ABSTRACT
Objective To construct a prostate cancer specific oncolytic adenovirus armed with a fusion protein gene , PSA-IZ-CD40L, and to evaluate its oncolytic efficiency and immune activation ability in vitro.Methods Prostate Specific Antigen (PSA) gene, CD40L-N and CD40L-C genes were obtained from cDNA of LNCaP cells and Jurkat cells using poly-merase chain reaction (PCR) or nested-PCR, respectively.PSA,Linker,CD40L-N and CD40L-C were linked sequentially to generate fusion protein gene PSA-IZ-CD40L (PL) by overlapping PCR.Then, prostate specific oncolytic adenovirus PL-carrying gene, Ad-PL-PPT-E1A,was constructed using the oncolytic adenovirus system , which was based on Adeasy sys-tem.PC3M cells were infected by Ad-PL-PPT-E1A at serial multiplicity of infection (MOI), and the apoptosis was detec-ted by flow cytometry at several time points post-infection.For immune activation detection , PC3M cells were infected with Ad-PL-PPT-E1A at a MOI of 50, and the cell lysate was collected at 48 h post-infection.Peripheral blood mononuclear cells derived (PBMCs) from healthy donors were stimulated by the lysate from PC 3M cells or Ad-PL-PPT-E1A infected PC3M cells before proliferation was assayed using cell counting kit-8 (CCK8).Results Fusion protein gene, PSA-IZ-CD40L, was successfully constructed and cloned into the prostate cancer specific adenovirus to generate Ad -PL-PPT-PL. The expression of E1A and PL protein could be detected by reverse transcription PCR and Western-blotting.Cytopathic effect was observed in PC3M cells infected with Ad-PL-PPT-E1A.Furthermore, the apoptosis rate reached 70.67% ± 2.98%at 48 h post-infection with 200 MOI Ad-PL-PPT-E1A.Compared with the lysate of PC3M cells, that from Ad-PL-PPT-E1A infected cells could promote the proliferation of PBMCs .Conclusion We have constructed a prostate cancer spe-cific oncolytic adenovirus armed can fusion protein gene PL , Ad-PL-PPT-E1A, which could kill PC3M cells effectively and enhance the proliferation of PBMCs in vitro.
ABSTRACT
ObjectiveTo investigate the husband and wife psychological and behavioral intervention on high-risk pregnant women,pregnancy outcome and negative emotions.Methodsin line with the number of highrisk pregnancy diagnostic criteria for pregnant women into the group of order packets were completed by the clinical observation of high-risk pregnant women in the intervention group (A) 31 cases,32 cases of high-risk pregnant women in the control group (group B),spouses of pregnant women in the intervention group (Group C) 31 spouses of pregnant women in the control group ( group D).Pregnant women in group A and group B underwent outpatient conventional high-risk pregnancy management,group A,group C received 16 weeks of husband and wife jointly participate in the key psychological problems,negative emotion coping skills to learn,couples communication skills,learning,family and social support operations,rehabilitation and faith strengthening and other intervention as the core content.Quality delivery of newborns,asphyxia,anxiety and depression in pregnant women and their spouses before and after intervention the overall incidence of anxiety and self-assessment scale(SAS) score,the score of the Self-Rating Depression Scale (SDS),the Family APGAR Index Questionnaire score (observation of high-risk pregnancy APGAR) and other changes.ResultsThe average body weight of newborns:the intervention group A (3.12 ± 0.69) kg,than in group B (2.29 ± 0.78) kg,a statistically significant difference ( t =2.3148,P =0.024) ;asphyxia:group A was 12.9％ and 34.4％ in group B,the difference was statistically significant (x2 =4.0018,P=0.0455) ;natural birth rate:58.1％ in group A,group B 25％,a statistically significant difference (x2 =7.1023,P=0.0077) ;the rate of cesarean section:29.0％ in group A,group B,59.4％,a statistically significant difference ( x2 =5.8713,P =0.0154 ) ; anxiety and depression in pregnant women:the total incidence after the intervention group A was 19.4％,46.9％ in Group B,the difference was significant (x2 =5.3664,P=0.0205) ;maternal spouse anxiety and depression:in the overall incidence of A group of 9.7％ after the intervention group B 31.2％,the difference was statistically significant (x2 =4.4745,P =0.0344 ) ;APGAR score:after the intervention of high-risk pregnant women in group A (9.42 ± 1.53),Group B (7.71 ± 1.56),group A better than group B,the difference was statistically significant ( t =4.3910,P =0.000),intimacy,emotional degree,the growth degree,cooperation degree,adapt to the degree of five factor scores in group A than group B,a statistically significant difference (P ＜ 0.05,P ＜ 0.01 ).ConclusionHigh-risk pregnant women and their spouses have a severe negative emotional reaction,the husband and wife psychological and behavioral intervention on the improvement of high-risk pregnant women,pregnancy outcome and negative emotions have an important role.
ABSTRACT
Objective: To establish the quality standard for Huahaiyigan Effervescent Granules. Methods: Astragalus membranaceus(Fisch) Beg., Bupleurum chinenes DC., Polygonum cuspidatum Sieb. et Zucc. and Schisandra chinensis(Turcz.) Baill. were identified by TLC; and the content of schisandrin B was determined by HPLC. Results: Astragalus membranaceus(Fisch.) Beg., Bupleurum chinenes DC., Polygonum cuspidatum Sieb. et Zucc. and Schisandra chinensis(Turcz.) Baill. could be identified by TLC; schisandrin B showed a good linear relationship at a range of 0.848～4.240?g, r=0.9999; The average recovery was 98.16% and RSD was 1.12%. Conclusion: This method was available for quality control of Huahaiyigan Effervescent Granules