recombinant plasmid pBudCE4.1-53-16 was constructed successfully and were verified by PCR and restriction analysis. The expression of P53 and P16 protein could be detected after transfection into leukemia cells (K562 and HL-60) for 48 hours. As compared with control group, the cell proliferation in experimental group was inhibited, the cells were arrested in G0 phase and apoptotic cells increased (p<0.001). It is concluded that the recombinant plasmid pBudCE4.1-53-16 has been established. p16 and p53 in the recombinant plasmid pBudCE4.1-53-16 synchronously express in leukemic cells after transfection in vitro for 2 days and results in reduced proliferation, G0 arrest and apoptosis increase.
Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Gene Expression ; Genes, p53 ; Genetic Vectors ; HL-60 Cells ; Humans ; K562 Cells ; Plasmids ; Transfection

OBJECTIVETo evaluate the clinical outcome of reconstruction of maxillary defects with vascularized iliac crest flap and simultaneous osseointegrated implant embedding.

METHODSDuring September to October 2003, two patients with maxillary defects from tumor resection underwent microsurgical reconstruction. The free iliac osteomuscular flap transferring and simultaneous osseointegrated implant embedding were performed to repair the defects. Three months after the reconstructive surgery, an abutment operation was preformed and denture was applied in both cases.

RESULTSThe flaps survived well. Postoperative follow-up for 8 to 9 months showed that the patients obtained good zygomaxillary appearance, normal occlusion, and satisfactory pronunciation, without oronasal fistula or other serious complications.

CONCLUSIONSThe free iliac crest osteomuscular flap with simultaneous osseointegrated implant embedding is an ideal, effective and cosmetically acceptable method for maxilla reconstruction.

Adult ; Bone Transplantation ; methods ; Female ; Humans ; Ilium ; transplantation ; Male ; Maxilla ; surgery ; Middle Aged ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo summarize the surgical technique and perioperative management of laparoscopic sleeve gastrectomy (LSG).

METHODSA total of 57 morbid obesity patients undergoing LSG surgery from May 2010 to December 2012 were enrolled in the study, whose clinical data in perioperative period were analyzed retrospectively. These patients had more than 1 year of follow-up. All the patients received preoperative preparation and postoperative management, and postoperative excess weight loss(EWL%) and improvement of preoperative complications was evaluated.

RESULTSAll the cases completed the operation under laparoscopy, except 1 case because of the abdominal extensive adhesion. The average operation time was(102.0±15.2) min and the mean intraoperative blood loss (132.3±45.6) ml. Of 2 postoperative hemorrhage patients, 1 case received conservative treatment, and another one underwent laparoscopic exploration. The EWL% at 3 months, 6 months and 1 year after procedure was (54.9±13.8)%, (79.0±23.6)% and (106.9±25.1)% respectively. The preoperative complications were improved in some degree. There were no operative death, and anastomotic leak, anastomotic stenosis, or surgical site infection occurred.

CONCLUSIONLSG is a safe and effective surgical technique, whose safety and efficacy may be increased by improving the perioperative management.

Gastrectomy ; methods ; Humans ; Laparoscopy ; Obesity, Morbid ; Retrospective Studies ; Weight Loss

OBJECTIVETo compare two fluorochrome staining methods for the assessment of sperm quality.

METHODSWashed sperm cells were incubated in 0, 0.15, or 15 micromol/L camptothecin (CAM), or 0.37 or 3.7 mmol/L genistein (GEN) at 37 degrees C for 4 hours. The sperm cells were analyzed for cycle-independent apoptosis and necrosis by single-stain compared with dual-stain fluorescence microscopy to contrast the relative effectiveness of these two approaches.

RESULTSThe single-stain procedure could not detect the sperm viability differences. In contrast, the dual-stain procedure identified a dosage-dependent decrease in the viability and increased necrozoospermia after topoisomerase inhibitor CAM and GEN treatments. Apoptosis was 2-fold higher with topoisomerase inhibitor treatment.

CONCLUSIONThe two topoisomerase inhibitors were associated with increased apoptosis and dosage-dependent necrosis. The data suggested that the dual-stain combination Hoechst 33342/PI was more sensitive than the single Hoechst 33342 stain analysis and permitted quantitative analysis of the apoptosis and necrosis in sperm.

Apoptosis ; drug effects ; Benzimidazoles ; Camptothecin ; pharmacology ; Cell Survival ; drug effects ; DNA ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Fluorescent Dyes ; Genistein ; pharmacology ; Humans ; Male ; Propidium ; Sensitivity and Specificity ; Spermatozoa ; drug effects ; physiology ; Staining and Labeling ; methods

The tumor suppressor gene p53 and p16, both of which play an important role in inhibition of tumorigenesis, are homozygously deleted in human myeloid leukemia cell line K562. To explore the inhibition of K562 cell proliferation by wild type p16 and p53 genes, both p16 and p53 genes were co-transfected into K562 cells mediated by liposome. The expression of the two genes was measured by immunocytochemical method, the cell cycle was analysed by flow cytometry, and the number of recovered viable cells was assessed after transfection. After co-transfection, the p53 and p16 positive cells were 23% and 28%, respectively. The results showed that co-transfection of p16 and p53 genes significantly inhibits cell proliferation comparing with transfection either by p16 gene or by p53 gene (P < 0.05). Expression of p16 and p53 proteins increased the cell number in G(1) phase but decreased the cell number in S phase. It is concluded that co-transfection of p16 and p53 genes has a stronger growth-inhibitory effect on K562 cell growth than that of transfection only by p16 gene or by p53 gene, may be a pathway for gene therapy in leukemia.
Cell Division ; Genes, p16 ; physiology ; Genes, p53 ; physiology ; Humans ; K562 Cells ; Plasmids ; Transfection

Chronic Disease ; Coronary Occlusion/surgery* ; Humans ; Treatment Outcome ; Ultrasonography, Interventional