1.Construction of mouse MyoD vector and its expression in Escherichia coli
Ruifeng QIN ; Xiaoming GU ; Xuerong XING ; Tianqiu MAO
Journal of Practical Stomatology 1995;0(04):-
Objective:To identify the cDNA gene of mouse MyoD by restriction enzyme analysis, and to express the gene in Escherichia coli (E coli) using a protein expression vector.Methods: After the cDNA gene of mouse MyoD had been amplified and identified,it was inserted into expression vector pBV220 in which exogenous gene was controlled by R RP L promoters.The recombinant plasmid pBV-my was transformed into E coli DH5? and the bacteria were induced at 42 ℃ to express encoded protein.Results:The cDNA of mouse MyoD was sequenced correctly.When the engineered bacteria had been induced an anticipated 55 ku protein band from the bacteria was observed on SDS-PAGE gel and amounted to 30% of total bacterial protein. Conclusion:The cDNA of mouse MyoD has been successfully coloned and efficiently expressed in E coli.
2.Analysis of ABO and RhD blood group detection results among voluntary blood donors in Huainan of Anhui province
Hui ZHANG ; Tao ZHOU ; Xing CHENG ; Yujiao SHUI ; Xun FANG ; Xuerong LIU ; Dong LI
Chinese Journal of Primary Medicine and Pharmacy 2023;30(8):1216-1219
Objective:To analyze the distributions of ABO and RhD blood groups by analyzing the basic data of blood group detection among voluntary blood donors in Huainan in 2021, to provide data support for blood recruitment, clinical use of blood, and emergency guarantee of rare groups of blood.Methods:ABO and RhD blood groups of 24 484 voluntary blood donors eligible for blood donation in 2021 were detected using the Metis150-8 automatic blood group analyzer, manual saline method, antihuman globulin method, and manual polybrene test. The blood group results were statistically analyzed.Results:Among 24 484 voluntary blood donors in Huainan in 2021, A blood group accounted for the highest proportion (7 463 cases, 30.48%), followed by O blood group (7 444 cases, 30.40%) and B blood group (7 056 cases, 28.82%), and the last was AB blood group (2 521 cases, 10.30%). A total of 143 cases of RhD-negative blood were detected, and the negative frequency of RhD was 0.58% (143/24 484). Among the RhD-negative blood samples, 43 cases of type A, 41 cases of type B, 46 cases of type O, and 13 cases of type AB were RhD-negative, accounting for 30.07%, 28.67%, 32.17%, and 9.09%, respectively. There was no statistical difference in the detection rate of Rh-negative blood among different ABO blood groups in Huainan ( χ2 = 0.36, P = 0.948). The ABO blood group distribution of voluntary blood donors in Huainan in 2021 was not identical to those of voluntary blood donors in Guangzhou, Yueyang, Xinjiang Bozhou, Zhangzhou, and Liuzhou. The proportion of type A blood donors in Yueyang was higher than those in other regions. The proportions of type B blood donors in Huainan, Xinjiang Bozhou, and Zhangzhou were higher than those in other regions. The proportion of type O blood donors in Liuzhou was higher than those in other regions. The proportions of type AB blood donors in Huainan and Xinjiang Bozhou were higher than those in other regions. Conclusion:The distributions of ABO and RhD blood groups among voluntary blood donors in Huainan region have certain regional characteristics. Central blood banks and medical institutions should reasonably store and supply blood according to the blood collection from voluntary blood donations and the needs of clinical transfusion, to prevent the occurrence of situations such as blood expiration and waste.
3.In vitro anti-influenza A virus H3N2 activity of lithium chloride
Hongkai ZHANG ; Jia ZANG ; Yanshi WU ; Yueping XING ; Zefeng DONG ; Xuerong YA ; Qiang SHEN
Chinese Journal of Experimental and Clinical Virology 2024;38(5):539-546
Objective:To analyze the activity of lithium chloride (LiCl) against influenza virus A (H3N2) in human non-small cell lung cancer cells (A549).Methods:Different concentrations of LiCl were incubated with A549 cells, and the cytopathic effect (CPE) was observed after 24 hours, and the effect of LiCl on cell activity was determined by CCK-8 method. After H3N2 (MOI=1) infected A549 cells, different concentrations of LiCl were added and incubated for 24 hours, and the viral load was measured by real time/reverse transcription quantitative polymerase chain reaction (RT-qPCR), and the CPE was observed, and the viral titer was determined. Different concentrations of LiCl were incubated with A549 at 37 ℃ and 5% CO 2 for 2 hours, virus was added and incubated for 24 hours, and the viral load was determined by RT-qPCR. LiCl, H3N2 and A549 were incubated at 4 ℃ for 1 hour, 35 ℃, 5% CO 2 for 24 hours, and viral load was determined by RT-qPCR. H3N2 and A549 were incubated at 4 ℃for 1 hour, then different concentrations of LiCl were added, incubated at 35 ℃ with 5% CO 2 for 24 hours, and the viral load was determined by RT-qPCR. After H3N2 infected A549 cells, different concentrations of LiCl were added and incubated for 24 hours, and the viral RNA load and viral titer of the supernatant and cells were measured, respectively, and then the corresponding ratios of the supernatant and the cells were calculated. After H3N2 (MOI=10) and BV (MOI=1) infected A549 cells, different concentrations of LiCl were added for 24 h, and the viral load was determined by RT-qPCR. Results:When the concentration of LiCl was<50 mmol/L, the cell viability of A549>90%. Different concentrations of LiCl could significantly reduce the viral load of H3N2 ( P<0.000 1), and the CPE of the LiCl treatment group was more dose-dependent than that of the control group. LiCl did not inhibit viral replication by affecting the cell itself; Different concentrations of LiCl significantly inhibited the entry of H3N2 into A549 ( P<0.000 1), and also had a certain inhibitory effect on the adsorption of A549 cells ( P<0.1). LiCl did not affect the assembly and release of H3N2 ( P>0.05), and it was also found that LiCl had a broad spectrum of antiviral effects against multiple influenza virus strains ( P<0.000 1). Conclusions:LiCl may exert antiviral effect by inhibiting the adsorption and entry of H3N2 into A549 cells and the replication of H3N2 in A549 cells, which provides a data reference for the prevention and treatment of viral infection by LiCl.
4.Investigation and Analysis of the Satisfaction and Confidence of the Students Majoring in Non-medical Interdisciplinary Subject in Our University
Hongyan WU ; Long WANG ; Xiong CHEN ; Minghu CEN ; Xing YANG ; Yan ZHANG ; Lei TANG ; Xuerong RAN ; Jia SUN
China Pharmacy 2017;28(27):3750-3754
OBJECTIVE:To improve the education quality of medical school,so as to provide the excellent human resources for the field of health.METHODS:2011-2014 grades of students majoring in pharmaceutical administration,pharmacy (marketing direction) and other non-medical interdisciplinary subjects were selected from our school to complete the questionnaire which conrained the questions about satisfaction and confidence for the major.The results of investigation were analyzed statistically.RESULTS:Totally 2 668 questionnaires were sent out,and 2 435 were recycled with effective recovery of 91.27%.Half of the students majoring in pharmaceutical administration or pharmacy (marketing) etc.were switched from other majors,and 38.40%(935 number) of the students wanted to change the major.The major of clinical,nursing and anesthesiology were most preferred by the students.Only 15.52% (378 number),26.32% (641 number) of the students thought that the school paid enough attention to them or had a positive attitude to their major,respectively.Students majoring in pharmacy (marketing) and health services administration had the lowest satisfaction about their major and the attention paid by the school (P<0.001).With the growth of grade,the satisfaction of students in various aspects had declined (P<0.001).Only 44.72% (1 089 number),34.25% (834 number) of the students had positive attitude towards the study or were optimistic about the future,respectively.With the growth of the grade,students' satisfaction in all aspects had declined (P<0.001).Compared to other students,freshman had more positive attitude towards the study or were more optimistic about the future (P<0.001).Besides,33.65% (491 number)of the students planning to work immediately after graduation lacked confidence to find a good job.CONCLUSIONS:The lower degree of satisfaction and confidence are found in the students majoring in non-medical interdisciplinary subject in the medical school,and this should be treated seriously by the school.