Objectives To explore the causal relationship between asthma-related pneumonia and heart failure(HF)by using two-sample Mendelian randomization(MR)analysis.Methods The causal relationship between asthma-related pneumonia and HF were systematically evaluated through two-sample MR analysis based on publicly available Genome-Wide Association Study data.Inverse variance weighted(IVW),MR-Egger,weighted median,weighted mode,and simple mode methods were used.Different models were used to analyze sensitivity.Testing approaches were conducted to assess the robustness of the results.Results IVW analysis results showed that there was a causal relationship between asthma-related pneumonia and HF risk(OR=1.113,95％CI:1.027-1.206,P=0.009).HF was not found to have a causal effect on asthma-related pneumonia risk based on IVW analysis(OR=1.018,95％CI:0.780-1.328,P=0.893).The variance used in all MR analyses showed no heterogeneity or horizontal pleiotropy.The leave-one-out analysis revealed that no single nucleotide polymorphism disproportionately influenced the results,further confirming the credibility of the bidirectional MR findings.Conclusion This study established a causal relationship between asthma-related pneumonia and HF risk.No clear causal relationship between HF and asthma-related pneumonia risk was found.

Objective:To explore the regulation and mechanism of human placental extract(HPE)on lipopolysaccharide(LPS)-induced BV2 microglial cell inflammation.Methods:Microglia cell lines(BV2)were cultured in vitro and divided into PBS group,HPE group,LPS group and LPS+HPE group.BV2 cell viability was measured by CCK-8 analysis.A fluorescent probe targeting reactive oxygen species(ROS)was to detect the level of intracellular ROS.The mRNA levels of TNF-α,IL-1β and IL-6 were detected by RT-qPCR.The supernatants of different treatment groups were collected.The content of nitric oxide(NO)was detected by the Griess method,and the secretion levels of TNF-α,IL-1β and IL-6 were detected by the flow magnetic bead microarray(CBA)method.The indirect contact co-culture system between BV2 and mouse hippocampal neurons cell line HT22 cells was established to evaluate the neurotoxicity of HPE by the assessment of the cell viability and apoptosis of HT22 cells using CCK-8 or flow cytometry.The potential signaling molecules of NF-κB signaling pathway was detected by flow cytometry.Results:Compared with the PBS group,1.2 μg/ml LPS and 50 ng/ml HPE significantly inhibited the activity of BV2 cells.Compared with the PBS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells of the LPS group were significantly increased(P<0.05),and the secretion levels of NO,TNF-α,IL-1βand IL-6 in the supernatant were also significantly upregulated(P<0.05).The expressions of related signaling pathway molecules Toll-like receptor 4(TLR4),pIκBα and pNF-κB p65 were significantly increased(P<0.05).Compared with the LPS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells,the secretion levels of NO,TNF-α,IL-1β and IL-6,the neurotoxicity and neuronal apoptosis induced by microglial conditional medium,and the expressions of TLR4,pIκBα and pNF-κB p65 in the LPS+HPE group were significantly downregulate(P<0.05).Conclusion:HPE may alleviate the microglial inflammation,possibly through the TLR4/NF-κB signaling pathway.

Objective:To explore the clinical significance of skeletonized lymph node dissection of No.12 lymph nodes after neoadjuvant therapy in patients with advanced gastric cancer.Methods:For this retrospective case-cohort study we collected data from patients with advanced gastric cancer who underwent neoadjuvant chemotherapy and D2 or more extensive curative resection including No.12 lymph node dissection at the First Affiliated Hospital of Sun Yat-sen University from January, 2011 to December, 2022. Patients were divided into two groups based on whether they received skeletonized dissection of No.12 lymph nodes: 177 cases were in the skeletonized group, and 55 cases were in the nonskeletonized group. The differences of prognosis between the two groups were compared, and logistic regression models were used to analyze the factors affecting No.12 lymph node metastasis in the overall cohort and No.12b or No.12p lymph node metastasis in the skeletonized group.Results:A total of 232 patients were included, with 84 females (36.2%) and 148 males (63.8%), with an average age of 56.4±11.6 years. The proportion of female and ycT4 patients was significantly higher in the skeletonized group than in the nonskeletonized group (both P<0.05). Among all 232 patients, No. 12a metastasis occurred in 14 cases (6.0%). In the skeletonized group of 177 patients, No. 12b and No. 12p metastases were observed in 6 patients each (3.4%), and 4 patients had concurrent metastases in both No. 12b and No. 12a. The 5-year overall survival (OS) rates were 45.5% in the skeletonized group and 42.8% in the nonskeletonized group, with no statistical difference (HR=0.755, 95%CI: 0.488-1.168, P=0.580). The 5-year disease-free survival (DFS) rates were 39.8% and 41.0%, respectively, also with no statistical difference (HR=0.775, 95%CI: 0.513-1.172, P=0.584). 5-year OS for patients without No.12 lymph node metastasis was 48.8%, which was higher than the 15.9% for those with metastasis (HR=0.349, 95% CI: 0.209-0.584, P=0.003). Additionally, the 5-year DFS for those without metastasis was 44.3%, significantly higher than the 5.7% for those with metastasis (HR=0.444, 95%CI: 0.276-0.716, P<0.001). For patients without No. 12b or No. 12p lymph node metastasis, the 5-year OS was 47.6%, and the 5-year DFS was 42.3%, both of which were significantly higher than the 16.7% and 8.3% for those with No.12b or No. 12p lymph node metastasis, respectively (HR=0.353, 95%CI: 0.183-0.681, P=0.005; HR=0.457, 95%CI: 0.244-0.855, P=0.006). Multivariate analysis showed that more advanced ypN stage (OR=3.908, 95%CI:1.638-9.323, P=0.002) and tumor location in the lower stomach or whole stomach (OR=3.533, 95%CI: 1.312-9.511, P=0.012) were independent risk factors for No.12 lymph node metastasis and also for No.12b and No.12p lymph node metastasis (OR=2.426, 95%CI: 1.212-4.856, P=0.012 and OR=4.908, 95%CI:1.182-20.373, P=0.028, respectively). Conclusion:Patients with advanced gastric cancer who have more advanced ypN stage and tumor location in the lower stomach or whole stomach have a higher risk of No.12b and No.12p metastasis and thus require further skeletonized lymph node dissection of No.12.

Objective:To explore the regulation and mechanism of human placental extract(HPE)on lipopolysaccharide(LPS)-induced BV2 microglial cell inflammation.Methods:Microglia cell lines(BV2)were cultured in vitro and divided into PBS group,HPE group,LPS group and LPS+HPE group.BV2 cell viability was measured by CCK-8 analysis.A fluorescent probe targeting reactive oxygen species(ROS)was to detect the level of intracellular ROS.The mRNA levels of TNF-α,IL-1β and IL-6 were detected by RT-qPCR.The supernatants of different treatment groups were collected.The content of nitric oxide(NO)was detected by the Griess method,and the secretion levels of TNF-α,IL-1β and IL-6 were detected by the flow magnetic bead microarray(CBA)method.The indirect contact co-culture system between BV2 and mouse hippocampal neurons cell line HT22 cells was established to evaluate the neurotoxicity of HPE by the assessment of the cell viability and apoptosis of HT22 cells using CCK-8 or flow cytometry.The potential signaling molecules of NF-κB signaling pathway was detected by flow cytometry.Results:Compared with the PBS group,1.2 μg/ml LPS and 50 ng/ml HPE significantly inhibited the activity of BV2 cells.Compared with the PBS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells of the LPS group were significantly increased(P<0.05),and the secretion levels of NO,TNF-α,IL-1βand IL-6 in the supernatant were also significantly upregulated(P<0.05).The expressions of related signaling pathway molecules Toll-like receptor 4(TLR4),pIκBα and pNF-κB p65 were significantly increased(P<0.05).Compared with the LPS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells,the secretion levels of NO,TNF-α,IL-1β and IL-6,the neurotoxicity and neuronal apoptosis induced by microglial conditional medium,and the expressions of TLR4,pIκBα and pNF-κB p65 in the LPS+HPE group were significantly downregulate(P<0.05).Conclusion:HPE may alleviate the microglial inflammation,possibly through the TLR4/NF-κB signaling pathway.

Objective:To explore the clinical significance of skeletonized lymph node dissection of No.12 lymph nodes after neoadjuvant therapy in patients with advanced gastric cancer.Methods:For this retrospective case-cohort study we collected data from patients with advanced gastric cancer who underwent neoadjuvant chemotherapy and D2 or more extensive curative resection including No.12 lymph node dissection at the First Affiliated Hospital of Sun Yat-sen University from January, 2011 to December, 2022. Patients were divided into two groups based on whether they received skeletonized dissection of No.12 lymph nodes: 177 cases were in the skeletonized group, and 55 cases were in the nonskeletonized group. The differences of prognosis between the two groups were compared, and logistic regression models were used to analyze the factors affecting No.12 lymph node metastasis in the overall cohort and No.12b or No.12p lymph node metastasis in the skeletonized group.Results:A total of 232 patients were included, with 84 females (36.2%) and 148 males (63.8%), with an average age of 56.4±11.6 years. The proportion of female and ycT4 patients was significantly higher in the skeletonized group than in the nonskeletonized group (both P<0.05). Among all 232 patients, No. 12a metastasis occurred in 14 cases (6.0%). In the skeletonized group of 177 patients, No. 12b and No. 12p metastases were observed in 6 patients each (3.4%), and 4 patients had concurrent metastases in both No. 12b and No. 12a. The 5-year overall survival (OS) rates were 45.5% in the skeletonized group and 42.8% in the nonskeletonized group, with no statistical difference (HR=0.755, 95%CI: 0.488-1.168, P=0.580). The 5-year disease-free survival (DFS) rates were 39.8% and 41.0%, respectively, also with no statistical difference (HR=0.775, 95%CI: 0.513-1.172, P=0.584). 5-year OS for patients without No.12 lymph node metastasis was 48.8%, which was higher than the 15.9% for those with metastasis (HR=0.349, 95% CI: 0.209-0.584, P=0.003). Additionally, the 5-year DFS for those without metastasis was 44.3%, significantly higher than the 5.7% for those with metastasis (HR=0.444, 95%CI: 0.276-0.716, P<0.001). For patients without No. 12b or No. 12p lymph node metastasis, the 5-year OS was 47.6%, and the 5-year DFS was 42.3%, both of which were significantly higher than the 16.7% and 8.3% for those with No.12b or No. 12p lymph node metastasis, respectively (HR=0.353, 95%CI: 0.183-0.681, P=0.005; HR=0.457, 95%CI: 0.244-0.855, P=0.006). Multivariate analysis showed that more advanced ypN stage (OR=3.908, 95%CI:1.638-9.323, P=0.002) and tumor location in the lower stomach or whole stomach (OR=3.533, 95%CI: 1.312-9.511, P=0.012) were independent risk factors for No.12 lymph node metastasis and also for No.12b and No.12p lymph node metastasis (OR=2.426, 95%CI: 1.212-4.856, P=0.012 and OR=4.908, 95%CI:1.182-20.373, P=0.028, respectively). Conclusion:Patients with advanced gastric cancer who have more advanced ypN stage and tumor location in the lower stomach or whole stomach have a higher risk of No.12b and No.12p metastasis and thus require further skeletonized lymph node dissection of No.12.

Objectives To explore the causal relationship between asthma-related pneumonia and heart failure(HF)by using two-sample Mendelian randomization(MR)analysis.Methods The causal relationship between asthma-related pneumonia and HF were systematically evaluated through two-sample MR analysis based on publicly available Genome-Wide Association Study data.Inverse variance weighted(IVW),MR-Egger,weighted median,weighted mode,and simple mode methods were used.Different models were used to analyze sensitivity.Testing approaches were conducted to assess the robustness of the results.Results IVW analysis results showed that there was a causal relationship between asthma-related pneumonia and HF risk(OR=1.113,95％CI:1.027-1.206,P=0.009).HF was not found to have a causal effect on asthma-related pneumonia risk based on IVW analysis(OR=1.018,95％CI:0.780-1.328,P=0.893).The variance used in all MR analyses showed no heterogeneity or horizontal pleiotropy.The leave-one-out analysis revealed that no single nucleotide polymorphism disproportionately influenced the results,further confirming the credibility of the bidirectional MR findings.Conclusion This study established a causal relationship between asthma-related pneumonia and HF risk.No clear causal relationship between HF and asthma-related pneumonia risk was found.

The purpose of this study was to investigate the damage mechanism of pathogenic E.coli on mouse brain microvascular endothelial cells(BMEC cells)and mouse alveolar macrophages(MH-S cells),as well as the lung and brain of healthy mice.In this study,BMEC cells and MH-S cells were infected with pathogenic E.coli strains,and cell morphological changes were observed.Plate counting method was used to detect the adhesion and invasion ability of the strains to cells and the number of bacteria in the lungs and brains of mice.RT-qPCR was used to detect the ex-pression of TNF-α,IL-1β and IL-6 genes in cells and mouse organs at different time periods.West-ern blot was used to detect the expression of p-NF-κB,p-JAK2 and p-STAT3 proteins related to inflammation in cells and mouse organs after infection.The results showed that the cell culture medium of the infection group was turbid,the cell vision became dark and blurred,some cells shrank and died,and more fragments were produced.The adhesion rate and invasion rate of BMEC cells at 3 h were significantly lower than those at 6 h(P＜0.050),and the adhesion rate and inva-sion rate of MH-S cells at 3 h were significantly higher than those at 6 h(P＜0.010).Infected mice had a large area of swelling and bleeding in the brain,and the lungs had different degrees of swell-ing and bleeding.The bacterial load in the brain and lung was the highest at 12 h.Compared with the control group,the mRNA expression levels of IL-1β,IL-6 and TNF-α in the infection group were significantly increased at 3 h and 6 h(P＜0.050),and the mRNA expression levels of inflam-matory factors in BMEC cells and MH-S cells were the highest at 6 and 3 h,respectively.The mR-NA expression of inflammatory factors in the brain and lung of infected mice showed a trend of in-creasing first and then decreasing with time,with the highest expression at 12 h after infection.The expression levels of p-NF-κB protein in BMEC cells,MH-S cells,lung and brain tissues of mice in the infection group were significantly higher than those in the control group(P＜0.001),and the expression levels of p-JAK2 protein and p-STAT3 protein were significantly lower than those in the control group(P＜0.050).The above results showed that pathogenic E.coli could adhere and invade BMEC cells and MH-S cells,colonize in lung and brain tissues of mice,promote the expres-sion of NF-κB protein in cells and tissues,inhibit the expression of JAK2 protein and STAT3 pro-tein,and then stimulate cells and tissues to produce inflammatory response.

Objective:To investigate potential mechanisms of Niclosamide,a calcium-binding protein S100A4 inhibitor,in regulating inflammatory response of bronchial epithelial cells.Methods:Human bronchial epithelial cells BEAS-2B were cultured in vitro and stimulated by LPS or Niclosamide-pretreatment.Inflammatory cytokines and potential signaling molecules expressions were determined by RT-qPCR and Western blot.Intracellular ROS level was quantified by fluorescent probe.Results:Increased ROS level was observed in LPS-stimulated and Niclosamide-pretreatment cells(P<0.05).Compared with control group,mRNA and protein expressions of S100A4 were increased(P<0.05),TLR4/STAT3,MAPK1/3/SIRT1,NF-κB/IKKβ/p65 mRNA expressions were increased(P<0.05),inflammatory cytokines IL-1β and IL-6,tight junction Occludin and ZO-1 mRNA expressions were increased after LPS-treatment(P<0.05),whereas these genetic expressions were downregulated by Niclosamide-pretreatment(P<0.05),except for TLR4/MAPK1 and NF-κB/IKKβ/p65(P>0.05).Western blot showed that compared with control group,LPS-stimulation promoted protein expressions of S100A4,STAT3,MAPK3/SIRT1,IL-1β and TNF-α(P<0.05),while downregulated protein expression of Occludin.Compared with LPS group,niclosamide-pretreatment downregulated protein expressions of S100A4,STAT3,MAPK3/SIRT1,IL-1β and TNF-α(P<0.05),while restored protein expression of Occludin(P<0.05).Conclusion:Calcium-binding protein S100A4 inhibitor Niclosamide can alleviate S100A4 expression and inflammatory response of bronchial epithelial cells,which is poten-tially related to STAT3 or MAPK3/SIRT1 signaling.

Purpose To investigate the effect of MYD88 gene overexpression on the proliferation and apoptosis of human diffuse large B cell lymphoma(DLBCL)cells,and to prelimi-narily explore the mechanism of MYD88 gene action.Methods PEGFP-C2-MYD88 overexpressing MYD88 L265P gene was transfected into DLBCL cells by plasmid transfection.The exper-iment was divided into blank control group,negative control group and MYD88 L265P overexpression group.The fluores-cence expression of MYD88 L265P after overexpression was ob-served under inverted fluorescence microscope.RT-PCR and Western blot were used to detect the mRNA and protein expres-sion of MYD88 L265P,IRAK4,NF-κB and BCL2 in DLBCL cells before and after overexpression of MYD88 L265.CCK8 method was used to detect DLBCL cells proliferation and Ho-echst staining was used to detect DLBCL cells apoptosis.Re-sults After overexpression of MYD88 L265P,compared with the blank control group(0.670 4±0.017 5)and the negative control group(0.715 3±0.019 6),the MYD88L265P overex-pression group(1.157 2±0.010 2)increased significantly,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(0.69 ±0.04)and the negative control group(0.81±0.07),the MYD88L265P overexpression group(0.48±0.05)was signifi-cantly decreased,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(mRNA:1.0158±0.0115,0.987 3±0.010 2,1.007 6±0.015 3,protein:0.183 4±0.058 9,0.096 8± 0.015 7,0.147 5±0.0418)and negative control group(mR-NA:0.9132±0.0098,1.0032±0.0156,0.9327± 0.011 2,protein:0.187 9±0.042 3,0.088 9±0.0513,0.134 8±0.050 1),the mRNA(3.243 2±0.013 6,2.976 6 ±0.0213,1.585 9±0.019 8)and protein expressions(0.452 7±0.052 4,0.218 9±0.047 5,0.301 4±0.059 8)of IRAK4,NF-κB and anti-apoptosis protein BCL2 in MYD88L265P overexpression group were significantly increased,which was statistically significant(all P＜0.05).Conclusion After overexpression of MYD88 L265P,the apoptosis rate of DLBCL cells decreased and the cell proliferation rate increased.The mechanism may be related to the mutation of MYD88 L265P gene,activation and amplification of NF-κB pathway,and pro-motion of the overexpression of antiapoptotic protein BCL2.

Objective To confirm the willingness of community residents in Xiamen to sign up for hearing care family doctors and its influence factors,and to provide reference for the follow-up promotion of the signing up with family doctors of hearing care.Methods From June to July 2022,a multi-stage stratified sampling method was used to conduct a questionnaire survey among 900 residents from 9 communities in Xiamen.The survey contents in-cluded sociodemographic characteristics,ear and hearing health knowledge and attitude,ear and hearing health be-havior.Chi-square test was used for univariate analysis and multivariate Logistic regression analysis were conducted to analyze the factors affecting the willingness to sign up for family service.Results A total of 900 questionnaires were distributed,and 887 were recared,with an effective return rate of 98.6％.A total of 54.9％(n=487)of the community residents were willing to accept the contracted service of family doctors for hearing health care.Multiva-riate Logistic regression analysis showed that knowledge level of ear and hearing health(OR=0.395,P＜0.05),frequent ear cutting(OR=3.026,P＜0.001)and frequency of otolaryngology(OR=0.399,0.024,P＜0.001)was the influencing factor of community residents'willingness to sign a contract with family doctors for hearing health care.Conclusion Community residents in Xiamen have a high willingness to sign up for family doctor service for hearing health care.Residents with more ear and hearing health knowledge,good ear behavior and medical habits are more willing to sign up for family doctor services for hearing health care.