To explore the efficacy of the different combinations of stem cell factor (SCF), FLT3 ligand (FL) and interleukin (IL) 2, 7, 15 on the induction and expansion of cord blood (CB) derived CIK/ NK cells in vitro, according to the different combinations of cytokines, combinations of cytokines were divided into 3 groups: group A (SCF + IL2 + IL7 + IL15), group B (SCF + FL + IL2 + IL7 + IL15) and group C (IL2 + IL7 + IL15 as control group). At 21 days of culture, the proportion and the expansion rate of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) NK cells were measured. The results showed that at 21 days in culture, the percentages of CD3(+)CD56(+) CIK cells derived from CB-MNC in group A, B and C were (19.84 +/- 2.93)%, (26.20 +/- 4.05)%, (24.03 +/- 4.99%) and that of NK cells were (49.60 +/- 1.40)%, (51.16 +/- 6.45)% and (30.85 +/- 8.12)%, respectively. The proliferation of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) NK cells was the most effective in group B (SCF + FL + IL2 + IL7 + IL15). Expansion multiple of CIK cell number increased from 575.81 +/- 221.72 (control) to 796.09 +/- 278.47 with peak value of 1 313, and that of NK cells increased from 30.39 +/- 14.47 (control) to 65.85 +/- 30.83 with peak value of 121.06. In conclusion, a proper cytokines (SCF + FL + IL2 + IL7 + IL15) culture system can effectively induce and expand CB CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) NK cells simultaneously.
CD3 Complex ; analysis ; CD56 Antigen ; analysis ; Cytokines ; pharmacology ; Fetal Blood ; cytology ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukin-7 ; pharmacology ; Killer Cells, Natural ; drug effects ; Membrane Proteins ; pharmacology ; Stem Cell Factor ; pharmacology

To screen and separate the genes differentially expressed in human embryonic aorta-gonad-mesonephros (AGM)-derived stromal cells, a subtracted library was generated through the suppression subtractive hybridization using the cDNA of human embryonic AGM-derived stromal cells as target and human fetal liver (FL)-derived stromal cells as drivers. Then a high though screening technique, gene chip, was used to screen the differentially expressed genes in the established subtractive library. Approximately 18 of the resulting subtracted cDNA clones were partially sequenced and analyzed by blastn in the GenBank database. The results showed that 211 Clones were selected and identified from the established subtractive library, the positive ratio was amount to 76.4%. 18 over-expressed genes were screened by gene chip with more than a 5-fold difference expression levels between AGM and FL-derived stromal cells, and were selected to sequence, results of sequencing indicated that the 18 sequences was compared to known sequences in the GenBank database, and among the sequenced clones, 14 sequences were considered as part of the known genes, and 4 sequences representing previously unknown genes. The known genes were reported to involve the regulation of cell migration, cell differentiation, cell proliferation, cell cycle, signal transduction, and angiogenesis. Most of these genes have not been reported to relate to the haematogenesis in ontogeny. It is concluded that many genes both known and unknown are differentially expressed in human embryonic aorta-gonad-mesonephros-derived stromal cells. Discovery of these genes provides a solid foundation to elucidate the mechanism of haematogenesis in ontogeny.
Aorta ; embryology ; Cloning, Molecular ; Embryonic Stem Cells ; cytology ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Gene Expression ; Gene Expression Profiling ; Gonads ; embryology ; Hematopoiesis ; physiology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesonephros ; cytology ; Oligonucleotide Array Sequence Analysis ; Stromal Cells ; cytology ; metabolism

OBJECTIVETo explore whether the co-transplantation of mesenchymal stem cells (MSC) from human umbilical cord blood (UCB) with UCB-derived CD34(+) cells in NOD/SCID mice could promote engraftment and accelerate hematopoiesis recovery.

METHODSAfter sublethal irradiated ((60)Co 2.5 Gy), NOD/SCID mice received within 24 hours UCB CD34(+) cells (1 x 10(5) per mouse for low dosage group or 1 x 10(6) per mouse for high dosage group) with or without human UCB-derived MSC (1 x 10(6) per mouse) transplantation by lateral tail vein injection. Peripheral blood cells of transplanted mice were measured for white blood cell count, hemoglobin and platelet count at 10th, 20th, 30th, 40th and 56th day. At the end of 8th week after transplantation, all the alive mice were sacrificed and human derived CD45(+), CD45(+)CD3(+), CD45(+)CD19(+), CD45(+)CD33(+) cells in the bone marrow (BM) were assayed by flow cytometry.

RESULTS(1) In the low dosage group, co-transplantation of MSC significantly raised the engraftment rate (26.02% vs 16.52%) (P < 0.05). (2) The survival rate in high dosage group was 80% for co-transplantation mice and 70% for CD34(+) cells alone transplantation mice. The survival rate in low dosage group was 70% for co-transplantation mice and 50% in CD34(+) cells transplantation mice. (3) In both dosages groups co-transplantation accelerated the hematopoiesis recovery. (4) At the end of 8 weeks after transplantation, in low dosage group, CD45(+)CD33(+) and CD45(+)CD19(+) cells were more in co-transplantation mice than in CD34(+) cells alone transplantation mice, but in high dosage group, the percentage of these two kinds of cells had no difference. In both dosage groups the percentage of CD45(+)CD41a(+) cells were higher in co-transplantation than in transplantation alone mice. CD45(+)CD3(+) cells were low in all groups.

CONCLUSIONS(1) In low dosage transplantation, human UCB MSC could promote human CD34(+) cells engraftment in transplanted mice. (2) Co-transplantation of human UCB MSC and human UCB CD34(+) cells could significantly promote the hematopoiesis reconstitution and improve the survival rate of NOD/SCID mice. (3) MSC could promote human UCB CD34(+) cells to differentiated into B-lymphocytes, granulocyte and megakaryocyte in vivo.

Animals ; Antigens, CD34 ; Cord Blood Stem Cell Transplantation ; Female ; Fetal Blood ; cytology ; Hematopoiesis ; Humans ; Mesenchymal Stem Cell Transplantation ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Transplantation, Heterologous

OBJECTIVESTo evaluate the effects of K562-dendritic cell (DC) and Raji-DC fusion vaccines on the cytotoxicity of cord blood (CB) derived cytokine-induced killer/natural killer (CIK/NK) cells.

METHODSDC and CIK/NK cells were derived from CB mononuclear cells. CB-DC were fused with inactivated K562 or Raji cells by PEG to form K562 or Raji-DC fusion vaccine. The CIK/NK cells stimulated by different co-culture antigens were three groups: K562-DC or Raji-DC fusion vaccine group, inactivated K562 or Raji plus DC group, and CB-DC alone group. The cytotoxicity of CIK/NK cells stimulated by different co-culture antigens was measured by MTT test.

RESULTSAll the antigens used for stimulation could enhance the cytotoxicity of CB-CIK/NK cells, with no specificity difference. At 20:1 effector-target ratio, the cytolytic activities of K562-DC and Raji-DC fusion vaccine groups against Raji cells were (75.44 +/- 4.19)% and (81.33 +/- 4.18)% respectively (P < 0.05); and that of inactivated K562 + DC and Raji + DC group against Raji cells were (73.12 +/- 4.22)% and (80.49 +/- 4.27)%, respectively (P < 0.01). There was no significant difference in the cytotoxicity to K562 cells between the two fusion vaccine groups (P > 0.05). The cytotoxicity of CB-CIK/NK cells immunized by Raji cells was higher than that by K562 cells. In CIK/NK cells co-stimulated by the same tumor antigen, there was no significant difference in the cytotoxicity between DC fusion vaccine group and inactivated cells plus DC group to different tumor cells.

CONCLUSIONSThe cytotoxicity of CB-CIK/NK cells to tumor cells was not specific. There was no significant difference in the cytotoxic activity of CB-CIK/NK cells between the DC fusion vaccine group and inactivated cells plus DC group.

Cancer Vaccines ; immunology ; pharmacology ; Cells, Cultured ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; cytology ; Humans ; K562 Cells ; Killer Cells, Lymphokine-Activated ; immunology ; Killer Cells, Natural ; immunology

BACKGROUNDPatients with myelodysplastic syndrome (MDS) display a very diverse pattern. In this study, we investigated prognostic factors and survival rate in adult patients with MDS refractory anaemia (MDS-RA) diagnosed according to French-American-British classification and evaluated the International Prognostic Scoring System (IPSS) for Chinese patients.

METHODSA multi-center study on diagnosis of MDS-RA was conducted to characterize the clinical features of Chinese MDS patients. The morphological criteria for the diagnosis of MDS-RA were first standardized. Clinical data of 307 MDS-RA patients collected from Shanghai, Suzhou and Beijing from 1995 to 2006 were analyzed using Kaplan-Meier curve, log rank and Cox regression model.

RESULTSThe median age of 307 MDS-RA cases was 52 years. The frequency of 2 or 3 lineage cytopenias was 85.6%. Abnormal karyotype occurred in 35.7% of 235 patients. There were 165 cases (70.2%) in the good IPSS cytogenetic subgroup, 44 cases (18.7%) intermediate and 26 cases (11.1%) poor. IPSS showed 20 (8.5%) categorized as low risk, 195 cases (83.0%) as intermediate-I risk and 20 cases (8.5%) as intermediate-II risk. The 1-, 2-, 3-, 4- and 5-year survival rates were 90.8%, 85.7%, 82.9%, 74.9% and 71.2% respectively. Fifteen cases (4.9%) transformed to acute myeloid leukaemia (median time 15.9 months, range 3 - 102 months). Lower white blood cell count (< 1.5 x 10(9)/L), platelet count (< 30 x 10(9)/L) and cytogenetic abnormalities were independent prognostic factors by multivariate analysis, but age (= 65 years), IPSS cytogenetic subgroup and IPSS risk subgroup were not independent prognostic factors associated with survival time.

CONCLUSIONSChinese patients were younger, and had lower incidence of cytogenetic abnormalities, more severe cytopenias but a more favourable prognosis than Western patients. The major prognostic factors were lower white blood cell count, lower platelet count and fewer abnormal karyotypes. The international prognostic scoring system risk group was not an independent prognostic factor for Chinese myelodysplastic syndrome patients with refractory anaemia patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Refractory ; etiology ; mortality ; Asian Continental Ancestry Group ; Child ; China ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; mortality ; Prognosis

Objective To investigate the prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong province.Methods Data from 169 218pregnant women in different regions of Guangdong province from January 2005 to June 2010 were analyzed retrospectively.The prevalence and epidemiological characteristics of thromboembolic disease during pregnancy or puerperium were investigated.Results Of the studied population,( 1 )20 l cases ( 1.3‰ ) suffered from thromboembolic disease during pregnancy or puerperium including 128 cases of deep vein thrombosis (DVT),68 cases of cerebral venous thrombosis (CVT) and 5pulmonary embolism,the prevalence rates were 0.8‰,0.4‰,and 0.02‰ respectively.(2) Risk factors in different regions showed that,in the Pearl River Delta area,the major risk factors for DVT would include previous or family history of thrombosis,pregnancy complications,with medically involved diseases,prolonged bed rest and pregnancy weight gain ＞ 15 kg etc.While in castern,western,northern parts of Guangdong,the major risk factors for DVT would include pregnancy weight gain ＞ 15 kg,prolonged bed rest,preeclampsia,cesarean section and complications during pregnancy.In Pearl River Delta region,the major risk factors for CVT would include eclampsia,preeclampsia,pregnancy complications,prolonged bed rest ＞3 days,past history or family history of thrombosis.While eclampsia,preeclampsia,advanced age or younger age,pregnancy weight gain ＞15 kg,complications during pregnancy were the major risk factors for CVT in the eastern,western or northem parts of Guangdong.Conclusion Prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong were different.It was crucial to take effective measures in pregnant women with different epidemiological characteristics and risk factors to prevent and reduce the incidence of peripartum thromboembolic disease.

OBJECTIVETo summarize and critically assess the effificacy of Eastern and Western manipulative therapies for the treatment of neck pain in adults.

METHODSA search of PubMed/MEDLINE, the Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, EMBASE, etc. from their inception date to January 2014 with Chinese, Japanese, and Korean databases. Two reviewers independently selected randomized controlled trials (RCTs) with negative control or blank control, extracted data and assessed methodological quality. Meta-analysis and levels of evidence were performed by Revman5.1 and Grades of Recommendations Assessment, Development and Evaluation (GRADE) approach.

RESULTSNineteen clinical trials with adequate randomization were included in this review, 11 of them had a low risk of bias. The primary outcome for short-term pain had no significant differences, however, the secondary outcome, only the Numerical Pain Rating Scale (NPRS) score of intermediate-term [n=916, pooled mean differences (MD) =-0.29, P=0.02], the Neck Disability Index (NDI) score of short-term (n=1,145, pooled MD=-2.10, P<0.01), and intermediate-term (n=987, pooled MD=-1.45, P=0.01) were signifificantly reduced with moderate quality evidence. However, it supported the minimally clinically important difference (MCID) of the Visual Analogue Scale and NPRS pain score to be 13 mm, while NDI was 3.5 points. The meta-analysis only suggested a trend in favor of manipulative therapy rather than clinical signifificance.

CONCLUSIONSThe results do not support the existing evidences for the clinical value of Eastern or Western manipulative therapy for neck pain of short-term follow-up according to MCIDs. The limitations of our review related to blinding, allocation concealment and small sample size.