Objective:To investigate the dynamic changes of serum long non-coding ribonucleic acid antisense non-coding RNA in the INK4 locus (LncRNA ANRIL), long non-coding ribonucleic acid maternally expressed gene 3 (LncRNA MEG3) and inflammatory factors [interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-18 (IL-18) and interleukin-1β (IL-1β)] in patients with acute ischemic stroke and their correlation with neurological impairment.Methods:This prospective study included fifty-two patients with acute ischemic stroke admitted to the Central Hospital of Dalian University of Technology were included, and the neurological impairments of the patients were scored using the National Institutes of Health stroke scale (NIHSS) on the first day of the onset of the disease, and serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factors (IFN-γ, IL-6, IL-18 and IL-1β) were detected using enzyme-linked immunosorbent assay (ELISA) at the first, second and seventh day of the onset of the disease. The correlation between serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factor levels and neurological impairment at each time point was analyzed by Spearman correlation analysis.Results:Among 52 patients, mild group had 22 cases (NIHSS score <5 scores), and moderate-to-severe group had 30 cases (NIHSS score ≥5 scores). In acute ischemic stroke patients with moderate-to-severe group, the expression level of LncRNA MEG3 was higher than in the mild group, but the intergroup difference did not reach statistical significance ( P>0.05). LncRNA ANRIL, LncRNA MEG3, IFN-γ, IL-18 and IL-1β peaked at the same time point in acute ischemic stroke patients, all within second day of the onset of the disease. Spearman correlation analysis showed that the serum LncRNA MEG3 level on the first day of the onset of the disease was significantly positively correlated with NIHSS score ( P = 0.006), and serum LncRNA ANRIL level on seventh day was significantly positively correlated with NIHSS score ( P = 0.049). Conclusions:Serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factors (IL-18, IL-1β and IFN-γ) are highly expressed in patients with acute ischemic stroke, peaking on the second day of onset. LncRNA MEG3 levels on the first day of onset and LncRNA ANRIL levels on the seventh day of onset are helpful in determining the degree of neurological impairment in patients with acute ischemic stroke.

Objective:To investigate the dynamic changes of serum long non-coding ribonucleic acid antisense non-coding RNA in the INK4 locus (LncRNA ANRIL), long non-coding ribonucleic acid maternally expressed gene 3 (LncRNA MEG3) and inflammatory factors [interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-18 (IL-18) and interleukin-1β (IL-1β)] in patients with acute ischemic stroke and their correlation with neurological impairment.Methods:This prospective study included fifty-two patients with acute ischemic stroke admitted to the Central Hospital of Dalian University of Technology were included, and the neurological impairments of the patients were scored using the National Institutes of Health stroke scale (NIHSS) on the first day of the onset of the disease, and serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factors (IFN-γ, IL-6, IL-18 and IL-1β) were detected using enzyme-linked immunosorbent assay (ELISA) at the first, second and seventh day of the onset of the disease. The correlation between serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factor levels and neurological impairment at each time point was analyzed by Spearman correlation analysis.Results:Among 52 patients, mild group had 22 cases (NIHSS score <5 scores), and moderate-to-severe group had 30 cases (NIHSS score ≥5 scores). In acute ischemic stroke patients with moderate-to-severe group, the expression level of LncRNA MEG3 was higher than in the mild group, but the intergroup difference did not reach statistical significance ( P>0.05). LncRNA ANRIL, LncRNA MEG3, IFN-γ, IL-18 and IL-1β peaked at the same time point in acute ischemic stroke patients, all within second day of the onset of the disease. Spearman correlation analysis showed that the serum LncRNA MEG3 level on the first day of the onset of the disease was significantly positively correlated with NIHSS score ( P = 0.006), and serum LncRNA ANRIL level on seventh day was significantly positively correlated with NIHSS score ( P = 0.049). Conclusions:Serum LncRNA ANRIL, LncRNA MEG3 and inflammatory factors (IL-18, IL-1β and IFN-γ) are highly expressed in patients with acute ischemic stroke, peaking on the second day of onset. LncRNA MEG3 levels on the first day of onset and LncRNA ANRIL levels on the seventh day of onset are helpful in determining the degree of neurological impairment in patients with acute ischemic stroke.

This study investigated the full-genome molecular characteristics of a rotavirus vaccine-derived strain,G1P[8]geno-type A group rotavirus RVA/Human-wt/CHN/HN1140/2021/G1P[8](referred to as HN1140).The gene fragments of the HN1140 strain were amplified with reverse transcription-polymerase chain reaction(RT-PCR)combined with whole-genome primers to obtain the full genome sequence.Genotyping was performed with the online genotyping tool RotaC 2.0,and similarity and genetic evolution analyses for each gene segment were conducted in DNAstar5.1 and MEGA11.0 software.The genotype of the HN1140 strain was deter-mined to be G1-P[8]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis demonstrated that all 11 genomic segments clus-tered closely with the RotaTeq vaccine strains,sharing 99.7%-100%nucleotide sequence similarity.Notably,VP1,VP2,VP6,and NSP2-NSP5 segments showed 100%nucleotide identity with RotaTeq strains.Comparative genomic analysis identified 13 nucleotide and 8 amino acid substitutions between HN1140 and RotaTeq strains,localized within the VP7,VP4,VP1,VP2,VP3,and NSP1 segments.The HN1140 strain exhibited the genotype G1-P[8]-A3-T6-H3,which was consistent with the typical profile of a vaccine-derived reassortant.This strain demonstrated high genetic similarity to RotaTeq vaccine strains,with nucleotide sequence identity ranging from 99.7%to 100%.These findings suggested that HN1140 evolved from RotaTeq vaccine strains through genetic reassortment.

Methodological quality assessment is a pivotal link between primary studies and reliable evidence-based practice,and an essential pathway for operationalizing the core principles of the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition).A prevalent challenge in practice,however,is the conflation of appraising methodological robustness(risk of bias assessment)with verifying reporting transparency(adherence to reporting guidelines).This paper systematically addresses this fundamental challenge,beginning with a clear distinction between the essence and boundaries of these two concepts.On this basis,the article provides a comprehensive review of mainstream quality assessment tools,covering the methodological features and evolutionary trajectory of numerous instruments for interventional(e.g.,RoB 2,ROBINS-I),observational(e.g.,NOS,the JBI/SIGN/NIH series),secondary(e.g.,AMSTAR 2),and other specific types of studies such as health economic evaluations.Furthermore,a complete case study is used to illustrate the practical application of the ROBINS-I tool.The paper's central thesis advocates for an"appraisal-informed design"philosophy,urging a conceptual shift from the retrospective critique of existing literature to the prospective quality control of new research by internalizing appraisal standards as design principles,while also exploring the emerging paradigm of artificial intelligence in assisting assessment.This paper provides a comprehensive methodological reference for researchers and practitioners to prudently select appropriate assessment tools and to conduct rigorous critical appraisals of pharmacoepidemiological evidence.

Methodological quality assessment is a pivotal link between primary studies and reliable evidence-based practice,and an essential pathway for operationalizing the core principles of the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition).A prevalent challenge in practice,however,is the conflation of appraising methodological robustness(risk of bias assessment)with verifying reporting transparency(adherence to reporting guidelines).This paper systematically addresses this fundamental challenge,beginning with a clear distinction between the essence and boundaries of these two concepts.On this basis,the article provides a comprehensive review of mainstream quality assessment tools,covering the methodological features and evolutionary trajectory of numerous instruments for interventional(e.g.,RoB 2,ROBINS-I),observational(e.g.,NOS,the JBI/SIGN/NIH series),secondary(e.g.,AMSTAR 2),and other specific types of studies such as health economic evaluations.Furthermore,a complete case study is used to illustrate the practical application of the ROBINS-I tool.The paper's central thesis advocates for an"appraisal-informed design"philosophy,urging a conceptual shift from the retrospective critique of existing literature to the prospective quality control of new research by internalizing appraisal standards as design principles,while also exploring the emerging paradigm of artificial intelligence in assisting assessment.This paper provides a comprehensive methodological reference for researchers and practitioners to prudently select appropriate assessment tools and to conduct rigorous critical appraisals of pharmacoepidemiological evidence.

Objective:To analyze the carriage status, subtype distribution and flanking gene sequence characteristics of oxacillinases (OXA enzyme) in 241 clinical strains of Pseudomonas aeruginosa, and assess their roles in the drug resistance of Pseudomonas aeruginosa and ability to horizontally transfer across species. Methods:Clinical P. aeruginosa isolates were collected from four hospitals in Sanya, Tangshan, Zhangjiakou, and Beijing. The prevalence of oxacillinases and their flanking gene sequences was analyzed by whole-genome sequencing (NGS) and bioinformatic approaches. Results:A total of 241 isolates of P. aeruginosa were gathered, and 35 blaOXA subtypes were identified through screening of 252 blaOXA genes. These genes were classified into three subfamilies: blaOXA-50-like (241, 95.6%), blaOXA-1-like (9, 3.6%) and blaOXA-10-like (2, 0.8%). Among these, 11 subtypes (11, 31.4%) were novel blaOXA subtypes. Nine of these belonged to the blaOXA-50-like subfamily and were designated as blaOXA-1244, blaOXA-1245, blaOXA-1246, blaOXA-1250, blaOXA-1252, blaOXA-1253, blaOXA-1254, blaOXA-1255, and blaOXA-1256. The remaining two belonged to the blaOXA-10-like subfamily and were named blaOXA-1247 and blaOXA-1248. Compared to the amino acid sequence of OXA-10, the newly identified subtype OXA-1247 exhibited a mutation at position 117, where a valine was replaced by a leucine. This change was thought to improve the enzyme′s ability to hydrolyze carbapenems. In the analysis of the flanking sequences of the blaOXA genes, Class I integrons were identified in four bacterial strains. The variable regions of these integrons carried three distinct patterns of resistance gene cassettes: aac( 6′) -Ib-blaOXA-1247-ant( 3′′) -Ia, aac( 6′) -Ib-blaOXA-1248 and aac( 6′) -Ib- blaIMP-45-blaOXA-1-catB3. Among these, the strain BJ2326 carried a class I integron that was connected to the downstream IS CR1 element to form a composite class I integron structure, additionally carrying the resistance gene blaPER-1. Out of the 223 non-wild-type P. aeruginosa strains, 127 strains exhibited non-wild-type profiles to the four beta-lactam antibiotics MEM, CAZ, FEP, and TZP, with the combination of MEM+CAZ+FEP being the most prevalent, representing 57.0% of the total. Conclusions:The blaOXA genes in 241 clinical P. aeruginosa strains showed diversity. Some blaOXA genes had a co-transfer risk with the metallo-β-lactamase resistance gene blaIMP-45. Among the 11 newly discovered blaOXA subtypes, the new subtype OXA-1247 may have carbapenemase activity and potential for horizontal transfer.

This study investigated the full-genome molecular characteristics of a rotavirus vaccine-derived strain,G1P[8]geno-type A group rotavirus RVA/Human-wt/CHN/HN1140/2021/G1P[8](referred to as HN1140).The gene fragments of the HN1140 strain were amplified with reverse transcription-polymerase chain reaction(RT-PCR)combined with whole-genome primers to obtain the full genome sequence.Genotyping was performed with the online genotyping tool RotaC 2.0,and similarity and genetic evolution analyses for each gene segment were conducted in DNAstar5.1 and MEGA11.0 software.The genotype of the HN1140 strain was deter-mined to be G1-P[8]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis demonstrated that all 11 genomic segments clus-tered closely with the RotaTeq vaccine strains,sharing 99.7%-100%nucleotide sequence similarity.Notably,VP1,VP2,VP6,and NSP2-NSP5 segments showed 100%nucleotide identity with RotaTeq strains.Comparative genomic analysis identified 13 nucleotide and 8 amino acid substitutions between HN1140 and RotaTeq strains,localized within the VP7,VP4,VP1,VP2,VP3,and NSP1 segments.The HN1140 strain exhibited the genotype G1-P[8]-A3-T6-H3,which was consistent with the typical profile of a vaccine-derived reassortant.This strain demonstrated high genetic similarity to RotaTeq vaccine strains,with nucleotide sequence identity ranging from 99.7%to 100%.These findings suggested that HN1140 evolved from RotaTeq vaccine strains through genetic reassortment.

Objective To explore the technical process and clinical application of laparoscopic combined upper midline incision minimally invasive liver transplantation. Methods A retrospective analysis was conducted on 30 cases of laparoscopic combined upper midline incision minimally invasive liver transplantation. The cases were divided into cirrhosis group (15 cases) and liver failure group (15 cases) based on the primary disease. The surgical and postoperative conditions of the two groups were compared. Results All patients successfully underwent laparoscopic "clockwise" liver resection, with no cases of passive conversion to open surgery or intolerance to pneumoperitoneum. In 6 cases, the right lobe was relatively large, and the right hepatic ligaments could not be completely mobilized. One case required an additional reverse "L" incision during open surgery. All patients successfully completed the liver transplantation, with no major intraoperative bleeding, cardiovascular events, or other occurrences in the 30 patients. The model for end-stage liver disease (MELD) score in the cirrhosis group was lower than that in the liver failure group (P<0.001). There were no statistically significant differences between the two groups in terms of age, surgical time, blood loss, anhepatic phase, or cold ischemia time (all P>0.05). During the perioperative period, there was 1 case of hepatic artery embolism, 1 case of portal vein anastomotic stenosis, no complications of hepatic vein and inferior vena cava, and 3 cases of biliary anastomotic stenosis, all of which occurred in the liver failure group. Conclusions In strictly selected cases, the minimally invasive liver transplantation technique combining laparoscopic hepatectomy with upper midline incision for graft implantation has the advantages of smaller incisions, less bleeding, relatively easier operation, and faster postoperative recovery, which is worthy of clinical promotion and application.

Objective To explore the potential prognostic factors of patients with acute ischemic stroke(AIS)after undergoing endovascular mechanical thrombectomy and to construct an effective predictive model.Methods A retrospective analysis of clinical data was conducted on 202 patients with anterior circulation large vessel occlusion AIS from 2 stroke centers.All patients received endovascular mechanical thrombectomy treatment,with treatment and follow-up lasting at least 90 d.Basic demographic characteristics,medical records,and baseline blood biomarker data were collected,and the potential prognostic indicators for AIS after 90 d were screened using least absolute shrinkage and selection operator(LASSO)-logistic regression analysis.Results It was found that alcohol drinking(P=0.029),hypertension(P=0.001),diabetes mellitus(P=0.021),stroke or transient ischemic attack(P=0.049),systolic blood pressure on admission(P=0.009),diastolic blood pressure on admission(P=0.038),blood glucose(P=0.003),white blood cell count(P=0.001),neutrophil count(P=0.001),fibrinogen(P=0.010),systemic immune-inflammation index(P=0.008)and neutrophil-to-lymphocyte ratio(NLR)(P＜0.001)were associated with adverse clinical outcomes.Nine significant prognostic determinants were screened through LASSO-logistic regression analysis.Multivariate logistic regression analysis revealed that male sex(P=0.008),smoking history(P=0.013),hypertension(P=0.011),lymphocyte(P=0.028),fibrinogen(P=0.016),and NLR(P＜0.001)were significant predictive factors for poor prognosis in AIS patients after endovascular thrombectomy treatment.The constructed prognostic model had an accuracy of 76.2％,a sensitivity of 78.2％,a specificity of 71.7％,and a positive predictive value of 86.7％.Conclusion The predictive model established in this study can assist clinicians in identifying high-risk patients with AIS who have undergone endovascular thrombectomy,and it provide guidance for formulating individualized treatment strategies.

Objective:To explore the clinical and pathological features and survival outcomes of patients with early-onset intrahepatic cholangiocarcinoma (EOICC).Methods:This is a multicenter, retrospective cohort study. Data of 1 160 intrahepatic cholangiocarcinoma patients undergoing radical resection in 14 tertiary Grade A hospitals in China from January 2010 to November 2021 were retrospectively collected. The cohort included 632 males and 528 females, aged( M (IQR)) 61 (14) years (range: 22 to 93 years). ICC aged ≤50 years at the time of diagnosis was defined as EOICC and >50 years as late-onset intrahepatic cholangiocarcinoma (LOICC). Of these, there were 247 cases in the EOICC group and 913 cases in the LOICC. The clinical and pathological characteristics of both groups were analyzed and compared using the independent sample t-test, Mann-Whitney U test or Kaplan-Meier method. Univariate and multivariate Cox regression models for patient outcomes were constructed and forest graphed. Results:Compared with the patients in the LOICC group, patients in the EOICC group had lower carcinoembryonic antigen levels (2.5(4.0) μg/L vs. 3.1(5.2)μg/L, U=124 899, P=0.009) and CA19-9 level (63.4(524.7)U/ml vs. 77.9(611.3)U/ml, U=120 320, P=0.013), higher levels of ALT (29(35)U/L vs. 24(26)U/L, U=101 214, P=0.013), a lower score of the Eastern US Cooperative Oncology Group (0 score patients: 54.7% vs. 44.1%, χ2=12.472, P=0.014), higher TNM stage ( χ2=11.807, P=0.038), and proportion of lymph node dissection (62.3% vs. 54.1%, χ2=5.355, P=0.021). Patients in the two groups in sex, first diagnosis symptoms, intrahepatic bile duct stone history, nail protein, albumin, total bilirubin, transaminase, liver function Child-Pugh grade, T stage, stage, N stage, preoperative laparoscopic exploration proportion, tumor diameter, vascular invasion proportion, differentiation, margin, intraoperative bleeding, postoperative complications, postoperative hospital days were no statistical significance (all P>0.05). Patients in the EOICC group had better outcomes than the LOICC group (median survival time: 29.7 months vs. 25.0 months, 3-year overall survival: 45.1% vs. 37.8%, P=0.027). Conclusion:EOICC patients are better than LOICC patients in carcinoembryonic antigen, CA19-9, ALT, physical strength status and TNM stage, and the long-term prognosis is also better than LOICC patients.