2.Genetic recombinant prokaryotic expression of augmenter of liver regeneration and preparation of polyclonal antibodies in rats
Zhengfang LIU ; Jianming WANG ; Lan WANG ; Xiaoyun ZENG ; Ling XIONG ; Zhixiu LUO ; Junyi WU
Chinese Journal of Tissue Engineering Research 2010;14(11):1943-1947
BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.
3.Regulatory Effects of miRNA-31 on LATS2 and Cardiomyocyte Hypertrophy in Rat's Cardiomyocytein vitro
Junyi ZENG ; Wan ZHANG ; Lu DING ; Yunfeng WEI ; Zeqi ZHENG ; Tong WEN ; Yongnan FU
Chinese Circulation Journal 2017;32(2):177-182
Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.
4.Elastic registration method to compute deformation functions for mitral valve.
Jinyu YANG ; Wan ZHANG ; Ran YIN ; Yuxiao DENG ; Yunfeng WEI ; Junyi ZENG ; Tong WEN ; Lu DING ; Xiaojian LIU ; Yipeng LI
Journal of Biomedical Engineering 2014;31(5):1135-1138
Mitral valve disease is one of the most popular heart valve diseases. Precise positioning and displaying of the valve characteristics is necessary for the minimally invasive mitral valve repairing procedures. This paper presents a multi-resolution elastic registration method to compute the deformation functions constructed from cubic B-splines in three dimensional ultrasound images, in which the objective functional to be optimized was generated by maximum likelihood method based on the probabilistic distribution of the ultrasound speckle noise. The algorithm was then applied to register the mitral valve voxels. Numerical results proved the effectiveness of the algorithm.
Algorithms
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Heart Valve Diseases
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diagnostic imaging
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Humans
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Likelihood Functions
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Mitral Valve
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diagnostic imaging
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pathology
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Patient Positioning
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Probability
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Ultrasonography
5.Analysis of the Relevance between t-BHP Induced Apoptosis and STATs mRNA Expression in HEI-OC1 Cells
Zihao ZENG ; Xuan LUO ; Yuheng YAO ; Junyi WANG
Journal of Audiology and Speech Pathology 2018;26(2):162-166
Objective To study the biological effects of (Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription (STATS) of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations (0,20,40,60μM) of(Tert-Butyl hydroperoxide,T-BHP) built the apoptotic HEI-OC1 cells model while 0 μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60 μM) were 3.9%,7.4%,32.0%,and 91.2%,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STATS mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly (F=5 534.302,P<0.01;F=146.038,P<0.01;F=685.929,P<0.01;F=516.11,P< 0.01).No statistically significant differences were found among the poisoning concentration groups (P>0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly (F=1 259.148,P< 0.01).The STAT2 mRNA expression level in 20 μM contamination group decreased while 40,60 μM contamination group increased significantly (P<0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.
6. The influence of STAT2 siRNA inhibition on tert-butyl hydroperoxide-induced apoptosis in HEI-OC1 cells
Shoukai LIN ; Zihao ZENG ; Min ZHANG ; Junyi WANG
China Occupational Medicine 2019;46(05):581-585
OBJECTIVE: To analyze the influence of signal transducers and activators of transcription 2(STAT2) on the apoptosis of oxidative stress-induced HEI-OC1 hair cells. METHODS: With random number table method,HEI-OC1 hair cells were divided into interference group and non-interference group. The artificial synthesis design targeted STAT2 small interfering RNA(siRNA) and opti-modified Eagle medium(opti-MEM) were added into the interference group, and only the opti-MEM was added into the non-interfere group. The 0, 20, 40, 60 μmol/L of tertiary butyl peroxide hydrogen(t-BHP) was added into the interference and non-interference group. Western blotting was used to detect the relative expression of STAT2 and cysteine protease(caspase)-3. The apoptotic rate of hair cells was detected by flow cytometry. RESULTS: In the interference group and non-interference group, the apoptotic rate of HEI-OC1 cells and the relative expression level of STAT2, Caspase-3 increased with the dose of t-BHP(P<0.01). The apoptotic rate of HEI-OC1 cells and the relative expression level of Caspase-3 in the interference group with 20, 40, 60 μmol/L of t-BHP were higher than that of the 20, 40 μmol/L group(P<0.01). The apoptotic rate of HEI-OC1 cells in the interference group was higher than those in the non-interference group with the same dose(P<0.01), while the relative expression level of STAT2 was lower than those in the non-interference group with the same dose(P<0.01). CONCLUSION: Inhibiting oxidative stress and STAT2 expression in hair cells can lead to aggravation of apoptosis. STAT2 has anti-apoptotic effect.
7.Efficacy of Lenvatinib Combined with Anti–PD-1 Antibodies Plus Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus: A Retrospective, Multicenter Study
Xiangye OU ; Junyi WU ; Jiayi WU ; Yangkai FU ; Zhenxin ZENG ; Shuqun LI ; Yinan LI ; Deyi LIU ; Han LI ; Bin LI ; Jianyin ZHOU ; Shaowu ZHUANG ; Shuqun CHENG ; Zhibo ZHANG ; Kai WANG ; Shuang QU ; Maolin YAN
Cancer Research and Treatment 2024;56(4):1207-1218
Purpose:
The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT.
Materials and Methods:
This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles.
Results:
During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively.
Conclusion
Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.
8.Efficacy of Lenvatinib Combined with Anti–PD-1 Antibodies Plus Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus: A Retrospective, Multicenter Study
Xiangye OU ; Junyi WU ; Jiayi WU ; Yangkai FU ; Zhenxin ZENG ; Shuqun LI ; Yinan LI ; Deyi LIU ; Han LI ; Bin LI ; Jianyin ZHOU ; Shaowu ZHUANG ; Shuqun CHENG ; Zhibo ZHANG ; Kai WANG ; Shuang QU ; Maolin YAN
Cancer Research and Treatment 2024;56(4):1207-1218
Purpose:
The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT.
Materials and Methods:
This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles.
Results:
During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively.
Conclusion
Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.
9.Efficacy of Lenvatinib Combined with Anti–PD-1 Antibodies Plus Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus: A Retrospective, Multicenter Study
Xiangye OU ; Junyi WU ; Jiayi WU ; Yangkai FU ; Zhenxin ZENG ; Shuqun LI ; Yinan LI ; Deyi LIU ; Han LI ; Bin LI ; Jianyin ZHOU ; Shaowu ZHUANG ; Shuqun CHENG ; Zhibo ZHANG ; Kai WANG ; Shuang QU ; Maolin YAN
Cancer Research and Treatment 2024;56(4):1207-1218
Purpose:
The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT.
Materials and Methods:
This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles.
Results:
During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively.
Conclusion
Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.
10.Efficacy of Lenvatinib Combined with Anti–PD-1 Antibodies Plus Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus: A Retrospective, Multicenter Study
Xiangye OU ; Junyi WU ; Jiayi WU ; Yangkai FU ; Zhenxin ZENG ; Shuqun LI ; Yinan LI ; Deyi LIU ; Han LI ; Bin LI ; Jianyin ZHOU ; Shaowu ZHUANG ; Shuqun CHENG ; Zhibo ZHANG ; Kai WANG ; Shuang QU ; Maolin YAN
Cancer Research and Treatment 2024;56(4):1207-1218
Purpose:
The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT.
Materials and Methods:
This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles.
Results:
During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively.
Conclusion
Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.