Objective: To evaluate the efficacy and safety of plasma exchange (PE) in thymoma-associated myasthenia gravis (MG), thereby to provide theoretical support for its application in the treatment of thymoma-associated MG. Methods: A total of 133 patients with thymoma-associated MG admitted from January 2018 to September 2024 were retrospectively analyzed. Patients were matched using propensity score to reduce selection bias, yielding 22 matched pairs for both PE group (n=22) and non-PE group (n=22). Patient characteristics including gender, age of disease onset, course of disease, history of thymoma resection, clinical absolute scores [clinical absolute scores (CAS) and clinical relative scores (CRS)], and synchronized immunotherapy regimen of the two groups were analyzed. The CAS scores before and after treatment were compared between the two groups, and the CRS was used to assess the treatment efficiency. Safety of the two treatment regimens were also compared. Continuous variables were compared using the t-test or ANOVA, while categorical data were compared by the chi-square test. Results: A total of 133 patients were included and divided into two groups according to whether they underwent plasma exchange treatment: the PE group (n=22) and the non-PE group (n=111). To exclude bias caused by large difference in the number of cases between the two groups, we performed propensity score matching. After matching, the number of cases in both groups was 22. There was no significant difference in baseline clinical characteristics between the two groups (P>0.05), including gender, age of onset, duration of disease course, history of thymectomy and baseline CAS score before treatment. Compared to the non-PE group, patients in the PE group showed more significant improvement in CAS score (5.09±1.95 vs 3.59±1.50, P<0.05) and a higher CRS score (75.00% vs 50.00%, P<0.001). Compared to the non-PE group, PE group had significantly longer ICU stay, longer hospital stay and higher hospitalization cost (P<0.05). There was no statistically significant difference in adverse events between the two groups during treatment (P>0.05). During long-term follow-up, both the PE and non-PE groups showed relatively low 1-, 3-, and 5-year recurrence rate, with no significant difference between the two groups (P>0.05). Conclusion: This study indicates that plasma exchange has clear value in the treatment of patients with thymoma-associated myasthenia gravis. It can not only significantly improve patients' muscle strength to alleviate motor dysfunction and enhance quality of life, but also does not significantly increase the incidence of adverse reactions. Therefore, it can be regarded as one of the preferred treatment options that achieve a "balance between efficacy and safety" for such patients, and provides an important basis for optimizing treatment strategies, improving prognosis, and promoting the application of subsequent treatment regimens.

A serum metabolomics analysis method based on gas chromatography-mass spectrometry(GC-MS) was used to investigate the metabolic regulation mechanism of Hericium erinaceus(H. erinaceus) polysaccharides on radiation injury. A mouse model of radiation injury was established by ~(60)Co-γ irradiation. High and low dose groups of H. erinaceus polysaccharide injection were designed, and Rubiae Radix et Rhizoma extract was set as the positive control group to investigate the therapeutic effects and metabolic reaction pathways of H. erinaceus polysaccharides on radiation injury. The metabolites of serum samples were collected by GC-MS, and principal component analysis(PCA) was conducted to establish the metabolic profiles of each group of mice. Partial least squares discriminant analysis(PLS-DA), t-test(P<0.05), and variable importance in the projection(VIP>1) were used to screen out the differential metabolite. Metabolite identification and construction of related metabolic pathways and metabolic networks were achieved by using online databases such as HMDB and METLIN. The results showed that 12 differential metabolites in the serum of mice irradiated at 6.5 Gy that were associated with the radiation injury model, including lactic acid, alanine, urea, serine, threonine, glycerol, L-5-oxoproline, L-lysine, stearic acid, stearic acid, oleic acid, and 1-monopalmitoylglucoside. Two metabolic pathways were enriched: glycerolipid metabolism and metabolism of glycine, serine, and threonine. 18 differential metabolites in the serum of mice irradiated at 8.5 Gy were associated with the radiation injury model, including lactic acid, alanine, urea, L-leucine, glycerol, nonanoic acid, serine, threonine, L-5-oxoproline, phenylalanine, L-ornithine, 1,5-dehydroorbital, L-lysine, L-tyrosine, pectic, oleic, stearic, and cholesterol. Four metabolic pathways were enriched: phenylalanine, tyrosine, and tryptophan synthesis, phenylalanine metabolism, glyceride metabolism, and glycine, serine, and threonine metabolism. It was suggested that H. erinaceus polysaccharides could intervene in radiation injury by altering amino acid and fatty acid synthesis in mice. It was assumed that H. erinaceus polysaccharides regulated the level of metabolic pathways through lipid metabolism and amino acid metabolism, thus affecting energy metabolism and amino acid metabolism and exerting its therapeutic effect on radiation damage.
Animals ; Mice ; Metabolomics/methods* ; Gas Chromatography-Mass Spectrometry/methods* ; Polysaccharides/pharmacology* ; Male ; Hericium/chemistry* ; Drugs, Chinese Herbal/administration & dosage* ; Metabolome/drug effects* ; Gamma Rays/adverse effects*

Objective To screen the targets related to the metabolic enzymes involved in the cholesterol synthesis pathway that inhibits the polarization of macrophages towards M1 phenotype,and verify the intervention effects and underlying mechanisms in colorectal cancer cells.Methods Mouse colorectal cancer MC38 cells were divided into control group(si-NC)and experimental groups(the expression of enzymes in cholesterol synthesis pathway was interfered with siRNA for corresponding targets).RT-qPCR was used to detect the mRNA levels of corresponding targets in MC38 cells after transfection.After peritoneal macrophages were extracted from male C57BL/6 mice(6 weeks old,weighing 13～18 g),the macrophages were then treated with the conditioned media of MC38 cells transfected with different siRNAs for 48 h.RT-qPCR was employed to detect the mRNA levels of IL-1β,IL-6 and TNF-α in the macrophages so as to evaluate the effect of the culture media on the M1 polarization.MC38 cells were divided into control groups(OE-NC and sh-NC),farnesyl-diphosphate farnesyltransferase 1(FDFT1)overexpression group(OE-FDFT1)and FDFT1 knockdown group(sh-FDFT1).RT-qPCR was applied to detect the mRNA expression of FDFT1,and Western blotting was conducted to measure the protein level of FDFT1.C57BL/6 mice were subjected randomly to construct a subcutaneous tumor-bearing model and a model of intraperitoneal metastatic tumor(n=5)respectively.The growth of tumor mass was then measured.Flow cytometry was used to observe the proliferation and apoptosis of MC38 cells,and Trans well assay to detect migration ability of MC38 cells.Five C57BL/6 macrophage-depleted mice(established with injection of clodronate liposome suspension through tail vein)received intraperitoneal implantation to construct a metastasis model,and then the obtained tumor masses were then weighted.Results Compared with MC38 cells after si-NC transfection,the mRNA levels of corresponding targets in MC38 cells in the experimental groups were significantly reduced(P＜0.05).Significant increases were found in the mRNA levels of IL-1β,IL-6 and TNF-α of the macrophages with FDFT1 interference than the control cells(P＜0.05).There were no statistical differences in the proliferation,apoptosis and migration of MC38 cells in the control group(OE-NC and sh-NC)and the cells of the FDFT1 overexpression group and FDFT1 knockdown group(P＞0.05).In both the subcutaneous tumor-bearing model and the model of intraperitoneal metastatic tumor,the mass weight was significantly heavier in the OE-FDFT1 group than the OE-NC group(P＜0.01),and was notably smaller in the sh-FDFT1 group than the sh-NC group(P＜0.01).For the macrophage-depleted mouse tumor model,no remarkable change was observed in the tumor weight between the OE-FDFT1 group and the OE-NC group as well as the sh-FDFT1 group and the sh-NC group.Conclusion FDFT1,the metabolic enzyme in the cholesterol synthesis pathway of colorectal cancer tumor cells,is a potential target for tumor immunotherapy targeting macrophages,which promotes tumor progression by regulating macrophages.

Acute ischemic stroke is the most common type of stroke, characterized by high mortality, disability, and recurrence rates. Intravenous thrombolysis is the core treatment method. Among them, alteplase is effective as the standard drug, but it has limitations such as narrow time window, high risk of bleeding, and the need for continuous infusion. The new generation of improved drug tenecteplase has the advantages of single intravenous injection and higher fibrin specificity, making it a potential alternative drug to alteplase. In addition, endovascular therapy compensates for the low recanalization rate of large vessel occlusion on the basis of intravenous thrombolysis. This article reviews the clinical progress of teneplase in the treatment of acute ischemic stroke, aiming to provide reference for optimizing the treatment plan of acute ischemic stroke.

Objective To investigate the effect of miR-185-5p-mediated targeted negative regulation of transmembrane 9 superfamily member 1(TM9SF1)on proliferation,migration and autophagy in lung adenocarcinoma cells.Methods The expression of miR-185-5p in lung adenocarcinoma tissues was analyzed using dataset GSE51853 downloaded from the Gene Expression Omnibus(GEO)database.Potential target proteins of miR-185-5p were predicted using online databases(miRTargetLink,miRTarbase,and DIANA-microT-CD),and autophagy-related proteins were obtained from HADb.The intersected results from these four databases was identified,and survival curves of vascular endothelial growth factor A(VEGFA)and TM9SF1 within the overlapping candidates were analyzed using the StarBase database.TM9SF1 3'UTR wild-type(WT)or TM9SF1 3'UTR mutant(MUT)reporter plasmids were separately co-transfected with miR-185-5p control plasmid(CON)or miR-185-5p overexpression plasmid(over-miR-185-5p)into HEK-293T cells.A dual-luciferase reporter gene assay was employed to assess the binding interaction between miR-185-5p and TM9SF1 and quantify the subsequent luciferase activity.Western blotting was used to assess TM9SF1 protein expression levels in A549 cells transfected with over-miR-185-5p.A549 cells were divided into three groups:(1)CON+NC group,co-transfected with miR-185-5p control plasmid and TM9SF1 control plasmid;(2)over-miR-185-5p+NC group,co-transfected with over-miR-185-5p and TM9SF1 control plasmid;(3)over-miR-185-5p+over-TM9SF1 group,co-transfected with both miR-185-5p and TM9SF1 overexpression plasmids.EdU cell proliferation assay,wound healing assay,and Transwell migration assay were performed to validate the effects of miR-185-5p targeted binding to TM9SF1 on proliferation and migration capacities in lung adenocarcinoma.Changes in autophagic flux and mitochondrial membrane potential(MMP)of lung adenocarcinoma cells were detected using stubRFP-sensGFP-LC3 lentivirus and JC-1 assays,respectively.Results In the GSE51853 dataset,miR-185-5p expression level was significantly lower in lung adenocarcinoma tissues compared with normal lung tissues(P<0.01).qRT-PCR analysis revealed that miR-185-5p expression was downregulated in lung adenocarcinoma cell lines NCI-H1299 and A549 compared with normal lung epithelial cells BEAS-2B(P<0.01).Bioinformatics predictions using miRTargetLink,miRTarbase,DIANA-microT-CD,and HADb databases indicated that miR-185-5p could target and regulate the autophagy-related protein TM9SF1.Dual-luciferase reporter assays and Western blotting demonstrated that miR-185-5p directly bound to the 3'UTR region of TM9SF1 mRNA,and overexpression of miR-185-5p significantly reduced the expression of target protein TM9SF1(P<0.05).EdU cell proliferation,wound healing,and Transwell migration assays demonstrated that miR-185-5p overexpression inhibited proliferation and migration capacities of lung adenocarcinoma cells,whereas TM9SF1 overexpression could attenuate this inhibition effect(P<0.05).Results of stubRFP-sensGFP-LC3 for autophagic flux analysis demonstrated that overexpression of miR-185-5p enhanced autophagic flux in A549 cells,whereas co-overexpression of miR-185-5p and TM9SF1 suppressed autophagic flux.JC-1 assays showed a decreased MMP level in A549 cells after miR-185-5p overexpression,with higher MMP level observed when miR-185-5p and TM9SF1 were co-overexpressed.Conclusion miR-185-5p may suppress proliferation,migration,and autophagy capacities in lung adenocarcinoma cells by targeting TM9SF1 through negative regulation.

Objective:To explore the function of MDM2 and its relationship with p53 at the cellular level during H 2O 2 induced oxidative damage. Methods:MLE-12 HALI cell models were established using 0.5 mmol/L H 2O 2, and were divided into three groups: normal control group, H 2O 2 injury group, H 2O 2+MDM2 overexpressed group, and H 2O 2+MDM2 shRNA group. Infection of MLE-12 cells with adenovirus vector overexpressing and silencing MDM2; Using immunoprecipitation (Co-IP) to analyze the interaction between MDM2 and p53; Western blotting was used to detect the protein expression levels of MDM2, p53, Bcl-2, Bax, and cleared caspase-3 after HALI modeling; Measure the apoptosis rate of cells in each group. Results:After transcriptome sequencing,the p53 signaling pathway closely related to HALI. Compared with the normal group, the expression of MDM2 in the H 2O 2 injury group was lower ( P<0.05); Compared with the H 2O 2 injury group, overexpression of MDM2 resulted in a decrease in the apoptosis rate of MLE-12 cells ( P<0.05), a decrease in the expression levels of p53, Bax, and cleared caspase-3 proteins, and an upregulation of MDM2 and Bcl-2 protein expression ( P<0.05). Compared with the H 2O 2 injury group, when MDM2 was silenced, the cell apoptosis rate increased ( P<0.05), and the expression levels of p53, Bax, and cleared caspase-3 proteins were upregulated, while the expression levels of MDM2 and Bcl-2 proteins decreased ( P<0.05). Co-IP experiments showed that MDM2 binds to p53 protein. Conclusions:MDM2 can exert a protective effect on HALI by inhibiting MLE-12 cell apoptosis through the p53/Bcl-2/Bax axis.

Objective To investigate the effect of orthopedic distractor assisted reduction and percutaneous plate implantation in the treatment of distal tibial fractures.Methods 120 patients with distal tibial fractures treated in Wendeng orthopedic hospital of Shandong Province from January 2019 to January 2021 were randomly divided into two groups:the control group(60 cases,percutaneous plate implantation and internal fixation under traditional manual reduction)and the observation group(60 cases,orthopedic distractor assisted manual reduction and percutaneous plate implantation and internal fixation).The treatment effect,intraoperative indexes and postoperative fracture recovery of the two groups were compared,and the complications of the two groups were recorded and compared.Results After treatment,there was no significant difference in the excellent and good rate of fracture healing in the observation group(P＞0.05);The operation time[(72.56±27.54)min]and fracture healing time[(16.45±4.59)w]of the observation group were shorter than those of the control group[(89.94±28.20)min and(21.15±4.54)w],and the amount of intraoperative bleeding in the observation group[(82.27±20.14)ml]was less than that of the control group[(90.12±21.48)ml],the differences were statistically significant(P＜0.05);12 months after operation,the knee joint score of American Special Surgery Hospital(HSS)and ankle hindfoot score of American ankle Orthopedic Association(AOFAS)in the observation group[(85.24±7.52)and(84.58±7.29)]were lower than those in the control group[(74.45±7.64)and(74.56±6.38)](P＜0.05).There was no significant difference in the total incidence of complications between the two groups(P＞0.05).Conclusion Orthopedic distractor assisted reduction and percutaneous steel plate implantation and internal fixation have good curative effect,which can effectively reduce the amount of intraoperative bleeding,shorten the operation time,and have a good recovery of foot and ankle function after operation.

italic>Artemisia argyi is a traditional Chinese medicine in China, which is used as medicine with its leaves. The leaves of A. argyi mainly contain flavonoids, phenolic acids, volatile oils and other compounds, and have a variety of pharmacological activities. AP2/ERF transcription factors are abundant in plants and are mainly involved in plant growth and development, abiotic stress response and secondary metabolite biosynthesis regulation. However, there are few reports on the AP2/ERF gene family and its functions in A. argyi. In this study, we systematically identified the AP2/ERF gene family in A. argyi genome, and analyzed its phylogenetic tree, protein physicochemical properties, subcellular localization, conserved motifs, promoter elements, and expression patterns. The results showed that a total of 204 AP2/ERF transcription factors were identified in A. argyi genome, encoding proteins consisting of 88-483 amino acids with a relative molecular mass of 10-52.94 kDa and a theoretical isoelectric point of 4.62-9.88. Subcellular prediction showed that the majority of AP2/ERFs were located in the nucleus, cytoplasm, and the minority of them is located in the membrane and chloroplasts. According to the Arabidopsis AP2/ERF family classification, A. argyi AP2/ERF proteins were divided into four subfamilies: Soloist, AP2, ERF (B3, B4, B5, B6), and DREB (A1, A2, A4, A5, A6), among which DREB family accounted for the largest proportion, and the same subfamily had similar conserved motifs. cis-Acting element analysis showed that AP2/ERF promoters have a large number of elements responding to light and abiotic stress. Expression pattern analysis showed that most of the genes of the AP2/ERF family were dominantly expressed mainly in roots and stems, of which 19 were dominantly expressed in leaves, 77 would be induced to be expressed by methyl jasmonate, of which 16 genes were both dominantly expressed in leaves and induced to be expressed by methyl jasmonate, and these AP2/ERF genes may be the key regulatory genes for the synthesis of active ingredients in A. argyi leaves.This study lays the foundation for the functional study of AP2/ERF family genes and their regulating roles in the active components biosynthesis in A. argyi leaves.

OBJECTIVE To establish a high-performance liquid chromatography method for the simultaneous determination of 14 components including geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and praeruptorin B in Qingfei Yihuo tablets. METHODS Titank C18 (250 mm×4.6 mm, 5 μm) column was used; 0.1% phosphoric acid solution(A)-acetonitrile(B) was used as the mobile phase with gradient elution; detection wavelengths: geniposide at 238 nm, aloe-emodin, rhein, emodin, chrysophanol, physcion and mangiferin at 254 nm, berberine hydrochloride at 265 nm, baicalin, wogonoside, baicalein, and wogonin at 280 nm, praeruptorin A and praeruptorin B at 321 nm. The flow rate was 1.0 mL·min−1 and the column temperature was 35 ℃; the injection volume was 10 μL. RESULTS The linear ranges of geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein,aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and, praeruptorin B were 4.96−223.17, 0.84−42.22, 18.76−938.16, 4.46−223.17, 4.86−243.10, 1.59−79.32, 0.76−38.17, 1.03−51.49, 1.59−79.40, 1.21−60.72, 1.80−90.06, 0.91−45.48, 1.04−51.83 and 0.86−43.23 μg·mL−1, with r all ≥ 0.999 9. The RSDs of instrument precision, stability and reproducibility tests were <3%, and the average recoveries in sample(n=6) were >90%, with the RSDs <3%. CONCLUSION The method is simple and reproducible and can provide a scientific basis for improving the quality standard of Qingfei Yihuo tablets.