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 observe the effect of sirolimus-based immunosuppression administered on heart transplant recipients with chronic renal dysfunction.

METHODSFrom June 2004 to December 2008, standard calcineurin inhibitors (CNI)-based immunosuppressive regimen was changed to reduced-dose CNI plus sirolimus due to CNI-related chronic renal dysfunction in 20 out of 138 cardiac transplant recipients at Fuwai Hospital. The standard immunosuppressive regimen included steroid, CNI (cyclosporine or tacrolimus), and mycophenolate mofetil or azathioprine. Sirolimus was started at 0.75 - 1.50 mg/d with titration to achieve levels of 5 - 15 µg/L, and CNI dose was reduced gradually to 1/2-2/3 of the baseline level. Patients were followed for changes in renal function, lipid level and clinical side effects related to immunosuppressive therapy. Endomyocardial biopsy (EMB) was performed routinely at 3 weeks, 3, 6 and 12 months after transplantation. EMB was also performed at 3 months after regimen change within 1 year post-transplantation or when rejections were suspected in patients beyond 1 year post-transplantation. Echocardiography was performed for monitoring purpose.

RESULTSThe mean follow-up after regimen change was (7.9 ± 6.3) months. Final sirolimus dose was (0.89 ± 0.22) mg/d and blood drug level was (7.6 ± 3.8)µg/L. Cyclosporine dose was reduced from (191.7 ± 60.0) mg/d to (123.6 ± 34.8) mg/d, with blood drug concentration reduced from (175.5 ± 58.0) µg/L to (111.9 ± 56.0) µg/L in 18 patients (P < 0.01). Tacrolimus average dose was reduced from 4.25 mg/d to 3.00 mg/d, with blood drug concentration reduced from 13.5 µg/L to 10.5 µg/L in 2 patients. Serum creatinine level fell from (160.4 ± 25.5) µmol/L to (134.4 ± 26.8) µmol/L (P < 0.01) and urea nitrogen fell from (13.8 ± 4.7) µmol/L to (10.4 ± 3.0) µmol/L (P < 0.01) at one month after regimen change. Twenty two EMBs were performed in 11 patients within 1 year post-transplant, there were 4 episodes of acute rejected (ISHLT grade 2). Twenty patients are all alive and cardiac function was normal. The most common side effect was hyperlipidemia, and triglycerides, total cholesterol and low density lipoprotein levels were significantly increased at 1 month post regimen change (P < 0.05 or P < 0.01). Leukocyte, hemoglobin and platelet as well as liver function remained unchanged at 1 month post regimen change (all P > 0.05).

CONCLUSIONOur results show that change from CNI-based immunosuppressive regimen to reduced-dose CNI plus sirolimus is an effective and safe approach for the management of patients with CNI-related chronic renal dysfunction, leading to an improvement in renal function without compromise in anti-rejection efficacy and with tolerable side effects.

Calcineurin Inhibitors ; Female ; Heart Transplantation ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Kidney Failure, Chronic ; drug therapy ; Male ; Middle Aged ; Retrospective Studies ; Sirolimus ; therapeutic use

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

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

The aim of this study was to investigate effects of IAT and MAT chemotherapeutic regimens treating patients with refractory and relapsed acute myeloid leukemia (AML). 99 patients with refractory and relapsed AML received IAT regimen or MAT regimen as study objects were retrospectively analyzed (56 patients with refractory AML and 43 patients with relapsed AML). Among of them, 28 patients were treated with IAT regimen, and 71 patients received with MAT regimen. The results showed that in 2 groups mentioned above the OR was 65.7%, CR was 49.5%, PR was 16.2%; in IAT group the OR was 64.3%, CR was 46.4%; in MAT group the OR was 66.2%, CR was 50.7%, no statistical difference was found between these 2 groups; The 2 years overall survival was 25% in IAT group and 15.5% in MAT group. Serious infection in IAT and MAT regime groups was 25% and 9.9%, respectively. It is concluded that both IAT and MAT regimens are effective methods for inducing CR in patients with refractory of relapsed AML. IAT and MAT regimens can be used in treatment of the refractory or relapsed MAL patients who were not respond to other regimen.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Child ; Cytarabine ; administration & dosage ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; etiology ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Vidarabine ; administration & dosage ; Young Adult

This study was aimed to investigate the effect of recombinant human granulocyte colony-stimulating factor(G-CSF) on murine thymocyte emigration and cell cycle alteration after a sublethal dose of gamma-irradiation. Female BALB/c mice were given 6.0 Gy γ-ray total body irradiation and then randomly divided into G-CSF and control groups. Mice in the G-CSF group were injected recombinant human G-CSF 100 µg/(kg·d) subcutaneously once daily for 14 consecutive days and mice in the control group were given the same volume of phosphate buffered solution. Thymocyte cycle alteration and the proportion of apoptosis cells were detected by flow cytometry within 72 hours after irradiation. Real-time PCR was used for detection and quantitation of murine T cell receptor rearrangement excision circles (sjTREC) of the thymic cells at 30 and 60 day after the irradiation. The results showed that at 6 hour after irradiation G-CSF could significantly increase the thymic cells in G(0)/G(1) phase, G-CSF vs control: (82.0 ± 5.0)% vs (75.9 ± 2.8)% (p < 0.05), and decrease the thymic cells in S phase, G-CSF vs control: (10.2 ± 4.8)% vs (15.7 ± 2.3)% (p < 0.05), but G-CSF seemed have no evident effects on the percentage of thymic cells in G(2)/M phase. G-CSF could also protect thymocytes from apoptosis at 6 hour and 12 hour after irradiation the percentages of apoptosis cells in G-CSF group were (11.5 ± 2.4)% and (15.5 ± 3.3)%, respectively, which were significantly lower than that of the control group (16.5 ± 2.2)% and (22.6 ± 0.7)%, respectively (p < 0.05). The sjTREC copy amount was conspicuously higher in G-CSF group than that in the control at 30 day after irradiation (p < 0.01), but the preponderance disappeared 60 days later. It is concluded that G-CSF has a positive effect on the thymic cell cycle alteration to protect thymocytes from apoptosis and enhance the recent thymocyte emigration, which may contribute to the central immune reconstitution after irradiation.
Animals ; Cell Cycle ; drug effects ; radiation effects ; Female ; Gamma Rays ; adverse effects ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; pharmacology ; Thymocytes ; drug effects ; radiation effects