Liver regeneration following injury aids the restoration of liver mass and the recovery of liver function. In the present study we investigated the contribution of megakaryocytic leukemia 1 (MKL1), a transcriptional modulator, to liver regeneration. We report that both MKL1 expression and its nuclear translocation correlated with hepatocyte proliferation in cell and animal models of liver regeneration and in liver failure patients. Mice with MKL1 deletion exhibited defective regenerative response in the liver. Transcriptomic analysis revealed that MKL1 interacted with E2F1 to program pro-regenerative transcription. MAPKAPK2 mediated phosphorylation primed MKL1 for its interaction with E2F1. Of interest, phospholipase d2 promoted MKL1 nuclear accumulation and liver regeneration by catalyzing production of phosphatidic acid (PA). PA administration stimulated hepatocyte proliferation and enhanced survival in a MKL1-dependent manner in a pre-clinical model of liver failure. Finally, PA levels was detected to be positively correlated with expression of pro-regenerative genes and inversely correlated with liver injury in liver failure patients. In conclusion, our data reveal a novel mechanism whereby MKL1 contributes to liver regeneration. Screening for small-molecule compounds boosting MKL1 activity may be considered as a reasonable approach to treat acute liver failure.

ObjectiveTo determine the pharmacodynamic substance basis of Epimedii Folium（EF） and Epimedii Wushanensis Folium（EWF） in promoting osteogenic differentiation， and to establish a method to analyze the material basis of Chinese materia medica based on the correlation between chemical fingerprint and cellular metabolomics. MethodThe chemical fingerprints of 15 batches of EF with 4 species and 3 batches of EWF were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， and partial least squares-discriminant analysis（PLS-DA） was used to analyze the peak areas of chemical fingerprints of samples. The effects of different samples on proliferative activity of MC3T3-E1 osteoblast precursors， as well as the activity of alkaline phosphatase（ALP） in osteoblasts were detected by cell counting kit-8（CCK-8） and enzyme-linked immunosorbent assay（ELISA）. At the same time， UPLC-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was used to analyze the effects of different samples on the metabolomics of MC3T3-E1 cells， then metabolic peak table of osteogenic differentiation cells was constructed， and pharmacodynamic index mean Y₀ was introduced into the peak table. PLS was used to calculate mean Y₀ of each group， and the mean Y₀ was added to the peak table of chemical fingerprint to construct the correlation between chemical fingerprint and cell metabolome， the pharmacodynamic components of EF and EWF that promote bone differentiation were screened according to variable importance in the projection（VIP） value>1. The pharmacodynamic effects of EF and EWF were evaluated according to the mean Y₀ of each group. ResultThe chemical fingerprints of EF with different origins and EWF were completely separated. Compared with the blank group， the activity of MC3T3-E1 cells in EF and EWF groups was significantly increased， the activity of ALP in the Epimedium brevicornu（Gansu province）， E. koreanum and E. pubescens groups was significantly increased（P<0.05）. The results of cell metabolomics showed that the blank group and the model group had an obvious trend of separation. EF with different origins and EWF had different distance from the model group， indicating that EF with different origins and EWF had different effect on promoting osteogenic differentiation. Chemical fingerprint-cell metabolomics integration analysis screened 9 components closely related to the efficacy of EF and EWF， including diphylloside B， epimedin C， icariin， baohuoside Ⅰ， yinyanghuo B， β-anhydroicaritin， magnoflorine， cryptochlorogenic acid and quercetin. E. koreanum had the strongest effect on promoting osteogenic differentiation. ConclusionThis study determined that the material basis of EF and EWF promoting osteogenic differentiation were mostly flavonoids， alkaloids and organic acids， which provided ideas and methods for the screening of pharmacodynamic components and the prediction of therapeutic effect of Chinese materia medica.

Objective:To analyze the risk factors of multidrug-resistant organism（MDRO）infection among patients in neurosurgery intensive care unit（NSICU）and to construct a risk prediction nomogram.Methods:A total of 434 patients admitted in the NSICU of the First Affiliated Hospital of Kangda College of Nanjing Medical University from August 2021 to October 2022 were enrolled in the study. Patients were divided into modeling group（ n=217）and validation group（ n=217）. Multivariate Logistic regression was used to analyze the risk factors of MDRO infection in patients，and R software was used to construct a risk prediction nomogram.The prediction power of the nomogram was evaluated with receiver operating characteristic（ROC）curve，the calibration of the model was assessed with Hosmer-Lemeshow（H-L）goodness-of-fit method. Results:Multivariate Logistic regression analysis showed that the underlying disease≥3（ OR=2.580，95% CI 1.322-5.035），the combination of antimicrobial drugs >10 days（ OR=2.336，95% CI 1.182-4.615），hypoproteinemia（ OR=1.962，95% CI 1.031-3.735），invasive operation time（10-20 d： OR=2.358，95% CI 1.048-5.306；>20 d： OR=3.486，95% CI 1.643-7.395）and GCS≤8 points（ OR=2.961，95% CI 1.470-5.963）were independent risk factors of MDRO infection among patients in NSICU. The area under the curve（AUC）of the nomogram in predicting the risk of MDRO infection for patients in modeling group was 0.787（95% CI 0.725-0.849）with a sensitivity of 73.3% and a specificity of 72.5%，the H-L test results were χ2=7.482， P=0.486，the calibration curve was close to the ideal curve，and the mean absolute error was 0.022. The AUC of the nomogram in predicting MDRO infection for patients in verification group was 0.800（95% CI 0.739-0.861）with a sensitivity of 74.7% and a specificity of 73.9%，the H-L test results were χ2=9.824， P=0.278，the calibration curve was close to the ideal curve，and the average absolute error was 0.015. Conclusion:The nomogram constructed based on the risk factors can effectively predict the risk of MDRO infection for patients in neurosurgical ICU，which may be used in clinic pratcice.

Objective To summarize the clinical characteristics of children with drug-induced hypersensitivity syndrome(DIHS)complicated with herpesvirus reactivation,and to promote the early and accurate identification,diagnosis,and treatment of DIHS in children.Methods The medication history,clinical manifestations,treatment,and prognosis of 12 children confirmed DIHS complicated with herpesvirus reactivation in Guangzhou Women and Children's Medical Center between January 2018 and March 2023 were retrospectively analyzed.The changes in hematological parameters,inflammatory indexes,and hepatic and renal function within 5 d before the eruption,5 d,and 6-10 d after eruption were compared.Results Of the 12 patients,the male-to-female ratio was 5∶1,with a median age of 27(interquartile range 20.50-34.75)months.Two or more antibiotics were used at least two to six weeks before onset,with a combination of 3 or more antibiotics in 7 children,and a combined or sequential application of 2 antibiotics in 5 children.The antibiotics included cephalosporins(n=12),semisynthetic penicillins(n=5),vancomycin(n=4)and azithromycins(n=7).All 12 patients presented fever,rashes,and multiple organ involvement.The rashes were red maculopapules in the early stage and then gradually developed into massive fusion exceeding 50%of the whole body.Among them,seven children were accompanied by facial edema,and two had purplish-red facial rashes.11 children suffered from exfoliative dermatitis in the later stage.12 children presented obviously enlarged lymph nodes.Liver involvement was the most common(100%,simple increase of transaminase in four children,cholestasis in six children,and hepatic failure in two children),and lung involvement was found in nine children.Laboratory examination showed no significant increase in leukocytes or eosinophils within 5 d before the eruption,but low levels of atypical lymphocytes.After the eruption,leukocytes,eosinophils,and atypical lymphocytes increased progressively.Inflammatory indexes of hypersensitive C-reactive protein(CRP),procalcitonin(PCT)increased dramatically before and after the eruption.All the children received intravenous immunoglobulin(IVIG)and methylprednisolone,two children were given antiviral therapy,and nine children were treated with multiple plasma exchanges.After treatment,nine children were cured,one developed immune reconstitution syndrome,and two died of hepatic failure.Conclusions Antibiotics are common allergenic drugs for DIHS in children.Its clinical manifestations include fever and rashes,accompanied by multiple organ involvement,such as the liver and lung.When leukocytes,eosinophils,and atypical lymphocytes are progressively elevated after the eruption,DIHS should be highly suspected,herpesvirus activation should be monitored,medication history should be traced,and early active immunotherapy and antiviral therapy should be conducted if necessary.

Objective To investigate the effect of melanoma associated antigen D1 (Mage-D1)on mouse femoral bone mass and mineralization ability of mouse bone marrow mesenchymal cells (BMSCs)and its potential molecular mechanism.Methods Female Mage-D1 gene knockout heterozygous mice and male wild-type (WT)mice were subjected as parent mice to breed Mage-D1 gene knockout homozygous (Mage-D1 KO)mice.PCR and agarose gel electrophoresis were used to identify male Mage-D1 knockout (Mage-D1 KO)mice and littermate male wild-type (WT)mice.Micro-CT scanning was performed to observe mouse femoral bone mass,and ELISA and chemical assay were employed to detect serum levels of calcium,phosphorus,calcitonin,and parathyroid hormone in mice.After primary cultured BMSCs were identified with flow cytometry,immunofluorescence staining was utilized to detect the expression of Mage-D1 in BMSCs.BMSCs were infected by Mage-D1 silencing lentivirus,and then the cells were divided into negative control group (sh-NC)and silencing group (sh-Mage-D1).Cell scratch assay was conducted to detect the migration ability of BMSCs,and flow cytometry and CCK-8 assay were conducted to detect the cycle change and proliferation ability of BMSCs.After mineralization induction,alkaline phosphatase (ALP) staining and alizarin red staining were performed;RT-qPCR and Western blotting were used to measure the expression levels of ALP,Runx2 and Col1.RT-qPCR was used to detect mineralization-related genes p75NTR and Msx1.Results Compared with the WT mice,the femoral cortical bone thickness,cortical bone mineral content,cancellous bone mineral content,trabecular number,and cancellous bone surface density were decreased,and trabecular separation was increased in the Mage-D1 knockout homozygous mice (P<0.05).There were no significant changes in the serum levels of calcium,phosphorus,calcitonin and parathyroid hormone in mice after Mage-D1 knockout.Mage-D1 was expressed in the whole BMSCs and was highly expressed in the nucleus and perinuclear regions.Compared with the sh-NC BMSCs,the sh-Mage-D1 group had decreased proliferation ability (P<0.01),enhanced migration ability (P<0.01),and decreased expression of ALP,Runx2 and Col1 genes (P<0.05)and protein (P<0.01)after mineralization induction,milder ALP and alizarin red stain,and lower expression levels of p75NTR and Msx1.Conclusion Mage-D1 knockout can significantly reduce femur bone mass in mice.It can promote the proliferation and inhibit migration of BMSCs,and positively regulate their mineralization in vitro,and the p75NTR-Dlx1/Msx1 signaling axis may be involved in the regulation of bone metabolism by Mage-D1.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

OBJECTIVE To compare efficacy and safety of continuous pump versus intermittent infusion of amphotericin B in the treatment of invasive fungal infection， and to provide evidence-based reference for clinical treatment. METHODS Retrieved from PubMed， the Cochrane Library， Web of Science， Embase， Wanfang， CNKI， CBM and VIP database， randomized controlled trial （RCT） and cohort study about 24 h continuous pump （trial group） versus intermittent infusion （control group） of amphotericin B were collected from the inception to Jan. 2023. After literature screening and data extraction， the quality of RCT was evaluated with modified Jadad scale， and the quality of cohort study was evaluated with Newcastle-Ottawa scale. Meta-analysis and sensitivity analysis were performed by using RevMan 5.4 software. RESULTS A total of 7 literature were included， involving 1 RCT and 6 cohort studies with a total of 767 patients. The results of meta-analysis showed that the clinical effective rate [RR＝1.44， 95%CI （1.13，1.83）， P＝0.003] of trial group was significantly higher than that of control group， and all-cause mortality rate [RR＝0.37， 95%CI（0.19，0.72），P＝0.003] and the incidence rate of infusion reaction [RR＝0.28，95%CI（0.18，0.43）， P＜0.000 01] were significantly lower than control group； there was no statistical significance in the incidence rate of renal impairment between 2 groups [RR＝0.71，95%CI（0.45，1.11），P＝0.13] . The sensitivity analysis results showed that the results obtained in this study were robust. CONCLUSIONS The efficacy and safety of 24 h continuous pump of amphotericin B are better than those of intermittent infusion in the treatment of invasive fungal infection.

Sepsis is a complex syndrome caused by multiple pathogens and involves multiple organ failure, particularly spleen dysfunction. In 2017, the worldwide incidence was 48.9 million sepsis cases and 11 million sepsis-related deaths were reported (Rudd et al., 2020). Inflammation, oxidative stress, and apoptosis are the most common pathologies seen in sepsis. Liensinine (LIE) is a bisbenzylisoquinoline-type alkaloid extracted from the seed embryo of Nelumbo nucifera. Lotus seed hearts have high content of LIE which mainly has antihypertensive and antiarrhythmic pharmacological effects. It can exert anti-carcinogenic activity by regulating cell, inflammation, and apoptosis signaling pathways (Manogaran et al., 2019). However, its protective effect from sepsis-induced spleen damage is unknown. In this research, we established a mouse sepsis model induced by lipopolysaccharide (LPS) and investigated the protective effects of LIE on sepsis spleen injury in terms of inflammatory response, oxidative stress, and apoptosis.
Mice ; Animals ; Lipopolysaccharides/pharmacology* ; Spleen ; Inflammation ; Apoptosis ; Sepsis ; Oxidative Stress

Mycolic acids (MAs), i.e. 2-alkyl, 3-hydroxy long-chain fatty acids, are the hallmark of the cell envelope of Mycobacterium tuberculosis and are related with antibiotic resistance and host immune escape. Nowadays, they've become hot target of new anti-tuberculosis drugs. There are two main methods to detect MAs, ¹⁴C metabolic labeling thin-layer chromatography (TLC) and liquid chromatograph mass spectrometer (LC-MS). However, the user qualification of ¹⁴C or the lack of standards for LC-MS hampered the easy use of this method. TLC is a common way to analyze chemical substance and can be used to analyze MAs. In this study, we used tetrabutylammonium hydroxide and methyl iodide to hydrolyze and formylate MAs from mycobacterium cell wall. Subsequently, we used diethyl ether to extract methyl mycolate. By this method, we can easily extract and analyze MA in regular biological labs. The results demonstrated that this method could be used to compare MAs of different mycobacterium in different growth phases, MAs of mycobacteria treated by anti-tuberculosis drugs or MAs of mycobacterium mutants. Therefore, we can use this method as an initial validation for the changes of MAs in researches such as new drug screening without using radioisotope or when the standards are not available.
Mycolic Acids/metabolism* ; Chromatography, Thin Layer ; Mycobacterium tuberculosis ; Fatty Acids ; Antitubercular Agents/pharmacology*

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.