BACKGROUND: It is one of hot topics for the application of neurotrophic factors including neurenergen-3 to promote peripheral neural regeneration nowadays; however, clinical application is restricted to safety and effective administration route. Gene transfection brings a novel thinking and pathway for neurotrophic factors used in clinic.OBJECTIVE: To observe the expression of neural stem cells modified by neurenergen-3 gene after transfection.DESIGN: Completely randomized study.SETTING: Department of Traumatic Orthopaedics and Hand Surgery, the First Affiliated Hospital, Guangxi Medical University;Department of Hand Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Three healthy SD rats of four months old and either gender were selected from Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Recombinant adenoviral expressing vector for transfection of neural stem cells was constructed in Laboratory of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, and the concentration was 0.15 g/L.METHODS: The experiment was carried out in the Orthopaedic Laboratory and Central Laboratory, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from December 2002 to March 2004. Recombinant plasmid pAD-neurenergen-3 containing with green fluorescent protein (GFP) gene was transfectd into primarily cultured neural stem cells by using cationic tiposome interventional method. At 72 hours after transfection, fluorescent inverted microscope was used to observe GFP expression in neural stem cells, and transfection efficiency was measured simultaneously. Expression and transcription of neurenergen-3 gene in neural stem cells were detected at 72 hours, 1 and 5 weeks after transfection by using immunocytochemical stain and reverse transcription polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of neurenergen-3 gene in transfected neural stem cells.RESULTS: ① Neural stem cells transfected by recombinant plasmid pAD-neurenergen-3: GFP expressed on partial neural stem cells at 72 hours after transfection, and the transfection efficiency was 40%. Five weeks later, GFP expression was still observed. ②Transcription and expression of neurenergen-3 gene in neural stem cells: Transcription of neurenergen-3 mRNA was observed in neural stem cells at 72 hours, 1 and 5 weeks after transfection, and expression of neurenergen-3 mRNA was still observed in 5 weeks after trans fection.CONCLUSION: As the carrier of cationic liposome, neurenergen-3 gene can effectively transfect, culture and long-term express neural stem cells via the introduction of adenoviruses.

Objective To study the feasibility and safety of measles vaccination in women of childbearing age and to understand the influence of measles vaccination on the fetal transmission measles antibody level of the infants.Methods From January 1,2012 to December 31,2012,600 women of childbearing age were included in this study.The measles IgG antibody was detected,then all participants were randomized according to the IgG level.The measles IgG antibody of participants in non-vaccinated group was detected predelivery in hospital by enzyme linked-immunosorbent assay (ELISA).That of participants in vaccinated group was detected 3 months after vaccination with measles mumps rubella combined vaccine and predelivery by ELISA.And measles nuclear protein fragment gene (measles virus nucleoprotein,MVN) in the blood was detected by reverse transcription polymerase chain reaction (RT PCR).A total of 368 participants gave birth within 2 years after vaccination,and 357 infants 8 months after birth were healthy.In non-vaccinated group,8 infants were diagnosed with measles within 8 months after birth.Finally,349 8-month infants were enrolled in the study,including 52 whose mothers in high antibody without vaccination group,65 whose mothers in high antibody with vaccination group,110 whose mothers in low antibody without vaccination group and 122 whose mothers in low antibody with vaccination group.The measles IgG antibody levels in the blood of all the 8-month infants were detected.Data were analyzed using t test,one-way ANOVA (Newman-Keuls was used for comparison between groups) and Pearson analysis.Results The measles IgG antibody level of expectant women in high antibody without vaccination group was significantly lower compared to that in high antibody with vaccination group ([268.5±74.9] IU/mL vs [578.3t208.1] IU/mL,Q=15.57,P＜0.01).That in low antibody without vaccination group was also significantly lower than low antibody with vaccination group ([169.4+42.3] IU/mL vs [584.7+195.8] IU/mL,Q=29.54,P＜0.01).The results of MVN RT-PCR after 3 months of vaccination showed no positive bands in all blood samples.Two of the expectant women in high antibody without vaccination group and one in low antibody without vaccination group were positive for MVN bands.Among 8-month infants,the levels of antibody in high antibody without vaccination group and high antibody with vaccination group were (106.3 ± 36.8) IU/mL and (291.8±86.5) IU/mL,respectively,which was statistically different (t=23.33,P＜0.01).Those in low antibody without vaccination group and low antibody with vaccination group were (87.1 ± 26.4) IU/mL and (274.0±72.5) IU/mL,respectively,which was statistically different (t =33.27,P＜0.01).The measles antibody level of expectant women was positively correlated with their 8-month infants (r=0.652,P＜0.01).All 8 infants who were diagnosed with measles were delivered by women without vaccination,and the measles infection rate of infants was significantly different between women with and without vaccination (P=0.002).Condusion It is feasible for women of childbearing age to receive measles vaccination,which can increase the measles IgG antibody level of both expectant women and their infants.