Objective To clone,express and identify the nuclear antigen Sm B′in E. coli to establish a new assay for detecting autoanti-body to Sm B′. Methods A full length cDNA of Sm B′was cloned from cell line HL-60 by RT-PCR. The PCR product was TA cloned and sequenced and inserted into the vector pGEX-5T. The recombinant plasmid was transformed into E. coli BL21. The positive clones were identified by restricted enzymes and induced by IPTG. The expression product was analyzed by SDS-PAGE and Western blot. Results The PCR product was about 700 bp in size which was in accordance with predicted 657 bp and sequencing result showed consistent with the sequence in GenBank. The pGEX-5T-Sm B′positive clone produced a 51 000 kD of fusion protein which was immunoreac-tive with anti-Sin B′confirmed by SDS-PAGE and Western blot. Conclusion The successful cloning and expression of nuclear antigen Sm B′laid a foundation for further research work.

Abnormal metabolism in vivo of uric acid leads to many diseases.Recent discovery shows that glucose transporter-like protein-9,which belongs to glucose transporter family not only transports glucose,but also plays an important role in the process of uric acid transport,which is also affected by pH,glucose,estrogen,etc.The variation of glucose transporter-like protein-9 results in metabolic diseases.Therefore,the study of glucose transporterlike protein-9 in uric acid transport and related drug will provide new ideas to control the development of hyperuricemia and related cardiovascular disease.

Objective To establish a new assay for detecting autoantibody Jo-1, cloning and expressing human autoantigen Jo-1 in E.coli.Methods A full length cDNA of human autoantigen Jo-1 was cloned from cell line HL-60 by RT-PCR. The PCR product was TA cloned, sequenced and inserted into the carrier pGEX-5T.The recombinant plasmid was transformed into E.coli BL-21. The positive clones were identified by restricted enzymes and induced by IPTG. The expression product was analyzed by SDS-PAGE and Western blot.Results The PCR product was about 1 500 bp in size which was in accordance with predicted 1 526 bp and sequencing result showed the same with GenBank′s report.The pGEX-5T- Jo-1 positive clone produced a 75 000 fusion protein which had natural immunogenicity of human autoantigen Jo-1 by SDS-PAGE and Western blot.Conclusion Successfully cloning and expression of human autoantigen Jo-1 laid a foundation for further research work.

A kinetics model was developed for predicting and simulating immobilized cellulase performance, which follows Michaelis-Menten kinetics with competitive product inhibition. Taking into account the effects of competitive product inhibition, inner diffusional limitation, substrate concentration and carrier size, the substrate distribution and the product distribution in carriers were investigated, and the effectiveness factors were also calculated over a wide range of parameters. The effects of competitive product inhibition are shown to increase the substrate concentration in the carrier, and, additionally, to increase the effectiveness factors slightly. With the increase of inner diffusion coefficient, both the effectiveness factors and the substrate concentration in the carrier increase. As the carrier size increases, on the other hand, these values decrease. The effectiveness factors and the substrate concentration in the carrier are found to increase when substrate concentration in the reaction system increases.
Cellulase ; metabolism ; Diffusion ; Enzymes, Immobilized ; metabolism ; Kinetics ; Microspheres ; Models, Chemical ; Particle Size ; Substrate Specificity

Objective To elucidate the immunologic mechanism and clinical effect of high intensity focused ultrasound (HIFU) combined with dendritic cell and cytokine induced killer cell (DC-CIK) immunotherapy on patients with pancreatic cancer.Methods Seventy-two pancreatic cancer patients were divided randomly into 2 equal groups,one treated with HIFU only the other treated with HIFU and DC-CIK immunotherapy.Ultrasound imaging and a variety of immunological indexes were recorded before and after treatment and the clinical effects in the two groups were compared.Moreover,autogenous tumor cells were isolated from the combination therapy group and the killing activity of DC-CIK which loaded tumor antigen processed by HIFU on autogenous tumor cells was observed.Results Tumor antigen processed by HIFU can improve the killing activity of DC-CIK on autogenous tumor cells.After treatment,the immunological indexes,of all patients were better than before treatment.(58.26 ± 17.97 versus 52.15 ± 14.22 pg/ml with IL-12 22.14 ± 6.39 versus 17.36 ± 5.73 ng/ml with HSP70 and 0.94 ± O.34 versus 1.32 ± O.61 ng/ml with TGF-β,P ＜ 0.05 ) ; The combination group was significantly better than the HIFU group with regard to the average scores of quality of life (75.89 ± 19.65 versus 67.22 ± 16.34,P＜0.05),pain (3.15 ±0.82 versus 3.59 ± 1.04,P ＜0.05),tumor markers (107.55 ±27.58 versus 123.63 ±34.12 U/ml) and survival time (18.92±6.47 versus 13.36 ±5.78 mos).Conclusion HIFU can improve the immunologic status and anti-tumor response in patients with pancreatic cancer.HIFU combined with DC-CIK has good synergistic therapeutic effect for treating pancreatic cancer.

PBL (Problem-based learning) is a kind of teaching method, which is helpful for guiding students to solve practical and complex problems. The compiling of PBL teaching plans is the key to the implementation of PBL teaching. According to the teaching outline of internal medicine, we set the PBL teaching plan, formulated the teaching goal, collected the clinical real cases, finished the compiling of PBL cases of student version and teacher version, and preceded teaching practice accord-ing to the PBL cases. Improving and constructing the compiling framework of internal medicine PBL teaching plan laid the theoretical foundation for carrying out internal medicine PBL teaching.

The improvement of diagnostics teaching system,including the establishment of curriculum system and evaluation system,is the base of promoting clinical- medicine teaching.Our study showed that the theoretical knowledge and clinical skill of medical students could be improved by constructing clinical diagnostics curriculum system and improving organization management and assessment system,which could pave the way for the transition from medical students to clinicians.

Objective:To investigate the relationship between the distance from the upper and lower resection margin and the gastric cancer patients with R0 resection and no distant metastasis.Methods:Retrospective analysis of 281 patients with gastric cancer in our hospital,the relationship between the age,sex,tumor size,tumor size,vascular invasion,lymph node metastasis,TNM stage,type of gastric cancer,tumor location and operation mode was analyzed.To compare the survival time of patients with different upper and lower margins.Results:The increase of the distance from upper resection margin was significantly related to the tumor size＞5 cm,TNM stage,type of gastric cancer,tumor location,and the difference was statistically significant(P＜0.05);The distance of lower resection margin was significantly correlated with tumor size＞5cm,vascular invasion,lymph node metastasis and TNM stage,type of gastric cancer,and the difference was statistically significant (P＜0.05);Themedian survival timein patientsof upper resection marginr□3cm with 48 months of the 5-year follow-up period was significantly higher than that in patients of the resection margin＞3cmwith 46 months (P＜0.001).Themedian survival timein patientsof lower resection margin□3cm with 45 months of the 5-year follow-up period was significantly higher than that in patients of the resection margin＞3cm with 44 months (P＜0.001).Conclusion:Gastric cancer postoperative upper and lower resection margin was significantly related with tumor size,TNM staging factors,and the median survival time of upper and lower resection margin＞5 cm was significantly lower than that of the resection margin3 cm.

Actins ; metabolism ; Adult ; Amputation ; Arthroplasty, Replacement, Knee ; Bone Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Calmodulin-Binding Proteins ; metabolism ; Humans ; Leiomyosarcoma ; diagnosis ; metabolism ; pathology ; surgery ; Magnetic Resonance Imaging ; Male ; Radiography ; Tibia ; diagnostic imaging ; surgery

Objective To construct stable chromokinesin KIF4A knockdown gastric cancer cell lines (SGC-7901) and study the mitotic spindle midzone formation in these cells.Methods Short hairpin RNA (shRNA) expression plasmids of pGPU-GFP-shKIF4A,pGPU-GFP-shNC specific targetting KIF4A and non-sense DNA sequences were constructed respectively and then transfected into SGC-7901 cells.The cells were cultured in selection medium containing G418 to form single clones.Stable KIF4A shRNA expressing SGC-7901 cells were picked up under an fluoroscent microscope and identified by Western Blot.The spindle midzone in these cells was analyzed after immunofluorescence staining.Results Three cell lines stably expressing KIF4A shRNA and one with shNC were successfully constructed.Compare to the control cell line,KIF4A knockdown cell lines represented remarkable elongation of spindle midzone and the less the KIF4A,the longer spindle midzone.Conclusions Stable KIF4A knockdown SGC-7901 cell lines were successfully constructed that can serve for further study of KIF4A ' s function and its role in proliferation and development of gastric cancer.