Objective　To investigate the pattern of human neutrophil degranulation mediated by Fc alpha receptors. Methods　IgA immune complexes (IgA IC) and specific monoclonal antibody were used to stimulate neutrophil degranulation. Lactoferrin released by stimulated neutrophils was quantitatively determined. Results　Monoclonal antibody and IgA IC could both induce neutrophil lactoferrin release. However, monoclonal antibody induced rapid and strong degranulation whilst the effect of IgA IC was much weaker. Conclusion　The pattern of neutrophil degranulation induced by IgA IC and monoclonal antibody is different.

Objective To establish more simple and effective rat limbs transplantation. Methods Taking Wister rat as donatur,SD rat as recipient,and transplanting Wister rat rear limbs (right or left) to SD rat.The experiments were carried out in two group,the traditional group (after mutilation of SD rat limbs, transplanting the similar side Wister rat rear limbs to SD rat by fixing bone,anastomosing nervi vasorum, suturing muscle and skin) and the improvement group (retaining SD rat limbs,and fixing Wister right limbs on the left inner side of SD rat limbs or fixing Wister left limbs on the right inner side of SD rat limbs by suturing muscle and skin. Then only anastomoing femoral artery and femoral vein, and not anastoming nervi, not fixing bone). Recording operation time and weight change after operation. Microsurgery usual care Recording host drink and food and weight. Observing implant change. Results Seventy-four operations were accomplished successfully. Graft rejection reappear stably. All 74 rats survived over 14 days after transplantation. The operation time of traditional group and imrovement group were (125±40) min and (70 ±21) min respectively. Decreased body weight of traditional group and improvement group were (3.78± 1.09)and(2.05±0.90) g respectively. After 3 days, the weight of improvement group rat begin to increase, however, traditional group rat always dcrease in one week. Operation achievement ratio of traditional group and improvement group were 38%, 90% respectively. Improvement group is better than traditional group in above 4 section. Conclusion The method is more simple,applicable and requires shorter time. Less trauma to rats and rats recover quickly. It can be used in establishing rat limbs transplantation acute rejection model.

Objective To probe the surgical approach and effect on upper abdominal malignant tumor infiltrating pancrea. Methods Thirty patients with upper abdominal malignant tumor infiltrating pancreas or tissue around pancrea were treated by radical resection on primary tumor combined pancreaticoduodenectomy. Results Three patients died in 1 month after operation due to multiple organ failure. The patients with gallbladder cancer and metastatic lymph nodes fixed behind caput pancreatis were followed up averaged 35 months. The patients with gastric antrum carcinoma infiltrating caput pancreatis were followed up averaged 31 months. The patients with recurrent gastric cancer infiltrating caput pancreatis were followed up averaged 13 months. The patients with pancreatic and duodenal invasion by cancer of hepatic flexure of colon were followed up averaged 41 months. The patients with hilar cholangiocarcinoma and metastatic lymph nodes fixed behind caput pancreatis were followed up averaged 11 months. Conclusion The patients with upper abdominal malignant tumor infiltrating pancreas should be operated by radical resection on primary tumor combined pancreaticoduodenectomy, which can increase the rate of tumor resection, and be expected to prolong the survival period.

Objective To evaluate the application of measurement of T2*value,width of substantia nigra pars compacta (SNc) and the ratio of the width to the midbrain diameter in diagnosing Parkinson disease (PD) in early stage with susceptibility weighted imaging ( SWI) by 3T MR. Methods 59 patients with early stage idiopathic PD patients and 59 healthy controls,ranging in same ages and gender,had been scanned with routine sequences and SWI sequences by 3T MR. T2*value,width and the ratio of the width to the midbrain diameter of SNc were measured. The results of measurement were analyzed and compared. Results (1) The T2*values, width and the ratio of the width to the midbrain diameter was decreased in homolateral side SNc of symptoms of subjects with PD compared with the healthy controls ( <0.05) . (2) There was a significant reduction in the T2*values and the ratio of the width to the midbrain diameter in contralateral side SNc symptoms of subjects with PD compared with the healthy controls (P<0.05) . There was no differences in width of SNc ( >0.05) . Conclusion Measurement of T2*value, width and the ratio of the width to the midbrain diameter of SNc with SWI is reliable to diagnose PD.

Objective To clone the aldehyde dehydrogenase (adhA) gene from Methylovorus glucosotrophus and study its expression,purification and enzymatic characteristics.Methods The adhA gene was amplified and cloned to the expression vector pTIG.The AdhA was successfully expressed with induction in Escherichia coli BL21(DE3).The enzymatic characteristics were investigated by AHMT,and AdhA was purified by Ni+ exchange chromatography.Results AdhA accounted for more than 50% of the total cell proteins,and the purity was about 95%.With methanol as the substrate,the optimal pH of AdhA was 7.0,while the optimal temperature was 30℃.The enzymatic activity of purified AdhA remained about 60% when stored at room temperature for 6 days.Conclusion AdhA from MP688 is expressed in vitro,and methanol is the optimal substrate among all the substrates investigated.

Objective To prepare and characterize specific monoclonal antibodies( McAbs) against the heavy chain of botulinum neurotoxin serotype A ( BoNT/AHc ) .Methods BALB/c mice were immunized with purified BoNT/AHc protein.After the fusion of mouse splenic cells with SP2/0 cells, hybridoma cell lines secreted McAbs against BoNT/AHc. The McAbs obtained were characterized by indirect ELISA, Western blotting and rapid isotypingassay before being used in ELISA to detect interaction sites in McAbs and BoNT/AHc preliminarily.Results Antigen protein BoNT/AHc of high purification was obtained.Four hybridoma cell lines secreting McAbs against BoNT/AHc were screened,named 1A4,3H3, 3H7 and 5H8,respectively.Their titers of McAbs were all above 3.0 ×103 .They were specifically combined with BoNT/AHc protein by Western blotting.The isotype of 1A4 and 3H7 was IgG1(Κ),that of 3H3 was IgM(Κ),and that of 5H8 was IgG2b(Κ).Additive ELISA showed that epitopes recognized by the four McAbs were close.ELISA analysis confirmed the interaction epitopes in McAbs and BoNT/AHc.Conclusion Monoclonal antibodies against BoNT/AHc are prepared and characterized,providing effective tools for studying the neutralizing antibody and antibody epitopes of BoNT/AHc.

Aged ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Hemangioendothelioma, Epithelioid ; pathology ; Humans

Objective To investigate the relationship between the expression of decay receptor 3 (DcR3) and the eliniealpathological parameters in human gastric carcinoma. Methods The expression of DcR3 was examined by RT-PCR in a series of 41 human primary gastric carcinomas and 41cases of normal tissue adjacent to tumor. Multiple clinical pathological factors were analyzed according to their relation with the expression of DcR3. Results The positive rate of expression of DcR3 was 56 %(23/41) in human gastric carcinoma. The expression of DcR3 in gastric carcinoma was significantly higher than that in normal tissues adjacent tumor. The expression of DcR3 was significantly correlated with different degrees of differentiation, lymph node metastasis and TNM staging (P <0.10), but there was no significant difference in DcR3 and other clinical pathological features such as tumor position and invasion depth (P>0.10). The multiple linear regression equation was Y=0.432-0.208X1+0.098X2+0,086X3. Conclusion DcR3 expression can be highly found in gastric carcinoma. The abnormal expression of DcR3 may promote tumorigenesis and progression. DcR3 may be important in evaluating the tumor differentiation, infiltration depth, lymph node metastasis and TNM staging of human gastric carcinoma.

Objective:To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs ) for regeneration of tissue inj uries and defects. Methods:Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently. Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2,B27,basic fibroblast growth factor (bFGF),and epidermal growth factor (EGF).The skin dermis derived spheroids (SDDSs)were collected and monolayer cultured in serum-containing medium.Finally,the cells were characterized by immunofluorescence staining and differentiation assays.Results:The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium. After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number. The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin. Under osteogenic,chondrogenic and adipogenic differentiation conditions,these cells were differentiated into the alizarin red,safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs. However,safranin O staining was weaker in the dermis derived cells than BMSCs. Conclusion:A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials,demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.

Objective To explore humam cytomegalovirus(HCMV) encoded microRNAs during latent infection in order to help study HCMV virology and latent infection mechanisms.Methods A model of HCMV latent infection via THP-1 cells infected with HCMV was constructed.Deep-sequencing was performed using high-resolution Solexa sequencing platform.The secondary structure of the newly sequenced miRNA was predicted by RNAFold bioinformatics software. Results HCMV encoded various miRNAs during latent infection, including miR-US25-2-3p, miR-US25-2-5p, miR-UL112, miR-US25-1, miR-UL22A and PC-5p-148467 with a predicted length of 25 bp, named hcmv-miR-US33as-5p.Conclusion HCMV can express many types of miRNAs during latent infection.