Objective To investigate the expression pattern and clinical significance of microtubule-associated protein 4(MAP4)in pancreatic adenocarcinoma(PAAD).Methods The immunofluorescence technique was used to detect the expression level of MAP4 in the tissues from 60 PAAD patients visited Liaocheng People's Hospital.The expression levels of MAP4 mRNA and their correlations with patients'prognosis were analyzed using the transcriptome data from 179 PAAD and 332 normal pancreatic tissues in the TCGA and GTEx databases.The correlations of the expression levels of MAP4 with tumor stemness indices,tumor-infiltrating immune cells,and immune checkpoints were analyzed by the bioinformatics method.The small interfering RNA technology was used to silence the expres-sion of MAP4 in PAAD cell lines such as MiaPaCa-2 and PANC-1,and its effects on cell proliferation,migration,and invasion abili-ties were evaluated by the CCK-8 and Transwell assays.Results The results of immunofluorescence showed that the expression levels of MAP4 in PAAD tissues were significantly higher than that in normal pancreatic tissues(P＜0.05).The bioinformatics analysis indi-cated that the expression levels of MAP4 mRNA in PAAD tissues were significantly up-regulated(P＜0.05),and that their high expres-sion was significantly correlated with shorter disease-free and progression-free intervals in patients(P＜0.05).The expression levels of MAP4 were significantly negatively correlated with RNA-related tumor stemness indices(r=-0.55,P＜0.05),but significantly posi-tively correlated with macrophage and dendritic cell infiltration(r＞0.5,P＜0.05).In vitro experimental results confirmed that silencing MAP4 could significantly inhibit the proliferation,migration,and invasion abilities of PAAD cells(P＜0.05).Conclusion MAP4 is highly expressed in PAAD tissues,and its expression levels are closely related to patients'prognosis,tumor stemness,and immune cell infiltration.MAP4 may participate in the progression of PAAD by regulating the biological behavior and immune microenvironment of tumor cells,which provides a new potential target for the diagnosis and treatment of PAAD.

Objective To investigate the expression pattern and clinical significance of microtubule-associated protein 4(MAP4)in pancreatic adenocarcinoma(PAAD).Methods The immunofluorescence technique was used to detect the expression level of MAP4 in the tissues from 60 PAAD patients visited Liaocheng People's Hospital.The expression levels of MAP4 mRNA and their correlations with patients'prognosis were analyzed using the transcriptome data from 179 PAAD and 332 normal pancreatic tissues in the TCGA and GTEx databases.The correlations of the expression levels of MAP4 with tumor stemness indices,tumor-infiltrating immune cells,and immune checkpoints were analyzed by the bioinformatics method.The small interfering RNA technology was used to silence the expres-sion of MAP4 in PAAD cell lines such as MiaPaCa-2 and PANC-1,and its effects on cell proliferation,migration,and invasion abili-ties were evaluated by the CCK-8 and Transwell assays.Results The results of immunofluorescence showed that the expression levels of MAP4 in PAAD tissues were significantly higher than that in normal pancreatic tissues(P＜0.05).The bioinformatics analysis indi-cated that the expression levels of MAP4 mRNA in PAAD tissues were significantly up-regulated(P＜0.05),and that their high expres-sion was significantly correlated with shorter disease-free and progression-free intervals in patients(P＜0.05).The expression levels of MAP4 were significantly negatively correlated with RNA-related tumor stemness indices(r=-0.55,P＜0.05),but significantly posi-tively correlated with macrophage and dendritic cell infiltration(r＞0.5,P＜0.05).In vitro experimental results confirmed that silencing MAP4 could significantly inhibit the proliferation,migration,and invasion abilities of PAAD cells(P＜0.05).Conclusion MAP4 is highly expressed in PAAD tissues,and its expression levels are closely related to patients'prognosis,tumor stemness,and immune cell infiltration.MAP4 may participate in the progression of PAAD by regulating the biological behavior and immune microenvironment of tumor cells,which provides a new potential target for the diagnosis and treatment of PAAD.

Objective To investigate the governance requirements for real world data(RWD)in China's medical standards and specifications,summarize key technical aspects of data governance,and provide refer-ences for RWD governance-related research.Methods Computerized searches were conducted in CNKI,Wan-fang Data,VIP,and SinoMed,as well as the official websites of 29 national medical societies and the National Standard Information Public Service Platform,covering all records from inception to December 12,2023.A data extraction form was developed,and the included standards and specifications were categorized according to the first six RWD governance processes outlined in the Guidelines for Real World Data Used to Generate Real-World Evidence(Trial)issued by the National Medical Products Administration:data security,data extraction,data cleaning,data transformation,data transmission and storage,and quality control.Relevant content on data governance was systematically summarized and comparatively analyzed.Results A total of 32 standards and specifications were included,comprising 15 foundational medical data standards and 17 data gov-ernance technical specifications.Among these,6 addressed data security,6 covered data extraction,5 focused on data cleaning,5 involved data transformation,6 pertained to data transmission and storage,and 4 discussed quality control.Foundational medical data standards included data description elements,terminology,and format standards,broadly covering essential data elements and meeting basic standardization needs.Data gov-ernance technical specifications primarily provided general guidelines for medical data governance,emphasizing requirements and recommendations.While requirements for data security and extraction were relatively well-de-fined,technical guidance on data transformation and quality control remained limited,and implementation pathways for data cleaning,transmission,and storage were insufficiently detailed.Conclusions As real-world evidence plays an increasingly critical role in healthcare decision-making,China's medical standards and speci-fications have established a preliminary governance framework for RWD.However,technical details and practi-cal implementation of RWD governance still require further refinement.

Objective To investigate the governance requirements for real world data(RWD)in China's medical standards and specifications,summarize key technical aspects of data governance,and provide refer-ences for RWD governance-related research.Methods Computerized searches were conducted in CNKI,Wan-fang Data,VIP,and SinoMed,as well as the official websites of 29 national medical societies and the National Standard Information Public Service Platform,covering all records from inception to December 12,2023.A data extraction form was developed,and the included standards and specifications were categorized according to the first six RWD governance processes outlined in the Guidelines for Real World Data Used to Generate Real-World Evidence(Trial)issued by the National Medical Products Administration:data security,data extraction,data cleaning,data transformation,data transmission and storage,and quality control.Relevant content on data governance was systematically summarized and comparatively analyzed.Results A total of 32 standards and specifications were included,comprising 15 foundational medical data standards and 17 data gov-ernance technical specifications.Among these,6 addressed data security,6 covered data extraction,5 focused on data cleaning,5 involved data transformation,6 pertained to data transmission and storage,and 4 discussed quality control.Foundational medical data standards included data description elements,terminology,and format standards,broadly covering essential data elements and meeting basic standardization needs.Data gov-ernance technical specifications primarily provided general guidelines for medical data governance,emphasizing requirements and recommendations.While requirements for data security and extraction were relatively well-de-fined,technical guidance on data transformation and quality control remained limited,and implementation pathways for data cleaning,transmission,and storage were insufficiently detailed.Conclusions As real-world evidence plays an increasingly critical role in healthcare decision-making,China's medical standards and speci-fications have established a preliminary governance framework for RWD.However,technical details and practi-cal implementation of RWD governance still require further refinement.

Objective To establish a clinical-CT model,and to observe its value for evaluating lymphovascular invasion(LVI)and/or perineural invasion(PNI)in esophageal squamous cell carcinoma(ESCC).Methods Data of 156 ESCC patients were retrospectively analyzed.The patients were divided into positive group(n=58,LVI[+]and/or PNI[+])and negative group(n=98,LVI[-]and PNI[-])according to postoperative pathological results.Clinical and CT data were compared between groups.Logistic regression analysis was performed to establish a model,and its efficacy of evaluating ESCC LVI and/or PNI was analyzed.Results Significant differences of carcinoembryonic antigen(CEA),carbohydrate antigen 199(CA199),tumor thickness,tumor volume and CT venous phase value(CTV),the difference between CTV and CT plain phase value(CTP)(△CTV-P)and venous phase enhancement rate(V％)were found between groups(all P＜0.05),and the area under the curve(AUC)of the above parameters for evaluating ESCC LVI and/or PNI was 0.702,0.690,0.731,0.744,0.621,0.631 and 0.599,respectively.CEA,CA199,tumor thickness,tumor volume and CTV were all independent predictive factors for ESCC LVI and/or PNI.A combined model was established based on the above features,and its accuracy,sensitivity and specificity for evaluating ESCC LVI and/or PNI was 82.05％,65.52％and 91.84％,respectively,with AUC of 0.838,higher than that of each single parameter(all P＜0.05).Conclusion The established clinical-CT model could effectively evaluate ESCC LVI and/or PNI.

Objective To observe the value of intratumoral and peritumoral CT radiomics for evaluating KRAS gene status in patients with colorectal adenocarcinoma.Methods Totally 245 patients with colorectal adenocarcinoma were retrospectively enrolled and divided into mutant group(n=139)and wild group(n=106)according to KRAS gene status,also divided into training set(n=171)and test set(n=74)at a ratio of 7∶3.Clinical data were compared between groups,and clinical factors were screened with logistic regression analysis to establish a clinical model.Based on enhanced venous phase CT images,intratumoral volume of interest(VOI),peritumoral VOI,and intratumoral+peritumoral VOI were delineated,radiomics features were extracted,and radiomics models were constructed.The combination model was constructed based on the best radiomics model combined with clinical factors.The value of each model for evaluating KRAS gene status in patients with colorectal adenocarcinoma was analyzed.Results Significant differences of patients’gender and carcinoembryonic antigen(CEA)were found between mutant group and wild group(both P＜0.05),which were independent impact factors of KRAS gene status in patients with colorectal adenocarcinoma(both P＜0.05).The area under the curve(AUC)of clinical model for evaluating KRAS gene status in patients with colorectal adenocarcinoma in training set and test set was 0.633 and 0.658,respectively.Intratumoral+peritumoral 3 mm model was the best radiomics model,with AUC of 0.921 and 0.894 in training set and test set,respectively.AUC of the combination model in training set and test set was 0.949 and 0.956,respectively.In training set,significant differences of AUC were found between clinical model and intratumoral+peritumoral 3 mm model,also between clinical model and combination model(both P＜0.001),while in test set,significant differences of AUC were found between each two models(all P＜0.05).Conclusion Intratumoral+peritumoral 3 mm radiomics based on enhanced venous phase CT could help to evaluate KRAS gene status in patients with colorectal adenocarcinoma.Combining with patients’gender and CEA could further improve efficacy of this model.

Objective To observe the value of contrast-enhanced CT radiomics combined with clinical and hematology indicators for predicting lymph node(LN)metastasis(LNM)of esophageal squamous cell carcinoma(ESCC).Methods Totally 218 ESCC patients were retrospectively enrolled.Stage pN1 and pN2 were clustering as LNM(n=90),while stage pN0 were taken as non-LNM(n=128).The patients were divided into training set(n=174)and test set(n=44)at the ratio of 8∶2.In training set,clinical and LN imaging features which could be used to independently judge LNM were screened and a clinical-imaging model was constructed.The hematological indicators that might be associated with ESCC LNM were screened,and a hematological model was constructed.Radiomics features in LN ROI and ESCC volume of interest(VOI)were extracted based on venous-phase contrast-enhanced CT images,and those might be associated with LNM were screened,and a radiomics model was constructed.Finally a combined model was constructed based on all the above features.The efficacy of each model for diagnosing LNM was evaluated with the area under the curve(AUC)of receiver operating characteristic curves,and the clinical net benefit was evaluated using decision curve analysis(DCA).Results Body mass index(BMI)and internal necrosis of target LN were both independent judging factors for ESCC LNM(both P＜0.05),and AUC of clinical-imaging model for diagnosing LNM in training and test sets was 0.747 and 0.687,respectively.Seven hematological indicators were included in hematological model,and AUC in training and test sets was 0.623 and 0.583,respectively.Ten LN radiomics features and 15 ESCC radiomics features were included in radiomics model,and AUC in training and test sets was 0.769 and 0.745,respectively.AUC of the combined model for diagnosing LNM in training and test sets was 0.822 and 0.739,respectively,better than other models in training set(all P＜0.05),but no significantly different in test set(all P＞0.05).DCA showed that combined model had higher net gain than the other models in 0.55-0.80 threshold probability interval.Conclusion Combined model based on venous-phase contrast-enhanced CT radiomics and clinical and hematology indicators could relatively effectively evaluate ESCC LNM,which might bring some promotions in clinical benefit.

Although DNA 5-hydroxymethylcytosine (5hmC) is recognized as an important epige-netic mark in cancer, its precise role in lymph node metastasis remains elusive. In this study, we investigated how 5hmC associates with lymph node metastasis in breast cancer. Accompanying with high expression of TET1 and TET2 proteins, large numbers of genes in the metastasis-positive pri-mary tumors exhibit higher 5hmC levels than those in the metastasis-negative primary tumors. In contrast, the TET protein expression and DNA 5hmC decrease significantly within the metastatic lesions in the lymph nodes compared to those in their matched primary tumors. Through genome-wide analysis of 8 sets of primary tumors, we identified 100 high-confidence metastasis-associated 5hmC signatures, and it is found that increased levels of DNA 5hmC and gene expression of MAP7D1 associate with high risk of lymph node metastasis. Furthermore, we demonstrate that MAP7D1, regulated by TET1, promotes tumor growth and metastasis. In conclusion, the dynamic 5hmC profiles during lymph node metastasis suggest a link between DNA 5hmC and lymph node metastasis. Meanwhile, the role of MAP7D1 in breast cancer progression suggests that the metastasis-associated 5hmC signatures are potential biomarkers to predict the risk for lymph node metastasis, which may serve as diagnostic and therapeutic targets for metastatic breast cancer.

Gegen Qinlian Decoction (GQD), a traditional Chinese medicine (TCM) formula, has long been used for the treatment of common metabolic diseases, including type 2 diabetes mellitus. However, the main limitation of its wider application is ingredient complexity of this formula. Thus, it is critically important to identify the major active ingredients of GQD and to illustrate mecha-nisms underlying its action. Here, we compared the effects of GQD and berberine, a hypothetical key active pharmaceutical ingredient of GQD, on a diabetic rat model by comprehensive analyses of gut microbiota, short-chain fatty acids, proinflammatory cytokines, and ileum transcriptomics. Our results show that berberine and GQD had similar effects on lowering blood glucose levels, modulating gut microbiota, inducing ileal gene expression, as well as relieving systemic and local inflammation. As expected, both berberine and GQD treatment significantly altered the overall gut microbiota structure and enriched many butyrate-producing bacteria, including Faecalibacterium and Roseburia, thereby attenuating intestinal inflammation and lowering glucose. Levels of short-chain fatty acids in rat feces were also significantly elevated after treatment with ber-berine or GQD. Moreover, concentration of serum proinflammatory cytokines and expression of immune-related genes, including Nfkb1, Stat1, and Ifnrg1, in pancreatic islets were significantly reduced after treatment. Our study demonstrates that the main effects of GQD can be attributed to berberine via modulating gut microbiota. The strategy employed would facilitate further stan-dardization and widespread application of TCM in many diseases.

In this study, a novel tetrapeptide hydrophilic interaction chromatography (HILIC) material was synthesized, and corresponding enrichment method for glycopeptides was developed.The tetrapeptide modified silica gel materials (denoted as Poly-DAPD) were synthesized by atom-transfer radical-polymerization (ATRP) and characterized by N2 adsorption desorption and thermometer, thermal gravimetric analyzer (TGA) and X-ray photoelectron spectrometer (XPS).The characterization results indicated that tetrapeptide had been successfully synthesized on silica gel.Poly-DAPD materials showed high enrichment selectivity toward fetuin glycopeptides under solid-phase extraction (SPE) mode.Comparing with commercialized ZIC-HILIC in glycopeptides enrichment of fetuin digest which mixed with 5 mole ratio of albumin bovine (BSA), the as-prepared materials showed higher selectivity in both aspects of the identified number of glycopeptides and anti-interference property.The SPE results demonstrated that the tetrapeptide-based HILIC materials could be a potential tool in large-scale glycosylation analysis.