To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objective:To construct an evaluation indicator system for power system of otolaryngology surgery,so as to provide references for the configuration of surgical power devices of medical institutions.Methods:Literature review and brainstorming were used to analyze existing literature related to surgical power.Combined with expert opinions and clinical demands,an evaluation indicator system was initially proposed.The Delphi method was adopted to determine the evaluation indicators of power system of otolaryngology surgery.The analytic hierarchy process(AHP)method was used to determine the evaluation indicator system of the power system of otolaryngology surgery,which were constructed by weight of each indicator.Results:The evaluation indicator system of power system of otolaryngology surgery included 5 first-level indicators(integrity of medical equipment,products'performance indicators,safety,clinical application effect,and after-sales service guarantee)and 49 second-level indicators under the first-level indicators.In the first-level indicators,equipment's safety had the highest weight(20.48%).In the second-level indicators,the top three of the combined weights were respectively integrity of equipment's main device(7.19%),accessory's integrity(7.03%),and identification's integrity(6.03%).Conclusion:The evaluation index system for otolaryngological surgical power systems clarifies the core dimensions,specific indicators and relative importance of the evaluation,and can be applied to the procurement and selection of surgical power devices in medical institutions,performance testing,clinical effect evaluation and other aspects.

Objective:To explore the accuracy of parameters of anterior segment measurement assessment system and optical biometer in predicting lens in implantable collamer lens(ICL)implantation.Methods:This study retrospectively analyzed the examination data of patients who underwent ICL surgery at the department of ophthalmology,Army Characteristic Medical Center from January 2020 to July 2024.The measured data of the chamber corner parameters,which included anterior chamber depth(ACD),central corneal thickness(CCT),white to white(WTW),flat keratometry(K1)and steep keratometry(K2),of 74 patients(134 eyes)who underwent simultaneous measurements of the IOLMaster 700 optical biometer and the Pentacam HR anterior segment measurement assessment system were selected.The expected spherical equivalent(EXP SPH),expected cylinder(EXP CYL),and expected spherical equivalent(EXP SEQ)were calculated,and the differences,correlations and consistencies of these data were analyzed.Results:There were significant differences in ACD,CCT,WTW,K1 and K2 between the IOLMaster 700 and Pentacam HR in measuring myopic patients(t=12.928,6.202,-37.877,-13.743,-12.007,P<0.001).All measured data showed high correlation(r=0.983,0.984,0.961,0.996,0.994,P<0.001).Bland-Altman consistency test showed that the measured values by 3%-6%were outside the limits of 95%consistency,and the differences among the EXP SPH of two instruments and the postoperative actually measured results of spherical lens(SPH),among the EXP CYL of two instruments and the postoperative actually measured results of cylinder lens(CYL),and among the EXP SEQ of two instruments and the postoperative actual measured results of spherical equivalent(SEQ)were statistically significant(x2=20.927,7.451,25.459,P<0.05).The average differences between Pentacam HR and IOLMaster 700 were respectively(0.024±0.491)D and(0.009±.491)D in predicting SPH,and were respectively(-0.05±0.476)D and(-0.049±0.475)D in predicting CYL,and were respectively(-0.001±0.458)D and(-0.016±0.461)D in predicting SEQ.Conclusion:During selection process of ICL surgery for lens,clinicians can reasonably choose Pentacam HR and IOLMaster 700 to calculate lens according to the actual requirements of the patients.

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Phage immunoprecipitation sequencing (PhIP-Seq) is a high-throughput and low-cost method for analyzing the specific binding of target proteins to peptide libraries. The method uses oligonucleotide library synthesis (OLS) to encode proteome-scale peptide libraries for display on phages, and then immunoprecipitates these library phages with target proteins (such as antibodies) for subsequent analysis by high-throughput DNA sequencing. PhIP-Seq enables the screening of peptide targets that react specifically with hundreds of proteins or pathogens. PhIP-Seq has been successfully applied in various fields such as disease detection, screening of autoimmune disease biomarkers, vaccine development, and allergen detection, becoming a high-throughput diagnostic technology. This article systematically describes the development, applications, and result evaluation of PhIP-Seq, in order to gain a more comprehensive understanding of the application and future development prospects of this technology in various fields.
Peptide Library ; Humans ; Immunoprecipitation/methods* ; High-Throughput Nucleotide Sequencing/methods* ; Bacteriophages/genetics*

Objective:To explore the cardioprotective effects of icariin (ICA) against radiation-induced cardiac disease (RICD) in C57BL/6 mice.Methods:A total of 48 female C57BL/6J mice aged 6-8 weeks were randomly divided into three groups: the control group (CON), the irradiation group (IR), and the irradiation combined with icariin group (IR+ ICA), with 16 mice in each group. The IR and IR+ ICA groups received a single cardiac irradiation at a dose of 30 Gy, while the CON group received no radiation treatment. The IR+ ICA group was treated with ICA (70 mg·kg -1·d -1) two weeks before irradiation until the end of the experiment through intragastric administration. In contrast, the CON and IR groups were treated with an equal volume of vehicle solution (0.5% sodium carboxymethyl cellulose, NaCMC) via intragastric administration. The mice′s mental status, food intake, body weight, and survival rates were monitored during the experiment. At two weeks post-irradiation, the venous blood of the mice was collected and serum was separated for the enzyme-linked immunosorbent assays (ELISA) of creatine kinase MB isoenzyme (CK-MB) and cardiac troponin T (cTnT/TNNT2). At 12 weeks post-irradiation, the cardiac function of the mice was assessed using echocardiography. After the mice were euthanized under anesthesia, the histopathological changes and fibrosis degree of their myocardial tissues were assessed using hematoxylin and eosin (HE) and Masson′s trichrome staining, followed by the calculation of collagen volume fraction (CVF). The differential gene expression of brain natriuretic peptide (BNP), transforming growth factor-β (TGF-β), and interleukin-6 (IL-6) in the cardiac tissues of the mice was detected using real-time reverse transcription-polymerase chain reaction (RT-PCR). Apoptosis-related proteins and proteins associated with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway were determined using Western blotting. The survival curves of the mice were plotted using Kaplan-Meier, and the survival differences of the mice among various groups were compared using the log-rank test. Results:After irradiation, the mice in the IR group showed lethargy, as well as decreased food intake and activity, while these symptoms in the IR+ ICA group were significantly alleviated. At two weeks post-irradiation, the CK-MB and cTnT levels of the IR group were significantly elevated compared with the CON group ( t = 5.28, 8.89, P < 0.01). At 12 weeks post-irradiation, the mice in the IR group exhibited significantly decreased body weight ( t = 2.47, P < 0.05) and decreased survival rates ( HR = 8.25, 95% CI: 1.157-58.770, P < 0.05) compared with the CON group. Echocardiography revealed that the IR group featured decreased left ventricular ejection fraction (EF), decreased fractional shortening (FS), and increased left ventricular end-diastolic diameter (LVDD) compared with the CON group ( t = 7.02, 4.45, P < 0.05). Histopathological examination revealed that the IR group suffered from cardiomyocyte edema, disordered arrangement, and increased fibrosis, with an elevated CVF. The IR group exhibited significantly upregulated gene expression of BNP, TGF-β, and IL-6 in cardiac tissues compared with the CON group ( t = 4.23, 6.39, 4.61, P < 0.05). After-irradiation, the IR group exhibited upregulated apoptosis-related proteins Cleaved Caspase-3 and Bax ( t = 6.29, 9.54, P < 0.05), decreased Bcl-2 expression ( t = 8.20, P < 0.001), and decreased phosphorylation levels of PI3K and Akt ( t = 6.47, 3.42, P < 0.001). The symptoms of the mice were partially ameliorated after treatment with ICA. Specifically, the mice in the IR+ ICA group exhibited higher body weight ( t = 5.13, P < 0.001) and significantly higher survival rates ( HR = 0.121, 95% CI: 0.017-0.864, P < 0.05) than the IR group. Compared to the IR group, the IR+ ICA group showed elevated cardiac function indicators EF and FS( t = 3.23, 3.05, P < 0.05), and reduced LVDD ( t = 3.02, P < 0.05). The histopathological analysis revealed mitigated edema and disordered arrangement of cardiomyocytes in the IR+ ICA group. Furthermore, the IR+ ICA group exhibited significantly lower BNP, TGF-β, and IL-6 expression levels than the IR group ( t = 2.83, 4.15, 2.96, P < 0.05). The expression of apoptosis-related proteins Cleaved Caspase-3 and Bax was lower ( t = 3.23, 3.24, P < 0.05), Bcl-2 expression was higher ( t = 5.92, P < 0.001), and restored phosphorylation levels of PI3K and Akt ( t = 2.89, 8.35, P < 0.001). Conclusions:Icariin has protective effects against the RICD. It alleviates cardiomyocyte apoptosis possibly by upregulating the phosphorylation levels of PI3K and Akt.

Objective:To explore the cardioprotective effects of icariin (ICA) against radiation-induced cardiac disease (RICD) in C57BL/6 mice.Methods:A total of 48 female C57BL/6J mice aged 6-8 weeks were randomly divided into three groups: the control group (CON), the irradiation group (IR), and the irradiation combined with icariin group (IR+ ICA), with 16 mice in each group. The IR and IR+ ICA groups received a single cardiac irradiation at a dose of 30 Gy, while the CON group received no radiation treatment. The IR+ ICA group was treated with ICA (70 mg·kg -1·d -1) two weeks before irradiation until the end of the experiment through intragastric administration. In contrast, the CON and IR groups were treated with an equal volume of vehicle solution (0.5% sodium carboxymethyl cellulose, NaCMC) via intragastric administration. The mice′s mental status, food intake, body weight, and survival rates were monitored during the experiment. At two weeks post-irradiation, the venous blood of the mice was collected and serum was separated for the enzyme-linked immunosorbent assays (ELISA) of creatine kinase MB isoenzyme (CK-MB) and cardiac troponin T (cTnT/TNNT2). At 12 weeks post-irradiation, the cardiac function of the mice was assessed using echocardiography. After the mice were euthanized under anesthesia, the histopathological changes and fibrosis degree of their myocardial tissues were assessed using hematoxylin and eosin (HE) and Masson′s trichrome staining, followed by the calculation of collagen volume fraction (CVF). The differential gene expression of brain natriuretic peptide (BNP), transforming growth factor-β (TGF-β), and interleukin-6 (IL-6) in the cardiac tissues of the mice was detected using real-time reverse transcription-polymerase chain reaction (RT-PCR). Apoptosis-related proteins and proteins associated with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway were determined using Western blotting. The survival curves of the mice were plotted using Kaplan-Meier, and the survival differences of the mice among various groups were compared using the log-rank test. Results:After irradiation, the mice in the IR group showed lethargy, as well as decreased food intake and activity, while these symptoms in the IR+ ICA group were significantly alleviated. At two weeks post-irradiation, the CK-MB and cTnT levels of the IR group were significantly elevated compared with the CON group ( t = 5.28, 8.89, P < 0.01). At 12 weeks post-irradiation, the mice in the IR group exhibited significantly decreased body weight ( t = 2.47, P < 0.05) and decreased survival rates ( HR = 8.25, 95% CI: 1.157-58.770, P < 0.05) compared with the CON group. Echocardiography revealed that the IR group featured decreased left ventricular ejection fraction (EF), decreased fractional shortening (FS), and increased left ventricular end-diastolic diameter (LVDD) compared with the CON group ( t = 7.02, 4.45, P < 0.05). Histopathological examination revealed that the IR group suffered from cardiomyocyte edema, disordered arrangement, and increased fibrosis, with an elevated CVF. The IR group exhibited significantly upregulated gene expression of BNP, TGF-β, and IL-6 in cardiac tissues compared with the CON group ( t = 4.23, 6.39, 4.61, P < 0.05). After-irradiation, the IR group exhibited upregulated apoptosis-related proteins Cleaved Caspase-3 and Bax ( t = 6.29, 9.54, P < 0.05), decreased Bcl-2 expression ( t = 8.20, P < 0.001), and decreased phosphorylation levels of PI3K and Akt ( t = 6.47, 3.42, P < 0.001). The symptoms of the mice were partially ameliorated after treatment with ICA. Specifically, the mice in the IR+ ICA group exhibited higher body weight ( t = 5.13, P < 0.001) and significantly higher survival rates ( HR = 0.121, 95% CI: 0.017-0.864, P < 0.05) than the IR group. Compared to the IR group, the IR+ ICA group showed elevated cardiac function indicators EF and FS( t = 3.23, 3.05, P < 0.05), and reduced LVDD ( t = 3.02, P < 0.05). The histopathological analysis revealed mitigated edema and disordered arrangement of cardiomyocytes in the IR+ ICA group. Furthermore, the IR+ ICA group exhibited significantly lower BNP, TGF-β, and IL-6 expression levels than the IR group ( t = 2.83, 4.15, 2.96, P < 0.05). The expression of apoptosis-related proteins Cleaved Caspase-3 and Bax was lower ( t = 3.23, 3.24, P < 0.05), Bcl-2 expression was higher ( t = 5.92, P < 0.001), and restored phosphorylation levels of PI3K and Akt ( t = 2.89, 8.35, P < 0.001). Conclusions:Icariin has protective effects against the RICD. It alleviates cardiomyocyte apoptosis possibly by upregulating the phosphorylation levels of PI3K and Akt.

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objective:To investigate the imaging features of single hip joint in patients with acetabular labrum injury, and to analyze the clinical value of 3.0T MRI scanning in the diagnosis of acetabular labrum injury.Methods:Clinical data of 76 patients with high suspicion of acetabular labrum injury and hip arthroscopy during clinical physical examination in Peking University Third Hospital Qinhuangdao Hospital from January 2020 to December 2023 were retrospectively collected. All patients underwent 3.0T MRI scan of single hip joint before surgery. The type, location and degree of acetabular labrum injury were evaluated by imaging diagnosis, and the diagnostic efficacy of 3.0T MRI single hip scan for acetabular labrum injury was analyzed with the hip arthroscopy as the " gold standard" .Results:Hip arthroscopy showed that 63 patients with acetabular labrum injury had 87 lesions in 228 labrum regions, including 56 lesions in the anterior labrum, 25 lesions in the outer upper labrum, and 6 lesions in the posterior labrum. The injury types were 141 without tear, 29 with parenchymal tear of acetabular labrum and 58 with cartilage junction of acetabular labrum. MRI scan showed changes in high signal within the labrum of the acetabulum, and there were high signal shadows between the labrum base and the acetabular cartilage, and the signal was uneven. The sensitivity, specificity and accuracy of single hip MRI scanning in the diagnosis of acetabular labrum injury were 85.71%, 84.62% and 85.53%, respectively. The lesions of anterior pelvis and lip were 80.36%, 75.00% and 78.95%, respectively. The injuries of external upper pelvis and lip were 72.00%, 80.39% and 77.63%, respectively. The injuries of posterior pelvis and lip were 100.00%, 95.71% and 96.05%, respectively. The sensitivity, specificity and accuracy of single hip MRI in the diagnosis of parenchymatous tear of acetabulum labrum were 100.00%, 94.83% and 96.55%, respectively, which was in good agreement with that of hip arthroscopy (Kappa=0.924).Conclusions:3.0T MRI scan of single hip can effectively diagnose the injury of acetabular pelvis and lip, and has high diagnostic value in the differential diagnosis of different parts and types of acetabular pelvis and lip injury.

Objective:To investigate the imaging features of single hip joint in patients with acetabular labrum injury, and to analyze the clinical value of 3.0T MRI scanning in the diagnosis of acetabular labrum injury.Methods:Clinical data of 76 patients with high suspicion of acetabular labrum injury and hip arthroscopy during clinical physical examination in Peking University Third Hospital Qinhuangdao Hospital from January 2020 to December 2023 were retrospectively collected. All patients underwent 3.0T MRI scan of single hip joint before surgery. The type, location and degree of acetabular labrum injury were evaluated by imaging diagnosis, and the diagnostic efficacy of 3.0T MRI single hip scan for acetabular labrum injury was analyzed with the hip arthroscopy as the " gold standard" .Results:Hip arthroscopy showed that 63 patients with acetabular labrum injury had 87 lesions in 228 labrum regions, including 56 lesions in the anterior labrum, 25 lesions in the outer upper labrum, and 6 lesions in the posterior labrum. The injury types were 141 without tear, 29 with parenchymal tear of acetabular labrum and 58 with cartilage junction of acetabular labrum. MRI scan showed changes in high signal within the labrum of the acetabulum, and there were high signal shadows between the labrum base and the acetabular cartilage, and the signal was uneven. The sensitivity, specificity and accuracy of single hip MRI scanning in the diagnosis of acetabular labrum injury were 85.71%, 84.62% and 85.53%, respectively. The lesions of anterior pelvis and lip were 80.36%, 75.00% and 78.95%, respectively. The injuries of external upper pelvis and lip were 72.00%, 80.39% and 77.63%, respectively. The injuries of posterior pelvis and lip were 100.00%, 95.71% and 96.05%, respectively. The sensitivity, specificity and accuracy of single hip MRI in the diagnosis of parenchymatous tear of acetabulum labrum were 100.00%, 94.83% and 96.55%, respectively, which was in good agreement with that of hip arthroscopy (Kappa=0.924).Conclusions:3.0T MRI scan of single hip can effectively diagnose the injury of acetabular pelvis and lip, and has high diagnostic value in the differential diagnosis of different parts and types of acetabular pelvis and lip injury.