Objective To explore the effect of neural cell adhesion molecule (NCAM) on adhesion,migration and morphology of mouse bone marrow-derived mesenchymal stem cells (BMSCs).Methods We isolated and cultured BMSCs from wild-type and NCAM gene knockout mice.The expression of NCAM was detected by Western blot and immunofluorescence.Wound healing and adhesion assays were used to detect cell migration and adhesion ability respectively.The morphological changes were observed and the expressings of protein β1 integrin,E-cadherin,β-catenin and N-cadherin were analysed by Western blot.Results The migration and adhesion of BMSCs were significantly reduced after NCAM gene knockout.Meanwhile,the expression of β1 integrin was lower than those in wild-type BMSCs (P<0.01).The morphology of NCAM gene knockout BMSCs changed from irregular to flattened,and expressed epithelial identification marker E-cadherin and β-catenin (P<0.05).However,the expression level of mesenchymal identification marker N-cadherin was decreased (P<0.01).Conclusions NCAM is involved in adhesion and migration of BMSCs via regulating the expression of β1 integrin.Furthermore,NCAMmay negatively regulate the mesenchymal-epithelial transitions of BMSCs.

HSP90, which is the biomarker of cell stress and endogenous protective protein, functions as a molecular chaperone. Many client proteins of HSP90, including EGFR, Met, Raf-1, IKK and p53, play important roles in the occurrence and development of tumor. Binding of HSP90 inhibitors triggers the deactivation of HSP90, resulting in client protein degradation, and hence inhibits the tumor growth by blocking multiple targets involved in signaling of tumor proliferation. This review summarizes recent development of small molecule inhibitors bound to N-terminal of HSP90.
Antineoplastic Agents ; chemistry ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Neoplasms ; Signal Transduction

OBJECTIVETo investigate the early clinical efficacy of induced membrane technique for reconstruction of large bone defects after debridement in adults with chronic osteomyelitis of limbs.

METHODSFrom March 2010 to March 2012,a total of 23 adult patients with chronic osteomyelitis of limbs were treated in our department. There were 15 males and 8 females, with a mean age 35.2 years old (ranged from 26 to 49 years old). Sixteen patients had open fracture history. According to the lesion site, there were 12 cases of tibia, 7 cases of femur, 3 cases of humerus, and 1 case of both radius and ulna. Among them, 19 patients had diseases in diaphysis and 4 patients in the metaphysis. The mean interval from infection to operation was 6.9 months (ranged from 4 to 13 months). All the patients were treated by using induced membrane technique. The follow-up evaluation included clinical complications, time of bone healing and limbs function. The Chinese version of SF-36 scores was used in the assessment of quality of life pre- and post-operation.

RESULTSThe average duration of follow-up was (27.6 ± 5.3) months (ranged from 18 to 43 months). Two patients had postoperative flap edge necrosis, 1 patient had superficial iliac incision infection, no obvious complications were recorded. Twenty patients obtained radiological union at a mean time of 4.6 months (ranged from 3 to 7 months). Among them, 16 patients treated with lower limbs surgery achieved full weight-bearing at about 5.2 months (ranged from 4 to 8 months) postoperatively. Four patients suffered from reinfection during follow-up, but 3 of them achieved complete bone healing after the second surgeries with induced membrane technique. At the final follow-up, there was a substantial improvement in each dimension scores and total scores of SF-36 as compared with those before surgery.

CONCLUSIONWhen treating with adult chronic osteomyelitis of limbs, the induced membrane technique can effectively reconstruct large bone defects after debridement, significantly shorten treatment cycle, provide satisfactory results with minimal complications, promote good recovery of limbs function and require relatively simple operation technique.

Adult ; Chronic Disease ; Extremities ; surgery ; Female ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery ; Reconstructive Surgical Procedures ; methods

An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.

OBJECTIVETo evaluate the early clinical and radiographic outcome of scaphoid non-unions treated with Acutrak headless compression screw.

METHODSFrom January 2008 to July 2011,21 patients with scaphoid non-union were treated in our department. There were 18 males and 3 females with a mean age of (23.6 +/- 4.6) years; 12 cases were on right hand and 9 were on left. According to Herbert-Fisher classification, there were 10 cases with type D1, 7 cases with type D2, 3 cases with type D3, and 1 case with type D4. The mean time from injury to operation was (12.4 +/- 2.7) months. All patients were treated with Acutrak headless compression screw fixation (6 cases received 2 screws fixation, 15 cases received 1 screw fixation, and Matti-Russe bone grafting was applied in 7 cases). The carpal height, the scaphoid index and changes of the scapholunate angle were assessed before and after the operation. Range of motion and grip strength were recorded and the wrist function was assessed according to the Patient-Rated Wrist Evaluation (PRWE).

RESULTSAverage duration of follow-up was (21.3 +/- 3.6) months. All the patients attained radiological union in a mean time of (13.3 +/- 2.4) weeks following the operation. No obvious complications were recorded. The surgical treatment allowed the preoperative mean scaphoid index of 0.61 +/- 0.13 and the preoperative mean scapholunate angle of (59.4 +/- 6.8) degree to be improved to 0.69 +/- 0.10 and (44.3 +/- 8.2)degree postoperatively, respectively. There was a substantial improvement in grip strength and pain amelioration after surgery. The preoperative mean PRWE score of 45.2 +/- 4.7 was improved to 76.1 +/- 5.2 postoperatively. All patients returned back to the original work,the average time from surgery to work was (6.0 +/- 1.1) months.

CONCLUSIONFor scaphoid non-unions, Acutrak headless compression screw fixation can provide anatomical reduction, provide satisfactory results with a high union rate, well return of function and minimal complications in the early stage.

Adolescent ; Adult ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Ununited ; surgery ; Humans ; Male ; Range of Motion, Articular ; Scaphoid Bone ; injuries ; physiopathology ; surgery ; Treatment Outcome ; Wrist Injuries ; physiopathology ; surgery ; Wrist Joint ; physiopathology ; surgery ; Young Adult

It has been received wide attention for the increasing prevalence of myopia worldwide in recent years.The myopic fundus lesions,which may be associated with the variation level of the thickness of retinal and choroidal membrane in the early stage of myopia,remained the main cause of vision impairment and reduce the quality of life.However,the changes in retinal and choroidal thickness in the progress of myopia and the associated factors are not clear.This article gives a brief review the change in the thickness of retinal and choroidal thickness in progress of myopia and its associated factors,which can provide support for clinical diagnosis,prevention and monitoring of macula/choroidal associated diseases.

Objective To establish the chick embryo chorioallantioc membrane(CAM)as a model for in vivo research on endometriosis.The model was used to investigate the mechanism of anti-vascular endothelial growth factor(VEGF)antibody for treatment of endometriosis.Methods Human endometrial fragments were explanted onto the CAM.Then anti-VEGF antibody was used for the endometriosis-like lesions after transplantation of human endometrial fragments.The CAM models were treated respectively as control groups and experimental groups.The terminal deoxynucleotidyl transferase-mediated biotin- deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL),proliferating cell nuclear antigen(PCNA) and microvessel density(MVD)were used in vivo for analysis of anti-angiogenesis.Results The apoptosis intensity of anti-VEGF antibody treated groups(6.7?0,9,6.9?0.8)was significantly higher than that of the control groups(5.0?0.9,5.4?1.1;P

This study was aimed to investigate the expression level of perforin in cord blood NK cells and the relation of perforin expression after IL-2, IL-15 stimulation to cytotoxicity of NK cells. NK cells were isolated from cord blood MNC by depleting CD3(+) cells and then enriching CD56(+) cells using immunomagnetic separation (CD3 and CD56 cell isolation kit, autoMACS, miltenyi). The purity was analysed by flow cytometry. According to the different combination of cytokines, there were two groups: group A (IL-2) and group B (IL-2 + IL-15). The cytotoxicity and perforin expression rate of fresh and different cultured CB-NK cells against K562/Jurkat cell lines were estimated by LDH release test (cytotoxic 96 non-radioactive cytotoxicity assay). The results showed that the purity of NK cells after separation was more than 90%. The cytotoxicity towards both tumor lines in group B was higher than that in group A (p < 0.05), and cytotoxicity in group A was higher than that of fresh NK cells (p < 0.05). Perforin expression rate of group A (84.55%) was higher than that of fresh NK cells (67.21%) (p < 0.05), and there was no significant difference between group A and B (84.55% versus 87.22%) Cytotoxic activity of CB-NK cells was positively correlated with perforin expression rate (r = 0.886, p < 0.05). It is concluded that IL-2 can enhance cytotoxicity of CB/BM-NK cells by increasing the perforin expression.
CD56 Antigen ; metabolism ; Cells, Cultured ; Cytotoxicity, Immunologic ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; metabolism ; Perforin ; metabolism

The aim of this study was to explore the cytotoxicity of fresh cord blood(CB) NK cells and the influence of IL-12 and IL-15 on activity of the NK cells killing K562 and Jurkat cells lines. The NK cells were isolated from cord blood by depleting CD3(+) cells and then enriching CD56(+) cells using sorting with immunomagnetic beads. The experiment was divided into 3 groups: group A (fresh CB-NK cells without cytokines), group B (CB-NK cells cultured by IL-2) and group C (CB-NK cells cultured by IL-2 and IL-15). The purity of NK cells was determined by flow cytometry; the cytotoxity of fresh and different cytokine-treated CB-NK cells on K562 and Jurkat cell lines was detected by LDH release test. The results showed that the purity of NK cells before and after sorting was 14.88 ± 9.2% and 92.39 ± 0.8% respectively. After culture for 3 days, NK-forming colony amounts in group B and group C were 148.60 ± 13.0 and 831.80 ± 23.0 respectively, the comparison between group B and group C showed the significant difference (p < 0.05). The cytotoxicities of NK cells in group A, B and C on K562 and Jurkat cell lines were 27.76 ± 8.8%, 61.90 ± 9.1% and 87.62 ± 3.7%; 29.32 ± 2.5%, 69.43 ± 4.4% and 92.95 ± 3.2% respectively, the difference was significant (p < 0.05). It is concluded that the fresh isolated CB-NK cells show low cytotoxic activity. After stimulated with IL-2 or IL-2 plus IL15, cytotoxicity of CB-NK cells increases obviously, the effect of IL-2 plus IL-15 is much better than IL-2 alone for promoting the growth and enhancing the cytotoxicity of CB-NK cells.
Cytotoxicity, Immunologic ; drug effects ; immunology ; Fetal Blood ; drug effects ; immunology ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Jurkat Cells ; K562 Cells ; Killer Cells, Natural ; drug effects ; immunology

OBJECTIVETo compare and evaluate the biocompatibility of three kinds of dentin bonding agents Xeno III (XO), Adper Prompt (AP), Single bond2 (SB) through cell culture in vitro.

METHODSThree kinds of dentin bonding agents (XO, AP, SB) were applied on the surface of the dental slices which were 5.0 mm in diameter and 0.5 mm in depth. By immersing the slices into the DMEM culture medium, the maceration extracts were obtained. Normal dental pulps of teenagers were collected and human pulp fibroblast was cultured using tissue explant method. The fifth generation pulp cells were exposed to culture medium containing different concentrations of maceration extracts (100.0%, 50.0%, 25.0%, 12.5%) for 24, 72, 120 h. At last, MTT method was used to evaluate the cytotoxicity of the dentin bonding agents on human pulp fibroblast.

RESULTSThe results showed that all three kinds of dentin bonding systems had cytotoxicity to human pulp fibroblast in different degree in vitro. The cytotoxicity of XO and AP was less than SB. The difference was statistically significant (P<0.05).

CONCLUSIONThe results of cell culture in vitro indicated that total-etching adhesives system has more irritation to pulp than self-etching adhesives system.

Adhesives ; Adolescent ; Dental Pulp ; Dentin ; Dentin-Bonding Agents ; Fibroblasts ; Humans ; Resin Cements