Objective Objective To analyze the potential spatial factors affecting the genetic differentiation of Oncomelania hupensis robertsoni in Yunnan Province. Methods A total of 13 administrative villages were selected from schistosomiasis-endemic areas of Yunnan Province as O. hupensis snail sampling sites. At least 200 snails were collected in each site, and the spatial variable data of each site were recorded, including longitude, latitude and altitude. Thirty active and Schistosoma japonicum uninfected O. hupensis snails were selected from each sampling site by means of the crawling method and the cercarial shedding method. Genomic DNA was extracted from O. hupensis snails. Following PCR amplification, purification of PCR amplification products and sequencing, the gene sequences of O. hupensis snail samples were spliced and edited using the DNAstar software and the NCBI database to yield the complete mitochondrial sequences of O. hupensis snails at each sampling site, and the mitochondrial genetic distance matrix of O. hupensis robertsoni was calculated at each sampling site. The geographical coordinates of each sampling site were marked using the software ArcGIS 10.2, and the straight-line geographical distance between each sampling site was calculated. The altitude difference, longitude difference and latitude difference between each sampling site were calculated using the Excel software, and the correlation between the mitochondrial genetic distance matrix of O. hupensis robertsoni and each spatial variable matrix was examined by using the Mantel test at 13 sampling sites in Yunnan Province. Results Among the 13 O. hupensis snail sampling sites in Yunnan Province, the largest mitochondrial genetic distance of O. hupensis robertsoni snail populations was seen between Anding Village, Nanjian Yi Autonomous County and Caizhuang Village, Midu County (26.244 2), and the largest geographical distance was seen between Dongyuan Village, Gucheng District and Cangling Village, Chuxiong County (272.64 km). The highest altitude difference was seen between Anding Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1 086.10 m), and the largest longitude difference was found between Qiandian Village, Eryuan County and Cangling Village, Chuxiong County (1.86°), while the largest latitude difference was measured between Leqiu Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1.81°). In addition, the mitochondrial genetic distance of O. hupensis robertsoni snail populations was positively correlated with altitude at 13 snail sampling sites in Yunnan Province (r = 0.542 8, P < 0.001), and showed no significant correlations with geographical distance (r = 0.093 4, P > 0.05), longitude (r = −0.199 5, P > 0.05) or latitude (r = 0.205 7, P > 0.05). Conclusion Altitude may be a potential spatial factor affecting the genetic differentiation of O. hupensis robertsoni in Yunnan Province.

Objective To investigate the value of serum tumor markers in the diagnosis of lung cancer.Methods A total of 138 patients with newly diagnosed lung cancer patient(lung cancer group)and 52 patients with benign lung diseases(control group)in Anhui NO.2 Provincial People's Hospital from June 2020 to March 2024 were selected as the research objects.The clinical data and serum tumor markers[neuron-specific enolase(NSE),cytokeratin 19 fragment(CYFRA21-1),carcinoembryonic antigen(CEA),Pro-gastrin-relea-sing peptide(ProGRP)and squamous cell carcinoma antigen(SCC)]of all groups were compared.Results There were significant differences in age and smoking history between the two groups(P＜0.05).The serum levels of NSE,CYFRA21-1,CEA,ProGRP and SCC in lung cancer group were significantly higher than those in control group,and the differences were statistically significant(P＜0.05).The results of binary Logistic regression analysis showed that smoking history,age,CYFRA21-1 and CEA levels were all influen-cing factors for the occurrence of lung cancer(P＜0.05).Receiver operating characteristic(ROC)curve analy-sis results showed that the area under the curve(AUC)of smoking history,age,CEA and CYFRA21-1 in the combined diagnosis of lung cancer was 0.906(95％CI:0.865-0.947),which was higher than the AUC for the individual diagnosis of each index.The levels of serum CYFR21-1 and CEA in the stage Ⅰ—Ⅱ group were significantly lower than those in the stage Ⅲ-Ⅳ group,and the difference was statistically significant(P＜0.05).The results of binary Logistic regression analysis showed that CYFRA21-1 and CEA levels were influ-ential factors in the occurrence of lung cancer stage Ⅲ-Ⅳ(P＜0.05).ROC curve analysis showed that the AUC of CEA and CYFRA21-1 for the diagnosis of lung cancer stage Ⅲ-Ⅳ were 0.750(95％CI:0.667-0.832)and 0.771(95％CI:0.691-0.852),respectively.The AUC of combined diagnosis was 0.834(95％CI:0.765-0.902),the sensitivity was 58.9％,and the specificity was 95.3％.Conclusion Smoking history,age,CEA,and CYFRA21-1 may be potential biomarkers for the diagnosis and prognosis of lung cancer,espe-cially CEA and CYFRA21-1,which are also of great significance for the staging of lung cancer from stage Ⅲ to stage Ⅳ.

Objective: To investigate the protective effects of morusin on lipopolysaccharide (LPS)-induced acute liver injury in mice and its underlying mechanisms. Methods: Thirty-two male specific pathogen-free (SPF) C57BL/6J mice were randomly divided into four groups (n = 8 per group): control, LPS, low-dose morusin (morusin-L, 10 mg/kg), and high-dose morusin (morusin-H, 20 mg/kg) groups. The mice in each group were administered the corresponding drugs or normal saline via continuous gavage daily for 16 consecutive days. Except for control group, which received an equal volume of normal saline, other groups were intraperitoneally injected with LPS (5 mg/kg) 2 h after the last gavage to establish the acute liver injury model. Serum and liver tissues were collected for subsequent analysis 6 h after LPS injection. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected with biochemical methods. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in serum were measured by enzyme-linked immunosorbent assay (ELISA). Hepatic pathological changes were evaluated by hematoxylin-eosin (HE) staining. The 16S ribosomal RNA (16S rRNA) sequencing was performed to assess the composition of intestinal flora, linear discriminant analysis effect size (LEfSe) was applied for multi-level species discrimination, and Spearman’s correlation analysis was performed. The liver tissues of mice with acute liver injury were analyzed by RNA sequencing (RNA-seq) technology to identify differentially expressed genes (DEGs), and then enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was conducted. The expression levels of selected genes was validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR), while immunohistochemistry (IHC) was performed to examine the expression levels of IL-6, myeloid differentiation primary response 88 (MYD88), and toll-like receptor 2 (TLR2). Results: Morusin significantly reduced the serum levels of ALT, AST, and inflammatory factors (TNF-α, IL-6, and IL-1β) (P < 0.05, P < 0.01, or P < 0.001), while alleviating the hepatic pathological damage in mice. Based on efficacy comparisons, morusin-H group was selected for subsequent microbiome and transcriptome analyses. Microbiome analysis revealed that morusin-H effectively mitigated LPS-induced gut dysbiosis and restored the Firmicutes/Bacteroidota balance (P < 0.01). At the genus level, morusin-H significantly reduced the abundances of norank_f_Muribaculaceae, Desulfovibrio, Parabacteroides, and Muribaculum (P < 0.05, P < 0.01, or P < 0.001). At the phylum, family, and genus levels, our findings indicated that morusin-H treatment caused a significant decrease in the abundance of Desulfobacterota, Desulfovibrionaceae, and Desulfovibrio (P < 0.01). Importantly, the abundance of Desulfovibrio was positively correlated with the levels of ALT, AST, TNF-α, IL-1β, and IL-6. Transcriptomic and molecular analyses showed that the therapeutic mechanism of morusin-H involved suppression of the IL-17/TNF signaling pathways and downregulating the mRNA levels of Tlr2, Tlr3, Myd88, Il6, and Cxcl10 (P < 0.05 or P < 0.001), as well as the protein levels of key inflammatory mediators (IL-6, MYD88, and TLR2) (P < 0.001). Conclusion Morusin demonstrates protective effects against LPS-induced acute liver injury, likely through modulation of gut microbiota and suppression of pro-inflammatory factor expression. These findings indicate that morusin exerts its effects through the "microbiota-inflammation-liver" axis, providing a theoretical basis for its use as a multi-target plant-based drug in the treatment of metabolic inflammation-related liver diseases.

OBJECTIVE To compare the emergency specialist clinical pharmacist training programs between China and the United States， providing valuable insight for the development of specialized clinical pharmacist training in emergency departments within China. METHODS By reviewing the official website of the American Society of Health-System Pharmacists （ASHP）， the websites of some training institutions offering PGY2 emergency medicine （EM） residency programs in the United States， the official website of China’s National Health Commission， and the website of the Pharmaceutical Affairs Committee of the Chinese Hospital Association， relevant materials and data on the training of emergency medicine clinical pharmacists were collected. Microsoft Excel and NVivo software were utilized to analyze the implementation status of these training programs. Literature searches were conducted via Chinese （CNKI） and English （PubMed） databases， followed by screening， categorization， and thematic analysis aligned with research objectives. RESULTS As of now， there are 115 accredited PGY2 EM residency programs in the United States， which provide 120 specialized pharmacist training positions. These programs are distributed across 35 states and are hosted by a variety of institutions， including hospitals， medical centers， and universities. The predominant training model follows a hospital+acute care framework. Eligibility requirements for PGY2 EM residency programs include possession of a doctor of pharmacy （Pharm.D.） degree， pharmacist licensure， and completion of a PGY1 residency. The training standards are structured into three tiers： competency areas， competency goals， and learning objectives. The curriculum typically includes core rotations， elective rotations， and longitudinal training components. Assessment is conducted through a combination of formative and summative evaluations， with results categorized into four proficiency levels. In China， there is only one training base currently for emergency clinical pharmacist specialty training with an annual enrollment of three trainees. Applicant eligibility primarily involves requirements regarding academic degree， professional background， years of experience， and professional title. The training content covers four domains： general competency， clinical theoretical knowledge and skills， pharmacological knowledge and application， and clinical medication practice skills. The training process centers on rotations within emergency departments. Assessment methods include theoretical examinations， daily performance evaluations， and final completion assessments. CONCLUSIONS PGY2 EM residency programs in the United States emphasize inclusivity and professionalism in their implementation. Program admission involves a rigorous selection process， and they offer attractive incentive structures for trainees. The training content focuses on competency-based approaches and pragmatic applicability， while assessment methods are closely aligned with defined competence objectives. In contrast， specialist clinical pharmacist training in emergency medicine in China is currently in the exploratory and nascent stages. Admission criteria tend to be less stringent， and incentives for trainees are often insufficient. The training content appears relatively stereotyped and superficial， with assessment methods still primarily reliant on quantifiable metrics. In expanding and popularizing China’s emergency specialist clinical pharmacist training programs， it is essential to draw on advanced experiences from developed countries like the United States， particularly in areas such as training base distribution， application requirements， training content， and assessment methods. Aligned with the realities of emergency clinical practice in China， efforts should focus on enhancing program accessibility and training efficacy.

To establish an immunochromatographic method for rapid detection of caprine enterovir-us(CEV),the monoclonal antibody against CEV VP1 protein was used as gold-labeled monoclonal antibodies,and the purified rabbit-derived polyclonal antibody of CEV-VP1 and sheep anti-mouse IgG were used as the detection line and quality control line,respectively.The colloidal gold immu-nochromatographic test strips for CEV were prepared according to the principle of double antibody sandwich,evaluated,and applied for clinical specimen detection.The results showed that the meth-od specifically recognized CEV without cross-reaction with bovine enterovirus and bovine viral di-arrhea virus.The minimum detection limit of the method was 102.49 TCID50/mL and had good re-producibility.The prepared test strips had a shelf life of three months kept at 4 ℃.Detection of clin-ical samples using the immunochromatographic test strips showed 100％coincidence rate with RT-PCR method.In conclusion,the colloidal gold immunochromatographic test strips for detection of the emerging CEV with good specificity,sensitivity and repeatability,which provides a new techni-cal means easily used for the rapid detection/diagnosis and epidemiological investigation on CEV infection.

Based on the theory of the liver's"using bitter herbs to nourish or purge"from Huangdi Neijing,this paper systematically elucidates the theoretical foundation for treating functional constipation from liver.Focusing on the physiological characteristic of"liver desires to disperse"and the pathological manifestation of"liver bitterness and urgency,"combined with the"liver communicates with the large intestine"theory,this paper establishes a diagnostic and therapeutic framework for managing functional constipation by regulating liver function.The pathological evolution of functional constipation manifests in three distinct stages:in the early stage,liver qi stagnation leads to large intestine qi obstruction,where damaged by an excess of seven emotions resulting in symptoms such as difficult defecation,abdominal bloating,and hypochondriac pain;in the middle stage,liver depression transforms into fire,scorching bodily fluids to generate dryness,thereby creating a pathological interplay of stagnation,fire,and dryness,which is marked by anal heat,dry mouth,and yellow urine;in the late stage,yin deficiency in liver and kidney causes large intestine malnutrition,resulting in a complex pathological state where yin deficiency,collateral blockage,dryness accumulation,and blood stasis intertwine,clinically manifesting as pellet-like stools(resembling sheep feces)and soreness and weakness of the waist and knees.In treatment,the formula design follows the principle of"sweetness to relieve,acridity to tonify,and sourness to purge,"with treatment principles varying across stages.In the early stage,the focus is on dispersing liver and regulating qi,and unblocking the zang-fu viscera;in the middle stage,the priority shifts to clearing heat-fire,nourishing large intestine,and promoting fluid production;whereas,in the late stage,the emphasis lies on nourishing yin,unblocking collaterals,and promoting blood circulation.This staged treatment of functional constipation overcomes the limitations of solely focusing on nourishing large intestine and facilitating feces excretion,thereby advancing the treatment of different stages based on syndrome differentiation and personalized treatment.It provides theoretical support for improving patients' intestinal function and enhancing overall health outcomes.

AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.

Objective:To investigate the feasibility of transcatheter edge-to-edge repair (TEER) using a short-clip strategy for patients with moderate-to-severe or greater degenerative mitral regurgitation caused by commissural prolapse.Methods:This retrospective study included patients with severe mitral regurgitation secondary to commissural prolapse who underwent TEER at the Second Affiliated Hospital of Zhejiang University School of Medicine between September 2022 and July 2024. Preoperative clinical and imaging data, intraoperative details, procedural outcomes, and 1-month postoperative follow-up results were collected.Results:A total of 19 patients were enrolled, aged (74.1±6.1) years, including 12 males. Among them, 10 patients had external commissural prolapse, and 9 patients had internal commissural prolapse. Preoperatively, all patients exhibited severe mitral regurgitation (4+), with an effective regurgitant orifice area of (0.55±0.17) cm2, left atrial volume of (104.77±36.57) ml, left ventricular end-diastolic volume of (102.29±32.47) ml, left ventricular end-diastolic dimension of (5.34±0.59) mm, and prolapse width of (1.18±0.34) cm. All procedures utilized short clips (NTR or NTW clips) to target the prolapsed commissural region and were completed successfully without intraoperative complications. At 1-month follow-up, no mortality, stroke, single-leaflet device attachment, myocardial infarction, or unplanned mitral reintervention occurred. Mitral regurgitation severity improved to ≤2+ in all patients, with left atrial volume of (74.49±33.83) ml, left ventricular end-diastolic volume of (85.90±18.05) ml, and left ventricular end-diastolic dimension of (4.93±0.37) mm (all P<0.05). Conclusion:The short-clip strategy, focusing on precise clip placement at the commissural interface, is feasible and effective for TEER in patients with severe mitral regurgitation due to commissural prolapse.

Objectives:To quantitatively evaluate the relationship between triglyceride-glucose index and stroke risk by a dose-response meta-analysis.Methods:Prospective cohort studies on the association between triglyceride-glucose(TyG)index and stroke risk were searched by computer in China National Knowledge Infrastructure(CNKI),Wanfang Data Knowledge Service Platform,VIP,China Biomedical Literature Database,PubMed,Embase and Web of Science.The retrieval time was from the self-established database to November 7,2024.Two researchers used the Newcastle-Ottawa Scale(NOS)to evaluate literature quality and then extract relevant data for the included literatures.Stata 17.0 software was used for statistical analysis.Results:A total of 17 prospective literatures were included,involving 449 210 subjects,including 28 506 patients with stroke.The results of meta-analysis showed that the TyG index was positively correlated with the risk of stroke(HR=1.60,95%CI:1.45-1.76,P<0.05).The results of dose-response meta-analysis showed that there was a positive correlation between the TyG index and the risk of stroke,and every 1 unit increase of the TyG index,the risk of stroke increased by 20.2%.According to Egger's test,the P value is 0.962,and the P value of Begg's test is 0.967,indicating that there was no publication bias in the literature included in this study.Conclusions:There is a linear dose-response relationship between TyG index and stroke risk,and higher TyG index can increase the risk of stroke.

Objective:To investigate the clinical characteristics and prognostic significance of germline mutations in patients with myelodysplastic neoplasms (MDS) .Methods:Clinical data from 407 patients with MDS [male, 252; female, 155; median age, 64 (range, 19-85) years] diagnosed at the First Affiliated Hospital of Zhejiang University School of Medicine were retrospectively analyzed. The clinical features and prognostic effects of germline mutations were evaluated.Results:The prevalence of germline mutations in patients with MDS was 5.9% (24/407), peaking at 20.0% in the group aged 21-30 years. The spectrum of germline mutations comprised DDX41 (9 cases, 2.2%), TP53 (3 cases, 0.7%), and single cases of RUNX1, TET2, MPL, CBL, ATRX, CEBPA, ETV6, IDH1, KDM5C, SBDS, GNAS, and CTC1. Patients with germline mutations exhibited significantly lower peripheral WBC counts than those without (1.87×10 9/L vs 2.50×10 9/L, P=0.018), but showed comparable median overall survival (21.3 months vs 21.1 months, P=0.97). Patients with DDX41 germline mutations, compared with those with other germline mutations, had a significantly older median age (65 vs 54 years, P=0.010), lower WBC counts (1.51×10 9/L vs 2.31×10 9/L, P=0.040), increased mean corpuscular volume (111.80 fl vs 97.25 fl, P=0.003), and a higher prevalence of normal karyotypes (100.0% vs 53.3%, P=0.022). The most frequently co-occurring somatic mutations in DDX41 germline mutation carriers were ASXL1, TET2, and RUNX1. Conclusion:In this study, the detection rate of germline mutations in MDS patients was 5.9% (24/407), peaking at 20% in the group aged 21-30 years. DDX41 and TP53 were the most prevalent germline mutations. DDX41 mutation carriers displayed distinct clinical characteristics; however, germline mutations overall showed no significant prognostic effect.