1.Preparation of monoclonal antibody against bovine viral diarrhea virus and estab-lishment of double antibody sandwich ELISA method
Qianyue MA ; Jiaxuan LI ; Yanping JIANG ; Wen CUI ; Xinyuan QIAO ; Changcheng ZHU ; Shize HAO
Chinese Journal of Veterinary Science 2025;45(11):2343-2350
The purpose of this study was to prepare high affinity monoclonal antibodies(mAbs)a-gainst bovine viral diarrhea virus(BVDV)and establish a double antibody sandwich ELISA detec-tion method.BVDV was purified by differential ultracentrifugation and used to immunize BALB/c mice.Hybridoma cells were prepared by fusing spleen cells from the immunized mice with SP2/0 cells.Positive cells were screened by indirect ELISA.A double-antibody sandwich ELISA method for detecting BVDV was developed using monoclonal antibody 4D11 as the capture antibody and HRP-labeled monoclonal antibody 3F3 as the detection antibody.The results of the ELISA and the determination of the variable region gene sequence of monoclonal antibodies indicated that the two monoclonal antibodies recognize different antigenic epitopes.Specificity tests showed that two monoclonal antibodies specifically recognize BVDV and did not cross-react with other bovine viru-ses associated with diarrhea.Indirect immunofluorescence assay and Western blot assay demonstra-ted that both mAbs exhibited strong reactivity with BVDV.The double antibody sandwich ELISA detection method established in this study had good specificity.The sensitivity test revealed that the method could detect a minimum virus amount of 3.1 × 104 TCID50.The reproducibility test showed that the inter-batch coefficient of variation(Cv)was between 2.47%and 7.44%,and the intra-batch Cv was between 1.71%and 9.89%,indicating good reproducibility.The establishment of this method provides an effective technical tool for the rapid diagnosis and prevention and con-trol of BVDV.
2.Preparation of monoclonal antibody against bovine viral diarrhea virus and estab-lishment of double antibody sandwich ELISA method
Qianyue MA ; Jiaxuan LI ; Yanping JIANG ; Wen CUI ; Xinyuan QIAO ; Changcheng ZHU ; Shize HAO
Chinese Journal of Veterinary Science 2025;45(11):2343-2350
The purpose of this study was to prepare high affinity monoclonal antibodies(mAbs)a-gainst bovine viral diarrhea virus(BVDV)and establish a double antibody sandwich ELISA detec-tion method.BVDV was purified by differential ultracentrifugation and used to immunize BALB/c mice.Hybridoma cells were prepared by fusing spleen cells from the immunized mice with SP2/0 cells.Positive cells were screened by indirect ELISA.A double-antibody sandwich ELISA method for detecting BVDV was developed using monoclonal antibody 4D11 as the capture antibody and HRP-labeled monoclonal antibody 3F3 as the detection antibody.The results of the ELISA and the determination of the variable region gene sequence of monoclonal antibodies indicated that the two monoclonal antibodies recognize different antigenic epitopes.Specificity tests showed that two monoclonal antibodies specifically recognize BVDV and did not cross-react with other bovine viru-ses associated with diarrhea.Indirect immunofluorescence assay and Western blot assay demonstra-ted that both mAbs exhibited strong reactivity with BVDV.The double antibody sandwich ELISA detection method established in this study had good specificity.The sensitivity test revealed that the method could detect a minimum virus amount of 3.1 × 104 TCID50.The reproducibility test showed that the inter-batch coefficient of variation(Cv)was between 2.47%and 7.44%,and the intra-batch Cv was between 1.71%and 9.89%,indicating good reproducibility.The establishment of this method provides an effective technical tool for the rapid diagnosis and prevention and con-trol of BVDV.
3.Efficacy and safety of oral atenolol for the treatment of infantile haemangiomas: a single-arm meta-analysis
Zhisheng ZHU ; Xianying ZHOU ; Xingong LIN ; Shiquan WU ; Chaoyang WANG ; Shize ZHU
Chinese Journal of Plastic Surgery 2023;39(11):1192-1203
Objective:To investigate the efficacy, safety and recurrence rate of oral atenolol in the treatment of infantile hemangioma, so as to provide evidence-based medicine basis and reference for clinic.Methods:Search on the following public databases from January 1, 2008 to June 13, 2022: Web of Science, PubMed, Cochrane Library, Embase, U. S. National Library of Medicine Clinical Trials Registry Platform; China National Knowledge Infrastructure(CNKI), Chinese Biomedical Literature Service System(SinoMed), Chinese Science and Technology Journal Database and Wanfang Data. According to inclusion and exclusion criteria, studies on oral atenolol for the treatment of infantile hemangioma were selected. The outcome indicators were efficiency (complete response rate), incidence of adverse effects and recurrence rate. The single-arm meta-analysis was performed using R software version 4.1.2. Egger’s test was employed and funnel plots were drawn to assess publication bias in the literature.Results:A total of 14 studies were included, comprising 5 randomized controlled trials, 5 single-arm studies, 3 non-randomized controlled trials, and 1 case-control study. The oral administration of atenolol for the treatment of infantile hemangiomas resulted in an efficacy rate (complete remission rate) of 62% (95% CI 52%~71%). The incidence rate of adverse reactions related to the digestive system was 18% (95% CI 7%~30%), while that related to β2 receptor blockade was 4% (95% CI 2%~6%), central nervous system-related adverse reactions occurred at a rate of 10% (95% CI 5%~16%), the recurrence rate was 5% (95% CI 2%~9%). Egger’s test indicated that there was no significant publication bias in the efficacy rate, central nervous system-related adverse reaction rate, and gastrointestinal-related adverse reaction rate of oral atenolol treatment for infantile hemangiomas ( P>0.05). The sensitivity analysis for the efficacy rate, adverse reaction rate, and recurrence rate of oral atenolol treatment for infantile hemangiomas suggested that the result were stable and reliable. Conclusion:Oral administration of atenolol for the treatment of infantile hemangiomas demonstrates significant efficacy, fewer adverse reactions, and a low recurrence rate, making it a promising candidate as a reasonable alternative to oral propranolol for treating infantile hemangiomas.
4.Efficacy and safety of oral atenolol for the treatment of infantile haemangiomas: a single-arm meta-analysis
Zhisheng ZHU ; Xianying ZHOU ; Xingong LIN ; Shiquan WU ; Chaoyang WANG ; Shize ZHU
Chinese Journal of Plastic Surgery 2023;39(11):1192-1203
Objective:To investigate the efficacy, safety and recurrence rate of oral atenolol in the treatment of infantile hemangioma, so as to provide evidence-based medicine basis and reference for clinic.Methods:Search on the following public databases from January 1, 2008 to June 13, 2022: Web of Science, PubMed, Cochrane Library, Embase, U. S. National Library of Medicine Clinical Trials Registry Platform; China National Knowledge Infrastructure(CNKI), Chinese Biomedical Literature Service System(SinoMed), Chinese Science and Technology Journal Database and Wanfang Data. According to inclusion and exclusion criteria, studies on oral atenolol for the treatment of infantile hemangioma were selected. The outcome indicators were efficiency (complete response rate), incidence of adverse effects and recurrence rate. The single-arm meta-analysis was performed using R software version 4.1.2. Egger’s test was employed and funnel plots were drawn to assess publication bias in the literature.Results:A total of 14 studies were included, comprising 5 randomized controlled trials, 5 single-arm studies, 3 non-randomized controlled trials, and 1 case-control study. The oral administration of atenolol for the treatment of infantile hemangiomas resulted in an efficacy rate (complete remission rate) of 62% (95% CI 52%~71%). The incidence rate of adverse reactions related to the digestive system was 18% (95% CI 7%~30%), while that related to β2 receptor blockade was 4% (95% CI 2%~6%), central nervous system-related adverse reactions occurred at a rate of 10% (95% CI 5%~16%), the recurrence rate was 5% (95% CI 2%~9%). Egger’s test indicated that there was no significant publication bias in the efficacy rate, central nervous system-related adverse reaction rate, and gastrointestinal-related adverse reaction rate of oral atenolol treatment for infantile hemangiomas ( P>0.05). The sensitivity analysis for the efficacy rate, adverse reaction rate, and recurrence rate of oral atenolol treatment for infantile hemangiomas suggested that the result were stable and reliable. Conclusion:Oral administration of atenolol for the treatment of infantile hemangiomas demonstrates significant efficacy, fewer adverse reactions, and a low recurrence rate, making it a promising candidate as a reasonable alternative to oral propranolol for treating infantile hemangiomas.
5.Meta-analysis of the efficacy and safety of oral atenolol versus oral propranolol in the treatment of infantile hemangioma
Jianmin ZHENG ; Zhisheng ZHU ; Chaoyang WANG ; Shize ZHU
Chinese Journal of Plastic Surgery 2022;38(3):275-283
Objective:To explore the difference of the efficacy, safety and recurrence rate of oral atenolol compared with oral propranolol in the treatment of infantile hemangioma, so as to provide evidence-based medicine basis and reference for clinic.Methods:A comprehensive search was conducted on the English databases Web of Science, PubMed, Cochrane Library, Embase, U. S. National Library of Medicine Clinical Trials Registry Platform (https: //clinicaltrials.gov) and on the Chinese databases CNKI, CBM, VIP, Wanfang Data from January 2008 to June 2021, according to our defined inclusion and exclusion criteria, randomized controlled trials of oral atenolol versus oral propranolol in the treatment of infantile hemangioma were selected for performing meta-analysis, and the outcome indicators were treatment efficiency, incidence of adverse reactions, and recurrence rate. Meta-analysis was performed by using RevMan 5.3 software, and sensitivity analysis of the result was performed, and the main outcome indicators were tested for publication bias (Egger’s test) by using Stata 16 software.Results:Finally, 5 randomized controlled trials references were included. Our meta-analysis showed that there was no significant difference in the effective rate between oral atenolol and oral propranolol in the treatment of infantile hemangioma ( RR=0.93, 95% CI 0.84-1.02, P=0.110). There was a statistically significant difference in the overall incidence of adverse reactions ( RR=0.78, 95% CI 0.61-0.99, P =0.040), bronch-related and central nervous system related to β 2-blockade( RR=0.55, 95% CI 0.40-0.76, P<0.001) adverse reactions, which were lower in the atenolol group than in the propranolol group; there was a statistically significant difference in the recurrence rate ( RR=0.57, 95% CI 0.39-0.84, P=0.005), which was lower in the atenolol group than in the propranolol group. The sensitivity analysis showed that the result after the exclusion of any 1 study were less variable compared with the result of the previous analysis, and the conclusion obtained were unchanged, suggesting that the result of the meta-analysis were stable and reliable. The Egger’s test showed that P=0.502, which suggested that there was no obvious publication bias. Conclusions:In the treatment of infantile hemangioma, oral atenolol has equivalent efficacy compared with oral propranolol, with less overall incidence of adverse reactions (which can reduce the incidence of bronch-related and central nervous system adverse reactions) and lower recurrence rate.
6.Meta-analysis of the efficacy and safety of oral atenolol versus oral propranolol in the treatment of infantile hemangioma
Jianmin ZHENG ; Zhisheng ZHU ; Chaoyang WANG ; Shize ZHU
Chinese Journal of Plastic Surgery 2022;38(3):275-283
Objective:To explore the difference of the efficacy, safety and recurrence rate of oral atenolol compared with oral propranolol in the treatment of infantile hemangioma, so as to provide evidence-based medicine basis and reference for clinic.Methods:A comprehensive search was conducted on the English databases Web of Science, PubMed, Cochrane Library, Embase, U. S. National Library of Medicine Clinical Trials Registry Platform (https: //clinicaltrials.gov) and on the Chinese databases CNKI, CBM, VIP, Wanfang Data from January 2008 to June 2021, according to our defined inclusion and exclusion criteria, randomized controlled trials of oral atenolol versus oral propranolol in the treatment of infantile hemangioma were selected for performing meta-analysis, and the outcome indicators were treatment efficiency, incidence of adverse reactions, and recurrence rate. Meta-analysis was performed by using RevMan 5.3 software, and sensitivity analysis of the result was performed, and the main outcome indicators were tested for publication bias (Egger’s test) by using Stata 16 software.Results:Finally, 5 randomized controlled trials references were included. Our meta-analysis showed that there was no significant difference in the effective rate between oral atenolol and oral propranolol in the treatment of infantile hemangioma ( RR=0.93, 95% CI 0.84-1.02, P=0.110). There was a statistically significant difference in the overall incidence of adverse reactions ( RR=0.78, 95% CI 0.61-0.99, P =0.040), bronch-related and central nervous system related to β 2-blockade( RR=0.55, 95% CI 0.40-0.76, P<0.001) adverse reactions, which were lower in the atenolol group than in the propranolol group; there was a statistically significant difference in the recurrence rate ( RR=0.57, 95% CI 0.39-0.84, P=0.005), which was lower in the atenolol group than in the propranolol group. The sensitivity analysis showed that the result after the exclusion of any 1 study were less variable compared with the result of the previous analysis, and the conclusion obtained were unchanged, suggesting that the result of the meta-analysis were stable and reliable. The Egger’s test showed that P=0.502, which suggested that there was no obvious publication bias. Conclusions:In the treatment of infantile hemangioma, oral atenolol has equivalent efficacy compared with oral propranolol, with less overall incidence of adverse reactions (which can reduce the incidence of bronch-related and central nervous system adverse reactions) and lower recurrence rate.
7. Using the facial artery perforator-based nasolabial para-nasal advanced flap to repair the medial canthus and inner lower eyelid skin defects
Chaoyang WANG ; Xingong LIN ; Xianying ZHOU ; Shiquan WU ; Youyi WU ; Shize ZHU
Chinese Journal of Plastic Surgery 2019;35(1):49-52
Objective:
To discuss the surgical method and clinical effect of applying the facial artery perforator-based nasolabial para-nasal advanced flap to repair the medial canthus and inner lower eyelid skin defects.
Methods:
The advance nasolabial para-nasal perforator flap supplied by facial artery, was used to repair the medial canthus and inner lower eyelid skin defects, caused by dermatoma excision.
Results:
All 18 flaps completely survived. The detects in the medial canthus andinner lower eyelid, and the donor sites in the nasolabial fold were primary healed.The medial canthus and inner lower eyelid were recovery satisfactorily.The flaps were not bloated, and the contour and texture of flaps were similar to adjacent tissue, with no need of secondary repair.The donor site was successfully hidden in the nasolabial dermatoglyph.
Conclusions
Nasolabial para-nasal perforator flap is easily obtained, reliable in blood supply, and flexible in transfer. It has a wide range of movement and is easy to advance, so as to repair medial canthus andinner lower eyelid defect. With above advantages, this flap is worthy towidely popularize.
8.Effects of GSK-3βknockdown by RNA interference on formation of ke-loid in vitro
Yumei CAI ; Shize ZHU ; Weiqun YANG ; Mingmeng PAN ; Chaoyang WANG ; Wenyi WU
Chinese Journal of Pathophysiology 2017;33(1):154-160
AIM: To study the suppressive effect of glycogen synthase kinase-3β( GSK-3β) knockdown by RNA interference on the formation of keloid .METHODS:Human keloid fibroblasts ( KFB) in vitro were transfected with 3 pairs of specific GSK-3βsmall interfering RNA (siRNA).The best siRNA to inhibit the GSK-3βexpression in human KFB was screen by RT-PCR and Western blot .The expression of GSK-3βand related proteins at mRNA and protein levels in the KFB was determined by RT-PCR and Western blot .RESULTS: The GSK-3βsiRNA1434 remarkably inhibited the expression of GSK-3βat mRNA and proteins levels in the human KFB .After transfection with GSK-3βsiRNA, the protein levels of β-catenin, p-GSK-3β, Wnt2 and cyclin D1 were all decreased.KFB growth became slow.With the extension of time, the inhibition of cell growth increased , and the cell doubling time was significantly delayed .CONCLUSION:siRNA targeting GSK-3βefficiently knocks down the expression of GSK-3βin the human KFB, and inhibits the activation of Wnt signaling pathway , thus inhibiting the growth of keloid .GSK-3βmay be a potential therapeutic target for keloid .
9.Expression of WWOX and C-JUN in keloid
Yumei CAI ; Weiqun YANG ; Shize ZHU ; Wenyi WU ; Shaohua WANG ; Yongfeng LI
Chinese Journal of Medical Aesthetics and Cosmetology 2014;20(6):447-450
Objective To study the expression of WWOX and C-JUN in keloid and to approach their role and mechanism in the pathogenesis of keloid.Methods Immunohistochemical SP methods were used with computer pathological image analysis.Western blot and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of WWOX and C-JUN in keloid and normal skin with statistical analysis.Results In keloid,the expression of WWOX protein was located in the cytoplasm of fibroblasts,and the expression of WWOX protein and its mRNA decreased,with significantly statistical difference (P<0.05) compared to normal skin in the control group; the expression of C-JUN protein was located in the cell nucleus and cytoplasm of fibroblasts,with increased expression of C-JUN protein and its mRNA,with significantly statistical difference (P<0.05) in comparison to normal skin in the control group.The expression of both was negative correlation (r=-0.626,P<0.01).Conclusions Both WWOX with low expression and C-JUN with high expression are keloid-related genes,having significantly negative correlation between them,which may be one of the mechanisms for the keloid formation.It indicates that the WWOX protein may be an inhibitory factor to the expression of C-JUN protein,and the genes may play a major role in the pathogenesis of keloiod through fibroblasts.
10.Expressions and significance of ZNF217 in papillary thyroid carcinoma
Liting ZHANG ; Wenyi WU ; Zhaoyang WANG ; Zhongxin HUANG ; Shize ZHU
Chinese Journal of Endocrinology and Metabolism 2013;29(9):779-781
Objective To investigate the expression and clinical significance of zinc finger gene 217 (ZNF217) in human papillary thyroid carcinoma.Methods The expressions of ZNF217 mRNA and protein were detected by quantitative realtime PCR and Western blot in papillary thyroid carcinoma tissues (n =20) and adjacent normal tissues (n =20),and the data were analyzed.Results The expressions of ZNF217 mRNA and protein in papillary thyroid carcinoma were significantly higher than those in adjacent normal tissues (96.72 ± 44.19 vs 4.86 ±3.55,0.994 ± 0.172 vs 0.195 ± 0.061,both P<0.01),being higher in the papillary thyroid carcinoma with capsule invasion compared with that without capsule invasion (P<0.01).The expressions of ZNF217 mRNA and protein in papillary thyroid carcinoma were not related to gender,age,tumor size,TNM stage or lymph node metastasis (all P>0.05).Conclusions The overexpression of ZNF217 may be associated with the oncogenesis and progress of papillary thyroid carcinoma and capsule invasion,and thus is expected to become a new target for prevention and treatment of papillary thyroid carcinoma.

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