The purpose of this study was to explore the expressions of midkine (MK) and vascular endothelial growth factor (VEGF) in multiple myeloma (MM), and to evaluate their relation with angiogenesis and prognosis. The expression levels of MK and VEGF in bone marrow mononuclear cells of 31 MM patients in different stages and 20 controls were detected by real-time fluorescent quantitative RT-PCR. The results showed that the MM patients had significantly higher MK and VEGF expression level than control group (p < 0.05, p < 0.01), and there was a linear relationship between MK and VEGF (r = 0.692, p < 0.01); The expression levels of MK and VEGF in stage III was significantly higher than those in stage I and stage II (p < 0.05, p < 0.01), but there was no difference between stage I and stage II (p > 0.05); MK and VEGF levels were significantly decreased in MM patients after treatment than those before treatment (p < 0.05, p < 0.01). It is concluded that the high expression of MK and VEGF is correlated with angiogenesis and prognosis of MM, and there is synergistic effect between MK and VEGF. It is supposed that the monitoring MK and VEGF expression levels may contribute to guide the treatment and estimate prognosis for MM.
Adolescent ; Adult ; Aged ; Bone Marrow ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; metabolism ; pathology ; Neoplasm Staging ; Nerve Growth Factors ; metabolism ; Prognosis ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

This study was purposed to investigate the significance of Id4 gene methylation in monitoring the efficacy of allo-PBSCT for treatment of acute leukemias. MS-PCR method was used to detect Id4 gene methylation in bone marrow samples from 29 patients with acute leukemia before and at 1, 3, 6 and 12 months after allo-PBSCT. The results showed that the Id4 gene was methylated in 18 patients before allo-PBSCT, out of which Id4 gene methylation in 8 patients could be detected sustainedly after allo-PBSCT, whereas among remaining 11 patients with Id4 gene unmethylation before PBSCT, the Id4 gene of only one case was found to be methylated after PBSCT. Out of 9 patients with Id4 gene methylation after allo-PBSCT, 4 had relapse during the follow-up. 20 patients with Id4 gene unmethylation after allo-PBSCT were in continuously complete remission status. Id4 gene methylation was found more frequently between 6 months and 1 year after allo-PBSCT. It is concluded that detecting Id4 gene methylation is important for the AL patients who underwent allo-PBSCT. Choosing the patients with Id4 gene unmethylaiton to receive allo-PBSCT may help to reduce relapse rate. After allo-PBSCT, Id4 gene methylation status can be regarded as an indicator for predicting prognosis of acute leukemias.
Adult ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia ; genetics ; surgery ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Young Adult

OBJECTIVESTo identify and clone human genes transactivated by HCV F protein by constructing a cDNA subtractive library using the suppression subtractive hybridization technique.

METHODSSuppression subtractive hybridization (SSH) and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV F protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1 (-)-F or with pcDNA3.1(-) empty vector as a control, and SSH method was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small sized cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 or adaptor 2. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5 alpha. The cDNA was sequenced and analyzed in GenBank with blast search after PCR.

RESULTSThe subtractive library of genes transactivated by HCV F protein was constructed successfully. The amplified library contains 71 positive clones. Colony PCR shows that 56 clones contain 200-1000 bp inserts. Sequence analysis was performed on 28 clones randomly, and the full length sequences were obtained with using the bioinformatics method. Altogether 19 coding sequences were obtained, consisting of 17 known and 2 unknown.

CONCLUSIONSThe obtained sequences may be target genes transactivated by HCV F protein, and some gene coding proteins are those involved in cell cycle regulation, metabolism, and cell apoptosis.

Cloning, Molecular ; Hepacivirus ; genetics ; Humans ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Transcriptional Activation ; Viral Core Proteins ; biosynthesis ; genetics

OBJECTIVEThe traditional gracilis musculocutaneous flap is supplied by a branch of deep femoral artery, which enters the muscle in between the upper and middle third of it. So the flap barely reaches the pelvis and perineum region for reconstruction. By exploring the blood supply pattern we tried to rotate the flap Upon at the higher point starting at the obturator foramen in order to let it cover a bigger area.

METHODSanatomical reviewing of the blood supply of the gracilis branches of obturator, medial femoral circumflex and deep femoral arteries. Based on this a new type of longitudinal gracilis musculocutaneous flap supported only by the obturator artery was designed to reach the pelvis, female genitalia, pubic symphysis, inguinal area easily.

RESULTSThe new kind of flap has been applied to 9 patients for deformity repairing and tissue replacement in the pelvic and perineal area. All the flaps survived and achieved satisfactory result with 3 months to 3 years' follow up.

CONCLUSIONSLongitudinal gracilis musculocutaneous flaps supplied by the obturator artery can be used as regular musculocutaneous flap clinically.

Female ; Femoral Artery ; surgery ; Humans ; Muscle, Skeletal ; blood supply ; transplantation ; Surgical Flaps ; blood supply

OBJECTIVETo separate the cell subpopulation with high tumorigenic ability and study the biological characteristics of this subpopulation in laryngeal carcinoma cells.

METHODSHuman laryngeal carcinoma cells were obtained by primary tissue culture technique. CD44 and CD133 molecules were used as markers to isolate the CD44(+), CD133(+), CD44(+)CD133(+) and CD44(+)CD133(-) cell subpopulations from the laryngeal carcinoma cells by flow cytometry. A nude mouse tumor xenograft model was developed for the study of the tumorigenic effects of the different cell populations. 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells were injected into the left flank of the mice, respectively. The mice were observed for palpable tumor formation and were sacrificed at 4 weeks later to assess the tumor formation rate, tumor volume and tumor weight. Boyden chamber migration assay was used to determine the migration ability and immunochemistry was used to detect the expression of stem cell antigen SCA-1 and beta1-integrin. Semi-quantities RT-PCR and Western blot analysis were performed to detect the expression level of Bmi-1 in the different cell subpopulations.

RESULTSThe growth of subcutaneous tumors in nude mice showed that a tumor can be generated with 1 x 10(3) CD44(+)CD133(+) cells. When the same dose of 1 x 10(6) CD44(+)CD133(+) cells was injected into the mice, both the average weight and volume of the tumors were significantly higher than those generated from other cell subpopulations. Boyden chamber migration assay showed that the invasion ability of CD44(+)CD133(+) cells was significantly higher than that of other cell subsets. The results of immunochemical analysis showed an abundant expression of stem cell antigen SCA-1 and beta1-integrin in the CD44(+)CD133(+) cells. Semi-quantitative RT-PCR and Western blot analysis provided strong evidence that the Bmi-1 expression in CD44(+)CD133(+) and CD133(+) cells was very significantly higher than that in CD44(+), CD44(+)CD133(-) and control cells (P < 0.01).

CONCLUSIONOur findings demonstrate that CD44(+)CD133(+) subset cells in laryngeal carcinoma posses some biological characteristics of tumor stem cells, which may be the original cells of laryngeal carcinoma and may become a new target of tumor therapy.

AC133 Antigen ; Animals ; Antigens, CD ; analysis ; Antigens, Ly ; metabolism ; Cell Adhesion ; Gene Expression Regulation, Neoplastic ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Integrin beta1 ; metabolism ; Laryngeal Neoplasms ; immunology ; metabolism ; pathology ; Male ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; analysis ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Tumor Burden ; Tumor Cells, Cultured

OBJECTIVETo investigate the changes in serum neuron-specific enolase (NSE) and serum ferritin (SF) in patients with pneumoconiosis and their relationship with the onset of pneumoconiosis.

METHODSThe serum NSE and SF levels in the peripheral blood of patients with pneumoconiosis were measured by electrochemical fluorescence immunoassay.

RESULTSThe patients with first-stage pneumoconiosis and second-stage pneumoconiosis had significantly higher serum NSE and SF levels than the control group (23.0264±14.0410 and 44.9776±26.5208 ng/ml vs 8.1480±3.7512 ng/ml, P < 0.05; 267.2515±186.5809 and 579.1371±433.9326 ng/ml vs 120.8613±74.2809 ng/ml, P < 0.05), and the patients with second-stage pneumoconiosis had significantly higher serum NSE and SF levels than those with first-stage pneumoconiosis (P < 0.05). After treatment, the serum NSE level decreased significantly in the patients with pneumoconiosis (21.1675±17.5942 ng/ml vs 33.4490±21.6948 ng/ml, P < 0.05), but it was still significantly higher than that in the control group (P < 0.05). The treatment did not produce significant changes in SF level among these patients (P > 0.05).

CONCLUSIONPatients with pneumoconiosis have elevated serum NSE and SF levels, which may be related to the onset and progression of this disease.

Adult ; Ferritins ; blood ; Humans ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; blood ; Pneumoconiosis ; blood ; Young Adult

OBJECTIVETo analyse the WT1 expression and its DNA methylation status of its promoter domain.

METHODThe expression of WT1 gene and its DNA methylation status were assayed in leukemia cell lines and normal peripheral blood mononuclear cells (PBMNC) by RT-PCR and MS-PCR.

RESULTSWT1 was overexpressed in HL60, K562 and KG1 leukemia cell lines, but not in U937 and PBMNC. Methylation of WT1 promoter was not observed in HL60 cells.

CONCLUSIONDNA methylation of WT1 gene promotor did not inhibit its expression. Other mechanisms may appear to regulate the WT1 expression.

Cell Line, Tumor ; DNA Methylation ; Genes, Wilms Tumor ; Humans ; Leukemia ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic

In order to study the epidemiological characteristics of cytomegalovirus (CMV) infection in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients by means of plasma real time quantitative polymerase chain reaction (RQ-PCR), 141 adult patients undergoing allo-HSCT between January 2008 and June 2010 were serially monitored by RQ-PCR for detecting CMV and guiding the preemptive therapy followed up to 180 days post-HSCT. The results showed that the incidence of CMV infection and CMV pneumonia was 81.5% and 2.9% respectively, which mainly occurred within 2 months post-HSCT. Single-therapy with ganciclovir (GCV) for 63 patients or foscarnet 6 patients was performed for preemptive therapy. The total efficacy was 87.8%, and the response patterns were different. CMV infection was more frequent in female patients (P = 0.044), and those with aGVHD (P = 0.043), using ATG or basiliximab in conditioning regimens (P = 0.049), as well as earlier in patients using ATG or basiliximab or those with aGVHD (P = 0.007; P = 0.000). The aGVHD, maximum load, positive times of CMV-DNA detection and therapy duration all correlated with the efficacy (P < 0.05). It is concluded that the incidence of CMV infection is still high after HSCT. Plasma RQ-PCR assay for CMV-DNA shows a strong correlation with the clinical outcome of CMV infection, which is useful and suitable for management of CMV infection in HSCT.
Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Child ; Cytomegalovirus ; Cytomegalovirus Infections ; diagnosis ; drug therapy ; etiology ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Transplantation, Homologous ; Young Adult

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics

This study was purposed to investigate the changes of serum immunoglobulin (Ig) level during treatment of diffuse large B cell lymphoma by using rituximab (RTX) combined with CHOP. Total of 122 newly diagnosed patients with CD20(+) diffuse large B cell lymphoma from January 2004 to December 2009 were analyzed retrospectively. According to different treatment regimens, 122 patients were divided into 2 group: group treated with CHOP (n = 24) and group treated with R-CHOP (n = 98, out of which 16 patients with abnormal Ig levels before treatment). Ig levels of patients in 2 groups at different stages were recorded and analyzed after abandoning those patients with abnormal Ig levels before treatment. The results showed that after 6 cycles of treatment, among the total 82 patients with normal levels of serum immunoglobulin, the decreased levels of IgG, IgA and IgM by 20% of baseline value were found in 85.4% (70/82), 85.4% (70/82) and 87.8% (72/82) patients respectively, while levels of IgG, IgA and IgM < low limit of normal value were observed in 47.6% (39/82), 48.8% (40/82) and 52.4% (43/82) patients respectively. No obvious changes of IgG, IgA and IgM levels were found in 24 patients of CHOP group before and after treatment.It is concluded that hypogammaglobulinemia is a common complication in chemotherapy using RTX combined with CHOP, the decreased level of Ig is recovered to normal level about 1 year after stop of treatment, the decrease of Ig in some cases can last even for over 2 years.
Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Humans ; Immunoglobulins ; blood ; Lymphoma, Large B-Cell, Diffuse ; blood ; drug therapy ; immunology ; Prednisone ; therapeutic use ; Prognosis ; Retrospective Studies ; Rituximab ; Treatment Outcome ; Vincristine ; therapeutic use