Objective To explore new insight into the immunopathogenesis of ulcerative colitis.Methods Immunohistochemistry for the immunoglobulin(IgG)and complement(C 3c )was performed in biopsy specimens of the mucous membrane from 50 patients with ulcerative colitis(32 in active state and 18 in inactive state)and 5 controls.Results The immunoglobulin and activated complement were found in the epithelium mucosae and basement membrane of capillary walls in ulcerative colitis,with highest density in patients with active ulcerative colitis as compared with inactive ones.Conclusions The activation of immunoglobulin(IgG)and complement(C 3c )are closely related to active ulcerative colitis which plays an important role in the tissue damage of active ulcerative colits.Immunoglobulin(IgG)and complement(C 3c )are important components of immunoregulatory network of ulcerative colitis

Objective To investegate the effect of ginsenoside Rg1 on the apoptosis related protein FLICE-inhibitory protein(FLIP),Fas-associated death domain protein (FADD)and Caspase-3 in the subatania nigra(SN)of 1-methyl-4-phenyl-1,2,3,6-tetrahyd-ropyridine (MPTP)-induced mouse models of Parkinson’s disease(PD), and to investigate the role of FADD and FLIP in the pathogenesis of PD and the protective effect of ginsenosides Rg1 on dopaminergic neurons.Methods 45 C57BL/6N mice were randomly divided into control group,model group and ginsenoside Rg1 group (n=15).The mice in model group were injected with MPTP by intraperitoneal,the mice in Rg1 group were injected with ginsenoside Rg1 before injecting MPTP,and the mice in control group were injected with normal saline by intraperitoneal. The behavioral changes of the mice in various groups were observed, and immunohistochemistry and Western blotting methods were used to observe the expressions of tyrosine hydroxylase (TH),FADD,FLIP and Caspase-3 in substantia nigra of the mice.Results Compared with control group,the mice in model group presented with typical symptoms of PD, the TH-positive neurons in the subatania nigra was significantly reduced (P<0.01 ), the number of FADD, FLIP and Caspase-3 positive cells was significantly increased(P<0.01),and the cytoplasm was deeply stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly increased (P<0.01).Compared with model group,the PD symptoms of the mice in ginsenoside Rg1 group reduced, the number of TH-positive neurons was significantly increased, the number of positive cells of FLIP,FADD and Caspase-3 were significantly reduced(P<0.01),and the cytoplasm was lightly stained;the protein expression levels of FADD, FLIP and Caspase-3 were significantly reduced (P<0.01 ). Nonlinear correlation analysis found that there was a positive relationship between the number of FADD and Caspase-3 positive cells (r=0.791,P<0.05).Conclusion Ginsenoside Rg1 may play a neural protective effect dopaminergic on neurons by modulating the FADD and FLIP expressions in SN of PD model mice.

Objective:To study the influence of bone morphogenetic protein-7 ( BMP-7 ) on chondro-cyte secretion and expression of type Ⅱ collagen ( Col-Ⅱ) , aggrecan ( AGG ) and SRY-related high mobility group-box gene 9 ( Sox9 ) mRNA in porous tantalum-chondrocyte composites.Methods: The articular chondrocytes were isolated from 3-week-old New Zealand immature rabbits and identified.The 2nd generation of chondrocytes with 1 ×106/mL inoculate concentration was seeded in porous tantalum and divided into 4 groups, and control group ( tantalum/chondrocyte) , 50μg/L BMP-7 group (50μg/L BMP-7/tantalum/chondrocyte) , 100 μg/L BMP-7 group ( 100 μg/L BMP-7/tantalum/chondrocyte ) , and 200 μg/L BMP-7 group ( 200 μg/L BMP-7/tantalum/chondrocyte ) .The proliferation of chondro-cytes was measured by CCK-8 assay.The chondrocyte growth and morphology were observed by scanning electron microscopy ( SEM) .The synthesis of glycosaminoglycan ( GAG) in chondrocytes was tested by dimethyl methylene blue ( DMMB) colorimetric quantification method.Col-Ⅱ, AGG and Sox9 mRNA in chondrocytes were detected by real-time PCR.Results: The chondrocytes were spindle-shaped in 24 hours of primary cell culture and most cells became polygonal shaped in 4 days.The chondrocytes were affirmed by alcian blue, safranin O and Col-Ⅱimmunocytochemistry staining.The result of CCK-8 assay showed that the level of cell proliferation in 100 μg/L BMP-7 groups were higher than those in the other groups ( P<0 .05 ) .The chondrocytes implanted into porous tantalum scaffolds with BMP-7 had better functions, by which cytoplasmic processes developed and extended to the surface and inner of porous tan-talum by SEM observation.DMMB quantitative determination of GAG showed that GAG amount of chon-drocytes in 100 μg/L BMP-7 groups was significantly higher than those in the other groups ( P<0 .05 ) . The expressions of Col-Ⅱ, AGG and Sox9 mRNA in chondrocytes were up-regulated in the experimental groups, compared with the control group and the best effect appeared when concentration of BMP-7 was 200μg/L.(P<0.05).Conclusion:BMP-7/tantalum/chondrocytes composites enhanced in vitro chon-drocyte proliferation and extracellular matrix greatly, and can promote chondrogenic gene expression.

BACKGROUND:Bone morphogenetic protein 7 (BMP-7) can induce bone and cartilage formationin vivo, and induce chondrogenic and osteogenic differentiation of mesenchymal cels in muscles and around the vessels.
OBJECTIVE:To observe the structure of domestic tantalum-muscle interface fibrous capsule, growth of muscle and smal blood vessels into the porous tantalum and the ability of ectopic osteogenesis after implantation of porous tantalum loaded with BMP-7 into the erector spinae of rabbits.
METHODS: Porous tantalum slices loaded with BMP-7 (experimental group) and porous tantalum slices (control group) were implanted into the erector spinae muscle of New Zealand white rabbits. And the porous tantalum slices with surrounding muscle tissues about 0.5 cm thick were removed at 2, 4, 8 weeks after implantation, and observed under scanning electron microscope for hematoxylin eosin staining, Masson staining and hard tissue slice observation.
RESULTS AND CONCLUSION:(1) Hematoxylin-eosin staining: Fibrous capsule formation was observed around the materials in the two groups, and with the extension of time, the fibrous capsules were slightly dense, and thinned. There was no obvious inflammatory reaction in the interface between the material and the muscle. There was no significant difference between the two groups in the fibrous capsules thickness. (2) Scanning electron microscope: 2 weeks after the surgery, a smal amount of colagen and muscle fibers were formed in the porous tantalum pores in the two groups, and some of colagen fibers attached to the pore wals. At 8 weeks after the surgery, al the pores of porous tantalum were ful of muscle fibers that were combined with the pore wal closely. There was no significant difference between the two groups. (3) Hard tissue slices: 2 weeks after the surgery, a smal amount of fibroblast cels and muscle fibers grew into the pores of porous tantalum in the two groups and new capilaries grew into the pores of porous tantalum in the experimental group. At 8 weeks after the surgery, the porous tantalum and al the pores were ful of muscle fibers that were combined with the pore wal closely, the number of smal blood vessels and cels decreased, and the tantalum and the muscle were fused closely. (4) Masson staining: 8 weeks after the surgery, a large number of mesenchymal cels, ossein and cartilage matrix formed in the muscle gaps and a few cartilage bone tissues were formed in the experimental group, but no cartilage was found in the control group. The study showed that porous tantalum carrying BMP-7 has good biocompatibility and osteogenic induction ability. Subject headings: Tantalum; Bone Morphogenetic Protein 7; Tissue Engineering.

BACKGROUND: At present, there is evidence that domestic porous tantalum has good biocompatibility and osteogenic properties, but the specific osteogenic mechanism and its effect on osteogenic factors are still unclear. OBJECTIVE: To observe the effects of domestic porous tantalum materials on the expression of collagen type I, tissue transglutaminase-2 and calcium-binding protein A4 in MG63 cells. METHODS: MG63 cells in logarithmic growth phase were inoculated onto 24-well plates and cultured in three groups: in blank group, conventional medium was added; in tantalum extract group, porous tantalum material extract was added; and in tantalum scaffold group, porous tantalum material and conventional medium were added. On 1, 3, 5, 7 and 9 days of culture, the cell proliferation of each group was detected by cell counting kit-8 method. On 5 days of culture, the levels of collagen type I, tissue transglutaminase-2 and calcium-binding protein A4 secreted by MG63 cells in each group were detected by ELISA. Western blot assay was used to detect the expression of three proteins in each group. RESULTS AND CONCLUSION: (1) With the prolongation of culture time, the number of cells in each group increased gradually. There was no difference in cell proliferation among the three groups at different time points (P> 0.05). (2) The secretory levels of collagen type I and tissue transglutaminase-2 in the tantalum scaffold group were significantly higher than those in the blank group and tantalum extract group (P < 0.05), while the secretion of collagen type I and tissue transglutaminase-2 in the tantalum extract group was significantly higher than that in the blank group (P < 0.05). The secretion of calcium-binding protein A4 in the tantalum scaffold group was significantly lower than that in the other two groups (P < 0.05). (3) The expression of collagen type I and tissue transglutaminase-2 protein in the tantalum scaffold group was significantly higher than that in the blank group and tantalum extract group (P < 0.05), while the expression of collagen type I and tissue transglutaminase-2 protein in the tantalum extract group was significantly higher than that in the blank group (P < 0.05). The expression of calcium-binding protein A4 in the tantalum scaffold group was significantly lower than that in the blank group and tantalum extract group (P < 0.05). To conclude, domestic porous tantalum materials could promote the secretion of collagen type I and tissue transglutaminase-2 by MG63 cells, and inhibit the secretion of calcium-binding protein A4.

PURPOSE: In this study, we aimed to evaluate lymphocyte-activation gene-3 (LAG-3) expression and its prognostic value in neoadjuvant-treated triple-negative breast cancer (TNBC). METHODS: LAG-3, programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), and CD8⁺ tumor-infiltrating lymphocyte (TILs) levels were examined using immunohistochemistry in 148 preand 114 post-neoadjuvant chemotherapy (NACT) specimens of human TNBC tissue. Correlations between expression levels and clinicopathological features were analyzed. Prognostic values for combined detection in TNBC following NACT were evaluated. RESULTS: In pre-NACT specimens, LAG-3 expression showed a significant association with pathological complete response (pCR, p=0.038) and was correlated with PD-1 (p<0.001) and PD-L1 (p=0.008). In post-NACT specimens, high expression of LAG-3 showed significant effects on nodal status (p=0.023) and PD-1 (p<0.001). The expression of immune markers on TILs significantly increased following NACT. Multivariate analysis indicated that only nodal status (odds ratio [OR], 0.226; 95% confidence interval [CI], 0.079–0.644; p=0.005) and high quantities of CD8⁺TILs (OR, 3.186; 95% CI, 1.314–7.721; p=0.010) are independent predictors of pCR. Nodal status (hazard ratio [HR], 2.666; 95% CI, 1.271–5.594; p=0.010), CD8⁺TILs (HR, 0.313; 95% CI, 0.139–0.705; p=0.005), and the LAG-3-high/PD-L1-high group (HR, 2.829; 95% CI, 1.050–7.623; p=0.040) provided prognostic values for patients with TNBC following NACT. CONCLUSION: CD8+TILs were sensitive predictive markers in response to NACT. High expression of LAG-3 in residual tissues, especially in combination with PD-L1, was associated with poor prognosis.
Biomarkers ; Drug Therapy ; Humans ; Immunohistochemistry ; Lymphocytes, Tumor-Infiltrating ; Multivariate Analysis ; Neoadjuvant Therapy ; Polymerase Chain Reaction ; Prognosis ; Triple Negative Breast Neoplasms*

Clinical treatment of large segmental bone defect has always been a major challenge for the medical community,which is due to the complex and diverse pathogenic factors of large segmental bone defect.In recent years,the clinical treatment of large segmental bone defect has made great progress,and the main treatment options include vascularized or non-vascularized autologous bone grafts,allograft bone transplantation,masquelet technology,llizarov technology and bone tissue engineering.Therefore,understanding the advantages and disadvantages of various treatment options is very important for the clinical treatment of large bone defects in long bones,laying the foundation for clinical treatment.

Objective Articular cartilage injury is one of the most common orthopedic diseases with high morbidity and morbidity,especially in the elderly. Articular cartilage injury causes degenerative changes of articular cartilage, such as osteoarthritis, which can lead to disability, pain during joint movement and deformation of bone and joint. The prevalence of osteoarthritis accounts for 10% ～12% of the total population in the world. It is a common disease. The prevalence of osteoarthritis has increased to 49. 7% for the elderly aged over 65 years old ( Statistics of the World Health Organization ( who) in 2010 show that with the development of social aging and obesity and other adverse factors,these figures will continue to rise. It is known that osteoarthritis is related to aging,trauma,genetic susceptibility,obesity and inflammation,but the specific cause of osteoarthritis has not been fully identified, which leads to many obstacles in clinical treatment of osteoarthritis. At present,most of the clinical and research work in this field is focused on the restoration of cartilage trauma. In this review, we summarize and discuss the methods of cartilage defect repair,as well as the hot spots and directions of future research work.

Objective To discuss the effectiveness and safety of Ommaya sar implantation through neuronavigation in treatment of intracranial cysts. Methods Twenty-eight patients with intracranial cysts (cystic glioma, cystic metastases, radioactive encephalopathy cystic necrosis), admitted to our hospital from January 2007 to December 2014, were chosen in our study; these patients accepted Ommaya sar implantation through neuronavigation. The clinical data, disease courses, CT scan results, operation efficacies and postoperative complications of these patients were retrospectively analyzed. Results In the 8 patients with cystic gliomas, improvement of clinical symptoms and activity of daily living was noted in 6 patients; imaging re-check indicated focus shrink<50%in 6 patients, enjoying an effective rate of 75%. In the 12 patients with cystic metastases, the improvement of clinical symptoms and activity of daily living was noted in 9 patients; imaging re-check indicated focus shrink<50% in 9 patients, enjoying an effective rate of 75%. In the 8 patients with radioactive encephalopathy cystic necrosis, the improvement of clinical symptoms and activity of daily living was noted in 7 patients;imaging re-check indicated focus shrink<50%in 7 patients, enjoying an effective rate of 87.5%. No antidromic intracranial infection was noted in these 28 patients. Conclusion Implantation of Ommaya sar through neuronavigation is an effective treatment in intracranial cysts, enjoying minimally invasiveness.

Objective To investigate the effect of exosomes secreted from human lung adenocarcinoma A549 cells under hypoxic or normoxic conditions on the radiosensitivity and invasiveness of normoxia cells.Methods A549 cells were cultured in hypoxic (1％ O2) and normoxic (21％ O2) conditions,respectively.The exosomes (N-EXO and H-EXO) secreted from normoxic or hypoxic A549 cells were collected by ultracentrifugation and its number was measured using a NanoSight detector.The appearance and size distribution of exosomes were observed by a scanning electron microscopy.The exosomal marker protein CD63 was measured by Western blot.The proliferation of cells exposed to X-rays under hypoxic or normoxic conditions were detected by CCK8 assay.The cell uptake situation of exosomes labeled with PKH67 was observed by a fluorescence microscopy.Cell migration and invasiveness were detected by a cell scratch test and transwell assay.The expression of matrix metalloproteinase 2 (MMP2) and MMP9 was detected by ELISA.Cellular radioresistance effect of exosomes was evaluated by a colony formation assay.Results The NanoSight measurement showed the number of exosomes in cell culture medium was increased after hypoxia treatment.The H-EXO and N-EXO showed typical ring cake shape.The size distribution of H-EXO was mainly between 30 nm and 200 nm,smaller than that of N-EXO (50-220 nm).Western blot assay showed that CD63 was expressed in both H-EXO and N-EXO.At 4 and 6 days after 2 Gy X-rays irradiation,cell proliferation rate of hypoxia A549 cells was significantly higher than that of normoxia cells.The green fluorescent marker of exosomes,PKH67,was distributed inside of the cell.Cell scratch test showed that the width of H-EXO group was much smaller than that of N-EXO group at 12,24 and 48 hours after exosomes treatment (t =2.96,6.76,3.35,P ＜ 0.05).Transwell assay showed that the number of transmembrane cells in the H-EXO group was more than that in the N-EXO group and the control group (t =4.84,7.88,P ＜ 0.O1).The expression levels of MMP2 (t =4.70,3.21,P＜0.05) and MMP9 (t =5.61,3.76,P＜0.05) in the supernatant of H-EXO group were significantly higher than those in the control and N-EXO groups.Cell survival assay showed that the D0 values of control,N-EXO and H-EXO group were 2.614,2.552 and 4.50 respectively,indicating that H-EXO could enhance radioresistance of A549 cells significantly.Conclusions This study finds that the number of exosomes released from A549 cells was increased under hypoxic condition but its size becomes smaller than that under normoxia.Hypoxic exosomes can promote the migration of normoxia cells andenhance cell radioresistance as well.