1.Construction of anticipatory grief intervention for family caregivers of advanced cancer patients based on narrative theory
Yuzhe JIA ; Xiaolin YANG ; Shuting HUANG ; Zibo FAN ; Lu ZHU ; Di YANG ; Huihui CHEN ; Guangqing ZHANG
Chinese Journal of Practical Nursing 2024;40(11):823-830
Objective:To construct an anticipatory grief intervention scheme for family caregivers of advanced cancer patients based on narrative theory, and to provide reference for anticipatory grief nursing intervention.Methods:From October 2022 to May 2023, through literature research, semi-structured interview and brainstorming method, the first draft of nursing intervention plan was constructed, the Delphi method was used to conduct 2 rounds of correspondence consultation with 15 experts, and the indicators at all levels were modified according to the opinions of experts, and the final draft of intervention plan was formed.Results:The experts were all female, aged (49.67 ± 5.83) years old. The authority coefficient of the two rounds of experts was 0.87. The Kendall coordination coefficients of the first, second, and third level indicators after the first round of expert inquiry were 0.195, 0.113, and 0.093, respectively. The Kendall coordination coefficients of the first, second, and third level indicators after the second round of expert inquiry were 0.200, 0.119, and 0.101, respectively. The differences were statistically significant ( χ2 values were 8.76-107.21, all P<0.05).Finally, a nursing intervention plan based on narrative theory was formed, which included 4 primary indicators, 19 secondary indicators and 72 tertiary indicators. Conclusions:The anticipatory grief intervention scheme for family caregivers of advanced cancer patients is scientific, practical and feasible, and can be used for psychological nursing of family caregivers.
2.Construction and gene identification of myeloid-specific Spi1 knockout mice
Xuming WU ; Huihui WANG ; Xiangling ZHU ; Yuanyuan ZHOU ; Anqi WANG ; Huiru ZHANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(3):413-417
Objective To construct myeloid-specific Spi1 gene knockout mice and analyze their genotypes,so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets.Methods Ac-cording to the principle of CRISPR/Cas9 technology and Cre/LoxP system,sgRNA and Donor vectors were de-signed and constructed.The transcript of Exon 2(Exon 2)was used as the knockout region,and Loxp elements were placed on both sides of Exon 2.Cas9 protein,sgRNA and Donor vector were mixed and microinjected into the fertilized eggs of C57BL/6J mice,the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice,and F0 generation was obtained after 19~20 days.Positive F0 mice were mated with C57BL/6J mice to ob-tain stable F1 Spi1flox/+mice.Spi1flox/+mice of F1 generation were selfed to obtain Spi1flox/flox mice.Spi1flox/flox mated with Lyz2-Cre+mice to obtain Spi1flox/+/Lyz2-Cre+mice,and then mated with Spi1flox/flox,the Spi1flox/flox/Lyz2-Cre+mice were myeloid-specific Spi1 gene knockout(KO)mice.Spi1flox/flox/Lyz2-cre-mice were used as wild-type(WT)mice.DNA of WT and KO mice was extracted,and the genotypes were identified by agarose gel electro-phoresis after PCR amplification.Western blot was used to detect the expression of spleen focus forming virus provi-ral integration oncogene,Spi-1/purine rich box-1(PU.1)in immune cells of WT and KO mice.Results The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1flox/flox homozygote,and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre+.Western blot showed that compared with WT group,the protein PU.1 was not expressed in bone marrow-derived macropha-ges(BMDMs)and peritoneal macrophages(PM)in KO group(P<0.01).There was no significant difference of statistics in the expression level of PU.1 in T cells between KO mice and WT mice.The results of PCR and West-ern blot showed that myeloid-specific Spi1 KO mice were successfully constructed.Conclusion The myeloid-spe-cific Spi1 gene KO mice are successfully constructed and identified,which provides animal model basis for further revealing the potential mechanism of PU.1 inimmune regulation.
3.Breeding and genotyping of T lymphocyte-conditional Spi1 knockout mice
Huihui WANG ; Xiangling ZHU ; Xuming WU ; Huiru ZHANG ; Yuanyuan ZHOU ; Anqi WANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(4):595-599
Objective To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in-vestgation of the specific role of Spi1-encoded protein PU.1.Methods The Lck-Cre mice were mated with Spi1flox/flox mice to obtain Lck-Cre×Spi1flox/flox mice(T lymphocyte-specific Spi1 knockout mice),and the genotype was determined by polymerase chain reaction(PCR)and agarose gel electrophoresis.Magnetic beads were used to sort out the splenic T lymphocytes,and the knockdown efficiency of PU.1 in T cells was detected by Western blot,quantitative real-time PCR(qPCR)and flow cytometry.Results The Lck-Cre×Spi1flox/flox mouse genotype was stably inherited.Compared with Spi1flox/flox mice,the expression level of PU.1 was significantly reduced in splenic T cells of Lck-Cre×Spi1flox/flox mice.Conclusion In this study,the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology,which provided a reliable an-imal model for the subsequent experiments of the specific role of PU.1 in T cell-related diseases.
4.Construction and efficiency detection of Csf1r-CreERT2 R26REYFP reporter gene mouse based on Cre/Loxp system
Xiangling ZHU ; Xuming WU ; Huihui WANG ; Yuanyuan ZHOU ; Anqi WANG ; Huiru ZHANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(7):1175-1180
Objective To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r-CreERT2-mediated enhancement of CSF1R in CD45+cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice,and Csf1r-CreERT2R26REYFP mice were identified through PCR and Western Blot analyses.Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45+cells across different mouse tissues following tamoxifen induction.Results Csf1r-CreERT2 R26REYFP reporter gene mice were acquired.In addition,it was found that Csf1r-CreERT2-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45+cells in different locations.Compared to the R26REYF P group,the highest labeling efficiency was observed in the brain tissue(P<0.001),the lowest in the thymus tissue(P<0.05),and no sig-nificant difference was observed in the spleen tissue.Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice.Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45+cells in different parts of mice.
5.Predictive value of PCT/PLT and CRP/ALB in severe acute pancreatitis and associated liver injury
Mengwei CUI ; Qianqian HE ; Haifeng WANG ; Huihui LI ; Jiye LI ; Zongchao CUI ; Qiaofang WANG ; Sanyang CHEN ; Changju ZHU
Chinese Journal of Emergency Medicine 2024;33(10):1369-1375
Objective:To investigate the predictive value of procalcitonin to platelet ratio (PPR) and C-reactive protein to albumin ratio (CAR) in severe acute pancreatitis (SAP) and the value of SAP and concomitant acute liver injury (ALI).Methods:Total of 195 patients with AP from June 2021 to December 2022 from 374 patients were screened for inclusion in the study and were divided into non-severe acute pancreatitis (NSAP) and SAP groups. The ALI group was divided into non-acute liver injury (NALI) and ALI groups according to ALI criteria, and then into hepatocellular ALI subgroup, cholangiocellular ALI subgroup and mixed ALI subgroup. Laboratory tests for procalcitonin (PCT), C-reactive protein (CRP), albumin and platelet (PLT) were completed within 48 h. Risk factors for SAP, ALI and each subgroup of ALI were analysed by binary logistic regression. Subject work characteristic (ROC) curves were plotted and the optimal thresholds for PPR and CAR were calculated. The predictive value of PPR, CAR and their combination for SAP, ALI and each type of ALI was determined.Results:The AUCs for predicting SAP by plotting ROC curves and calculating the bedside index score of acute pancreatitis severity (BISAP score), PPR, CAR, PPR combined with CAR, PPR combined with BISAP score, CAR combined with BISAP score and combined PPR, CAR and BISAP score were 0.82, 0.85, 0.79 and 0.86. The areas under the ROC curves for PPR, CAR and combined prediction of ALI were 0.81, 0.85 and 0.88, respectively; the areas under the ROC curves for PPR, CAR and combined prediction of hepatocellular ALI were 0.93, 0.77 and 0.92, respectively; and the areas under the ROC curves for PPR, CAR and combined prediction of cholangiocellular ALI were 0.76, 0.76 and 0.77, respectively. The area under the ROC curves for PPR, CAR and combined prediction of mixed ALI were 0.83, 0.76 and 0.82Conclusions:Elevated PPR and CAR are risk factors for SAP and for the development of ALI in AP. PPR has better predictive value than CAR for hepatocellular and mixed ALI, and CAR has better predictive value than PPR for cholangiocellular ALI.
6.Predictive value of FAR, CAR and PLR in hyperlipidemic acute pancreatitis
Qiaofang WANG ; Chaopeng MEI ; Yaodong SONG ; Yanna LIU ; Dejian LI ; Mengwei CUI ; Qianqian HE ; Huihui LI ; Haifeng WANG ; Changju ZHU
Chinese Journal of Emergency Medicine 2024;33(10):1376-1382
Objective:To investigate the value of fibrinogen to albumin ratio (FAR), creatinine to albumin ratio (CAR) and platelet to lymphocyte ratio (PLR) in predicting the poor prognosis of hyperlipidemic acute pancreatitis (HLAP).Methods:Clinical data of HLAP patients admitted to the hospital from January 2021 to January and December 2023 were retrospectively collected. According to the prognosis, the patients were divided into two groups: good prognosis group and poor prognosis group.The independent risk factors of HLAP in different prognostic groups were obtained by multivariate Logistic regression analysis. Receiver operating characteristic (ROC) curves were plotted to evaluate the prognostic value of FAR, CAR and PLR alone and in combination.Results:A total of 118 patients with HLAP were included, including 69 patients with good prognosis and 49 patients with poor prognosis.The difference of heart rate, lymphocyte, triglyceride, albumin, creatinine, urea nitrogen, blood calcium, blood glucose, C-reactive protein, procalcitonin, fibrinogen, FAR, CAR, PLR, Bedside indicator of acute pancreatitis Severity score, Acute Physiology and Chronic Health status score, hospitalization time assessment between the two groups was statistically significant ( P<0.05). Multivariate Logistic regression analysis showed that FAR (odds ratio ( OR) = 25.949, 95% confidence interval (95% CI):3.190 ~ 211.080, P = 0.002), CAR ( OR = 1.453, 95% CI:1.095 ~ 1.928, P = 0.010) and PLR ( OR = 1.005, 95% CI: 1.001 ~ 1.009, P = 0.020) were independent risk factors for poor prognosis in HLAP patients. ROC curve analysis showed that the area under the ROC curve (AUC) of FAR, CAR and PLR to predict poor prognosis of HLAP patients were 0.823, 0.781 and 0.652, respectively.The AUC of FAR combined with CAR, FAR combined with PLR and CAR combined with PLR were 0.840, 0.845 and 0.849, respectively.The combined ability of FAR, CAR and PLR to predict poor prognosis in HLAP patients was (AUC=0.875,95% CI:0.814 ~ 0.937). When the cut-off value was 0.387, the sensitivity was 83.7%, and the specificity was 79.7%. Conclusions:The prognostic value of FAR, CAR and PLR in HLAP patients is better than that of single or pairwise combination.
7.Study of a patient with Myelodysplastic/myeloproliferative neoplasm with co-morbid neutrophilia and a novel NCOR1: : GLYR1 fusion gene
Yutian LEI ; Xiaoli ZHAO ; Huihui ZHAO ; Yu CHENG ; Shuai WANG ; Jianyong LI ; Yu ZHU
Chinese Journal of Medical Genetics 2024;41(4):404-410
Objective:To explore the genetic background for a patient with refractory myelodysplastic/myeloproliferative neoplasm (MDS/MPN) with co-morbid neutrophilia patient.Methods:A MDS/MPN patient who was admitted to the First Affiliated Hospital of Nanjing Medical University in May 2021 was selected as the study subject. RNA sequencing was carried out to identify fusion genes in his peripheral blood mononuclear cells. Fusion gene sequence was searched through transcriptome-wide analysis with a STAR-fusion procedure. The novel fusion genes were verified by quantitative real-time PCR and Sanger sequencing.Results:The patient, a 67-year-old male, had progressive thrombocytopenia. Based on the morphological and molecular examinations, he was diagnosed as MDS/MPN with co-morbid neutropenia, and was treated with demethylating agents and Bcl-2 inhibitors. Seventeen months after the diagnosis, he had progressed to AML. A novel fusion gene NCOR1: : GLYR1 was identified by RNA-sequencing in his peripheral blood sample, which was verified by quantitative real-time PCR and Sanger sequencing. The patient had attained morphological remission after a DCAG regimen (a combinatory chemotherapy of decitabine, cytarabine, aclarubicin and granulocyte colony-stimulating factors) plus Chidamide treatment. A significant decrease in the NCOR1: : GLYR1 expression was revealed by quantitative real-time PCR at post-chemotherapy evaluation. Conclusion:NCOR1: : GLYR1 gene is considered as the pathogenic factor for the MDS/MPN patient with neutropenia.
8.Scoping review of continuous bladder irrigation practice following transurethral resection of the prostate
Jingxiong WU ; Wenwen CHEN ; Huihui FANG ; Chen ZHU ; Xiangyun LU
Chinese Journal of Modern Nursing 2024;30(21):2929-2935
Objective:To comprehensively and systematically collect and review domestic and international studies on the practices of continuous bladder irrigation (CBI) following transurethral resection of the prostate (TURP), including the topics, research methods, and evaluation techniques used in these studies, and to provide clinical nursing staff with a reference.Methods:Nine Chinese and English databases were systematically searched, including CNKI, Wanfang, SinoMed, VIP, Embase, Cochrane Library, PubMed, CINAHL, and Web of Science. The search covered all literature up to April 10, 2023. The practices of CBI were extracted and systematically analyzed, and the results were reported in a standardized manner using a scoping review methodology.Results:A total of 23 eligible studies were included, comprising two English articles and 21 Chinese articles. The types of studies included 12 randomized controlled trials and 11 quasi-experimental studies. The interventions were categorized into single and comprehensive measures, including methods such as adjusting the temperature and flow rate of bladder irrigation, selection of indwelling catheters, traditional Chinese medicine-based nursing, perioperative psychological care, and rapid recovery protocols. The evaluation metrics used in the studies were classified into three levels: symptom indicators, process indicators, and management indicators.Conclusions:Research on CBI following TURP has achieved a certain scale, with a variety of intervention methods currently available. However, there is still a lack of high-quality evidence. Future studies should focus on enhancing research design, developing standardized quality assessment tools for bladder irrigation, and improving innovative irrigation devices to enhance clinical nursing quality.
9.Chest CT quantitative parameters in patients with acute exacerbation of chronic obstructive pulmonary disease:Correlations with blood eosinophil level
Lu YANG ; Yadan SHENG ; Kai YANG ; Liyu HE ; Huihui GU ; Guoqing SUN ; Weiting CHEN ; Bingjie ZHU ; Yanrong CHEN ; Chenwang JIN
Chinese Journal of Medical Imaging Technology 2024;40(8):1189-1193
Objective To observe the correlations of chest CT quantitative parameters in patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)with blood eosinophil(EOS)level.Methods Chest CT data of 162 AECOPD patients with elevated eosinophils were retrospectively analyzed.The patients were divided into low EOS group(n=105)and high EOS group(n=57)according to the absolute counting of blood EOS.The quantitative CT parameters,including the number of whole lung bronchi and the volume of blood vessels,low-attenuation area percentage(LAA%)of whole lung,of left/right lung and each lobe of lung,as well as the luminal diameter(LD),wall thickness(WT),wall area(WA)and WA percentage of total bronchial cross-section(WA%)of grade 3 to 8 bronchi were compared between groups.Spearman correlations were performed to analyze the correlations of quantitative CT parameters with blood EOS level.Results LAA%of the whole lung,of the left/right lung and each lobe of lung,as well as of the upper lobe of right lung LDgrade4,middle lobe of right lung WTgrade5,upper lobe of right lung WAgrade4,middle lobe of right lung WAgrade5 and lower lobe of left lung WAgrade3 in low EOS group were all higher than those in high EOS group(all P<0.05).Except for the upper lobe of right lung LDgrade4,the above quantitative CT indexes being significant different between groups were all weakly and negatively correlated with blood EOS level(r=-0.335 to-0.164,all P<0.05).Conclusion Chest CT quantitative parameters of AECOPD patients were correlated with blood EOS level,among which LAA%,a part of WT and WA were all weakly negatively correlated with blood EOS level.
10.Construction of a chimeric antigen receptor CAR THP -1 cell line targeting HER2
Yizhao Chen ; Lihua Liu ; Xiangling Zhu ; Huihui Wang ; Xuming Wu ; Xuewen Tan ; Yilong Fang ; Haifeng Jiang ; Zhen Xu ; Wei Wei ; Jiajie Tu
Acta Universitatis Medicinalis Anhui 2023;58(3):352-357
Objective:
To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected.
Methods :
CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) .CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro.The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression,and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ,and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry.
Results :
CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells,flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P<0. 01) .
Conclusion
In this experiment,CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ,and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.


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