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OBJECTIVE:To study the effects of rAd-p53 injection on the proliferation,apoptosis and autophagy of nephroblas-toma cells. METHODS:Nephroblastoma cells were respectively cultured 20 h with high,medium,low concentration(1×109,1× 108,1×107 VP/mL)of Recombinant human rAd-p53 injection,cells with no injection were the blank control. Cell proliferation,cy-cle,apoptosis and related gene p21,Bax protein expressions were detected,and autophagy gene LC-3,Atg7,Atg12 expressions and number of autophagosomes were also detected. RESULTS:The proliferation inhibition rates of high,medium,low concentra-tion of Recombinant human rAd-p53 injection to nephroblastoma cells were(42.86±3.18)%,(33.64±7.25)%,(16.26±9.07)%;apoptotic rates were (53.85 ± 9.36)%,(37.35 ± 9.64)%,(23.64 ± 10.65)%,respectively. Compared with blank control,cells at period G0/G1 were increased under high,medium,low concentration Recombinant human rAd-p53 injection,effects were relatively obvious under high,medium concentration (P0.05). CONCLUSIONS:Recom-binant human rAd-p53 injection can inhibit the cell proliferation of nephroblastoma,and induce its apoptosis and autophagy.
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BACKGROUND:Retinoic acid is the most promising inducer for neuroblastoma minimal residual lesion, and it can induce cel differentiationin vivo, accompanied by reducing tumor cel proliferation. OBJECTIVE:To study the effect of nanoparticle labeling on biological characteristics of neuroblastoma stem cels, and the role of 13-cis retinoic acid to induce differentiation of neuroblastoma stem cels. METHODS:Neuroblastoma stem cels were isolated and culturedin vitro using serum-free suspension culture method, labeled with polylysine-modified γ-Fe2O3 nanoparticles and induced in culture medium containing 13-cis retinoic acid. RT-PCR was used to detect the expression of Oct-4 before and after labeling as wel as before and after induction. Immunofluorescence method was used to detect the expression of nestin before and after labeling as wel as before and after induction. RESULTS AND CONCLUSION:Neuroblastoma stem cels were successfuly cultured in the bone marrow samples from 5 of 20 cases. Polylysine-modified γ-Fe2O3 nanoparticle labeling did no influence the viability and proliferation ability of neuroblastoma stem cels, and also had no effect on Oct-4 mRNA and nestin expression. After cultured in the culture medium containing 13-cis retinoic acid, the cel shape changed and the growth rate slowed down. Moreover, the expression of Oct-4 mRNA and nestin was gradualy reduced. These findings indicate that polylysine-modified gamma-Fe2O3 nanoparticles can be used to label neuroblastoma stem cels, and 13-cis retinoic acid can induce the differentiation of neuroblastoma stem cels.
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Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs.
Subject(s)
Animals , Female , Mice , Antibodies, Viral , Blood , CD4-Positive T-Lymphocytes , Cell Biology , Allergy and Immunology , CD8-Positive T-Lymphocytes , Cell Biology , Allergy and Immunology , Cell Proliferation , Classical Swine Fever Virus , Genetics , HSP70 Heat-Shock Proteins , Genetics , Allergy and Immunology , Haemophilus parasuis , Genetics , Immunization , Interferon-gamma , Metabolism , Mice, Inbred BALB C , Plasmids , Genetics , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Viral Envelope Proteins , GeneticsABSTRACT
To study the chemical constituents of Senecio nemorensis.,12 known compounds were isolated and identified,which were niacinamide(Ⅰ),vanillin(Ⅱ),syringic acid(Ⅲ),syringalddehyde(Ⅳ),3-acetyl-4-hydroxy-benzoic acid(Ⅴ),4,4-dimethyl-1,7-heptanedioic acid(Ⅵ),1H-indole-3-carboxaldehyde(Ⅶ),ethyl caffeate(Ⅷ),1-0-(E)-p-methoxycinnamoyl-β-D-glucopyranoside(Ⅸ),(6S,7E)-6-hydroxy-4,7-megastigmadien-3,9-dione(Ⅹ),Annuionone D(Ⅺ),and(1 'S,6'R)-abscisic acid(Ⅻ),respectively.
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Objective To study the anatomy of splenic hilum blood vessels in order to thread ligature(endoligature)instead of using stapler during the process of laparoscopic splenectomy and to evaluate the prelimnary clinical results.Methods 41 children patients underwent laparoscopic splenectomy with this technique(endoligature)for various hematologic and autoimmune disorders,including 25 cases of hereditary spherocytosis,13 idiopathic thrombocytopenia purpura,and 3 hypersplenic granulocytopenia.The anatomy of splenic pedicle,the adjacent relation between splenic vessel and pancreas were detected by color Doppler ultrasonography preoperatively.The above-mentioned parameters were compared with that found intraoperatively.Results The relationships of splenic vessel and pancreas was of type Ⅰ in 24 cases and type Ⅱ in 17.In 31 cases,the major splenic blood vessels were ramified into branches 2 cm away from the hilum and in 10 it was within 2 cm as detected by preoperative ultrasonography.These characters were largerly identified by laparoscopic laparotomy,and in all the 41 cases laparoscopic splenectomy was successfully accomplished using this endoligature instead of vasculature stapler.There was no serious complication.The mean operating time was(114 ±31)min,the estimated blood loss was(51 ±23)ml.Conclusions Ultrasonography could identify the anatomic type of splenic vessel,and its relation with the pancreas.Endoligature in the management of splenic pedicles during laparoscopic splenectomy is safe and reliable.
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Objective To explore the feasibility and t he efficacy of laparoscopic splenectomy of massive splenomegaly in the treatment o f hereditary spherocytosis in children. Methods Seven children with massive splenomegaly underwent laparoscopic splenectomy. Their age ranged 1~14 years (mean, 8.8 years), and their body weight was 10~57 kg (mean, 33.8 kg) . Three children had an accompanying cholelithiasis: 2 of them received a concom itant cholecystectomy and 1 of them, cholecystotomy. Results A ll the operations were successfully performed under laparoscope, and hand-assist ed splenectomy through a small incision was applied in 1 child because of bleedi ng of the splenic vein. The duration of operation was 50~150 min (mean, 90 min), the intraoperative blood loss was 30~500 ml (mean, 117 ml), and the length of h ospitalization, 4~10 d (mean, 5.5 d). The red blood cell counts had significantl y increased 3 days following the operation (t=2.652, P
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Objective To evaluate intraoperative cholangiopancreatography in understanding pathologic changes of congenital choledochal cyst(CCC) with anomalous junction of pancreaticobiliary duct(APBD) and for rational operative procedure. Methods Eighty-two CCC cases were examined by intraoperative cholangiopancreatography(ICP). The morphologies of the biliary tract and types of APBD were recorded. Results Of the 82 CCC cases there were 35 cases of Todani Ia,9 cases of Ib,28 cases of Ic and 10 cases of Ⅳ.APBD was diagnosed in 73 cases and classified according to Komi classification. Of them,37 cases were classified as type I of APBD (bile duct drains into pancreatic duct),30 cases as type II (pancreatic duct drains into bile duct) and 6 cases as type III (complicated APBD with patent accessory pancreatic duct). Of the 44 cases with choledochal cystic dilatations,33 cases were of type I. Of the 29 biliary fusiform dilatations,21 were of type II. Of 21 patients with pancreatitis,17 were of type II (? 2=33.14,P