Objective To assess the association between NOD2 gene single nucleotide polymorphisms (SNPs) and leprosy in Chinese Yi population.Methods Whole blood samples were obtained from 300 patients with leprosy and 300 healthy human controls of Yi nationality in Sichuan province.Genomic DNA was extracted,and a SNaPshot assay was performed to determine the genotypes of four single nucleotide polymorphisms (SNPs) of the NOD2 gene,including rs9302752,rs7194886,rs8057341 and rs3135499.Chi-square test was conducted to compare allele frequency,and Hardy-Weinberg equilibrium was tested.Results The genotype distribution of all the four SNPs was consistent with Hardy-Weinberg equilibrium (all P ＞ 0.05).Significant differences were observed between the patients with leprosy and healthy controls in both genotype distribution and allele frequency of the SNP rs3135499 (both P ＜ 0.01),but not in those of the other three SNPs (all P ＞ 0.05).Conclusion The SNP rs3135499 of the NOD2 gene may be associated with the development of leprosy in Chinese Yi population.

The object of this study was to explore the effect of FL gene transfected marrow stromal cells on expansion of human cord blood CD34(+) cells ex vivo. A co-culture system was established with FL gene transfected human marrow stromal cells and cord blood CD34(+) cells. The CD34(+) cells were cultured in different culture systems with different combinations of stromal cells and SCF, IL-3 and FL. The numbers of total nucleated cells, CFCs and CD34(+) cells were repeatedly counted in culture systems for 3 weeks. The results showed that the FL transgenic marrow stromal cell feeder layer significantly enhanced the expansion of total nucleated cells, CFC and CD34(+) cells, compared with stromal cell feeder layer and stromal cell-free culture systems. The transgenic stromal cells and cytokines co-culture system manifested the most potent expansion of the cord blood cells with increase of the number of total nucleated cells by (122.5 +/- 4.3)-fold, CFCs by (39.6 +/- 2.7)-fold and CD34(+) cells by (11.8 +/- 0.52)-fold. It is concluded that human umbilical cord blood CD34(+) cells can be extensively expanded ex vivo by using FL gene transfected stromal cells combined with cytokines.
Antigens, CD34 ; analysis ; Cell Separation ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Interleukin-3 ; pharmacology ; Membrane Proteins ; physiology ; Stem Cell Factor ; pharmacology ; Transfection

To study the effect of mesenchymal stem cell (MSC) on immune function, MSCs were isolated and cultured from human bone marrow cells. The purity of MSCs were identified with the spindle-fibroblastic morphology characterization by microphotograph and the phenotypes were tested by flow cytometry. MSCs were plated in 96-well plates (2,000/well and 1,000/well), and cocultured for 3 days with T cells isolated from cord blood. Cord blood T cells non-cocultured with MSC acted as control group. After cord blood T cells stimulated by PHA for 60 hours, [(3)H]-thymidine was added to each well and T cell proliferation was assessed by [(3)H] thymidine incorporation. The results showed that cord blood T cell proliferation was suppressed when 2,000 MSCs were plated each well and cord blood T cell proliferation was activated when 1,000 MSCs were plated. Our results suggested that the immunomodulatory function of MSC seemed dependent on cell dose. High concentration of MSC most often resulted in inhibition, while low concentration resulted in stimulation.
Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; Cell Division ; immunology ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Lymphocyte Activation ; drug effects ; immunology ; Mesoderm ; cytology ; Phytohemagglutinins ; pharmacology ; Stem Cells ; cytology ; T-Lymphocytes ; cytology ; drug effects ; metabolism ; Thymidine ; metabolism ; Tritium

This study was purposed to investigate the immunoregulatory effect of endothelial cells derived from mesenchymal stem cells (MSC). The human MSC was induced to differentiate into endothelial cells for one week. The phenotypes were evaluated by flow cytometry, the cell morphologic feature was observed by invert phase-contrast microscope and analysis of capillary formation was performed by using the in vitro angiogenesis kit. The immunoregulatory effect was detected by lymphocyte transformation test. The result indicated that during the differentiation cells grew fast and there was no significant change in the phenotypes, i.e. CD73, CD105, HLA-ABC were positive and CD34, CD80, CD86, HLA-DR, CD31 were negative. Immunofluorescence analysis showed typical expression of the von Willebrand factor. Differentiated MSCs formed capillary-like structure. Endothelial cells derived from MSC also revealed immunosuppressive effect on T cell proliferation in a dose-dependent manner. It is concluded that endothelial cells derived from MSC also harbor immunoregulatory effect on T lymphocytes.
5'-Nucleotidase ; metabolism ; Cell Differentiation ; physiology ; Cells, Cultured ; Child ; Endothelial Cells ; cytology ; immunology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; T-Lymphocytes ; immunology ; von Willebrand Factor ; metabolism

Long-tem culture initiating cells(LTC-IC), and in vitro assay of hematopoietic stem/progenitor cells, still represent a heterogeneous population in terms of proliferative capacity and sensitivity to different growth factors. Human umbilical cord (CB) is rich of hematopoietic progenitor cells measured by clonogenic assays and stem cells capable of reconstituting the marrow after transplantation. The influence of culture conditions on the in vitro behavior of LTC-IC from CB was evaluated. First, by using IRES sequence, FL and TPO cDNA were recombined with retroviral vector pLXSN by gene recombination technology. The recombinant plasmid pLFSN, pLTSN, pLFTSN were transfected into human stromal cell line HFCL. Then, LTC-IC were evaluated in long term cultures, comparing five types of stromal feeder layers: human bone marrow stromal cell, human stromal cell line HFCL, and stromal cell lines HDF tranfected with FL gene, HLT transfected with TPO gene or HFT co-transfected with FL and TPO genes. The results were demonstrated that after 8 weeks of coculture, three types of stromal cell lines that supported the maintenance of CFU-C for up to 3 weeks in vitro were identified. However, cocultivation of human bone marrow stromal cell and CB CD34(+) cells on HFT, CFU-C production continued up to 6 weeks or longer on these stroma. The absolute LTC-IC frequency in CD34(+) cells on human bone marrow stromal cell (2.65 +/- 0.76/1 000 cells) was no significant difference with on HFT (3.65 +/- 0.58/1 000 cells). Thus, HFT acts by direct contact to maintain the phenotype and function of the most primitive and quiescent human progenitors. Furthermore, HFT cell line was selected as the optimal one for supporting long-term culture feeder. It was concluded that LTC-IC progenitors from cord blood maintain growing upon the FL/TPO gene-modified stromal feeder layers in vitro.

OBJECTIVETo explore the role of diffusion tensor imaging (DTI) in neuronavigation surgery of brain tumors involving pyramidal tracts.

METHODSForty-nine patients with brain tumors involving pyramidal tracts were randomly divided into trial group (DTI navigation) and control group (traditional navigation). The patients in trial group underwent DTI and T1 weighted 3D navigational magnetic resonance imaging (MRI) studies. The main white matter tracts were constructed by the DTI datasets, and merged to the anatomical structure, which was delineated by the T1-weighted three-dimensional fast spoiled gradient recalled sequence (3D/FSPGR). The relationship between the tumors and adjacent pyramidal tracts were segmented and reconstructed for three-dimensional visualization.

RESULTSIn 25 patients of trial group and 24 patients of control group, the statistic analysis confirmed well balance of main variations. The tumors were completely resected in 12 patients (50.0%) of control group and in 20 patients (80.0%) of trial group (P < 0.05). Postoperative aggravated contralateral extremities weakness or hemiplegia due to pyramidal tract injury occurring in 75.0% cases of control group whereas only 20.0% patients in trial group (P < 0.01). The mean Karnofsky scale were 69.58 +/- 23.49 and 84.80 +/- 23.49 respectively in control and trial groups (P < 0.05). The excellent outcome ratio (Karnofsky scale = 90 - 100) was 37.5% in control group and 72.0% in trial group respectively (P < 0.05).

CONCLUSIONSDTI allows individual estimation of large fiber tracts of brain. Furthermore, to integrate spatial three-dimensional information concerning the white matter tracts into traditional neuronavigation images during surgery, was valuable in presenting topographical character of involving (shift or erosive) pyramidal tracts and relationship with the margins of neighboring tumors. The mapping of large fiber tracts was a safe, efficient, reliable technique. DTI should be routinely used in neuronavigation surgery of brain tumor involving pyramidal tracts to plan the optimal trajectory and ensure total resection of the lesions during operation, as well as to decrease potential disability after operation and to shorten the length of hospitalization.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; pathology ; surgery ; Child ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Neuronavigation ; methods ; Pyramidal Tracts ; pathology

OBJECTIVETo isolate and culture human placenta derived adherent cells (hPDAC) and assay their hematopoietic growth factor expression.

METHODSBy enzyme digestion, hPDAC were isolated from human placenta tissue and cultured, and their biological characteristics were studied. The hematopoietic growth factor (HGF) mRNA expression of hPDAC was assayed by RT-PCR.

RESULTShPDAC was successfully isolated from human placenta tissue, which was further confirmed as mesenchymal stem cell-like cells. HGF including SCF, FL, G-CSF, GM-CSF, M-CSF and IL-6 were expressed in hPDAC.

CONCLUSIONhPDAC could be used as feeder layer for umbilical cord blood CD(34)(+) cells ex vivo expansion.

Cells, Cultured ; Female ; Gene Expression ; Granulocyte Colony-Stimulating Factor ; genetics ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Hematopoietic Cell Growth Factors ; genetics ; Humans ; Interleukin-6 ; genetics ; Macrophage Colony-Stimulating Factor ; genetics ; Membrane Proteins ; genetics ; Placenta ; cytology ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics

OBJECTIVETo prepare and evaluate an intravenous emulsion of tanshinone II(A).

METHODSoybean phospholipid mixing with poloxamer 188 was used as emulsifier. Oleic acid and glycerol were used as co-emulsifier and isoosmotic adjusting agent, respectively. The coarse emulsion was first prepared and following homogenization was carried out for the coarse emulsion by using a high pressure homogenizer.

RESULTThe average diameter of the prepared tanshinone II (A) emulsion was 211 nm with a zeta potential of -32. 1 mV. There had no changes of diameter, zeta potential, pH value, content and physical appearance for the tanshinone II (A) emulsion stored at 25 degrees C away from light during one year.

CONCLUSIONThe physicochemical properties of the prepared tanshinone II (A) emulsion was stable, which could meet the requirements of intravenous administration.

Centrifugation ; Diterpenes, Abietane ; Drug Stability ; Drug Storage ; Emulsions ; Hydrogen-Ion Concentration ; Injections, Intravenous ; Particle Size ; Phenanthrenes ; administration & dosage ; chemistry ; Quality Control ; Temperature

Small intestinal obstruction is a common complication of primary gastrointestinal cancer or metastatic cancers.Patients with this condition are often poor candidates for surgical bypasses,and placement of self-expanding metal stent (SEMS) can be technically challenging.In this study,we examined the feasibility of combined application of single-balloon enteroscope (SBE) and colonoscope for SEMS placement in patients with malignant small intestinal obstruction.Thirty-four patients were enrolled in this study,among which 22 patients received SEMS placement by using SBE and colonoscope,while the other 12 patients received conservative medical treatment.The patients were followed up for one year.Stent placernent was technically feasible in 95.5％ (21/22).Clinical improvement was achieved in 86.4％ (19/22).For the 19 clinical success cases,the average time of benefits from a gastric outlet obstruction scoring system (GOOSS) increase ≥1 was 111.9±89.5 days.For the 12 patients receiving conservative medical treatment,no significant improvement in GOOSS score was observed.Moreover,a significant increase of Short-Form-36 health survey score was observed in the 19 patients at time of 30 days after stent placement.By Kaplan-Meier analysis,a significant survival improvement was observed in patients with successful SEMS placement,compared with patients receiving conservative medical treatment.Taken together,combined use of SBE and colonoscope makes endoscopic stent placement feasible in patients with malignant small intestinal obstruction,and patients can benefit from it in terms of prolonged survival and improved quality of life.

OBJECTIVE: To study the characteristics and biomechanical mechanism of riding injuries involving bicycles collided by motor vehicles. METHODS: The real traffic accident cases of bicycles collided by motor vehicles, including the information of scenes, bicycles, motor vehicles, rider wounds and traffic directions, were collected. Retrospective method was used to study these riding injuries. In addition, typical cases were selected to simulate traffic accident courses with computer simulation software, and the dynamic data like acceleration, force, moment were cxtracted to compare with those in the real cases. RESULTS: There were no difference of occurring frequency between cases with or without riding injuries, as well as between one-side-collision and front- or back-collision. The riding injuries seemed less in accidents involving large-scale vehicles. The frequency of riding injuries increased with vehicle speed. The wound location was low on collision side and high on opposite. CONCLUSION Analysis of riding injury characteristic in traffic accidents and their biomechanical mechanism would be helpful for estimation of traffic manner.
Accidents, Traffic ; Area Under Curve ; Bicycling/injuries* ; Biomechanical Phenomena ; Computer Simulation ; Humans ; Leg Injuries/pathology* ; Models, Theoretical ; Motor Vehicles ; Perineum/injuries* ; Retrospective Studies ; Wounds and Injuries/pathology*