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PURPOSE:To detect the protein expression level of foreign fused gene in the first generation of genetically modified tomatoes transformed with fused gene of antigen epitope PacP in surface protein of Streptococcus mutans and cholera toxin B subunit. METHODS: The total DNA in tomatoes was extracted and PCR was applied to screen the first generation of transgenic tomatoes carrying Streptococcus mutans surface protein coding PAcP with cholera toxin B subunit fusion gene. Total proteins were extracted and quantitatively tested with BAC kit. Foreign fused protein expression level was analysed by Western blotting and ELISA. RESULTS: Ten DNA samples with the full length of 1.6 kb foreign fused DNA fragment was detected by PCR among all 18 samples. The crude protein in the transgenic tomato fruit tissue was 3.93 mg/mL. Compared with the normal tomatoes, Western blotting showed that the transgenic tomato protein contained a distinct protein band with a molecular weight about 60 kD. The results of ELISA suggested that fused foreign protein level was up to 0.18% of the total soluble protein in genetically modified tomato fruit. CONCLUSIONS: The target protein encoded by fused foreign gene of antigen epitope PacP in surface protein and cholera toxin B subunit was expressed in genetically modified tomatoes fruit. Supported by National Natural Science Foundation of China (30160086), Science and Technology Support Project of Guizhou Province [SY (2012)3086], The Sixth Batch of Technology Innovation Team Project in Guizhou Province[(2013) 4026] and Science and Technology Foundation of Guizhou Province [LKZ(2011)41]. [ABSTRACT FROM AUTHOR]

PURPOSE: To investigate the efficiency of pcDNA3-PAc on Japanese long-eared white rabbits by intranasal immunization, and observe the appreciate gene vaccine dose in rabbit immunization. METHODS: Thirty Japanese long-eared white rabbits were randomly divided into 5 groups (6 in each group) as follows: 200, 400, 600 µg pcDNA3-PAc plasmid group; 400 µg pcDNA3 group and inactivated whole-cell vaccine positive control group. The rabbits were immunized twice, and plasmid groups and whole-cell group were coupled Freund's adjuvant with 1:1 ratio. The specific IgG and S-IgA antibodies in serum and saliva were detected with indirect ELISA method. The data was analyzed with SPSS 17.0 software package. RESULTS: ➀ The peak time of the antibodies appeared between 8-10 weeks after the first immunization; ➁ The specific antibodies IgG and S-IgA could be detected 2 weeks after immunization; ➂ The level of salivary specific S-IgA and serum specific IgG of pcDNA3-PAc were significantly higher than negative groups (P<0.05). The titer of the 200 µg was significantly lower than those of 400 µg and 600 µg group (P<0.05). CONCLUSIONS: ➀ The recombinant plasmid pcDNA3-PAc has immunogenicity, which can induce specific immune responses for 14 weeks in rabbits. ➁ The results of the present study show that 200 µg, 400 µg and 600 µg are effective immunizational dosage to 1.5 kg Japanese long-eared white rabbits. ➂ 400 µg and 600 µg pcDNA3-PAc can be considered as the optimal dosage than 200 µg at present experimental condition. [ABSTRACT FROM AUTHOR]

PURPOSE: To detect the exogenous gene copy number of the transgenic tomato anti-caries vaccine by using the SYBR Green real-time PCR. METHODS: Recombinant plasmid pEACIO and pEPCIO were used as standard to detect genome samples of exogenous gene pacA-ctxB and pacP-ctxB by SYBR green fluorescent quantitation, then the average value was calculated as gene copy number. RESULTS: The copy number of the transgenic tomato carrying pacA-ctxB was 1.3 and the pacP-ctxB was 3.2. CONCLUSIONS: The transgenic tomato plants which have high stability are low-copy transgenic plants. Supported by National Natural Science Foundation of China (30160086, 81260164), Science and Technical Fund of Guizhou Province (LKZ[2011]41), Project of Technology Innovation Team in Guizhou Province, Leading Academic Discipline Construction Project in Guizhou Province and Excellent Scientific Research Team Cultivation Project in Zunyi Medical College ([2012] 12). [ABSTRACT FROM AUTHOR]

Background: Portal shunt and immune status related to the spleen are related to the occurrence of hepatic encephalopathy (HE). It is unknown whether spleen volume before transjugular intrahepatic portosystemic shunt (TIPS) is related to postoperative HE., Aim: To investigate the relationship between spleen volume and the occurrence of HE., Methods: This study included 135 patients with liver cirrhosis who underwent TIPS, and liver and spleen volumes were elevated upon computed tomography imaging. The Kaplan-Meier curve was used to compare the difference in the incidence rate of HE among patients with different spleen volumes. Univariate and multivariate Cox regression analyses were performed to identify the factors affecting overt HE (OHE). Restricted cubic spline was used to examine the shapes of the dose-response association between spleen volumes and OHE risk., Results: The results showed that 37 (27.2%) of 135 patients experienced OHE during a 1-year follow-up period. Compared with preoperative spleen volume (901.30 ± 471.90 cm 3 ), there was a significant decrease in spleen volume after TIPS (697.60 ± 281.0 cm 3 ) in OHE patients. As the severity of OHE increased, the spleen volume significantly decreased ( P < 0.05). Compared with patients with a spleen volume ≥ 782.4 cm 3 , those with a spleen volume < 782.4 cm 3 had a higher incidence of HE ( P < 0.05). Cox regression analysis showed that spleen volume was an independent risk factor for post-TIPS OHE (hazard ratio = 0.494, P < 0.05). Restricted cubic spline model showed that with an increasing spleen volume, OHE risk showed an initial increase and then decrease ( P < 0.05)., Conclusion: Spleen volume is related to the occurrence of OHE after TIPS. Preoperative spleen volume is an independent risk factor for post-TIPS OHE., Competing Interests: Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

Purpose: To detect the protein expression level of foreign fused gene in the first generation of genetically modified tomatoes transformed with fused gene of antigen epitope PacP in surface protein of Streptococcus mutans and cholera toxin B subunit., Methods: The total DNA in tomatoes was extracted and PCR was applied to screen the first generation of transgenic tomatoes carrying Streptococcus mutans surface protein coding PAcP with cholera toxin B subunit fusion gene. Total proteins were extracted and quantitatively tested with BAC kit. Foreign fused protein expression level was analysed by Western blotting and ELISA., Results: Ten DNA samples with the full length of 1.6 kb foreign fused DNA fragment was detected by PCR among all 18 samples. The crude protein in the transgenic tomato fruit tissue was 3.93 mg/mL. Compared with the normal tomatoes, Western blotting showed that the transgenic tomato protein contained a distinct protein band with a molecular weight about 60 kD. The results of ELISA suggested that fused foreign protein level was up to 0.18% of the total soluble protein in genetically modified tomato fruit., Conclusions: The target protein encoded by fused foreign gene of antigen epitope PacP in surface protein and cholera toxin B subunit was expressed in genetically modified tomatoes fruit.

Purpose: To investigate the efficiency of pcDNA3-PAc on Japanese long-eared white rabbits by intranasal immunization, and observe the appreciate gene vaccine dose in rabbit immunization., Methods: Thirty Japanese long-eared white rabbits were randomly divided into 5 groups (6 in each group) as follows: 200, 400, 600 μg pcDNA3-PAc plasmid group; 400 μg pcDNA3 group and inactivated whole-cell vaccine positive control group. The rabbits were immunized twice, and plasmid groups and whole-cell group were coupled Freund's adjuvant with 1:1 ratio. The specific IgG and S-IgA antibodies in serum and saliva were detected with indirect ELISA method. The data was analyzed with SPSS 17.0 software package., Results: (1) The peak time of the antibodies appeared between 8-10 weeks after the first immunization; (2)The specific antibodies IgG and S-IgA could be detected 2 weeks after immunization;(3)The level of salivary specific S-IgA and serum specific IgG of pcDNA3-PAc were significantly higher than negative groups (P<0.05). The titer of the 200 μg was significantly lower than those of 400 μg and 600 μg group (P<0.05)., Conclusions: (1)The recombinant plasmid pcDNA3-PAc has immunogenicity, which can induce specific immune responses for 14 weeks in rabbits. (2)The results of the present study show that 200 μg, 400 μg and 600 μg are effective immunizational dosage to 1.5 kg Japanese long-eared white rabbits. (3)400 μg and 600 μg pcDNA3-PAc can be considered as the optimal dosage than 200 μg at present experimental condition. Supported by Guizhou Science and Technology Projet of Distinguished Young Talents [2005(0509)], Guizhou College and University Leading Academic Discipline Project [2012(15)] and Zunyi Medical University Project of Distinguished Research Team [2012(12)].

Purpose: To detect the exogenous gene copy number of the transgenic tomato anti-caries vaccine by using the SYBR Green real-time PCR., Methods: Recombinant plasmid pEAC10 and pEPC10 were used as standard to detect genome samples of exogenous gene pacA-ctxB and pacP-ctxB by SYBR green fluorescent quantitation, then the average value was calculated as gene copy number., Results: The copy number of the transgenic tomato carrying pacA-ctxB was 1.3 and the pacP-ctxB was 3.2., Conclusions: The transgenic tomato plants which have high stability are low-copy transgenic plants. Supported by National Natural Science Foundation of China (30160086, 81260164), Science and Technical Fund of Guizhou Province (LKZ[2011]41), Project of Technology Innovation Team in Guizhou Province, Leading Academic Discipline Construction Project in Guizhou Province and Excellent Scientific Research Team Cultivation Project in Zunyi Medical College ([2012]12).