1.A novel method to evaluate myocardial injury: cloning of anti-cTnI murine antibody Fab fragment and DNA sequence analysis
Yanyan LI ; Di YANG ; Jiannan LIU ; Zhiping BIAN ; Jindan XU ; Xiangjian CHEN ; Chunrang GU ; Jinan ZHANG
Chinese Journal of Tissue Engineering Research 2005;9(19):213-215
BACKGROUND: To apply mouse anti-human cTnI monoclonal antibody as the drug vector in the treatment and diagnosis of myocardial injury, it is important to degrade the immunity of murine antibody and overcome human anti-mouse reaction. Humanization has been applied as an attempt to resolve this problem.OBJECTIVE: To clone murine anti-cTnI Fab fragment and analyse the nucleotide and deduced amino acid sequences.DESIGN: Single sample study.SETTING: An institute of cardiovascular disease under a medical university-affiliated hospitalMATERIALS: The study was conducted in the Institute of Cardiovascular Diseases, First Affiliated Hospital of Nanjing Medical University from January 2003 to May 2004. The hybridoma cell line JS200202 which secrets the anti-cTnI monoclonal antibody was provided by Institute of Cardiovascular Disease, First Affiliated Hospital of Nanjing Medical University.METHODS: IgG heavy chain primers and κ light chain primers of amplified mouse were designed. Total RNA was extracted from hybridoma cells which secrete cTnI. Reverse transcription polymerase chain reaction(RT-PCR) was amplified. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences.MAIN OUTCOME MEASURES: Heavy chain Fd segment and κ light chain gene sequence and its subgroups.RESULTS: A band of approximate 700 and 800 base pairs were amplified using IgG heavy chain primers and κ light chain primers respectively. Sequence analysis indicated that the deduced amino acid sequences were in consistent with the characterization of the amino acid in the murine IgGl Fab fragment(GenBank accession NO AY484430, AY484431; Protein Bank accession NO AAR83243, AAR83244).CONCLUSION: A complete murine anti-cTnI Fab fragment was obtained in this study, which may provide basis for the production of the chimeric anti-cTnI antibody.
2.Effect of low-intensity extracorporeal shock wave on the treatment of chronic prostatitis in rats
Chen JIN ; Zichen BIAN ; Zhengyao SONG ; Zhiping WANG ; Bin FENG ; Li ZHANG ; Zongyao HAO ; Song FAN ; Cheng YANG ; Yi LIU ; Chaozhao LIANG
Chinese Journal of Urology 2020;41(10):779-783
Objective:To explore the therapeutic effect of low-intensity extracorporeal shock wave (Li-ESW) in model rats with chronic prostatitis and its optimal parameters.Methods:From April to August 2019, 90 healthy male SD rats aged 8 weeks were randomly divided into control group (group C, n=15), model group (group M, n=15) and shock wave treatment group (group T, n=60), which were treated with Li-ESW after being modeled successfully. The rats in group T were supine and treated with focused Li-ESW, and the probe was placed above the anatomical position of the prostate. Treatment scheme was 3 Hz of frequency, 500 pulses, and once a week for 4 weeks. Group T was divided into group T1(0.09 mJ/mm 2), T2(0.20 mJ/mm 2), T3(0.30 mJ/mm 2) and T4(0.40 mJ/mm 2) according to energy flux density, with 15 rats in each group. Before the establishment of the model, the rats in each group were tested with von Frey fiber of 2 g, 4 g and 6 g at the scrotum respectively, and the positive reaction was recorded. There was no significant difference between the three groups ( P>0.05). After one week, the rat model with CP was established by injecting 3% carrageenan into the bilateral lobes of the prostate under anesthesia. And one week after the modeling was completed, the rats in each group underwent von Frey test again to measure pain and evaluate the modeling effects. Group T was then treated once a week for a 4-week period. During the treatment, von Frey test was performed before each treatment to evaluate the treatment effect of the previous week. After the von Frey test for one, two and four weeks of treatment, 5 rats were sacrificed in each group, and the bilateral lobes of the prostate were dissected under aseptic conditions for paraffin inclusion and HE staining. Result:The positive reaction in group M and each group T was significantly more than those in group C ( P<0.001) one week after the modeling was completed, and there was no significant difference between group M and group T. The number of positive reactions in group T2 was lower than that in group M at each time point after treatment ( P<0.05), and it is also less in group T1, T3 and T4 were than that in group M after 3 weeks, 3 and 4 weeks and 2 weeks respectively( P<0.05). In group T2, the number of positive reactions began to decrease after 1 week of treatment, and reached the lowest after 4 weeks, with significant difference in pain measurement results at different time points ( P<0.05). The results of pathological examination showed that there were more inflammatory cells in prostatic stroma and disordered arrangement of epithelial cells in group M than that in group C. The pathological scores of group M, T1, T2, T3 and T4 were 8, 7, 4, 6, 9 after 1 week treatment, 8, 5, 3, 4, 7 points after 2 weeks treatment, and 7, 3, 2, 4, 7 points after 4 weeks treatment, respectively. Conclusions:Low-intensity extracorporeal shock wave treatment had a significant effect on the improvement of symptoms in model rats with chronic prostatitis. Under the scheme of frequency 3 Hz, 500 pulses, once a week for 4 weeks, the optimal parameters is 0.20 mJ/mm 2.
3.Regional and demographic differences on passive smoking among non-smokers aged 30-79 years in 10 regions of China
Qi LIU ; Jiachen LI ; Huaidong DU ; Weihua CAO ; Jun LYU ; Yu GUO ; Zheng BIAN ; Zhiping LONG ; Pei PEI ; Junshi CHEN ; Canqing YU ; Zhengming CHEN ; Liming LI
Chinese Journal of Epidemiology 2020;41(10):1668-1673
Objective:To describe the regional and demographic differences on passive non-smokers from 10 regions involved in the China Kadoorie Biobank (CKB) study.Methods:Detailed information regarding passive smoking behaviors related to 317 486 non-smokers who were 30-79 years old from the 10 study regions were gathered and analyzed.Results:Following the standardization of the 2010 China national population, the prevalence rate of passive smoking was 56.7 %, and the prevalence rate of living with smokers was 66.5 % among the Chinese adults. Both of the aforementioned rates were higher in rural than in urban areas. Meanwhile, the regional distribution of weekly passive smoking frequency and cumulative duration of passive smoking per week and cumulative duration of passive smoking per day were significantly different. The cumulative passive smoking duration per week increased along with the weekly frequency in people living in urban areas. Among women, the weekly passive smoking frequency was the highest, and the cumulative durations per week and per day appeared the lowest in Hunan, opposite to the situation in Henan. The prevalence of passive smoking among participants living with smokers was 2.27 times (95 %CI: 2.24 -2.29) of those who were not and the association appeared stronger in women ( OR=2.61, 95 %CI: 2.58-2.64) but not in men ( OR=1.01, 95 %CI: 0.95-1.06). Almost all the indicators seemed higher in women than those in men, except for the cumulative duration per day. Furthermore, these indicators appeared higher among those who were at younger age or with less education. The prevalence rates of passive smoking and living with smokers were lower but the cumulative duration per day was higher among those with lower household income. And the two rates were higher in married women and lower in married men, as compared to their counterparts. Conclusion:Regional and demographic differences in passive smoking were noticed among study population of CKB in the 10 regions.