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

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

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

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

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

OBJECTIVETo study the clinical features and the incidence of early infectious complications following children hematopoietic stem cells transplantation (HSCT).

METHODSThe clinical data of 29 cases with early infectious complications following HSCT was retrospectively analyzed.

RESULTSThe incidence of early infectious complications following HSCT in 31 children (including 22 cord blood transplantation and 9 peripheral blood stem cells transplantation) was 94% (29/31). The first occurrence of the early infectious complications was at a median of 6 (0 - 22) days, the peak time of incidence was at a median of 4 - 7 days post transplantation. The duration of the first early infectious complications was at a median of 9 (3 - 20) days. The occurrence of the second early infectious complications was at a median of 19 (13 - 27) days. For all of the 29 children, when they developed early infectious complications their absolute neutrophil counts (ANC) were all > 0.5 x 10(9)/L. The most common infectious sites were the digestive tract (oral and gastro-intestinal mucositis) and then the respiratory tract. Gram negative blood infections were quite frequent and Pseudomonas aeruginosa was common in the oral-pharynx discharge cultures. Two children had Mycoplasma pneumonia infections and there were 4 incidences with fever but no definite infectious foci. The incidence and duration of early infectious complications following hematopoietic stem cells transplantation were associated with the duration of neutropenia. The source and the MNCs dose of the graft, the difference of conditioning regimen and GVHD prophylaxis method did not have a significant impact on the incidence and duration of early infectious complications. Antibiotic prophylaxis (including Tienam) could delay the occurrence of the early infections significantly.

CONCLUSIONThe incidence and duration of early infectious complications following hematopoietic stem cells transplantation were directly associated with the duration of neutropenia. Tienam regimen could postpone the early infections incidence and had effect of preventing the early infectious complications.

Adolescent ; Antibiotic Prophylaxis ; Child ; Child, Preschool ; China ; epidemiology ; Cilastatin ; therapeutic use ; Drug Combinations ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Imipenem ; therapeutic use ; Incidence ; Infection ; drug therapy ; epidemiology ; etiology ; Leukemia ; therapy ; Male ; Retrospective Studies ; Time Factors

Objective To study the influences of Yang-warming, Blood-activating, and Toxin-removing Recipe(YBTR)-containing serum sampled under different conditions on the proliferation of human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultured in vitro for the experiment. Pharmacological serum for the experiment was prepared as follows: Thirty-two SD rats (male in half) were randomly divided into 4 groups, namely normal saline group and low-, middle-, and high-dose YBTR groups (at the intragastric dosage of 10.35, 31.05, 93.15 g·kg-1 respectively, twice a day). The pharmacological serum was taken from one female rat and one male rat in various groups at 4 time points, i.e. at hour 1, 2 after first intragastric administration on the fourth feeding day, and at hour 1, 2 after first intragastric administration on the sixth feeding day(abbreviated as D3H1, D3H2, D5H1, D5H2 respectively). The effects of YBTR-containing serum on the proliferation of HUVECs were observed by CCK-8 assay method. Results The difference of proliferation-inhibition rate of HUVECs was statistically significant after treated with YBTR-containing serum prepared from rats of different genders at different time(Pgender=0.000<0.01, Ptime=0.000<0.01). The difference of interaction of time and gender was also significant (Ptime × gender=0.001<0.01), and the effect at D3H1 and D5H1 varied with the gender (PD3H2×gender = 0.000 < 0.01, PD5H1×gender = 0.002 < 0.01). The inhibitory action of YBTR-containing serum became stronger with the increase of the dosage of serum of female rat at D3H2, and the inter-group difference was statistically significant (P < 0.05), the effect showing concentration-dependent tendency. The inhibition of HUVECs proliferation reached the peak after treated by various doses of YBTR-containing serum from the female rat at D3H2, while the inhibition arrived to the peak after treated by low- and middle-dose YBTR-containing serum from the male rat at D5H1, and the inhibition arrived to the peak in the group of high-dose YBTR-containing serum from the male rat at D3H1. Conclusion The inhibitory action of YBTR-containing serum on the proliferation of HUVECs was stronger when the serum was taken from the female rat at D3H2.

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.