The interaction between m⁶A-methylated RNA and chromatin modification remains largely unknown. We found that targeted inhibition of bromodomain-containing protein 4 (BRD4) by siRNA or its inhibitor (JQ1) significantly decreases mRNA m⁶A levels and suppresses the malignancy of breast cancer (BC) cells via increased expression of demethylase AlkB homolog 5 (ALKBH5). Mechanistically, inhibition of BRD4 increases the mRNA stability of ALKBH5 via enhanced binding between its 3' untranslated regions (3'UTRs) with RNA-binding protein RALY. Further, BRD4 serves as a scaffold for ubiquitin enzymes tripartite motif containing-21 (TRIM21) and ALKBH5, resulting in the ubiquitination and degradation of ALKBH5 protein. JQ1-increased ALKBH5 then demethylates mRNA of extra spindle pole bodies like 1 (ESPL1) and reduces binding between ESPL1 mRNA and m⁶A reader insulin like growth factor 2 mRNA binding protein 3 (IGF2BP3), leading to decay of ESPL1 mRNA. Animal and clinical studies confirm a critical role of BRD4/ALKBH5/ESPL1 pathway in BC progression. Further, our study sheds light on the crosstalks between histone modification and RNA methylation.

Objective:? To summarize the achievements in improving the consistency of clinical liquid chromatography-tandem mass spectrometry (LC-MS/MS) testing results.Methods:? From 2021 to 2024, Zhongshan Hospital Affiliated to Fudan University recruited laboratories voluntarily participating in the MSHP (Clinical LC-MS/MS Testing Consistency Program). As of Batch 202404, a total of 76 laboratories had enrolled, including 60 medical institutions (all tertiary hospitals) and 16 third-party laboratories. Test items were established, and comparative samples were distributed regularly-each item′s samples covered three concentrations (high, medium, and low). Samples were shipped via cold chain and tested within one week. Our laboratory′s measurements served as the target, with participating labs′ results within ±25% of the target deemed qualified. Passing-Bablok regression and Bland-Altman analysis were used to assess consistency.Results:Taking 3-MT (3-methoxytyramine) as an example, the coefficients of variation (CVs) for the project′s three concentration levels improved from 17.00%, 47.18%, and 4.88% in the first comparative batch to 9.59%, 9.59%, and 6.1% in Batch 202404. Passing-Bablok regression results for the 5 units participating in 3-MT testing showed that Laboratory A had proportional bias but no systematic bias (regression slope [95% CI]: 0.903 [0.862-0.952]; intercept [95% CI]: 0.912 [-1.921-6.073]). The remaining laboratories exhibited no proportional or systematic bias with the target (Laboratory B: slope 1.031 [0.961-1.147], intercept-0.733 [-4.641-8.272]; Laboratory C: slope 0.982 [0.940-1.009], intercept-0.576 [-2.675-1.891]; Laboratory D: slope 0.973 [0.939-1.066], intercept-1.168 [-6.108-1.649]; Laboratory E: slope 0.999 [0.905-1.051], intercept-1.876 [-6.111-3.508]). Bland-Altman analysis indicated that all 5 laboratories′ results generally showed good consistency with the target. Through quality feedback and optimizing sample preparation concentrations, result consistency was enhanced.? Conclusion:? Clinical LC-MS/MS testing consistency programs contribute to improving the comparability of test results.

OBJECTIVE To analyze the characteristics of peripheral blood leukocyte morphology,scatterplot and leukocyte cell population data(CPD)in patients with severe fever and thrombocytopenia syndrome(SFTS),infec-tious mononucleosis(IM)and hemorrhagic fever with renal syndrome(HFRS),and to explore their differences.METHODS Blood routine examination data were collected,including microscopic cell images,scatter-plots and leukocyte CPD study parameters from 44 patients diagnosed with SFTS,17 with IM and 11 with HFRS at the First Affiliated Hospital with Nanjing Medical University from Mar.2023 to May 2024.These data were compared with those of healthy controls to explore specific differences in the characteristics of peripheral blood leu-kocyte.RESULTS All 3 viral infections exhibited a left shift in the nucleus under the microscope,yet the blood routine scatterplot and microscopic leukocyte characteristics differed.SFTS commonly presented with a plasmacy-toid reactive lymphocyte with scattered points appearing in the upper reactive lymphocyte area of the scatterplot.IM showed a predominant irregular fried-egg-like reactive lymphocyte under the microscope,with the scatterplot typically presenting a rocket-like shape,in patients with HFRS,there was a significant increase in im-mature granulocytes in peripheral blood and plasmacytoid reactive lymphocyte scatter points with blue immature granulocyte scatter points visible.The fluorescence distribution width(MO-WY)of monocytes in the SFTS group was lower than those in the IM,HFRS and healthy control groups(P＜0.05).Compared to SFTS,IM and healthy control groups,the lateral scattered light distribution width(NE-WX)and fluorescence distribution width(NE-WY)of neutrophils in the HFRS group were both increased(P＜0.05).CONCLUSIONS There are signifi-cant differences in the characteristics of peripheral blood leukocyte morphology,scatterplot and CPD study param-eters among three viral infection diseases SFTS,IM and HFRS.These differences can provide early clinical advice and assist in clinical differentiation.

OBJECTIVE To analyze the characteristics of peripheral blood leukocyte morphology,scatterplot and leukocyte cell population data(CPD)in patients with severe fever and thrombocytopenia syndrome(SFTS),infec-tious mononucleosis(IM)and hemorrhagic fever with renal syndrome(HFRS),and to explore their differences.METHODS Blood routine examination data were collected,including microscopic cell images,scatter-plots and leukocyte CPD study parameters from 44 patients diagnosed with SFTS,17 with IM and 11 with HFRS at the First Affiliated Hospital with Nanjing Medical University from Mar.2023 to May 2024.These data were compared with those of healthy controls to explore specific differences in the characteristics of peripheral blood leu-kocyte.RESULTS All 3 viral infections exhibited a left shift in the nucleus under the microscope,yet the blood routine scatterplot and microscopic leukocyte characteristics differed.SFTS commonly presented with a plasmacy-toid reactive lymphocyte with scattered points appearing in the upper reactive lymphocyte area of the scatterplot.IM showed a predominant irregular fried-egg-like reactive lymphocyte under the microscope,with the scatterplot typically presenting a rocket-like shape,in patients with HFRS,there was a significant increase in im-mature granulocytes in peripheral blood and plasmacytoid reactive lymphocyte scatter points with blue immature granulocyte scatter points visible.The fluorescence distribution width(MO-WY)of monocytes in the SFTS group was lower than those in the IM,HFRS and healthy control groups(P＜0.05).Compared to SFTS,IM and healthy control groups,the lateral scattered light distribution width(NE-WX)and fluorescence distribution width(NE-WY)of neutrophils in the HFRS group were both increased(P＜0.05).CONCLUSIONS There are signifi-cant differences in the characteristics of peripheral blood leukocyte morphology,scatterplot and CPD study param-eters among three viral infection diseases SFTS,IM and HFRS.These differences can provide early clinical advice and assist in clinical differentiation.

Objective:? To summarize the achievements in improving the consistency of clinical liquid chromatography-tandem mass spectrometry (LC-MS/MS) testing results.Methods:? From 2021 to 2024, Zhongshan Hospital Affiliated to Fudan University recruited laboratories voluntarily participating in the MSHP (Clinical LC-MS/MS Testing Consistency Program). As of Batch 202404, a total of 76 laboratories had enrolled, including 60 medical institutions (all tertiary hospitals) and 16 third-party laboratories. Test items were established, and comparative samples were distributed regularly-each item′s samples covered three concentrations (high, medium, and low). Samples were shipped via cold chain and tested within one week. Our laboratory′s measurements served as the target, with participating labs′ results within ±25% of the target deemed qualified. Passing-Bablok regression and Bland-Altman analysis were used to assess consistency.Results:Taking 3-MT (3-methoxytyramine) as an example, the coefficients of variation (CVs) for the project′s three concentration levels improved from 17.00%, 47.18%, and 4.88% in the first comparative batch to 9.59%, 9.59%, and 6.1% in Batch 202404. Passing-Bablok regression results for the 5 units participating in 3-MT testing showed that Laboratory A had proportional bias but no systematic bias (regression slope [95% CI]: 0.903 [0.862-0.952]; intercept [95% CI]: 0.912 [-1.921-6.073]). The remaining laboratories exhibited no proportional or systematic bias with the target (Laboratory B: slope 1.031 [0.961-1.147], intercept-0.733 [-4.641-8.272]; Laboratory C: slope 0.982 [0.940-1.009], intercept-0.576 [-2.675-1.891]; Laboratory D: slope 0.973 [0.939-1.066], intercept-1.168 [-6.108-1.649]; Laboratory E: slope 0.999 [0.905-1.051], intercept-1.876 [-6.111-3.508]). Bland-Altman analysis indicated that all 5 laboratories′ results generally showed good consistency with the target. Through quality feedback and optimizing sample preparation concentrations, result consistency was enhanced.? Conclusion:? Clinical LC-MS/MS testing consistency programs contribute to improving the comparability of test results.

Objective:To assess the prognostic value of methylation of interferon regulatory factor 6 ( IRF6) gene promoter in patients diagnosed with Kidney renal clear cell carcinoma (KIRC). Methods:The primary lesions of fifty KIRC patients who were diagnosed at the First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. The expression of IRF6 protein was determined with an immunohistochemical method. The correlation between the level of IRF6 expression and survival and/or metastasis status was analyzed. The mRNA and protein levels of the IRF6 in KIRC and normal renal tissues were compared by using bioinformatic tools. The difference in the methylation rate of the IRF6 gene promoter between tumor and adjacent tissues was analyzed by searching the online databases. Statistical analysis was carried out for the methylation status of the IRF6 gene promoter region to select those negatively correlated with the overall survival (OS) among the patients. In vitro experiments were conducted with cell lines to verify the correlation between the status of promoter methylation and transcription level of the IRF6 gene. Results:The mRNA and protein levels of the IRF6 gene in KIRC tissues were significantly lower than those of the normal controls, and this was more prominent in patients who had died or developed metastasis. The extent of IRF6 gene promoter methylation in the KIRC tissues was much higher compared with that of the adjacent normal renal tissues. There was a significant negative correlation between the methylation of the IRF6 gene promoter and mRNA level of the IRF6 ( R= -0.52). The higher methylation degree in the IRF6 gene promoter regions cg12034118 and cg16030177, the shorter the OS and worse prognosis in the patients. Only twenty CpG sites in cg12034118 were confirmed to be highly methylated in KIRC cell lines. The transcription level of the IRF6 gene was upregulated in a time- and dose-dependent manner after the treatment with demethylation reagent 5-azadeoxycytidine. Conclusion:The methylation of IRF6 gene promoter in the renal tissues of KIRC patients is closely correlated with the OS. Cg12034118 may provide a promising biomarker for laboratory detection, and its high methylation rate has certain reference value for the prognosis.

Objective To retrieve,evaluate and summarise the relevant evidence in non-pharmacological management of venous thromboembolism in patients with haemorrhagic stroke during perioperative period so as to provide references for clinical practice. Methods According to the "6S" evidence pyramid mode,evidence was retrieved from top to bottom across databases such as BMJ Best Practice,UpToDate,JBI Evidence-Based Healthcare Centre,the National Guideline Clearinghouse (NGC),the National Institute for Health and Care Excellence (NICE),the Scottish Intercollegiate Guidelines Network (SIGN),the Registered Nurses' Association of Ontario (RNAO) guidelines network,the Canadian Heart and Stroke Foundation (HSF),the American Heart Association/American Stroke Association (AHA/ASA),the Neuro-Critical Care Society (NCS),the European Stroke Organization (ESO),Cochrane Library,PubMed,Medline,CNKI,Web of Science,Embase,the China Biomedical Literature Database,Wanfang Data,Medlive,VIP,and the Chinese Medical Journal Full-Text Database. The search focused on non-pharmacological management of perioperative venous thromboembolism in haemorrhagic stroke patients,including guidelines for clinical practice,expert consensus,systematic reviews and evidence summaries. The searched literatrue was from the inception of the databases to April 11th,2024. Two researchers independently evaluated the quality of the literature,extracted,integrated and summarised the best evidences. Results A total of 23 articles were included,comprising 12 guidelines,5 systematic reviews,4 expert consensuses and 2 evidence summaries. Thirty-one pieces of the best evidence were summarised and integrated into 5 topics,including multidisciplinary teamwork,risk assessment and screening,basic prevention,mechanical prophylaxis and health education. Conclusion This study has summarised the best evidence for non-pharmacological management of venous thromboembolism in patients with haemorrhagic stroke during the perioperative period. It provides evidence-based references for clinical medical staff to prevent and manage venous thromboembolism in patients with hemorrhagic stroke during perioperative period.

Objective To retrieve,evaluate and summarise the relevant evidence in non-pharmacological management of venous thromboembolism in patients with haemorrhagic stroke during perioperative period so as to provide references for clinical practice. Methods According to the "6S" evidence pyramid mode,evidence was retrieved from top to bottom across databases such as BMJ Best Practice,UpToDate,JBI Evidence-Based Healthcare Centre,the National Guideline Clearinghouse (NGC),the National Institute for Health and Care Excellence (NICE),the Scottish Intercollegiate Guidelines Network (SIGN),the Registered Nurses' Association of Ontario (RNAO) guidelines network,the Canadian Heart and Stroke Foundation (HSF),the American Heart Association/American Stroke Association (AHA/ASA),the Neuro-Critical Care Society (NCS),the European Stroke Organization (ESO),Cochrane Library,PubMed,Medline,CNKI,Web of Science,Embase,the China Biomedical Literature Database,Wanfang Data,Medlive,VIP,and the Chinese Medical Journal Full-Text Database. The search focused on non-pharmacological management of perioperative venous thromboembolism in haemorrhagic stroke patients,including guidelines for clinical practice,expert consensus,systematic reviews and evidence summaries. The searched literatrue was from the inception of the databases to April 11th,2024. Two researchers independently evaluated the quality of the literature,extracted,integrated and summarised the best evidences. Results A total of 23 articles were included,comprising 12 guidelines,5 systematic reviews,4 expert consensuses and 2 evidence summaries. Thirty-one pieces of the best evidence were summarised and integrated into 5 topics,including multidisciplinary teamwork,risk assessment and screening,basic prevention,mechanical prophylaxis and health education. Conclusion This study has summarised the best evidence for non-pharmacological management of venous thromboembolism in patients with haemorrhagic stroke during the perioperative period. It provides evidence-based references for clinical medical staff to prevent and manage venous thromboembolism in patients with hemorrhagic stroke during perioperative period.

Pancreatic cancer is a disease with a high fatality rate， with a five-year survival rate of no more than 10% and a significantly increasing annual mortality rate. The common pathogenesis factors of pancreatic cancer are family inheritance， diet， pancreatitis， obesity， etc.， among which， family inheritance of pancreatic cancer is the main reason， and about 7%-10% of patients have family inheritance. Surgery is an effective way to treat pancreatic cancer in patients and improve their survival， but most people are diagnosed with pancreatic cancer at intermediate and advanced stages and lose the opportunity for surgical treatment. Therefore， radiotherapy， interventional therapy， supportive treatment， immunotherapy， and other treatments are used clinically to relieve symptoms and prolong the survival of patients. The commonly used clinical drug is gemcitabine. Although it can inhibit tumor growth and improve the condition， it can bring side effects such as bone marrow suppression， rash， digestive tract side effects， and drug resistance. The damage of these side effects to the human body is systemic. Chinese medicine can be used alone or in combination with other treatment methods to reduce the toxic and side effects of chemotherapy， restore the physical energy of patients， and reduce its related complications. Chinese medicine contains a large number of active ingredients， including alkaloids， flavonoids， saponins， phenolic acids， and organic acids， with anti-inflammatory， anti-viral， anti-tumor， and other curative effects. Many clinical studies of traditional Chinese medicine （TCM） on cancers have verified that TCM plays a positive role in tumor prevention and treatment， especially in improving and controlling clinical symptoms， and also plays a good detoxification effect on radiotherapy and chemotherapy， with good results achieved in improving bone marrow suppression， improving immunity， improving quality of life， and prolonging survival. This paper reviewed the anti-pancreatic cancer mechanism of Chinese medicine monomers based on literature in China and abroad， aiming to provide new potential drug candidates for the treatment of pancreatic cancer.

ObjectiveTo explore the meridian tropism of components in Bupleuri Radix （Chaihu， CH） based on the model of nonalcoholic steatohepatitis （NASH） and clarify the substance basis of the meridian tropism of CH in Xiaoyaosan （XYS） by means of principal component analysis. MethodEighty SPF male C57BL/6 mice were randomly assigned into 8 groups， with 10 mice in each group. Except that the blank group was fed with the methionine choline-sufficient （MCS） diet， the other mice were fed with methionine choline-deficient （MCD） diet for 4 weeks to establish the nonalcoholic steatohepatitis （NASH） model. After the established model was confirmed by hematoxylin-eosin （HE） staining， the mice were administrated with corresponding drugs by gavage once a day for 4 weeks. Specifically， the 8 groups were XYS group （2.874 g·kg^-1）， XYS-CH group （2.445 g·kg^-1）， XYS-CH+volatile oils （Vol， 0.163 mg·kg^-1） group， XYS-CH+polysaccharides （Pol， 24.067 mg·kg^-1） group， XYS-CH+flavones （Fla， 2.241 mg·kg^-1） group， and XYS-CH+saponins （Sap， 2.746 mg·kg^-1） group. The model group and the blank group were administrated with the same volume of normal saline. After the last administration， the mice were sacrificed for the collection of blood and liver tissue. The pathological changes of liver were observed by HE staining and oil red O staining. Enzyme linked immunosorbent assay （ELISA） kits were used to determine the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein （HDL）， and low-density lipoprotein （LDL） in serum as well as malondialdehyde （MDA）， superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） in liver. SPSS Statistics 23 was used for principal component analysis and comprehensive evaluation to determine the substance basis of the meridian tropism of CH in NASH mice. ResultCompared with the blank control group， the modeling led to hepatocyte swelling， increased fat vacuoles， and appearance of inflammatory cells. Further， the modeling elevated the levels of ALT， AST， TG， TC， and LDL and lowered the HDL level in serum， and it increased the MDA level and decreased the SOD， CAT， and GSH-Px levels in liver. Compared with the model group， the administration of XYS and XYS-CH in combination with the components of CH alleviated the oxidative damage in liver （P<0.05）. The comprehensive score of the pharmacological efficacy was in a descending order as follows： XYS > XYS-CH+Sap > XYS-CH+Fla > XYS-CH+Pol > XYS-CH+Vol > XYS-CH. Among the chemical components of CH， Sap had the best effect. ConclusionSap lowers the blood lipid level， regulates the abnormal lipid metabolism， and alleviates the oxidative damage of liver， which is the substance basis for CH to exert the meridian tropism in liver.