Objective:To investigate immunotherapy effects and mechanism of BCG and recombinant BCG(rBCG)with c-di-AMP as adjuvant on melanoma in mice model.Methods:Melanoma mice model was established by B16F10 cell subcutaneous injec-tion in groin,and treated with 1×106 CFU of BCG and rBCG by adjacent injection of subcutaneous tumor for 3 times,respectively.Survival of melanotic mice,tumor growth and metastasis were observed.Tumor tissues of mice were isolated to prepare cell suspen-sion,and proportion of immune cells were detected by flow cytometry.Transcriptional levels of immune-related genes in tumor tissues were detected by qRT-PCR.Results:Both BCG and rBCG immunotherapy could significantly inhibit growth in melanoma mice and prolong survival time of mice.rBCG showed better inhibition on metastasis than BCG.Both strains significantly reduced proportion of M2-type macrophages and myeloid-derived suppressor cell associated with tumor growth and metastasis.Both two strains promoted infiltration of lymphocytes in tumor tissues,and rBCG significantly increased proportion of B cells in tumor.BCG immunotherapy upregulated transcription levels of metastasis-related cytokines,while rBCG therapy had no effects on transcriptions of these genes.Conclusion:Both BCG and rBCG have immunotherapeutic effects on melanotic mice,and rBCG with c-di-AMP as adjuvant shows better inhibition on tumor metastasis than BCG,which mechanism was related to regulation of immune response in tumor tissues.

This study aims to investigate the effect of Lactiplantibacillus plantarum LP12 on obe-sity prevention.In our study,Lactiplantibacillus plantarum LP12 was added to the diet for feed-ing,and the blood biochemistry status of rabbit,as well as the antioxidant effect of serum and liver samples were analyzed by determining the body weight change and feed intake of Japanese White rabbits.The changes in colony structure and abundance were also analyzed by 16S rDNA sequen-cing.The results showed that supplementation of Lactiplantibacillus plantarum LP12 inhibits weight gain,decreases serum glucose and ALT levels,and increases SOD activity in the liver.16S RNA gene sequencing analysis showed that the addition of Lactiplantibacillus plantarum LP12 increases the abundance of Bacteroidetes and Desulfovibrioides at the phylum level,and the supple-mentation of Lactiplantibacillus plantarum LP12 increases the abundance of Muribaculaceae at the genus level.Predictive analysis of microbiota function revealed that the supplementation of Lactiplantibacillus plantarum LP12 positively regulated iron-sulfur clusters and Zn-dependent proteases.In conclusion,the addition of Lactiplantibacillus plantarum effectively inhibits weight gain in Japanese White rabbits,enhances the antioxidative activity of the liver,and induces altera-tions in the gut microbiota composition of these rabbits.These findings lay an experimental foun-dation for further exploring the mechanisms by which Lactobacillus plantarum LP12 exerts its preventive effects against obesity and promotes metabolic health.

Objective To screen metabolic core genes in colon cancer based on machine learning algorithms and analyze their functional mechanisms.Methods Data were obtained from The Cancer Genome Atlas(TCGA)database and the Gene Expression Omnibus(GEO)database.The TCGA co-hort included 375 tumor samples and 32 adjacent normal tissue samples,while the GSE39582 cohort comprised 419 tumor samples.Univariate Cox regression analysis combined with random forest,sup-port vector machine recursive feature elimination(SVM-RFE),and least absolute shrinkage and selec-tion operator(LASSO)regression algorithms were employed to screen for metabolic core genes.Re-ceiver operating characteristic(ROC)curves were plotted,and the area under the curve(AUC)was used to evaluate the predictive efficacy of the core genes.Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)and immunohistochemistry(IHC)methods were adopted to detect the ex-pression of the core genes.The core genes were knocked down to explore their roles in colon cancer.Results Three core genes,namely CPT2,SCP2 and NR3C2,were screened based on machine learning algorithms.According to the comparison results of the AUCs of the ROC curves,NR3C2 exhibited the best predictive efficacy.qRT-PCR detection results showed that NR3C2 mRNA was lowly ex-pressed in colon cancer cell lines;IHC detection results revealed that NR3C2 was lowly expressed in colon cancer tissues.Knocking down NR3C2 significantly promoted the proliferation and migration of colon cancer cells.Conclusion NR3C2 is identified as a core metabolic inhibitory gene in colon cancer by cross-applying three machine learning algorithms,which may provide a new strategy for metabolic targeted therapy.

Adenosine,as a purine nucleoside widely present in living organisms,plays an impor-tant role in such physiological and pathological processes as vasodilation,inflammation regulation,fibrosis,and viral infection.Research has found that in pulmonary hypertension,adenosine acts as a vasodilator through multiple targets.In diseases such as mycoplasma pneumonia,asthma,chronic obstructive pulmonary disease,and idiopathic pulmonary fibrosis,adenosine primarily mediates the progression of inflammation through interactions with adenosine A2A and A2B receptors,thus delivering a dual regulatory effect.This suggests that modulating adenosine receptors is a potential key target for the treatment of pulmonary diseases.Adenosine derivatives-such as acyclovir monophosphate,which has antiviral effects,cyclic adenosine monophosphate,which can alleviate airway narrowing,and N-6-methyladenosine,which is involved in regulating cilia homeostasis and anti-pulmonary fibrosis-are closely related to the occurrence and development of pulmonary diseases.Targeting the adenosine A2A receptor or antagonizing the adenosine A2B receptor can precisely intervene in specific pulmo-nary diseases.Combination medications,regulation of adenosine metabolic pathways,and gene editing technologies promise new treatments.This article reviews the role of adenosine,its receptors,and derivatives in the development of pulmonary diseases in the hopes of providing evidence for related treatment.

Objective:To explore the effects of adenosine on learning and memory in rats thromboembolic stroke(TS)and its mechanism based on PI3K/Akt/CREB signaling pathway.Methods:36 male SD rats were randomly di-vided into sham group(Sham),TS group,TS+adenosine(TS+Ade)group and TS+adenosine+Akt inhibitor(TS+Ade+MK-2206)group.Preparation of rat TS model used autologous thrombus intravascular formation method.Mor-ris water maze(MWM)test was used to observe the learning and memory ability of rats.The morphological changes of hippocampal neurons were observed by Nissl staining.The expression of glutamate transporter 1(GLT-1)and glial fi-brillary acidic protein(GFAP)in CA1 region was determined by immunofluorescence.The protein expressions of PI3K,p-PI3K,Akt,p-Akt,CREB and p-CREB were determined by Western blot.Results:Through the rat TS mod-el,adenosine was able to improve the learning and memory ability,improve the hippocampal neuron cell morphological abpormality and vacuolation,nucleolar condensation and other phenomena in rats.Immunofluorescence results showed that GLT-1 and GFAP were co-localized,and adenosine could significantly enhance the fluorescence intensity of GFAP and GLT-1.Western blot results showed that adenosine can enhance the expression of p-PI3K,p-Akt,and p-CREB proteins,but the use of Akt inhibitors can to some extent inhibit the phosphorylation of PI3K,Akt,and CREB.Conclusion:Adenosine may enhance the expression of GLT-1 in astrocytes and increase the clearance of glutamate through activation of PI3K/Akt/CREB signaling pathway,thereby reducing the excitotoxicity and improving recognition and memory function.However,this effect is partially blocked by Akt inhibitors.

Adenosine,as a purine nucleoside widely present in living organisms,plays an impor-tant role in such physiological and pathological processes as vasodilation,inflammation regulation,fibrosis,and viral infection.Research has found that in pulmonary hypertension,adenosine acts as a vasodilator through multiple targets.In diseases such as mycoplasma pneumonia,asthma,chronic obstructive pulmonary disease,and idiopathic pulmonary fibrosis,adenosine primarily mediates the progression of inflammation through interactions with adenosine A2A and A2B receptors,thus delivering a dual regulatory effect.This suggests that modulating adenosine receptors is a potential key target for the treatment of pulmonary diseases.Adenosine derivatives-such as acyclovir monophosphate,which has antiviral effects,cyclic adenosine monophosphate,which can alleviate airway narrowing,and N-6-methyladenosine,which is involved in regulating cilia homeostasis and anti-pulmonary fibrosis-are closely related to the occurrence and development of pulmonary diseases.Targeting the adenosine A2A receptor or antagonizing the adenosine A2B receptor can precisely intervene in specific pulmo-nary diseases.Combination medications,regulation of adenosine metabolic pathways,and gene editing technologies promise new treatments.This article reviews the role of adenosine,its receptors,and derivatives in the development of pulmonary diseases in the hopes of providing evidence for related treatment.

This study aims to investigate the effect of Lactiplantibacillus plantarum LP12 on obe-sity prevention.In our study,Lactiplantibacillus plantarum LP12 was added to the diet for feed-ing,and the blood biochemistry status of rabbit,as well as the antioxidant effect of serum and liver samples were analyzed by determining the body weight change and feed intake of Japanese White rabbits.The changes in colony structure and abundance were also analyzed by 16S rDNA sequen-cing.The results showed that supplementation of Lactiplantibacillus plantarum LP12 inhibits weight gain,decreases serum glucose and ALT levels,and increases SOD activity in the liver.16S RNA gene sequencing analysis showed that the addition of Lactiplantibacillus plantarum LP12 increases the abundance of Bacteroidetes and Desulfovibrioides at the phylum level,and the supple-mentation of Lactiplantibacillus plantarum LP12 increases the abundance of Muribaculaceae at the genus level.Predictive analysis of microbiota function revealed that the supplementation of Lactiplantibacillus plantarum LP12 positively regulated iron-sulfur clusters and Zn-dependent proteases.In conclusion,the addition of Lactiplantibacillus plantarum effectively inhibits weight gain in Japanese White rabbits,enhances the antioxidative activity of the liver,and induces altera-tions in the gut microbiota composition of these rabbits.These findings lay an experimental foun-dation for further exploring the mechanisms by which Lactobacillus plantarum LP12 exerts its preventive effects against obesity and promotes metabolic health.

Objective:To explore the effects of adenosine on learning and memory in rats thromboembolic stroke(TS)and its mechanism based on PI3K/Akt/CREB signaling pathway.Methods:36 male SD rats were randomly di-vided into sham group(Sham),TS group,TS+adenosine(TS+Ade)group and TS+adenosine+Akt inhibitor(TS+Ade+MK-2206)group.Preparation of rat TS model used autologous thrombus intravascular formation method.Mor-ris water maze(MWM)test was used to observe the learning and memory ability of rats.The morphological changes of hippocampal neurons were observed by Nissl staining.The expression of glutamate transporter 1(GLT-1)and glial fi-brillary acidic protein(GFAP)in CA1 region was determined by immunofluorescence.The protein expressions of PI3K,p-PI3K,Akt,p-Akt,CREB and p-CREB were determined by Western blot.Results:Through the rat TS mod-el,adenosine was able to improve the learning and memory ability,improve the hippocampal neuron cell morphological abpormality and vacuolation,nucleolar condensation and other phenomena in rats.Immunofluorescence results showed that GLT-1 and GFAP were co-localized,and adenosine could significantly enhance the fluorescence intensity of GFAP and GLT-1.Western blot results showed that adenosine can enhance the expression of p-PI3K,p-Akt,and p-CREB proteins,but the use of Akt inhibitors can to some extent inhibit the phosphorylation of PI3K,Akt,and CREB.Conclusion:Adenosine may enhance the expression of GLT-1 in astrocytes and increase the clearance of glutamate through activation of PI3K/Akt/CREB signaling pathway,thereby reducing the excitotoxicity and improving recognition and memory function.However,this effect is partially blocked by Akt inhibitors.

Objective:To investigate immunotherapy effects and mechanism of BCG and recombinant BCG(rBCG)with c-di-AMP as adjuvant on melanoma in mice model.Methods:Melanoma mice model was established by B16F10 cell subcutaneous injec-tion in groin,and treated with 1×106 CFU of BCG and rBCG by adjacent injection of subcutaneous tumor for 3 times,respectively.Survival of melanotic mice,tumor growth and metastasis were observed.Tumor tissues of mice were isolated to prepare cell suspen-sion,and proportion of immune cells were detected by flow cytometry.Transcriptional levels of immune-related genes in tumor tissues were detected by qRT-PCR.Results:Both BCG and rBCG immunotherapy could significantly inhibit growth in melanoma mice and prolong survival time of mice.rBCG showed better inhibition on metastasis than BCG.Both strains significantly reduced proportion of M2-type macrophages and myeloid-derived suppressor cell associated with tumor growth and metastasis.Both two strains promoted infiltration of lymphocytes in tumor tissues,and rBCG significantly increased proportion of B cells in tumor.BCG immunotherapy upregulated transcription levels of metastasis-related cytokines,while rBCG therapy had no effects on transcriptions of these genes.Conclusion:Both BCG and rBCG have immunotherapeutic effects on melanotic mice,and rBCG with c-di-AMP as adjuvant shows better inhibition on tumor metastasis than BCG,which mechanism was related to regulation of immune response in tumor tissues.

Lung-brain interaction refers to the complex physiological and pathological processes in which the lungs and brain influence and regulate each other through various mechanisms. This process works on the complex network between the lung and brain through multiple levels such as nerve, immune, microbial, metabolic, and gas pathways, and affects development of the diseases. This article aims to provide new ideas and methods for prevention and treatment of related lung and brain diseases, as well as references for subsequent research and clinical practice, by reviewing the physiological and pathological mechanisms of neural pathway, immune pathway, and microbial, metabolic and gas pathways in lung brain interaction.