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 estimate the economic burden of 14 chronic diseases among middle-aged and elderly people in China.Methods:This cross-sectional study was based on longitudinal data from five waves (2011 to 2020) of the China Health and Retirement Longitudinal Study (CHARLS). Participants aged 45 and over who had been diagnosed with exactly one target chronic disease were included in the study. The 14 chronic diseases included: hypertension, dyslipidemia, diabetes or elevated blood glucose, malignant tumors, chronic pulmonary diseases, liver diseases, heart diseases, stroke, kidney diseases, digestive system diseases, emotional and mental diseases, memory-related diseases, arthritis or rheumatic diseases, and asthma. The economic burden of disease was measured in terms of both direct and indirect economic burden, with the results adjusted using a healthcare-specific Consumer Price Index (CPI). The direct economic burden included direct medical and non-medical burden. The human capital method was employed to calculate the indirect economic burden. A generalized linear mixed model (GLMM) was conducted to compare the differences in the economic burden between urban and rural areas, with provinces and prefecture-level cities set as random effects and residence (urban or rural areas) as fixed effects to control for the effects of geographic hierarchical structure. Gender, age and educational attainment were also included as covariates to control for confounding factors. The model′s robustness was assessed by comparing the significance of urban-rural differences before and after adding the covariates.Results:The median annual economic burden of the 14 chronic diseases among the middle-aged and elderly population in China ranged from 7 565 to 17 174 CNY, of which the direct economic burden ranged from 6 909 to 16 565 CNY, and the indirect burden ranged from 284 to 1 276 CNY. The direct economic burden was primarily driven by direct medical burden (83.67% to 95.01% of direct economic burden). Out-of-pocket expenses for outpatient medical burden ranged from 50% to 100%, while those for inpatient ranged from 36.30% to 61.29%. GLMM analysis revealed no statistically significant difference in the overall economic burden between urban and rural areas across diseases. However, the burden of inpatient medical burden for arthritis or rheumatism was higher in urban areas than in rural areas (5 338 vs. 3 898 CNY; LR=6.04, P=0.014). Similarly, the burden of self-treatment for hypertension was also higher in urban areas than in rural areas (324 vs. 238 CNY; LR=8.30, P=0.004). The outpatient non-medical burden for diabetes or elevated blood glucose (59 vs. 149 CNY; LR=5.99, P=0.014), stroke (0 vs. 307 CNY; LR=4.55, P=0.033), and digestive system diseases (45 vs. 107 CNY; LR=9.58, P=0.002) was lower in urban areas than rural. Conclusions:Chronic diseases cause heavy economic burden on middle-aged and elderly people in both urban and rural areas of China, with direct economic burden accounting for the majority of expenditure. The outpatient medical burden accounts for a higher proportion of out-of-pocket expenses than the inpatient.

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 investigate the prognosis of patients with triple-negative invasive lobular carcinoma (TN-ILC) undergoing breast-conserving surgery combined with radiotherapy (BCS+RT) versus total mastectomy. Methods A retrospective analysis was performed for 2 386 female patients with TN-ILC who underwent surgery in the SEER database from 2006 to 2018, and the baseline characteristics (age, histological grade, AJCC stage, etc.) were balanced by propensity score matching (PSM, 1∶1, caliper value 0.02), and breast cancer-specific survival (BCSS) and overall survival (OS) were compared by Kaplan-Meier method and COX regression analysis. Results A total of 1 056 pairs of patients were obtained after PSM, and the BCS+RT group had significantly better BCSS and OS than the total mastectomy group (both P<0.001). Stratified analyses showed that BCS+RT had a survival advantage in all subgroups except histologic grade Ⅰ and tumor stage Ⅰ. Multivariate analysis confirmed that BCS+RT was an independent protective factor (BCSS: HR=0.682, OS: HR=0.607, both P<0.001). Conclusions BCS+RT significantly improves survival compared with total mastectomy in patients with TN-ILC, supporting BCS+RT as the preferred treatment strategy for eligible patients.

Objective: To investigate the role of STAT3 phosphorylation in ferroptosis regulation and its impact on cisplatin resistance mechanisms in human osteosarcoma cells． Methods: Human osteosarcoma HOS cells and cispl- atin-resistant HOS / DDP cells were treated with cisplatin ( 0. 5，1，2，4，8，16，32 mg / L) ，the ferroptosis inducer Erastin ( Era) ( 2 μmol / L) ，and / or the ferroptosis inhibitor Ferrostatin-1 ( Fer1) ( 10 μmol / L) ．Cell viability and proliferation were assessed using the Cell Counting Kit-8 ( CCK-8) and colony formation assays，and cell migration was evaluated via a scratch assay．Reactive oxygen species ( ROS) ，intracellular ferrous iron levels，mitochondrial membrane potential，mitochondrial function，malondialdehyde ( MDA) levels，and the reduced glutathione / oxi- dized glutathione ( GSH / GSSG) ratio were measured using commercial kits．The mRNA expression of ferroptosis- related genes was analyzed by quantitative reverse transcription polymerase chain reaction ( RT-qPCR) ．The protein levels of glutathione peroxidase 4 ( GPX4) ，solute carrier family 7 member 11 ( SLC7A11) ，phosphorylated STAT3 ( p-STAT3) ，and total STAT3 were determined by Western blot． Results: With cisplatin treatment，HOS cells ex- hibited decreased cell viability，mitochondrial membrane potential，and GSH / GSSG ratio ( P＜0. 01) ，along with elevated levels of ROS，ferrous ion，and MDA content ( P ＜0. 01) ． The protein levels of GPX4 ( P ＜0. 01) ， SLC7A11，and p-STAT3 also decreased ( P ＜0. 05) ．Coadministration with the ferroptosis inhibitor Ferrostatin-1 ( Fer-1) reversed these aforementioned effects ( P ＜0. 05) ．In HOS / DDP cells，the mRNA levels of ferroptosis- suppressive genes ( GPX4，FTH1，SLC7A11，and AIFM2) were significantly higher than those in HOS cells ( P < 0. 05) ，whereas the expression of ferroptosis-promoting genes ( ACSL4 and PTGS2) was significantly lower ( P < 0. 05) ．The cisplatin-induced reductions in cell viability and mitochondrial membrane potential，as well as the in- creases in ROS，ferrous ion，and MDA levels，were less pronounced in HOS / DDP cells than in HOS cells．The SLC7A11 protein level showed no significant change． However，combined treatment with the ferroptosis inducer Erastin ( Era) resulted in significant decreases in viability，mitochondrial membrane potential，and the GSH / GSSG ratio in HOS / DDP cells ( P＜0. 05) ．Furthermore，the protein levels of p-STAT3，GPX4，and SLC7A11 were also markedly reduced ( P＜0. 05) ． Conclusion The activation of ferroptosis mediated by p-STAT3 enhances cisplatin sensitivity in HOS / DDP cells．

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 evaluate the clinical efficacy of modified Yulinzhu Formula combined with plum-blossom needle tapping acupuncture in treating kidney-yang deficiency type of ovulatory dysfunction infertility(ODI).Methods A total of 100 ODI patients diagnosed at Hengshui Hospital of Traditional Chinese Medicine between January 2023 and January 2024 were enrolled and randomly divided into an observation group and a control group using a random number table,with 50 cases in each group.The control group received conventional western medical treatment,while the observation group received additional oral administration of modified Yulinzhu Formula and plum-blossom needle tapping acupuncture therapy.Treatment was administered for one menstrual cycle as a course,with three consecutive courses completed.After three months,clinical efficacy was assessed by comparing traditional Chinese medicine(TCM)syndrome scores,ovulation rate,pregnancy rate,proportion of type A endometrium,ovarian volume,antral follicle count(AFC),and adverse reactions between groups.Results(1)After treatment,the scores of TCM syndromes in the two groups were significantly improved(P＜0.05),and the improvement in the observation group was superior to that in the control group,the difference being significant(P＜0.05).(2)The total effective rate was 90.00％(45/50)in the observation group and 72.00％(36/50)in the control group.The curative effect of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).(3)The ovulation rate,pregnancy rate and the proportion of type A endometrium in the observation group were significantly superior to those in the control group,and the differences were statistically significant(P＜0.05).(4)After treatment,the ovarian volume and the number of antral follicles in the two groups were significantly improved(P＜0.05),and the improvement in the ovarian volume and the number of antral follicles in the observation group was superior to that in the control group,the difference being significant(P＜0.05).(5)The incidence of adverse reactions was 8.00％(4/50)in the observation group and 24.00％(12/50)in the control group.The incidence of adverse reactions in the observation group was significantly lower than that in the control group,the difference was statistically significant(P＜0.05).Conclusion Modified Yulinzhu Formula combined with plum-blossom needle tapping acupuncture effectively improves clinical symptoms,enhances ovulation/pregnancy rates,increases type A endometrium,optimizes ovarian volume and AFC,and reduces adverse reactions in kidney-yang deficiency ODI patients.