OBJECTIVETo study effects of autogenous femoral head bone grafting combined with total hip replacement for the treatment of old center dislocation of hip joint, and to evaluate the superiority of acetabular defect reconstruction with autogenous bone.

METHODSFrom March 2008 to December 2013, 16 patients (16 hips) with old center dislocation of hip joint were treated with autologous bone graft combined with total hip replacement. There were 11 males and 5 females, ranging in age from 41 to 72 years old, with an average of 56.3 years old. The duration of the disease ranged from 3.6 to 37.2 years. Twelve patients had injuries caused by traffic accidents and 4 patients were caused by falling down. The hip joint pain at the first month after operation, Harris score and hip joint movement of all patients were observed by using electronic case follow-up system.

RESULTSAll the patients were followed up, and the duration ranged from 11 to 78 months, with an average of 27.3 months. The postoperative hip joint movement of (56.2±23.4)° VAS 86.3±7.2 and Harris score 32.6±12.6 were all better than preoperative (181.8±17.6)°, 11.1±2.6 and 86.3±7.2. The joint pain was reduced and the function limitation was improved after operation, and the prosthesis position was good.

CONCLUSIONWhen the autologous femoral head bone grafting combined with total hip arthroplasty is used to treat old hip central dislocation, the initial and long-term stability of acetabular bone grafting is good, and the technique can avoid allogeneic bone complications, bone source rationally and reduce the economic burden of patients.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femur Head ; transplantation ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Transplantation, Autologous

Objective To investigate the changes in cortical excitability and inhibitory circuits of patients with Alzheimer's disease (AD) or behavioral variant frontotemporal dementia (bvFTD) using transcranial magnetic stimulation (TMS).Methods Forty-four patients with AD,30 patients with bvFTD and 44 healthy controls were enrolled in the study.The epidemiological data of all subjects were collected.Each participant received a neurological evaluation and neuropsychologic tests which included Mini-Mental State Examination (MMSE),Activities of Daily Living (ADL) and Hamilton Rating Scale for Depression (HAMD).Neurophysiological evaluations including resting motor threshold (rMT),active motor threshold (aMT) and cortical silent period (CSP) were conducted by means of TMS.Neurophysiological parameters were compared among groups.Results There were significant differences in MMSE,ADL and HAMD assessments among 3 groups,but no significant differences between AD and bvFTD groups.There were significant changes in left rMT(46.52％ ± 8.77％,52.00％ ± 7.30％ and 52.14％ ± 8.27％,F =6.295,P=0.003),left aMT(29.68％ ±7.01％,35.13％ ±8.46％ and 35.39％ ±7.24％,F=7.735,P=0.001) and right rMT(47.82％ ±7.94％,52.07％ ±8.77％ and 53.12％ ±8.61％,F=4.772,P=0.010) among 3 groups.AD patients showed significantly reduced left rMT and aMT comparing to bvFTD patients (P =0.017 and 0.008 respectively) and controls (P =0.005 and 0.002 respectively).In addition,AD patients had a significant decrease in right rMT comparing to controls (P =0.011).There were no statistically significant differences in TMS parameters between bvFTD patients and controls.Conclusions AD is associated with hyperexcitability of the motor cortex,whereas the lack of changes in motor cortical excitability is found in bvFTD.TMS technique may be helpful in differential diagnosis between AD and bvFTD.

Objective To investigate the clinical characteristics of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA) associated vasculitis (MPO-AAV) with pulmonary injury and the relationship between pulmonary injury and ANCA against light chain of MPO (LCMPO-ANCA).Methods A total of 195 patients with newly diagnosed primary active MPO-AAV were recruited in this prospective study.Indirect immunofiuorescence assay was used to detect peri-nuclear ANCA (p-ANCA).Immunoblotting and ELISA were used to detect myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA).Clinical features of patients with both positive p-ANCA and MPO-ANCA were collected.Disease activity was evaluated by Birmingham Vasculitis Activity Score-version 3 (BVAS-V3) Recombinant light chain of MPO was used to coat substrate of LCMPO-ANCA by ELISA.The clinical characteristics of pulmonary injury and its correlation with serum levels of p-ANCA,MPO-ANCA and LCMPO-ANCA were explored.Results All 195 patients (64 male and 131 female),consisted of 191 patients (98.0％) with microscopic polyangiitis,3 patients (1.5％) with granulomatosis with polyangiitis,and 1 (0.5％) with eosinophilic granulomatosis with polyangiitis including 64 men and 131 women.Their mean age was (63.2 ±13.5) years old.The level of MPO-ANCA had a positive correlation with general BVAS-V3 (r =0.193,P =0.007) in all patients,and the level of LCMPO-ANCA was positively related with the pulmonary BVAS-V3 (r =0.228,P =0.001).As for multiple systemic damages,the incidence of lung involvement was 60.51％(118/195),which ranked second to renal involvement (71.80％,140/195).The most common pulmonary injuries represented as pulmonary infiltration of 80.51％ (95/118),pleural effusion / pleurisy of 41.53％(49/118),pulmonary nodule or cavity of 22.03％ (26/118).Compared with those without lung involvement,the patients with pulmonary injuries were older [(66.39 ± 10.70) years old vs (58.30 ±15.72) years old;t =4.277,P =0.001],had a shorter course of disease [2.00(1.00,10.50) months vs 3.00(1.00,3.50) months;t =-2.283,P=0.024],and higher scores of general BVAS-V3 (18.21 ±6.08 vs 15.18 ± 5.64;t =3.501,P =0.001).Also,in the patients with pulmonary lesions,the positive rate of LCMPO-ANCA was significantly higher (35.59％ vs 6.49％;x2 =21.569,P ＜ 0.001),and the level of LCMPO-ANCA was significantly higher (0.377 ±0.229 vs 0.285 ±0.079;t =3.399,P =0.001)than those without lung involvement.The pulmonary BVAS-V3 in the patients with LCMPO-ANCA was significantly higher than that in the patients without LCMPO-ANCA (4.34 ± 2.10 vs 2.59 ± 2.52;t =4.301,P ＜ 0.001),whereas the pulmonary BVAS-V3 was not correlated with LCMPO-ANCA (r =0.035,P =0.708) in patients with lung injuries.Conclusion Pulmonary injury was relatively common and insidious in patients with MPO-AAV.To monitor ANCA level is necessary in patients with pulmonary injury.LCMPO-ANCA might play an important role in the pathogenesis of pulmonary lesions in AAV.

Objective To assess the feasibility of chondrocyte and osteoblast composites in vitro cultured in bioreactor in repairing cartilage defects.Methods Marrow mesenchymal stem cells were isolated and cultured in vitro,and then were induced to chondrocytes and osteoblasts by growth factor.Chondrocytes and osteoblasts were cocultured in bioreactor for 21 days to form the composites.The adhesion,extension and proliferation of chondrocytes and osteoblasts were observed under scanning electron microscope.The cartilage defects on canine model were repaired with the chondrocyte and osteoblast composites.Results The induced chondrocytes and osteoblasts had fine adhesion,extension and proliferation in the β-TCP scaffold.The repaired tissues in experimental group maintained their thickness to the full depth of the original tissues.A statistical difference was observed between negative control group and experimental group(q=12.337 0,P ＜ 0.01)and between blank control group and experimental group (q=31.539 3,P ＜0.01).Conclusion Perfusion bioreactor makes chondrocyte and osteoblast survive and proliferate in a three-dimensional scaffold and increases the composition rate of the chondrocyte and osteoblast.

BACKGROUND: In recent years, ultrasound microbubble gene transfer system has been applied for gene transfection in many parts of the body, but it has been seldom reported to be used for gene transfection in bone parts. OBJECTIVE: To investigate the efficiency and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits.METHODS: Japanese big-ear rabbits were randomly divided into five groups: bare transfection, pre-irradiation + bare transfection, ultrasound transfection, pre-irradiation+ultrasound transfection, and repeatable transfection. In the first two groups, ultrasound-targeted gene transfection and irradiation was not used, but in the latter three groups, ultrasound-targeted microbubble destruction was used to transfect enhanced green fluorescent protein (EGFP) plasmid into the femoral head of rabbits. At 1 week after transfection, EGFP expression in femoral head was observed under the fluorescence microscope. RESULTS AND CONCLUSION: EGFP expression appeared in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection. The transfection efficiency of EGFP plasmid was significantly higher in the repeatable transfection group than in the other groups (P < 0.01). Obvious injury loci were not observed in the soft tissue and bone tissue slices of ultrasonic irradiation parts in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection groups. These results confirm that ultrasound-targeted microbubble destruction is a safe and effective method to transfect EGFP plasmid into the femoral head of rabbits.

Objcctive To detect the telomerase activity in rat hepatic oval cells, and to explore the relationship between telomerase expression and the proliferation and differentiation of oval cells. Methods The 2-acetamidofluorene/partial bepatectomy (2-AAF/PH) rat model was used to induce the proliferation of oval cells. Oval cells were isolated by modified collagenase perfusion and gradient centrifugation. Electron microscope exami-nation and immunofluorescence were adopted to identify oval cells. Immunohistochemistry, RT-PCR and fluores-cence quantitative PCR were used to detect the expression of telomerase in oval cells. All the data collected were analyzed by t test. Results The proliferatiun of oval cells was successfully induced by 2-AAF/PH rat model. Freshly isolated oval cells showed a large and ovoid nuclei, a small proportion of cytomplasm and a cobblestone appearance. When viewed by electron microscopy, there were few and immature organelles, and the nucleus/ cytoplasm ratio was high. Immunofluorescence staining showed that oval cells expressed OV-6, alpha-fetoprotein, cytokeratin-19, albumin and c-kit. Telomerase reverse transcriptase (TERT) was located in the nuclei of oval cells which were around the portal areas. As oval cells differentiated into small hepatocytes, the number of TERT-positive cells decreased significantly. The expression level of TERT mRNA in normal rat liver tissue was 2.27 times higher than that in LE-6 oval cells; the expression level of TERT mRNA in the isolated oval cells was 1.26 times higher than that in LE-6 oval cells. The telomerase activity decreased gradually (from △A=1.05, 1.15 to △A=0.25, 0.45) as the increase of oval cells passage (from passage 24 to passage 40) (t=17.74, 12.38, P<0.05). Conclusions Oval cells have telomerase activity. Telomerase may be indispensable for maintaining the proliferative and multi-directional differentiation abilities of oval cells.

OBJECTIVEWith functional magnetic resonance imaging (fMRI), cerebral function imaging of acupuncture and sham acupuncture at Taichong (LR 3) in patients with essential hypertension (EH) was observed to analyze central mechanism of Taichong (LR 3) to treat EH.

METHODSEighteen EH patients were divided into a twisting acupuncture group (10 cases) and a sham acupuncture group (8 cases), which were treated with twisting manipulation and sham acu- puncture at right Taichong (LR 3), respectively. The treatments were both given with 30-second manipulation and 30- second interval for totally 5 min. During the acupuncture, fMRI was adopted to scan the cerebral function imaging of EH patients on resting state, and the acquired data were analyzed with SPM2 module in Matlab software.

RESULTSAcupunc- ture at Taichong (LR 3) in EH patients mainly increased opposite-side oxygen consumption, which generally activated left anterior cingulated gyrus (BA 32), left inferior parietal lobule (BA 40), left inferior temporal gyrus (BA 19), left middle temporal gyrus (BA 37) and right anterior central gyrus (BA 6). The activated areas were related to motor, vision and cognition of emotion. Stimulation region of sham acupuncture mainly included right anterior cingulated gyrus (BA 24), left inferior parietal lobule (BA 40) and left inferior frontal gyrus (BA 7).

CONCLUSIONThe central mechanism of Tai- chong (LR 3) on antihypertensive effect and symptom improvement is not in thalamus and medulla that are traditionally believed to control blood pressure. Taichong (LR 3) could activate anterior cingulated gyrus to regulate parasympathetic nerve and lower blood pressure, and through anterior cingulated gyrus, the connection with the surrounding areas is strengthened to improve the cognitive impairment caused by long-term hypertension.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Brain ; diagnostic imaging ; Cognition ; Essential Hypertension ; Female ; Humans ; Hypertension ; diagnostic imaging ; psychology ; therapy ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography

Objective To construct p66shc gene interfering lentivirus vectors recombination and transfect it to 293T cells ,RNA interfering was carried out to induce p66shc gene silence ,so as to provide basis for further study of the p66shc function .Methods Screening of three RNA targets which were named after p66shc‐shc1 ,p66shc‐shc2 ,p66shc‐shc3 ,cloned into the pLenR‐GPH vec‐tor ,which contained green fluorescent protein(GFP) and transformed into DH5αcells .The positive clone were picked out for right sequencing and transfected to 293T cells with pRsv‐REV ,pMDlg‐pRRE ,pMD2G .The expression of GFP in inverted fluorescence microscope confirmed the virus packaging success .Fluorescence quantitative PCR and Western blot technology were used to investi‐gate the expression of p66shc at the molecular and protein levels ,p66shc‐shc1 target of effective silencing p66shc gene was selected to prepare for subsequent tests .Results The shRNA lentivirus vector was constructed which could express p66shc and was trans‐fected into 293T cells successfully .Fluorescence quantitative PCR and Western blot technology were used to investigate p66shc gene silence by RNA interference .Conclusion The lentivirus RNAi vector of targeted expression p66shc could induce p66shc gene si‐lence at the molecular and protein levels after transfected into 293T cells by RNA interference .

Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.

Objective To investigate the expression of CD55 and myeloperoxidase (MPO) on neutrophils in patients with MPO-specific anti-neutrophil cytoplasmic antibody associated vasculitis(MPO-AAV), and analyze the relationship between the expression and clinical manifestation.Methods Forty untreated patients with active MPO-AAV (patient group) and 30 healthy volunteers (control group) were enrolled in this study.The CD55 on neutrophils and both membrane and cytoplasmic MPO were detected by flow cytometry.Serum fragment-from the activated complement factor B(Ba) and MPO were measured by ELISA.The clinical activity of vasculitis was valued by Birmingham vasculitis activity score-version 3(BVAS-V3).The significance of laboratory data was evaluated by Spearman correlation test and multivariate linear regression analysis.Results (1)The mean fluorescence intensity(MFI) of CD55 expressed on neutrophils was significantly higher than that in control group[4 068.6±2 306.0 vs 2 999.5±1 504.9,P=0.033].Similar results of serum MPO and Ba in patient group were found compared to controls [500.0(381.0, 612.7) IU/L vs 286.9(225.5, 329.1) IU/L,P<0.001;35.2(25.2, 79.5) ng/L vs 18.0(15.0, 28.0) ng/L,P<0.001], respectively.However, MIF of cytoplasmic MPO in patients was significantly lower than that of control group(1 577.1±1 175.9 vs 3 105.3±2 323.0,P=0.003).(2) In patient group, cytoplasmic intensity of MPO was negatively associated with the serum levels of MPO(r=-0.710,P<0.001) and Ba (r=-0.589,P=0.001).Moreover, serum MPO was positively associated with serum Ba(r=0.691,P<0.001).Membrane intensity of CD55 on neutrophils was positively correlated with patient age (r=0.514, P=0.001), C reactive protein (r=0.376, P=0.018), peripheral neutrophils count (r=0.485, P=0.001) and BVAS-V3 (r=0.484, P=0.002), whereas negative correlation between membrane CD55 and disease duration was seen (r=-0.403,P=0.01).(3) The result of multiple linear regression analysis showed there was statistically significant positive correlation between MFI of CD55 expressed on neutrophils and BVAS-V3 (β=0.001,P=0.027).Conclusions In MPO-AAV,CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking AAV, CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking by complement alternative pathway.Activated neutrophils release more MPO and lysosome to intensify the inflammation reaction and aggravate the disease.Thus CD55 might become a new potential target for the treatment of this disease in the future.