Objective To investigate the expression of carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) in esophageal squamous cell carcinoma (ESCC) and analyze its correlation with immune cell infiltration and patient prognosis. Methods Three ESCC datasets (GSE161533, GSE26886, and GSE23400) from the GEO database were analyzed to identify differentially expressed genes. CEACAM6 was identified as a key gene through survival analysis. Its expression, prognostic value, and relationship with immune cell infiltration were further explored using databases, such as TIMER. Tissue samples were collected from 162 patients with ESCC. Immunohistochemistry was performed to detect the expression of CEACAM6, immune cell markers (CD4, CD8, CD20, and CD56), and immune checkpoint molecules (HHLA2 and CD40LG). Correlations between CEACAM6 expression and clinicopathological features, immune cell infiltration, and immune checkpoints were analyzed. Results Bioinformatic analysis and clinical sample validation confirmed that CEACAM6 expression was significantly upregulated in ESCC tissues compared with adjacent nontumor tissues (P<0.05). High CEACAM6 expression was closely associated with advanced clinical stage (AJCC Ⅲ-Ⅳ), high T stage (T3-T4), lymph node metastasis, nonulcerative type, and poor prognosis. Furthermore, CEACAM6 expression levels were positively correlated with the infiltration density of CD8⁺ T cells, CD4⁺ T cells, and CD20⁺ B cells within the tumor microenvironment and with the expression of the immune checkpoint molecules HHLA2 and CD40LG (all P<0.05). Conclusion CEACAM6 serves as an independent poor prognostic factor for ESCC. Its high expression is implicated in the modulation of the tumor immune microenvironment by correlating with specific immune cell infiltration and immune checkpoint molecules, suggesting its potential as a novel prognostic biomarker and immunotherapeutic target for ESCC.

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.

Biosensors based on acetylcholinesterase (AChE) are crucial for early diagnosis, less invasive treatment, and drug evaluation of Alzheimer's disease (AD). However, existing technologies often suffer from enzyme conformational changes, leading to altered activity and loss and reduced sensor efficacy. To address this challenge, we developed a novel right-side-out-oriented red blood cell membrane-coated electrochemical biosensors (ROCMCBs) to evaluate AChE inhibitors from traditional Chinese medicines (TCMs) as potential anti-AD agents. The developed right-side-out-oriented coating based on immunoaffinity not only fully exposed the binding sites of AChE on the cell membrane but also ensured its conformation and stability as a peripheral membrane-anchoring protein, which was conducive to maintaining its biological activity and producing optimal interaction with drugs. At the same time, the biosensors exhibited a satisfactory sensitivity (limit of detection = 0.41 pmol/L). Ultimately, six potentially active compounds against AD (baicalin, geniposide, gastrodin, berberine, rhynchophylline, and senkyunolide A) were rapidly identified and evaluated from TCMs. This project provides a promising strategy for developing cell membrane-coated electrochemical biosensors. The application of cell membrane-coated electrochemical biosensors with well-defined cell membrane orientation further expands new perspectives and methods for AChE-targeted anti-AD research.

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 explore the relationship between maternal rejection and internet addiction in college students, as well as the chain mediating role of rejection sensitivity and adaptability.Methods:From March to May 2024, a total of 1 119 college students were surveyed using the short-form Egna Minnen av Barndoms Uppforstran for Chinese(s-EMBU-C), internet addiction test(IAT), rejection sensitivity questionnaire(RSQ), and the China college student adjustment scale(CCSAS).SPSS 26.0 statistical software was used for independent sample t-test, one-way ANOVA, Pearson product moment correlation, and multiple linear regression analysis, and PROCESS 4.0 macro program was used for chain mediation analysis. Results:(1)Maternal rejection (11.19±2.97) was positively correlated with internet addiction (44.89±9.74)( r=0.60, P<0.01) and rejection sensitivity (102.93±55.63)( r=0.63, P<0.01), while negatively correlated with adaptability (200.19±14.18)( r=-0.56, P<0.01) among college students. Rejection sensitivity was positively correlated with internet addiction ( r=0.75, P<0.01) and negatively correlated with adaptability ( r=-0.76, P<0.01). Adaptability was negatively correlated with internet addiction ( r=-0.68, P<0.01). (2)Maternal rejection had a significant direct effect on internet addiction among college students (effect value=0.193, 95% CI=0.145-0.241), accounting for 32.06%(0.193/0.602) of the total effect. Rejection sensitivity mediated the relationship between maternal rejection and internet addiction (effect value=0.290, 95% CI=0.232-0.357), accounting for 48.17%(0.290/0.602) of the total effect. Adaptability also mediated this relationship (effect value=0.028, 95% CI=0.009-0.053), accounting for 4.65%(0.028/0.602) of the total effect. Additionally, there was a chain mediation effect of rejection sensitivity and adaptability on the relationship between maternal rejection and internet addiction (effect value=0.091, 95% CI=0.052-0.130), accounting for 15.12%(0.091/0.602) of the total effect. Conclusion:Maternal rejection can directly influence internet addiction in college students, and it can also indirectly influence internet addiction through the independent mediating effects of rejection sensitivity and adaptability, as well as through the chain mediating effects of both rejection sensitivity and adaptability.

Objective:To explore the relationship between maternal rejection and internet addiction in college students, as well as the chain mediating role of rejection sensitivity and adaptability.Methods:From March to May 2024, a total of 1 119 college students were surveyed using the short-form Egna Minnen av Barndoms Uppforstran for Chinese(s-EMBU-C), internet addiction test(IAT), rejection sensitivity questionnaire(RSQ), and the China college student adjustment scale(CCSAS).SPSS 26.0 statistical software was used for independent sample t-test, one-way ANOVA, Pearson product moment correlation, and multiple linear regression analysis, and PROCESS 4.0 macro program was used for chain mediation analysis. Results:(1)Maternal rejection (11.19±2.97) was positively correlated with internet addiction (44.89±9.74)( r=0.60, P<0.01) and rejection sensitivity (102.93±55.63)( r=0.63, P<0.01), while negatively correlated with adaptability (200.19±14.18)( r=-0.56, P<0.01) among college students. Rejection sensitivity was positively correlated with internet addiction ( r=0.75, P<0.01) and negatively correlated with adaptability ( r=-0.76, P<0.01). Adaptability was negatively correlated with internet addiction ( r=-0.68, P<0.01). (2)Maternal rejection had a significant direct effect on internet addiction among college students (effect value=0.193, 95% CI=0.145-0.241), accounting for 32.06%(0.193/0.602) of the total effect. Rejection sensitivity mediated the relationship between maternal rejection and internet addiction (effect value=0.290, 95% CI=0.232-0.357), accounting for 48.17%(0.290/0.602) of the total effect. Adaptability also mediated this relationship (effect value=0.028, 95% CI=0.009-0.053), accounting for 4.65%(0.028/0.602) of the total effect. Additionally, there was a chain mediation effect of rejection sensitivity and adaptability on the relationship between maternal rejection and internet addiction (effect value=0.091, 95% CI=0.052-0.130), accounting for 15.12%(0.091/0.602) of the total effect. Conclusion:Maternal rejection can directly influence internet addiction in college students, and it can also indirectly influence internet addiction through the independent mediating effects of rejection sensitivity and adaptability, as well as through the chain mediating effects of both rejection sensitivity and adaptability.

Background: This study was performed to investigate the imaging features of mDIXON-Quant sequence (Philips Healthcare) and proton magnetic resonance spectroscopy (1H-MRS) of thigh muscles in patients with stroke-related sarcopenia (SRS). Methods: This prospective case-control study was conducted in 40 patients with SRS, 40 patients without sarcopenia who had a stroke, and 40 healthy volunteers using mDIXON-Quant and 1H-MRS scanning. Skeletal muscle cross-sectional area (CSA) and fat fraction (FF) were analyzed. Results: The skeletal muscle FF value was significantly higher (p<0.05) in patients with SRS and on the affected side. The extracellular fat content of the rectus femoris muscle in normal controls was 4× to 10× the intracellular fat content. A significant increase (p<0.05) in intra- and extracellular fat content was detected in the SRS group. The degree of fat content increase in the SRS group was significantly lower (p<0.05) for extracellular fat than intracellular fat, with a ratio of extracellular to intracellular fat content of <4. The intracellular fat content was significantly higher (p<0.05) in the SRS group. A moderate-to-strong positive correlation existed between intracellular fat content (area 1) and muscle fat percentage. The degree of decrease in CSA in the posterior muscle group was significantly greater (p<0.05). Conclusion Thigh muscle CSA significantly decreased in SRS, while FF increased. The intra- and extracellular fat content of the skeletal muscle was significantly increased, especially the intracellular fat content. SRS was confirmed when the ratio of extracellular fat content to intracellular fat content was <4.

Ultrasound medicine plays a great role in innovative diagnosis and treatment especially when the interdisciplinary characteristics are considered.Cultivating versatile postgraduate students with interdisciplinary capacity is an essential requirement for talent development and disciplinary advancement in ultrasonography.However, limited by factors such as the knowledge structure of the students, the subject background of the supervisors and tutors, policies and regulations, etc., and it is difficult for a single-disciplinary training team to meet the multi-disciplinary requirements of the ultrasound medical profession.In the process of cultivating postgraduate students in cooperation with molecular biology supervisors and tutors, our team has initially explored a set of training paths for inter-disciplinary ultrasound medical postgraduate students.This article systematically sorted out the practical experience of the above-mentioned training model from aspects such as student selection, supervisor and tutor team building, curriculum setting, literature reading ability cultivation, process management, scientific research results sharing mechanism, etc., hoping to provide reference for inter-disciplinary team cooperation in cultivating inter-disciplinary talents.

Purpose To investigate the challenges associ-ated with HER2-low in breast cancer across hospitals of varying regions and grades in China.Drawing on the findings from this investigation,this study will integrate relevant clinical guide-lines,consensus statements,and literature advancements from both domestic and international sources to offer insights for de-veloping guidelines for HER2 detection in Chinese breast cancer cases.Methods The questionnaire implementation method was Tencent WeChat as the carrier and online survey in the form of electronic questionnaire.Results According to the annual HER2 testing results of various medical institutions,compared with HER2 immunohistochemistry(IHC)scores of 3+,HER2 IHC scores of 1+or 2+significantly increased in the proportion range of over 20％(P＜0.001).A total of 854 pathologists provided effective feedback for this survey,55.6％reported dif-ficulties in distinguishing between a score of 0 and 1+in certain cases,primarily due to issues related to staining percentage(83.4％)and the absence of an objective standard for weak staining(78.3％).Regarding testing procedures,417(50.5％)medical institutions used only HER2 IHC 3+as an external control,whereas 139(16.8％)set up a gradient of ex-ternal controls for HER2 IHC scores of 0,1+,2+,and 3+.68.5％of pathologists routinely adhered to the microscope mag-nification rule to assist in HER2 scoring.51.9％of pathologists support a three-category reporting model for HER2 status(nega-tive/low/positive).In cases exhibiting heterogeneity,56.3％of pathologists would accept reporting the percentages of different HER2 scores(0/1+/2+/3+).Conclusion The proportion of breast cancer patients with HER2-low is relatively high in China.Currently,there is no consensus on quality control stand-ards for the pathological diagnosis and testing procedures for HER2-low.Key diagnostic challenges include defining the lower limit of interpretation standards,standardizing testing proce-dures,establishing external controls,refining interpretation methods,and standardizing reporting practices.Given the publi-cation of the DESTINY-Breast06 study results during the revision period,there is an urgent need to develop a new edition of the Chinese guidelines for HER2 testing in breast cancer to provide clearer and more specific strategies.

Purpose To examine the contemporary state of pathological diagnosis for granulomatous lobular mastitis(GLM)in China.Methods Pathologists from across the nation were invited to engage in an online questionnaire survey,which ad-dressed pertinent topics such as the frequency of GLM diagnosis cases,the modalities of pathology reporting,and advancements in etiological research.The collected data were analyzed utili-zing the x2 test or Fisher's exact test.Results A survey of 702 pathologists included 26.9％(189/702)chief physicians,43.3％(304/702)deputy chief physician,26.1％(183/702)attending physicians,and 3.7％(26/702)residents.Partici-pants were from tertiary general hospitals(77.6％,545/702),traditional Chinese medicine hospitals(7.7％,54/702),ma-ternal and child specialty hospitals(6.1％,43/702),and on-cology specialty hospitals(4.0％,28/702).In 2023,there were over 45,000 mastitis diagnoses.A significantly higher per-centage of chief physicians(60.8％,115/189)and deputy chief physicians(52.6％,160/304)could diagnose specific mastitis types compared to attending and resident physicians(x2=23.537,P＜0.001),with"GLM"being the most common diagnosis(55.1％,81/147).In diagnostic work,78.06％(548/702)of pathologists consider patient clinical information,19.2％(135/702)screen specimens for pathogens using special staining,only 6.0％(42/702)of pathologists indicated that their hospitals routinely perform tissue culture for pathogens,with corynebacterium being the most common.Additionally,11.4％(80/702)reported that clinicians regularly test serum sex hormones,most frequently for elevated prolactin,while 7.1％(50/702)check autoimmune status,frequently detecting antibodies without clinical symptoms.Chief physicians had a significantly better understanding of the etiology of GLM com-pared to other physicians(x2=11.969,P=0.003;P=0.007).A majority of pathologists indicated a lack of guidelines for GLM pathology diagnosis(49.4％,347/702)and report standards(76.4％,536/702).Nearly all(97.4％)agreed on the need to establish a consensus for GLM diagnosis.Conclu-sion GLM pathological diagnosis is currently inconsistent and lacks standardization,highlighting the urgent need for a guiding consensus for pathologists.