Hematologic Diseases ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells

Adolescent ; Adult ; Blood Donors ; Child ; Child, Preschool ; Female ; Gastrointestinal Diseases ; drug therapy ; etiology ; Graft vs Host Disease ; drug therapy ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Hormones ; administration & dosage ; therapeutic use ; Humans ; Male ; Middle Aged ; Transplantation, Homologous ; Young Adult

This study was aimed to explore the factors impacting on effect of the recombinant human granulocyte colony-stimulating factor (rhG-CSF) in mobilizing and collecting peripheral blood stem/progenitor cells (PBSPC) from healthy donors, and to determine the optimal time for PBSPC harvest. A mobilization course in 431 healthy donors was retrospectively studied and the factors influencing the efficacy of mobilization were analyzed. The normal donors underwent leukapheresis for PBSPC collection in multicentres after mobilization with G-CSF administered. A variety of items analyzed included donor age, sex, weight, body mass index (BMI), daily G-CSF dose and schedule of G-CSF administration. The results showed that G-CSF was administered subcutaneously at median 5.7 microg/kg for mobilization for 3 - 5 days, The median number of peripheral blood mononuclear cells (PBMNC) count of per kg recipient weight was 9.57 x 10(8) and CD34(+) cells per kg recipient weight was 4.91 x 10(6) after a median of 1.7 leukapheresis. The side effects were mild and well tolerated. By univariate analysis, BMI, daily G-CSF dose and schedule of administration were significantly correlated with the yield of PBMNCs, CD34(+) cells. The best apheresis yields of PBMNCs and CD34(+) cells were achieved on day 5 after treatment with rhG-CSF. Because the narrow range and low dose of rhG-CSF administration, there were minor effects of rhG-CSF dose compared with schedule of administration. It is concluded that mobilization and leukapheresis are safe in healthy donors and that the low dose of rhG-CSF in 5-day administration are probably optimal for donor management.
Adult ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; Humans ; Leukapheresis ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Recombinant Proteins ; Tissue Donors ; Young Adult

Objective To examine the complement component 1 Q subcomponent-binding protein (C1QBP) gene expression in human resistance choriocarcinoma cell lines and its parental cell line JeG-3,and to investigate whether silence C 1QBP by small interference RNA could reverse the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.Methods Expression of C1QBP mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR and western blot,respectively.The difference of C 1QBP expression was compared between human resistance choriocarcinoma cell lines and its parental cell line JeG-3.Sub-cellular location was proved by confocal immunofluorescence microscopy.A lentiviral vector containing short hairpin RNA (shRNA) targeting C 1QBP was constructed and cotransfected with the packaging plasmid mixture into 293T cells by lipofectamine 2000.The human resistance choriocarcinoma cell lines were infected with the packaged lentivirus.Real-time fluorogenic quantitative PCR and western blot were used to validate whether the C 1QBP gene expression was silenced.The cell counting kit 8(CCK8)was used to determine the drug sensitivity.Results (1)The C1QBP mRNA expression levels among four human resistance choriocarcinoma cell lines[JeG-3/floxuridiuum (FUDR),JeG-3/methotrexate (MTX),JeG-3/etoposide (VP),JeG-3/dactinomycin (KSM)] were 2.520±0.680,1.770±0.230,1.940±0.090 and 1.740±0.350 folds compared to that in JeG-3 cells.The C1QBP protein was higher expression level in human resistance choriocarcinoma cell lines than that in JeG-3.The immunofluorescence methods and confocal analysis showed that C1QBP localized predominantly in the mitochondrial matrix.(2)The C1QBP mRNA expression in JeG-3/FUDR cells after infected with lentiviral vector were decreased by 93.1％ (P＜0.01).The protein expression of C 1QBP in JeG-3/FUDR cells after infected with lentiviral vector were almost completely suppressed.The resistance indexes of four human resistance choriocarcinoma cell lines(JeG-3/FUDR,JeG-3/MTX,JeG-3/VP,JeG-3/KSM) were respectively 86.3％,93.9％,92.8％ and 89.9％,which were decreased remarkably by knockdown the C 1QBP expression (P＜0.05).Conclusions C1QBP is overexpressed in human resistance choriocarcinoma cell lines compared with parental cell line JeG-3.Inhibition of C 1QBP by lentivirus-mediated small interference RNA could effectively reverses the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.

The study was purposed to investigate the effects of humanized recombined CD25 monoclonal antibody (rhCD25MAb) on activation and proliferation of T lymphocytes in vitro. Peripheral blood mononuclear cells (PBMNC) were incubated with phytohemagglutinin (PHA). Before or after T cell activation, the cells were cultured with or without rhCD25MAb or cyclosporine A (CsA) in vitro. After 72 hours incubation, the proliferation of lymphocytes was analyzed by MTT assay. The expression of CD3 and CD25 antigens on T lymphocytes were detected by flow cytometry. The levels of sIL-2R in the supernatants were determined with ELISA. The results showed that both rhCD25MAb and CsA could inhibit the proliferation of T lymphocytes significantly in concentration-dependent manner and CsA was more efficient than rhCD25MAb. Both rhCD25MAb and CsA could also decrease the levels of sIL-2R in the supernatant and inhibit the expression of CD25 antigen on T lymphocytes. The level of sIL-2R and the expression of CD25 on T lymphocytes decreases more profoundly in rhCD25MAb group. It is concluded that rhCD25MAb shows strong immunosuppressive activity both before and after T cell activation, suggesting that this agent may be useful in not only prophylaxis but also the treatment of acute graft-versus-host disease.
Antibodies, Monoclonal ; pharmacology ; CD3 Complex ; biosynthesis ; genetics ; Cell Proliferation ; Cells, Cultured ; Cyclosporine ; pharmacology ; Humans ; Immunosuppressive Agents ; pharmacology ; Interleukin-2 Receptor alpha Subunit ; biosynthesis ; genetics ; immunology ; Lymphocyte Activation ; immunology ; Receptors, Interleukin-2 ; analysis ; Recombinant Proteins ; immunology ; pharmacology ; T-Lymphocytes ; immunology

This study was aimed to investigate the effect of STI571, an inhibitor of tyrosine kinase, on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells as well as expression of anti-apoptotic protein Mcl-1, and to explore the possible role of Mcl-1 in apoptosis-inducing mechanism. K562 cell line was used to observe the effect of STI571 on CML cells. Proliferation and cytotoxicity were analyzed by MTT assay. The apoptotic cells were labelled with Annexin V-FITC and PI and then analyzed by flow cytometry. The expression of apoptotic-related proteins in K562 cells was determined by Western blot with specific antibodies. The results showed that STI571 significantly inhibited the proliferation and induced apoptosis of K562 cells in a dose-and time-dependent manner. Coincidently, the protein phosphorylation on tyrosine residues was reduced and the expressions of anti-apoptotic protein Mcl-1 and Bcl-xl were down-regulated after exposure to STI571. It is concluded that STI571 induces the apoptosis of CML cells by down-regulating the expressions of Mcl-1 and Bcl-xl, which suggests that Mcl-1 and Bcl-xl may play an important role in anti-apoptotic process of CML cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Benzamides ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Imatinib Mesylate ; K562 Cells ; Myeloid Cell Leukemia Sequence 1 Protein ; Phosphorylation ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrimidines ; pharmacology ; bcl-X Protein ; metabolism

12E7 Antigen ; Adult ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; pathology ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Nephrectomy ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; Wilms Tumor ; pathology

Migration of donor T cells to the host second lymphoid organs and homing of activated donor T cells into the target tissues play crucial role in the pathophysiology of acute graft-versus-host disease (aGVHD). More deep understanding of the mechanisms for donor T cell migration and homing reveals an important significance in preventing the initiation of aGVHD. In this review, the migration of donor T cells to host second lymphoid organs, homing of donor activated T cells into the target organs, homing of regulating T cells into target organs, the mechanisms of T cell migration and homing in process of aGVHD and its study prospects were summarised.
Cell Movement ; Graft vs Host Disease ; Humans ; Receptors, Lymphocyte Homing ; T-Lymphocytes ; cytology ; immunology

8 ?g/ml were 7 strains in prophylactic treatment group and 3 strains in non-fluconazole prophylactic treatment group respectively.The two groups had significant difference (x~2=8.75,P

Objective To investigation the pathological characteristics of esophageal squamous cell carcinoma to provide reference criteria for delineating the target area in radiotherapy.Methods Fifty-two patients from the Fourth Hospital of HeBei Medical University underwent resection whom all had been proved to have esophageal squamous cell carcinoma before operation.Chest CT was scanned and transmitted to the 3- dimensional conformal planning system for radiotherapy by VRX-16 scanner.The lesion of esophageal carcinoma was delineated in the 3-dimensional rebuild CT image and the lesion volume was computed by digital rebuild program.Every surgically resected specimen was made into pathologic giant section.The actual size of the specimen was obtained by calculating the size under the microscope with the shrinkage ratio.Multicentric carcinomatous lesion,severe dysplasia and direct intramural infiltration were observed in the giant section with a microscope and the order of such pathological characteristics were analysed statistically.Results 1.The tumor length by different method of preparation of operated specimens differed obviously.The longest was shown by CT. 2.Multicentric carcinomatous lesion was found in 15(29%)cases out of 52 patients.Proximal to the tumor,the mean distance between the multicentric carcinomatous lesion and the main lesion plus the length of the multicentric carcinomatous lesion was 3.02?1.45cm.Distal to the tumor,it was 2.60?2.44 cm.Severe dysplasia was found in 28 patients.Proximally,the mean distance between the severe dysplasia and the main lesion plus the length of the severe dysplasia was 2.45?1.30 cm.Distal to the tumor,it was 3.24?2.19 cm.Direct intramural infiltration was found in 41 patients,of which the mean length being 2.80?1.52 cm proximally and 2.02?1.51 cm distally. 3.Tumor thrombus was found in 6 patients and lymphoduct infiltration in 36 patients.Direct intramural infiltration was found at higher incidence in specimens complicated with lymphoduct infiltration(86%)and those complicated with tumor thrombus(91%).There were no apparent factors affecting severe dysplasia.The proximal distance to direct intra- mural infiltration was much longer than distally.Conclusions Multicentric carcinomatous lesion,severe dysplasia and direct intramural infiltration may be observed in esophageal squamous cell carcinoma.Multicentric carcinomatous lesion and direct intramural infiltration are obviously correlated with lymphoduct infiltration.To cover 95% of the microscopic extension,a margin of 5.0 cm is needed proximal to the base of gross tumor volume,and 7.5 cm distal to it.To cover 90% of the microscopic extension,a margin of 4.5 cm is needed proximally,and 5.0 cm distally.