OBJECTIVETo investigate development of a cell extraction process for preparing human meniscus acellular matrix, and morphology and biomechanical properties.

METHODSHuman meniscus were subjected to modified eight-step detergent, then, the specimens were assessed by staining with haematoxylin-eosin, toluidine blue, sirius red, saffron O, alcain blue and hoechst-33258, et al. The ultrastructure of the specimens was observed with scanning electron microscope. Transient recovery rate of deformation, maximal recovery rate of deformation and maximal compressive strength were tested to determine the biomechanical properties of the scaffold.

RESULTSEvery stain confirmed that the celluar constituents of the specimens were removed. The specimens stained positively by staining with sirius red. Lacuna were found irregularly not only on the surface of the meniscus,but also in the meniscus with scanning electron microscope. Pores in the specinmens were large, the diameter of pores was 80 to 760 microm, porosity was over 67%. The transient recovery rate of deformation was (89.62 +/- 1.04)%, the maximal recovery rate of deformation was 100% and the maximal compressive strength was (3.04 +/- 0.13)N, when the specimens were compressed 30%.

CONCLUSIONThe modified eight-step detergent can remove the immunogenic cell components from human meniscus, in addition, 3D extracellular matrix can be retained. The scaffold has good biomechanical properties. This scaffold stands a good chance to be an implant for future tissue engineering of the human meniscus.

Adult ; Cell Separation ; methods ; Cells ; chemistry ; cytology ; Cells, Cultured ; Humans ; Male ; Menisci, Tibial ; cytology ; Staining and Labeling

Urinary brain-derived neurotrophic factor (BDNF),an ubiquitous neurotrophin,was found to rise in patients with benign prostatic hyperplasia (BPH).We hypothesized that the urinary level of BDNF could be a potential biomarker for lower urinary tract symptoms (LUTS) in patients with BPH.Totally,76 patients with BPH-caused LUTS and 32 male control subjects without BPH were enrolled.International Prostate Symptom Score (IPSS) was applied to assess the symptom severity of LUTS.Urodynamic tests were performed for the diagnosis of underlying detrusor overactivity (DO) in the patients with BPH.Urine samples were collected from all subjects.Urinary BDNF levels were measured using enzyme-linked immunosorbent assays and normalized by urinary creatinine (Cr) levels.Seventy-six BPH patients were divided into moderate LUTS group (n=51,7＜IPSS ≤ 20) and severe LUTS group (n=25,IPSS＞20) according to the IPSS.Of the 76 BPH patients,DO was present in 34 (44.7％)according to the urodynamic test.The urinary BDNF/Cr levels were significantly higher in BPH patients with moderate LUTS (8.29±3.635,P＜0.0001) and severe LUTS (11.8±6.44,P＜0.0001) than normal controls (1.71±0.555).Patients with severe LUTS tended to have higher urinary BDNF/Cr levels than patients with moderate LUTS (11.8±6.44 vs.8.29±3.635,P=0.000).The conditions of BPH with LUTS correlated with elevated urinary BDNF levels,and urinary BDNF levels were even higher in BPH-DO patients.The results of this study have provided evidence to suggest that urinary BDNF level test could evaluate the severity of LUTS in BPH patients,and BDNF level can be used as a biornarker for the diagnosis of DO in BPH patients.

BACKGROUND: Deep vein thrombosis (DVT) of the lower limbs is one of the severe complications of total hip replacement (THR)during the perioperative period. The incidence rate was high. The effect of primary disease on the DVT after THR in the elderly remains poorly understood.OBJECTIVE: To explore the effect of primary disease on DVT after THR in the elderly.METHODS: 147 cases with unilateral THR aged 64-93 years, were included and divided into two groups. Fracture group contained 68 cases, which of them were all traumatic femoral neck fracture ones. Osteopathia group contained 79 cases, and they had no traumatic injury. We selected total hip prostheses according to their physiological age, preoperative socialization ability,substantia ossea and life expectancy. Biological prosthesis in 5 cases and mixed prosthesis in 12 cases were used, while the others used bone cement prosthesis. If the patient had pain and/or swelling on the injured limb, with or without Homans/Neuhofs sign, we did the pressurized ultrasonic Doppler to examine whether the patient had got DVT.RESULTS AND CONCLUSION: In the fracture group, 32 cases had swelling in the injured limbs, and 20 cases with pain, 15 cases with Homans/Neuhofs sign, and 29 DVT cases were confirmed by the pressurized ultrasonic Doppler. One femoral neck fracture case of THR had no DVT clinical signs and was dead 17 days later. The autopsy found that it was an mixed type combined of pulmonary embolism; Osteopathia group: 20 cases had swelling in the injured limbs, and 11 cases of pain, 9 cases of Homans/Neuhofs sign, 20 DVT cases were confirmed by the pressurized ultrasonic Doppler. The blood coagulation state was greater, and the incidence rate of DVT in the lower limbs was greater in the fracture group compared with osteopathia group (P ＜ 0.05). These indicated that femoral neck fracture is a high risk factor for DVT development in lower limb affer THR in the elderly.

Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Nucleus ; genetics ; Cells, Cultured ; Diploidy ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; metabolism ; ultrastructure ; Transfection

Objective:To evaluate the fit of titanium alloy removable partial denture framework fabricated by selective laser melting(SLM) technique.Methods:7 Kennedy Ⅲ Ti-6Al-4V removable titanium alloy partial denture frameworks were fabricated by SLM technology.An optical scanner was used to scan the gypsum model adhered with the silicone rubber film obtained by the impression method before and after removal of the film.Geomagic Qualify 2013 software was used to analyze the gap between the model and the tissue surface of the major connector of the framework,the fit of the frame work was evaluated.Results:The overall 3D deviation between the titanium alloy frameworks and the gypsum model was (0.221 9±0.07) mm.Conclusion:The fit of the titanium alloy removable partial denture framework made by SLM technology can basically met the clinical requirements.

Purpose:To compare the therapeutic and adverse effects between continuous and intermittent infusion of fluorouracil(5-FU) in advanced colorectal cancer.Methods:17 patients of advanced colorectal cancer were treated with continuous intravenous infusion of 5-FU 2.5g in 100 ml of 5% glucose in a continuous infusions pump,for 120 h (Group A).16 patients of similar severity were treated with 5-FU 500 mg in 500 ml of 5% glucose 2 h iv daily for 5 d (Group B). Total of 33 patients were treated with calcium folinate (CF) 200 mg in 100 ml of 5% glucose 1 h iv daily for 5 d and HCPT 10 mg in 250 ml of 5% glucose 2 h iv daily for 6d.The therapeutic and adverse effects were evaluated according to the cri- teria recommended by WHO.Results:Effectiveness was noted in 9 patients in Group A and 3 patients in group B.(x~2 test P

Objective To investigate the clinical manifestations and treatment regiment of children with juvenile dermatomyositis(JDM).Methods The clinical manifestation,changes of serum muscale enzyme,myopathic laboratory examination,treatment and prognosis of 15 children with JDM retrospectively admitted from Jan.1990 to Jan.2004 were analyzed.Results All of the children had symmetrical weakness of the proximal muscles.The most frequent features were heliotrope and Gottron's papules.Elevated muscle enzymes were noted in all cases.Electromyography revealed typical change of myopathic type and muscle biopsy was compalible with myositis in all cases.Most of patients achieved normal muscle enzymes within 1 month and had improved muscle strength with 2.5 monthes of the initiation of corticosteroid therapy.Conclusion It is very important to know the clinical features of JDM,and prompt diagnosis and treatment will result in an improved prognosis.

AIM: To seek a small interfering RNA ( siRNA ) sequence targeting rat inhibitor of nuclear factor kappa Bα ( IκBα) that can specifically and effectively suppress IκBα mRNA expression of rat ciliary muscles in vivo.METHODS:Three IκBα specific double stranded siRNAs were designed and synthesized. They were transfected into rat A7r5 cells which express IκBα gene. Flow cytometry was used to assess transfected efficiency. The mRNA and protein levels of IκBα were examined by Real Time quantitative polymerase chain reaction ( Real Time-PCR ) and western blot to screen a candidate valid sequence with the highest inhibitory rate. The Cy3 labeled non-specific control siRNA or the candidate valid siRNA was then injected into rat anterior chamber. Distribution of Cy3- siRNA in rat ciliary muscles was viewed by fluorescence microscopy, and the inhibitory effect in vivo of the valid siRNA was identified via Real Time-PCR and immunofluorescence. RESULTS: The suppression effect of the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene was most obvious by vitro screening. By anterior chamber injection, this valid siRNA could reach rat ciliary muscles and effectively suppress IκBα gene expression with the highest inhibitory rate of 59. 0% on mRNA level at 24h after RNAi, and 52. 3% on protein level at 72h after RNAi (P<0. 01).CONCLUSION: It proves that the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene is the valid sequence to suppress rat IκBα expression of ciliary muscles by RNAi in vivo.

Objective To compare the effects of short- term high- intensity interval training (HIT) and moderate continuous training (MCT) on inflammatory mediators for healthy young males. Methods 19 healthy young male volunteers were randomized into HIT group (n=6), MCT group (n=7) and no training control (CON, n=6) group. The groups of HIT and MCT accepted treadmill exercise once per 2 days to 6 times (2 weeks), and the CON group did not accepted treadmill exercise. The levels of high-sensitive C-reactive protein (hs-CRP), interleukin (IL) -1α, IL-6 and tumor necrosis factor α (TNF-α) were detected 3 days before exercise, immediately after the first exercise, 3 days after the course. Results There was no difference among groups in the level of hs-CRP, IL-1α, IL-6 and TNF-α in all the time (P>0.05). The concentration of IL-1α increased in the MCT group after the first exercise (P<0.01), and also the concentration of IL-6 in the CON group after the course (P<0.05). Conclusion It needs further srudy for the significance of HIT and MCT in the levels of inflammatory media.

OBJECTIVE: To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle mechanical injury in rats. METHODS: The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury) and control group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. RESULTS: At post-injury 6-12h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN decreased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14d, the percentage of FBC reached peak. CONCLUSION The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.
Animals ; Fibroblasts ; Muscle, Skeletal/injuries* ; Neutrophils ; Rats ; Time Factors