Objective To study the adaptation process and extent of changes in articular cartilage in the canine knee joint to the intensified training, in order to elucidate the influence of different movement training on remodeling and reconstruction of articular cartilage. Methods Twenty adult dogs were randomly divided into three groups. Eight dogs were assigned to the short training group, and they were allowed to run on a slope of 10? with the speed of 200m/min. Eight dogs were assigned to short intensified training group, and they were allowed to run on a slope of 10? with a speed of 300m/min. Four dogs served as controls, and they were allowed to have unrestricted activity in the cage. Dogs in training group and intensified group were asked to, run for an hour both in the morning and in the afternoon every day (tolally two hours per day). The training lasted for ten weeks. All the dogs were euthanized after ten weeks, and all the knee joints were harvested and examined histologically. Results Histological examination after ten weeks of exereises demonstrated that the signs of cartilage damage and repair were obvious in canine knee joint in training group and intensified group. Remodeling process was better promoted in the intensified group compared with training group, but there was no significant difference in Mankin score between these two groups. Conclusion Movements for a long time with great intensity are prone to induce sport injury. It is followed by repair process. Intensified training promotes a better remodeling reconstruction of articular cartilage than ordinary training.

Objective To explore the significance of such biological markers as cartilage oligomeric matrix protein (COMP), matrix metalloproteinases-1(MMP-1), matrix metalloproteinases-3 (MMP-3) and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) in serum and synovial fluid to evaluate early motional injury on knee joint cartilage induced by different modes of training. Methods Twenty adult dogs were randomly divided into three groups, i.e. common training group (n=8), short intensified training group (n=8) and control group (n=4). Magnetic resonance imaging (MRI) examinations were performed regularly (0, 2, 4, 6, 8, 10 weeks) to investigate the changes in the knee joint cartilage, while the concentrations of COMP, MMP-1, MMP-3 and TIMP-1 in serum and synovial fluid were measured by ELISA. Results It was revealed by MRI that the motional injury to knee joint cartilage occurred both in common training group and short intensified training group 2 weeks after training with different intensity, with severest injury occurring during 4~6 weeks after training, and then subsided gradually. The concentrations of COMP, MMP-1, MMP-3, TIMP-1 and MMP-3/TIMP-1 in serum and synovial fluid elevated in various degrees during training period with the lapse of time. Generally, the concentrations of MMP-1, MMP-3 and MMP-3/TIMP-1 in short intensified training group were lower, while that of COMP was higher, than that in common training group. The results suggested that the possibility of incurring articular injury in short intensified training group was lower than that in common training group, while the possibility of repairing the injury was on the contrary. Furthermore, the changes in levels of biomarkers in both serum and synovial fluid showed a close correlation. Conclusions Intensified training may result in moulding reconstruction better than the common training did. A combined determination of several biomarkers could be more sensitive than MRI examination on predicting the injury and repair of articular cartilage and on monitoring the course of illness. Therefore they are valuable for clinical application.

Allo-HSCT is a potentially curative therapy for many hematological malignancies,the basis of which is graft-versus-leukemia(GVL) effect.However,acute graft-versus-host disease(GVHD) is responsible for 15% to 40% of mortality and is the major cause of morbidity after transplantation.Therefore,separation of GVHD and GVL partially or completely could improve the transplant outcomes.This study focuses on the effects of G-CSF on T cells in peripheral blood stem cell grafts and bone marrow grafts and its mechanisms after in vivo G-CSF application.The separation of GVHD and GVL effect and the mechanisms of which after in vivo G-CSF and interleukin-11(IL-11) treatment of healthy donors were investigated.The main contributions of this research are listed as follows:(1) Immune tolerance of T cells was induced simultaneously in peripheral blood stem cell grafts and bone marrow grafts after in vivo G-CSF application;(2) T cell hyporesponsiveness and polarization of T cells from Th1 to Th2 were maintained after mixture of G-CSF-mobilized peripheral blood grafts(G-PB) and G-CSF-primed bone marrow grafts(G-BM) according to different proportions in vitro;(3) Treating donor with G-CSF and IL-11 decreased GVHD and retained GVL effect;(4) The incidence of acute GVHD was decreased after Allo-HSCT using G-PB and G-BM as allografts;(5) In combination with other techniques,the HLA barriers were overcomed using G-PB and G-BM as allografts;(6) The incidences of acute GVHD were significantly decreased and the GVL effects were retained or enhanced in relapsed patients after treatment by G-CSF-mobilized peripheral blood graft infusion compared with those received steady-state peripheral blood lymphocyte infusion,indicating that GVHD and GVL could be partially separated in clinical settings.Based on our results,we would conclude that the issues on the deficiency of donors are resolved,and novel strategies offered for the prophylaxis and treatment of patients with hematological malignancies who relapse after Allo-HSCT.Further studies on the mechanism of the separation of GVL and GVHD are warranted.

BACKGROUND:Traditional imaging examinations have lower sensitivity in diagnosing articular cartilage injury, and the effects of biomarkers in body fluid on diagnosing severity of cartilage injury should be evaluated.OBJECTIVE: To investigate the significance of the biomarkers, including cartilage oligomeric matrix protein (COMP), matrix metalloproteinases-1 (MMP-1), matrix metalloproteinases-3 (MMP-3) and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), in serum and synovial fluid in evaluating the early sports injury of articular cartilage.DESIGN: A randomized controlled trial.SETTING: the 105 Central Hospital of Chinese PLA.MATERIALS: Eighteen adult male hybrid dogs (15-25 kg, average 22.5 kg) were divided into three groups randomly: ①plain-running group (n =8): the dogs ran on the plain; ② slope-running group (n =8): the dogs ran on the slope of 10°; ③control group (n =2): the dogs could move freely.METHODS: Dogs in the plain-running group and slope-running group were trained daily, rurning for an hour in the morning and in the afternoon respectively(total two hours per day), 4 m/s, and the training lasted for 10 weeks. Serum and synovial fluid were collected on the dayof training, and at the 2nd, 4th, 8th and 10th weeks respectively in both the plain-running group and slope-running group, and the concentrations of COMP,MMP-1, MMP-3, TIMP-1 were detected with ELISA. Standard curves were built by the standard concentrations and A values, according to which corresponding concentrations of specimens were obtained. Knee joint MRI examination was performed, and the training was canceled on the day of examination. All the dogs were killed after 10 weeks, and the knee joint cartilages weight loading region of femur condyles were observed histologically.MAIN OUTCOME MEASURES: Concentrations of COMP, MMP-1, MMP-3, TIMP-1 in serum and synovial fluid on the day of training, and the 2, 4, 6, 8 and 10 weeks in each group; Results of MRI examination of knee joint; Results of the histological observation of knee joint cartilage after 10 weeks.RESULTS: All the 18 dogs were involved in the analysis of results. ① Results of examination in serum. ② Results of examination in synovial fluid: In the slope-running group, the COMP concentrations at 2, 4, 6, 8 and 10 weeks were obviously higher than those in the plain-running group (P ＜ 0.01), MMP-1 concentration at 2, 4 and 6 weeks were higher than those in the plain-running group (P ＜ 0.05-0.01), MMP-3 concentrations at 2, 4, 6 and 8 weeks were higher than those in the plain-running group (P ＜ 0.01);The MMP-3/TIMP-1 ratios at 2, 4 and 8 weeks were higher than those in the plain-running group(P ＜ 0.05-0.01).③ The concentrations of COMP,MMP-1,MMP-3 and TIMP-1 and the MMP-3/TIMP-1 ratio in serum were significantly correlated with the corresponding biomarkers in synovial fluid (r =0.631,0.502, 0.748, 0.651, 0.667, P＜ 0.01). ④ Results of histological observations: In both the plain-running group and slope-running group, obvious pathological changes of articular cartilage injury were observed at 10 weeks, the activity of articular chondrocytes in the surface layer was obviously decreased, which was severer in the latter group than in the former one. ⑤Results of MRI examination: Early sports injury changes of knee joint appeared at 2 weeks, and progressive aggravation occurred in both the plain-running group and slope-running group.CONCLUSION: Repetitive, impacted, torsional, shearing motions with long time and great intensity are easy to induce sports injury,even osteoarthritis.The sensitivity of these biomarkers is better than MRI in reflecting the pathological changes of articular cartilage.

As a high-throughout technique after DNA chips in biomedical research, protein arrays are developing, with high sensitivity and veracity. They have drawn increasing attention because they can provide a new technological platform for life science studies. There has been no effective diagnosis instrument for common articular cartilage injuries when they are at an initial stage, but when the disorders progress into an advanced stage, visible functional disturbances often arise. Protein arrays may offer a solution to the lasting tough problem in orthopaedic field. In this paper, we review the development and research of protein arrays, the regulation of proteins, their biochemical functions and their potential interaction partners, as well as changes of cartilage biomarkers in body fluid after injury. In addition, the feasibility and rationality of using protein microarrays for diagnosing early articular cartilage injuries are discussed.

Objective To understand common X-ray signs of bone and joint of forearm and shank,in order to provide a basis for diagnosis of skeletal fluorosis.Methods A total of 15 villages which were built for more than 50 years in Nongan and Qianan counties of Jilin province were selected and surveyed by quantitative epidemiological methods.These villages had no industrial and coal-burning-borne fluorine pollution; no other high fluoride sources; had not yet changed to low-fluoride drinking water,and had complete and stable historic data of fluoride content in drinking water.Drinking water fluoride content was determined.Inhabitants lived in the local for more than 10 years,between the age of 16-60 were chosen from the villages based on the 11 levels of fluoride content of drinking water from 0.5-7.0 mg/L and these people were divided into 5 age groups:16-,21-,31-,41-,51 to 60.In each age group,10-15 individuals were taken by stratified random sampling to do the forearm and shank joint X-ray examination.Drinking water fluoride content between 0.5-1.0 mg/L acted as the low fluoride group(control group),1.5-7.0 mg/L as the high fluoride group and various X-ray findings and the detection rate were observed.Results Three X-ray signs of skeletal fluorosis,joint degenerative change and osteoporosis were observed.Thereinto,joint degenerative change was the most common.In the high fluoride group(31.55％,153/485),the detection rate of joint degenerative change was higher than that of the low fluoride group (21.58％,41/190) and the difference was statistically significant (x2 =6.62,P ＜ 0.05).Seventy two of 675 peoples used by X-ray were detected with skeletal fluorosis,which occurred mainly in the higher intake of high fluoride group (x2 =25.65,P ＜ 0.01).When water fluoride was up to 6.0-7.0 mg/L,the detection rate of endemic skeletal fluorosis was increased to 44.92％(53/118).Regardless of higher or lower of water fluoride and mild or more serious of the state of fluorosis,changes of X-ray signs of bone surroundings happened first; on average with a detection rate of 95.83％ (69/72),the joint change followed by 79.17％ (57/72) and bone substance change the least at 56.94％(41/72),and the difference was statistically significant (x2=9.64,P ＜ 0.01).Regardless of water fluoride high and low,joint of elbow knee and wrist underwent degenerative change.Degenerative change of elbow joint happened first with a detection rate of 17.78％ (120/675),knee joint followed by 15.41％ (104/675),and wrist joint was the least at 7.85％ (53/675).The difference was statistically significant (x2 =30.74,P ＜ 0.01).Elbow degeneration detection rate(21.03％,102/485) in high fluoride group was significantly higher than that of the low fluoride group(9.47％,18/190; x2=12.47,P ＜ 0.01).X-ray examination showed 35 kinds of signs; most signs could be seen in the high fluoride and the low fluoride groups.But the changes of bone around were seen more often in higher fluoride content group than in lower fluoride content group.Conclusions X-ray signs of endemic skeletal fluorosis are complex and diverse.Only after comprehensive analysis,the diagnostic results are accurate and reliable.

Objective To explore the treatment to craniocerebral injury.Methods 6 patients suffered from serious, developing head injury with epidural and subdural hematoma were processed by cone socket hematoma drainage in ward before hematoma was eliminated.Results Hematoma drainage can be performed within 20 min after hospitalization. No case died and no infection occured. The score of Glasgow Outcome Scale were Ⅴ(2 cases), Ⅳ(2 cases),Ⅲ(1 case) and Ⅱ(1 case) 3 months after operation.Conclusion Cone socket hematoma drainage in ward is a good method to treat craniocerebral injury.

Objective:To study the effect of artificial gastric juice on the dissolution of schizandrin A to provide the parameters for the best extraction method of Schisandra chinensis. Methods:Schisandra chinensis was respectively extracted by artificial gastric juice and water. Schizandrin A in the extracts was determined by HPLC, and the dissolution of schizandrin A in artificial gastric juice and water was studied and compared. Results:At 60 min, schizandrin A dissolution was 0. 483% in artificial gastric juice, and 0. 362%in water. Conclusion:The dissolution of schizandrin A in artificial gastric juice is 33. 4% higher than that in water, suggesting artifi-cial gastric juice can significantly improve the dissolution of schisandrin A.

Objective To construct a recombinant lentiviral vector for human thymosin ?4 (TMSB4) and to test the mRNA and protein expression of TMSB4 in 293T cells after being infected by shRNA lentivirus. Methods We designed a specific sequence of small hair RNA targeting TMSB4 gene; the complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was cloned into the pGCSIL-GFP vector to construct a lentiviral vector. The vector was converted into the competent DH5a coli,which confirmed by PCR and sequencing. Then the viral vector and the packed systemic vector were cotransfected 293T cells to get the lentivirus. The virus titer was determined. Afterwards,in the 293T cells infected with the lentivirus,the expression of TMSB4 was detected by real time PCR and immunocytochemistry. Similarly,a primary fibroblast was also infected with the lentivirus. Results Compared with negative control cells,the mRNA and protein levels of TMSB4 in 293T cells infected with the lentivirus were reduced by 44% (0.56?0.11 vs 1.00?0.06,F=89.673,P

Objective To explore the immunologic composition and T cell function of rhG-CSF mobilized peripheral blood grafts(G-PB) and rhG-CSF primed bone marrow grafts(G-BM) after being mixed in different proportions.Methods T cell subgroups,dendritic cells(DC) and its subsets in G-PB and G-BM were analyzed by multicolor flow cytometry.Immunologic composition of G-PB and G-BM after mixture in different proportions were calculated,lymphocyte proliferation ability and quantities of interleukin-4(IL-4),interferon-?(IFN-?) secretion by T cells in mixture grafts were determined using MTT and sandwich ELISA assays.Results After co-incubation of G-PB with G-BM in different proportions,T cell proliferation in mixture grafts was lower than that of NG-BM and G-PB(P