To investigate the technique of sorting high-purity precartilaginous stem cells from rat's perichondrium, neonatal rat's perichondrium cells suspensions were incubated with monoclone antibody of anti-fibroblast growth factor receptor-3 (anti-FGFR-3), and the labeled cells were separated from the suspension in the magnetic field by immuno-beads coated with the second antibody. Purity of the sorted neural stem cells was found to be 93.0%-99.0%, with living cells amounting to 80% -85 %. The magnetic cell sorting system could effectively separate precartilaginous stem cells from perichondrium cell suspensions.

Using gene fusion technology, polypeptide fusion tags can be engineered into target proteins at the genetic level.The resultant recombinant proteins may possess the biochemical properties of the imported fusion tags.Therefore, it is possible to take advantage of fusion tags to improve and evaluate protein expression, to detect and track protein targets, and to purify and characterize proteins.However, it is necessary to eliminate any influence of the fusion tag in structural characterization experiments or in isolating pharmaceutical proteins.Scientists must therefore remove fusion tags prior to structural and functional analyses when fusion tags are suspected of interfering with the biological activity of a protein or influencing its behavior.The fusion tag can be removed by several methods including harsh chemical treatment, mild enzymatic cleavage by endoprotease or exoprotease, and intein-mediated self-cleavage.Here the literature is reviewed in relation to principles, applications, and approaches of each method.

Objective To investigate the clinical features,diagnosis,treatment and prognosis of gastric neuroendocrine carcinoma (G-NEC).Methods Clinical data of 52 G-NEC cases were analyzed.Follow-up was conducted by telephone.The survival curves were drawn using Kaplan-Meier method.Univariate analysis was performed by the Log-rank test and multivariate analysis was performed by COX proportional hazards model.Results The median overall survival rate was 19 months (range 6 to61 months),and the overall 1,3,5-year survival rates were 74％,16％ and 5％ respectively.Tumor stage surgery,vascular nerve involvement and Ⅲ and Ⅳ phase chemotherapy were related to prognosis (x2 =24.254,10.005,7.261,8.790,all P ＜ 0.05).Multivariate analysis showed tumor stage and vascular nerve involvement were independent prognostic factors (x2 =17.170,5.810,all P ＜ 0.05).Conclusion G-NEC is a highly malignant tumor with poor prognosis.Preoperative diagnosis rate is low.Surgery is the treatment of first choice.Definite diagnosis depends on postoperative pathology and immunohistochemical examination.

Herpes simplex virus (HSV) is a double-stranded DNA virus that can infect skin and mucosal epithelial cells. It can establish latency in sensory neurons and sporadically reactivate from these cells. In order to reply to attacks of the host and evade the immunity surveillance during infection and reactivation, HSV has developed a multitude of clever strategies. Dendritic cells (DCs), one of the most important antigen-presenting cells (APC), can recognize pathogens at the infection sites and activate specific T cells, thus playing a crucial role in the host immunity against virus infection. This paper reviewed the mechanism of the host immunity against HSV, especially the role of DCs in HSV-induced immune responses and the future research perspective.

95%). Conclusion Immunomagnetic separation can effectively isolate and purify PSCs.

BACKGROUND:There is a lack of study on material properties and parameters of foot finite element models in China. Vernier caliper is a common method for measuring the width and thickness of ligaments and tendons to calculate the cross-sectional area.
OBJECTIVE:To design a new ligament cross-sectional area measuring instrument to improve the measurement accuracy.
METHODS:The cross-sectional area of the five fresh cadaver ankle ligaments was respectively measured using the new instrument and vernier caliper, and then a comparative analysis of the two measurement methods was performend.
RESULTS AND CONCLUSION:The cross-sectional area of anterior talofibular ligament, calcaneofibular ligament, tibionavicular ligament and calcaneotibial ligament was (20.61±7.52), (22.38±11. 49), (33.09±9.91) and (28.20±10.88) mm2, respectively measured by the vernier caliper, and (17.59±4.03), (20.77±7.91), (28.08±8.14) and (30.39±7.98) mm2 by the new ligament cross-sectional area measuring instrument. These results suggest that this new measuring instrument is accurate, reliable and easy to operate, which can be used as a special instrument to measure ligament cross-sectional area, but further studies wil be necessary.

Immortalized human precartilaginous stem cells (IPSCs) were established to provide stable cell resource for the study of the molecular mechanism of gene targeting on the differentiation of PSCs. Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs by using lipofectin transfection. Colonies were isolated by puromycin selection and expanded by multiple passages. Immunohistochemistry, RT-PCR and Southern blotting were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines. The positive colonies were isolated and subcultured, designated immortalized precartilaginous stem cells (IPSCs), which were confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive cells by immunohistochemistry and RT-PCR. SV40Tag cDNA was found in cultured IPSCs of passage 8 by Southern blotting, and the expressions of SV40Tag mRNA and protein were confirmed by RT-PCR. These findings suggested that IPSCs strain with SV40Tag was constructed successfully.
Cartilage/*cytology ; Cell Proliferation ; Cell Transformation, Viral ; Cells, Cultured ; Fetus ; Simian virus 40/*genetics ; Stem Cells/*cytology ; Transfection

OBJECTIVETo explore the clinical effects of negative pressure wound therapy (NPWT) combined with artificial dermis grafting and autologous skin grafting on repair of open joint wounds and/or wounds with exposed bone fracture.

METHODSEleven patients with open joint wounds and/or wounds with exposed bone fracture, hospitalized from November 2008 to November 2014, were enrolled in the study. According to the differences of the first stage treatment, all patients were divided into experimental group ( n = 6, including 4 patients of open joint wounds, 1 patient of wound with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture), and control group ( n 5, including 2 patients of open joint wounds, 2 patients of wounds with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture). After debridement, the wounds in both groups were grafted with punctured artificial dermis, while NPWT was only used over the artificial dermis of experiment group for 1 week. In the operation at sacsod stage, autologous split-thickness skin was grafted on the vascularized artificial dermis in both groups. Results In 5 patients of open joint wounds in experimental group, the artificial dermis was vascularized well, autologous skin grafts survived, and wounds were healed. In 3 patients of open joint wounds in control group, the artificial dermis grafting all failed due to local infection, and then these wounds were repaired with local tissue flap grafting. Artificial dermis in 3 patients of wounds with exposed bone fracture in both groups was vascularized well after grafting, and the wounds were healed after autologous skin grafting, whether or not NPWT was used.

CONCLUSIONSNPWT combined with artificial dermis grafting and autolognus skin grafting can be used for repairing open joint wounds and/or wounds with exposed bone fracture.

Debridement ; Dermis ; transplantation ; Humans ; Negative-Pressure Wound Therapy ; methods ; Skin Transplantation ; methods ; Skin, Artificial ; Surgical Flaps ; Wound Healing

Equipment Failure ; Female ; Heart Ventricles ; pathology ; surgery ; Humans ; Middle Aged ; Pacemaker, Artificial

Objective To clone and express bradyzoite antigen 1(BAG1) gene of T. gondii,and analyze the immunoreactivity of the recombinant product. Methods The differentiation of T. gondii RH strain tachyzoites into bradyzoites was induced in vitro,and the coding sequence of BAG1 was amplified from bradyzoites by RT-PCR. The PCR product was analyzed by sequencing. The BAG1 coding sequence was further subcloned into the plasmid pET32a(+). The plasmid pET32a(+)-BAG1 was then transformed into BL21(DE3) to express after IPTG induction. The expression product was purified with Ni-NTA agarose and the purified BAG1 was further analyzed by Western blotting and ELISA. Results BAG1 cDNA was amplified from bradyzoites. After IPTG induction,BAG1 was expressed in a fusional form in E. coli. Western blotting showed that the purified recombinant protein could be specifically recognized by sera from mice chronically infected by T. gondii B36 strain. ELISA showed that the positive rate of T. gondii IgG antibodies of 350 human sera detected by the recombinant BAG1(17.4%) was higher than by recombinant SAG1 (12.6%)(P