The aim of the study is to determine the characteristics of the material properties of various structures in the temporomandibular joint (TMJ) and the mandibular bone tissues in nanoscopic scale. Using the atomic force microcopy (AFM) with the nanoindentation method, the nanomechanical properties of 6 discs, condylar cartilages and fossate cartilages in the TMJ, and 6 cortical and cancellous bones in the mandible of 3 normal adult men, were measured and analyzed. Results showed that marked differences were found in elastic properties among the different regions of the disc, articular cartilage in the TMJ. In the distribution of elastic modulus in these various structures, the elastic modulus was higher in the anterior and medial regions and lower in the middle, posterior and lateral regions. Otherwise, elastic modulus of the cortical bone in the mandible was approximately 2 times more than the cancellous bone. Elastic modulus in the buccal bone tissues of the mandible was more distinctly below one of the lingual site. The results suggested that the disc, condylar cartilage and fossate cartilage in the TMJ and the cortical and cancellous bone in the mandible were inhomogeneous with the nanolevel measurement. Different structures or various regions in the same structure were loaded by different local mechanical forces in the nanoscale.

Objective To examine the brain metabolic changes in WD patients receiving copper chelation by us?ing 1H-MRS. Method Thirty-nine patients with WD was randomly divided into four groups: non-brain type group (18 cases), brain type prior-treatment group and short-term treatment group (21 cases), long-term treatment group (20 cases) from short-term treatment group, and 20 healthy volunteers served as a control group. 1H-MRS and MRI were performed on patients on 1.5/MR/MRS system to detect these above-mentioned items before and after treatment. Result The mean of NAA/Cr was significantly lower in the left putamen and head of the caudate nucleus than in the left basal ganglion in the 39 patients with WD. The mean of NAA/Cr and Cho/Cr in the left putamen and basal ganglion was significantly lower in non-brain type group than in control group(P<0.01). The mean of NAA/Cr Cho/Cr and NAA/Cho in the left putamen,head of the caudate nucleus and basal ganglion were significantly lower in brain type group than in control group(P<0.01 or P<0.05). The mean of NAA/Cr in the left putamen was much lower in brain type group than in non-brain type group (P<0.01). The mean of NAA/Cr, Cho/Cr and NAA/Cho of short-term treatment group in the left putamen, head of the caudate nucleus and basal ganglion was not significantly different between brain type group and short-term treatment group(P>0.05). The mean of NAA/Cr and NAA/Cho in the left putamen and basal ganglion was much higher in long-term treatment group than in brain type group(P<0.01 or P<0.05). The mean of Cho/Cr in the left head of caudate nucleus were much higher after treatment compared with prior-treatment group(P<0.05). The mean of NAA/Cr in the left putamen, head of the left caudate nucleus and basal ganglion in all groups was negatively correlated with course of the disease. Conclusion There are significant differences in brain metabolism among different type of WD. The long-term but not short-term copper chelation significantly improves brain metabolism. NAA/Cr may be used as a non-invasive indicator to examine the efficacy of treatment.

OBJECTIVETo explore the application of serum protein pattern models in diagnosis of colorectal cancer (CRC) by proteinchip technology.

METHODSOne hundred and forty-seven serum samples (55 CRC patients and 92 healthy individuals) randomly divided into training set (n = 87, 32 CRC patients and 55 healthy individuals) and test set (n = 60), were subjected for analysis by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). Four top-scored peaks in 5910, 8930, 4476 and 8817 were detected by proteinchip software version 3.0. and were trained by a multi-layer artificial neural network (ANN) with a back propagation algorithm. An artificial neural network classifier had developed for separating CRC from the healthy group. The classifier was then challenged with the test set (60 samples including 23 CRC patients and 37 healthy individuals) to determine the validity and accuracy of the classification system.

RESULTSThe artificial neural network classifier separated the CRC from the healthy samples, with sensitivity of 82.6% and specificity of 91.9%.

CONCLUSIONCombination of SELDI-TOF-MS with the artificial neural network yields significant higher sensitivity and specificity than CEA in the diagnosis of CRC, which should be further studied.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Blood Proteins ; analysis ; Colorectal Neoplasms ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Neural Networks (Computer) ; Protein Array Analysis ; Proteomics ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

To construct the adenoviral vector with co-expressing keratinocyte growth factor (KGF) and enhanced green fluorescent protein (EGFP) for transfection into the mesenchymal stem cells (MSC), the target gene KGF was cloned into the shuttle plasmid with the report gene EGFP, then the recombinant shuttle plasmid was transformed into DH5a bacteria to recombine with backbone vector pAdxsi. Next, the plasmid pAd-EGFP-mKGF was amplified in H293 cells and the viral titer was determined. The MSC were separated and enriched by using bone marrow adherent culture and identified in vitro to observe the efficiency of transfection. The results indicated that the recombinant shuttle plasmid pShuttle-EGFP-mKGF digested with restriction endonucleases was confirmed by two products which length was about 0.6 kb and 5.1 kb, respectively; the recombinant plasmid pAdxsi-EGFP-mKGF digested with restriction endonucleases was confirmed by 7 products; recombinant adenoviral vector Ad-EGFP-mKGF was amplified to titer of 1.6 × 10(10) pfu/ml. At 10 h after transfecting MSC began to express fluorescence at 6 to 8 days later, the fluorescence reached to the peak with infection rate of 92.3, at 28 days the expression of fluorescence was still observed. It is concluded that the recombinant adenoviral vector Ad-EGFP-mKGF is successfully constructed and can transfect MSC effectively and safely.
Adenoviridae ; genetics ; Animals ; Bone Marrow Cells ; cytology ; Fibroblast Growth Factor 7 ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Plasmids ; Transfection

OBJECTIVETo establish serum protein fingerprint model for early diagnosis of pancreatic cancer with surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) and bioinformatics techniques.

METHODSA total of 73 samples were analyzed in this study, including 31 cases of pancreatic cancers, 22 cases of pancreatitis and 20 healthy individuals. Samples were first analyzed by SELDI-TOF-MS and two patterns of differentiation model were constructed with support vector machine arithmetic method.

RESULTSThe pattern 1 model differentiating pancreatic cancer patients from healthy individuals had a specificity and a sensitivity of both 100.0%. The pattern 2 model differentiating pancreatic cancer from pancreatitis had a specificity of 95.5% and a sensitivity of 93.5%.

CONCLUSIONSELDI-TOF-MS technique combined with bioinformatics can facilitate to identify biomarkers for pancreatic cancer.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; Protein Array Analysis ; methods ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Support Vector Machine

Adult ; Aged ; Bone Transplantation ; Female ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Thoracic Vertebrae ; injuries ; surgery

OBJECTIVETo establish the diagnostic model of cerebrospinal protein profile for gliomas by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and bioinformatics.

METHODSSeventy-five samples of cerebrospinal fluid from patients with gliomas, benign brain tumors and mild brain traumas were collected. A total of 50 samples from gliomas and non-brain-tumors were divided into training sets (33 cases including 17 gliomas and 16 non-brain-tumors) and testing sets (17 cases including 5 gliomas and 12 non-brain-tumors). The cerebrospinal proteins bound to H4 chip were detected by SELDI-TOF MS, the profiles of cerebrospinal protein were gained and then analyzed with artificial neural network algorithm (ANN); and the diagnostic model of cerebrospinal protein profiles for differentiating gliomas from non-brain-tumors was established. Forty-seven of cerebrospinal samples of gliomas and benign brain tumors were divided into training sets (31 cases including 13 gliomas and 18 benign brain tumors) and testing sets (16 cases including 9 gliomas and 7 benign brain tumors), the diagnostic model of cerebrospinal protein profiles for differentiating gliomas from benign brain tumors was established based on the same method. The support vector machine (SVM) algorithm was also used for evaluation, both results were very similar, but the result derived from ANN was more stable than that from SVM.

RESULTThe diagnostic model of cerebrospinal protein profiles for differentiating gliomas from non-brain-tumors was established and was challenged with the test set randomly, the sensitivity and specificity were 100% and 91.7%, respectively. The cerebrospinal protein profiling model for differentiating gliomas from benign brain tumors was also developed and was challenged with the test set randomly, the sensitivity and specificity were 88.9%, and 100%, respectively.

CONCLUSIONThe technology of SELDI-TOF MS which combined with analysis tools of bioinformatics is a novel effective method for screening and identifying tumor biomarkers of gliomas and it may provide a new approach for the clinical diagnosis of glioma.

Adult ; Aged ; Algorithms ; Biomarkers, Tumor ; Brain Neoplasms ; cerebrospinal fluid ; diagnosis ; Cerebrospinal Fluid Proteins ; genetics ; Diagnosis, Differential ; Female ; Glioma ; cerebrospinal fluid ; diagnosis ; Humans ; Male ; Meningioma ; cerebrospinal fluid ; diagnosis ; Middle Aged ; Neural Networks (Computer) ; Peptide Mapping ; standards ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The aim of this study was to investigate the effect of mesenchymal stem cells (MSCs) on cell cycle and apoptosis of thymus, spleen and bone marrow cells in mice totally irradiated with sublethal dose, and to explore its mechanisms. BALB/c mice irradiated with 5.5 Gy 60Co gamma-ray were randomly divided into control group and MSC group. Mice in MSC group were infused with 0.4 ml containing 2.5x10(7)/kg of MSCs through tail vein at 1 hour after irradiation. Mice in control group were infused with 0.4 ml normal saline. The cell apoptosis and cell cycle of thymus, spleen and bone marrow cells were detected by flow cytometry at 6, 12, 24 and 72 hours after irradiation and the P53 protein expressions in thymus and bone marrow cells were assayed by immunohistochemistry at 12 hours after irradiation. The results showed that the arrest of cells in G0/G1 and G2/M phase, and decrease of cells in S phase appeared at 6 hours after irradiation, those reached peak respectively at 12 hours in thymus cells, 6 hours in spleen and 24 hours in bone marrow, then the cell counts in G0/G1 phase decreased and the cell counts in S and G2/M phases increased. At 72 hours the cell counts in G0/G1 phase were less than the normal level and the cell counts in S phase were more than the normal level. The above changes of cell cycle in thymus and spleen were more rapid in spleen and more obvious in amplitude than that in bone marrow, the change of cell cycle in MSC group was more rapid and obvious than those in control group. After irradiation the apoptosis cells increased from 6 hours, reached the highest level at 12 hours and decreased to the normal level gradually after 24 hours in two groups; the apoptosis rates in spleen and thymus cells were higher than that in bone marrow cells. In comparison with the control group, the apoptosis rate in thymus cells at 12 hours, in spleen cells at 12 and 24 hours, and in bone marrow cells at 24 hours were fewer in MSC group. The cells expressing P53 protein in control group were more than that in MSC group. It is concluded that the MSCs accelerate the running of cell cycle in these hematopoietic tissue cells of irradiated mice, reduce the cell apoptosis and promote the recovery from injuries in hematopietic and immunological organs, thus protect the irradiated mice at early stage.
Animals ; Apoptosis ; physiology ; Bone Marrow Cells ; pathology ; Cell Cycle ; Female ; Mesenchymal Stem Cell Transplantation ; Mice ; Mice, Inbred BALB C ; Radiation Injuries, Experimental ; pathology ; therapy ; Random Allocation ; Spleen ; pathology ; Thymus Gland ; pathology ; Whole-Body Irradiation

To explore hemorrhage risk and the clinical significance of abnormal change of prothrombin time (PT), activated partial thromboplastin time (APTT), plasma fibrinogen (FIB), plasma thrombin time (TT) and d-dimer (D-D) in de novo acute leukemia (except for APL), the different bleeding manifestations of 114 cases of de novo acute leukemia with different coagulation indexes were analyzed retrospectively. The correlation between these blood coagulation indexes and the possible correlative clinical characteristics were analysed, including age, sex, type of acute leukemia, initial white blood cell(WBC) and platelet(Plt) count, the proportion of blast cells in bone marrow and cytogenetic abnormality of patients at diagnosis. The results indicated that the incidence of abnormal blood coagulation was as high as 78.1% for de novo AL patients. These patients with 5 normal blood coagulation indexes may have mild bleeding manifestation, but the more abnormal indexes, the more severe bleeding. Both PT and D-D were sensitive indexes for diagnosis of level II bleeding. Incidence of abnormal blood coagulation significantly correlates with the proportion of blast cells in bone marrow (χ(2) = 4.184, OR = 1.021, P < 0.05) and more with D-D (P < 0.01), while age, sex, type of AL, WBC count, Plt count and abnormality of cytogenetics did not correlate with abnormal blood coagulation. It is concluded that the coagulation and fibrinolysis are abnormal in most patients with de novo acute leukemia. More abnormal indexes indicate more severe bleeding, and both PT and D-D are sensitive indexes for diagnosis of level II bleeding. Higher proportion of blast cells in bone marrow predicts higher incidence of abnormal blood clotting. Acute leukemia with elderly age, high white blood cell count and adverse cytogenetics do not predict severer abnormal blood clotting. Detection of PT, APTT, TT, FIB, and D-D may help to judge whether the patients are in a state of hypercoagulability or disseminated intravenous coagulation, which will provide experiment evidences for early intervention and medication.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Coagulation ; Blood Coagulation Tests ; Child ; Female ; Fibrin Fibrinogen Degradation Products ; Hemorrhage ; pathology ; Humans ; Leukemia ; blood ; pathology ; Leukocyte Count ; Male ; Middle Aged ; Platelet Count ; Prothrombin Time ; Retrospective Studies ; Thrombin Time ; Young Adult

OBJECTIVETo observe the therapeutic effect and major complications of haploidentical nonmyeloablative allogeneic peripheral blood stem cell transplantation (NST) for refractory or relapsed leukemia.

METHODSThe results of 30 patients, including 14 cases of acute myeloid leukemia (AML), 11 cases of acute lymphoblastic leukemia (ALL), 5 case of chronic myelogenous leukemia (CML) (accelerated and blastic phase) with refractory or relapsed leukemia (RF/RL) who underwent haploidentical NST from August 2000 to April 2009 were analyzed. The conditioning regimen consisted of fludarabine (flu), antithymocyte globulin (ATG), cyclophosphamide (CTX), total body irradiation (TBI) and cytarabine (Ara-C) or myleran (Bu). Graft-versus-host disease (GVHD) prevention programmes consisted of Cyclosporine (CsA), mycophenolate mofetil (MMF), CD25 monoclonal antibody combined with mesenchymal stem cells (MSC).

RESULTSTwenty six cases of patients were full donor engraftment and 4 cases mixed chimerism into full donor chimerism. The average duration of neutrophil >0.5×10⁸/L after NST was 11 (9-16) days, and platelet >20×10⁸/L 17 (12-60) days. Upon follow-up of 16 to 120 months, 12-month transplant-related mortality (TRM) was 46.7%, acute Ⅱ-Ⅳgraft-versus-host disease (aGVHD) incidence was 40.0%. The probability of 3-year disease relapse, EFS and overall survival (OS) rates were 16.7%, 46.2% and 50.0% respectively.

CONCLUSIONHaploidentical NST could improve OS and EFS of refractory or relapsed leukemia and reducce TRM to some extent.

Adolescent ; Adult ; Child ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia ; therapy ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Survival Rate ; Treatment Outcome ; Young Adult