AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.

The basical structure of diketopiperazines is a cyclic dipeptide condensed by two amino acids. Because of the stable framework of the six-member ring structure, and having two hydrogen bond donor and two hydrogen bond receptor, DKPs have become important chemical pharmacophores, with strong biological activities and pharmacological activities in the drug. A series of cyclic compounds were found from marine organisms in recent years, research showed that their functions are not limited on anti-bacterial, cytotoxic activity, and so on, but also playing an important role in regulatory mechanism of quorum sensing as signal molecules, they have become research hot point in ecological chemistry. This paper reviewed the research progress of diketopiperazines found in the marine microbial metabolites, and the future study trends was discussed and outlooked.

Objective:To investigate the value of the three-dimensional CT reconstruction by computer photo process system for laryngeal carcinoma and control study with pathologic feature. Methods: Fifty patients with laryngeal carcinoma were scanned by helical CT which photos were three-dimensionally reconstructed with a photo processing system,and the images were compared with pathologic feature.Results:Clear three-dimensional photo of the laryngeal was obtained through the reconstruction of the helical CT.The images could be circular fully around the axis of X,Y and Z in order to find the best view of the diagnosis.Ninety-six percentage of the TNM stage of patients according to the images of three-dimensional reconstruction were consistent with the TNM stage after operation.The safe edge was obtained in 98% of the surgery for patients.Good correlations were showed in the images of the three-dimensional reconstruction of the larynx and pathologic specimens.Conclusion:Good consistent of the three-dimensional images of the larynx with pathologic specimen were useful for diagnosis,operation design and education.

Objective To retrospectively analyze clinical features,treatment methods and efficacy of giant cell tumor of bone in proximal tibia,and to investigate risk factors affecting tumor recurrence and functional outcomes.Methods A total of 250 patients with giant cell tumor of bone in proximal tibia confirmed by pathology,who had undergone surgical treatment from March 2000 to July 2014,were enrolled in this study.There were 132 males and 118 females,with an average age of (34.59±12.86) years.A total of 140 patients who were followed up for more than 3 years were included in this study,and there were 72 males and 68 females,with an average age of (34.46± 11.96) years.There were 11 cases of Campanacci grade Ⅰ,58 cases of grade Ⅱ,71 cases of grade Ⅲ and pathological fracture of 47 cases.According to surgical methods,they were divided into bone grafting group (49 cases),bone cement filling group (34 cases),prosthesis group (46 cases) and others group (11 cases).The epidemiology,clinical and radiographic features and risk factors affecting tumor recurrence and functional outcomes were analyzed.Results A total of 140 patients were followed up,the follow-up period was 36-324 months,with an average of 95.4 months,and the median follow-up time was 88 months.Recurrence was found in 26 cases,and recurrence rate was 18.57％,with an average recurrence interval of 25.85 months.Recurrence was found in 17 cases in the first 2 years.The 5-year free survival rate was 77.60％.The recurrence rates were 18.37％ in bone grafting group,20.59％ in bone cement filling group,15.22％ in prosthesis group and 27.27％ in the others group,no statistically difference was found on recurrence rate and free survival rate (P=0.805,P=0.558).Recurrence was not related to all kinds of factors.A variety of related factors affecting postoperative recurrence were analyzed,sex,the first diagnosis of the original recurrence,left and right side,whether the eccentricity,fracture,cortical bone destruction,soft tissue mass,surgical methods,high-speed grinding,auxiliary application,and there was no significant correlation between recurrence and these factors.The MSTS 93 score was 25.26±4.31.Function of the primary patients was better than that of recurrence (P=0.044).Function of the patients treated with curettage with or without internal fixation was better than that with segmental resection (P=0.011).Function of the patients treated with grafting or bone cement filling was better than that with prosthesis or allograft-prosthesis reconstruction (P=0.004).There were no significant correlation between MSTS function score and gender,left and right side,whether the eccentricity,whether fractures,cortical bone destruction (Campanacci grade),whether there is soft tissue mass,whether the use of assisted inactivation,whether the use of grinding or internal fixation.Conclusion Various surgical methods had no significant effect on the recurrence of proximal tibial GCT,as for GCT in proximal tibia,there is no relation between recucrrence and related factors.Whether primary tumor and surgical methods are two important factors affecting limb function.

Denosumab is a fully human monoclonal antibody on receptor activator of NF-κB ligand that has shown adjuvant treatment for giant cell tumour of bone (GCT).Clinical trials show that over 85％ of patients have significantly improved their clinical symptoms,imaging and histology.Currently mainly used for central axis bone such as the sacrum and spine of difficult surgical excision or limb recurrence or refractory GCT patients.Case reports have demonstrated complete response or tumor stabilization with denosumab,allowing for surgical procedures in simplify.However,the duration of the medication and the optimal therapeutic dose and long-term effects are not yet known.The local high recurrence rate after discontinuation of the drug remains a problem with the accumulation of clinical research,and the follow-up time is prolonged.More noteworthy is the possibility of pseudosarcoma,even malignant transformation after Denosumab treatment.This sarcoma transformation requires further controlled studies and long-term of follow-up to reach a definitive conclusion.In this paper,we retrospective analysis of the application status,imaging and histology related research of Denosumab in the treatment of GCT.Correct understanding of the value and clinical significance of this drug in GCT treatment requires a multi-center study and a long-term follow-up to evaluate the clinical value.

Objective To analyze clinical efficacy of artificial prosthesis in giant cell tumor in distal femur,and to investigate risk factors affecting prosthesis failure and functional outcomes.Methods 42 patients with giant cell tumor of bone in distal femur,who had undergo prosthesis replacement from January 2002 to May 2015,were enrolled in this study.There were 24 males and 18 females,with an average age of 38.53± 12.87 years.There were 28 primary patients and 14 relapsed patients,including 11 cases of recurrence once and 3 cases of twice.Three-dimensional finite element model was used to analyze the effect of different angles of deviation of the spinal needle on the deformation of the bone wall.The correlations between the factors such as age,sex,occupation,prosthesis type,and other factors on prosthesis loosening were compared.Biomechanical effect of lower limbs caused by prosthesis offset angle was analyzed through gait analysis.Analyzed the effects of primary tumor or recurrence,prosthesis service status,and length of surgical osteotomy on joint function.Results A total of 42 patients were followed up by 20-158 months,with an average of 68.7 months.The 3 year survival rate of prosthesis was 83.33％ for 3 years and 57.14％ for 5 years.The major reason of prosthesis failure was loose (18/42,42.8％).X-ray films showed 19 cases of prosthetic intramedullary nail and sagittal bias of medullary force line angle ＞ 3° in the first follow-up.Osteotomy length (OR=0.132,P=0.0027) and offset angle of needle (OR=25.000,P=0.000) were significantly correlated to prosthesis loose.A length more than 12 cm and angle more than 3° were easier to result in prosthesis loose.There were no significant correlation between prosthesis failure and patients age,gender,occupation and prosthesis type.Gait analysis shows that the unsuitable bias angle of the prosthesis can significantly change the joint force of the prosthesis.The average score of MSTS 93 function evaluation was 25.43±4.256,excellent in 33 cases,good in 7 cases and poor in 2 cases.Function of patients with primary GCT were better than that of recurrent ones.Patients with one 1 times recurrence were better than that of recurrence twice (P=0.003).Patients without prosthesis loosening and revision were better than that with loosening (P=0.001).Patients with an osteotomy length less than 12 cm had a poorer function than that with more than 12 cm (P=0.002).Conclusion The main factors affecting distal femoral prosthesis replacement therapy of GCT is loosening,which was caused by broach and medullary cavity mismatch,osteotomy length,prosthesis rotation,prosthesis position.The function of the prosthesis is mainly affected by operation times,prosthesis status,osteotomy length and low patella.

Objective:To investigate the prognostic value of molecular subtypes in patients with resected invasive breast cancer.Methods:Between 2015 and 2018 7 869 patients with invasive breast cancer after undergoing surgery were included in this analysis. Breast cancer was classified into four subtypes according to the status of hormone receptor (HR) and HER2: HR+/HER2-, HR+/HER2+, HR-/HER2+, and HR-/HER2-. Kaplan-Meier curves and COX regression were used to compare disease-free survival (DFS) and overall survival (OS) among different subtypes.Results:The 5-year DFS and OS were 86.30% and 94.29%, respectively. Proportions of HR+/HER2-、HR+/HER2+、HR-/HER2+ and HR-/HER2- were 52.9%、17.5%、14.1%和15.5%, respectively. The 5-year DFS of HR+/HER2- subtype (88.12%) was higher than HR+/HER2+ (84.67%, P=0.026), HR-/HER2+ (84.19%, P<0.001) and HR-/HER2- (83.70%, P<0.001). The 5-year OS of HR+/HER2- (95.38%) was not different from HR+/HER2+ (95.17%, P=0.187), while it was higher than that of HR-/HER2+ (92.26%, P<0.001) and HR-/HER2- (91.69%, P<0.001). Subtype was still a significant factor regarding DFS and OS in multivariable analyses adjusting for age, sex, stage, Ki67, types and time of surgery. The DFS ( P=0.257) and OS ( P=0.511) was not different between HR-/HER2+与HR+/HER2- subtypes, while HR-/HER2+ and HR-/HER2- patients had worse DFS ( P<0.05) and OS ( P<0.05) than that with HR+/HER2-. Conclusions:Molecular subtype is a significant independent prognostic factor for DFS and OS in operable invasive breast cancer. HR+ subtypes have better prognosis compared with HR- subtypes. The DFS and OS were not different between HR+/HER2- and HR+/HER2+, or between HR-/HER2+ and HR-/HER2-.