1.Stable knockout of ACSS3 in lung cancer cell line using CRISPR/Cas 9 technology
Qianqian HUANG ; Yufang JIA ; Huajun YU ; Rongrong CHEN ; Lili CHEN ; Jun WU ; Haitao ZHANG
Basic & Clinical Medicine 2025;45(8):1016-1021
Objective To explore the effect of acyl-CoA synthetase short-chain family member 3(ACSS3)gene on the proliferation of human large cell lung cancer cells(NCI-H460)using CRISPR/Cas 9 gene editing technology.Methods The expression of ACSS3 was detected by Western blot.ACSS3-targeting sgRNAs were designed,and a CRISPR/Cas 9 knockout vector was constructed and transfected into NCI-H460 cells.The transfected cells were selected with puromycin based on vector-carried resistance.ACSS3-knockout monoclonal cell strains were established by limited dilution method and then expanded in culture.Knockout efficiency was confirmed by Western blot.Cell proliferation was assessed using MTT and colony formation assays.Results The expression of ACSS3 was significantly elevated in NCI-H460 cells as compared with human normal lung epithelial cells BEAS-2B(P<0.05).No ACSS3 protein was detected in the knockout monoclonal strain,indicating successful generation of ACSS3-knockout NCI-H460 cells.Compared with the control cells transfected with empty vector,the proliferation and colo-ny formation ability were inhibited in NCI-H460 cells with ACSS3 knockout(P<0.05).Conclusions The ACSS3-knockout NCI-H460 cell strain was successfully established,which provides a foundation for further study on the role of ACSS3 in lung cancer.
2.Effects of grain-sized moxibustion on Th1/Th2 balance and transcription factors T-bet and GATA3 in immunosuppressed mice induced by cyclophosphamide
Tao ZHU ; Zhenzhi WANG ; Jia REN ; Yanting CHENG ; Zhen GAO ; Yufang JI ; Jinling MIAO ; Laixi JI
Journal of Beijing University of Traditional Chinese Medicine 2024;47(6):818-825
Objective To observe the effects of grain-sized moxibustion on Th1 cell/Th2 cell(Th1/Th2)balance and transcription factors T-box transcription factor(T-bet)and GATA binding protein 3(GATA3)in immunosuppressive mice induced by chemotherapy.Methods According to the random number table method,32 SPF male CD-1(ICR)mice were randomly divided into the normal group,model group,levamisole hydrochloride group,and grain-sized moxibustion group,with eight mice per group.Except for the normal group,the immunosuppressive model was established by intraperitoneal injection of cyclophosphamide(80 mg/kg,once daily for three consecutive days).Mice in the levamisole hydrochloride group were given levamisole hydrochloride solution(10 mg/kg)by gavage.Mice in the grain-sized moxibustion group was given grain-sized moxibustion at"Guanyuan"(CV4),bilateral"Zusanli"(ST36),and bilateral"Sanyinjiao"(SP6),with three Zhuang at each acupoint for approximately 30 s each.The intervention was administered once daily for seven consecutive days.The general condition of mice was observed.The spleen mass and spleen index were detected.The pathological changes of spleen tissue were observed by HE staining.The protein and mRNA expressions of T-bet,GATA3,interferon-γ(IFN-γ),and interleukin(IL)-4 in spleen tissue of mice were detected by Western blotting and real-time PCR.The contents of IFN-γ,IL-2,and IL-4 in serum of mice were detected by enzyme-linked immunosorbent assay.Results Compared with the normal group,the mice in the model group were slow in response,unstable in gait;the spleen weight and spleen index were increased(P<0.05);the structure of spleen tissue was disordered,the mRNA and protein expressions of T-bet and IFN-γ in spleen tissue were decreased,and the mRNA and protein expressions of GATA3 and IL-4 were increased(P<0.05);the contents of IFN-γ and IL-2 in serum were decreased,and the content of IL-4 was increased(P<0.05).Compared with the model group,the general condition of mice in the levamisole hydrochloride group and the grain-sized moxibustion group was improved,the structure of spleen tissue was improved,the mRNA and protein expressions of T-bet and IFN-γ in spleen tissue were decreased,and the mRNA and protein expressions of GATA3 and IL-4 were increased(P<0.05);the contents of IFN-γ and IL-2 in serum were decreased,and the content of IL-4 was increased(P<0.05).Conclusion Grain-sized moxibustion can significantly improve the immunosuppressive symptoms induced by chemotherapy.The mechanism may be through regulating the expressions of transcription factors T-bet and GATA3,regulating Th1/Th2 balance,and thus restoring the immune balance.
3.Development of a working model of evidence-based nursing practice in deep vein thrombosis prophylaxis
Yu WANY ; Yufang HAO ; Yufen MA ; Yuan XU ; Ranxun AN ; Haibo DENG ; Lei WANG ; Xiaojie WANG ; Jianhua SUN ; Jia LIU ; Liyun ZHU ; Xinjuan WU
Chinese Journal of Nursing 2024;59(15):1804-1811
Objective To construct an evidence-based practice model for nurses in preventing deep vein thrombosis(DVT)and provide a scientific and targeted theoretical basis for nurses to carry out evidence-based nursing practice in DVT prevention.Methods Based on the previous evidence-based nursing practice project on DVT prevention after hip and knee arthroplasty,the research team used theoretical analysis and brainstorming to develop a draft of the work model.Expert meetings were organized to validate the content of the draft using the Delphi method,leading to the finalization of the evidence-based practice model for nurses in preventing DVT.Results The Knowledge-to-Action(KTA)framework was selected as the basic framework for constructing the evidence-based nursing practice model for preventing DVT.Theoretical Domain Framework,Theory of Planned Behavior,and Social Cognitive Theory were chosen to explore the influencing factors of nurses'behavior change in preventing DVT through evidence-based practice.The authority coefficient of the participating experts was 0.904,indicating high reliability.The final model consisted of 6 key components:knowledge generation,problem identification,localization and adaptation,knowledge application,sustained knowledge use,and conceptual framework for behavior change through evidence-based practice.Conclusion Based on theoretical analysis and clinical practice,this study developed an evidence-based practice model for nurses in preventing DVT using the expert meeting.The research methodology was scientific,and the content was reliable,providing a theoretical basis for nurses to engage in evidence-based nursing practice for DVT prevention.
4.Meta-analysis of implicit relational assessment procedure in measuring implicit body image bias among young and middle-aged people
Junying FAN ; Yannan JIA ; Li LI ; Yufang GUO
Chinese Journal of Modern Nursing 2024;30(27):3700-3707
Objective:To explore the implicit bias of body image measurement in young and middle-aged people using implicit relational assessment procedure (IRAP) .Methods:ProQuest, Scopus, EBSCO, Cochrane Library, Web of Science, PubMed, China National Knowledge Infrastructure, WanFang data, and VIP databases were systematically searched to collect articles on the use of IRAP to measure implicit bias in body image in young and middle-aged populations. The retrieval period was from the establishment of the databases to December 15, 2023. Meta-analysis was conducted using Review Manager 5.4 software.Results:A total of 11 articles were included. The results of the Meta-analysis showed that the score of D general (reaction time index of pro-thinness and anti-fat) was 0.09 (95% CI: 0.07~0.11, P<0.01), indicating that compared with overweight/obesity, young and middle-aged people had a strong sense of identification with thinness and a stronger desire for thinness. The score of D thin (reaction time index of pro-thinness) was 0.26 (95% CI: 0.12~0.39, P<0.01), indicating that young and middle-aged people' reaction speed was faster when they agreed with thinness and participants had the desire to be thin. The score of D fat (reaction time index of anti-obesity) was -0.09 (95% CI: -0.21~0.03, P>0.05), indicating that young and middle-aged people had no bias against overweight/obesity. Conclusions:Young and middle-aged people generally have the performance of implicit thinness, with a strong desire for thinness and no implicit bias towards overweight.
5.Role of STAT3/NCOA4-mediated ferritinophagy in salidroside-induced attenuation of intestinal ischemia-reperfusion injury in mice: relationship with ferroptosis
Xinrun WANG ; Yufang LENG ; Huanhuan GAO ; Xiaojing JIA
Chinese Journal of Anesthesiology 2023;43(11):1316-1321
Objective:To evaluate the role of signal transducer and activator of transcription 3 (STAT3)/nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in salidroside-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty-six SPF-grade healthy male C57BL mice, aged 6-8 weeks, weighing 20-25 g, were divided into 6 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation+ salidroside group (SS group), intestinal I/R group (IR group), intestinal I/R+ salidroside group (IS group), intestinal I/R+ salidroside+ autophagy activator rapamycin group (ISR group) and intestinal I/R+ salidroside+ STAT3 activator colivelin group (ISC group). The intestinal I/R injury model was established by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR, IS, ISR and ISC groups, while the superior mesenteric artery was only isolated without clipping in S and SS groups. At 1 week before developing the model, salidroside 40 mg/kg was intraperitoneally injected once a day for 7 consecutive days in SS, IS, ISC and ISR groups, rapamycin 4 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISR, colivelin 1 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISC, while the equal volume of normal saline was given instead in the rest two groups. The mice were sacrificed at 30 min of reperfusion, and intestinal tissues were obtained for examination of the pathological changes after HE staining (with a optical microscope) which were scored according to Chiu and for determination of contents of malondialdehyde (MDA), Fe 2+, glutathione (GSH) and reactive oxygen species (ROS), activity of superoxide dismutase (SOD) and expression of p-STAT3, STAT3, glutathione peroxidase 4 (GPX4), NCOA4 and ferritin heavy chain 1 (FTH1) in intestinal tissues (by Western blot). Results:Compared with group S, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, the content of GSH was decreased, the activity of SOD was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, the p-STAT3/STAT3 ratio was increased ( P<0.05), pathological injury was found in intestinal tissues, and no significant change was found in the aforementioned indexes in group IR( P>0.05). Compared with group IR, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly decreased, GSH content was increased, SOD activity was increased, the expression of p-STAT3 and NCOA4 was down-regulated, the expression of GPX4 and FTH1 was up-regulated, p-STAT3/STAT3 ratio was decreased ( P<0.05), and the pathological injury was significantly alleviated in intestinal tissues in group IS. Compared with group IS, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, GSH content was decreased, SOD activity was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, p-STAT3/STAT3 ratio was increased ( P<0.05), and the pathological injury was aggravated in intestinal tissues in ISR and ISC groups. There was no statistically significant difference in the expression of STAT3 among the five groups ( P>0.05). Conclusions:STAT3/NCOA4-mediated ferritinophagy is involved in the process of salidroside-induced reduction of intestinal I/R injury in mice, which may be related to inhibiting ferroptosis.
6.Role of endoplasmic reticulum stress-mediated apoptosis in proanthocyanidins-induced attenuation of intestinal ischemia-reperfusion injury in mice
Xiaojing JIA ; Yufang LENG ; Xinrun WANG ; Huanhuan GAO
Chinese Journal of Anesthesiology 2023;43(12):1439-1444
Objective:To evaluate the role of endoplasmic reticulum stress-mediated apoptosis in proanthocyanidins-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy adult male C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 5 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation + proanthocyanidins group (S+ PC group), intestinal I/R group (I/R group), intestinal I/R + proanthocyanidins group (I/R+ PC group) and intestinal I/R + proanthocyanidins + tunicamycin group (I/R+ PC+ TM group). The superior mesenteric artery was clamped for 60 min and reperfused for 120 min to establish a mouse intestinal I/R injury model.Proanthocyanidin 100 mg/kg was given by intragastric gavage every day 1 week before ischemia in S+ PC group, I/R+ PC group and I/R+ PC+ TM group, and the equal volume of normal saline was given for 7 consecutive days in S group and I/R group, and endoplasmic reticulum stress agonist tunicamycin 1 mg/kg was intraperitoneally injected at 24 h before ischemia in I/R+ PC+ TM group.The mice were sacrificed at 120 min of reperfusion, and the small intestinal tissues were taken for microscopic examination of the histopathological changes (using light microscopy) and for determination of the level of diamine oxidase (DAO) (by enzyme-linked immunosorbent assay), cell apoptosis (by TUNEL method), glucose regulatory protein 78 (GRP78), C/EBP-homologous protein (CHOP) and cleaved caspase-3, Bax and Bcl-2 (by Western blot). Intestinal damage was assessed and scored according to Chiu, and the apoptosis index (AI) and Bcl-2/Bax ratio were calculated. Results:Compared with S group, the Chiu′s score, level of DAO and AI were significantly increased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, the ratio of Bcl-2/Bax was decreased( P<0.05), and the pathological damage occurred in the small intestinal tissue in I/R group, and no significant change was found in the aforementioned indexes in S+ PC group ( P>0.05). Compared with I/R group, the Chiu′s score, DAO level and AI were significantly decreased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was down-regulated, the expression of Bcl-2 was up-regulated, the ratio of Bcl-2/Bax was increased( P<0.05), and the pathological injury to the small intestinal tissue was significantly reduced in I/R+ PC group. Compared with I/R+ PC group, the Chiu′s score, level of DAO and AI were significantly increased, the expression of GRP78, CHOP, cleaved caspase-3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, and the ratio of Bcl-2/Bax was decreased( P<0.05), and the pathological damage to the small intestinal tissue was aggravated in I/R+ PC+ TM group. Conclusions:Proanthocyanidins can alleviate intestinal I/R injury by inhibiting endoplasmic reticulum stress-mediated cell apoptosis in mice.
7.Role of SIRT1/Nrf2 signaling pathway in berberine preconditioning-induced reduction of intestinal ischemia-reperfusion injury in mice and the relationship with ferroptosis
Huanhuan GAO ; Yufang LENG ; Xinrun WANG ; Xiaojing JIA
Chinese Journal of Anesthesiology 2023;43(12):1515-1519
Objective:To evaluate the role of silencing information regulatory factor 1/nuclear factor E2 related factor 2 (SIRT1/Nrf2) signaling pathway in berberine preconditioning-induced reduction of intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty-six SPF-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 22-25 g, were divided into 6 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation + berberine preconditioning group (SB group), intestinal I/R group (IR group), intestinal I/R + berberine preconditioning group (B group), intestinal I/R + berberine preconditioning + SIRT1 inhibitor EX527 group (BE group) and berberine preconditioning + intestinal I/R + ferroptosis inducer RSL3 group (BR group). The model of intestinal I/R injury was prepared by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR group, B group, BE group and BR group, while the superior mesenteric artery was only isolated without ligation in S group and SB group. Berberine 50 mg/kg was administered by intragastric gavage once a day starting from 7 days before developing the model in SB group, B group, BE group and BR group. EX527 5 mg/kg and RSL3 5 mg/kg were intraperitoneally injected once a day at 3 days before surgery in BE group and BR group, respectively. The equal volume of normal saline was given in the other groups. The mice were sacrificed at 30 min of reperfusion, and the intestinal tissues were taken for microscopic examination of the pathological changes of intestinal mucosa (with a light microscope) which was scored according to Chiu and for determination of the contents of Fe 2+ and malondialdehyde (MDA) and superoxide dismutase (SOD) activity (by colorimetry), glutathione (GSH) content (by enzyme-linked immunosorbent assay), reactive oxygen species (ROS) content (by fluorescence staining), and expression of glutathione peroxidase 4 (GPX4), SIRT1 and Nrf2 (by Western blot). Results:Compared with S group, Chiu′s score was significantly increased, the contents of Fe 2+, MDA and ROS were increased, the content of GSH and activity of SOD were decreased, the expression of GPX4 was down-regulated, and the expression of SIRT1 and Nrf2 was up-regulated in IR group ( P<0.05), and no significant change was found in Chiu′s score in SB group ( P>0.05). Compared with IR group, Chiu′s score was significantly decreased, the contents of Fe 2+, MDA and ROS were decreased, the content of GSH and activity of SOD were increased, the expression of GPX4 was up-regulated, and the expression of SIRT1 and Nrf2 was up-regulated in B group ( P<0.05). Compared with B group, Chiu′s score was significantly increased, the contents of Fe 2+, MDA and ROS were increased, the content of GSH and activity of SOD were decreased, and the expression of GPX4 was down-regulated in BE and BR groups, and the expression of SIRT1 and Nrf2 was down-regulated in BE group( P<0.05). Conclusions:The mechanism by which berberine preconditioning reduces intestinal I/R injury may be associated with activation of SIRT1/Nrf2 signaling pathway, thus inhibiting ferroptosis in mice.
8.Association of deficits in prepotent response inhibition with attention deficit and impulsive behavior among children with attention deficit hyperactivity disorder
HE Jia ; XU Fangzhong ; TANG Luhan ; YU Ge ; HAN Yufang
Journal of Preventive Medicine 2023;35(9):766-769
Objective:
To investigate the associations of deficits in prepotent response inhibition with attention deficit and impulsive behavior in children with attention deficit hyperactivity disorder (ADHD), so as to provide insights into response inhibition training in ADHD to relieve clinical symptoms.
Methods:
From March to December 2022, 57 children with ADHD were selected from the clinical psychology department of a tertiary hospital in Hangzhou City as the ADHD group, and 55 normal children matched by age and gender were selected from a primary school as the control group. Prepotent response inhibition, attention deficit and impulsive behavior were assessed by inhibition conflict task, visual continuous performance test (CPT) and matching familiar figures test (MFFT), respectively. The associations of deficits in prepotent response inhibition with attention deficit and impulsive behavior were analyzed using multiple linear regression model, and the predictive value of deficits in prepotent response inhibition on ADHD was evaluated using receiver operating characteristic (ROC) curve.
Results:
The children included in the ADHD group had a mean age of (8.77±1.60) years and 44 males (77.19%), and the children included in the control group had a mean age of (9.20±1.77) years and 42 males (76.36%). The error rate of inhibition conflict task, missing report rate of visual CPT and the number of MFFT errors were higher in the ADHD group than in the control group [22.50% (12.50%) vs. 8.75% (7.00%), 24.00% (30.00%) vs. 7.50% (7.00%), 8.67±3.32 vs. 4.47±3.16; all P<0.05]. Multiple linear regression model showed that the error rate of inhibition conflict task was positively associated with the missing report rate of visual CPT (R2=0.135, β=0.091, P<0.001) and the number of MFFT errors (R2=0.092, β=0.009, P<0.001). The area under the ROC curve was 0.891, the sensitivity was 93.00%, the specificity was 80.00%, and the cut-off was 13.13%.
Conclusions
The deficits in prepotent response inhibition are positively associated with attention deficit and impulsive behavior.
9.Status quo of nursing for elderly functional constipation based on clinical practice guidelines
Danjing ZHANG ; Ying HE ; Jia YAO ; Weizi WU ; Meiyan FAN ; Yufang HAO
Chinese Journal of Modern Nursing 2022;28(21):2878-2883
Objective:To explore the gap between the clinical nursing status and practice guidelines of elderly functional constipation, propose corresponding action strategies, and provide reference for the clinical transformation of the guidelines.Methods:From July to October 2019, totally 330 clinical nurses and nursing managers of elderly patients with functional constipation admitted into the general hospitals, hospitals of traditional Chinese medicine, and hospitals of traditional Chinese medicine and western medicine in Beijing were selected by convenient sampling and investigated with self-made questionnaires. The standard and algorithm of recommendations were formulated, and Excel software was used to calculate the compliance rate, partial compliance rate, and non-compliant rate of recommendations. A total of 330 questionnaires were distributed in this study, and 330 questionnaires were recovered, with a recovery rate of 100.0% (330/330) . After excluding invalid questionnaires such as items with missing values greater than 10% and all items having the same answers, there were 317 valid questionnaires, with an effective rate of 96.1% (317/330) .Results:Among the recommendations of the 12 guidelines, the 3 items with the highest compliance rate were health education related to toileting patterns, health education related to activities and sports, satisfaction and effect evaluation, while the 4 items with the highest non-compliance rates were building interdisciplinary teams, comprehensive education and program promotion, health assessment, and physical examination of medication history. There was a statistically significant difference in the clinical practice status between nurses who had and had not received relevant training ( P<0.01) . Conclusions:Most nurses can provide comprehensive care for elderly patients with functional constipation, but there is a big gap between the practice and the guidelines. Measures such as strengthening nurse training and talent training and standardizing the nursing process of constipation can help promote the clinical translation of evidence.
10.Research study on nucleic acid testing of hemostatic stickers used and the surface of the recipients′ hands after the inoculation of inactivated SARS-CoV-2 vaccine
Gaowei FAN ; Yufang LIANG ; Xue MA ; Bing LIU ; Jing WANG ; Lu LU ; Guiqin LI ; Wen JIN ; Jia GU ; Minggang WANG ; Qingtao WANG
Chinese Journal of Preventive Medicine 2021;55(3):406-408
Among the staff of Beijing Chao-Yang Hospital, Capital Medical University, who received the inactivated SARS-CoV-2 vaccine on January 30 in 2021, 28 recipients were selected for this research. Samples for nucleic acid tests were collected from the surface of the recipients′ both hands before and after vaccination. The hemostatic stickers used after the inoculation were also collected for nucleic acid tests. The nucleic acid tests of the samples collected from the surface of both hands of the 28 recipients before vaccination were all negative. After vaccination, the nucleic acid tests of the samples collected from the surface of both hands of recipients were positive in 3 cases, and suspicious in 8 cases, with a positive rate of 10.7%. A total of 25 hemostatic stickers used were collected, 24 of them had positive nucleic acid tests, and the rest one had suspicious nucleic acid test result, with a positive rate of 96%. The hemostatic stickers used after the inoculation have the risk of nucleic acid contamination.


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