Objective To compare the incidence of C5 nerve root palsy after laminoplasty and laminectomy with internal fixation for treating multilevel cervical spondylotic myelopathy (MCSM).Methods From January 2008 to August 2012,98 patients with MCSM were treated with laminoplasty (47 patients,group A) or laminectomy (51 patients,group B) with internal fixation.All the patients were followed up for 13-56(26.5 ± 7.9) months.In both groups,Cobb's method was applied to measure cervical lordotic angle,and Ishihara's method was conducted to measure cervical curvature index (CCI) before and after operation.The incidence of C5 nerveroot palsy was recorded and compared.Results The incidence of C5 nerve root palsy in group A was 2.1％ (1/47),while 21.6 ％ (11/51) in group B (x2 =5.430,P ＜ 0.05).The JOA scores in group A and group B before and after operation and improvement rate of JOA scores had no significant difference (P＞ 0.05).The cervical lordotic angle and CCI in group A and group B before and after operation had no significant difference (P ＞ 0.05).The improvement rate of CCI between two groups had no significant difference (P ＞ 0.05).All of 11 patients with C5 nerve root palsy were group B 1,and other 40 patients were group B2.The improvement rate of CCI in group B1 was significantly higher than that in group B2 [(38.7 ± 18.3)％ vs.(22.1 ± 12.1)％](t =1.772,P＜ 0.05).Conclusions Compared with laminoplasty,laminectomy with internal fixation has a higher incidence of C5 nerve root palsy.The C5 nerve root palsy may be associated with postoperative increase of cervical lordosis angle.Moreover,tethering of the C5 root may he one of its important pathomechanisms.

As unrelated allogeneic umbilical cord blood transplantation (UCBT) has been developed for 20 years already since 1988, more than ten thousands cases have cumulatively undergone UCBT over the world. A huge number of clinical data confirmed that UCBT had unique characters with low rate of severe GVHD. The efficacy and data on TRM, relapse and EFS of allogeneic UCBT with HLA 0-1 mismatched are similar to those in HLA matched BMT. UCBT has become the optimal choice for source of hematopoietic stem cells for allogeneic stem cell transplant especially when HLA-matched or haploidentical donors are not available in time. In most developed countries, unrelated allogeneic UCBT developed successively, and in recent years HLA mismatched UCBT with double units performed in adults increased even more rapidly than in children. Another recent trend of UCBT has been extending to treat some non-malignant but refractory diseases in pediatrics, such as severe combined immunodeficiency, thalassemia major, bone marrow failure syndrome and metabolic disorders. The clinical successful practice of double units for cord blood transplantation inspires to ponder over questions remaining mystery. What is the conflict like between two mismatched donor cells in vivo, which does not spoil the whole transplantation but enable the patient to be engrafted successfully without any increment of the dosage by the sum of two doses together? How can they both be taken at the same time firstly by the recipient, but why does only one predominate later? What are the factors enable the donor cells of the winner to sustain? With the references of the international experiences, how to solve the clinical encountered problems, perspective of unrelated allogeneic UCBT and proper strategies to be enacted are reviewed.
Cord Blood Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Tissue Donors ; Transplantation, Homologous

Manifold methods for bone mineral density analyses are introduced in this paper, and the characteristics of precision, accuracy, position, convenience, sensitivity and the radiation hazards of these methods are also discussed here.
Absorptiometry, Photon ; methods ; Bone Density ; Humans ; Osteoporosis ; diagnosis ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; Ultrasonography ; instrumentation

OBJECTIVETo investigate the molecular mechanisms underlying in the treatment of inflammatory bowel disease by Pulsatilla Decoction.

METHODSForty Wistar male rats were randomly divided into 5 groups( n = 8)control group, model group, model + positive control group (mesalazine), Pulsatilla Decoction treatment group, in addition, the Pulsatilla Decoction treatment group was divided into middle and high dose group. Intragastric administration was used in the positive control group and Pulsatilla Decoction treatment group. The expression of interleukin-1beta (IL-1beta), interleukin-6(IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by real time PCR after extraction of RNA from colons.

RESULTSCompared with the model group, positive medicine and Pulsatilla Decoction group, especially high-dose group, could effectively inhibit the expression of IL-1beta, IL-6 and TNF-alpha.

CONCLUSIONPulsatilla Decoction could exert its effect in the treatment of inflammatory bowel disease by inhibiting the expression of proinflammatory cytokines.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Inflammatory Bowel Diseases ; drug therapy ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Interleukin-6 ; genetics ; metabolism ; Male ; Phytotherapy ; Pulsatilla ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Adolescent ; Adult ; Female ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Tibia ; injuries ; surgery ; Tibial Fractures ; surgery ; Young Adult

The aim of this study was to investigate the biological characteristics of human bone marrow mesenchymal stem cells from bone marrow of different age donors. The experiments were divided into four groups by donors age, group A represented MSC derived from fetal bone marrow, group B represented MSC derived from bone marrow of 0-20 years old donors, group C represented MSC derived from bone marrow of 20-40 years old donors and group D represented MSC derived from bone marrow of donors older than 40. The growth, purification, proliferation and multipotential abilities of MSC in 4 groups were observed and their immunophenotypes were determined by flow-cytometry. The level of cytokines (IL-6, SCF, FLT-3L, SDF-1 and TGF-beta1) were assayed by ELISA method. Cell cycles were analyzed to show the proliferation index (PrI). MSCs derived from bone marrow of 4 groups were injected subcutaneous into NOD/SCID mouse to observe the safety. The results showed that different age donors bone marrow all gave rise to MSC. These cells were similar in morphology, antigenic phenotype, differentiation potential and cell cycle. The primary culture time of group B was shorter than other groups. The duration of passage 1 (P1) was 5.5 days, and the duration of P10 was 33 days, after P10 culture, (5.19 +/- 2.15) x 10(10) MSCs were obtained from 8 x 10(6) MNC of this group. The primary culture time of groups A, C, D were longer, the duration of P1 were 15, 7 and 13 days for group A, C and D respectively, and the duration of P10 was 50, 60 and 72 days for group A, C and D, respectively. After P10 culture, (4.98 +/- 2.08) x 10(10), (1.86 +/- 0.47) x 10(10), (0.64 +/- 0.22) x 10(10) MSCs were obtained from 8 x 10(6) MNC of group A, C and D respectively. The morphology of MSC of group A was longer and slender. The ability of expansion decreased after P15 for A group, P10 for B group and P8 for C and D groups. The levels of SCF, FLT3-L, IL-6 and SDF-1 in group B were higher than other groups. Karyotype analysis showed that MSCs from 4 groups were normal, and tumor-like tissues were not developed after cultured MSCs were inoculated in NOD/SCID mice. It is concluded that there was relationship between age and the biological characteristics of human bone marrow mesenchymal stem cells. For clinical use, especially in hematopoietic stem cell transplantation (HSCT), 0-20 years old donors were perfect MSCs donors who can provide sufficient MSCs in relatively short times. MSCs of group B can be used as stem cell source because the biological characteristics of MSCs of groups B are superior to that of other groups.
Adolescent ; Adult ; Age Factors ; Animals ; Blood Donors ; Bone Marrow Cells ; cytology ; immunology ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Child ; Child, Preschool ; Cytokines ; analysis ; Enzyme-Linked Immunosorbent Assay ; Fetus ; Flow Cytometry ; Humans ; Immunophenotyping ; Infant ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; immunology ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Transplantation, Heterologous

Transmembrane protein p185 (the product of Her2/c-erbB-2 gene) is a member of the epidermal growth factor receptor (EGFR) family. Its overexpression was found in about 30% of breast cancer. It is essential to obtain soluble extracellular domain (ECD) of p185, especially disulfide bond rich domains, for identifying the epitopes of anti-p185 antibodies and researching the interrelationship between the antigen and antibody. The disulfide bond rich domain I-II and domain IV of p185 ECD were amplified from plasmid pBabe/erbB-2 by PCR respectively. These two fragments were inserted into pGEX/4T-1 vector, transfected into E. coli Origami B (DE3) pLysS and expressed inductively by low concentration of IPTG and low temperature overnight. After the pressure lysis of cells, the supernatants were analyzed by SDS-PAGE and the result demonstrated that this GST-fusion protein was expressed solubly in the amount of 10-15 mg/L. By the ELISA, Western blot and other immunological assays, the fusion proteins and their GST cut-off derivates both showed binding activities with several anti-p185 antibodies respectively. These results indicated that it was a feasible and effectual method to express disulfide bond rich domain I-II and domain IV of p185 ECD and this method may also be used to express other disulfide bond rich proteins.
Antibodies, Monoclonal ; immunology ; Disulfides ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Receptor, ErbB-2 ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Transfection

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

OBJECTIVETo observe the in vivo distribution of mesenchymal stem cells (MSCs) after administrated by intra-bone marrow (IBM) or intravenous (i.v.), and compare the effects on hematopoiesis reconstitution and GVHD in rat BMT models.

METHODS(1) MSCs from male Wistar rats marked with CFSE were injected into the bone marrow cavity (IBM) or the vein (i.v.) of recipient rats, and observed the distribution of MSCs in vivo. (2) Allogeneic BMT model of Fischer344 rats (RT1A(1)) to Wistar rats (RT1A(u)) was established. The recipient rats were exposed to 8 Gy of gamma irradiation 1 day before transplantation. The 6 groups were (1) IBM group [IBM-injection of MSCs + IV-injection of bone marrow cells (BMC)]; (2) IV group (i.v.-injection of MSCs (i.v.) + i.v.-injection of BMC); (3) BMT group (only i.v.-injection of BMC); (4) MSCs control group (only i.v.-injection of MSC); (5) normal control group and (6) irradiation control group.

RESULTS(1) After i.v.-injection, large numbers of the MSCs lodged in lungs while small numbers in the peripheral blood, liver, thymus and spleen, and a few marked MSCs could be seen in bone marrow. After IBM injection, most cells distributed in long bones and those lungs were less than that in i.v. group. (2) Co-transplantation of MSCs (IBM/IV) could accelerate the recovery of hematopoiesis, including the recovery of WBC, hemoglobin and platelet, and in IBM-injection was more effective in the recovery of hematopoiesis than that in i.v. group. (3) Incidence rate of GVHD in BMT group was 42% (3/7), and no GVHD occurred in co-transplantation groups. (4) Recovery of CFU-Mix and CFU-MSCs could be seen at 21st and 30th day after transplantation in co-transplantation groups, and IBM-injection was more effective than i.v.-injection.

CONCLUSION(1) IBM-injection results in most MSCs distributed in long bones. (2) MSCs improve the survival rate after BMT. (3) Co-transplantation of MSCs accelerates the recovery of hematopoiesis and reduces the morbidity of GVHD. (4) MSC promotes reconstitution of hematopoietic cells and bone marrow MSCs in recipient rates and the effects of MSCs administrated via IBM is more effective than via i.v.

Animals ; Bone Marrow Transplantation ; Female ; Graft vs Host Disease ; prevention & control ; Hematopoiesis ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Models, Animal ; Rats ; Rats, Wistar ; Transplantation, Homologous

OBJECTIVETo explore the most efficient culture system which can induce cord blood (CB)-mononuclear cells (MNC) to differentiate into mature T/NK cells in vitro.

METHODSThe CB MNCs were cultured in six culture systems respectively for 4 weeks. The T/NK cell surface phenotypes were analyzed by flow cytometry and the absolute numbers of nucleated cells (NCs) were counted at each time point. Moreover, cell morphology was identified by Giemsa-Wright staining, and cytotoxicity of the cultured cells to K562 and Raji tumor cells was also evaluated by MTT method.

RESULTSCultured in the cytokine cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, the NCs were (20 approximately 26) x 10(6)/ml in numbers at day 22. The percentage of lymphocytes in the NCs and that of CD(3)(+) T cells in the lymphocytes both exceeded 90% at the same time. Most of the CD(3)(+) T cells were CD(3)(+)CD(8)(+) and the percentage of CD(3)(+)CD(4)(+) T cells declined gradually. The percentage of CD(3)(+)CD(56)(+) NKT cells and gamma delta(+)T cells in the lymphocytes arised from lower than 2% to 30% approximately 40% and 10% approximately 15%, respectively. CD(3)(-)CD(56)(+) NK cells were not expanded. The cytotoxic activity of the cultured cells to K562 and Raji cells at an effector:target (E:T) ratio of 50:1 was over 75% and about 32% approximately 65%, respectively.

CONCLUSIONThe most efficient culture system which can induce CB MNC to differentiate into mature T/NK cells in vitro is the cytokines cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, and the optimum culture time is 22 days.

Cell Differentiation ; Cell Division ; Cytotoxicity, Immunologic ; Fetal Blood ; cytology ; Humans ; Infant, Newborn ; Interleukin-2 ; pharmacology ; Killer Cells, Natural ; cytology ; Leukocytes, Mononuclear ; cytology ; Receptors, Antigen, T-Cell, gamma-delta ; analysis ; T-Lymphocytes ; cytology ; Tumor Necrosis Factor-alpha ; pharmacology