OBJECTIVETo identify and characterize an unknown microorganism causing contamination in several mammalian cell cultures.METHODSThis bacterium was identified by 16S rRNA sequencing and studied by DAPI and DiOC6 (3) staining, Gram staining, acid-fast staining, and electron microscopy. The isolated bacterium was also used to infect host cells to observe antibiotic effectiveness and its relationship with host cells.RESULTSThe 16S rRNA sequence analysis shows that this rod-shaped microorganism belongs to the family Caulobacteraceae, class Alphaproteobacteria, and was most closely related to Phenylobacterium zucineum HLK1T strain. The bacterium collected in the "swimming" stage was Gram staining negative, but Gram staining positive in the "sessile" stage. Under the electron microscope both flagellated and non-flagellated types were found. So far, no antibiotics were effective to inhibit this microorganism. The contamination with this bacterium frequently led to failed resuscitation of thawed cells. We found that the cells resuscitated with the used culture supernatants were increased in number by 3-4 folds as compared to those resuscitated with freshly prepared media.CONCLUSIONPhenylobacterium may have a dimorphic life cycle including a swimming stage and a sessile stalked stage.

Objective To establish a nerve stem cell model possessing the potentiality of differentiating intoγ-aminobutyric acid neuron-like cells(GABAergic-like cells),and provide eligible investigative vector for study on GABAergic neurons degenerative diseases. Methods Dibutyryl cyclic adenosine monophosphate (dbcAMP)was applied to induce human neuroblastoma SH-SY5Y cells,and the SH-SY5Y cells were divided into control group (0 mmol·L-1 dbcAMP),0.3 mmol·L-1 dbcAMP group,0.6 mmol·L-1 dbcAMP group,1.0 mmol·L-1 dbcAMP group and 2.0 mmol·L-1 dbcAMP group;the morphological changes of SH-SY5Y cells were observed.The Image-Pro Plus 5.0 software was used to measure the length of neurites of the neuron-like cells, and the percentage of the SH-SY5Y cells with neurites longer than 30 μm was calculated. The immunofluorescence cytochemistry technique was utilized to test GAD65 positive cells,and the positive rate of immune response was calculated.Results The results of light microscope observation showed that the cells in control group were polygonal,circular or shuttle type with smooth membrane and clear boundaries. With the increasing of the concentrations of dbcAMP and the prolongation of time,the morphology of SH-SY5Y cells’bodies became smaller with longer processes in dbcAMP groups.The cells interwined each other and showed mature neuron phenotype in 1.0 mmol·L-1 dbcAMP group.Compared with control group(31.4%±4.2%),the percentages of the cell with neurites longer than 30 μm in 0.3, 0.6, 1.0, and 2.0 dbcAMP groups (40.1%± 5.7%, 47.5%± 6.2%, 73.1%±3.2%,and 74.3%± 6.1%)72 h after induction were significantly increased(P<0.05 or P<0.01). Compared with control group (10.2%± 2.1%), the GAD65 positive expression rates in 0.3, 0.6, 1.0,and 2.0 mmol·L-1,dbcAMP groups(22.1%±2.4%,46.9%±3.2%,70.7%±3.4%,and 72.3%±3.7%)72 h after induction were significantly increased(P<0.05 or P<0.01).Conclusion The SH-SY5Y cells have the potentiality of differentiating into GABAergic-like cells, and 1.0 mmol · L-1 is the optimal concentration of dbcAMP.

Objective To establish a method of cultivation and purification of dendritic cells(DCs) from mouse bone marrow and spleen in vitro and provide experimental material and establish basis for the study of immunological tolerance.Methods Extracted the ICR mice spleen lymphocytes and bone marrow cells under sterile conditions,and cultured DCs in co-induced of recombinant granulocyte-macrophage colony-stimulating factor(GM-CSF) and interleukin4(IL-4).Then,the changes of DC morphology were observed under light microscopy and CD80,CD86 expression levels were detected with flow cytometry.Results bone marrow cells appeared irregular and formed dendritic processes after 3 days cultured with stimulating factor in vitro which showed the typical morphology of dendritic cells,and costimulatory molecules(CD80,CD86) were low expression on the cellular suface.Conclusion Compared with the spleen cells,bone marrow cells are not only rich in DC precursor cells but also induced into DCs with a short time.

目的:探讨Nuss手术矫正漏斗胸的优越性。方法:实施漏斗胸矫形术共31例。其中10例使用胸腔镜,21例未使用胸腔镜;2例使用固定片,29例使用钢丝固定。结果:手术过程顺利,手术时间为60~90 m in,术中出血量5~15 m l,术后平均9~12 d出院,无并发症发生。出院后随诊2年,30例患者漏斗胸矫形满意,1例术后复发。矫形成功的患者中,有2例钢丝折断。结论:Nuss手术具有切口小而隐蔽、手术时间短、手术创伤小、出血少、活动早、无手术瘢痕、矫形效果好等优点,使用胸腔镜可确保术程清晰可示,手术方法安全可行,值得推广。

<正>药物临床试验是指任何在人体(患者或健康志愿者)进行药物的系统性研究,以证实或揭示试验药物的作用、不良反应及(或)试验药物的吸收、分布、代谢和排泄,目的是确定试验药物的疗效与安全性。新药需要经过Ⅰ-Ⅳ期严格的临床试验,并经过必要的审批程序后才能获批进入临床使用,药物临

Objective To analyze the effect of T2~T3 sympathicotomy in the treatment of primary palmar hyperhidrosis.Method Total 25 patients with primary palmar hyperhidrosis,including 12 males and 13 females,with the average age of 30.7 years,underwent T2~T3 sympathicotomy,in which the sympathetic chains were transected at the level of the third ribs.Results Monitoring the skin temperature of both palm.The skin temperatures of the right palm were(32.0±1.6)℃ preopratively and(34.5±1.3)℃ postoperatively(P0.05).The skin temperatures of the left palm were(32.6±1.5)℃ preopratively and(35.0±1.3)℃ postoperatively(P0.05).The procedures were successful.There were no severe intro-and post-operative complications or mortality.One case presented the Horner syndrome.All cases were followed for 6~29 months,average 16.5 months,presenting an effective rate of 100%.Conclusion Sympathicotomy is a simplified method with little side effect and higher satisfactory rate.It should be recommended for the treatment of primary palmar hyperhidrosis.

Objective To explore the effect of Heller improved operation in the treatement of achalasia of the cardia.Method Total 50 patients with achalasia of the cardia,17 males,33 females,average age of 30.7 years(2~73 years).Heller improved operation and intravenous anesthesia with tracheal intubation were used.Results For 50 cases′ operations were success and none of them appeared esophageal rupture,the recovery rate was 99.60% and without pleural effusion.Conclusion Heller improved operation is a simplified method for achalasia of the cardia with fewer side effects and higher satisfactory rate.It is effective than conventional surgery.So this is a recommended surgical method of operation.