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Objective To investigate the effect of sirtuin 2(SIRT2) on the proliferation and migration of cardiac fibroblasts(CFs)in C57BL/6 mice under angiotensin II(Ang Ⅱ) stimulation. Methods The hearts were taken from 1 to 2 days C57BL/6 milk mice. After cutting and digesting, CFs were extracted by different adhesion centrifugation. After CFs attachment, the cells were cultured under control medium and Ang Ⅱ(100 nmol/L) medium and treated using OE-SIRT2 plasmid to overexpression the SIRT2 gene. RT-qPCR was used to detect mRNA expression of SIRT2 proliferating cell nuclear antigen(PCNA), periostin(POSTN)and type Ⅰ collagen procollagen A1(Col1A1), Western blot assay was used to measure the protein expression levels of SIRT2, PCNA, POSTN and Col1A1, CCK-8 assay and EdU assay were used to evaluate CFs proliferation rate, Transwell experiment was used to assess CFs migration activity. Results Compared with control group, Ang Ⅱ stimulation led to down-regulation of SIRT2 expression in CFs, increased collagen expression, and promoted CFs proliferation and migration. The expression of SIRT2 was up regulated in CFs treated with OE-SIRT2 plasmid under Ang Ⅱ stimulation, Col1A1, POSTN and PCNA expression was down regulated, and CFs proliferation and migration ability decreased. Conclusion Overexpression of SIRT2 can inhibit the proliferation and migration of CFs under Ang Ⅱ stimulation, indicating that SIRT2 may be a key regulatory point in the onset and progression of cardiac fibrosis.

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Objective To investigate the mechanism of prostaglandin E synthase 3(PTGES3)/heat shock protein 90(HSP90) in the medial prefrontal cortex regulating obesity-related cognitive dysfunction. Methods This study consisted of clinical trials and animal experiments. In part one, obese patients scheduled for bariatric surgery, and healthy adults matching gender and age were recruited at the same time to reach 10 cases in each group. The cognitive level was assessed with trail making test part A(TMT-A) and victoria stroop tests(VST). Four-dimensional data-independent acquisition(4D-DIA) was used to screen the proteome changes in peripheral blood. In part two, forty SPF healthy male C57BL/6J mice were randomly divided into four groups: normal diet group(ND group), high fat diet induced obesity group(DIO group), DIO supplemented with the control virus group(DIO+Scramble group) and DIO supplemented with the interfering virus group(DIO+shPTGES3 group). The Morris water maze test was conducted to evaluate the cognitive behavior changes of the four groups of mice. The immunofluorescence staining was performed to detect the expression of PTGES3 and HSP90 in the medial prefrontal cortex and the activation of ionized calcium binding adapter molecule 1(IBA1)-labeled microglia. Results In the case-control study, the cognitive function of obese patients significantly decreased, and the expression of PTGES3 in peripheral blood significantly increased, while the level of PTGES3 was negatively correlated with cognitive function. In animal experiments, compared with ND group, DIO group had significantly prolonged time reaching the target platform, otherwise, the residence time in the target quadrant was shortened in the Morris water maze test. Simultaneously, there were significant increase in the expression of PTGES3 and HSP90, and the activation of IBA1 in the medial prefrontal cortex. Compared with DIO+Scramble group, mice in the DIO+shPTGES3 group spent less time reaching the target platform, and stayed longer in the target quadrant. The expression and co-localization levels of PTGES3 and HSP90 in medial prefrontal cortex significantly decreased. The activation level of microglia cells was also attenuated by PTGES3 interference. Conclusion Obesity-related cognitive dysfunction may be attributed to PTGES3/HSP90 in the medial prefrontal cortex by mediating neural inflammation.

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Objective To investigate the effects of microRNA-29a(miR-29a) in mediating the regulation of dihydroartemisinin(DHA) on the immune checkpoint molecule B7H3 in lung adenocarcinoma(LUAD). Methods The expression level and prognostic significance of B7H3 in LUAD were analyzed by public database. Small interfering RNA(siRNA) was used to knock down B7H3 in LUAD cell lines A549 and HCC827, and cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were treated with gradient concentrations of DHA(0, 5, 10, 25, 50, 100 μmol/L) for 48 h, and the half maximal inhibitory concentrate(IC50) was calculated. A549 and HCC827 cells were treated with IC50concentration of DHA for 1, 2 and 3 days, and the cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were transfected with miR-29a inhibitor. After DHA treatment, the expression level of miR-29a was detected by RT-qPCR, and the expression level of B7H3 was detected by Western blot. Results B7H3 was overexpressed in LUAD and associated with poor prognosis. After knocking down of B7H3, the proliferation ability of A549 and HCC827 cells significantly decreased(allP<0.001). DHA inhibited the proliferation of A549 and HCC827 cells in both dose-and time-dependent manners, with IC50values of 30.16 μmol/L and 7.50 μmol/L, respectively. DHA up-regulated the expression of miR-29a in A549 and HCC827 cells(P<0.001,P<0.01), and down-regulated the expression of B7H3 in both cell lines(P<0.01,P<0.001). After transfection of miR-29a inhibitor into A549 and HCC827 cells, the expression of B7H3 was up-regulated, and the down-regulation of B7H3 by DHA was partially reversed. Conclusion miR-29a mediates the molecular regulation of DHA on B7H3 in LUAD.

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Objective To establish anin vitromodel of ferroptosis induced by Erastin and RAS-selective lethal 3(RSL3) in hepatoma cells, and to provide theoretical basis for the development of novel therapeutic strategies for HCC. Methods Hepatoma cells(HCCLM3, HepG2, Hep3B, Huh7 and PLC/PRF/5) in logarithmic growth phase were treated with Erastin(0-40 μmol/L) and RSL3(0-10 μmol/L) at double concentrations respectively. After 24 h, CCK-8 method was used to detect cell viability, draw growth curve, calculate IC50, and HCC cells sensitive to inducers were selected for follow-up experiments. The effect of inducer on the state of hepatoma cells was observed under light microscope, and immunoblotting and flow cytometry were used to verify whether the ferroptotic modelin vitrowas successfully constructed. Results Huh7, Hep3B and HepG2 cells were sensitive to Erastin and RSL3, but HCCLM3 and PLC/PRF/5 were insensitive to Erastin and RSL3. When the concentration of Erastin and RSL3 reached the maximum, the survival rate was still above 65%. Huh7, Hep3B and HepG2 cells were selected for subsequent experiments. Compared with the control group, the expression of Glutathione peroxidase 4(GPX4), a ferroptotic marker, was down-regulated in a concentration-dependent manner. In Huh7, Hep3B and HepG2 cells, lipid reactive oxygen species(ROS) levels significantly increased after 24 h treatment with 10 μmol/L and 20 μmol/L Erastin, respectively; in Huh7 cells, lipid ROS levels significantly increased after 24 h treatment with 0.5 μmol/L and 1 μmol/L RSL3, respectively; in Hep3B and HepG2 cells, lipid ROS levels significantly increased after 24 h treatment with 1 μmol/L and 2 μmol/L RSL3, respectively, compared with control group. Conclusion Huh7, Hep3B and HepG2 cells are highly sensitive to Erastin and RSL3. Huh7, Hep3B and HepG2 cells treated with 10 μmol/L Erastin for 24 h are good models for simulating ferroptosis induced by Erastinin vitro, Huh7 cells treated with 0.5 μmol/L RSL3 for 24 h and Hep3B and HepG2 cells treated with 1 μmol/L RSL3 for 24 h are good models for simulating ferroptosis induced by RSL3in vitro.

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Objective To investigate the distribution of solute carrier organic anion transporter family member 1B1(SLCO1B1) and apolipoprotein E(ApoE) gene polymorphisms in the patients of Han ethnic group with cardiovascular and cerebrovascular diseases from Anhui Province, in order to provide the basis for the individualized therapy of statins in clinical practice. Method s 924 Han patients with cardiovascular and cerebrovascular diseases were selected. The SLCO1B1 and ApoE genotypes of the patients were detected by polymerase chain reaction-fluorescent probe method, and their distribution was compared among different genders and other regions in China. Result s Seven SLCO1B1 gene subtypes were detected in 924 patients, including *1a/*1b(33.01%),*1b/*1b(41.45%), *1b/*15(12.34%), *1a/*1a(7.03%), *1a/*15(5.52%), *15/*15(0.54%) and *5/*5(0.11%), without detection of the two gene subtypes of *1a/*5 and *5/*15; the normal metabolic genotype I of SLCO1B1(*1a/*1a, *1a/*1b, *1b/*1b) accounted for the highest proportion in this population(81.49%), the intermediate metabolic genotype II and the weak metabolic genotype III of SLCO1B1 accounted for 17.86% and 0.65% respectively; six ApoE gene subtypes were detected, including E3/E3(66.78%), E3/E4(19.37%), E2/E3(9.63%), E4/E4(1.84%), E2/E4(1.73%) and E2/E2(0.65%); the E3 mass genotype(E2/E4, E3/E3) accounted for the highest proportion in this population(68.51%); there was no significant difference in the distribution of SLCO1B1 and ApoE genes between different genders; there was no significant difference in the distribution of SLCO1B1 between the Han population from Anhui and the South China and Central China, but a significant difference was found between the Anhui Han population and the Southwest China(P<0.05); the distribution of ApoE in the Anhui Han population demonstrated no statistically significant variation from those in South China and Southwest China, whereas significant differences were observed in comparison with Central China(P<0.05). Conclusio n In the Han population with cardiovascular and cerebrovascular diseases in Anhui, the distributions of SLCO1B1 and ApoE gene polymorphisms show no significant gender differences but exhibit regional variations. These populations are predominantly characterized by class I normal metabolic genotype(SLCO1B1) and E3 mass genotypes(ApoE), indicating a higher tolerance to statin dosages and normal therapeutic efficacy in this cohort.

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Objective To predict and validate key targets for cisplatin(DDP) resistance in colorectal cancer(CRC) to provide more options for precision medicine in clinical treatment. Methods Differentially expressed genes(DEGs) between normal colonic mucosa and CRC were screened from the gene expression omnibus(GEO) database. Key genes were identified using the STRING database and Cytoscape software. DEGs were subjected to enrichment analysis using the gene ontology(GO) and kyoto encyclopedia of genes and genomes(KEGG) databases. Key targets were validated through RNA-seq, qRT-PCR, and Western blot. The versican(VCAN) gene overexpression vector was transfected into human ileocecal colorectal adenocarcinoma cell line HCT8, and cell viability was assessed using the CCK-8 assay. Flow cytometry was used to assess apoptosis and cell cycle distribution. qRT-PCR and Western blot were performed to detect mRNA and protein levels of the target genes. Results In this study, 118 upregulated DEGs and 146 downregulated DEGs were identified from the GEO database. DEGs were mainly enriched in extracellular matrix degradation, extracellular matrix organization, and the phosphoinositide 3-kinase(PI3K)-protein kinase B(AKT) signaling pathway. Based on protein-protein interaction network analysis, 20 hub genes were identified. By comparing the transcriptome sequencing results of the HCT8 parental strain and DDP-resistant strain, the VCAN gene was further selected. In CRC tissues, the expression level of VCAN was higher than that in normal colonic mucosa, and patients with high VCAN expression had shorter overall survival(OS) and recurrence free survival(RFS) times. Overexpression of VCAN in CRC cells promoted cell proliferation(P<0.05), increased resistance to DDP, reduced DDP-induced apoptosis(P<0.05), and G0/G1phase arrest(P<0.05); upregulation of VCAN activated the protein kinase B(AKT)-mammalian target of rapamycin(mTOR) signaling pathway. Conclusion Bioinformatics and transcriptome sequencing identified VCAN as a key target gene for DDP resistance in CRC, potentially promoting CRC progression and DDP resistance by regulating the AKT-mTOR pathway.

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Objective To investigate the effects of heat shock protein 70 ku protein 1A(HSPA1A) on H9c2 inflammation and apoptotic injury in rat cardiomyocytes under hypoxia, and to analyze its mechanism. Methods H9c2 cells were treated with normal oxygen(Nor) and hypoxia(Hyp), and the expression of HSPA1A was detected by RT-qPCR and Western blot. Normal H9c2 cells were divided into Nor group(normoxia culture), sh-HSPA1A+Nor group(cells transfected with HSPA1A shRNA, normoxia culture), Hyp group(hypoxia culture), sh-HSPA1A+Hyp group(cells transfected with HSPA1A SHRNA, hypoxia culture), the expression level of HSPA1A in H9c2 cells in each group was detected by RT-qPCR and Western blot, the morphology of H9c2 cells in each group was observed by inverted microscope, and the activity of H9c2 cells in each group was detected by CCK-8, the contents of inflammatory factors tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-1β and the activities of myocardial injury markers lactate dehydrogenase(LDH), creatine kinase(CK) in the supernatant of H9c2 cells were detected by ELISA, apoptosis rate of H9c2 cells in each group was detected by Annexin V-FITC/PI double staining, the expression levels of TNF receptor-associated factor 2(TRAF2)/nuclear factor κB(NF-κB) pathway-related proteins in H9c2 cells of each group were detected by Western blot. Results Compared with Nor group, the relative expression of HSPA1A mRNA and protein in H9c2 cells in Hyp group after hypoxia induction was significantly up-regulated(P<0.05). Compared with Nor group, the number of H9c2 cells in Hyp group significantly decreased, some cells were wrinkled and disordered, the cell activity significantly decreased(P<0.05), the contents of TNF-α, IL-6, IL-1β and the activities of LDH and CK in supernatant significantly increased(P<0.05), the cell apoptosis rate significantly increased(P<0.05), the relative expressions of TRAF2 protein and the phosphorylation level of p65 and inhibitor of nuclear factor κB alpha(IκBα) were significantly up-regulated(P<0.05); compared with Hyp group, the morphology of H9c2 cells in sh-HSPA1A+Hyp group was improved, the cell arrangement was more dense, the cell activity significantly increased(P<0.05), the contents of TNF-α, IL-6 and IL-1β in supernatant decreased, the activities of LDH and CK significantly decreased(P<0.05), the apoptosis rate significantly decreased(P<0.05), the relative expressions of TRAF2 protein and the phosphorylation level of p65 and IκBα were also significantly down-regulated(P<0.05). Conclusion The expression of HSPA1A in rat cardiomyocytes H9c2 is increased by hypoxia, and inhibition of HSPA1A expression can improve the hypoxia-induced inflammatory response of H9c2 cells and reduce apoptosis damage, which may be related to the regulation of TRAF2/NF-κB pathway.

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Objective To explore the differential lipid metabolites in the plasma of mice with idiopathic pulmonary fibrosis(IPF). Methods Thirty SPF C57BL/6 male mice were randomly divided into 2 groups with 15 mice in each group. The experimental groups were divided into control group and bleomycin(BLM) group. The model of idiopathic pulmonary fibrosis was induced by one-time intratracheal infusion of BLM(1 mg/kg). Hematoxylin-eosin(HE) staining was used to observe the lung histopathology. The collagen fiber deposition in lung tissue was observed by Sirius red staining. The differential lipid metabolites in plasma of IPF mice were screened and enriched by lipidomics. Results HE staining showed that the pulmonary tissue structure was disordered, alveolar septum was broken and alveolar wall was destroyed in BLM group. Sirius red staining showed a large amount of collagen fiber deposition in the lung interstitium of BLM group. The results of lipidomics analysis showed that the lipid metabolism profile of BLM group changed, 15 differential lipid metabolites were screened out, of which 11 differential lipid metabolites were up-regulated, and 4 differential lipid metabolites were down-regulated, mainly concentrated in glycerophosphoglycerophosphates, glycerophosphocholines, steroid lactones, etc. Conclusion The lipid metabolism profile of BLM group mice changes, differential lipid metabolites such as phosphoglycolate phosphatase(PGP)(18:0/18:0), PGP(i-12:0/i-24:0), PGP(i-13:0/a-25:0), and phosphatidylcholine(PC)(18:0/14:0), PC(18:3/16:0), lysophosphatidylcholine(LPC)(16:1), and LPC(18:3) may play an important role in the progression of IPF. These findings provide a new reference for further study of the molecular mechanism of IPF, and also provide a potential new target for clinical treatment.

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Objective To explore the central role of interleukin(IL)-25 in allergy of eosinophil asthma. Methods Forty 4-5 week-old C57BL/6 mice were used in this study(n=10). Mice were randomly divided into 4 groups: WT+shRNA NT group, WT+HDM group, IL-25 shRNA+IL-25 group, and IL-25 shRNA+IL-25+HDM group. According to each group′s treatment requirement, mice were treated with house dust mite and/or IL-25 by intranasal infusion. HE staining and PAS staining were used for lung tissue section staining and pathological analysis. The levels of type 2 cytokines in bronchoalveolar lavage fluid(BALF) were determined by ELISA. Flow cytometry(FCM) was used to determine the number of type 2 helper T(Th2) cells, the group 2 innate lymphoid cells(ILC2) and eosinophils in lung tissues. Mice were treated with FITC-labeled DQ-ovalbumin(DQ-OVA) by intranasal infusion to determine the antigen presentation ability of eosinophils infiltrated in the lungs. Results HE staining and PAS staining results showed that, compared with WT+shRNA NT group, a large amount of mucus and a large number of eosinophil infiltration were found in the lungs, and subcutaneous thickening of inflammatory airway were observed in the pulmonary vessels, alveolar ducts and the whole alveoli in IL-25 shRNA+IL-25+HDM group. In the IL-25 shRNA+IL-25 group, only a small amount of mucus and a small amount of eosinophil infiltration were found in the lungs, and a small amount of subcutaneous thickening of inflammatory airway were observed in pulmonary vessels, alveolar ducts and the whole alveoli. There was no significant difference in WT+HDM group. Compared with WT+shRNA NT group, the levels of IL-25, IL-4,IL-5,IL-13, IL-33 and interferon-γ(IFN-γ) in BALF significantly increased; the levels of infiltrated Th2, ILC2 and eosinophils in the lung significantly increased; the fluorescence levels of FITC-labeled DQ-OVA in eosinophils infiltrated in lungs significantly increased in IL-25 shRNA+IL-25+HDM group(P<0.05). IL-25 shRNA+IL-25 group mice also showed an increase in the above measurements, but the amplitude were limited(P<0.05). WT+HDM group had no significant difference. Conclusion IL-25 plays an essential role in the allergy and eosinophil antigen presentation processes in eosinophil asthma mice.

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Objective To investigate the role of AdipoRon, an adiponectin receptor agonist, in treatment of carbon tetrachloride(CCl4) induced liver fibrosis mice model and the mechanisms. Methods Forty mice were randomly divided into control group, model group, L-AdipoRon group and H-AdipoRon group, with 10 mice in each group. Hepatic fibrosis was induced by intraperitoneal injection of CCl4solution. The mice in L-and H-AdipoRon groups were given 100 mg/kg and 200 mg/kg AdipoRon by gavage, respectively. The activities of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) were detected by biochemical method. Liver histopathological changes and fibrosis were detected by HE staining, Masson staining and Sirius scarlet stain. The protein expression levels of Collagen I, α-smooth muscle actin(α-SMA), matrix metalloproteinase 1(MMP-1) and matrix metalloproteinase inhibitor 1(TIMP-1) in mice liver were detected by Western blot. Lipid deposition in liver were detected by oil red O staining. The percentage(%) of CD68+ iNOS+ positive M1-type macrophages in the liver were detected by immunofluorescence. The expression levels of fatty acid synthetase(Fasn), stearoyl-CoA desaturase 1(Scd1), fatty acid transporter(Cd36), peroxissome proliferator activated receptor-α(Pparα) and carnitine palmitoyl transferase 1α(Cpt1α) in mice liver tissues, as well as M1 macrophage-related genes interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) and M2 macrophage-related genes arginase 1(Arg1), Chil3 chitinase-like 3(Ym-1) were detected by RT-qPCR assay. Results Compared with model group, in low-dose AdipoRon group and high-dose AdipoRon group, serum ALT and AST activities significantly decreased(P<0.05); liver tissues structure were damaged, liver cells degeneration and inflammatory cells infiltration were improved; collagen fiber deposition was also significantly reduced; the relative expression levels of Collagen I, α-SMA and TIMP-1 proteins were significantly down-regulated(P<0.05), while the relative expression levels of MMP-1 protein were significantly up-regulated(P<0.05); the lipid droplets deposition in livers were significantly reduced. The relative Fasn, Scd1 and Cd36 mRNA expression levels in liver tissues were significantly down-regulated(P<0.05), and the relative Pparα and Cpt1α mRNA expression levels were significantly up-regulated(P<0.05); the percentage(%) of CD68+ iNOS+ positive M1-type macrophages significantly decreased(P<0.05); the relative IL-6 and TNF-α mRNA expression levels significantly decreased(P<0.05), the relative Arg1 and Ym-1 mRNA expression levels were significantly up-regulated(P<0.05). In addition, the improvement effects of high-dose AdipoRon group were better than those of low-dose AdipoRon group(P<0.05). Conclusion AdipoRon can improve the disorder of lipid metabolisms, inhibit the M1 type macrophages polarization, and improve the liver fibrosis in CCl4-induced liver fibrosis mice model.

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Objective To investigate the relationship between the risk occurrence of rectal cancer and the single nucleotide polymorphisms(SNP) of zinc finger protein 6(ZBED6), and to provide the experimental basis for early diagnosis and treatment of rectal cancer. Methods Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) technology was used to genotype the ZBED6 gene in 109 randomly selected rectal cancer patients and 110 unrelated healthy controls. To evaluate the relationship between alleles, genotypes and the risk of rectal cancer, unconditional Logistic regression analysis was used toORand 95%CI. Results The SNP rs7552670 of ZBED6 had significant correlation with the risk of rectal cancer. Compared with the population carrying the TT genotype, those carrying the TC genotype had a 2.653 fold increased risk of rectal cancer(TTvsTC:OR=2.635, 95%CI=1.501-4.690). Other SNPs had no significant correlation with the risk of rectal cancer. Conclusion There is an interaction between the polymorphisms of ZBED6(rs7552670) and rectal cancer. The population carried ZBED6 rs7552670 TC genotype had an growing risk of rectal cancer.

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Objective To explore the characteristic of obesity in adolescents with major depressive disorder and its relationship with inflammatory cytokines. Methods One hundred and forty adolescents with major depressive disorder were enrolled. According to the classification standard of body mass index(BMI) for adolescents in China, the patients were classified into underweight group, normal group, overweight group and obese group. The center for epidemiologic studies depression scale(CES-D) was used to evaluate symptoms of depression in patients, and ultrasensitive multiplex electrochemiluminescence detection technology was used to measure the levels of plasma inflammatory cytokines including interleukin(IL)-6,IL-17A,IL-1β,IL-6,IL-8 and tumour necrosis factor(TNF)-α. One-way ANOVA or Kruskal-WallisHtest and chi-square test were used for comparison between groups. Binary Logistic regression was used to analyze the influencing factors of obesity in adolescent patients with major depressive disorder. Results Among the 140 adolescent patients with major depressive disorder, wasting were 9.3%(13/140), overweight were 17.9%(25/140) and obesity were 6.4%(9/140) respectively. There were statistically significant differences in gender(χ2=8.301,P<0.05) and inflammatory cytokines IL-6(H=16.217,P<0.01), IL-8(H=10.926,P<0.05) and TNF-α(H=7.879,P<0.05) among the four groups. Analysis of covariance showed that the difference in levels of the inflammatory cytokine IL-6(F=4.486,P<0.01) remained statistically significant after controlling for age, gender and antidepressant use. The results of multiple comparisons showed that compared with the wasting group, the plasma IL-6(Z=-3.843,PBonferroni calibrate<0.01) were higher in the obese group; compared with the normal group, the obesity rate of males was higher than that of females(χ2=8.812,PBonferroni calibrate<0.01), and the level of IL-6 in the obese group(Z=-3.023,PBonferroni calibrate<0.05) was higher. Binary Logistic regression analysis showed that plasma IL-6(OR=2.500,P<0.01) and gender(OR=11.292,P<0.01) were independent influencing factors for obesity in patients with adolescent depressive disorders. Conclusion There are gender differences in obesity rates in adolescents with depressive disorders, and obesity is associated with elevated levels of inflammatory cytokines.

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Objective To investigate the molecular mechanisms and pathways of action of total flavonoids of Dracocephalum moldavica L.(TFDM) in treating vascular cognitive impairment(VCI) based on network pharmacology and in vivo animal experiments. Methods The swiss target prediction database, literature, and PubChem were used to screen the active components and action targets of TFDM. The online mendelian inheritance in man(OMIM) and GeneCards databases were utilized to screen for possible VCI targets. Venny software was used to obtain the intersection target of TFDM and VCI. The search tool for recurring instances of neighbouring genes(String) database and Cytoscape software was used to construct the PPI network. The database for annotation, visualization and integrated discovery(DAVID) database was utilized to screen for the kyoto encyclopedia of genes and genomes(KEGG) pathway and gene ontology(GO) enrichment analyses to explore the molecular mechanism and signaling pathway of TFDM for VCI. 24 rats were divided into Sham, Model, Donepezil, and TFDM groups. Except for the Sham group, the VCI model was created using modified bilateral common carotid artery ligation. After continuous gavage for 21 days, the Morris water maze test was used to evaluate the spatial learning and memory ability of rats. Hematoxy-lineosin(HE) staining was used to observe the pathological changes in the hippocampal CA1 and cortex region of the animals and immunohistochemistry detection of zonula occludens-1(ZO-1) content in the brains of the rats. Western blot was used to detect nuclear factor kappa-B p65(NF-κB p65) and tumor necrosis factor-α(TNF-α) in rat brains. Results A total of 39 active ingredients of TFDM were screened, 209 corresponding targets, 10 417 gene targets of VCI, and 193 intersecting targets. According to the results of the GO enrichment of function analysis, TFDM could improve the response of reactive oxygen species and metabolic processes of reactive oxygen species, etc. KEGG pathway enrichment analysis suggested that TFDM might regulate TNF, IL-17 signing pathway, etc. The results of animal experiments showed that TFDM improved learning and memory while reduced pathological damage in the brains of VCI rats. In addition, TFDM upregulated the positive expression of ZO-1 and downregulated the protein levels of TNF-α and NF-κB p65(P<0.05). Conclusion TFDM can improve the cognitive function of VCI through multi-components and multi-targets, and its key mechanism may be related to inhibiting TNF-α/NF-κB p65 signaling pathway,reducing neuroinflammation,and improvement of blood-brain barrier permeability.

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Objective To investigate the effect and mechanism of sex-determining region Y-box transcription factor 4(SOX4) on autophagy in small cell lung cancer(SCLC) cells. Methods Human SCLC cell line NCI-H446 was transfected with small interfering RNA(siRNA) to knockdown SOX4. RT-qPCR and Western blot were used to verify the transfection efficiency. NCI-H446 cells were divided into control group, si-SOX4 group, si-SOX4+oe-Beclin1 group and oe-Beclin1 group. Western blot analysis was performed to detect the ratio of microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ expression and the expression of Beclin1 and p62 in different groups of cells. The transcriptional regulation of Beclin1 by SOX4 was detected by dual luciferase reporter assay and chromatin immunoprecipitation PCR(ChIP-PCR) assay. CCK-8 assay was used to detect the proliferation ability of NCI-H446 cells in different groups. Flow cytometry was used to detect the apoptosis rate of NCI-H446 cells in different groups. Transwell assay was performed to determine the cell migration and invasion ability in different groups. Results Compared with the control group or the si-NC group, the relative mRNA and protein expression level of SOX4 in si-SOX4 group were down-regulated(P<0.05), and the ratio of LC3-Ⅱ/LC3-Ⅰ protein expression and the relative protein expression level of Beclin1 in si-SOX4 group decreased(P<0.05), the relative protein expression of p62 increased(P<0.05). The relative luciferase activity of Beclin1 WT in the si-SOX4 group was lower than that in the si-NC group(P<0.05); the relative enrichment of Beclin1 promoter in the Anti-SOX4 group was higher than that in the Anti-Ig G group( P<0. 05). Compared with control group,cell proliferation activity decreased,cell apoptosis rate increased,migration number and invasion number decreased in si-SOX4 group( P<0. 05). Compared with the si-SOX4 group,the ratio of LC3-Ⅱ/LC3-Ⅰ protein expression and the relative protein expression of Beclin1 increased in si-SOX4 + oe-Beclin1 group,while the relative protein expression of p62 decreased,cell proliferation activity increased,apoptosis rate decreased,migration number and invasion number increased( P<0. 05). Conclusion Down-regulation of SOX4 can inhibit autophagy,decrease proliferation activity of NCI-H446 cells,and inhibit cell migration and invasion by inhibiting Beclin1 expression.

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Objective To investigate the role of enhancer of zeste homolog 2(EZH2)-trimethylated lysine 27 of histone H3(H3K27me3) axis in the dedifferentiation of thyroid cancer and its clinical value as a potential target for the treatment of anaplastic thyroid cancer(ATC). Methods Immunohistochemical SP method was used to detect the expression of EZH2, H3K27me3, paired box gene 8(PAX8), thyroglobulin(TG) and thyroid transcription factor 1(TTF1) in ATC and papillary thyroid carcinoma(PTC) and their adjacent tissues. The relationship between EZH2 and thyroid differentiation markers(PAX8, TTF1, TG) was further analyzed by gene expression omnibus(GEO) database. ATC cell lines 8305C and BHT-101 were culturedin vitro. Real-time reverse transcription PCR(RT-qPCR) was used to detect the expression of thyroid differentiation markers(TTF1, PAX8) mRNA in ATC cell lines treated with EZH2 inhibitor(GSK126), and evaluate the potential therapeutic effect of GSK126in vitro. The effects of GSK126 and BRAF inhibitor vemurafenib on the proliferation of ATC cell lines were observed by cell proliferation assay. Results The expression of EZH2 in ATC tissues was significantly higher than that in papillary thyroid carcinoma and adjacent tissues(P<0.05). The expression of H3K37me3 in ATC tissues was significantly lower than that in PTC tissues(P<0.05). EZH2 was negatively correlated with PAX8 and TG expression levels, but not with TTF1 expression level.In vitroexperiments, GSK126 could reverse the expression of thyroid differentiation markers PAX8 and TTF1 in ATC cell lines. GSK126 combined with BRAF inhibitor vemurafenib could significantly inhibit the growth of ATC cell lines. Conclusion The EZH2-H3K27me3 axis plays an important role in regulating thyroid specific markers, and the inhibition of EZH2 by small molecular compounds is a promising target for ATC treatment in the future.

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Objective To investigate the selective activation of 5-hydroxytryptamine neurons in the dorsal raphe nucleus(DRN5-HT) of rats with insomnia induced by p-chlorophenylalanine(PCPA) using chemical genetics techniques, and to explore whether it has a regulatory effect on comorbidities of insomnia and anxiety. Methods 32 male SD rats were randomly divided into control group, PCPA model group(PCPA group), chemical activation group(hM3Dq+CNO group), and chemical control group(hM3Dq+Saline group). The anxiety level of experimental rats was evaluated through open field experiments and elevated cross maze experiments.In vivoelectroencephalography(EEG) was used to record propofol induced sleep and cortical EEG changes, and immunofluorescence staining was used to observe changes in c-Fos positive protein expression in 5-HT neurons. Results The behavioral results showed that compared with the chemical control group, the anxiety level of the chemical activation group was significantly lower(P<0.05), and sleep duration was significantly prolonged(P<0.05). The expression level of c-Fos positive protein in DRN5-HT neurons in the chemical activation group was higher(P<0.05). The EEG results showed that the percentage of δ-band power between the chemical activation group and the chemical control group was higher(P<0.05), the power percentage in the α-band was relatively low(P<0.05). Conclusion Chemical activation of 5-HT neurons in the DRN region can improve anxiety levels and increase sleep duration in PCPA induced insomnia rats.

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Objective To explore the molecular characteristics of four CAMP negativeStreptococcus agalactiae(S.agalactiae) in whole genome sequencing. Methods The identification of suspicious bacterial strains was conducted using matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS). For the strains confirmed asS.agalactiaethrough identification, further CAMP experiments were conducted. For CAMP negative strains, whole genome sequencing was performed using MGI DNBSEQ-T7 and MinION Flow Cell sequencing platforms. Subsequently, multi-locus sequence typing(MLST), virulence genes and resistance genes of the strains were compared and analyzed using various databases. Phoenix M50 fully automatic drug sensitivity analyzer was employed to determine the sensitivity of the bacterial strains to commonly used antibiotics. Results Four CAMP-negativeS.agalactiaestrains were included. Whole-genome sequencing analysis revealed that all four CAMP-negativeS.agalactiaestrains belonged to the ST862 type. These strains harbored 22 virulence genes associated with capsular polysaccharides, β-hemolysin, and hyaluronidase, as well as seven resistance genes linked to macrolides, lincosamides, polypeptides, and aminoglycosides. Antimicrobial susceptibility testing revealed that CAMP-negativeS.agalactiaewas susceptible to penicillin G, cefepime, cefotaxime, and vancomycin. However, three strains exhibited resistance to erythromycin, and one strain demonstrated resistance to clindamycin. Conclusion Four CAMP negativeS.agalactiaeof the ST862 type possess multiple virulence and drug resistance genes, showing high resistance to erythromycin, warranting clinical attention.

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Objective To investigate the value of decreased carbohydrate antigen 19-9(CA19-9) kinetics in predicting short-term outcomes and determining prognosis among advanced biliary or pancreatic cancer patients receiving first-or second-line therapy in the real world. Methods Eighty-nine patients were retrospectively collected with advanced biliary or pancreatic cancer, especially on the CA19-9 dynamics and decline rates at different time points. This study evaluated the association of CA19-9 changes with clinicopathological features, short-term response to antitumor therapy, and survival outcomes. Results The enrolled patients recorded baseline CA19-9 levels ranging from 1.20 to 65 706.40 U/ml, with a median of 303.11 U/ml. There was no statistical correlation between baseline CA19-9 levels and gender, age, body mass index, primary tumor site, hepatic metastases, pulmonary metastases, lymph node metastases, peritoneal metastases, performance status, treatment lines, and combinations of drug types. Baseline CA19-9 levels were not associated with systemic immunoinflammatory index, prognostic nutritional index, and total bilirubin. A 25% or 50% decrease in CA19-9 after 2-3 therapy courses indicated short-term efficacy in reaching tumor objective remission or disease control. Both combinations of multiple drug types and a 25% decline in CA19-9 after one course of treatment were independent prognostic factors that affected the longer progression-free survival of patients receiving first or second line of treatment. Conclusion Decreased CA19-9 kinetics has specific values in predicting the efficacy and prognosis of advanced biliary or pancreatic cancer.

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Objective To explore the polymorphism of endothelial nitric oxide synthase(eNOS) gene in umbilical cord blood of preterm infants and its relationship with major complications in preterm infants. Methods A total of 254 preterm infants(<37 weeks) who were hospitalized were selected as the study subjects. Umbilical cord blood was collected at delivery to determine the genotypes and alleles of eNOS gene at three loci: rs61722009, rs2070744,and rs1799983. Clinical data of the preterm infants were recorded, and the relationship between eNOS gene polymorphism and major complications in preterm infants was analyzed. Results(1) The TC+CC genotype at locus rs2070744 was an independent risk factor for bronchopulmonary dysplasia(BPD) in preterm infants, with an OR(95%CI) of 1.266(1.017-1.577).(2) The GT+TT genotype at locus rs1799983 was an independent risk factor for retinopathy prematurity(ROP), with an OR(95%CI) of 1.184(1.008-1.391).(3) The AB+AA genotype at locus rs61722009 was also an independent risk factor for ROP,with an OR(95%CI) of 1.335(1.033-1. 726).(4) There was no significant relationship between gene polymorphism and the occurrence of respiratory distress syndrome( RDS) and periventricular-intraventricular hemorrhage( PIVH).Conclusion e NOS gene polymorphism is associated with the occurrence of BPD and ROP in preterm infants. The evaluation of e NOS gene polymorphism by umbilical cord blood measurement is helpful for the prevention and correct management of some serious complications.

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Objective To assess the epidemiological, drug resistance, and their relative risk and prevalence for yielding clinical diseases ofStreptococcus mitisgroup(SMG) bloodstream infections in recent years. Methods A total of 50 blood culture specimens were collected from patients with SMG bloodstream infection. These SMG isolates were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS). The susceptibility to antibiotics was tested by minimal inhibitory concentrations and Kirby-Bauer(K-B) disk diffusion methods. The data were comprehensively analyzed by statistical software combined with clinical data. Results Five strains were identified in SMG bloodstream infection by mass spectrometry, namelyStreptococcus oralis/mitis(S.oralis/mitis),Streptococcus pneumonia(S.pneumonia),Streptococcus gordonii(S.gordonii),Streptococcus sanguinis(S.sanguinis), andStreptococcus parasanguinis. These SMG showed high resistance to erythromycin and clindamycin, but low resistance to penicillin, ampicillin and ceftriaxone. Reduced hemoglobin and albumin, elevated C-reactive protein and procalcitonin were the common hematological changes in patients with SMG bloodstream infections. In SMG bacteremia,S.gordonii,S.sanguinisandS.orals/mitiswere the leading group causing infective endocarditis. Patients with myocardial disease factor were more likely to cause infective endocarditis byS.gordoniiandS.sanguinis, compared withS.orals/mitis.S.oralis/mitisbacteremia more occurred in patients with renal transplants progressing to pulmonary infection. Conclusion In this area, β-lactam antibiotics are the best choice for treating SMG. SMG species with closely related phylogenetically show different prevalence and risk of clinical disease in bloodstream infection patients. Early prevention and diagnosis of bacteremia caused by SMG are necessary for clinical diagnosis, treatment and effective control of infectious diseases progression.

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Objective To explore the correlation between uteroglobulin-related protein 1(UGRP1) and primary hypothyroidism. Methods Ninety-six patients with primary hypothyroidism were selected, including 66 patients with positive thyroid peroxidase antibodies(TPOAb) or anti-thyroglobulin antibodies(ATG) as the antibody-positive group, 30 patients with negative thyroid autoantibodies as the antibody-negative group, and 96 healthy people as the control group. The general clinical data, thyroid-related indicators and serum UGRP1 levels were compared among these three groups. Human thyroid normal cells(NTHY-ORI 3-1) were transfected with plasmids in vitro, thus establishing the control group as well as the UGRP1 group. Enzyme linked immunosorbent assay(ELISA) was used to detect and compare the T4 level in the cell culture supernatant. Results The differences in thyroid stimulating hormone(TSH), TPOAb and ATG among the three groups were statistically significant(P<0.05). The serum UGRP1 levels in the antibody-positive(303.97±156.00) pg/ml and antibody-negative groups(352.13±188. 37) pg/ml were higher than those in the control group( 237. 54 ± 137. 20) pg/ml,and the differences between the groups were statistically significant( P = 0. 005). Meanwhile,there was no statistically significant difference between the antibody-positive and antibody-negative groups. Multi-factor Logistic regression analysis showed that UGRP1 was the risk factor for the occurrence of primary hypothyroidism( OR = 1. 004,95% CI: 1. 001-1. 007,P =0. 007). The difference between the control group and UGRP1 group in T4 concentration secreted by human thyroid normal cells was not statistically significant. Conclusion Serum UGRP1 levels increase in patients with primary hypothyroidism,and the high expression of UGRP1 may have no direct relation to the function of thyroid cells secreting T4.

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Objective To evaluate the clinical value of combining low-dose energy spectrum CT with virtual un-enhanced(VUE) scanning and deep-learning image reconstruction(DLIR) in aortic CT angiography(CTA). Methods In a prospective study, 94 patients scheduled for aortic CTA were randomized into two groups: a low-dose energy spectrum group and a standard 100 kVp enhancement group, with 47 patients in each. All patients initially underwent a true un-enhanced(TUE) scan at 120 kVp using adaptive statistical iterative reconstruction-V(ASIR-V) at 40% for image reconstruction. The low-dose group received enhanced scans using gemstone spectral imaging(GSI) mode with DLIR-H, producing 60 keV virtual monoenergetic images(VMIs) and VUE images. The standard group was scanned at 100 kVp, with images reconstructed using ASIR-V at 50%. Parameters were measured including CT values, noise(SD), signal-to-noise ratio(SNR), and contrast-to-noise ratio(CNR) for key vascular and muscular areas, alongside the effective radiation dose(ED). Two radiologists evaluated the image quality using a 5-point scale. Results The low-dose group exhibited significantly higher SNR and CNR values in the ascending aorta, descending aorta, abdominal aorta, and common iliac artery compared to the standard group(P<0.05), with comparable subjective quality scores. The VUE images also demonstrated superior SNR values in the abdominal aorta, common iliac artery, and psoas major muscle, and CNR value in the ascending aorta compared to TUE images, with similar subjective quality. Importantly, the ED in the low-dose group was about 40% lower than that of the standard group. Conclusion Low-dose energy spectrum CT with DLIR in aortic CTA can significantly enhance SNR and CNR, while approximating the image quality of traditional TUE scans, thereby substantially reducing radiation exposure.

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Objective To explore the predictive value of depression severity plasma thyroid-stimulating hormone(TSH) and brain-derived neurotrophic factor(BDNF) levels for agitated symptoms in patients with adolescent depressive disorder(MDD). Methods Ninety-one patients with adolescent depressive disorder were enrolled, and the degree of agitation was assessed according to the modified outward aggressive behavior scale(MOAS); 24-item hamilton depression scale(HAMD24) was used to determine the severity of depression; chemiluminescence immunoassay(CLIA) was used to determine the plasma thyroid-stimulating hormone(TSH) level; and electrochemiluminescence immunoassay(ECL) was used to determine the plasma BDNF. SPSS 26.0 was used for statistical analysis of the data, Spearman correlation analysis was used to analyze the relationship between HAMD24and plasma TSH and BDNF levels and the degree of agitation, multiple linear regression analysis was used to analyze the factors influencing the degree of agitation in adolescents with MDD, and binary Logistic regression analysis and subjects′ work characteristic curves(ROC) were used to establish predictive models. Results The degree of agitation in adolescent MDD patients was positively correlated with HAMD24total score(P<0.001); both HAMD24total score and plasma BDNF level were identified as risk factors for agitation severity(bothP<0.05); both HAMD24total score and plasma BDNF levels were risk factors for the degree of agitation(allP<0.05); HAMD24total score, plasma TSH, BDNF levels were all risk factors for concomitant agitation symptoms in adolescent MDD patients; ROC curve analysis showed that the three combined prediction models(AUC=0.889,P<0.001) had a higher predictive value than the single prediction model(P<0.01). Conclusion Concomitant agitation symptoms in adolescents with MDD are strongly associated with HAMD24total score and plasma TSH and BDNF levels, and the three combined models have good predictive power.

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Objective To investigate associations of suicidal ideation with clinical features, inflammatory markers, and thyroid hormones in patients with schizophrenia. Methods The subjects of this study were 203 schizophrenic patients, grouped on the basis of suicidal ideation. Clinical characteristics were assessed using multiple scales. Neutrophil to lymphocyte ratio(NLR), platelet to lymphocyte ratio(PLR), monocyte to lymphocyte ratio(MLR) and thyroid hormones were also detected. SPSS 23.0 was used for statistical analyses. Results The prevalence of suicidal ideation was 21.7% in patients with schizophrenia. Logistic stepwise regression analyses showed that Calgary depression scale(CDSS) total score(OR=1.490, 95%CI=1.287-1.724,P<0.001), insomnia severity index scale(ISI) total score(OR=1.096, 95%CI=1.011-1.187,P=0.025), modified overt aggression scale(MOAS) total score(OR=1.111, 95%CI=1.016-1.215,P=0.021), MLR(Ln)(OR=15.123, 95%CI=3.868-59.125,P<0.001), and triiodothyronine(T3)(OR=0.037, 95%CI=0.003-0.388,P=0.006) were the independent influences of suicidal ideation. Additionally, receiver operating characteristic(ROC) curve analyses revealed that the five-item combination of CDSS total score, ISI total score, MOAS total score, MLR(Ln), and T3(AUC=0.908, 95%CI=0.867-0.949,P<0.001) had better ability to identify suicidal ideation. Conclusion The risk of suicidal ideation is relatively high in patients with schizophrenia, and there may be stronger associations between suicidal ideation and depression, insomnia, aggression, MLR, and T3.

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Non-small cell lung cancer(NSCLC), the most prevalent type of lung cancer, has a poor prognosis in patients with advanced disease. In recent years, immune checkpoint inhibitors(ICIs) have demonstrated promising efficacy in this disease, while bringing a unique set of immune-related adverse events(irAEs). This article comprehensively explores the multi-systemic irAEs of programmed death protein-1(PD-1)/programmed death protein ligand-1(PD-L1) inhibitors in the treatment of NSCLC, including but not limited to dermatotoxicity, endocrine toxicity,hepatic toxicity and gastrointestinal toxicity. The occurrence of these adverse reactions not only poses a challenge for clinical treatment,but also correlates with treatment efficacy. In addition,the paper discusses biomarkers for predicting the risk of ir AEs,such as gut microbiota,blood biomarkers,etc.,with the aim of providing a potential risk assessment tool for the clinic.

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Alzheimer′s disease(AD) is a common neurodegenerative disease. It has been found that AD is related to various pathogenic factors such as genetics, cardiovascular and cerebrovascular disease, and excessive phosphorylation of tau protein. However, no definitive conclusions on its pathogenesis have been reached. In this paper, the research progress on the pathogenesis of AD inC.elegansmodel and the therapeutic effects of traditional Chinese medicine extracts on AD are reviewed, providing a basis for further research on the alleviating effects of Chinese medicine extracts on AD.

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Asthma is a well-characterized heterogeneous disease marked by airway remodeling and chronic airway inflammation. Clinically, the treatment of asthma primarily relies on hormonal drugs. However, the long-term use of these medications can lead to significant side effects. Mitophagy is a biological process that selectively transports damaged mitochondria to lysosomes for degradation. Recent research has revealed the crosstalk between mitophagy and asthma. Accordingly, taking mitophagy as an entry point, summarizing the key molecular mechanisms and regulators of mitophagy in asthma will facilitate the development of novel intervention targets and strategies for asthmatic treatment.