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:Multi-facial reflective type of nano-ultrasound contrast agents（UCAs）loaded with hollow self-decomposable silica nanoparticles（SiO 2 NP）of different particle sizes were prepared to investigate the effect of the particle size of SiO 2 NP on the structural and physicochemical characteristics of the nano-UCAs，as well as on the intensity of ultrasound contrast. Methods:Two types of SiO 2 NP with large and small particle sizes（SiO 2 NP L and SiO 2 NP S）were prepared by adjusting the material ratios，and nano-UCAs with different particle sizes of SiO 2 NP S loaded at a concentration of 2 mg/ml were prepared by the thin film hydration method（SiO 2 NP L@NBs and SiO 2 NP S@NBs），and compared with the commercial SonoVue and the prepared nano-bubbles（NBs）for comparing the particle size，zeta potential and stability，scanning electron microscopy to observe the structure，CCK8 method to assess cytotoxicity，and comparison of ultrasonography imaging intensity in in vivo and in vitro experiments. Results:The particle sizes of the two SiO 2 NP（SiO 2 NP L and SiO 2 NP S）were（213.270 ± 16.890）nm and（53.870 ± 8.246）nm，respectively，and the particle size of the synthesized nano-UCA was SiO 2 NP L@NBs（628.40 ± 89.97）nm and SiO 2 NP S@NBs（493.40 ± 36.35）nm respectively，there was no difference in particle size（ P>0.05）. The scanning electron microscope showed a "fig" structure. The stability of SiO 2 NP S@NBs was better than that of SiO 2 NP L@NBs and SonoVue. Safety tests showed that both SiO 2 NP L@NBs and SiO 2 NP S@NBs were safe and usable. Compared with NBs，the intensity of contrast-enhanced ultrasound contrast in SiO 2 NP@NBs with multiple reflection interfaces was significantly enhanced，and the enhanced intensity of contrast-enhanced ultrasound in vitro and in vivo in the SiO 2 NP L@NBs group was significantly higher than that of the NBs group，the SiO 2 NP S@NBs group，and the SonoVue group（all P<0.05）. There was no significant difference in the enhanced intensity of in vitro contrast-enhanced ultrasound between the SiO 2 NP S@NBs group and that of SonoVue（ P>0.05），and the enhanced intensity of contrast-enhanced ultrasound in vivo was significantly higher than that in the SonoVue group（ P<0.05）. Conclusions:The large and small SiO 2 NPs prepared by adjusting the material ratio do not affect the nanoparticle size of UCA.The ultrasonic enhancement ability of nano-UCA with large particle size SiO 2 NP is stronger，but the stability time at room temperature is shorter than that of nano-UCA with small particle size SiO 2 NP.

Objective:With the aging of the population, the health of women's ovarian in China has received increasing attention.Ovarian aging not only affects reproductive health but also increases the risk of chronic diseases.This study aims to systematically explore the relationship between living habits and ovarian aging.Methods:Based on the data from the China Health and Retirement Longitudinal Study(CHARLS)from 2011 to 2018, 8 287 postmenopausal women were included.A retrospective cohort study was conducted using statistical analysis and machine learning methods to evaluate the impact of lifestyle factors such as smoking, drinking, sleep, and dietary patterns on the age of menopause.Results:It was found that adverse lifestyle factors such as reduced daily meal frequency [the early menopause group(2.85±0.38)times/day vs.the normal group(2.89±0.38)times/day, P=0.002], smoking [the smoking rate of the early menopause group(7.75%) vs.that of the normal group(6.15%), P=0.004], and high-frequency drinking [the early menopause rate of people who drink daily(19.76%(33/167)) vs.that of people who rarely or never drink(14.62%(1 162/7 949)), P=0.047]were associated with accelerated ovarian aging. Conclusions:Reasonable adjustment of living habits maybe help to delay the age of menopause and reduce related health risks.This study provides a scientific basis for optimizing women's healthy lifestyles, which is conducive to improving the quality of life and happiness of middle-aged and elderly women in China.

Objective:With the aging of the population, the health of women's ovarian in China has received increasing attention.Ovarian aging not only affects reproductive health but also increases the risk of chronic diseases.This study aims to systematically explore the relationship between living habits and ovarian aging.Methods:Based on the data from the China Health and Retirement Longitudinal Study(CHARLS)from 2011 to 2018, 8 287 postmenopausal women were included.A retrospective cohort study was conducted using statistical analysis and machine learning methods to evaluate the impact of lifestyle factors such as smoking, drinking, sleep, and dietary patterns on the age of menopause.Results:It was found that adverse lifestyle factors such as reduced daily meal frequency [the early menopause group(2.85±0.38)times/day vs.the normal group(2.89±0.38)times/day, P=0.002], smoking [the smoking rate of the early menopause group(7.75%) vs.that of the normal group(6.15%), P=0.004], and high-frequency drinking [the early menopause rate of people who drink daily(19.76%(33/167)) vs.that of people who rarely or never drink(14.62%(1 162/7 949)), P=0.047]were associated with accelerated ovarian aging. Conclusions:Reasonable adjustment of living habits maybe help to delay the age of menopause and reduce related health risks.This study provides a scientific basis for optimizing women's healthy lifestyles, which is conducive to improving the quality of life and happiness of middle-aged and elderly women in China.

Objective:Multi-facial reflective type of nano-ultrasound contrast agents（UCAs）loaded with hollow self-decomposable silica nanoparticles（SiO 2 NP）of different particle sizes were prepared to investigate the effect of the particle size of SiO 2 NP on the structural and physicochemical characteristics of the nano-UCAs，as well as on the intensity of ultrasound contrast. Methods:Two types of SiO 2 NP with large and small particle sizes（SiO 2 NP L and SiO 2 NP S）were prepared by adjusting the material ratios，and nano-UCAs with different particle sizes of SiO 2 NP S loaded at a concentration of 2 mg/ml were prepared by the thin film hydration method（SiO 2 NP L@NBs and SiO 2 NP S@NBs），and compared with the commercial SonoVue and the prepared nano-bubbles（NBs）for comparing the particle size，zeta potential and stability，scanning electron microscopy to observe the structure，CCK8 method to assess cytotoxicity，and comparison of ultrasonography imaging intensity in in vivo and in vitro experiments. Results:The particle sizes of the two SiO 2 NP（SiO 2 NP L and SiO 2 NP S）were（213.270 ± 16.890）nm and（53.870 ± 8.246）nm，respectively，and the particle size of the synthesized nano-UCA was SiO 2 NP L@NBs（628.40 ± 89.97）nm and SiO 2 NP S@NBs（493.40 ± 36.35）nm respectively，there was no difference in particle size（ P>0.05）. The scanning electron microscope showed a "fig" structure. The stability of SiO 2 NP S@NBs was better than that of SiO 2 NP L@NBs and SonoVue. Safety tests showed that both SiO 2 NP L@NBs and SiO 2 NP S@NBs were safe and usable. Compared with NBs，the intensity of contrast-enhanced ultrasound contrast in SiO 2 NP@NBs with multiple reflection interfaces was significantly enhanced，and the enhanced intensity of contrast-enhanced ultrasound in vitro and in vivo in the SiO 2 NP L@NBs group was significantly higher than that of the NBs group，the SiO 2 NP S@NBs group，and the SonoVue group（all P<0.05）. There was no significant difference in the enhanced intensity of in vitro contrast-enhanced ultrasound between the SiO 2 NP S@NBs group and that of SonoVue（ P>0.05），and the enhanced intensity of contrast-enhanced ultrasound in vivo was significantly higher than that in the SonoVue group（ P<0.05）. Conclusions:The large and small SiO 2 NPs prepared by adjusting the material ratio do not affect the nanoparticle size of UCA.The ultrasonic enhancement ability of nano-UCA with large particle size SiO 2 NP is stronger，but the stability time at room temperature is shorter than that of nano-UCA with small particle size SiO 2 NP.

ObjectiveTo investigate the role and possible mechanism of action of rhubarb decoction （RD） retention enema in improving inflammatory damage of brain tissue in a rat model of mild hepatic encephalopathy （MHE）. MethodsA total of 60 male Sprague-Dawley rats were divided into blank group （CON group with 6 rats） and chronic liver cirrhosis modeling group with 54 rats using the complete randomization method. After 12 weeks， 40 rats with successful modeling which were confirmed to meet the requirements for MHE model by the Morris water maze test were randomly divided into model group （MOD group）， lactulose group （LT group）， low-dose RD group （RD1 group）， middle-dose RD group （RD2 group）， and high-dose RD group （RD3 group）， with 8 rats in each group. The rats in the CON group and the MOD group were given retention enema with 2 mL of normal saline once a day； the rats in the LT group were given retention enema with 2 mL of lactulose at a dose of 22.5% once a day； the rats in the RD1， RD2， and RD3 groups were given retention enema with 2 mL RD at a dose of 2.5， 5.0， and 7.5 g/kg， respectively， once a day. After 10 days of treatment， the Morris water maze test was performed to analyze the spatial learning and memory abilities of rats. The rats were analyzed from the following aspects： behavioral status； the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） and the level of blood ammonia； pathological changes of liver tissue and brain tissue； the mRNA and protein expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （AKT）， and mammalian target of rapamycin （mTOR） in brain tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the MOD group， the RD1， RD2， and RD3 groups had a significantly shorter escape latency （all P<0.01）， significant reductions in the levels of ALT， AST， IL-1β， IL-6， TNF-α， and blood ammonia （all P<0.05）， significant alleviation of the degeneration， necrosis， and inflammation of hepatocytes and brain cells， and significant reductions in the mRNA and protein expression levels of PI3K， AKT， and mTOR in brain tissue （all P<0.05）， and the RD3 group had a better treatment outcome than the RD1 and RD2 groups. ConclusionRetention enema with RD can improve cognitive function and inflammatory damage of brain tissue in MHE rats， possibly by regulating the PI3K/AKT/mTOR signaling pathway.

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.

Objective:To explore the clinical and genetic characteristics of four children with Kabuki syndrome (KS) due to variants of KMT2D gene. Methods:Four children with KS diagnosed at the Children′s Hospital of Shanxi Province between January 2020 and December 2022 were selected as the study subjects. Whole exome sequencing was carried out for the children and their family members. Candidate variants were verified by Sanger sequencing and pathogenicity analysis.Results:The KS phenotype scores for the four children were 7, 8, 6, and 6, respectively. Child 2 also presented with a rare solitary kidney malformation. Genetic testing revealed that all children had harbored novel de novo variants of the KMT2D gene, including c. 16472_16473del, c. 858dup, c. 11899C>T, and c. 12844C>T, respectively. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), all of the variants were classified as pathogenic. Conclusion:For children showing phenotypes such as distinctive facial features, intellectual disability, developmental delay, cardiac abnormalities, and urinary system anomalies, KS should be considered. Early diagnosis and intervention can be achieved through genetic testing, especially in the presence of KMT2D gene mutations.

Objective To investigate the role of PSMA4 in the prognosis,diagnosis and immune infiltration of lung adenocarcinoma(LUAD),and explore its underlying mechanism.Methods The expression profiles and clinical data of LUAD patients were sourced from the Cancer Genome Atlas(TCGA)database.The expression level of PSMA4 in LUAD tissues(n=539)and normal tissues(n=59)were compared using the Wilcoxon rank-sum test.The expression levels of PSMA4 in LUAD tissues and normal tissues were validated by analyzing the GSE40791 and GSE10072 LUAD datasets obtained from the Gene Expression Omnibus(GEO)database.In addition,tumor and adjacent non-cancerous tissues were collected from 10 LU AD undergoing lung biopsy by fiberoptic bronchoscopy in Second Affiliated Hospital of Army Medical University from January to December 2023.The PSMA4 expression in above samples was further verified using RT-qPCR.RT-qPCR was performed to detect the expression of PSMA4 in lung cancer cells and normal lung epithelial cells.Functional enrichment analysis and immune cell infiltration analysis were conducted on the cells with high and low expression of PSMA4.Cox regression analysis and Kaplan-Meier(KM)survival analysis were used to determine the diagnostic and prognostic value of PSMA4 for LUAD,and a nomogram was constructed to predict the overall survival rate at different time points.Results The analysis of TCGA datasets,GSE40791,and GSE10072 LUAD data revealed that PSMA4 expression was significantly higher in LUAD tissues than in normal tissues(P＜0.01).RT-qPCR further confirmed that the expression of PSMA4 was obviously elevated in LUAD tissues and lung cancer cells than adjacent non-cancerous tissues and normal lung epithelial cells(P＜0.01).High PSMA4 expression could be regarded as a marker for LUAD diagnosis and poor prognosis,and was associated with reduced proportions of Tem cells,TFH cells,B cells,NK cells,Tcm cells,and mast cells in the tumor microenvironment.Conclusion PSMA4 presents significant diagnostic performance for LUAD,and is closely associated with the prognosis and immune infiltration of this malignancy.