[Objective]To introduce a method to obtain Schwann cells from newly born SD rats massively and purely by immunomagnetic heads method.[Method]SD rats that had been born within 5 to 7 days were used. Their bilateral sciatic nerves were dissected under sterile condition. Under 16?microscope the nerve fascicles and the epineurium were carefully extracted in oder to get the nerve tract without impurity cell. Then the nerve tract was cut into small particle about 1 mm3.Two enzymes (0.25% trypsinase and 0.2% collagenaseⅠ)were used to digest the particle of sciatic nerve specimens twice. After using 20% fetal bovine serum to stop the process of digestion,centrifugate(1000r/min,5 min) and DF12 culture medium was added into the precipitation.Seven days later,Schwann cells were purified with immunomagnetic heads.During the whole process,the change of Schwann cells was observed under the inverted biological microscop and vital force of Schwann cells was evaluated.The growth curve of Schwann cell was drawn with MTT.The Schwann cells was identicated under immunofluorescent test and record the purity.[Result]The cell separated from the SD rat's sciatic nerve and cultured in incubator was affirmed as Schwann cells.Immunomagnetic heads can be used to purify the Schwann cells.The 96% vigor and 98% pure Schwann cell cultures were generated and passaged after two days.[Conclusion]This method can obtain massive purified normal schwann cells to satisfy the need of tissue-engineered bioartificial nerve graft.

[Objective] To investigate the combined effects of different neurotrophin couples of NGF,bFGF,BDNF on rat spinal cord neurons.[Method]Spinal cord neurons were obtained from SD rats born within 1d,and then were seeded in culture plates.Different factors and their couples were added in each chamber while DMEM/F12 served as controls,concentration of NGF,bFGF or BDNF were 50ng/ml.Phase contrast microscope observation was done.At the 3rd and 7th day after incubation,the cells were detected by ?-tubulin3 immunofluorescence and Hoechst staining.The length of neuritis was measured,and the numbers of neuron cells and nuclears were determined.At the 1st,3rd,5th,7th and 9th day MTT method was used,and the growth curve was made according to OD results.[Result] The length of axon and positive rate in the experimental groups were superior to those in the control group(P

OBJECTIVETo compare two kinds of method for treating lumbar tuberculosis with psoas abscess, to provide reference for clinical reasonable select of therapy treatment.

METHODSFrom January 2010 to January 2013,42 patients with lumbar tuberculosis combined with psoas abscess with obvious surgical indications were enrolled, including 24 males and 18 females with an average age of (38.5 ± 10.2) years old ranging from 21 to 63 years old. All patients were followed up for 18 to 24 months with an average of 20.9 months. Twenty-two patients underwent posterior vertebral body lesions cleared, bone graft fusion and internal fixation and percutaneous puncture catheter drainage for treatment of psoas major abscess as group A, and twenty patients underwent one-stage extraperitoneal approach to remove abscess, posterior vertebral body lesions cleared, bone graft fusion and internal fixation as group B. The operative time, loss of blood, length of hospital stay, clinical cure rate and other clinical results for the two groups were analyzed and compared.

RESULTSThe loss of blood was (452.3 ± 137.6) ml in group A and (603.5 ± 99.6) ml in group B, there was significant statistical difference (P < 0.05). The time of operation was (193.6 ± 91.2) min in group A and (230.5 ± 56.6) min in group B, there was significant statistical difference (P < 0.05). The time of operation and the loss of blood in group A were obviously less than which in group B. In group A 20 cases were cured and 2 cases relapsed, 19 cases were cured and 1 case relapsed in group B, there was no significant statistical differences between two groups regarding cure rate with chi-square test (χ² = 0.000, P = 1.000). All patients in two groups obtained good clinical curative effect. There were no significant statistical difference between two groups regarding for length of hospital stay with t-test (P > 0.05).

CONCLUSIONLumbar spinal tuberculosis with psoas abscess is not absolute indications for anterior open operation. Compared with the combined anterior and posterior surgical procedure, the percutaneous puncture catheter drainage combined with posterior debridement, interbody fusion and internal fixation can achieve the same clinical effect but less trauma for the patients.

Adult ; Case-Control Studies ; Debridement ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Psoas Abscess ; etiology ; surgery ; Spinal Fusion ; Tuberculosis, Spinal ; complications ; surgery ; Young Adult

Objectives To investigate dynamic pathological features and virus distribution in the liver with a murine severe acute respiratory syndrome(SARS)model injected with murine hepati- tis virus 3(MHV-3)through trachea.As a representative of host genes,mouse fgl2(mfgl2)pro- thrombinase gene expression and its clinical significance were discussed in SARS associated liver dam- ages.Methods The Balb/cJ mice were infected with 100 PFU of MHV-3 through trachea and Balb/ cJ mice injected with saline were served as control.Survival rate,pathological features in organs and liver function were observed.Virus titers in different organs were determined on monolayer of L2 cells by a standard plaque assay.Virus distribution and cellular localization were studied by in situ hy- bridization.Both mfgl2 and fibrin expressions were examined in the liver by in situ hybridization and immunohistochemistry to investigate the role of mfgl2 in the liver impairment.Results Mice infected with MHV-3 through trachea developed multiple organs damages and died within 5 days,while all mice in control group survived with no histopathological changes.Infected liver tissues showed wide- spread cloudy swelling,prominent ballooning degeneration with mild lymphocytic infiltration in the portal area.Dot and zonal hepatocellular necrosis could be found occasionally.The lungs showed typi- cal interstitial pneumonia and hyaline membranes formation.Other histological changes also could be found in other organs examined.MHV-3 virus replication was identified in all organs observed.The liver function was injured,mfgl2 expression were evidenced mainly in the necrosis areas with fibrin deposition around the necrosis areas.Conclusions Pathological changes of the liver in this murine SARS model can mimic the liver impairment characteristics of SARS in human.In addition to the physical damage induced by the virus,the up-regulation of novel gene mfgl2 in the liver in association with fibrin deposition may play a vital role in the development of SARS associated liver damages.

Objective To observe the immunologic effect of lactoferrin on repeated respiratory infection(IRRI) in children.Methods Ninety-eight cases of IRRI were divided into two groups randomly.The control group (48 cases)were treated with routine therapy.The treatment group(50 cases) were treated with lactoferrin based on routine therapy for 2-3 months.T cell subgroup,immunoglobulin and complements were determined before and after treatment.Results Total effective rates in treatment group and control group were 86% and 22.9% respectively.The therapeutic efficacy in treatment group was significantly higher than that in control group(P

The endophytic microorganisms found widely in many kinds of plants mediate various effects to theirs hosts. In this study, seven different dominant endophytes (SDE01 to 07) isolated from a Hy-peraccumulator-Solanum nigrum L. were resistant to Cd2+, and the strain SDE06 survived even in the medium containing 80 mg/L of Cd2+. Bacteria strain SDE06 was identified as Bacillus sp.. The removal of Cd2+ of SDE06 in different conditions were studied. Under the optimal conditions, the incubating time was 36 h, the solution pH 6.0, the temperature was 37?C and the Cd2+ concentration of medium was 20 mg/L, the highest removal rate was up to 80.2% at this condition.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Objective - To analyze the relationship between cobalt level of post shift urine and individual exposure level of ， cobalt and its compounds in cobalt exposed workers and to explore the feasibility of using urine cobalt as a biomarker. Methods - A total of 148 occupational cobalt exposed workers from a new material company were selected as the exposed ， - - group and 44 non occupational cobalt exposed workers from the company were selected as the control group using the typical sampling method. The exposure concentration of time weighted average of cobalt and its compounds in the workplace air of the - two groups was determined by inductively coupled plasma mass spectrometry as the individual exposure level. The cobalt levels - - of pre shift and post shift urinary samples of the two groups were detected by this method. The linear relationship between the - cobalt level of post shift urine and the individual exposure level of cobalt and its compounds in the air of the workplace was Results - 3 analyzed. The individual exposure level of cobalt and its compounds in the exposed group was 1.10 131.71 μg/m with （M） 3 the median of 12.23 μg/m. No cobalt and its compounds were detected in the workplace air in the control group. The cobalt - - levels of pre shift and post shift urines in exposed group were higher than those in the control group at the same time point （M： vs ， vs ， P ） - - 1.54 0.56 μg/L 8.77 0.83 μg/L all <0.01 . The cobalt level of post shift urine was higher than that in pre shift （M： vs ，P ）， urine in the exposed group 8.77 1.54 μg/L <0.01 and it was positively correlated with the individual exposure level （ ，P ） ， of cobalt and its compounds Spearman correlation coefficient=0.86 <0.01 . After common logarithm conversion the linear regression equation of the cobalt level of post shift urine and the common logarithm of individual exposure level of cobalt and （x） ：ŷ x（ ；F ， its compounds in the exposed group was as follows = −0.178 + 0.988 coefficient of determination=0.72 =374.75 P ；t ， P ） Conclusion - <0.01 = - 19.36 <0.01 . There was a linear correlation between cobalt level of post shift urine and occupational cobalt exposure level of cobalt exposed workers. Urinary cobalt can be used as a biomarker of occupational cobalt

To establish kinetic assay method for the analysis of hemolysis and to investigate dynamic hemolytic process of polysorbate 80. The UV-VIS spectrum of heme changes when hemoglobin is released continuously during the hemolytic process. Therefore, dynamic hemolytic curve was determined as a new way to characterize the kinetic process of interaction between polysorbate 80 and red blood cells. The effect of polysorbate 80 on blood cells could be perfectly investigated by the hemolytic dynamics. Dynamic hemolytic parameters of polysorbate 80 were calculated according to the hemolytic curves. The constants of hemolytic rate and maximum hemolytic rate of polysorbate 80 had fine linear relationships at the range of 1-20 mg x mL(-1) and 5-20 mg x mL(-1), respectively. In comparison with the present official method such as macroscopic observation and static spectrophotometric methods, kinetic spectrophotometry has the advantages of real time, on-line determination, sensitive, objective, good reproducibility and 2-dimensional information acquired. Therefore, as a biological technique, kinetic spectrophotometry could be applied to evaluate the quality of polysorbate 80 and to screen other solubilizing excipients.

The inhibitory ratio (1%) of artificial musk on cyclooxygenase-2 (COX-2) was determined by enzyme immunoassay (EIA). The dose-effect relationship between concentrations of artificial musk and 1% was established. It was found that artificial musk had obvious inhibitory action on COX-2. The concentration for 50% of maximum inhibitory effect (IC50) was about 2.26 mg x mL(-1). There was a good relationship between the logarithm concentrations of artificial musk and 1% when the concentrations of artificial musk ranged from 0.31-20.0 mg x mL(-1). The results indicated that this EIA method could be applied to evaluate the anti-inflammatory activity of artificial musk quickly, conveniently, sensitively and exactly. This paper provided a novel method and foundational research for the bioassay of artificial musk.