Objective To investigate the effect of activating the N-type acetylcholine receptor(nAChR)on interleukin-18(IL-18)and PD-1 in mice with acute respiratory distress syndrome(ARDS).Methods Sixty healthy male BALB/c mice(6 weeks of age)were divided into six groups:normal(N),normal saline control(NS),normal saline+bilateral vagectomy(NS+D),ARDS+segmentation of the vagus nerve on both sides of the neck(A+D),ARDS(A),ARDS+vagal amputation,and administration of an acetylcholine receptor agonist(A+J)groups.Each group included ten mice that were fed and housed under normal conditions.Structural changes in the right lower lung were observed using fluorescence microscopy;phosphorylated nuclear factor-KB protein 65(p-NF-κBP65)levels were assessed using Western blotting;serum IL-18 and PD-1 levels were assessed using enzyme-linked immunosorbent assay(ELISA)and the double antibody sandwich method;and the percentages of CD3+and CD25+Foxp3+T lymphocytes in the middle lobe of right lung were determined using flow cytometry.Results No inflammatory cell infiltration was observed in groups N and NS.The interstitial lobes in groups A and A+D showed severe inflammatory infiltration,thickening of the alveolar wall,destruction of the alveolar structure,and loss of the alveolar cavity.Serum IL-18 and PD-1 levels in groups A and A+D were significantly higher than those in the other four groups(P＜0.05).p-NF-κBP65 and PD-1 levels in groups A and A+D were significantly higher than those in groups N,NS,and A+J(P＜0.05).CD3+and CD25+Foxp3+T cells in groups A and A+D were significantly higher than those in the other four groups(P＜0.05).Conclusion Active nAChR can inhibit IL-18 and p-NF-κBP65 through the negative regulation of T lymphocytes,decrease PD-1 expression in lung tissues,and alleviate the pathological changes of ARDS.

Objective To investigate the effect of activating the N-type acetylcholine receptor(nAChR)on interleukin-18(IL-18)and PD-1 in mice with acute respiratory distress syndrome(ARDS).Methods Sixty healthy male BALB/c mice(6 weeks of age)were divided into six groups:normal(N),normal saline control(NS),normal saline+bilateral vagectomy(NS+D),ARDS+segmentation of the vagus nerve on both sides of the neck(A+D),ARDS(A),ARDS+vagal amputation,and administration of an acetylcholine receptor agonist(A+J)groups.Each group included ten mice that were fed and housed under normal conditions.Structural changes in the right lower lung were observed using fluorescence microscopy;phosphorylated nuclear factor-KB protein 65(p-NF-κBP65)levels were assessed using Western blotting;serum IL-18 and PD-1 levels were assessed using enzyme-linked immunosorbent assay(ELISA)and the double antibody sandwich method;and the percentages of CD3+and CD25+Foxp3+T lymphocytes in the middle lobe of right lung were determined using flow cytometry.Results No inflammatory cell infiltration was observed in groups N and NS.The interstitial lobes in groups A and A+D showed severe inflammatory infiltration,thickening of the alveolar wall,destruction of the alveolar structure,and loss of the alveolar cavity.Serum IL-18 and PD-1 levels in groups A and A+D were significantly higher than those in the other four groups(P＜0.05).p-NF-κBP65 and PD-1 levels in groups A and A+D were significantly higher than those in groups N,NS,and A+J(P＜0.05).CD3+and CD25+Foxp3+T cells in groups A and A+D were significantly higher than those in the other four groups(P＜0.05).Conclusion Active nAChR can inhibit IL-18 and p-NF-κBP65 through the negative regulation of T lymphocytes,decrease PD-1 expression in lung tissues,and alleviate the pathological changes of ARDS.

Objective:To evaluate the clinical value of an obstetric intelligent assistant in predicting postpartum hemorrhage (PPH) after vaginal delivery.Methods:This retrospective cohort study included 4 832 women who delivered vaginally at ≥26 weeks of gestation at the Third Affiliated Hospital, Guangzhou Medical University between May 2023 and April 2025. Participants were categorized into PPH (382 cases, blood loss ≥500 ml within 24 h after delivery) and non-PPH groups (4 450 cases). Using traditional statistical methods combined with machine learning approaches, including support vector machines and extreme gradient boosting, supplemented with deep learning techniques, we developed a novel artificial neural network model—the obstetric intelligent assistant. This model provides a refined classification of PPH occurrence and estimated blood loss volume. The obstetric intelligent assistant integrates 70 antenatal and intrapartum risk factors through hospital information system interfacing to generate visualized risk probability outputs. Predictive performance was compared between the obstetric intelligent assistant and four conventional prediction tools (Chinese Labor Room Traffic Light System; Association of Women's Health, Obstetric and Neonatal Nurses; American College of Obstetrics and Gynecology Safe Motherhood Initiative; and California Maternal Quality Care Collaborative prediction tools) using receiver operating characteristic curve.Results:(1) For antenatal prediction, the obstetric intelligent assistant achieved an area under the curve of 0.826 (95% CI: 0.774-0.838), with sensitivity of 0.794 and specificity of 0.712, while the four conventional prediction tools showed area under the curve ranging from 0.569 to 0.586. (2) For intrapartum prediction, the obstetric intelligent assistant achieved an area under the curve of 0.786 (95% CI: 0.751-0.820), with sensitivity of 0.837 and specificity of 0.762, whereas the conventional tools showed area under the curve between 0.600 and 0.613. Conclusion:The obstetric intelligent assistant demonstrates superior performance in predicting PPH compared to conventional prediction tools.

Objective:To evaluate the clinical value of an obstetric intelligent assistant in predicting postpartum hemorrhage (PPH) after vaginal delivery.Methods:This retrospective cohort study included 4 832 women who delivered vaginally at ≥26 weeks of gestation at the Third Affiliated Hospital, Guangzhou Medical University between May 2023 and April 2025. Participants were categorized into PPH (382 cases, blood loss ≥500 ml within 24 h after delivery) and non-PPH groups (4 450 cases). Using traditional statistical methods combined with machine learning approaches, including support vector machines and extreme gradient boosting, supplemented with deep learning techniques, we developed a novel artificial neural network model—the obstetric intelligent assistant. This model provides a refined classification of PPH occurrence and estimated blood loss volume. The obstetric intelligent assistant integrates 70 antenatal and intrapartum risk factors through hospital information system interfacing to generate visualized risk probability outputs. Predictive performance was compared between the obstetric intelligent assistant and four conventional prediction tools (Chinese Labor Room Traffic Light System; Association of Women's Health, Obstetric and Neonatal Nurses; American College of Obstetrics and Gynecology Safe Motherhood Initiative; and California Maternal Quality Care Collaborative prediction tools) using receiver operating characteristic curve.Results:(1) For antenatal prediction, the obstetric intelligent assistant achieved an area under the curve of 0.826 (95% CI: 0.774-0.838), with sensitivity of 0.794 and specificity of 0.712, while the four conventional prediction tools showed area under the curve ranging from 0.569 to 0.586. (2) For intrapartum prediction, the obstetric intelligent assistant achieved an area under the curve of 0.786 (95% CI: 0.751-0.820), with sensitivity of 0.837 and specificity of 0.762, whereas the conventional tools showed area under the curve between 0.600 and 0.613. Conclusion:The obstetric intelligent assistant demonstrates superior performance in predicting PPH compared to conventional prediction tools.

Objective To investigate the effects of α7 nicotinic acetylcholine receptor (α7nAChR) on CD11b, IL-1β, IL-18 and TNF-α in acute respiratory distress syndrome (ARDS) mice. Methods A total of 40 healthy and clean male Balb/C mice (6 weeks old) were randomly divided into normal group (N group), normal saline control group (NS group), ARDS group (A group), and ARDS mice treated with nicotinic acetylcholine receptor agonist after bilateral cervical vagotomy group (J group), with 10 mice in each group. The right lung structure of mice in each group was observed by hematoxylin-eosin (HE) staining, the lung tissue wet weight/body weight ratio (LWW/DW ratio) was detected, and the percentage of CD11b in the alveolar lavage fluid of mice was detected by flow cytometry. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of IL-1β mRNA, IL-18 mRNA and TNF-α mRNA in left lung tissue. Serum IL-18 was determined by enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich method. Results HE staining of the right lung of mice in group N and NS showed normal structure, while the lung interstitial of mice in group A showed a large number of inflammatory cells infiltrated, alveolar wall thickened, alveolar structure destroyed and alveolar cavity fused. The alveolar structure of mice in group J was intact, with a little damage and alveolar cavity. The percentage of CD11b in alveolar lavage fluid in group A was higher than that in the other three groups, and the difference was statistically significant compared with group N, NS and J, respectively (P<0.05). The expressions of IL-1β mRNA, IL-18 mRNA and TNF-α mRNA in the left lung of mice in group J were statistically significant compared with those in group N, NS and A (P<0.05), and the serum IL-18 level of mice in group A was higher than that in the other three groups, and the differences were statistically significant compared with groups N, NS and J, respectively (P<0.05). Conclusions Activation of α7nAChR can directly inhibit the release of CD11b in lung tissue and reduce the accumulation of inflammatory factors. Simultaneously, it can also directly inhibit the expression of IL-β1 mRNA, IL-18 mRNA and TNF-α mRNA in lung tissue and the release of IL-18, thus inhibiting the inflammatory response of ARDS and alleviating the pathological changes of ARDS.

Objectives To explore the expression, biological function, and mechanism of MKI67 in pancreatic cancer and its clinical significance. Methods The expression level, diagnosis, and prognostic value of MKI67 in pancreatic cancer were analyzed using public databases. We also investigated the association between the MKI67 with immune cell infiltration and immune checkpoint molecules. We analyzed the functional pathway enrichment to uncover the possible molecular mechanisms. qRT-PCR and Western blot assay were used to verify the expression of MKI67 mRNA and protein. Immunohistochemistry staining was used to detect the expression of MKI67 in tissue protein. Results The high expression of MKI67 was significantly associated with high histological grades and poor outcomes in pancreatic cancer. High MKI67 expression was correlated with poor prognosis of pancreatic cancer patients (P=0.009). MKI67 was an independent risk factor for the patient outcome (95%CI: 1.084-1.743, P<0.05). The MKI67 expression was positively correlated with the helper T cell 2 levels but negatively correlated with plasmacytoid DC, NK cells, mast cells, the T follicular helper, immune DC, and CD8 T cells. Conclusion MKI67 may serve as a biomarker for the diagnosis and prognosis of pancreatic cancer and the mechanism might be associated with immune escape or immunosuppression.

Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

Objective By investigating the effects of miR-379-5p on the proliferation,invasion and metastasis of mouse breast cancer 4T1 cells,we aimed to provide new therapeutic targets for the clinical inhibition of breast cancer proliferation,invasion,and metastasis.Methods After plasmid transfection,4T1 cells were utilized to detect the expression of miR-379-5p using fluorescence quantitative PCR.While 5-ethynyl-2'doxyuridine(EdU)cell proliferation and Transwell assays were employed to detect changes in the proliferation and invasion ability of 4T1 cells in each group.The migration ability of 4T1 cells after overexpression and knockdown of miR-379-5p was examined by scratch healing assay.A transplanted tumor model of breast cancer was established in BABL/c mice,and the effects of overexpressing miR-379-5p on tumor growth and the number and size of lung metastases were observed.Results EdU result showed that knocking down miR-379-5p enhanced the proliferation ability of the cells compared with the control group cells,and miR-379-5p overexpression reduced the capacity of breast cancer cells to proliferate(P＜0.05).Transwell and wound healing assays showed that miR-379-5p knockdown enhanced,while miR-379-5p overexpression significantly inhibited,the invasion and migratory ability of breast cancer cells(P＜0.01).An in vivo tumorigenesis experiment with BABL/c mice showed that miR-379-5p overexpression significantly slowed the tumor growth rate(P＜0.05)and inhibited lung metastasis(P＜0.01).Conclusions MiR-379-5p plays a role in tumor gene suppression in breast cancer and inhibits the proliferation,invasion,and migration of mouse breast cancer 4T1 cells.

Objective:To analyze the relationship between dietary composition of residents in endemic fluorosis areas and skeletal fluorosis.Methods:A case-control study was used to analyze the difference of dietary composition between patients with skeletal fluorosis (case group) and residents without skeletal fluorosis (control group). In August 2019, taking the drinking water-borne endemic fluorosis area in Wenshui County, Lvliang City, Shanxi Province as the survey site, a cluster sampling method was adopted to select local residents aged over 18 years old, and a questionnaire survey was conducted by face-to-face interview. The survey contents included gender, age and consumption frequency of various foods. Binary logistic regression was used to analyze the relationship between food consumption frequency and skeletal fluorosis. The diagnosis of skeletal fluorosis was made by using portable digital radiography (DR) to take X-ray films of forearm and lower leg, combining with clinical signs, and according to the Diagnostic Standard for Endemic Skeletal Fluorosis (WS/T 192-2008) to determine.Results:A total of 1 061 subjects were included in this study, including 376 in the case group and 685 in the control group. The age composition of patients in the case group (≤60, > 60 years old: 162, 214 cases) was significantly different from that in the control group (≤60, > 60 years old: 423, 261 cases, χ 2 = 34.52, P < 0.001). There was no statistically significant difference in gender ratio (χ 2 = 1.37, P = 0.251). The proportion of patients in the case group who ate meat and eggs > 1 time/week was lower than that in the control group (χ 2 = 8.06, 5.46, P < 0.05), the proportion of patients who ate milk > 1 time/week was higher than that in the control group (χ 2 = 4.01, P = 0.046), and the proportion of patients who ate seafood ≥1 time/week was lower than that in the control group (χ 2 = 4.16, P = 0.046). The results of binary logistic regression analysis showed that after adjusting for age, sex, and urinary fluoride, the frequency of eating meat, eggs or milk > 1 time/week and the frequency of eating seafood ≥1 time/week were not related to the risk of skeletal fluorosis ( P > 0.05); however, in the group ≤60 years old, the frequency of eating eggs > 1 time/week was associated with the risk of skeletal fluorosis [odds ratio ( OR) = 0.59, 95% confidence interval ( CI): 0.39, 0.88]. Conclusions:The consumption frequency of meat, milk, eggs and seafood is significantly different between the skeletal fluorosis patients and the control people. In the population ≤60 years old, consumption frequency of eggs > 1 time/week may reduce the risk of skeletal fluorosis.

Non-alcoholic fatty liver disease (NAFLD) is a series of chronic liver diseases strongly associated with the metabolic disorder with an increasing rate of worldwide prevalence.Due to its complicated pathogenesis, only Saroglitazar has been approved by Indian Drug Controller General (DCGI) as a PPAR-α/γ dual agonist to treat non-cirrhotic non-alcoholic steatohepatitis.Combination therapy, which can target same or different signaling pathways of NAFLD pathogenesis, has been developed to achieve synergistic therapeutic efficacy.Currently, small-molecule drug combination, RNAi combination therapy, and chemogene therapy are proposed as promising strategies in NAFLD treatment.In addition, designing a smart, safe and effective drug delivery system is key to realizing the druggability, clinical translation and industrialization of small molecule drugs and gene drugs.This review summarizes the research status and delivery system of small-molecule drug combination, RNAi combination therapy, and chemogene therapy, in the hope of providing some novel insight for the treatment of NAFLD.