Background:Colorectal cancer(CRC)is a major cause of cancer death worldwide,and about 30%to 35%of CRC arises from the serrated pathway.Aims:To analyze the clinicopathological and histomorphological characteristics of colorectal superficially serrated adenoma(SuSA),and to investigate the malignant transformation potential of SuSA and further elucidate the role of RSPO2 and RSPO3 in its carcinogenic mechanism.Methods:A total of 169 serrated colorectal lesions confirmed pathologically and fulfilling the inclusion criteria were collected from Shanxi Bethune Hospital from February 2019 to May 2023.Among them,there were 84 cases of SuSA,23 sessile serrated lesions(SSL),32 traditional serrated adenomas(TSA),and 30 hyperplastic polyps(HP).Thirty-nine cases of tubular adenoma(TA),32 CRC,and 33 normal colorectal mucosal tissues were served as controls.The clinicopathological and histomorphological parameters were collected and recorded.Immunohistochemical staining was used to detect the protein expressions of RSPO2,RSPO3,p53,MLH1 and β-catenin.Results:Patients with SuSA were male predominance,with a mean age of 51.89 years.The average diameter of the lesions was 0.20 cm,predominantly located in the left colon and rectum,and frequently complicated with other polyps/adenomas.The histomorphological features of SuSA were as follows:the glandular necks and crypt bases exhibited straight tubular structure with low-grade dysplasia;the superficial layer demonstrated a serrated architecture,with or without dysplasia,and contained a variable number of goblet cells.Immunohistochemically,there were no statistically significant differences in RSPO2,RSPO3,and p53 expressions between SuSA and TSA(all P>0.05).A strong positive correlation was observed between RSPO2 and RSPO3 in colorectal lesions,excluding HP.Expressions of MLH1 and β-catenin showed no statistically significant differences between SuSA and other colorectal lesions(all P>0.05).Conclusions:SuSAs are more common in males,occur mostly in the left colon and rectum,and are often associated with other polyps/adenomas.They might be precursors of KRAS-mutated TSA and microsatellite stable CRC with high malignant potential.RSPO2 and RSPO3 might play an important role in the carcinogenesis of SuSA.

Objective To investigate whether ginsenoside Rg3 can ameliorate glomerular endothelial injury in diabetic nephropathy(DN)mice through Ras homologous gene family member A(Rho A)/Rho-associated coiled-coil forming protein kinase,(ROCK1)/NLR family pyrin domain protein 3(NLRP3)pathway.Methods Forty mice were randomly divided into 4 groups:control group,DN group,ginsenoside(ginsenoside Rg3)group and RhoA/ROCK pathway inhibition(FD)group,with 10 mice in each group.Fasting blood glucose(FBG)was measured by glucose meter.The levels of urinary protein,urea nitrogen(BUN)and serum creatinine(SCr)were detected by ELISA.PAS staining was used to detect glomerular morphology and structure and to evaluate glomerular injury index(GDI).The expression of platelet-endothelial cell adhesion molecule(PECAM-1 or CD31),von Willefibrilia factor(vWF),RhoA,ROCK and NLRP3 protein related to pyrodeath were detected by immunofluorescence staining.Western blotting detected the expression of intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),the inflammatory factor interleukin-1β(IL-1β)and IL-18 protein in the glomerulus.Results Compared with the control group,the levels of FPG,urinary protein,BUN and SCr in DN group were increased,and the differences were statistically significant(t=17.59～43.81,all P<0.05).The glomerular structure was significantly damaged and GDI was increased(t=20.73,P<0.05).The expressions of CD31,RhoA,ROCK and NLRP3 in glomeruli were increased,while the expression of vWF was decreased.The expressions of ICAM-1,VCAM-1,IL-1β and IL-18 in renal tissues were increased,and the differences were statistically significant(t=27.95～40.10,all P<0.05).Compared with the DN group,the levels of FPG,urinary protein,BUN and SCr in ginsenoside group were decreased,and the differences were statistically significant(t=14.87～20.33,all P<0.05).The damage of glomerular structure was improved and GDI was decreased(t=19.80,P<0.05),the expression of CD31,RhoA,ROCK and NLRP3 in glomerular was decreased,and the expression of vWF was increased.The expressions of ICAM-1,VCAM-1,IL-1β and IL-18 in renal tissues of FD group were decreased,and the differences were statistically significant(t=12.62～39.68,all P<0.05).Conclusion Ginsenosides Rg3 can improve the level of glomerular endothelial injury and pyroptosis in DN mice by down-regulating RhoA/ROCK/NLRP3 pathway.

Objective To assess the nutritional status of patients undergoing preventive ileostomy and identify the factors that influence the status,thereby providing guidance for medical staff to formulate nutritional intervention strategies.Methods Convenient sampling was used to select 239 patients undergoing preventive ileostomy who were attending follow-up visits or hospitalized while awaiting stoma closure at 4 tertiary hospitals in Guangzhou from November 2023 to July 2024.Data was collected by a general information questionnaire and the Patient-Generated Subjective Global Assessment tool.Factors influencing nutritional status were analyzed by multivariable ordinal Logistic regression.Results Of the 239 patients,227 provided valid responses.The nutritional status was categorized as follows:64 patients(28.19％)had good nutrition or were at risk of malnutrition;104(45.81％)exhibited moderate malnutrition;59(25.99％)had severe malnutrition.Multivariable ordinal Logistic regression analysis revealed that preoperative cumulative chemotherapy cycles,postoperative duration,age,current dietaiy type,use of oral nutritional supplements(ONS),enterostomy-related complications,and stoma self-care ability were significant predictors of nutritional status(P＜0.05).Conclusion Patients undergoing preventive ileostomy are at a high risk of moderate to severe malnutrition and ONS provided to patients did not result in a meaningful improvement in their nutritional health.Specifically,those patients with a higher number of cumulative preoperative chemotherapy cycles,shorter postoperative recovery time,aged ≥ 65 years,liquid diet or semi-liquid diet,experiencing enterostomy-related complications,or poor stoma self-care ability,are particularly vulnerable to malnutrition.These findings underscore the need for medical staff to formulate intervention strategies based on these factors to improve nutritional status of patients undergoing preventive ileostomy.

Objective:To evaluate the role of the TANK-binding kinase 1 (TBK1)/receptor-interacting protein kinase 1 (RIPK1) signaling pathway in postoperative cognitive dysfunction (POCD) in aged mice.Methods:Fifty SPF healthy male C57BL/6 mice, aged 18 months, weighing 20-25 g, were divided into 5 groups ( n=10 each) using a table of random numbers: control group (group C), POCD group, dimethyl sulfoxide group, GSK group and GSK+ Nec-1 group. A mouse model of POCD was established by the closed reduction internal fixation of the left tibial fracture in anesthetized animals. Dimethyl sulfoxide, TBK1 inhibitor GSK8612 and RIPK1 inhibitor Nec-1 (0.5 μl/side) were stereotactically injected into the hippocampal CA1 region at 30 min before operation. Cognitive function was assessed using the contextual fear conditioning test before operation and at 3 days after operation. The mice were then anesthetized and sacrificed, and the hippocampal tissues were obtained for determination of the expression of TBK1, RIPK1, interleukin-lbeta (IL-1β), tumor necrosis factor-alpha (TNF-α), activator protein 1 (AP-1) and Nestin (by Western blot), the expression of Bcl-2, Bax and caspase-3 mRNA (by fluorescent quantitative real-time polymerase chain reaction) and for examination of TBK1/RIPK1 molecular interactions and neural stem cell proliferation in the hippocampal dentate gyrus (DG) region (by immunofluorescent staining). Results:Compared with C group, the percentage of freezing time was significantly decreased at 3 days after operation, the expression of Bax mRNA, caspase-3 mRNA, RIPK1, IL-1β, TNF-α and AP-1 was up-regulated, the expression of TBK1, Bcl-2 mRNA and Nestin was down-regulated, and the proliferation of neural stem cells in the hippocampal DG region was decreased in POCD group ( P<0.05 or 0.01). Compared with POCD group, the percentage of freezing time was significantly decreased at 3 days after operation, the expression of Bax mRNA, caspase-3 mRNA, IL-1β, TNF-α and AP-1 was up-regulated, the expression of TBK1, Bcl-2 mRNA and Nestin was down-regulated, and the proliferation of neural stem cells in the hippocampal DG region was decreased in GSK group ( P<0.05 or 0.01). Compared with GSK group, the percentage of freezing time was significantly increased at 3 days after operation, the expression of Bax mRNA, caspase-3 mRNA, IL-1β, TNF-α and AP-1 was down-regulated, the expression of TBK1, Bcl-2 mRNA and Nestin was up-regulated, and the proliferation of neural stem cells in the hippocampal DG region was increased in GSK+ Nec-1 group ( P<0.05 or 0.01). Conclusions:The TBK1/RIPK1 signaling pathway is involved in the pathogenesis of POCD in aged mice.

Aiming at the classification and lesion localization of giga-pixel pathology whole slide images of breast cancer,a bidirectional gated recurrent unit attention based multi-instance learning(ABMIL-BiGRU)model is proposed for interpretable prediction of H&E stained breast cancer lymph node metastasis images.The method uses two orthogonal bidirectional gated recurrent units to establish the long-short distance dependencies between the features in the row and column directions of the image block,thereby embedding the spatial position and context information of the image block,and then quantifies the attention score of each feature representation through attention multi-instance pooling,thereby achieving whole slide image-level feature aggregation and generating interpretable heat maps.The results show that ABMIL-BiGRU model has an average accuracy of 0.918 6 and an AUCof 0.9467 on the breast cancer metastasis dataset,realizing high-precision prediction of whole slide images and localization of regions of interest,and also providing human-interpretable features at the image block level.The proposed model solves the"accuracy-interpretability trade-off"problem to a certain extent,and its superior performance provides a new paradigm for the clinical application of computer-aided diagnosis and intelligent systems.

Objective To analyze relevant literature on the essence of traditional Chinese medicine syndromes using bibliometric methods;To understand the current research status and hotspots in this field;To provide references for relevant research.Methods Research literature about the essence of TCM syndromes was retrieved from CNKI,VIP,Wanfang Data and CBM from 1st Jan.1979 to 30th June 2024.CiteSpace 6.2 software was used to conduct visualization analysis on authors,institutions,keywords,etc.Results A total of 695 articles were included,with an overall upward trend in publication volume followed by stabilization.The authors who published more articles included Luo Ren(13 articles),Zhao Xiaoshan(13 articles),Li Zegeng(12 articles),etc.The institutions with more publications included Shandong University of Traditional Chinese Medicine(35 articles),Beijing University of Chinese Medicine(33 articles),Chengdu University of Traditional Chinese Medicine(26 articles),etc.Institutions were clustered regionally,and cooperation was mostly between TCM universities and their affiliated hospitals,with less cross regional communication.High frequency keywords included kidney yang deficiency syndrome,metabolomics,animal models,TCM syndrome types,coronary heart disease,lung qi deficiency syndrome,spleen qi deficiency syndrome,liver depression and spleen deficiency syndrome,etc;Keyword clustering covered aspects such as biomolecules and metabolism,TCM syndromes,pathology,disease models,and animal research.Conclusion Research in the field of the essence of TCM syndromes is gradually receiving more attention.Exploring the essence of TCM syndromes at the molecular level through techniques such as genomics,transcriptomics,metabolomics and proteomics is a research hotspot and trend in this field.

Radiotherapy is main method in treating cervical cancer,and the rapid advancement of artificial intelligence(AI)technique is providing entirely new solutions for radiotherapy for cervical cancer.The AI means that is represented by deep learning is deeply integrating into the whole process of diagnosis,treatment and management for cervical cancer,which can promote intelligent and precise development of radiotherapy workflows.Currently,the applied cores of AI in radiotherapy for cervical cancer include image registration,target delineation,optimization of radiotherapy planning and risk assessment,which can significantly enhance efficiency and precision of treatment.But,AI is facing some challenges in clinical applications include data quality,and algorithm's robustness and interpretability at the same time.Depended on the above analyses,this paper systematically reviewed the frontier applications and progress in practice of AI in radiotherapy for cervical cancer,which especially analyzed technical advantages and limitations of AI in key link,and explored its development path and coping strategy in clinical promotion and standard application in future.It is purpose to provide theoretical references for clinical practice of precise and accurate radiotherapy for cervical cancer.

Objective To investigate how vismodegib(Vis)influences the pathogenesis of basal cell carcinoma(BCC)via a chromatin remodeling factor,bromodomain containing protein 9(BRD9),and to analyze the expression profile of BRD9 in BCC and its relationship with the immune checkpoint,programmed cell death-1 ligand 1(PD-L1)and the Hedgehog(Hh)signaling pathway.Methods ① A UVB-induced BCC model was established in SKH-1 hairless background Ptch1+/-;LacZ reporter mice.Then the mice were treated with Vis,and those without treatment served as control.X-gal staining,immunohistochemistry(IHC)staining,immunofluorescence(IF)assay,and Western blotting were used to assess the expression and localization of BRD9 and PD-L1 in tumor tissues and to evaluate immune-cell infiltration.② In vitro,mouse BCC cell line ASZ001(ASZ cells)were treated with Vis or a BRD9 degrader(dBRD9),and BRD9-overexpressing cells were generated.Cell viability and the protein and mRNA levels of BRD9,PD-L1,Gli1,and cyclin D1(Ccnd1)were measured.ChIP-qPCR was performed to examine BRD9 and H3K27ac enrichment at the PD-L1 promoter,including the promoter-proximal site(P1)and an upstream active segment(P2).Results ① At the tissue level,BRD9 was highly expressed in BCC,and co-localization of BRD9 and PD-L1 was observed within tumor regions,with evident immune-cell infiltration.Vis markedly suppressed UVB-induced BCC formation,reduced the probability of large-volume tumors(by probability-density analysis),decreased the X-gal-positive lesion area(P<0.000 1),down-regulated BRD9(P=0.024 9),and attenuated immune-cell infiltration.② At the cellular level,Vis treatment reduced cell viability and down-regulated BRD9,Gli1,and Ccnd1 in ASZ cells(P<0.000 1).dBRD9 inhibited ASZ cell viability in a dose-dependent manner and decreased PD-L1,Gli1,and Ccnd1(P<0.000 1),whereas its overexpression increased the expression of these molecules(P<0.000 1).In ASZ cells,BRD9 and H3K27ac were enriched at the PD-L1 promoter P1/P2 regions.Treatment with dBRD9 or Vis reduced BRD9 and H3K27ac enrichment at P1/P2 regions(P<0.000 1).Conclusion In BCC,BRD9 maintains chromatin activation at the proximal PD-L1 promoter and modulates Hh/Gli1 signaling,thereby promoting immune evasion.Vis remodels the tumor immune microenvironment by inhibiting the Hh-BRD9-PD-L1 axis.

OBJECTIVE: To explore the genetic etiology of fetal skeletal dysplasia using whole exome sequencing (WES) and copy number variation sequencing (CNV-seq) techniques, and the feasibility of using WES as the first-tier method for such fetuses. METHODS: Seventy four fetuses with skeletal dysplasia detected by prenatal ultrasound at the Genetic Testing Center of the Women and Children's Hospital Affiliated to Qingdao University from January 2020 to August 2024 were selected as the study subjects. Fetal muscle and peripheral blood samples of the pregnant women and their spouses were collected and subjected to WES analysis. CNV-seq was carried out on all fetal muscle tissue samples. And the results were compared with the CNVs indicated by WES. Genetic etiologies were analyzed across different subtypes of skeletal dysplasia. And the feasibility of using WES as the first-tier genetic test for similar fetuses was assessed, in addition with a systematic cost-effectiveness analysis. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: QFELL-YJ-2024-201). RESULTS: A total of 50 fetuses were diagnosed, which yielded a diagnostic rate of 67.57%. These included 6 chromosomal aneuploidies, 4 chromosomal CNVs and 40 monogenic disorders. The monogenic diseases had involved 46 variant sites in 23 pathogenic genes, among which 12 were unreported previously, including MYH3: c.735T>C, ALPL: c.1324C>T, NEK9: c.1973G>A, MAGEL2: c.2024_2025del, LMBR1: c.423+4914A>C, NEB: c.21273_21276del, COL1A1: c.2651G>C and c.2758G>C, ASPM: c.2473delinsGA, TBX5: c.704G>A, DYNC2H1: c.10893del, and DYNC2I2: c.1270C>T. Substantial concordance was reached between WES-derived CNV calls and CNV-seq findings. No clinically significant CNV was exclusively detected by CNV-seq. Cost-effectiveness modeling demonstrated that implementing WES as the first-tier genetic testing method could reduce the total expenditure when WES unit cost remained below 6.4 folds that of the CNV-seq. CONCLUSION Genetic variants including single nucleotide variations (SNV) of monogenic disorders, chromosomal aneuploidies and genomic CNVs are important causes for fetal skeletal dysplasia. WES is an accurate and efficient method for analyzing the etiology of fetal skeletal dysplasia, particularly in those with a family history of similar phenotype or maternal history of adverse pregnancies.
Humans ; Exome Sequencing/methods* ; Female ; Pregnancy ; DNA Copy Number Variations/genetics* ; Genetic Testing/methods* ; Prenatal Diagnosis/methods* ; Adult ; Male ; Fetus ; Bone Diseases, Developmental/diagnosis* ; Ultrasonography, Prenatal

Radiotherapy is main method in treating cervical cancer,and the rapid advancement of artificial intelligence(AI)technique is providing entirely new solutions for radiotherapy for cervical cancer.The AI means that is represented by deep learning is deeply integrating into the whole process of diagnosis,treatment and management for cervical cancer,which can promote intelligent and precise development of radiotherapy workflows.Currently,the applied cores of AI in radiotherapy for cervical cancer include image registration,target delineation,optimization of radiotherapy planning and risk assessment,which can significantly enhance efficiency and precision of treatment.But,AI is facing some challenges in clinical applications include data quality,and algorithm's robustness and interpretability at the same time.Depended on the above analyses,this paper systematically reviewed the frontier applications and progress in practice of AI in radiotherapy for cervical cancer,which especially analyzed technical advantages and limitations of AI in key link,and explored its development path and coping strategy in clinical promotion and standard application in future.It is purpose to provide theoretical references for clinical practice of precise and accurate radiotherapy for cervical cancer.