ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

ObjectiveTo investigate the effect and mechanism of transplantation of human umbilical cord mesenchymal stem cell （hUC-MSC） with overexpression of the Numb gene in the treatment of cholestatic liver fibrosis （CLF）. MethodsThe technique of lentiviral transfection was used to induce the overexpression of the Numb gene in hUC-MSC （hUC-MSC^Numb-OE）， and hUC-MSC transfected with empty vector （hUC-MSC^OE-EV） was used as negative control. Bile duct ligation （BDL） was performed to establish a rat model of CLF， and then the rats were randomly divided into BDL group， hUC-MSC group， hUC-MSC^OE-EV group， and hUC-MSC^Numb-OE group， while a sham-operation group was also established. The rats in the intervention groups were given a single splenic injection of the corresponding cells after BDL， and samples were collected at the end of week 4. Related indicators were measured， including serum biochemistry， liver histopathology， the content of hydroxyproline （Hyp） in the liver， hepatic stellate cell activation， ductular reaction， liver regeneration， and the expression levels of key molecules in the Numb-p53 signaling axis. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the BDL group， the hUC-MSC group and the hUC-MSC^OE-EV group had significant reductions in the levels of serum biochemical parameters （aspartate aminotransferase， gamma-glutamyl transpeptidase， total bile acid， total bilirubin， and direct bilirubin）， liver fibrosis markers （the content of Hyp and the expression levels of alpha-smooth muscle actin， tumor necrosis factor-α， and transforming growth factor-beta 1）， and ductular reaction markers （the expression levels of CK7 and CK19） （all P <0.05）， and compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significantly greater improvements in the above indicators （all P <0.05）. In addition， compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significant improvements in the expression levels of liver regeneration-related markers （albumin and hepatocyte nuclear factor 4α） and the molecules associated with the Numb-p53 signaling axis （Numb， pNumb， Mdm2， and p53） （all P <0.05）. ConclusionOverexpression of the Numb gene can enhance the therapeutic effect of hUC-MSC on CLF， possibly by activating the Numb-PTB^L-p53-HNF4α axis， promoting the hepatic differentiation of hUC-MSCs and subsequently enhancing liver regeneration.

ObjectiveTo investigate the epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer by regulating DNA methylation， and to provide a basis for scientific clinical medication. MethodsA total of 48 specific pathogen-free Sprague-Dawley rats were divided into normal group， model group， and Diwu Yanggan Capsule group using a random number table， with 16 rats in each group. The Solt-Farber two-step method was used to establish a rat model of liver cancer. The rats in the Diwu Yanggan Capsule group were given Diwu Yanggan Capsule at a dose of 750 mg/kg/d by gavage， and those in the normal group and the model group were given an equal volume of normal saline by gavage. Liver tissue samples were collected from each group of rats after 16 weeks of continuous intervention； DNA methylation chips were used to analyze the change in DNA methylation in liver tissue， and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were used for data analysis. In addition， the MeDIP-PCR technique was used to detect the changes in candidate differentially methylated genes such as YWHAB， ADCK2， ERLIN2， SEMA3B， and TPH2 in the liver tissue of rats， and Western blot and RT-qPCR were used to verify the expression of key methylated genes. The independent-samples t test was used for comparison of continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups， while the least significant difference t-test was used for further comparison between two groups. ResultsThe DNA methylation chip analysis showed that compared with the normal group， the model group had significant methylation changes in the promoter region of 2 422 genes in liver tissue of rats. The GO functional enrichment analysis and the KEGG pathway enrichment analysis showed that these differentially methylated genes were significantly enriched in metabolic pathways such as steroid hormone biosynthesis and drug metabolism-cytochrome P450. Compared with the model group， the Diwu Yanggan Capsule group had significant reversal of promoter methylation in 1 650 genes， and the KEGG enrichment analysis showed that these genes were mainly involved in the pathways closely associated with cell proliferation， apoptosis， and microenvironment regulation， such as the calcium ion signaling pathway， the cAMP signaling pathway， and the extracellular factor signaling pathway. Compared with the model group， the Diwu Yanggan Capsule group had a significant increase in the promoter methylation level of the ADCK2 gene （P<0.05） and significant reductions in the promoter methylation levels of the ERLIN2 and TPH2 genes （all P<0.05）. Compared with the model group， the Diwu Yanggan Capsule group had significant reductions in the mRNA expression levels and the protein expression levels of the ADCK2 （all P<0.05）. ConclusionAbnormal DNA methylation in liver tissue participates in the development and progression of liver cancer. The effect of Diwu Yanggan Capsule on DNA methylation level is an important epigenetic mechanism for its effect in the prevention and treatment of liver cancer.

Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

A 53-year-old female patient with type 2 diabetes mellitus,hypertension,and hyperlipidemia,on long-term therapy with metformin,gliclazide,dapagliflozin,sacubitril/valsartan,lercanidipine,and atorvastatin,was initiated on semaglutide due to obesity and suboptimal glycemic control.Patient using semaglutide 0.25 mg and 0.5 mg without adverse effects,and developed gastric discomfort after adjusting dose to 0.75 mg.She remained on this dose for 7 weeks.Upon further dose escalation to 1 mg,she experienced persistent nausea,vomiting,and diarrhea.Fifteen days later,she was admitted to the hospital with impaired consciousness.Laboratory findings revealed blood urea nitrogen of 35.4 mmol·L-1 and serum creatinine of 825 μmol·L-1.The patient was diagnosed with acute kidney injury(AKI),which was considered related to semaglutide,metformin,dapagliflozin,and sacubitril/valsartan.All medications were discontinued.Following symptomatic treatment including fluid resuscitation,volume expansion,and hemodialysis,the patient's renal function gradually recovered.The association between AKI and semaglutide,metformin,dapagliflozin,and sacubitril/valsartan was evaluated using the Naranjo's Assesment Scale,the results were all"probable".This case highlighted that clinical use of semaglutide requires careful consideration of concomitant medications and vigilance for renal impairment.In the event of AKI,prompt assessment,discontinuation of medications with potential renal impairment and symptomatic management were necessary to ensure safe medication administration.

Objective To investigate the effect of body position angle on gastric insufflation during the induction of general anesthesia in pediatric patients undergoing day surgery.Methods A total of 111 children scheduled for elective tracheal intubation under general anesthesia at the First Affiliated Hospital of Guangxi Medical University from December 2022 to March 2023 were selected as study subjects.Six children were ex-cluded due to unclear sonographic visualization of the gastric antrum,resulting in 105 children ultimately in-cluded in the study.The children were divided into three groups using a random number table method:Group D0(supine position),Group D5(5° head-up position),and Group D10(10° head-up position),with 35 children in each group.The presence of a"comet-tail artifact"on ultrasound was used as the criterion for determining gastric insufflation during anesthesia induction.The gastric antrum cross-sectional area(CSA)of the gastric antrum was measured before mask ventilation after loss of consciousness(T1)and immediately after tracheal intubation following mask ventilation(T2).The incidence of gastric insufflation,changes in CSA and their differences,and vital sign changes at T1 and T2,were compared among the three groups.Results Compared with Group D0,the incidence of gastric insufflation was significantly lower in Groups D5 and D10[25.7％(9/35)vs.20.0％(7/35)vs.54.0％(19/35)],and the difference was statistically significant(P＜0.05).Howev-er,there was no significant difference between Group D5 and Group D10(P＞0.05).No significant difference was observed in CSA at T1 among the three groups(P＞0.05).At T2,a statistically significant difference in CSA was found among the three groups(P＜0.05),with Group D10 showing a smaller CSA than Group D0(P＜0.05).The difference in CSA changes was statistically significant among the three groups(P＜0.05),with Groups D5 and D10 exhibiting smaller changes than Group D0,and Group D10 showing a smaller change than Group D5(P＜0.05).No significant differences were observed in mean artery pressure(MAP),heart rate,pulse oxygen saturation(SpO2)and pressure of end-tidal carbon dioxide(PETCO2)at T1 and T2 among the three groups(P＞0.05).A total of 6 children(5.7％)required brief adjustments in jaw support due to irregular PETCO2 waveforms or abnormal peak airway pressure.Conclusion In pediatric patients undergoing day sur-gery under general anesthesia,a 5° head-up position during induction significantly reduces the incidence of gas-tric insufflation and minimizes changes in gastric antral CSA,while a 10° head-up position does not provide ad-ditional benefits.

OBJECTIVE To analyze the value of elastic modulus of shear wave elastography in the diagnosis of cervical lymph node metastasis of thyroid cancer and its correlation with VEGF-C and VEGF-D.METHODS A total of 120 patients with thyroid cancer admitted to our hospital were divided into metastatic group(n=37)and non-metastatic group(n=83)according to whether the cervical lymph node metastasis.The clinical data and SWE elastic modulus values of the two groups were compared,and the effects of clinical characteristics on SWE elastic modulus values were analyzed by hierarchical regression.Logistic regression analysis was conducted to analyze the independent correlation between SWE elastic modulus and the risk of lymph node metastasis of thyroid cancer.The dose-response relationship between the elastic modulus of SWE and the risk of lymph node metastasis in thyroid cancer was analyzed with restricted cubic strips.The correlation between SWE elastic modulus and VEGF-C and D levels was analyzed after adjusting factors by Logistic regression.RESULTS The results of general data analysis showed that male proportion,BMI,clinical stage,lesion diameter,TSH,FT4,TgAb positive rate,TPOAb positive rate,VEGF-C and VEGF-D in metastatic group were higher than those in non-metastatic group.Under different clinical characteristics,SWE parameters Emax,Emean and Eratio in metastatic group were higher than those in non-metastatic group.In hierarchical regression analysis,gender,clinical stage,lesion diameter,biochemical indices(TSH,FT4,TgAb,TPOAb)had significant positive effects on SWE parameters.SWE parameters were independently associated with the risk of lymph node metastasis of thyroid cancer.Subgroup analysis showed that the association between SWE parameters and the risk of lymph node metastasis of thyroid cancer in all subgroups was statistically significant(P<0.05).There was no nonlinear dose-response relationship between SWE parameters and thyroid cancer risk(P>0.05).After adjusting for confounding factors,there was a significant linear correlation between Emax,Emean,Eratio and VEGF-C and VEGF-D(P<0.05).CONCLUSION The elastic modulus of SWE has a certain diagnostic value for lymph node metastasis of thyroid cancer,and SWE parameters have a certain correlation with VEGF-C and VEGF-D.

Risk prediction models for postoperative pulmonary complications (PPCs) can assist healthcare professionals in assessing the likelihood of PPCs occurring after surgery, thereby supporting rapid decision-making. This study evaluated the merits, limitations, and challenges of these models, focusing on model types, construction methods, performance, and clinical applications. The findings indicate that current risk prediction models for PPCs following lung cancer surgery demonstrate a certain level of predictive effectiveness. However, there are notable deficiencies in study design, clinical implementation, and reporting transparency. Future research should prioritize large-scale, prospective, multi-center studies that utilize multiomics approaches to ensure robust data for accurate predictions, ultimately facilitating clinical translation, adoption, and promotion.

Objective The study aimed to explore the mechanism of Mume Fructus and Rosa multiflora in the treatment of recurrent oral ulceration(ROU)through network pharmacology and animal experiments.Methods The chemical components of Mume Fructus and Rosa multiflora were filtrated by OB and DL,which were respectively searched from TCMSP and references.The targets were predicted by TCMSP and Swiss Target Prediction.The targets of ROU were searched from OMIM and GeneCards database.Cystoscape 3.2.1 software was used to construct the"components-targets"network and analysis gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway with Clue GO.The protein-protein interaction(PPI)network analysis was carried out on the common targets by String database and Cystoscape 3.2.1 software.Moreover,after constructing the rat model of ROU,the pathological changes of oral mucosa of rats were observed by HE staining,and the effects of Mume Fructus and Rosa multiflora on inflammatory factors and key targets were detected by ELISA and Western blot.Results 33 active ingredients were selected for treating ROU,among which quercetin,ursolic acid,β-sitosterol,stigmasterol,palmitic acid,apioline and kaempferol were the core components.Key targets such as STAT3,PIK3R1,PIK3CA,STAT1,JAK2 and IL-6 were screened.GO functional enrichment and KEGG enrichment analysis showed that the core targets mainly affected the process of response to organic substance,cellular response to chemical stimulus and cellular response to organic substance,involving NF-κB,JAK-STAT,IL-17 and other signaling pathways.Experimental results in rats showed that Mume Fructus and Rosa multiflora could significantly improve the pathological status of ulcer mucosal tissue,reduce the expression of IL-1β,IL-6,TNF-α and IFN-γ,and reduce the expression of JAK2 and STAT1 in oral mucosa.Conclusion The anti-ROU effect of Mume Fructus and Rosa multiflora may be achieved by regulating the JAK2-STAT1 signaling pathway,reducing the levels of IL-1β,IL-6,TNF-α and IFN-γ inflammatory factors.

A 53-year-old female patient with type 2 diabetes mellitus,hypertension,and hyperlipidemia,on long-term therapy with metformin,gliclazide,dapagliflozin,sacubitril/valsartan,lercanidipine,and atorvastatin,was initiated on semaglutide due to obesity and suboptimal glycemic control.Patient using semaglutide 0.25 mg and 0.5 mg without adverse effects,and developed gastric discomfort after adjusting dose to 0.75 mg.She remained on this dose for 7 weeks.Upon further dose escalation to 1 mg,she experienced persistent nausea,vomiting,and diarrhea.Fifteen days later,she was admitted to the hospital with impaired consciousness.Laboratory findings revealed blood urea nitrogen of 35.4 mmol·L-1 and serum creatinine of 825 μmol·L-1.The patient was diagnosed with acute kidney injury(AKI),which was considered related to semaglutide,metformin,dapagliflozin,and sacubitril/valsartan.All medications were discontinued.Following symptomatic treatment including fluid resuscitation,volume expansion,and hemodialysis,the patient's renal function gradually recovered.The association between AKI and semaglutide,metformin,dapagliflozin,and sacubitril/valsartan was evaluated using the Naranjo's Assesment Scale,the results were all"probable".This case highlighted that clinical use of semaglutide requires careful consideration of concomitant medications and vigilance for renal impairment.In the event of AKI,prompt assessment,discontinuation of medications with potential renal impairment and symptomatic management were necessary to ensure safe medication administration.