Objective:To fabricate and to study the surface morphology and biological safety of a novel coating on microarc-oxidized titanium.Methods:The novel functional coating was fabricated by cross-linking the double-layer nanoparticles loading rhBMP-2 and SDF-1 with gelatin on microarc-oxidation coating on titanium implant surface.The surface topography was observed and optimized,and the biological safety of the novel coating was primarily evaluated by cell toxicity test,oral mucosa stimulation test and hemolysis test in vitro.Results:The novel functional coating possesses excellent morphology.The coating showed the cytotoxicity of score 1 and no mucous membrane irritation,the hemolytic rate of the coating was 4.6％.Conclusion:The coating possesses good morphology and biological safety.

Objective To investigate the complications related factors in infants with mycoplasma pneumonia pulmonary(MPP). Methods According to the condition of pulmonary complications, 105 cases of infants MPP were divided into pulmonary complication group and no pulmonary complication group with 72 cases and 33 cases respectively,and the general related factors and disease related factors of two groups were analyzed. Results The incidence rate of pulmonary complication was 68.6% (72/105) in infants MPP, and the main involved extra-pulmonary systems were digestive system (54.2%), cardiovascular system (44.4%) and blood system(33.3%). Among 20 factors associated with pulmonary complications of MPP, age, feeding method (including artificial, mixing and milk three classification), season of onset, fever days, the titer of mycoplasma pneumonia (MP)-IgM, C-reaction protein, erythrocyte sedimentation rate and the initial time of using macrolides had significant differences between two groups (P<0.05). Multivariate Logistic regression analysis showed that age≥2 years, fever days≥7 d, titer of MP-IgM≥1∶160, increased C-reaction protein levels and accelerated erythrocyte sedimentation rate were the risk factors for pulmonary complications of infants MPP, while breastfeeding and using macrolides within 7 d were the protective factors. Conclusions The incidence rate of pulmonary complications in infants MPP is high, which can affect multiple systems. For children with older age, longer thermal process, higher titer of MP-IgM, and increased C-reaction protein , accelerated erythrocyte sedimentation rate and more past medical history, more attention should be paid for their higher pulmonary complications incidence.

Objective:To construct double-layered controlled release system containing SDF-1 and rhBMP-2 molecules and to study the release profile of the system in vitro.Methods:The polylactic acid/chitosan(PLA/CS)nanoparticles were prepared with “emulsification-solution evaporation method”,the preparation parameters were determined by orthogonal test design.The particle size was observed by nanoparticle size analyzer,the morphology of the nanoparticles was observed with electron microscope.Then rhBMP-2 and SDF-1 were loaded into the nanoparticles in the process of emulsification,the loading efficiency and encapsulation efficiency were calculated and in vitro release was observed.Results:The double-layer nanoparticles showed spherical geometry,smooth surface and complete separation. The average particle size of the nanoparticles was (542.33 ±14.38)nm;The drug loading and incorporation efficiency of rhBMP-2 were (82.41 ±1.05)% and (24.67 ±0.43)ng/mg,those of rhBMP-2 were (75.58 ±0.84)% and (22.63 ±0.41)ng/mg,respectively. The release time of the drug from the system sustained over at least 30 days,the release profile of both drugs showed “biphasic release”. The cumulative release rate of SDF-1 and rhBMP-2 was 72.85% and 91.01% in 30 days respectively.Conclusion:The SDF-1 and rh-BMP-2 loaded PLA/CS nanoparticles have excellent morphology,high entrapment and good sustained-release in vitro.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. However, emergence of drug resistance limits its potential use. Plumbagin is a natural quinonoid compound isolated from plant. In this study, induced apoptosis effect of the combined treatment with plumbagin and TRAIL on human melanoma A375 cell line was examined and possible mechanism was investigated. The cells were divided into four groups: control group, plumbagin group (plumbagin, 5 or 10 mumol/L), TRAIL group (TRAIL, 30 ng/mL) and plumbagin+TRAIL group (combined treatment group). The apoptosis, and the expression of DR4 and DR5 were detected by flow cytometry. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that the apoptosis rate was 8.3% in TRAIL group, 10.35%-16.94% in plumbagin group and 52.39%-65.39% in combined treatment group, respectively, with the difference being significant between combined treatment group and plumbagin or TRAIL group (P<0.05 for each). Moreover, plumbagin alone could markedly up-regulate DR5 mRNA and protein expression, and slightly increase DR4 mRNA and protein expression. Treatment of human melanoma A375 cells with plumbagin resulted in the activation of Caspase-3, but not Caspase-8. These results suggest that plumbagin might be useful for TRAIL-based treatment for melanoma.

Objective To detect the role of differentially expressed microRNA (miRNA) in human cholangiocarcinoma and explore their effects on apoptosis and invasiveness of cholangiocarcinoma.Methods The differential expression of miRNA in 3 cholangiocarcinoma patients was detected by miRNA array.The expressions of miR-200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues and normal bile duct tissues were detected by real-time PCR.After transfection with miR-200b mimic,apoptosis and invasiveness of human cholangiocarcinoma cell line QBC939 was evaluated by Annexin-V-FITC dyeing and Transwell assay.Results Comparad with normal bile duct tissues,the number of differential miRNAs in cholangiocarcinoma was 21,including 15 up regulated and 6 down regulated.The expressions of miR200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues were significantly lower than levels in normal bile duct tissues.The invasive ability of QBC939 was decreased after miR-200b mimic transfection.The apoptosis cell number of QBC939 was increased after miR-200b mimic transfection.Conclusion These results indicate that the expression of miRNA is different between cholangiocarcinoma and normal bile duct tissues.Moreover,miR-200a,miR-200b,miR-200c,and miR-141 are likely involved in the invasion and metastasis of cholangiocarcinoma and have potential as a diagnostic and prognostic marker.

Objective·To prepare chitosan-gelatin porous microspheres by high voltage electrostatic method combined with freeze-drying and ionic cross-linking method and investigate the factors that influence the formation of porous medium.Methods·Porous chitosan microspheres and chitosangelatin porous microspheres were prepared using high voltage electrostatic method combined with freeze-drying and ionic cross-linking method,with sodium tripolyphosphate (STPP) as crosslinking agent.Factors that affect the porous structure and pore size of porous microspheres were compared,such as different chitosan-gelatin ratio,freezing temperature,curing time with saturated STPP in 85％ ethanol solution.The morphology,surface and internal structure,particle size of the porous chitosan microspheres and chitosan-gelatin porous microspheres were observe by using light microscope,scanning electron microscope and hematoxylin-eosin staining.Results·Microspheres prepared by freeze-drying with an electrostatic and ionic cross-linking method have open,interconnected and highly macroporous,with good spherical surface.Saturated STPP ethanol solution (85％ ethanol) was chosen as the crosslinking agent to prevent destruction of the porous structure.The order of freeze and crosslinking,cross-linking time and the second freezing temperature,can influence the pore size of porous microspheres.Gelatin and chitosan can form polyelectrolyte complexes,and can also be used as porogen in porous structure.Conclusion·The preparation of porous chitosan-gelatin microspheres via this method has a large pore size (diameter 100-200 μm),suitable for cell growth and the migration.

Objective To investigate the operation key points, instrument improvement and shortterm effects in total en bloc spondylectomy (TES) via a single posterior approach for thoracic and lumbar tumors. Methods A series of modified instruments have been designed for the TES, including threadwire saw (T-saw) with a diameter of 0.81 mm, director and clamping for the saw, L shape and furcation osteotomes.The corpectomy of original TES which was defined as "one step dissection" from anteriorly to posteriorly, was modified into "two step dissection" which means that corpectomy was performed with saw cutting anteriorposteriorly and the L shape cutting posterior-anteriorly. In the cases with difficulty in pediculotomy using a T-saw, furcation osteotome was used for pediculotomy. Ten patients with thoracic or lumbar tumors were treated with the modified TES. There were 1 case of bone giant cell tumor, 1 case of bone neurilemmoma and 8 cases of metastatic tumors. All patients suffered moderate-severe pain and neurological deficit. Results The average follow-up period was 8.1(3.3-18.1) months. The average operating time was 7.8 h(6.0-10.3 h),and average blood loss was 2100 ml (1200-3500 ml). No disruption of dural mater, the leakage of cerebrospinal fluid, iatrogenic spinal cord injury and major vessel damage occurred. Two patients who underwent pleura disruption happened during the operation were treated with intrathoracic drain remedy. Among 7 cases with thoracic tumors, significant improvement in neurological function were achieved in 5 patients with the improvement of one grade in ASIA classification, while no change was found in 2 cases. In 3 cases with lumbar tumor, lumbar nerve root pain relieved and the muscle strength had recovered to grade 4 at least postoperatively. Conclusion Significant improvement has been achieved in the maneuverability and safety of the modified surgical techniques in TES with a single posterior approach for thoracic and lumbar tumors.

BACKGROUND:Pain is the main clinical manifestation for ankylosing spondylitis. At present, nonsteroid anti-inflammatory drugs are oral y taken, but the effects are limited and toxic and side effects are more. Thus, there is no effective scheme for treatment of pain induced by ankylosing spondylitis.
OBJECTIVE:To investigate the correlation between postoperative joint pain al eviation and al ogeneic blood transfusion, and the mechanisms.
METHODS:We retrospectively analyzed clinical data of 88 ankylosing spondylitis patients combined with kyphosis who received only one section of osteotomy surgery merging hip joint pain. We compared the visual analog scale score of hip joint and detected the variation of leucocytes, lymphocytes and immunoglobulin concentrations before and after the operation in the groups of fresh al ogeneic whole blood transfusion, autologous whole blood transfusion, and mixed transfusion of al ogeneic and autologous whole blood. Flow cytometry was used to analyze the number and ratio of peripheral blood Th17 cells and Treg cells which were both highly associated with autoimmune diseases.
RESULTS AND CONCLUSION:The symptom of hip arthralgia obviously improved in both groups transfused by fresh al ogeneic whole blood or al ogeneic-autologous mixed whole blood. However, no obvious variation was detected in leucocytes, lymphocytes and immunoglobin concentration. However, flow cytometry results indicated that Th17/Treg proportion associated with autoimmune diseases was increased remarkably in peripheral blood of ankylosing spondylitis patients. Results suggested that al ogeneic whole blood transfusion can al eviate patients’ joint pain by correcting the imbalance of Th17/Treg which may improve their immune state.

OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.

BACKGROUND: Hepatocyte/polymer interface with good biocompatibility is the key factor in bioreactor design andconstruction, however, bioreactor used in the clinical practice currently is not an ideal one.OBJECTIVE: To establish human hepatocyte compatible polypropylene interface and to lay a foundation for establishingbioartificial liver reactor with polypropylene hollow fiber.DESIGN, TIME AND SETTING: The comparative observation, cell compatibility experiment was performed betweenFebruary and October 2003 at Shanghai Jiao Tong University, Shanghai, China.MATERIALS: Polypropylene Photochemical graft polymerization modification technique was used to graft hydrophilicacrylamide groups on the surface of polypropylene membrane by chemical bonds to form modified polypropylenemembrane.METHODS: L02 human hepatoeytes were seeded on polypropylene membrane, modified polypropylene membrane andpolystyrene membrane, and polystyrene membrane was used as normal control.MAIN OUTCOME MEASURES: Static water contact angle of polypropylene membrane before and after graftmodification; morphology, adherent rate and proliferation activity of L02 human hepatocytes on different material surfaces.RESULTS: Static water contact angle after polypropylene membrane graft modification was smaller than that before graftmodification (P < 0.05). The adherent rate of L02 human hepatocytes on the surface of modified polypropylene membranewas 0, and the proliferation activity of them, which grew as spherical aggregates, was markedly higher than that of cells onpolystyrene membrane and polypropylene membrane without graft modification.CONCLUSION: Grafting polyacrylamide on the surface of polypropylene can establish good interface of L02 humanhepatocytes/polypropylene and form hepatocyte spherical aggregates through simple static culture.