Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8％(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67％similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.

Objective The mouse autologous tumor model H22 is more valuable for tumor immunological-related research.This paper aims to establish mouse hepatic tumor cell line(H22-luc2-tdT)that stably express the tan-dem-dimer tomato(tdTomato)and luciferase genes.Establish an in vivo imaging model of cell line derived trans-planted tumors.Methods Using transplanted H22 tumor tissue,primary culture and continuous passage in vitro were performed to establish a continuous cell line.Cell proliferation,chromosome analysis,organoid culture,tumorigenicity,HE and ICH of aFP,CK7,CK15 were performed to charaterize the cell line.Then the luc2-tdT plasmid was transfected into H22 cells of P22,flow cytometry and in vitro/in vivo imaging were employed to screen and verify fluorescence expression.Mycoplasma detection and species verification of the established cell lines were performed.Results The H22 cells had been continuously passaged over 50 times.The cells of passsge 22(P22)were transplanted subcutaneously and intraperitoneally into C57 and Kunming mice,with a 100%tumor formation.The HE morphology of subcutaneous transplanted tumor were consistent with the original tumor.CK+/AFP+proved that it was of liver cancer origin.The H22 cells were hypo-triploid with a modal number of 40-44 chromosomes and telocentromeres,verifing its mouse origin.The latent phase for in vitro growth of H22 lasted from d0 to d3,while the exponential phaes d3 to d5,and reach plateou at d6.Successful transfection of H22 cells with the luc2-tdT were observed with in vitro/in vivo 100%fluorescence positivity,thus named H22-luc2-tdT.The transplanted tumor tissue of H22 cells could be primarily cultured to form organoids.The detection of Mycoplasma was negative,and its mouse origin confirmed by PCR.Conclusions H22 and H22-luc2-tdT cell lines are established and characterized,which can be used for the establishment and application of in vitro and in vivo liver cancer research and metastatic cell tracking.These cell lines are deposited at and can obtain from the National Biomedical Cell Resource Center(http://www.cellresource.cn).

Objective To establish primary and simian virus 40(SV40)T antigen transformed human vascular en-dothelial cell models,and provide available resources for endothelial research.Methods Human umbilical vein endothelial cells(HUVEC),human umbilical artery endothelial cells(HUAEC),great saphenous vein endothelial cells(GSVEC)and endothelial cells form endometrium and liver tissue were isolated and cultured respectively.Then,the primary endothelial cells were transformed by lentivirus containing SV40 big T and small T antigens,and continuously subcultured in vitro.The expression of CD31 was detected by flow cytometry,species identification-and mycoplasma detection by PCR,and cell identity was identified by STR detection.The transformed ECs were checked for HLA types.Some of them were tested for RNA expression profile and infected by Cas9 lentivirus to es-tablish stable clones.Results Totally 187 cell lines of transformed HUVEC,1 of transformed HUAEC,5 of trans-formed GSVEC,1 of transformed endothelial cells from endometrium and 1 of transformed endothelial cells from liv-er tissue,and 9 monoclonal HUVEC cell lines stably expressing Cas9 protein were established.All the transformed umbilical endothelial cells were CD31 positive ranging from 20%-90%for 20 cases,while for the rest 168 cases the positive rate was more than 90%.RNA expression revealed stable activation of cell proliferation(cell cycle and DNA synthesis).Their species were identified as human origin.The STR results were consistent with those of the primary culture and unique,and there was no mycoplasma contamination.All these cells could be obtained with the sharing services of National Science and Technology Infrastructure,the National Biomedical Cell-line Resource cen-ters(NSTI-BMCR).Conclusions A series of primary and SV40 T antigen transformed human vascular endothelial cell models have been established,which provide a tool for the study of cardiovascular diseases,inflammation,tumors and immune-related diseases.

To review the clinical characteristics, short-term outcomes, and risk factors of patients with infective endocarditis(IE) who underwent surgical treatment at a single center, and to summarize treatment experience.

Objective:To investigate the current status of subjective well-being among stroke patients, and to explore the pathways and effects of influencing factors using structural equation model, so as to provide reference for improving subjective well-being among stroke patients.Methods:From July to November 2024, the stroke patients admitted to the First Affiliated Hospital of Bengbu Medical University, the Second Affiliated Hospital of Bengbu Medical University, Hefei First People′s Hospital were selected by convenience sampling method. A cross-sectional survey was conducted using a general demographic questionnaire, General Well-Being Scale, Cognitive Reserve Index questionnaire, Social Support Rating Scale, Stroke Symptom Cluster Scale, and FRAIL Scale, and AMOS 26.0 was used to analyse the pathways and effects of influencing factors of subjective well-being.Results:A total of 435 questionnaires were collected, 410 were valid.Among 410 cases, 266 case were males, 144 were females, with an age of (65.96 ± 12.15) years. The subjective well-being scores of stroke patients were (72.58 ± 11.66) points. Cognitive reserve and social support were positively correlated with subjective well-being ( r = 0.517, 0.554, both P<0.01), while symptom burden and frailty were negatively correlated with subjective well-being ( r = -0.687, -0.670, both P<0.01). Path analysis showed that symptom burden, frailty, cognitive reserve, and social support had a direct impact on subjective well-being (path coefficients were -0.500, -0.266, 0.148, and 0.144, respectively, all P<0.05), while cognitive reserve, social support, and symptom burden had an indirect impact on subjective well-being (path coefficients were 0.287, 0.249, and 0.108, respectively, all P<0.05). Conclusions:The subjective well-being of stroke patients is influenced by multiple factors, with symptom burden being an important factor affecting subjective well-being. Intervention strategies such as improving cognitive reserve, strengthening social support systems, and preventing frailty can improve the subjective well-being of patients.

Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8％(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67％similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.

Objective:To investigate the current status of subjective well-being among stroke patients, and to explore the pathways and effects of influencing factors using structural equation model, so as to provide reference for improving subjective well-being among stroke patients.Methods:From July to November 2024, the stroke patients admitted to the First Affiliated Hospital of Bengbu Medical University, the Second Affiliated Hospital of Bengbu Medical University, Hefei First People′s Hospital were selected by convenience sampling method. A cross-sectional survey was conducted using a general demographic questionnaire, General Well-Being Scale, Cognitive Reserve Index questionnaire, Social Support Rating Scale, Stroke Symptom Cluster Scale, and FRAIL Scale, and AMOS 26.0 was used to analyse the pathways and effects of influencing factors of subjective well-being.Results:A total of 435 questionnaires were collected, 410 were valid.Among 410 cases, 266 case were males, 144 were females, with an age of (65.96 ± 12.15) years. The subjective well-being scores of stroke patients were (72.58 ± 11.66) points. Cognitive reserve and social support were positively correlated with subjective well-being ( r = 0.517, 0.554, both P<0.01), while symptom burden and frailty were negatively correlated with subjective well-being ( r = -0.687, -0.670, both P<0.01). Path analysis showed that symptom burden, frailty, cognitive reserve, and social support had a direct impact on subjective well-being (path coefficients were -0.500, -0.266, 0.148, and 0.144, respectively, all P<0.05), while cognitive reserve, social support, and symptom burden had an indirect impact on subjective well-being (path coefficients were 0.287, 0.249, and 0.108, respectively, all P<0.05). Conclusions:The subjective well-being of stroke patients is influenced by multiple factors, with symptom burden being an important factor affecting subjective well-being. Intervention strategies such as improving cognitive reserve, strengthening social support systems, and preventing frailty can improve the subjective well-being of patients.

Objective To systematically review the incidence and influencing factors of refeeding syndrome(RFS)in critically ill patients,and provide references for early identification of RFS and formulation of preventive measures.Methods Computerized searches were conducted for studies on RFS in critically ill patients in the databases of China National Knowledge Infrastructure(CNKI),Wanfang,VIP,CBM,PubMed,Embase,Web of Science,CINAHL,Cochrane Library from inception to May 29th,2024.Data analysis was performed using Stata 16.0 software.Results A total of 29 articles with 5 720 participants were included.The Meta-analysis showed that the incidence of RFS in critically ill patients was 33.68％.The subgroup analysis showed that the incidence of RFS in critically ill patients was higher in studies conducted in 2020 or later(38.22％),in the Americas(36.39％),and with only electrolyte changes as the diagnostic basis(37.51％).Risk factors for RFS in critically ill patients included higher acute physiological and chronic health evaluation Ⅱ scores(OR=1.41),higher sequential organ failure assessment scores(OR=1.29),initiation of feeding within 48 h of ICU admission(OR=3.36),age ≥60 years(OR＝2.82),diabetes mellitus(OR=3.53),pre-albumin concentration＜150 g/L(OR=5.53),albumin concentration＜30 g/L(OR=3.26),caloric intake＞25％standard calories(OR=2.86),enteral solution temperature of 36～38 ℃(OR=2.32),feeding rate＞50 ml/h(OR=3.76),fasting time ≥2 d before feeding(OR=2.46),history of alcoholism(OR=2.64).Conclusion The incidence of RFS in critically ill patients is high and there are many influencing factors.Nurses should improve their awareness and attention to RFS,accurately identify high-risk groups and risk factors,and adopt a multidisciplinary collaborative model to develop whole-course,detailed and personalized intervention measures to prevent RFS.

Objective To establish human and mouse pancreatic cancer cell strains stably expressing Cas9 protein,green fluorescent protein,red fluorescent proteins,luciferase-tdTomato,and to validate the activity of luciferase and gene editing of Cas9 function for pancreatic cancer research using luciferase and CRISPR/Cas9 system.Methods In human pancreatic cancer cells(AsPC-1,CFPAC-1,HPAC,BxPC-3,HS 766T,MIA PaCa-2,PANC-1,and SW 1990),and mouse pancreatic cancer cell(Pan02),the cells were infected with Cas9-expressing plas-mid pLv-EF1α-Cas9m1.1-Puro,and single-cell clones were selected for culture and expansion.After extracting the total protein,Western blot verified the expression level of Cas9;Infected with fluorescent protein expression plasmids pLv-EF1α-EGFP,pLv-EF1α-mCherry,pLv-EF1α-tdTomato,pLv-EF1α-Luc2-tdT,and selected single cell clones stably expressing fluorescent proteins were cultured and amplified under fluorescence microscope.Cas9 stable expression cell line was selected to be infected with pLv-EF1α-Luc2-tdT,and the mono-clonal culture of stable expression of fluorescent proteins was selected for expansion under fluorescence micro-scope.One of the cell lines were selected to be infected with Lv-EF1a-mCherry,and the mCherry-positive cells were sorted out by flow cytometry,and then the guide RNA of mCherry gene was then infected by lentivirus to tar-get the mCherry gene,and after cell expansion,mCherry knockdown was detected by fluorescence microscope observation and flow cytometry;5 BALB/c Nude mice were subcutaneously inoculated with MIA PaCa-2-Luc2-tdT cells(1.0×107/cells each),and imaged in vivo after 36 days.Results 48 human pancreatic cancer cell strains with stable Cas9 expression were screened(including 23 cells expressing Cas9m1.1,25 cells expressing Cas9m1.1-Luc2-tdT),33 pancreatic cancer cell strains with stable expression of fluorescent proteins were screened(8 cells expressing EGFP,7 expressing mCherry,and 9 each expressing Luc2-tdT and tdTomato).Cells expressing mCherry and Cas9 were infected with mCherry gRNA and mCherry was knocked down.In vivo imaging showed that both bioluminescence and fluorescence luminescence were present in MIA PaCa-2 cells ex-pressing Luc2-tdT.Conclusions 33 pancreatic cancer cell strains with stable expression of fluorescent proteins are successfully established,in which the Luc2-tdT-expressing cell strains have luciferase activity;48 pancreatic cancer cell strains with stable expression of Cas9 are successfully established,and the Cas9 protein has gene edi-ting activity,gene editing activity varies depending on the original cell strains.

Objective To explore the accuracy and influencing factors of contrast-enhanced spectral mammography(CESM)and MRI in the dimension measurement of breast lesions.Methods A retrospective analysis was performed on 104 patients with breast lesions who underwent CESM and MRI examinations within one week.The pathological size was used as the gold standard to com-pare the accuracy of the two measurement methods.Logistic regression analysis was used to compare the influencing factors of the measurement accuracy of CESM and MRI.Results The measured values of CESM,MRI and pathology were 1.70 cm(1.20-2.70 cm),1.65 cm(1.20-2.28 cm),1.75 cm(1.00-2.50 cm),respectively,and there were no statistically significant differences between the two measurement methods and the pathological results(P＞0.05).Bland-Altman plot analysis showed that 95.2％(99/104)of the differences in CESM fell within the consistency interval,and 93.3％(97/104)of the differences in MRI fell within the consistency interval.Logistic regression analysis showed that the independent risk factors affecting the accuracy of CESM and MRI measurement were lesion size and enhancement type.Conclusion Both CESM and MRI show good accuracy.As a potential alternative to MRI technology,CESM can be used for imaging evaluation of breast lesion size.