1.Construction of CRISPR/Cas9 knock-in system for tardigrade damage suppressor gene Dsup and its effect on HEK 293T cell proliferation
Yuting GAO ; Biao ZHANG ; Yali JIA ; Haiyang WANG ; Tao FAN ; Jiafei XI ; Wen YUE ; Quan ZENG ; Junnian ZHOU
Military Medical Sciences 2024;48(8):586-593
Objective To construct HEK 293T cells that express tardigrade Dsup protein fused with green fluorescent protein copGFP in order to study the effect of Dsup protein on proliferation of HEK 293T cells.Methods The CRISPR/Cas9 gene knock-in system was constructed.The target gene fragments of Dsup,copGFP,EF1α and puromycin were amplified by PCR and inserted into pAAVS1-SFFV to construct the fusion vector of Dsup and copGFP,which was known as pAAVS1-SFFV-Dsup-copGFP-EF1α-Puro.pAAVS1-SFFV-Dsup-copGFP-EF1 α-Puro and pAAVS1-CRISPR-Cas9 vector were co-transfected into HEK 293T cells before Dsup gene was inserted into the AAVS1 region of HEK 293T cells via homologous recombination.The HEK 293T cells expressing Dsup gene were obtained following puromycin selection,flow cytometry sorting and genome identification.The expression of Dsup at mRNA and protein levels and proliferation-related genes(MCM2,MCM4,PCNA,Ki-67)were examined to investigate the effects of Dsup gene on the proliferation of HEK 293T-Dsup-copGFP cells.Results The pAAVS1-SFFV-Dsup-copGFP-EF1α-Puro recombinant vector was constructed,and the HEK 293T-Dsup-copGFP cells with Dsup gene inserted in the AAVS1 region were obtained,where both Dsup mRNA and protein were expressed.The cell proliferation rate of HEK 293T-Dsup-copGFP was higher than that of HEK 293T-Control-copGFP(P<0.001).Further investigation revealed that the expressions of Ki-67 and MCM4 protein in HEK 293T-Dsup-copGFP were significantly higher than in the control group,indicating that the knock in of Dsup gene might enhance the proliferation ability of human cells by promoting the expression of Ki-67 and MCM4 protein.Conclusion A gene editing vector is constructed,and stable cell line HEK 293T-Dsup-copGFP for Dsup fusion expression with copGFP is established.The expression of Dsup gene in HEK 293T cells can promote cell proliferation,possibly by upregulating the expressions of Ki-67 and MCM4 protein.
2.Expression of SMOC2 in papillary thyroid carcinoma and its clinicopathological significance
Longyun FAN ; Wei PENG ; Meiling WANG ; Yali ZHAO ; Han XIAO ; Wanxiang WANG ; Qiang WU ; Xian WANG
Chinese Journal of Clinical and Experimental Pathology 2023;39(12):1492-1496,1502
Purpose To investigate the expression of SMOC2 in papillary thyroid carcinomas(PTC)and its efficacy in joint diagnosis with CK19,Galectin-3,MC and BRAF V600E.Methods Bioinformatics was uesd to analyze the mR-NA expression differences of SMOC2 in PTC and benign thyroid tissues in the Gene Expression Omnibus database and The Canc-er Genome Atlas database.Detection of SMOC2 protein expres-sion in paraffin tissue of 75 cases of PTC and 45 cases of papilla-ry thyroid hyperplasia(PTH)was used by using EnVision meth-od,combined with CK19,Galectin-3,and MC and BRAF V600E for sensitivity and specificity analysis.Results The bioinformatics analysis results showed that the mRNA expression level of SMOC2 in PTC tissue was significantly lower than that in benign thyroid tissue(P<0.05),and the area under the curve(AUC)predicted by SMOC2 for PTC diagnosis was 0.910(P<0.001).The immunohistochemical results showed that the ex-pression of SMOC2 in PTC was significantly lower than that in PTH tissue(P<0.001),and the AUC of SMOC2 for PTC diag-nosis was 0.898(P<0.001).The AUC of SMOC2 combined with CK19,Galectin-3,MC and BRAF V600E in the diagnosis of PTC was 1.000(P<0.001),and the AUC values of the combination of other markers were lower than 1.000.Conclu-sion The expression of SMOC2 in PTC is significantly de-creased,which can be used as an important marker for the diag-nosis and differential diagnosis of PTC.Combined with CK19,Galectin-3,MC and BRAF V600E,the sensitivity and specifici-ty of PTC can be improved to a certain extent.
3.Application of continuous renal replacement therapy combined with extracorporeal carbon dioxide removal in children with acute respiratory failure
Wei DANG ; Qin JIANG ; Yali FU ; Yanli ZHANG ; Sufang ZHANG ; Fan ZHANG ; Lei LIU ; Xia LIN
Chinese Journal of Emergency Medicine 2023;32(6):761-767
Objective:To explore the role of continuous renal replacement therapy (CRRT) combined with extracorporeal carbon dioxide removal (ECCO 2R) in the treatment of children with respiratory failure. Methods:The clinical data of 12 children with respiratory failure who were treated with CRRT+ECCO 2R in PICU of Jinan Children's Hospital from July 2020 to August 2022 were collected and analyzed retrospectively. The outcomes and the external pipeline usage of the patients were observed, and the blood gas analysis and ventilator parameters before 1 h and after 1, 6, 12 and 24 h of the treatment were compared by one-way ANOVA with LSD post hoc correction. Results:Six patients successfully withdrew from CRRT+ECCO 2R and mechanical ventilation, three patients were transferred to ECMO treatment. Three cases died after voluntary withdrawal of treatment, and two cases died due to treatment failure. The mortality rate was 41.7%. After continuous treatment of CRRT+ECCO 2R for 15 to 112 h, two cases experienced extracorporeal circuit obstruction. After 1 h of treatment, PaCO 2 decreased from (64.67±24.4) mmHg to (49.42±15.54) mmHg, pH increased from (7.28±0.20) to (7.38±0.11), FiO 2 decreased from (0.85±0.13) to (0.78±0.15), PC decreased from (19.42±4.34) cmH 2O to (17.75±4.00) cmH 2O. After 24 h of treatment, PaCO 2 decreased to (39.2±5.55) mmHg, pH increased to (7.41±0.04), FiO 2 decreased to (0.46±0.11), and PC decreased to (13.8±3.36) cmH 2O, and the differences were statistically significant compared with before treatment ( P < 0.05). Conclusions:The combination of CRRT and ECCO 2R therapy can safely substitute for partial lung ventilation/perfusion function, and play a role in protecting right heart function and improving lung-kidney interaction. It can be considered as an option for extracorporeal respiratory, circulatory, and renal support, and consequently has broad prospects.
4.Analysis of the efficacy of anal dimple anorectoplasty on female infants with congenital anal atresia combined rectal vestibular fistula
Yuhang YUAN ; Xiangyu ZHANG ; Heying YANG ; Xiaoming LIU ; Fan SU ; Ming YUE ; Daokui DING ; Yan′an LI ; Beibei SUN ; Yali JIN
Chinese Journal of Applied Clinical Pediatrics 2023;38(9):698-701
Objective:To investigate the clinical efficacy of anal dimple anorectoplasty on female infants with congenital anal atresia combined rectal vestibular fistula.Methods:Clinical data of 69 female infants with congenital anal atresia combined rectal vestibular fistula admitted to Department of Pediatric Surgery, the First Affiliated Hospital of Zhengzhou University from July 2012 to July 2022 were retrospectively analyzed.They were divided into 2 groups according to the surgical methods: 34 cases of anal dimple anorectoplasty(group A) and 35 cases of anterior sagittal anorectoplasty(group B). The operation time, length of stay, short-term complications, long-term complications and bowel function (determined by the Rintala score at 6 months postoperatively) of the two groups were compared.The difference in the incidence of postoperative complications between groups was compared by Chi- square test, and the remaining differences between groups were compared by the paired t-test. Results:The operative time [(80.18±9.29) min vs.(103.85±8.26) min] and postoperative hospital stay[(6.10±1.52) d vs.(7.63±2.40) d] in group A were significantly shorter than those of group B ( t=11.40, 2.62; all P<0.05). The Rintala total score at 6 months postoperatively in group A was significantly higher than that of group B[(19.36±0.93) points vs.(18.76±0.44) points]( t=3.20, P<0.05). There were no significant differences in the incidences of short-term [(4/34, 11.8%) vs.(7/35, 20.0%)] and long-term complications [(2/34, 5.9%) vs.(4/35, 11.4%)]between group A and group B ( χ2=0.75, 0.75; all P>0.05). Conclusions:Anal dimple anorectoplasty for female infants with congenital anal atresia combined rectal vestibular fistula is safe and effective.
5.Blood eosinophils and clinical features of pneumoconiosis complicated with chronic obstructive pulmonary disease: A cross-sectional study
Yali FAN ; Ruimin MA ; Jingwei WANG ; Xuqin DU ; Qiao YE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(8):605-611
Objective:To analyze the correlation between peripheral blood eosinophil (EOS) level and clinical characteristics of patients with pneumoconiosis complicated with chronic obstructive pulmonary disease (COPD) .Methods:From January 2007 to November 2020, newly diagnosed patients with pneumoconiosis complicated with COPD in Beijing Chaoyang Hospital, were retrospectively analyzed. These patients were stratified into EOS<100 cells/μl group and EOS≥100 cells/μl group, taking 100 cells/μl as the cut-off value. Demographic characteristics, clinical symptoms, lung function and laboratory indexes were compared between the two groups.Results:The median EOS count of patients with pneumoconiosis complicated with COPD was 100 (40, 180) cells/μl. 50.2% (160/319) had blood eosinophil counts ≥100 cells/μl, and 11.0% (35/319) had blood eosinophil counts ≥300 cells/μl. In comparison with EOS<100 cells/μl group, EOS≥100 cells/μl group were older ( P=0.035), had higher body mass index ( P=0.008), and had lower forced respiratory volume in the first second ( P=0.017), had higher the ratio of residual volume to total lung volume ( P=0.010), and had lower diffusing capacity of the lung for carbon monoxide ( P=0.008). Arterial partial pressure of oxygen was significantly reduced in EOS≥100 cells/μl group ( P=0.039). The peripheral blood EOS count was negatively correlated with forced vital capacity, forced breathing volume in the first second, carbon monoxide diffusion, peak expiratory flow, and maximum mid expiratory flow as a percentage of expected values ( rs=-0.22, -0.18, -0.19, -0.19, -0.19, P=0.000, 0.001, 0.003, 0.008, 0.002), and positively correlated with the ratio of residual air volume to total lung volume ( rs=0.17, P=0.002) . Conclusion:There was a correlation between blood EOS count and pulmonary function parameters, can proide reference for the diagnosis and treatment of chnoric obstuctive pulmmory disease in clinical practice.
6.Nam Dinh virus was detected and isolated in arbovirus investigation in Shanxi, China
Shenghui LIU ; Xiaodong TIAN ; Weijia ZHANG ; Hongmei ZHENG ; Junying ZHAO ; Chongxiao XU ; Yali ZHANG ; Shihong FU ; Kai NIE ; Fan LI ; Peifang DAI ; Qikai YIN ; Ying HE ; Jingxia CHENG ; Huanyu WANG
Chinese Journal of Experimental and Clinical Virology 2023;37(1):25-29
Objective:In this study, the collected mosquito samples were subjected to viral isolation to identify the species and branch characteristics of arboviruses in five regions of Shanxi Province.Methods:Eight arboviruses in mosquito samples collected from July to September 2020 were detected by real-time fluorescent quantitative PCR, and virus isolation was carried out through cell culture. Virus isolates were identified and analyzed by molecular biology and bioinformatics method.Results:We detected 1 batch of positive samples of Japanese encephalitis virus, 2 batches of positive samples of Culex flavivirus and 8 batches of positive samples of Nam Dinh virus among 121 batches of mosquito samples. Seven virus isolates were isolated, numbered: SX-YJ-Cxp-4、SX-YJ-Ars-2、SX-YJ-Cxp-1、SX-LY-Cxp-10、SX-GP-Ars-5、SX-GP-Cxp-2、SX-GP-Cxp-4, all of which were identified as Nam Dinh virus, and the whole genome sequencing was performed on one of them, and the result showed that Shanxi Nam Dinh virus isolate and Yunnan Nam Dinh virus isolate belonged to the same evolutionary branch.Conclusions:Nam Dinh virus was isolated and identified on the specimen from Shanxi province for the first time.
7.Blood eosinophils and clinical features of pneumoconiosis complicated with chronic obstructive pulmonary disease: A cross-sectional study
Yali FAN ; Ruimin MA ; Jingwei WANG ; Xuqin DU ; Qiao YE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(8):605-611
Objective:To analyze the correlation between peripheral blood eosinophil (EOS) level and clinical characteristics of patients with pneumoconiosis complicated with chronic obstructive pulmonary disease (COPD) .Methods:From January 2007 to November 2020, newly diagnosed patients with pneumoconiosis complicated with COPD in Beijing Chaoyang Hospital, were retrospectively analyzed. These patients were stratified into EOS<100 cells/μl group and EOS≥100 cells/μl group, taking 100 cells/μl as the cut-off value. Demographic characteristics, clinical symptoms, lung function and laboratory indexes were compared between the two groups.Results:The median EOS count of patients with pneumoconiosis complicated with COPD was 100 (40, 180) cells/μl. 50.2% (160/319) had blood eosinophil counts ≥100 cells/μl, and 11.0% (35/319) had blood eosinophil counts ≥300 cells/μl. In comparison with EOS<100 cells/μl group, EOS≥100 cells/μl group were older ( P=0.035), had higher body mass index ( P=0.008), and had lower forced respiratory volume in the first second ( P=0.017), had higher the ratio of residual volume to total lung volume ( P=0.010), and had lower diffusing capacity of the lung for carbon monoxide ( P=0.008). Arterial partial pressure of oxygen was significantly reduced in EOS≥100 cells/μl group ( P=0.039). The peripheral blood EOS count was negatively correlated with forced vital capacity, forced breathing volume in the first second, carbon monoxide diffusion, peak expiratory flow, and maximum mid expiratory flow as a percentage of expected values ( rs=-0.22, -0.18, -0.19, -0.19, -0.19, P=0.000, 0.001, 0.003, 0.008, 0.002), and positively correlated with the ratio of residual air volume to total lung volume ( rs=0.17, P=0.002) . Conclusion:There was a correlation between blood EOS count and pulmonary function parameters, can proide reference for the diagnosis and treatment of chnoric obstuctive pulmmory disease in clinical practice.
8.Summary of the best evidence for the evaluation and management strategy of Non -suicidal self -injury in adolescents
Yali HU ; Linbo LI ; Xiaofen FAN ; Yan LI ; Guizhi QIAO ; Qian ZHANG
Chinese Journal of Modern Nursing 2023;29(17):2260-2267
Objective:To summarize the best evidence for the evaluation and management strategies of adolescent non-suicidal self-injury, and provide a basis for medical staff to carry out standardized evaluation and management of adolescent non suicidal self injury.Methods:According to evidence-based nursing methods and the "6S"evidence pyramid model, the evidences of non-suicidal self-injury in adolescents, including guidelines, standards, case report, evidence summary, randomized controlled trial (RCT) , systematic review, clinical practice and other evidences were searched in BMJ Best Practice, UpToDate, World Health Organization, National Institute for Health and Care Excellence, National Guidelines Clearinghouse, Registered Nurses ' Association of Ontario, Scottish Intercollegiate Guidelines Network, American College of Physicians Club, Australian Joanna Briggs Institute (JBI) Evidence-based Health Care Center Library, Cochrane Library, PubMed, Embase, PsycINFO, China Biology Medicine disc, China National Knowledge Infrastructure, Wanfang Database, Medlive and other databases. The retrieval period was from the establishment of the databases to June 30, 2022. The quality evaluation of the articles was evaluated by two researchers trained in the evidence-based curriculum. Included studies were pre-graded using the Australian JBI Evidence-based Health Centre Evidence pre-grading system and the Evidence Recommendation Level System (2014 edition) . Results:Finally, a total of 18 articles were included, including 5 guidelines, 2 standards, 1 case report, 1 evidence summary, 3 systematic reviews, 1 Meta-analysis, 2 clinical practices and 3 RCT studies. A total of 22 pieces of best evidence were summarized from 6 dimensions, such as screening and assessment, multi-professional and multi-form intervention strategies, training and education, self-management strategies, disposal after self-injury treatment and follow-up.Conclusions:The best evidence based on evidence-based assessment and management strategies of adolescent non-suicidal self-injury is scientific and practical, which can provide reference for the clinical practice of adolescent non-suicidal self-injury.
9.Analysis of the status quo and influencing factors of social alienation in young adults undergoing radioactive iodine therapy for papillary thyroid carcinoma
Yongbo WANG ; Yonglin WANG ; Peixia WANG ; Yali CHEN ; Yuxia FAN
Chinese Journal of Modern Nursing 2023;29(32):4404-4408
Objective:To understand the status quo of social alienation in young adults undergoing radioactive iodine (RAI) therapy for papillary thyroid carcinoma (PTC) , analyze its influencing factors, and provide references for the formulation of clinical nursing interventions.Methods:Totally 206 young adults with PTC who underwent RAI therapy at the First Affiliated Hospital of Zhengzhou University from January 2021 to May 2022 were selected by convenience sampling. These patients were surveyed using a general information questionnaire, General Alienation Scale (GAS) , Social Support Rating Scale (SSRS) , and Hospital Anxiety and Depression Scale (HADS) . Multiple linear regression analysis was utilized to analyze the influencing factors of social alienation in young adults undergoing RAI therapy for PTC.Results:The score of social alienation in the young adults undergoing RAI therapy for PTC was (39.39±4.93) . Univariate analysis results indicated that there was a statistically significant difference in the scores of social alienation among the young PTC patients with different marital statuses, levels of disease awareness, employment statuses, family per capita monthly incomes, and tumor stages ( P<0.05) . Multiple linear regression analysis revealed that the influencing factors of social alienation included the level of disease awareness, family per capita monthly income, tumor staging, social support score, and anxiety and depression score ( P<0.05) . Conclusions:The overall level of social alienation in young adults undergoing RAI therapy for PTC is moderate. Healthcare providers should timely assess and adopt targeted intervention measures to enhance patients' sense of social participation.
10.Effects of needle-embedding therapy on improvement of pneumoperitoneum and abdominal distension after colonoscopy
Yali ZHANG ; Ying ZHANG ; Meiqing FAN
Chinese Journal of Primary Medicine and Pharmacy 2022;29(5):651-654
Objective:To investigate the effects of needle-embedding therapy on improvement of pneumoperitoneum and abdominal distension after colonoscopy.Methods:A total of 102 patients who received colonoscopy in Hangzhou Hospital of Traditional Chinese Medicine, China between June 2018 and June 2020 were included in this study. They were randomly assigned to observation and control groups ( n = 51 per group). The control group was subjected to routine symptomatic treatment and the observation group was given needle-embedding therapy. The improvement effects on pneumoperitoneum and abdominal distension, examination duration, numerical rating scale score, and grade of abdominal distension at 6 and 24 hours after colonoscopy were compared between observation and control groups. Results:Total response rate in the observation group was significantly higher than that in the control group [92.16% (47/51) vs. 74.51% (38 /51), χ 2 = 5.71, P < 0.05]. The time to enteroscope entry and the time to enteroscope withdrawal were (4.86 ± 1.23) minutes and (5.24 ± 1.13) minutes, respectively, which were significantly shorter than those in the control group [(8.79 ± 1.25) minutes, (6.97 ± 1.12) minutes, t = 16.00, 7.76, both P < 0.001]. Numerical rating scale score in the observation group was significantly lower than that in the control group [(1.50 ± 0.41) points vs. (2.30 ± 0.42) points, t = 9.73, P < 0.001). At 6 and 24 hours after colonoscopy, the amplitude of improvement of abdominal distension in the observation group was significantly greater than that in the control group (both P < 0.05). Conclusion:Needle-embedding therapy can effectively improve pneumoperitoneum and abdominal distension after enteroscopy, shorten the examination duration, and relieve pain.

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