OBJECTIVE To study the mechanism of action of Rehmannia glutinosa-medicated serum （RG） in regulating lipopolysaccharide （LPS）-induced polarization of BV2 microglia to interfere with neuroinflammation. METHODS An in vitro neuroinflammation model of BV2 cells induced by LPS was established. Cells were divided into control group， LPS group， 10%RG group， 20%RG group， BAY 11-7082 [nuclear factor kappa B （NF-κB） inhibitor] group， and 20%RG+BAY 11-7082 group. Except for the control group， cells in all other groups were pretreated with corresponding drugs for 12 h and then stimulated with LPS for 24 h. The morphology of cells in each group was observed， and the fluorescence intensity of ionized calcium binding adaptor molecule 1 （Iba-1）， inducible nitric oxide synthase （iNOS）， CD206， the mRNA expression of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， IL-1β， iNOS， IL-4， IL-10， transforming growth factor-β （TGF-β）， arginase-1 （Arg-1）， and the Toll-like receptor 4 （TLR4）/NF-κB/NOD-like receptor thermal protein domain associated protein 3 （NLRP3） signaling pathway-related protein expression were detected. RESULTS Compared with the control group， the LPS group showed significant changes in cell morphology， exhibiting a typical M1 activated morphology. The fluorescence intensity of Iba-1 and iNOS， the relative expression levels of TNF-α， IL-6， IL-1β， iNOS mRNA， the relative expression levels of TLR4， NLRP3， caspase-1 p20， IL-1β p17 protein， and the phosphorylation levels of NF-κB p65 and inhibitor of nuclear factor kappa B α were significantly increased （P＜0.05）. The fluorescence intensity of CD206 and the relative expression levels of IL-4， IL-10， TGF-β， and Arg-1 mRNA were significantly reduced （P＜0.05）. Compared with the LPS group， the changes in the above indicators in the cells of the 10%RG group， 20%RG group， BAY 11-7082 group， and 20%RG+BAY 11-7082 group were significantly reversed （P＜0.05）， and there was no statistically significant difference in the comparison of the last three groups （P＞0.05）. CONCLUSIONS RG can effectively inhibit the overall activation of microglia and correct their M1/M2 polarization imbalance， and its mechanism may be closely related to the inhibition of excessive activation of the TLR4/NF-κB/NLRP3 signaling pathway.

BACKGROUND: Currently, lung cancer is one of the malignant tumors with a high morbidity and mortality all over the world. However, the exact mechanisms underlying lung cancer progression remain unclear. The tumor necrosis factor receptor associated factor (TRAF) family members are cytoplasmic adaptor proteins, which function as both adaptor proteins and ubiquitin ligases to regulate diverse receptor signalings, leading to the activation of nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase (MAPK) and interferon regulatory factor (IRF) signaling. The aim of this study was to investigate the expression of TRAFs in different tissues and cancer types, as well as its mRNA expression, protein expression, prognostic significance and functional enrichment analysis in non-small cell lung cancer (NSCLC), in order to provide new strategies for the diagnosis and treatment of NSCLC. METHODS: RNA sequencing data from the The Genotype-Tissue Expression database was used to analyze the expression patterns of TRAF family members in different human tissues. RNA sequencing data from the Cancer Cell Line Encyclopedia database was used to analyze the expression patterns of TRAF family members in different types of cancer cell lines. RNA sequencing data from the The Cancer Genome Atlas (TCGA) database was used to analyze the mRNA levels of TRAF family members across different types of human cancers. Immunohistochemistry (IHC) analyses from HPA database were used to analyze the TRAF protein levels in NSCLC [lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC)]. Overall survival analysis was performed by Log-rank test using original data from Kaplan-Meier Plotter database to evaluate the correlation between TRAF expressions and prognosis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on the TRAF family-related genes using RNA sequencing data from the TCGA database for NSCLC. The correlation between the expression levels of TRAF family members and the tumor immune microenvironment was analyzed using the ESTIMATE algorithm based on RNA sequencing data from the TCGA database. RESULTS: The TRAF family members exhibited significant tissue-specific expression heterogeneity. TRAF2, TRAF3, TRAF6 and TRAF7 were widely expressed in most tissues, while the expressions of TRAF1, TRAF4 and TRAF5 were restricted to specific tissues. The expressions of TRAF family members were highly specific among different types of cancer cell lines. In mRNA database of LUAD and LUSC, the expressions of TRAF2, TRAF4, TRAF5 and TRAF7 were significantly upregulated; while TRAF6 did the opposite; moveover, TRAF1 and TRAF3 only displayed a significant upregulation in LUAD and LUSC, respectively. Except for TRAF3, TRAF4 and TRAF7, other TRAF proteins displayed an obviously deeper IHC staining in LUAD and LUSC tissues compared with normal tissues. Additionally, patients with higher expression levels of TRAF2, TRAF4 and TRAF7 had shorter overall survival; while patients with higher expression levels of TRAF3, TRAF5 and TRAF6 had significantly longer overall survival; however, no significant difference had been observed between TRAF1 expression and the overall survival. TRAF family members differentially regulated multiple pathways, including NF-κB, immune response, cell adhesion and RNA splicing. The expression levels of TRAF family members were closely associated with immune cell infiltration and stromal cell content in the tumor immune microenvironment, with varying positive and negative correlations among different members. CONCLUSIONS TRAF family members exhibit highly specific expression differences across different tissues and cancer types. Most TRAF proteins exhibit upregulation at both mRNA and protein levels in NSCLC, whereas, only upregulated expressions of TRAF2, TRAF4 and TRAF7 predict worse prognosis. The TRAF family members regulate processes such as inflammation, immunity, adhesion and splicing, and influence the tumor immune microenvironment.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/mortality* ; Prognosis ; Gene Expression Regulation, Neoplastic ; Tumor Necrosis Factor Receptor-Associated Peptides and Proteins/metabolism*

AIM:To investigate autophagic status in ischemic stroke with Liver Yang Hyperactivity and the mechanism of Tianma Gouteng Decoction(TMGTD).METHODS:SD rats were divided into sham,model,TMGTD high/medium/low-dose(20.52/10.26/5.13 g·kg-1·d-1),and Nimodipine(30 mg·kg-1·d-1)groups.A Liver Yang Hyperactivity and cerebral ischemia-reperfusion model was es-tablished using Fuzi Decoction(2 g·kg-1·d-1)and thread-occlusion.After 21 days of Fuzi decoction pretreatment,rats received daily drug administra-tion for 12 days.Syndrome indicators(irritability,24-hour water intake,24-hour urine volume,facial temperature)were recorded,plasma NE,E,cAMP,and cGMP were measured by ELISA,neurological function was assessed using Zea Longa and mNSS methods,brain histopathology was evaluated by HE staining,protein expression of soluble/insoluble p62 and LC3B was detected by Western blot,au-tophagy-related genes were analyzed by PCR array,additionally,mRNA and protein levels of CXCR4 and CXCL12 were measured by qRT-PCR and Western blot.RESULTS:Compared to the sham group,the model group showed increased irritability,24-hours water intake,24-hours urine volume,facial temper-ature,and level of NE,E,cGMP(P＜0.01),neurologi-cal scores(P＜0.01),LC3B-Ⅱ,insoluble p62,CXCR4,CXCL12 expression(P＜0.01),but decreased soluble p62(P＜0.01).TMGTD groups exhibited reduced irri-tability,water intake,urine volume,facial tempera-ture,NE,E,cGMP(P＜0.05,P＜0.01),neurological scores(P＜0.05,P＜0.01),p62 expression(P＜0.01),alongside increased LC3B-Ⅱ(P＜0.01)and improved cortical pathology.TMGD also reversed dysregulat-ed autophagy-apoptosis genes(CXCR4,Lamp1,Tgfb1,APP,Rab24)and reduced CXCR4,CXCL12 ex-pression(P＜0.01).CONCLUSION:In the Liver Yang Hyperactivity and cerebral ischemia-reperfusion model,autophagy genes were activated but flux was impaired,and Tianma Gouteng Decoction may protect by restoring autophagic flux and inhibiting the CXCL12/CXCR4 axis.

AIM:To investigate autophagic status in ischemic stroke with Liver Yang Hyperactivity and the mechanism of Tianma Gouteng Decoction(TMGTD).METHODS:SD rats were divided into sham,model,TMGTD high/medium/low-dose(20.52/10.26/5.13 g·kg-1·d-1),and Nimodipine(30 mg·kg-1·d-1)groups.A Liver Yang Hyperactivity and cerebral ischemia-reperfusion model was es-tablished using Fuzi Decoction(2 g·kg-1·d-1)and thread-occlusion.After 21 days of Fuzi decoction pretreatment,rats received daily drug administra-tion for 12 days.Syndrome indicators(irritability,24-hour water intake,24-hour urine volume,facial temperature)were recorded,plasma NE,E,cAMP,and cGMP were measured by ELISA,neurological function was assessed using Zea Longa and mNSS methods,brain histopathology was evaluated by HE staining,protein expression of soluble/insoluble p62 and LC3B was detected by Western blot,au-tophagy-related genes were analyzed by PCR array,additionally,mRNA and protein levels of CXCR4 and CXCL12 were measured by qRT-PCR and Western blot.RESULTS:Compared to the sham group,the model group showed increased irritability,24-hours water intake,24-hours urine volume,facial temper-ature,and level of NE,E,cGMP(P＜0.01),neurologi-cal scores(P＜0.01),LC3B-Ⅱ,insoluble p62,CXCR4,CXCL12 expression(P＜0.01),but decreased soluble p62(P＜0.01).TMGTD groups exhibited reduced irri-tability,water intake,urine volume,facial tempera-ture,NE,E,cGMP(P＜0.05,P＜0.01),neurological scores(P＜0.05,P＜0.01),p62 expression(P＜0.01),alongside increased LC3B-Ⅱ(P＜0.01)and improved cortical pathology.TMGD also reversed dysregulat-ed autophagy-apoptosis genes(CXCR4,Lamp1,Tgfb1,APP,Rab24)and reduced CXCR4,CXCL12 ex-pression(P＜0.01).CONCLUSION:In the Liver Yang Hyperactivity and cerebral ischemia-reperfusion model,autophagy genes were activated but flux was impaired,and Tianma Gouteng Decoction may protect by restoring autophagic flux and inhibiting the CXCL12/CXCR4 axis.

Purpose To investigate the expression and clinical significance of the embryonic lethal abnormal vi-sion-like(ELAVL)family in glioma.Methods Pan-cancer and glioma-specific analyses of mRNA expression profiles of the ELAVL family were analyzed using the TCGA and GTEx databases.The association between ELAVL family ex-pression and survival of glioma patients was evaluated via the gene expression profiling interactive analysis 2(GEPIA2)database.The expression level of ELAVL2 protein in human glioma tissues and non-tumor control brain tissues was ver-ified by immunohistochemistry,and the relationship between its expression and prognosis was analyzed based on the fol-low-up data of patients.Western blot was performed to assess ELAVL2 protein levels in human immortalized astrocytes of UC2 and seven glioma cell lines.Overexpression of ELAVL2 in glioma cells was achieved to evaluate its impact on cell proliferation using in vitro assays.Results Compared to normal tissues,the ELAVL family exhibited distinct ex-pression patterns across various cancers.In glioma,ELAVL1 was significantly upregulated,while ELAVL2,ELAVL3 and ELAVL4 were markedly downregulated.Survival analysis revealed that low ELAVL2 expression was an independent prognostic factor for poor survival in glioma patients(P＜0.05).Immunohistochemistry confirmed that the expression of ELAVL2 decreased with the increase of glioma grade,and its low expression indicated a poor prognosis for patients(P＜0.001).Overexpression of ELAVL2 inhibited glioma cell proliferation in vitro(P＜0.001),suggesting its tumor-suppressive role.Conclusion The ELAVL family members play a critical role in glioma progression.ELAVL2 downregulation serves as a marker for adverse clinical outcomes and represents a potential therapeutic target for glioma therapy.

Purpose To investigate the expression and clinical significance of the embryonic lethal abnormal vi-sion-like(ELAVL)family in glioma.Methods Pan-cancer and glioma-specific analyses of mRNA expression profiles of the ELAVL family were analyzed using the TCGA and GTEx databases.The association between ELAVL family ex-pression and survival of glioma patients was evaluated via the gene expression profiling interactive analysis 2(GEPIA2)database.The expression level of ELAVL2 protein in human glioma tissues and non-tumor control brain tissues was ver-ified by immunohistochemistry,and the relationship between its expression and prognosis was analyzed based on the fol-low-up data of patients.Western blot was performed to assess ELAVL2 protein levels in human immortalized astrocytes of UC2 and seven glioma cell lines.Overexpression of ELAVL2 in glioma cells was achieved to evaluate its impact on cell proliferation using in vitro assays.Results Compared to normal tissues,the ELAVL family exhibited distinct ex-pression patterns across various cancers.In glioma,ELAVL1 was significantly upregulated,while ELAVL2,ELAVL3 and ELAVL4 were markedly downregulated.Survival analysis revealed that low ELAVL2 expression was an independent prognostic factor for poor survival in glioma patients(P＜0.05).Immunohistochemistry confirmed that the expression of ELAVL2 decreased with the increase of glioma grade,and its low expression indicated a poor prognosis for patients(P＜0.001).Overexpression of ELAVL2 inhibited glioma cell proliferation in vitro(P＜0.001),suggesting its tumor-suppressive role.Conclusion The ELAVL family members play a critical role in glioma progression.ELAVL2 downregulation serves as a marker for adverse clinical outcomes and represents a potential therapeutic target for glioma therapy.

In order to investigate how macrophages promote lipid droplet hoarding in hepatocellular carcinoma cells and to delve into the roles of key metabolic enzymes of lipid droplets in the malignant biology of hepatocellular carcinoma cells, the present study was conducted to induce the generation of Hepa1-6 lipid droplets (LDs) using supernatants from tumor-associated macrophages (TAMs), and found that interleukin-10 (IL-10) in TAMs was found to promote the accumulation of lipid droplets. Acetyl-CoA carboxylase alpha (ACC1) is one of the key enzymes for fatty acid synthesis and influences the development of hepatocellular carcinoma. In this study, ACC1 was found to be highly expressed in LDhigh Hepa1-6, and subsequent blockade of ACC1 activity by means of a small molecule inhibitor of ACC1, siRNA interference, and CRISPR-cas9 knockdown was found to reduce the accumulation of Hepa1-6 lipid droplets, as well as to reduce the malignant biological behavior of Hepa1-6 proliferation, and to promote the occurrence of apoptotic events. In summary, IL-10 released by TAMs promoted lipid droplet formation in hepatocellular carcinoma cells, leading to malignant proliferation and apoptosis of hepatocellular carcinoma cells. ACC1 plays a key role in the promotion of lipid droplet accumulation in hepatocellular carcinoma cells by TAMs and may be regulated by IL-10 released by TAMs, and these findings may provide a new target for hepatocellular carcinoma treatment.

Objective　To explore the application of time series models based on meteorological factors in the population density of Aedes albopictus in Hainan Province, and to provide a scientific basis for the prevention and control of dengue fever in Hainan Province. Methods　The density of Aedes albopictus in different habitats in 18 cities and counties of Hainan Province from 2017 to 2022 was monitored monthly using the double-mosquito net trapping index and the Breteau index. Mann-Kendall trend test was used to analyze the temporal trends of the two density indices; Spearman's correlation analysis was employed to assess the correlation strength between each meteorological factor and the two indexes, eliminating unrelated variables, and further selecting the final variables through the full-subset regression method. Three time-series models were constructed for the two density indices, with root mean square error (RMSE), mean absolute error (MAE), and other accuracy metrics used to determine the optimal model; predictions for the density indices for 2023-2024 were made. All statistical analyses were performed in R (4.3.1). Results　The net trapping and Breteau indices showed an overall decrease over the years (Z-values of Mann-Kendall trend test were -6.15 and -4.03, respectively, and P<0.05). The meteorological factor most strongly associated with the trap index was the monthly average minimum temperature; monthly mean minimum temperature and monthly mean relative humidity were strongly correlated with the Breteau index. Based on various evaluation indicators, the multivariate time series model demonstrated the highest accuracy. The study predicts one to two peaks in both the trap index and Breteau Index for the years 2023 and 2024, with peak periods occurring between June to September and May to September, respectively. The predicted value for 2023 aligns with the measured value, demonstrating outstanding predictive accuracy. Conclusions　This study has introduced meteorological factors into the seasonal time series model, allowing for more accurate predictions of the density of Aedes albopictus in Hainan Province from 2023 to 2024, providing a model framework for the prevention and control of dengue fever in Hainan Province.

Objective: To explore the clinical features, the serotype distribution and drug resistance of the isolates in patient with invasive pneumococcal disease (IPD). Methods: By retrieving the laboratory information system in 18 children′s hospitals from 2012 to 2017, the children with IPD were enrolled. Streptococcus pneumoniae (Spn) must be isolated from the sterile sites (blood, cerebrospinal fluid, hydrothorax and joint effusion etc.). The clinical characteristics, serotype, drug resistance, treatment and prognosis were reviewed and analyzed. According to the telephone follow up results, the patients were divided into death group and recovered group. The index as an independent risk factor of mortality was demonstrated by multivariate logistic regression analysis. Results: There were 1 138 children with IPD, including 684 male and 454 female. The proportion of male to female was 1.5∶1. The age ranged from one day to 16 years. The median age was 1 year 3 month. The majority was under 5 years of age (89.3%, n= 1 016), especially under 2 years of age (61.9%, n=704). In all cases, 88.2% (n=1 004) were community acquired infection. The infections included meningitis (n=446, 39.2%), pneumonia with bacteremia (n=339, 29.8%), and bacteremia without focus (n=232, 20.4%). Underlying diseases were found in 242 cases (21.3%). Co-infections were determined in 62 cases (5.4%) with mycoplasma, 27 cases (2.4%) with adenovirus and 34 cases with influenza virus (3.0%). The penicillin insensitivity (PNSP) rates in meningitis and non-meningitis isolates were 69.5% (276/397) and 35.9% (221/615), respectively. There were 81 strains serotyped, in which 93.8% (76/81) were covered by 13-valent protein-polysaccharide conjugate vaccine (PCV13). In the 965 patients who were followed up by phone call, 156 cases (16.2%) were confirmed dead. The independent risk factors for the death were under 2 years of age (OR=2.143, 95%CI 1.284-3.577, P=0.004), meningitis (OR=3.066, 95%CI 1.852-5.074, P<0.01), underlying disease (OR=4.801, 95%CI 2.953-7.804, P<0.01), septic shock(OR=3.542, 95%CI 1.829-6.859, P<0.01), disseminated intravascular coagulation (DIC) (OR=4.150, 95%CI 1.468-11.733, P=0.007), multiple organ failure (OR=12.693, 95%CI 6.623-24.325, P<0.01) and complications of central nervous system (OR=1.975, 95%CI 1.144-3.410, P=0.015). Conclusions Most children with IPD were under 5 years of age, having underlying diseases and acquired the infection in community. The independent risk factors for death were under two years old, meningitis, underlying diseases and multiple organ failure. The problem of drug resistance was severe. The universal immunization of PCV13 would be effective to prevent IPD in Chinese children.

Objective To investigate the effects of thyroid hormone on extremely severe traumatic brain injury (TBI).Methods A retrospective case-control study was conducted to analyze the treatment of 105 patients with extremely severe TBI admitted from July 2010 to April 2014.There were 79 males and 26 females,with an average age of 32.9 years.The patients were divided into conventional treatment group (Group A,35 cases),conventional treatment ± thyroxine treatment group (Group B,35 cases) and thyroxine treatment group after the condition that thyroxine level was low (Group C,35 cases) according to the random number table method.The incidence of low T3 and T4,incidence of hypotension,the dosage of vasoactive drugs,function evaluation of liver and kidney damage,Glasgow outcome scale (GOS),and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) within 20 days after admission,and mortality rate within 30 days after admission were compared and analyzed.Results Within 20 days after admission,the rates of low thyroxine levels and hypotension of the Group B (22.9％,77.1％) were significantly lower than those in the other two groups (Group A:40％,100％;Group C:37％,100％) (all P ＜ 0.05).The doses of dopamine and norepinephrine in Group B was significantly lower than the other two groups and the combination rate of vasopressors in Group B was significantly lower than the other two groups (P ＜ 0.05),while there was no significant difference between Group A and Group C (P ＞ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).The liver and renal dysfunction rates of Group B (29％,31％) were significantly lower than those of the other two groups (Group A:49％,51％;Group C:43％,51％) (all P ＜ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).GOS in Group B [(4.8 ± 1.9) points] was significantly higher than that in Group A [(3.3 ± 0.2) points] (all P ＜ 0.05) within 30 days after admission and significantly higher than that of itself at the beginning [(3.6 ± 1.1) points] (P ＜ 0.05).The APACHE Ⅱ in Group A was significantly higher than those in other two groups as well as that in Group A at admission (P ＜ 0.05).Mortality rates in Group B (31％) and Group C (29％) were significantly lower than that in Group A (69％) within 30 days after admission (P ＜ 0.05).Conclusions Thyroxine can reduce the incidence of hypotension,liver and kidney injury rate in extremely severe TBI.Prevention is better than the supplementary treatment after severe TBI.Thyroxine can also reduce the mortality of extremely severe TBI within 30 days after admission.