1.Accurate determination of the whole genome sequencing and open reading frames composition of non-replicating Tiantan strain of vaccinia virus based on novel long read sequencing platform
Zhongxian ZHANG ; Changcheng WU ; Han LI ; Shuting HUO ; Li ZHAO ; Yao DENG ; Xin MENG ; Roujian LU ; Houwen TIAN ; Wenling WANG ; Baoying HUANG ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2024;44(6):502-509
Objective:To accurately ascertain the whole genome sequencing and the composition of open reading frames (ORFs) of non-replicating Tiantan strain of vaccinia virus (NTV) using next-generation long-read sequencing technology.Methods:NTV, obtained from our laboratory stock, was amplified and purified on chicken embryo fibroblast cells(CEFs), and the full-length genomic nucleic acid of NTV was extracted. The PacBio HiFi sequencing platform was utilized for de novo assembly to obtain the complete genomic sequence of NTV. Using a homology annotation strategy, we identified its ORF composition and compared it with known non-replicating vaccinia virus strains. Results:The total length of NTV′s genome was 171 729 bp, with a GC content of 33%. Its unique inverted terminal repeat (ITR) region comprised hairpin structures, two tandem repeat regions, and three non-repeat regions. NTV contained 166 ORFs, with major differences observed in the ITR and its surrounding regions when compared to MVA-BN and NYVAC. These three strains shared a common set of 138 ORFs. NTV encoded six unique ORFs related to virus evasion of host antiviral response.Conclusions:This study accurately determines the whole genome sequencing and ORFs composition of NTV, and reveals its similarities and differences with other replication-deficient vaccinia virus strains, which pave a way for the development and application of the next generation of monkeypox vaccines and novel viral vectors.
2.Establishment of CRISPR/Cas12a-based molecular detection method for monkeypox virus
Meihui LUO ; Li ZHAO ; Changcheng WU ; Roujian LU ; Ruhan A ; Baoying HUANG ; Yao DENG ; Jiao REN ; Huijuan WANG ; Fei YE ; Wen WANG ; Houwen TIAN ; Wenling WANG ; Wenjie TAN
Chinese Journal of Experimental and Clinical Virology 2023;37(2):193-200
Objective:To develop a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a-based nucleic acid assay for monkeypox virus with high specificity and sensitivity.Methods:RAA primers and CRISPR RNA (crRNA) were designed based on the known conserved regions of the monkeypox virus gene and synthesized, and specific crRNAs were screened using fluorescence detection. The sensitivity and specificity of the detection system were evaluated.Results:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was developed with a sensitivity of 2.5 copies/reaction and high specificity without cross-reactivity with ectromelia virus and vaccinia virus.Conclusions:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was established, which would provide a powerful tool for efficient, rapid and specific detection of monkeypox virus.
3.Construction of the evaluation index system for the core competence of preventive medicine graduates to respond to sudden biological public health events
Chuancheng WU ; Xiaoying LIN ; Shuangfeng YANG ; Yu JIANG ; Baoying LIU
Chinese Journal of Medical Education Research 2022;21(3):277-280
Objective:To construct a set of evaluation index system for the core ability of preventive medicine graduates to respond to sudden biological public health events.Methods:Research methods such as in-depth interviews and group discussions were used to determine the core competence structure, the Delphi method was used to conduct two rounds of expert consultation, and the index system and its weights were finally determined based on the consultation results of 21 experts on the system. Excel and SPSS 21.0 were used for data collation and statistical analysis.Results:The positive coefficient of expert consultation was greater than 80%, the authority coefficient was greater than 0.7, and the average was 0.906. The final index system was composed of 4 first-level indicators, 10 second-level indicators, and 39 third-level indicators.Conclusion:The evaluation index system can provide a reference for exploring effective ways to improve the core competence of graduates.
4.Death Characteristics of Digestive System Cancer in Xianyou County, Fujian Province, 2014-2018
Yuhang ZHOU ; Shuangfeng YANG ; Baoying LIU ; Chuancheng WU
Cancer Research on Prevention and Treatment 2021;48(1):66-70
Objective To analyze the cancer mortality and epidemiological characteristics of digestive system cancer in Xianyou County, Fujian Province, and to provide a scientific evidence for formulating preventive measures for digestive system cancer. Methods Excel and SPSS software were used to collect the monitoring data of digestive system cancer death in Xianyou County from 2014 to 2018, and then analyzed the mortality, the standardized rate (according to the Chinese 2000 population census) and the cause of death. Results From 2014 to 2018, the deaths of malignant tumors in Xianyou County accounted for 35.00% of all deaths. Among them, the four common digestive system malignant tumors (gastric cancer, esophageal cancer, liver cancer and colorectal cancer) accounted for 62.44% of malignant tumors. Gastric cancer ranked first and accounted for 27.36%. Average annual rate of crude mortality and standardized mortality of malignant tumor in the digestive system were 108.93/105 and 89.34/105, respectively, and the ratio of male to female was 2.27 to 1. Significant differences were found between town and village, as the terrain gets lower, the death rate goes down gradually. Compared with the data of 2011-2012, the standardized death rate of gastric cancer, esophageal cancer and liver cancer were reduced by 29.71%, 23.92% and 12.06%, respectively, while the rate of colorectal cancer was increased by 28.80% and the average annual growth rate was 7.83%. Conclusion Although the death rates of gastric cancer, esophageal cancer and liver cancer have decreased significantly, the malignant tumor of the digestive system is still the main disease that causes the death of residents in Xianyou County, Putian City. Therefore, it is necessary to strengthen the education of health in order to achieve the early discovery, diagnosis and treatment.
5.Ross procedure to cure aortic valve disease in children with expended polytetrafluoraethylene(ePTFE) artificial valve for right ventricular outflow tract reconstruction
Pengcheng WANG ; Cheng ZHANG ; Suixin LIANG ; Baoying MENG ; Huaipu LIU ; Junrong HUANG ; Wenzhi WU ; Keye WU ; Yiqun DING
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(12):711-716
Objective:To review and analyze the clinical experience of children with aortic valve stenosis and/or insufficiency treated with autologous pulmonary valve for aortic valve replacement procedure(Ross operation) with ePTFE artificial valve for right ventricular outflow tract reconstruction.Methods:From 2015 to 2020, 8 cases of aortic stenosis and/or aortic insufficiency treated by Ross operation in our center were collected, with an age of 0.5-13.2 years old. 4 cases of aortic stenosis were diagnosed preoperatively, 3 cases of aortic stenosis with aortic insufficiency, and 1 case of infective endocarditis involving the aortic valve. The operation was carried out in three steps: Harvest autologous pulmonary valve; the diseased aortic valve was resected and autologous pulmonary valve was transplanted to the aorta by aortic root transplantation; the right ventricular outflow tract was reconstructed by a handmade ePTFE artificial flap blood vessel.Results:In 6 cases, the right ventricular outflow tract was reconstructed by hand-sewn ePTFE trileaflets, and artificial univalve in 2 cases, no death occurred during operation; all patients were cured and discharged. The patients were followed up for 1 to 36 months, with mean of(12.63±12.19) months. There was no long-term death or valvular complications. During follow-up echocardiography indicated 1 case of moderate aortic regurgitation, 1 case of mild-moderate regurgitation, and moderate regurgitation was found in 2 patients with artificial single pulmonary valve. For the remaining patients, they were mild aortic regurgitation, and a trivial or mild pulmonary artery regurgitation with hand-sewn three-leaflets ePTFE artificial vessel; All patients were followed up at the last time with a peak pressure of(6.63±3.46) mmHg(1 mmHg=0.133 kPa) across the aortic valve. The left ventricular outflow tract and aortic annulus shrank slightly after surgery(the diameter of one patient with Ross-Konno operation increased), but the annulus diameter increased with age. There was no need for further intervention.Conclusion:The Ross operation is safe for the treatment of aortic valve disease, it has good hemodynamic effect, and the autologous pulmonary artery has growth potential, especially suitable for children and young patients. Hand-sewn ePTFE with trileaflet vessels for reconstruction of right ventricular outflow tract performs well in anti-regurgitation function in the short term or may be used as a replacement material for the homograft/heterograft vessel, but longer follow-up and more cases are needed.
7. Associations of parental thalassemia with preterm birth and low birth weight
Xiaochun HUANG ; Xiaoqiang QIU ; Xiaoyun ZENG ; Shun LIU ; Fanglian WEI ; Yanning LI ; Tao LIU ; Yanan WU ; Baoying FENG ; Qunjiao JIANG ; Dongping HUANG
Chinese Journal of Epidemiology 2019;40(5):596-600
Objective:
To investigate the association between the preterm birth and low birth weight and parental thalassemia.
Methods:
Pregnant women and their husbands receiving prenatal examination in local hospitals or maternal and child health centers in Jingxi and Debao in Guangxi from January to December 2017 were selected as study subjects. A total of 758 pregnant women with pregnancy outcomes and their husbands, who were both or alone diagnosed with thalassemia through thalassemia gene detection, were selected as case group and 758 pregnant women with pregnancy outcomes and their husbands, who were negative in thalassemia gene detection and hemoglobin electrophoresis test were selected as control groups. The case group were further divided into mother group, father group and both mother and farther group. Clinical and pregnancy outcome data of the study subjects were collected for the analysis on the association between parental thalassaemia and preterm birth or low birth weight by the independent sample
8. Design of self-evaluation scale on basic ability in graduates of preventive medicine major and its reliability and validity test
Shuangfeng YANG ; Chuancheng WU ; Xiaoying LIN ; Wenchang ZHANG ; Huangyuan LI ; Yueping LI ; Baoying LIU
Chinese Journal of Medical Education Research 2019;18(9):952-956
Objective:
To design the self-rating scale on basic ability in graduates of preventive medicine major, and test its reliability and validity, providing a reliable tool for evaluating the basic ability of graduates.
Methods:
On the basis of constructing the Evaluation Index System on Basic Ability in Graduates of Preventive Medicine Major, the three-level indexes were converted to specific questions. Self-evaluation scale including 58 items, 22 sub-dimensions and 8 main dimensions were finally designed after pre-survey and expert argumentation. A total of 205 graduates of preventive medicine major from a medical college and 9 experts in the field of public health were selected to completed the questionnaire. Scale's reliability was tested by applying internal consistency reliability and composite reliability; it's validity was tested by applying content validity and confirmatory factor analysis.
Results:
Scale's α coefficient of internal consistency reliability was 0.976; α coefficient and combination reliability of eight main dimensions were both higher than 0.8; the content validity index S-CVI of the scale was 0.94, the content validity index I-CVI of the item level was 0.78 to 1.00; the average variance extraction amount (AVE) of the eight main dimensions was higher than 0.50. Fitting of the structural model with the item used as observation indexes was reasonable (
10.The effect of inhibition of hiwi expression by RNAi on triple negative breast cancer cells MDA-MB-231 through inducing cell apoptosis
Baoying CHEN ; Haibing WU ; Yuanqi ZHANG ; Shengchao HUANG ; Ying ZHANG ; Jianwen LI ; Yuzhou WANG ; Wangting ZHENG ; Luowu WU
The Journal of Practical Medicine 2017;33(9):1385-1389
Objective To investigate the effect of interfering Hiwi gene on the apoptosis of MDA-MB-231 cells. Methods The mRNA and protein expression of Hiwi mRNA and its target protein were analyzed by qRT-PCR and Western Blot after transfection. MDA-MB-231 cells were divided into 6 groups according to the experimental design. Interference effects were screened as siRNA interference group (Hiwi10330 group), and then divided into 3 groups according to the experimental design: interference group, negative control group/NC, blank control group/Blank. The cell apoptosis rate was detected by flow cytometry after transfection. Results The expression of mRNA in the interference group was significantly lower than that in the siRNA group (P < 0.05), the expression of target protein of Hiwi gene was also significantly inhibited (P < 0.05). The apoptosis rate of MDA-MB-231 cells was significantly higher than that of NC and Blank groups (P<0.05). Conclusion The apoptosis rate of breast cancer cells MDA-MB-231 was significantly increased after siRNA targeting hiwi gene silencing.

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