BACKGROUND:Different methods to remove immunogenicity have different effects on the spatial microstructure of antigen-extracted heterologous bone.
OBJECTIVE:To compare the microstructure of the antigen-extracted heterologous bone via different methods to provide experimental data for tissue engineering industrialization.
METHODS:Fresh cancellous bones extracted from adult sheep vertebrae were prepared into cylinders. Their long axis direction was the same with orientation of the trabeculae. After vibration washing and different-frequency ultrasound rinsing, the cylinder samples were randomly divided into three groups:in physical calcined group, the samples were defatted, decellularized and deproteinized sequential y using methanol/chloroform and hydrogen peroxide, then bathed in sodium pyrophosphate and directly calcined at 1 000 ℃ for 3 hours;in chemical group, the samples were defatted, decellularized and deproteinized sequential y using methanol/chloroform and hydrogen peroxide;in control group, the samples were dried natural y at room temperature. Microstructure of the samples in each group was analyzed and compared through determination of porosity, scanning electron microscopy observation, X-ray diffraction analysis, X-ray atomic spectroscopy elemental analysis microscopic spatial structure.
RESULTS AND CONCLUSION:The physical calcined and chemical groups maintained natural network pore system to different extents. The size of the large pore was 50-600μm and that of the smal one was about 2μm. The porosity was 55%to 70%. Hydroxyapatite was the main component of the physical calcined group which was determined by X-ray diffraction, and a smal amount of theβ-Tricalcium phosphate was also determined. In the chemical group, the main component was only hydroxyapatite. The three-dimensional spatial structures of the deproteinized cancellous bones were not damaged greatly, and they had a natural pore network system. Antigen component of xenogeneic cancellous bone can be more thoroughly removed by physical calcination step. The scaffold material made by antigen-extracted heterologous bone may satisfy the demands for bone tissue-engineering scaffolds.

Objective To discuss the mode of onset,clinical characteristics,treatment and prognosis of children with granulocyte sarcoma (GS),in order to provide guidance for early diagnosis and effective treatment of GS.Methods Six cases of children with GS diagnosed at the Department of Hematology,Children's Hospital Affiliated to Soochow University between June 2009 and June 2014 were analyzed,the data including the mode of onset,clinical manifestation,diagnosis,treatment and outcome.Results There were 2 cases with a painless mass onset (1 case was 2 years old,characterized by right waist mass,about 10 cm × 5 cm;the other case was 6 years old,characterized by axillary lump,about 2 cm × 3 cm),and both of them received surgical removal of the tumor,then the postoperative tumor was examined by pathologic and immunohistochemical method,and at last the primary granulocyte sarcoma was diagnosed.The third case was a 7 years old girl,she was onset characterized by scalp lump,about 2 cm × 3 cm,and was diagnosed by the pathologic and immunohistochemical method,and changes in hematological system appeared a month later and acute myeloid leukemia(AML) was confirmed by bone marrow examination.The onset ages of other 3 cases were in 10 months,1 year and 7 months,13 years and 3 months old respectively,characterized by scalp lump (about 2 cm × 3 cm),spinal canal tumor (about 1.0 cm × 1.5 cm),intracranial tumors (6.0 cm × 4.9 cm),with AML occurring at the same time,which was confirmed by surgical pathology,immunohistochemistry and bone marrow cell morphology,immune classification,chromosome,and fusion gene diagnosis.Four cases were hematopoietic malignancies by pathology,2 cases of then belonging to small round cell tumor.The immune pathology showed 5 cases of myeloperoxidase positive,CD68-positive,3 cases of CD43-positive,CD123-positive.All children CD3,CD20 levels in all children were negative.Four cases underwent surgery combined with chemotherapy,other 2 cases received surgery and then gave up treatment,1 case discontinued follow-up 3 months later,and the other case died of intracranial hemorrhage after 3 months,which induced by thrombocytopenia.The treated 4 cases were followed up 3 to 58 months,and all had disease-free survival.Conclusions Children with GS have low incidence and non-specific diagnostic criteria,its diagnosis depends on immune pathology,and the treatment is mainly in accordance with AML program for high-dose chemotherapy.The systematic chemotherapy helps to prolong overall survival;at the same time,the hematopoietic stem cell transplantation with bone marrow may help to improve the prognosis.

Objective To investigate the clinical effect of anterior internal fixation plus vacuum sealing drainage in the treatment of ulnar and radial fractures of Gustilo type Ⅲ.Methods Twenty-eight patients with open ulnar and radial fracture of Gustilo type Ⅲ were managed from April 2007 to March 2014,and were divided into four groups(n =4).Group A were managed with external fixator and conventional changing dressings.Group B were managed with internal fixation and vacuum sealing drainage.Group C were managed with external fixator and vacuum sealing drainage.Group D were managed with internal fixation and conventional changing dressings.Result Twenty-eight cases were adopted telephone follow-up for 6 to 27 months.The soft tissue recovery time of each group respectively was (20.5 ± 2.37) days,(14.7 ±2.16) days,(15.6 ±2.17) days and(19.7 ±2.18) days.The hospital stay of each group respectively was (9.7 ± 2.54) weeks,(4.7 ± 1.46) weeks,(5.2 ± 2.34) weeks and 8.6 ± 2.16) weeks.The fracture healing time of each group respectively was (19.6 ± 2.74) weeks,(13.1±1.84) weeks,(18.1 ±2.54) weeks and (14.7 ± 1.74) weeks.There was significant difference of these data between the two groups(P ＜ 0.05).Conclusions Anterior internal fixation plus vacuum sealing drainage is a better way to treat ulnar and radial fractures of Gustilo type Ⅲ.The technique has short period,less complications,less painful and less expense.

BACKGROUND: Effects of embryonic stem cell-derived hepatocyte-like cell transplantation on oncogenicity of differentiated hepatocyte-like cells and biochemical metabolism of liver should be further studied.OBJECTIVE: To evaluate the therapeutic efficacy of embryonic stem cell-derived hepatocyte-like cell transplantation on the acute liver failure.DESIGN: Randomized controlled study.SETTING: the First Affiliated Hospital of Sun Yat-sen University.MATERIALS: This study was performed at the Central Laboratory, the First Affiliated Hospital of Sun Yat-sen University from January 2005 to February 2006. D3-ES cells extracted from the mice which underwent transfection of green fluorescent protein were graciously presented by professor Huang, Ophthalmology Center of Sun Yat-sen University. Forty 6-week-old D3-129 mice of clean grade and irrespective of gender were provided by Experimental Animal Center of Sun Yat-sen University [certification: SCXK (yue) 2004-0011]. The experimentzl animals were disposed according to ethical criteria.Transforming growth factor, basic fibroblast growth factor, and hepatocyte growth factor were provided by Gibco BRL Company, USA.METHODS: Transforming growth factor, basic fibroblast growth factor, and hepatocyte growth factor were combined to differentiate D3-ES cells into hepatic cells. Cell suspension was poured into liver capsule of 20 mice with 2.0×10(6)cells per mouse. Another 20 mice that determined as the controls were injected with saline. Twenty-four hours later, intraperitoneal injection of 5 μL/20 g carbon tetrachloride was used to induce acute liver failure and to observe quality of life and mean survival time. Twenty-four hours after acute liver failure, vena cava posterior blood was drawn to detect total bilirubin,glutamate-pyruvate transaminase, albumin, blood glucose, pro-time prothrombin time, and other hepatic functional parameters. By scarification, hepatic samples were obtained to evaluate oncogenesis condition, and then HE staining and immunohistochemistry were adopted to detect growth of transplanted cells and albumin expression.MAIN OUTCOME MEASURES: Quality of life, average survival time, hepatic functional parameters, growth of transplanted cells, and oncogenesis condition.RESULTS: Quality of life and average survival time: After the onset of acute liver failure, mice in the control group had incoordination and other symptoms of central nervous system. In addition, 14 mice in the control group and 8 in the transplantation group had abdominal dropsy. Average survival time in the control group was significantly shorter than that in the transplantation group (23, 62 hours, P<0.05). Hepatic functional parameters: Levels of total bilirubin and glutamate-pyruvate transaminase in the control and transplantation groups were higher than those before modeling; levels of albumin and blood glucose were lower than those before modeling; pro-time prothrombin time was significantly longer than that before modeling(P<0.01). Furthermore, levels of total bilirubin and glutamate-pyruvate transaminase in the transplantation group were lower than those in the control group; blood glucose in the transplantation group was higher than that in the control group, and pro-time prothrombin time in the transplantation group was significantly shorter than that in the control group (P<0.05). Growth of transplanted cells and oncogenesis condition: Pathological section demonstrated that structure of liver tissue was not changed remarkably, and tumor was not formed. Moreover, transplanted cells and hepatocyte-like cell were well arranged and combined to express albumin.CONCLUSION: Embryonic stem cell-derived hepatocyte-like cell transplantation can improve quality of life, prolong survival time of model mice with acute liver failure; additionally, transplanted cells may well support biochemical metabolism of liver tissue.

OBJECTIVE To investigate the clinical characteristics,risk factors and preventive measures for nosocomial pneumonia in patients with post-hepatitis liver cirrhosis.METHODS A prospective and retrospective study was carried out to investigate the clinical data of 495 patients with post-hepatitis liver cirrhosis in Department of Infectious Diseases during Jan 1,2005 to Dec 31,2007.RESULTS The incidence rate of the nosocomial pneumonia in post-hepatitis liver cirrhosis patients was 13.50 %.The death rate was 25.40 %,which was obviously higher than 6.8% of patients without no nosocomial infection(?2=23.77,P

Objective To establish an expression system of TCR?9/?2-Fc protein by baculovirus vector expression system and identify biological function of expressed TCR?9/?2-Fc protein.Methods ?9Fc and ?2(OT3)Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph.The recombinant plasmid pBACp10ph-?9/?2(OT3)-Fc and the baculovirus DNA were co-transfected into sf9 cells.The expression position of TCR?9/?2(OT3)-Fc was identified by Western blot and the expression efficiency of ?9Fc and ?2(OT3)Fc was tested by flow cytometry(FCM).Furthermore,the binding activity of TCR?9/?2(OT3)-Fc protein with SKOV3 cells and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance(SPR).Results The recombinant vector pBACp10ph-?9/?2(OT3)-Fc was constructed and TCR?9/?2(OT3)-Fc protein was expressed in sf9 cells.However,the expression efficiency of ?9Fc and ?2(OT3)Fc was quite different.It was proved that purified TCR?9/?2(OT3)-Fc protein can bind with SKOV3 cell and MNS protein.Conclusion TCR?9/?2-Fc protein is successfully expressed in baculovirus vector expression system and TCR?9/?2-Fc protein can simulate the binding activity of TCR in vitro.

Objective To establish an expression system of TCRγ9/δ2-Fc protein by baculovirus vector ex-pression system and identify biological function of expressed TCRγ9/δ2-Fc protein. Methods γ9Fc and 82 (OT3) Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph. The recombinant plasmid pBACp10ph-γ9/δ2(OT3)-Fc and the baculovirus DNA were co-transfected into st9 cells. The expression position of TCRγ9/δ2 (OT3)-Fc was identified by Western blot and the expression efficiency of γ9Fc and δ2 (OT3) Fc was tested by flow cytometry (FCM). Furthermore, the binding activity of TCRγ9/δ2 (OT3)-Fc protein with SKOV3 ceils and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance (SPR). Results The recombinant vector pBACp10ph-γ9/δ2(OT3)-Fc was constructed and TCRγ9/δ2(OT3)-Fc protein was expressed in sf9 ceils. However, the expression efficiency of γ9Fc and 82 (0T3) Fc was quite differ-ent. It was proved that purified TCRγ9/δ2 (OT3)-Fc protein can bind with SKOV3 cell and MNS protein. Conclu-sion TCRγ9/δ2-Fc protein is successfully expressed in baculovirus vector expression system and TCRγ9/δ2-Fc protein can simulate the binding activity of TCR in vitro.

To analyze the reasons of injuries during surgical practices and explore the hazards and the corresponding preventive measures. The surgical interns don't receive enough training of injury prevention. They don't pay enough attention to the occupational prevention and are not skilled in operation, which causes high rate of sharp edged instrument injuries. Therefore, a perfect plan of education, training and treatment must be made to reduce the occupational injuries and blood sourced diseases.

Dendritic Cells ; pathology ; Humans ; Leukemia, Myeloid ; pathology ; Plasmacytoma ; pathology

Objective To observe the of changes of cerebral blood flow and electroencephalography in chronic cerebral circulation insufficiency (CCCI) treated with fastigial nucleus stimulation (FNS) and hyperbaric oxygen (HBO). Methods 144 cases of CCCI were divided into 4 groups: 36 cases were treated with FNS and HBO, 36 cases with FNS, 36 cases with HBO, 36 cases without any treatment as control group. The blood velocity of anterior, middle, posterior cerebral arteries, vertebral artery and basilar artery were measured with transcranial Doppler (TCD) and the brain waves （α, β, δ, θ） were recorded with electroencephalography (EEG) before and after the treatment. Results Compared with the control, the brain blood velocity and α wave increased in all the treatment groups, especially in the HBO+FNS group, while β, δ, θ waves decreased (P<0.05). Conclusion FNS and HBO can increase cerebral blood flow and improve the cerebral function respectively.