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Objective To construct a human keratinocyte-forming cell line(HaCaT) with stable knockout of the G protein-coupled receptor 107(GPR107) gene, and to preliminarily investigate the effect of GPR107 deletion on the biological behaviour of HaCaT cells. Methods Using CRISPR/Cas9 gene editing technology, HaCaT cells with knockout ofGPR107gene were constructed and monoclonal cells with GPR107 deletion were obtained by limited dilution method. Genomic DNA was amplified using Western blot and PCR and sequenced to validate the single-cell clones with knockdown of GPR107. The cell cycle changes were detected by flow cytometry; cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; changes in cell differentiation markers were detected by Western blot; cell migration ability was analyzed by cell scratch assay and other methods. Results LentiCas9-Blast and plenti-guide-RNA-GPR107 plasmids were successfully transfected into HaCaT cells, 21 monoclonal cell lines were obtained by limited dilution, and Western blot showed that the GPR107 expression was significantly reduced in 8 of them; PCR sequencing of the cellular genome was used, which resulted in the obtainment of C4 and 2D8GPR107-/-HaCaT monoclonal cell lines. CCK-8 assay and flow cytometry assay showed thatGPR107gene deletion resulted in G0G1phase block, significantly weakened proliferation ability and increased apoptosis level of HaCaT cells. Western blot found that the differentiation of HaCaT cells accelerated after knockdown ofGPR107. Additionally the results of the cell scratch assay indicated that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. The results showed that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. Conclusion HaCaT cell line withGPR107gene deletion is successfully constructed, GPR107 deletion blocks the G0G1phase of HaCaT cells, which inhibiting the proliferation of HaCaT cells and promoted apoptosis, and it was found that the differentiation and migration of HaCaT cells were enhanced after knocking downGPR107.

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Objective To investigate the effect of laminarin(LAM) on nonproliferative diabetes retinopathy by high throughput sequencing(RNA-seq). Methods The diabetes model was established by intraperitoneal injection of streptozotocin(STZ), and the effect of LAM on diabetic mice was observed.C57BL/6 mice were randomly divided into three groups: Control group, Model group, and LAM group, with 8 mice in each group. After 8 weeks of modeling, the LAM group received a 4-week intraperitoneal injection of LAM treatment. Changes in blood glucose and body weight of the three groups of mice were recorded, HE staining was performed to examine retinal lesions, and RNA-seq was used to identify differentially expressed genes(DEGs) in diabetic retinopathy(DR) under the action of STZ and LAM. Results STZ successfully established the model of DR, and LAM reduced the blood sugar in diabetic mice to a certain extent and improved the pathological morphology of retinal structural looseness in diabetic mice. After RNA-seq analysis of DEGs, it was found that there were a total of 214 DEGs in the retina of the Model group mice compared to the Control group. Enrichment analysis revealed that DR could exacerbate the lesions through the PI3K Akt signaling pathway. There were a total of 42 DEGs in the retina of the Model group and LAM group mice, and enrichment showed that LAM improved the lesions through the neutrophil extracellular trap pathway. Early growth response factor 1(Egr1), FBJ osteosarcoma oncogene(Fos), nuclear receptor subfamily 4A member 1(Nr4a1), and salt-induced kinase 1(Sik1) were regulated by STZ, and LAM significantly regulated their expression, which might be closely related to LAM′s treatment of diabetic retinopathy. Conclusion DEGs can exacerbate the severity of diabetic retinopathyviathe PI3K-Akt signaling pathway. LAM can mitigate diabetic retinopathyviathe neutrophil extracellular trap pathway. Egr1, Fos, Nr4a1, and Sik1 are key genes involved in LAM treatment of STZ-induced DR.

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Objective To investigate the role of single nucleotide polymorphisms(SNP) of glutathione peroxidase 4(GPX4) gene in the risk of non-cardia gastric cancer. Methods A total of 1 031 samples were selected, including 506 normal examiners and 525 patients with non-cardia gastric cancer.GPX4rs4807542, rs713041, rs2074451 and rs3746162 were genotyped by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Unconditional Logistic regression was used to analyze the relationship between each SNP with the risk of non-cardia gastric cancer under the four genetic models. Results The CT+TT genotype of rs713041 reduced the risk of non-cardia gastric cancer compared with the CC genotype(OR=0.699, 95%CI: 0.537-0.910). The GT+TT genotype of rs2074451 reduced the risk of non-cardia gastric cancer compared with the GG genotype(OR=0.681,95%CI: 0.520-0.893). The G-C-G-C haplotype, constructed by the four SNPs ofGPX4, was related to an increased risk of non-cardia gastric cancer(OR=1.262, 95%CI: 1.035-1.539), and G-T-T-C haplotype was related to a decreased risk of non-cardia gastric cancer(OR=0.784, 95%CI: 0.656-0.937). The fourth-order interaction ofGPX4rs4807542, rs713041, rs2074451 and rs3746162 played a synergistic effect in the risk of non-cardia gastric cancer(P<0.05). Conclusion GPX4rs713041 and rs2074451 are related to non-cardia gastric cancer susceptibility. The G-C-G-C haplotype composed ofGPX4rs4807542, rs713041, rs2074451 and rs3746162 is a risk factor for non-cardia gastric cancer, while the G-T-T-C haplotype is a protective factor. The interaction betweenGPX4rs4807542, rs713041, rs2074451 and rs3746162 is closely connected with the occurrence of non-cardia gastric cancer.

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Objective To investigate the effects of cinnamaldehyde(CA) on the growth, metastasis and apoptosis of human endometriosis(EMs) cells and to explore whether the mechanism is related to ribosomal protein S7(RPS7) expression. Methods Endometriosis cells were divided into control group, CA group, sh-NC group, CA+sh-RPS7 group. Effects of CA on cell growth in human endometriotic cells were determined using Cell Counting Kit-8(CCK-8) and colony formation assay. Effects of CA on cell metastasis were performed by motility assay and Transwell assay. Effects of CA on cell apoptosis were evaluated by Hoechst 33258 staining and flow cytometry. Meanwhile, the levels of PCNA, E-cadherin, Vimentin, Bax and Bcl-2 were evaluated using Western blot in human endometriotic cells with treatment CA. The expression of RPS7 was detected by qRT-PCR and Western blot assay. The RPS7 overexpression of human endometriotic cells was established by cell transfection. CA-mediated effects on cell proliferation and apoptosis were determined by CCK-8 assay and flow cytometry in human endometriotic cells with RPS7 overexpression. Results CA repressed cell growth as well as down-regulated PCNA. The half inhibitory concentration(IC50) value was 53.60 μmol/L after 24 h treatment, and colony formation rate was 25.32%. Additionally, CA inhibited metastasis which was associated with downregulated Vimentin and upregulated E-cadherin. The relative migration rates were 35% and 29% as well as invasion rate was 40%. Further, CA induced apoptosis by cell cycle G2/M phase arrest and cell apoptosis rate was 25.1%, which related to the up-regulation of of Bax and the down-regulation of Bcl-2. CA inhibited the expression of RPS7 and overexpression of RPS7 promoted cell proliferation and suppressed apoptosis in CA-mediated cells. Conclusion CA inhibits cell growth, metastasis, and induces cell apoptosis by downregulating the expression of RPS7.

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Objective To explore the expression level of PPFIA4 in hepatocellular carcinoma tissues and HCCLM3 cells and its regulation of the biological behavior of hepatocellular carcinoma. Methods Bioinformatics analysis, Western blot, and immunohistochemistry were employed to detect the expression of PPFIA4 in tumor tissues of patients with hepatocellular carcinoma and analyze the prognosis of these patients. An siRNA plasmid was designed to knock down the expression of PPFIA4 in HCCLM3 cells. The effects of PPFIA4 knockdown on the migration and invasion abilities of HCCLM3 cells were then evaluated using scratch healing and Transwell assays. Furthermore, Western blot was utilized to detect the expression levels of epithelial-mesenchymal transition(EMT)-related protein markers in the HCCLM3 cell line after transfection with the siRNA plasmid. Results PPFIA4 was highly expressed in hepatocellular carcinoma tissues and hepatocellular carcinoma cells( HCCLM3, Li-7, MHCC97H); the high expression of PPFIA4 indicated that the clinical stage of patients was late and the overall survival(OS) was short. After knocking down the expression of PPFIA4 in HCCLM3 cell line, the migration and invasion ability of HCCLM3 cells decreased(P<0.001) and the expression of EMT markers changed. The expression of epithelial cell marker E-cadherin increased(P<0.01), while the expression of mesenchymal markers Vimentin and N-cadherin decreased(P<0.05,P<0.01). Conclusion PPFIA4 is highly expressed in hepatocellular carcinoma tissues and hepatocellular carcinoma cell lines and is associated with poor prognosis of patients. Silencing PPFIA4 can regulate the biological behavior of hepatocellular carcinoma cells and inhibit the migration and invasion of HCCLM3 cells. The specific mechanism may be related to EMT.

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Objective To clarify the intrinsic link between a high-fat diet(HFD) and the pathological progression and prognosis of spinal cord injury(SCI) while preliminarily exploring the potential underlying mechanisms. Methods SCI models were established in mice that were fed either a regular diet(RD) or HFD, with injury inflicted specifically on the T9-T12segments. Hematoxylin-Eosin(HE) staining, Masson staining, and Nissl staining were used to observe the local histological changes in SCI tissues. The basso, beattie, and bresnahan(BBB) score and footprint analysis were used to evaluate and compare hindlimb functional recovery after SCI in both RD and HFD mice.In vitroexperiments were conducted to identify key fatty acids in the HFD that exacerbate neuronal damage, whilein vivoexperiments assessed the effects of 2-bromopalmitate(2-BP), a palmitic acid inhibitor, on HFD-fed mice with SCI. Results Compared to RD-fed mice, HFD-fed mice exhibited significantly larger lesion areas, more severe neuronal damage, and poorer hindlimb functional recovery after SCI. Palmitic acid was identified as the key fatty acid aggravating neuronal damage. Further more, inhibition of palmitoylation, mediated by palmitic acid, enhanced neuronal survival, promoted tissue repair, and improved hindlimb functional recovery in HFD-fed mice post-SCI. Conclusion HFD exacerbates pathological damage following SCI in mice through palmitic acid, impairing recovery. Palmitic acid-mediated palmitoylation is likely the main mechanism underlying this effect.

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Objective To investigate the expression of LncRNA PXN-AS1(abbreviated as PXN-AS1) in gastric cancer tissues and its impact on the proliferation, apoptosis, and invasion of gastric cancer cells, and to explore its potential regulatory mechanisms. Methods Gastric cancer tissues and adjacent tissues from 43 patients with gastric cancer were collected. The expression levels of PXN-AS1 and miR-125a-5p in gastric cancer tissues were detected using qRT-PCR, and the relationship between the expression level of PXN-AS1 and the clinical pathological parameters of patients with gastric cancer was analyzed. Pearson correlation analysis was conducted to evaluate the correlation between PXN-AS1 and miR-125a-5p in gastric cancer tissues. PXN-AS1 siRNA plasmid(si-PXN-AS1) and negative control plasmid(si-NC), as well as miR-125a-5p inhibitor(inhibitor) and negative control miRNA inhibitor(inhibitor NC), were transfected alone or in combination into HGC-27 cells. The cells were divided into si-NC group, si-PXN-AS1 group, si-PXN-AS1+inhibitor NC group, si-PXN-AS1+inhibitor group, and a blank control group was also established. qRT-PCR was utilized to assess the expression levels of PXN-AS1 and miR-125a-5p in each cell group. CCK-8 assay was employed to evaluate the proliferation levels of cells in each group. EdU experiment was conducted to assess the proliferation status of cells in each group. Transwell assay was performed to examine the migration and invasion capabilities of cells in each group; flow cytometry was employed to measure the apoptosis levels of cells in each group; the dual-luciferase reporter gene system was used to validate the sponge absorption of PXN-AS1 on miR-125a-5p and regulate its expression levels. Bioinformatics analysis was employed to screen downstream target genes of miR-125a-5p and perform GO and KEGG enrichment analysis. Results Compared with adjacent cancerous tissues, the expression level of the PXN-AS1 in gastric cancer tissues significantly increased, while the expression level of miR-125a-5p markedly decreased(bothP<0.05). PXN-AS1 and miR-125a-5p exhibited a negative correlation(r=-0.452,P=0.002). Furthermore, gastric cancer patients with high expression of PXN-AS1 exhibited the proportions of lymph node metastasis positivity and low differentiation were significantly higher(allP<0.05). Compared to the blank group and si-NC group, the si-PXN-AS1 group of HGC-27 cells showed significantly reduced levels of PXN-AS1 expression, cell proliferation capacity, and the number of cell migration and invasion(allP<0.05), while the level of cell apoptosis significantly increased(P<0.05). The dual-luciferase reporter system results demonstrated specific adsorption of miR-125a-5p by PXN-AS1 in HGC-27 cells. Knockdown of miR-125a-5p could reverse the effect of PXN-AS1 silencing on the inhibition of proliferation, migration and invasion of gastric cancer cells, and promote apoptosis. The bioinformatics analysis results indicated that the downstream target genes of miR-125a-5p were involved in various biological processes, including DNA damage, T cell apoptosis and differentiation, RNA metabolism synthesis processes, and signal transduction. Additionally, miR-125a-5p was implicated in signaling pathways such as the HIF-1 pathway, JAK-STAT pathway, and cell apoptosis signaling pathway. Conclusion PXN-AS1 is upregulated in gastric cancer. Moreover, acting as a competitive sponge for miR-125a-5p, PXN-AS1 promotes the proliferation, migration, and invasion abilities of gastric cancer cells HGC-27 while inhibiting apoptosis.

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Objective To investigate the therapeutic effects and underlying mechanisms of cannabidiol(CBD) and its nano-formulations on depression-like behaviors in mice. Methods A murine model of acute anxiety and depression was established by intraperitoneal administration of lipopolysaccharide(LPS). A total of 55 mice were randomly assigned into several groups: for the long-term study, a control group(Con), a model group(LPS), a cannabidiol group(CBD), a nano-cannabidiol group(NCBD), and a sertraline(SER) group, each consisting of 7 mice. In the short-term study, mice were divided into four groups: the Con group, LPS group, CBD group, and NCBD group, with 5 mice in each group. Except for the Con group and LPS group, which were given distilled water, the remaining groups were administered 25 and 50 mg/kg of cannabidiol and its nano-formulationviaoral gavage. The open field and forced swimming tests were employed to assess anxiety-and depression-like behaviors inmice. Luxol Fast Blue myelin staining was employed to evaluate myelin sheath morphology in the prefrontal cortex, and immunofluorescence staining was utilized to quantify the protein expression levels of silencing information regulator(SIRT2), ionized calcium binding adaptor molecule-1(Iba-1), and interleukin-1β(IL-1β) in the prefrontal cortex. Results In the long-term experiment, the LPS group exhibited a significant reduction in shuttle times(P<0.05), an increase in immobility time(P<0.01), and a decrease in the number and length of myelin sheaths(P<0.05) compared to the Con group. Compared to the LPS group, the depressive behaviors in the CBD, NCBD, and SER groups were significantly alleviated(P<0.01), and the number and length of myelin sheaths increased(P<0.05). In the short-term experiment, compared to the Con group, the LPS group exhibited significantly increased anxiety-and depression-like behaviors(P<0.05), downregulated SIRT2 expression(P<0.01), and upregulated Iba-1 and IL-1β expression(P<0.01). The CBD and NCBD groups demonstrated a reduction in anxiety and depression-like behaviors(P<0.05), an increase in SIRT2 expression(P<0.01), and a decrease in Iba-1 and IL-1β expressions(P<0.05) compared to the LPS group. Conclusion CBD and its nano-formulations effectively mitigate anxiety and depression-like behaviors in mice. The underlying mechanisms may be associated with the reversal of SIRT2 protein expression, demyelination changes, microglial activation, and the levels of inflammatory factors in the prefrontal cortex.

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Objective To investigate the role of caspase recruitment domain-containing protein 9(Card9) inAspergillus fumigatus(A.fumigatus) keratitis and the effect of its deficiency on macrophage resistance to fungal infection. Methods (1) C57BL/7 mice aged 6-8 weeks were selected and the mice pretreated Card9 siRNA and Blank siRNA, respectively, and the expression of Card9 in each group was detected by Western blot and RT-PCR. The corneal epithelium of the mice was scraped away 72 hours later, andA.fumigatusspore suspension was injected into the corneal stroma. The corneal scores were recorded at 1 d, 3 d, 5 d and 7 d after infection. The expression of Card9, nuclear factor κB(NF-κB), interleukin 1β(IL-1β), interleukin 6(IL-6) and tumor necrosis factor α(TNF-α) in each group was detected by RT-PCR and immunohistochemical(IHC).(2) Human corneal epithelial cells(HCECs) and human monocytic-leukemia cells(THP-1)in vitro, RT-PCR was used to examine the expression of Card9 gene in the two cells, and a stable cell line of THP-1 cells was constructed using shRNA vectors. The expression of Card9 in the cell line was detected by Western Blot and RT-PCR. The cells were induced into macrophages and stimulation byA.fumigatus, and the expression of Card9, NF-κB, IL-1β, IL-6 and TNF-α was detected by RT-PCR. Results Card9 expression increased inA.fumigatuskeratitis, mainly distributed in cytoplasm of immune cells. The expression of Card9 in the cornea of mice treated with Card9 siRNA was significantly reduced. After inhibiting the expression of Card9 gene, the expressions of Card9, NF-κB, IL-1β, IL-6 and TNF-α significantly decreased and the changes of IL-1β were most significant. Inin vitrostudies, Card9 exhibited negligible expression in human corneal epithelial cells, contrasting with its pronounced expression in THP-1 cells. After the induction of macrophages, Card9, NF-κB, IL-1β, IL-6, and TNF-α were significantly upregulated under the stimulation ofA.fumigatus. After inhibiting the expression of Card9, the stimulated expression of these factors was significantly reduced, with the most notable change observed in IL-1β. Conclusion Card9 is involved in the inflammatory development and healing process ofA.fumigatuskeratitis. Card9 deficiency can cause functional impairment of macrophages and inhibit the expression of inflammatory factors to a certain extent, in which IL-1β has the greatest effect.

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Objective To investigate the effects of usnic acid(UA) on proliferation, cell cycle, apoptosis and autophagy of lung cancer cells A549 and NCI-H358. Methods The CCK-8 method was used to detect the inhibitory effect of UA on two kinds of lung cancer cells proliferation. Flow cytometry was used to detect the cell cycle arrest effect of UA on two types of lung cancer cells. The fluorescence amount of UA-induced reactive oxygen species(ROS) in two kinds of lung cancer cells were detected by DCFH-DA probe assay and flow cytometry. Western blot was used to detect the expression of apoptosis related proteins Bax, Caspase-3, Cleaved-Caspase-3, and autophagy related proteins LC3-Ⅰ and LC3-Ⅱ in two types of lung cancer cells after treatment with UA and UA+ROS inhibition. Results(1) Usnic acid reduced the survival rates of two types of cells and had a stronger ability to inhibit the proliferation of A549 cells than NCI-H358 cells.(2) Usnic acid blocked A549 cells in the G0/G1phase, while NCI-H358 cells in the G2/M and S phases.(3) Usnic acid induced an increase in ROS content in two types of cells. Compared to A549, NCI-H358 cells showed a greater increase in ROS, and the ROS inhibitor reduced the intracellular ROS increase induced by UA.(4) Usnic acid induced high expression of apoptosis-related proteins Bax and Cleaved Caspase-3 and increased the ratio of autophagy-related proteins LC3-Ⅱ/LC3-Ⅰ in both lung adenocarcinoma cells, and its pro-apoptotic and autophagic effects were stronger in A549 than in NCI-H358. After ROS inhibition, the expression levels of Bax and Cleaved Caspase-3 and the LC3-Ⅱ/LC3-Ⅰ ratio of both lung adenocarcinoma cells decreased, and the decrease was greater in NCI-H358 cells. Conclusion Usnic acid inhibits the proliferation of lung cancer A549 and NCI-H358 cells, induces cell cycle arrest, and induces apoptosis and autophagy in cancer cells. The inhibitory and killing effect of UA on A549 cells is stronger than that on NCI-H358 cells. In this case, the induced cell ROS are involved in the action of UA in two types of lung cancer cells. In contrast to A549 cells,ROS might play a more significant role in mediating the apoptosis and autophagy induced by usnic acid in NCI-H358 cells.

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Objective To establish a PTEN gene-downregulated ASD juvenile rat model (VPA-ADV) through combined application of valproic acid (VPA) and PTEN adenovirus (ADV),then to compare the newly constructed animal model with two traditional autistic animal models of VPA and AD,and ultimately to prove the advantages of this model in animal model establishment of acupuncture treatment for ASD.Methods Wista rats at 12.5 days of gestation were randomly divided into 2 groups.VPA rats were given 600 mg/kg of normal saline (NS) intraperitoneally.Weight,eye opening time and tail deformity were recorded.The newborn mice in NS group were randomly divided into three groups (10 rats in each group):normal (NS) group,virus (ADV) group and virus-negative control (ADV-NC) group;VPA group (20 young rats) was randomly divided into 2 groups (10 rats in each group):valproic acid (VPA) group and valproic acid-binding virus interference (VPA+ADV) group.The body weight,tail length,curved tail,geotaxis text time and skeletal deformity of 5 groups of young rats after birth,the neurobehavioral behavior of 21-day-old rats,and the myelin structure of brain tissue under electron microscope were observed,the expression levels of PTEN,PI3K,AKT,GSK3β and MBP were detected by immunohistochemistry,Western blot and q-PCR.Results Compared with the NS group,the VPA group had significantly increased malformation rate,slow weight gain,slow tail length growth,and long negative geotaxis reflex time (P<0.05).Compared with the NS group,the weight gain,tail length growth and negative geotaxis reflex time of the three model groups were slower (P<0.05),and the performance of VPA+ADV model was the most significant.There were significant differences in the time of crossing the central grid,the number of crossing the edge grid,and the time of crossing the edge grid in the open field test between the three groups and the NS group (P<0.05).In the selfgrooming experiment,the number of interactions in VPA-ADV model was the least (P<0.05),and the number of digging and self-grooming was the most (P<0.05).In the three-box social experiment,VPA-ADV model had the shortest average number of entries into the social box and the shortest residence time (P<0.05),the time of finding the platform in the water maze experiment was the longest,and the number of times crossing the third quadrant was the least (P<0.05).The structure of the myelin sheath layer in the corpus callosum was observed by transmission electron microscopy.The structure of the NS group was clear and complete.Compared with the NS group,the myelin structure of the ADV-NC group was similar,and the myelin structure of the three model groups was stratified and broken,and there were pathological changes and myelin damage in the ASD.Among them,the myelin sheath of the VPA-ADV model was thickened,stratified,and severe visible disintegration.The results of immunohistochemistry,Western blot and q-PCR showed that the expression of PTEN in VPA+ADV model was down-regulated by about 50%,which was the most obvious.The expression of AKT and MBP increased,and the expression of GSK3β decreased (P<0.05).However,the results of q-PCR showed that the expression of PI3K-mRNA in the three model groups significantly increased (P<0.05),and the change of VPA+ADV model was the most significant.Conclusion The novel PTEN-ASD animal model established by VPA+ADV method is observed to have significant pathological changes that are typical of the manifestation of ASD myelin dysplasia and is determined to have better results than the two traditional autistic animal models.

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Objective To investigate the effects and underlying mechanisms of high mobility group nucleosome-binding domain protein 2(HMGN2) on lung adenocarcinoma cells. Methods This work first analyzed the association between HMGN2 and lung adenocarcinoma tissues using The Cancer Genome Atlas(TCGA) database. Lung adenocarcinoma tissues and adjacent normal tissues were collected to compare the differential expression levels of HMGN2. The expression of HMGN2 mRNA in lung adenocarcinoma cell lines A549 and NC-H1299 were detected by qRT-PCR and Western blot. HMGN2 expression was knocked down using si-RNA technology, with the control group transfected with an equivalent amount of NC-siRNA, and the si-RNA group transfected with si-HMGN2. Stable transfected cell lines were established based on si-RNA knockdown efficiency. The effects of HMGN2 knockdown on the growth, movement, and spread of lung adenocarcinoma cells were assessed using CCK-8, Transwell assays, scratch assays, colony formation assays, and EdU assays. Transcriptome sequencing analysis revealed pathways related to tumorigenesis associated with HMGN2. The relative expression levels of MAPK pathway proteins after HMGN2 knockdown were detected by Western blot. Results HMGN2 mRNA expression was significantly elevated in lung cancer tissues and lung adenocarcinoma cell lines(P<0.05). After HMGN2 knockdown, cell proliferation, migration, and invasion were significantly reduced(P<0.05), and the phosphorylation levels of the MAPK signaling pathway markedly decreased(P<0.05). Conclusion HMGN2 enhances the proliferation, migration, and invasion of lung adenocarcinoma cells, and its mechanism may be closely related to the activation of the MAPK signaling pathwayviaphosphorylation.

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Objective To investigate the expressions of GPR15 and FOXP3 in colorectal carcinoma(CRC) tissues and their clinical prognostic values. Methods A total of 132 patients with CRC underwent radical surgery were collected. The control group selected the normal mucosal tissues more than 5 cm away from the edge of the cancer focus. Immunohistochemistry(Envision two-step method) was used to detect the expression levels of GPR15 and FOXP3 in CRC and adjacent tissues, and analyze their relationships with clinicopathological factors of colorectal cancer. Kaplan-Meier method was used to draw the survival curve to analyze the correlation between the expressions of GPR15 and FOXP3 and the survival prognosis of patients with CRC. The factors influencing prognosis of patients with colorectal cancer were analyzed by Cox regression. Results The immunohistochemistry showed that the expression levels of GPR15 and FOXP3 in CRC were significantly higher than those in normal colorectal mucosal tissues(P<0.05). The expression of GPR15 in CRC tissues was correlated with location, nerve invasion and TNM stage; FOXP3 expression was correlated with sex(P<0.05).Both expressions were not significantly correlated with the clinicopathologic features of age, tumor size, differentiation degree, tissue type, depth of invasion, tumor budding, vascular invasion and lymph node metastasis. Correlation analysis showed that there was no significant correlation between GPR15 and FOXP3 expression(Kappa=-0.019,P>0.05). The survival prognosis of GPR15 positive group was significantly worse than that of negative group(log-rank: χ2=4.3,P=0.039);while the survival prognosis of FOXP3 positive group was significantly better than that of negative group(log-rank: χ2=7.3,P=0.007).Age ≤55 years, positive GPR15 and negative FOXP3 were independent risk factors for poor prognosis in patients with CRC(P<0.05). Conclusion The expression levels of GPR15 and FOXP3 in CRC are significantly higher than those in paracancer tissues, GPR15 and FOXP3 are expected to become new tumor markers for early screening, accurate treatment and prognosis assessment of CRC.

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Objective To explore whether chrysophanol(CHR) affects macrophage polarization by promoting mitochondrial biosynthesis through AMPK/PGC-1α pathway. Methods The molecular docking and binding ability of CHR with AMPK and PGC-1α were predicted by Autodock vina software. Human monocytes(THP-1) were induced to M0 macrophages by phorbol myristate acetate(PMA), and to M1 macrophages by lipopolysaccharide(LPS) combined with interferon-γ(IFN-γ), which were set as Control group. M1 macrophages treated with CHR were set as CHR group. M1 macrophages treated with CHR combined with AMPK inhibitor(Compound C) were set as CHR+Compound C group. The mRNA expression levels of M1 macrophage markers(iNOS, CD86) and mitochondrial biosynthesis related genes(PGC-1α, NFR-1, TFAM) were detected by Quantitative real time polymerase chain reaction(qRT-PCR). The expression level of M1 macrophage marker iNOS was detected by immunofluorescence. The protein expression levels of AMPK, p-AMPK and PGC-1α were detected by Western blot. Results The docking results showed that the binding energies of CHR with AMPK and PGC-1α were-8.4 kcal/mol and-7.4 kcal/mol, respectively. qRT-PCR results showed that the in vitro model of M1 macrophages was successfully established. Compared with the Control group, CHR treatment significantly increased the mRNA expression of mitochondrial biosynthesis-related genes PGC-1α, NFR-1, and TFAM(P<0.001). Compared with CHR treatment group, CHR combined with Compound C treatment significantly decreased the mRNA expression levels of mitochondrial biosynthesis-related genes PGC-1α, NFR-1, and TFAM(P<0.05). Immunofluorescence results showed that CHR treatment inhibited the protein expression of iNOS compared with the Control group(P<0.001). Compared with CHR treatment group,CHR combined with Compound C treatment reversed the inhibitory effect of CHR on i NOS protein expression(P<0.05). Western blot results showed that compared with the Control group,the CHR treatment group had significant increase in the protein expression levels of p-AMPK and PGC-1α(P<0.001).Compared with CHR treatment group,CHR combined with Compound C treatment significantly decreased the protein expression levels of p-AMPK and PGC-1α(P<0.05). Conclusion Chrysophanol may inhibit macrophage polarization to M1 by activating AMPK/PGC-1α signaling pathway to promote mitochondrial biosynthesis.

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Objective To investigate the effects of branched-chain amino acid(BCAA) on the differentiation of 3T3-L1 preadipocytes and its potential mechanism. Methods 3T3-L1 preadipocytes were divided into the Control, differentiation medium(DM), low-concentration BCAA, and high-concentration BCAA groups. A CCK-8 assay was utilized to evaluate pre-adipocyte survival under various BCAA concentrations. Oil-red O staining was used to observe the formation of lipid droplets in adipocytes. Intracellular triglyceride(TG) and total cholesterol(TC) were detected by enzymatic method. RT-qPCR and Western blot were used to detect the mRNA and protein expression of Stat3 and adipocyte differentiation-related genes. Results CCK-8 results showed that the viability of 3T3-L1 cells was not affected when the BCAA concentration was ≤ 10 mmol/L. Compared with the DM group, the low-concentration BCAA groups(0.5 and 1.0 mmol/L) had significantly larger intracellular lipid droplets, increased number of lipid droplets, and elevated levels of the intracellular TC(0.88vs0.68 mmol/g; 0.83vs0.68 mmol/g,P<0.01) and TG(0.77vs0.40 mmol/g; 0.62vs0.40 mmol/g,P<0.01). Nevertheless, the cell differentiation in the high-concentration group(5.0 and 10.0 mmol/L) significantly decreased compared with that in the DM group. Further, levels of PPARγ, C/EBPα, Adiponectin, and FABP4 mRNA and protein expression significantly increased in the low-concentration group, but significantly decreased in the high-concentration group than that in the DM group(P<0.01). In addition, low concentrations of BCAA promoted stat3 phosphorylation, while high concentrations inhibited its phosphorylation(P<0.01). Conclusion BCAA have a dual role in regulating the differentiation of preadipocytes through Stat3, i.e. low concentrations of BCAA induce cell differentiation by promoting Stat3 phosphorylation; whereas high concentrations of BCAA inhibit Stat3 phosphorylation and cell differentiation.

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Objective To monitor changes in blood pressure during rapid eye movement(REM) and non-REM(NREM) sleep in patients with obstructive sleep apnea(OSA), and to analyze the effect of obstructive respiratory events on blood pressure during different sleep stages. Methods A cross-sectional trial was adopted on 1 219 patients diagnosed with OSA. The obstructive respiratory events and blood pressure of patients during REM and NREM sleep were monitored. Additionally, the symptoms of blood pressure fluctuations in OSA patients during different sleep periods and the effects of obstructive respiratory events on blood pressure fluctuations were analyzed. Results SBP and DBP during REM and NREM sleep both tended to rise as OSA worsened. The differences in SBP, DBP, mean apnea time(MAT), mean hypoventilation time(MHT), lowest oxygen saturation(LSpO2), and oxygen desaturation index(ODI) in different sleep periods were statistically significant(P<0.05). Multiple linear regression analysis revealed that SBP during REM sleep was negatively correlated with LSpO2and the apnea-hypopnea index(AHI)(P<0.05), but positively correlated with ODI(P<0.05). DBP during REM sleep was negatively correlated with LSpO2, AHI, and MHT(P<0.05), but positively correlated with ODI(P<0.05); SBP and DBP during NREM sleep were negatively correlated with LSpO2(P<0.05). Conclusion As the severity of OSA increases, the blood pressure values during different sleep periods rise accordingly. The blood pressure levels during REM sleep in OSA patients are higher than those during NREM sleep. LSpO2is an important factor causing blood pressure changes in different sleep periods. AHI, ODI, and MHT are important factors causing blood pressure changes during REM sleep.

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Objective To explore the feasibility of machine learning methods in the discrimination of tuberculosis patients. Methods The data of 15 observation indicators of 860 patients were obtained from a tertiary hospital. Through in-depth mining and analysis of the data, support vector machine, random forest and neural network model methods were used to discriminate the diseases of patients. Results The accuracies of the TB suspected patient discrimination models based on support vector machine, random forest and neural network were 90%, 91% and 88%, respectively. Conclusion All three machine learning methods can be used for discriminative analysis of suspected tuberculosis patients. In comparison, random forest performs better in discriminating patients with tuberculosis from those with pneumonia.

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Objective To construct and validate a clinical diagnostic model to differentiate between pulmonary tuberculosis and non-tuberculous lung diseases. Methods Information was collected from 258 patients with respiratory system diseases, and they were divided into a training set of 152 cases and a test set of 106 cases with a ratio of 6 ∶4 using the random number seed method in R software. The training set was further divided into a tuberculosis group of 95 cases and a non-tuberculosis group of 57 cases, and the test set into a tuberculosis group of 65 cases and a non-tuberculosis group of 41 cases based on the diagnosis of pulmonary tuberculosis. A diagnostic model was constructed using multivariate logistic regression analysis to determine the influencing factors of pulmonary tuberculosis. The diagnostic value and clinical utility of the model were assessed using the receiver operating characteristic(ROC) curve, calibration curve, and decision curve analysis(DCA). Results CD161+%(OR=0.768; 95%CI0.697-0.845;P<0.001), AST(OR=0.961; 95%CI0.930-0.993;P=0.019), and smoking history(OR=3.181; 95%CI1.149-8.804;P=0.026) were identified as independent risk factors for the occurrence of pulmonary tuberculosis. In both the training and test sets, the area under the ROC curve(AUC) reached 0.870(95%CI0.816-0.924) and 0.887(95%CI0.827-0.948), respectively. The Hosmer-Lemeshow goodness-of-fit test showed a good fit(training set χ2=6.213,P=0.623; test set χ2=6.197,P=0.625). DCA indicated that the model had good reference significance for the diagnosis of the probability of pulmonary tuberculosis occurrence. Conclusion The diagnostic model constructed using the percentage of CD161+%, AST levels, and smoking history has certain diagnostic performance, facilitating rapid clinical differentiation between pulmonary tuberculosis and non-tuberculous lung diseases.

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Objective To investigate the prevalence of negative emotions in hospitalized youth patients with type 2 diabetes(T2DM) and its correlation with coping strategies and psychological resilience. Methods 141 youth T2DM patients who met the research standards were selected. Blood glucose related indicators, blood pressure, body mass index(BMI), diabetes chronic complications screening results and other data were collected. The basic information and disease related information questionnaire, self-rating depression scale(SDS), self-rating anxiety scale(SAS), diabetes distress scale(DDS), medical coping modes questionnaire(MCMQ) and Connor-Davidson resilience scale(CD-RISC) were completed. Results Among 141 hospitalized youth T2DM patients, 37.6% were combined with depression, 32.6% were combined with anxiety, and 35.5% were combined with diabetic distress(DD). Univariate analysis showed that systolic blood pressure(P<0.01), educational level, and the form of hospitalization expenses(P<0.05) were significantly correlated with depression. Marital status(P<0.01), family residence, blood glucose monitoring methods, and the last fasting blood glucose(P<0.05) were significantly correlated with anxiety. BMI, whether it was first diagnosed or treated(P<0.01), gender, occupation, disease course, weekly blood glucose monitoring frequency, and the presence of chronic complications(P<0.05) were significantly correlated with DD. In multivariate analysis, systolic blood pressure(P<0.01), educational level, and the form of hospitalization expenses were significantly correlated with depression, marital status(P<0.05) was significantly correlated with anxiety; BMI and weekly blood glucose monitoring frequency(P<0.01) were significantly correlated with DD. SDS, SAS, total scores and dimensions of DDS were negatively correlated with the total score and dimensions of CD-RISC(rs=-0.182--0.467, P<0.05 or 0.01), and positively correlated with the yielding coping strategies(rs=0.177-0.271,P<0.05 or 0.01). SAS,total scores and dimensions of DDS were positively correlated with avoiding coping strategies(rs=0.237-0.419,P<0.05 or 0.01). The total and dimensions of CD-RISC were positively correlated with facing coping strategies(rs=0.215-0.349,P<0.05 or 0.01),and negatively correlated with yielding coping strategies(rs=-0.234--0.325,P<0.01). Conclusion More than 30% of hospitalized youth T2DM may experience negative emotions such as depression,anxiety,and DD. The occurrence of negative emotions in such patients may be related to disease management or socio-economic issues such as systolic blood pressure,educational level,hospitalization expenses,marital status,BMI,and frequency of blood glucose monitoring,as well as decreased psychological resilience and negative coping strategies.

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Objective To investigate the diagnostic value of plasma calprotectin in childern with refractory mycoplasma pneumoniae pneumonia(RMPP). Methods A multicenter, prospective cohort study was conducted, enrolling 228 children with mycoplasma pneumoniae pneumonia(MPP). Among these, 177 cases were diagnosed with general mycoplasma pneumoniae pneumonia(GMPP), while the remaining 51 cases were RMPP. Plasma was collected at the time of admission of the children in both groups, and calprotectin levels were measured. A one-way difference analysis was performed on the blood test indexes of the children in the two groups, and the difference variables withP<0.05 between the two groups were included in a multifactorial logistic regression to analyze the risk factors for the progression of GMPP to RMPP. The differential diagnostic value of plasma calprotectin for GMPP and RMPP was analyzed by the receiver operating characteristic(ROC) curves. Results Univariate analysis showed that plasma calprotectin levels were significantly higher in the RMPP group than those in the GMPP group, and the difference was statistically significant(P<0.05). Logistic regression analysis showed that plasma calprotectin was an independent risk factor for RMPP(OR=1.323,P<0.001), ROC curve analysis showed that plasma calprotectin had a higher diagnostic value for the differential diagnosis of GMPP and RMPP(AUC =0.839), and its combination with C-reactive protein and albumin could significantly improve the diagnostic efficiency. Conclusion Plasma calprotectin has good clinical value for the diagnosis of RMPP.

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Objective To analyze the current status of anti-PD-1 combined with chemotherapy in advanced gastric cancer. Methods The clinical data of 120 patients with advanced gastric cancer treated with chemotherapy or anti-PD-1 plus chemotherapy were retrospectively analyzed. The efficacy [measures include objective response rate(ORR), disease control rate(DCR), and progression-free survival(PFS)]and the occurrence of adverse reactions in patients was monitored. Univariate and multivariate Cox were used to analyze the relationship between clinicopathologic data [including age, gender, body mass index(BMI), differentiation degree, and related hematological indexes], specific treatment conditions and PFS, and to analyze the influence of treatment plan and line number on the curative effect. Results The PFS of chemotherapy plus immunotherapy was significantly better than that of chemotherapy alone(P<0.05), and the median PFS of the two was 7.17 months(95%CI5.85-8.49) and 5.33 months(95%CI3.96-5.70), respectively. In the combination group, the overall ORR was 27.5% and DCR was 94.8%. In the chemotherapy group alone, the overall ORR was 9.7% and DCR was 69.4%.In the chemotherapy plus anti-PD-1 group, a high prognostic nutritional index(PNI) level also provided superior PFS in patients with a 47% reduction in the risk of progression versus a low PNI level [HR0.53(95%CI0.30-0.96);P<0.05]. The median PFS was 7.47 months(95%CI5.83-9.11) with a high PNI level versus 6.00 months(95%CI4.45-7.55) with a low PNI level. The overall incidence of adverse reactions of chemotherapy combined with anti-PD-1 was slightly higher than that of chemotherapy alone, and the incidence of adverse reactions of grade 3-4 was similar, within a controllable range, and could be improved after clinical treatment. Conclusion The efficacy of anti-PD-1 combined with chemotherapy in patients with advanced gastric cancer is better than that of chemotherapy, and the adverse reactions are controllable. In the chemotherapy plus anti-PD-1 group, patients with high PNI levels achieved a higher PFS.

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Objective To explore the differences in oxidative stress levels between patients with and without non-suicidal self-injury(NSSI) behavior in depressive disorders, and their correlation with depressive symptoms, disease duration, and self-injury frequency.Methods A total of 95 subjects were included in the study, including 65 patients with depression(patient group). They were divided into NSSI subgroup(31 cases) and Non-NSSI subgroup(34 cases) based on whether they had NSSI. 30 healthy control group cases(HC). Enzyme linked immunosorbent assay(ELISA) was used to detect the levels of superoxide dismutase(SOD), catalase(CAT), malondialdehyde(MDA), glutathione peroxidase(GPX), and oxidized low-density lipoprotein(OxLDL) in the peripheral blood of the subjects. The Hamilton depression rating scale(HAMD-17) was used to assess the patient′s condition, and the Ottawa self-injury inventory(OSI) was used to quantitatively evaluate the self-injury frequency of NSSI group patients.Results The levels of SOD, CAT,MDA,GPX and OxLDL in the patient group were significantly higher than those in the control group(P<0.05). The levels of SOD, CAT, MDA,GPX and OxLDL in the NSSI subgroup were significantly higher than those in the Non-NSSI subgroup and the control group(P<0.05).The duration of the NSSI subgroup was positively correlated with the levels of SOD,CAT,and MDA(r=0.504,P=0.004; r=0.568,P=0.001; r=0.404,P=0.024),and the HAMD-17 score was positively correlated with the levels of SOD,CAT,and MDA(r=0.573,P=0.001; r=0.577,P=0.001; r=0.477,P=0.007). The frequency of self-injury was positively correlated with the levels of SOD and MDA(r=0.558,P=0.001; r=0.494,P=0.005). Conclusion The OS level of patients with depression is significantly higher than that of the healthy population,and among them,patients with depression accompanied by NSSI have a more significant increase in oxidative stress level. Elevated levels of OS may affect the severity,duration,and frequency of self-injurious behavior in patients with depression accompanied by NSSI.

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Objective To analyze the correlation between different laboratory biomarkers and disease activity in ankylosing spondylitis and to compare their specificity and sensitivity in assessing disease activity. Methods Spearman correlation or Pearson correlation was used to analyze the correlation between disease activity and laboratory biomarkers. Receiver operating characteristic(ROC) was used to compare the sensitivity and specificity of each laboratory biomarker in evaluating disease activity. Results Hypersensitive C-reactive protein, fibrinogen, D-dimer, erythrocyte sediment rate, C-reactive protein, immuno-inflammatory index(platelet count×neutrophil count/lymphocyte count), fibrinogen/albumin ratio, albumin and pro-albumin were correlated with disease activity. The ratio of fibrinogen to albumin, fibrinogen, erythrocyte sedimentation rate, immuno-inflammatory index, C-reactive protein and hypersensitive C-reactive protein had good values in determining the disease activity. Conclusion Different laboratory biomarkers are correlated with the disease activity of ankylosing spondylitis, and some of them have better discriminating values for the disease activity.

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Objective To explore the risk factors of prognosis in patients with severe community-acquired pneumonia(SCAP) in different age groups. Methods A multi-center and prospective study was conducted at 11 teaching hospitals in China from December 2017 to October 2021. Patients who met the criteria were assigned to the elderly group(≥65 years) and the non-elderly group(18-64 years) to demonstrate the clinical characteristics of SCAP. Patients were divided into survival group and death group according to whether they died in hospital, to determine the risk factors associated with mortality by multivariate logistic regression analysis. Results A total of 170 patients with SCAP were included in the study. The age of SCAP was 20-93(65.75±15.23) years old, and the proportion of SCAP in the elderly was 58.82%(100/170). In-hospital mortality of non-elderly SCAP was 24.3%(17/70), and the in-hospital mortality of elderly SCAP was 28%(28/100). Compared with non-elderly group, patients in elderly group had higher severity score and more complications on admission, but the symptoms of fever and respiratory rate at admission were less obvious. In multivariable logistic regression analysis, the factors significantly associated with in-hospital mortality of non-elderly SCAP were pneumonia severity index(PSI) score(P=0.016,OR=1.022, 95%CI1.004-1.041) and invasive mechanical ventilation(P=0.037,OR=4.543, 95%CI1.092-18.898) on admission, and the risk factors associated with in-hospital mortality in elderly SCAP were sequential organ failure assessment(SOFA) score(P=0.006,OR=1.240, 95%CI1.063-1.446) and combined with coronary artery disease on admission(P=0.037,OR=2.834, 95%CI1.066-7.534). Conclusion In-hospital mortality for SCAP is high. PSI score and invasive mechanical ventilation are risk factors for in-hospital mortality of non-elderly patients with SCAP, and SOFA score and combined with coronary artery disease on admission are risk factors for in-hospital mortality of elderly patients with SCAP.

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Kidney stones are common and frequently occurring diseases in urology. In recent years, the incidence of urinary stones has increased year by year, affecting people′s quality of life and aggravating the social and economic burden. The occurrence and development of kidney stones are closely related to metabolic factors. This review mainly conducts a systematic overview of the research on the relationship between electrolyte metabolism abnormalities, dyslipidemias and other metabolic disorder diseases and kidney stones in recent years.By exploring the relationship between various metabolic disorder diseases and kidney stones, this review provides reference for exploring the occurrence and development of kidney stones from the perspective of metabolism.

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Alzheimer′s disease(AD) is a chronic and progressive neurodegenerative disorder. Since there are currently no effective treatment options, early diagnosis is crucial for slowing the progression of the disease. The retina and the brain share a common embryological origin and exhibit similarities in both anatomical structure and vascular system. In the early stages of AD, patients may develop visual symptoms due to changes in the visual cortex circuitry and associated retinal alterations. Therefore, in recent years, there has been growing interest in exploring the role of the retinal vascular network in the pathogenesis, early diagnosis, and precision intervention of neurodegenerative diseases. This review overviews the changes in the visual cortex circuitry and retinal vascular network in AD patients, their underlying mechanisms, the relationship between neural circuits and visual disorders, and introduces novel diagnostic methods in retinal research.

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Cholestasis can be seen in many acute and chronic liver diseases. If not intervened in time, persistent cholestasis can further progress to liver fibrosis, liver cirrhosis, liver failure, and even death. The pathogenesis of cholestasis is complex, and there is currently a lack of effective therapeutic drugs. Bile acids(BAs), the main component of bile, are synthesized from cholesterol in the liver as primary bile acids. After entering the intestine through the enterohepatic circulation, they are reshaped into secondary bile acids by gut microbiota. BAs can directly or indirectly affect the composition and function of gut microbiota, which in turn can regulate the synthesis and metabolism of BAs. The interaction between BAs and gut microbiota plays an important role in the pathogenesis and treatment of cholestasis. This review elucidates the bidirectional regulatory mechanisms between BAs and gut microbiota and their roles in the pathogenesis and treatment of cholestasis, aiming to provide insights and references for basic research and clinical practice related to cholestasis-associated diseases.

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Among women of reproductive age, pelvic inflammatory disease is a prevalent gynecological condition. In its early stages, it may be mild or asymptomatic. If treatment is delayed, there is a high likelihood of recurrence, which can have a major impact on women′s reproductive health and raise the financial, emotional, and physical load on the family. Significant advancements in the treatment of chronic pelvic inflammatory disease have been made possible by the thorough investigation of this condition. In an effort to advance the health management of this illness and offer fresh concepts for clinical treatment, this review discusses a number of facets of both Chinese and Western medicine.