ObjectiveTo investigate the mechanism of Xiaozhi Qinggan decoction （XQD） in preventing and treating metabolic dysfunction-associated steatotic liver disease （MASLD） by regulating ferroptosis， network pharmacology， in vitro and in vivo experiments. MethodsIn the in vivo experiment， mouse MASLD models were established by high-fat diet （HFD） induction. The model mice were randomly assigned to a positive control group （silybin， 50 mg·kg^-1）， low-， medium- and high-dose XQD groups （4.725， 9.45， 18.9 g·kg^-1）， with a normal control group. After 4 weeks of modeling， mice except the normal group were administered intragastrically for 8 consecutive weeks. Liver function， serum lipid levels， hepatic histopathology， as well as the levels of malondialdehyde （MDA）， superoxide dismutase （SOD）， reduced glutathione （GSH） and oxidized glutathione （GSSG） and Fe²⁺ were detected. The mRNA and protein expression of p53， SLC7A11 and GPX4 were determined by quantitative Real-time quantitative polymerase chain reaction（Real-time PCR） and Western blot. In the network pharmacology analysis， active components and potential targets of XQD for MASLD were screened， followed by functional and pathway enrichment analyses， and molecular docking was performed to verify the target binding activity. In the in vitro experiment， the optimal concentration of XQD-containing serum was screened by cytotoxicity assay. HepG2 cells were transfected with ov-NC or ov-p53 plasmid， and a lipid accumulation model was induced by free fatty acid （FFA， 1.0 mmol·L^-1）. Cells were divided into a normal group， FFA model group， ov-NC+XQD （15%） group and ov-p53+XQD （15%） group. Intracellular Fe²⁺ level and lipid accumulation were evaluated， and the protein expression of p53， SLC7A11 and GPX4 was measured by Western blot. ResultsCompared with the normal group， the model group exhibited markedly elevated body weight， liver weight， liver index， fasting blood glucose， AUC of glucose tolerance test， serum liver function and blood lipid levels at week 12 （P<0.01）. Hepatic steatosis and inflammatory infiltration were observed by pathological staining. Additionally， hepatic levels of MDA， SOD and Fe²⁺ were increased （P<0.01）， while GSH， GSSG and the GSH/GSSG ratio were decreased （P<0.01）. The mRNA and protein expression of hepatic p53 was upregulated （P<0.01）， whereas the expression of SLC7A11 and GPX4 was downregulated （P<0.01）. Compared with the model group， the low- and medium-dose XQD groups showed significantly decreased body weight at week 12 （P<0.05）. The silybin group， together with the medium- and high-dose XQD groups， presented reduced liver weight and liver index （P<0.05）. Fasting blood glucose and the AUC of glucose tolerance test were lowered in all four treatment groups （P<0.05， P<0.01）. Pathological staining revealed alleviated hepatic steatosis and inflammation， accompanied by decreased serum liver function and blood lipid levels （P<0.05， P<0.01）. Moreover， hepatic MDA and SOD levels were markedly reduced， while GSH， GSSG and the GSH/GSSG ratio were significantly elevated （P<0.05， P<0.01）. Hepatic Fe²⁺ level was decreased （P<0.01）. The mRNA and protein expression of hepatic p53 was downregulated， and the expression of SLC7A11 and GPX4 was upregulated （P<0.05， P<0.01）. Network pharmacology analysis identified quercetin， kaempferol， luteolin， tanshinone IIA and isorhamnetin as the core active components of XQD， with p53 serving as the key target. Stable binding was verified between these active components and the p53 protein. The optimal concentration of XQD-containing serum in vitro was determined to be 15%. Compared with the normal group， the model group showed increased intracellular Fe²⁺ and lipid accumulation， significantly upregulated p53 protein expression （P<0.01）， and markedly downregulated SLC7A11 and GPX4 protein expression （P<0.01）. Compared with the model group， the ov-NC group exhibited reduced Fe²⁺ and lipid accumulation， downregulated p53 expression， and upregulated SLC7A11 and GPX4 expression. In the ov-p53 group， p53 expression was upregulated （P<0.01）， while SLC7A11 and GPX4 expression was downregulated （P<0.01）. ConclusionXQD inhibits ferroptosis by downregulating p53 and upregulating SLC7A11 and GPX4， thereby alleviating oxidative stress and lipid peroxidation in hepatocytes and improving MASLD.

It is believed that patients with cirrhotic ascites exhibit a pathological mechanism characterized by the decline of healthy qi and the accumulation of pathogenic factors. Clinically， treatment should be based on the theory of tonification and purging， with a staged approach distinguishing between the active phase and the remission phase. The balance between tonification and purging should be adjusted according to the progression of pathogenic and healthy actors. In the acute phase， purging should take precedence over tonification， using purging as a means of tonification to facilitate the flow of water and qi through the triple energizer. The severity of water retention， dampness， blood stasis， and heat should be carefully assessed to ensure thorough elimination of pathogenic factors while avoiding harm to healthy qi. Medication adjustments should be made once the pathogenic factors are significantly weakened. In the remission phase， an integrated approach combining both tonification and purging should be adopted， incorporating purging within tonification to clear residual pathogens and prevent recurrence. Concurrently， proactive treatment of the underlying disease is essential to achieve complete recovery and prevent the recurrence of ascites.

Objective To explore the safety of Yttrium-90 resin microsphere selective internal radiation therapy (⁹⁰Y-SIRT) on malignant liver tumors. Methods A retrospective analysis was conducted on 64 patients with malignant liver tumors who underwent ⁹⁰Y-SIRT from February 2023 to November 2024 at Weifang People’s Hospital. The clinical characteristics of the patients and the occurrence of adverse reactions after treatment were analyzed to assess the safety of ⁹⁰Y-SIRT. Results Among the 64 patients, there were 52 males (81.25%) and 12 females (18.75%); the average age was (56.29±11.08) years. Seven patients (10.94%) had tumors with maximum diameter of less than 5 cm, 38 patients (59.38%) had tumors with maximum diameter of 5-10 cm, and 19 patients (29.68%) had tumors with maximum diameter of greater than 10 cm. There were 47 cases (73.44%) of solitary lesions and 17 cases (26.56%) of multiple lesions; 53 cases (82.81%) were primary liver cancers and 11 cases (17.19%) were metastatic liver cancers. Of the 64 patients, 63 successfully completed the Technetium-99m macroaggregated albumin (^99mTc-MAA) perfusion test and received the ⁹⁰Y-SIRT; one patient received ⁹⁰Y-SIRT after the second ^99mTc-MAA perfusion test due to a work error. The most common adverse reactions included grade 1 alanine aminotransferase (ALT) elevation in 26 cases (40.62%) and grade 2 in 2 cases (9.37%), grade 1 aspartate aminotransferase (AST) elevation in 27 cases (42.18%) and grade 2 in 7 cases (10.93%); grade 1 nausea in 17 cases (26.56%) and grade 2 in 6 cases (9.37%); grade 1 abdominal pain in 12 cases (18.75%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%); grade 1 vomiting in 11 cases (17.18%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%). Conclusion The adverse reactions of ⁹⁰Y-SIRT for treating malignant liver tumors are mild, indicating good safety.

Objective To construct one high-copy BAC vector with cloning capacity for large DNA fragment insertion and use it for the haplotype analysis of CYP2A6 gene.Methods Oligos including multiple cloning sites were annealed and ligated into the Hind Ⅲ/BamH Ⅰ site of pGEM-3Z and pBeloBACll,separately.Then,the intermediate vectors were digested by Hind Ⅲ and ligated together,so as to get the head-to-tail oriented high-copy BAC vector pBAC-BJH after the blue-white spotting test.STI PCR method was used for the amplification of whole CYP2A6 gene,and purified amplicon was double digested by BstB Ⅰ/Mlu Ⅰ and ligated into the same site of newly constructed vector pBAC-BJH.Several clones were then picked up and sequenced for the haplotype analysis of carriers with newly discovered CYP2A6 variant 355A＞T.Results One high-copy BAC vector pBAC-BJH with 7 newly added multiple cloning sites was successfully constructed.High copy number and multiple cloning sites were the advantages of this plasmid.With this vector,haplotype analysis result for 22C＞T,51A＞G,355A＞T in carrier with newly detected CYP2A6mutation was CGT.Conclusion One vector pBAC-BJH convenient for cloning large DNA fragment was successfully developed,and it can be used for the haplotype analysis of cytochrome p450 gene and cloning or sequencing of other genes with large genomic DNA inserts.

AIM: To investigate the effect of the timing of satisfactory sedation with preoperative oral midazolam on anesthesia induction and recovery in children undergoing adenotonsillectomy. METHODS: A total of 147 children undergoing elective adenotonsillectomy, with ASA physical status orⅡ, aged 2-7 years were selected from November 2022 to June 2023 in the Second Affiliated Hospital of Wenzhou Medical University. The children were orally administered 0.5 mg/kg midazolam in preoperative waiting area and were divided into 10-20 min (rapid onset, M1 group) and 21-30 min (slow onset, M2 group) based on the satisfactory sedation time, or equal volume of sugar pear drink orally (blank control group, C group). Children in all three groups received a general anesthesia method of propofol+fentanyl combined with sevoflurane induction and sevoflurane maintenance. The primary outcome measures were the induction compliance checklist (ICC) score and the pediatric anesthesia emergence delirium (PAED) score in the post-anesthesia care unit (PACU) to assess the occurrence of emergence agitation (EA), and the secondary outcome measures included the parental separation anxiety scale (PSAS), sedation Ramsay score, surgery duration, recovery time, PACU stay time, discharge time, the incidence of perioperative respiratory adverse events (PRAE) and other adverse events in the ward. RESULTS: 147 children were included in the result analysis, with 49 cases in each group. The proportion of perfect induction (ICC=0) were significantly higher in two M groups than that in group C (95.9% vs. 91.8% vs. 61.2%, P=0.001). The maximum and average PAED score in PACU in group M1 showed a significantly higher (6.4±5.0 vs. 4.4 ± 4.1, P=0.029; 5.2 ± 4.5 vs. 3.4 ± 3.6, P=0.030), and the incidence of EA was significantly higher than those in group C (10.2% vs. 30.6%, P=0.022), and increased compared to the group M2 (OR= 0.581, 95%CI 0.231-1.463, P=0.354). There was no statistically significant difference in the maximum and average PAED scores, incidence of EA between group M2 and group C (P>0.05). The Ramsay score and PSAS score in two M groups were higher, PACU stay time and recovery time was longer than those in group C (P<0.05). The pain scores in PACU in group M1 was higher than that of group C (P<0.05). There was no statistically significant difference in the surgical time, discharge time, the incidence of PRAE and other adverse events in the ward among three groups (P>0.05). CONCLUSION: Preoperative oral midazolam can improve the ICC and PSAS scores of children during induction, but it also leads to prolonged recovery time and PACU retention time. The rapid onset of midazolam did not result in better induction and recovery quality, but instead increased the incidence of EA and postoperative pain score.

Objective To explore the status quo of medication belief in the patients with myasthenia gravis and analyze their influencing factors,so as to provide reference for health care professionals to develop targeted interventions.Methods A total of 145 patients with myasthenia gravis visiting the First Affiliated Hospital of Guangzhou University of Chinese Medicine from July 2021 to March 2022 were selected.The Be-liefs about Medicines Questionnaire(BMQ)was used to investigate.The multiple linear regression was used to analyze the relevant influencing factors.Results The scores of medication belief,necessity belief and con-cern belief in 145 patients were(4.17±1.23)points,(19.52±3.45)points and(18.29±4.26)points respec-tively.There was statistically significant difference between the scores of necessity belief and concern belief(P＜0.05).The education level,financial burden,duration of illness,length of medication,number of recur-rent hospitalizations,and inappropriate medication-induced exacerbations had influence on the medication be-lief scores of the patients with myasthenia gravis(P＜0.05).The duration of illness,length of medication and number of recurrent hospitalizations had the influence on the medication necessity scores of patients with my-asthenia gravis(P＜0.05).The financial burden had the influence on the medication concerns scores of the patients with myasthenia gravis(P＜0.05).Conclusion The medication belief in the patient swith myasthe-nia gravis is at a low level,and the number of recurrent hospitalizations and financial burden are the independ-ent risk factors affecting the medication belief scores in the patients with myasthenia gravis.The number of recurrent hospitalizations is an independent risk factor for the score of medication necessity dimension.

The fat mass and obesity-associated protein (FTO) is an RNA demethylase required for catalytic demethylation of N ⁶-methyladenosine (m⁶A); it is highly expressed and functions as an oncogene in acute myeloid leukemia (AML). Currently, the overarching objective of targeting FTO is to precisely inhibit the catalytic activity. Meanwhile, whether FTO degradation also exerts antileukemic effects remains unknown. Herein, we designed the first FTO-targeting proteolysis targeting chimera (PROTAC) degrader QP73 using our FTO inhibitor Dac85-which potently inhibits FTO demethylation in AML cell lines-as a warhead. Notably, QP73 significantly induced FTO degradation in a time-, dose-, and ubiquitin-proteasome system-dependent manner and had superior antiproliferative activities to the FTO inhibitor Dac85 in various AML cell lines. Moreover, QP73 treatment significantly increased m⁶A modification on mRNA, promoted myeloid differentiation, and induced apoptosis of AML cells. Quantitative proteomics analysis showed that QP73 induced complete FTO degradation, upregulating RARA and ASB2 abundance and downregulating CEBPA, MYC, PFKP, and LDHB levels in AML cells. Lastly, QP73 exhibited antileukemic activity by increasing m⁶A modification and decreasing FTO levels in xenograft AML tumors. This proof-of-concept study shows that FTO-targeting PROTAC degraders can regulate the FTO signaling pathway and have potential antileukemia applications.

Emerging SARS-CoV-2 variants have made COVID-19 convalescents susceptible to re-infection and have raised concern about the efficacy of inactivated vaccination in neutralization against emerging variants and antigen-specific B cell response. To this end, a study on a long-term cohort of 208 participants who have recovered from COVID-19 was conducted, and the participants were followed up at 3.3 (Visit 1), 9.2 (Visit 2), and 18.5 (Visit 3) months after SARS-CoV-2 infection. They were classified into three groups (no-vaccination (n = 54), one-dose (n = 62), and two-dose (n = 92) groups) on the basis of the administration of inactivated vaccination. The neutralizing antibody (NAb) titers against the wild-type virus continued to decrease in the no-vaccination group, but they rose significantly in the one-dose and two-dose groups, with the highest NAb titers being observed in the two-dose group at Visit 3. The NAb titers against the Delta variant for the no-vaccination, one-dose, and two-dose groups decreased by 3.3, 1.9, and 2.3 folds relative to the wild-type virus, respectively, and those against the Omicron variant decreased by 7.0, 4.0, and 3.8 folds, respectively. Similarly, the responses of SARS-CoV-2 RBD-specific B cells and memory B cells were boosted by the second vaccine dose. Results showed that the convalescents benefited from the administration of the inactivated vaccine (one or two doses), which enhanced neutralization against highly mutated SARS-CoV-2 variants and memory B cell responses. Two doses of inactivated vaccine among COVID-19 convalescents are therefore recommended for the prevention of the COVID-19 pandemic, and vaccination guidelines and policies need to be updated.

AIM: To evaluation the effects of esketamine and butorphanol on postoperative pruritus induced by epidural morphine injection in cesarean delivery parturients. METHODS: A total of 162 parturients who underwent elective cesarean section under continuous epidural anesthesia in Taizhou Central Hospital (Taizhou University Hospital), were selected and randomly divided into esketamine group (group K), butorphanol group (group B) and blank control group (group C). 5min after umbilicus amputation, parturients in group K was injected with 3 mg morphine diluent through epidural catheter, and esketamine 0.2 mg/kg intravenously. Parturients in groups B and C were given the same dose of morphine,and butorphanol 10 μg/kg or the same volume of normal saline, respectively. The incidence of postoperative pruritus at different times, the degree of pruritus and incidence of other adverse reactions were compared among three groups. RESULTS: The highest incidence of pruritus occurred within 4 hours after operation. The incidence of postoperative pruritus at 4 hours in group K and B was significantly lower than that in group C (3.7% vs. 3.7% vs. 29.6%, P < 0.05), the total incidence of postoperative pruritus within 48 hours was also significantly lower than that in group C (13.0% vs. 11.1% vs. 40.7%, P < 0.05), and the incidence of moderate to severe pruritus was also significantly lower than that of group C (5.6% vs. 3.7% vs. 31.5%, P < 0.05). There was no significant difference between group K and group B (all P > 0.05). There were no significant differences in the incidence of postoperative nausea, vomiting, dizziness and postoperative pain scores among three groups (P > 0.05). CONCLUSION: Both esketamine and butorphanol can reduce the incidence and degree of pruritus caused by epidural morphine injection in parturients, without affecting the analgesic effect of morphine and without increasing the incidence of adverse reactions. Esketamine is as effective and safe as butorphanol in preventing pruritus after cesarean section.

Objective:To assess the applicability of the Chinese version of the symptom assessment in dry eye (SANDE) questionnaire refer to the ocular surface disease index (OSDI) questionnaire.Methods:A cross-sectional study was conducted.Three hundred and twenty-three students from a senior high school in Hongkou District in Shanghai were enrolled in June 2020.The Chinese version of the SANDE and the OSDI questionnaires were answered by the students to assess the dry eye symptom and were collected on-site.Scores of the two questionnaires were calculated.According to the OSDI score, the students were divided into the normal control group ( n=87), mild dry eye group ( n=82), moderate dry eye group ( n=87) and severe dry eye group ( n=67). Cronbach α was obtained to evaluate the internal consistency.Kruskal-Wallis test was used to analyze the difference in SANDE scores among dry eye groups with different severities and evaluate the discriminative validity.Correlation between the total scores of the two questionnaires was analyzed by using Spearman rank correlation analysis to evaluate the criterion validity.Factor analysis was used to evaluate the construct validity.Receiver operating characteristic curve (ROC) was drawn to obtain the area under the ROC curve (AUC) and cut-off point to evaluate the diagnostic threshold for dry eye.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Shanghai General Hospital (No.2020KY026). Written informed consent was obtained from guardians of each subject. Results:The Cronbach α of the SANDE and OSDI questionnaires were 0.856 and 0.829, respectively.SANDE score of the four groups classified according to the OSDI score was 7.0 (2.5, 16.9) for the normal control group, 17.0 (8.4, 31.0) for the mild dry eye group, 29.2 (14.6, 49.4) for the moderate dry eye group and 49.1 (24.4, 60.7) for the severe dry eye group, respectively, which was increased in turn.There was a statistically significant overall difference in the SANDE score among the four groups ( H=113.213, P<0.001), and statistically significant pairuise differences were found (all at P<0.05). The SANDE score was moderately positively associated with the OSDI score ( rs=0.615, P<0.001). The factor analysis revealed that for the SANDE questionnaire, factor loadings of its two items on the common factors were higher than 0.5, among which the frequency of dry eye symptoms was 0.936, and the severity of dry eye symptoms was 0.936.The AUC of the SANDE questionnaire was 0.815 ( P<0.001, 95% CI: 0.770-0.860). When using 23 as the diagnostic threshold for dry eye, the sensitivity and specificity of the SANDE questionnaire were 60.59% and 88.51%, respectively. Conclusions:The Chinese version of the SANDE questionnaire shows good reliability, validity, discrimination ability and slightly poor sensitivity in detecting dry eye.As a simple assessment questionnaire, it is applicable to the Chinese population and is of good clinical application value.