Objective: Previous studies have discovered a correlation between cigarette smoking and cortical thickness and surface area, but the causal relationship remains unclear. The objective of this investigation is to scrutinize the causal association between them. Methods: To derive summary statistics from a genome-wide association study (GWAS) on cortical thickness, surface area, and four smoking behaviors: 1) age of initiation of regular smoking (AgeSmk); 2) smoking initiation (SmkInit); 3) smoking cessation (SmkCes); 4) cigarettes per day (CigDay). Linkage disequilibrium score regression (LDSC) was employed to examine genetic association analysis. Furthermore, for traits with significant genetic associations, Mendelian randomization (MR) analyses were conducted. Results: The LDSC analysis revealed nominal genetic correlations between AgeSmk and right precentral surface area, left caudal anterior cingulate surface area, left cuneus surface area, left inferior parietal surface area, and right caudal anterior cingulate thickness, as well as between CigDay and left caudal middle frontal surface area, between SmkCes and left entorhinal thickness, and between SmkInit and left rostral anterior cingulate surface area, right rostral anterior cingulate thickness, and right superior frontal thickness (rg=-0.36–0.29, p<0.05). MR analysis showed a unidirectional causal association between left caudal middle frontal surface area and CigDay (βIVW=0.056, pBonferroni=2×10-4). Conclusion Left caudal middle frontal surface area has the potential to serve as a significant predictor of smoking behavior.

Aim To investigate the regulatory effects and underlying mechanisms of 4-methoxybenzyl alcohol(4-MBA),an active ingredient of Gastrodia elata,on hepatic cholesterol metabolism.Methods Acute hy-perlipidemia mouse models were established via egg yolk emulsion induction,and hyperlipidemia rat models were constructed using a high-fat diet.Serum and he-patic total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),and high-den-sity lipoprotein cholesterol(HDL-C)levels were quan-tified via enzymatic assays.Hepatic histopathological changes were evaluated through hematoxylin-eosin(HE)and Oil Red O staining.Interactions between 4-MBA and key cholesterol metabolism targets were sim-ulated using molecular docking.mRNA and protein ex-pression levels of LDL receptor(LDLR),proprotein convertase subtilisin/kexin type 9(PCSK9),liver X receptor α(LXRα),peroxisome proliferator-activated receptor γ(PPARγ),ATP-binding cassette transporter G1(ABCG1),and cholesterol 7α-hydroxylase(CYP7A1)were assessed using quantitative polymer-ase chain reaction(qPCR)and immunohistochemis-try.Results In acute hyperlipidemic mice,4-MBA administration significantly reduced serum TG and LDL-C levels while elevating HDL-C(P＜0.05).Hy-perlipidemic rats exhibited decreased serum TG and LDL-C,increased HDL-C(P＜0.01),reduced hepatic LDL-C(P＜0.01),and elevated hepatic HDL-C(P＜0.01).Although TC levels showed a downward trend,the difference lacked statistical significance.He-patic lipid accumulation and steatosis were alleviated.Upregulated mRNA and protein expression of LDLR,PPARγ,LXRα,and ABCG1(P＜0.01),alongside downregulated PCSK9(P＜0.05),were observed.Conclusion 4-MBA modulates cholesterol metabolism primarily via the LDLR/PCSK9 pathway to enhance cholesterol uptake and the PPARγ-LXRα-CYP7A1/ABCA1 axis to promote cholesterol utilization and ef-flux.

AIM To investigate the renal protective effects of Qizhi Tongluo Formula on a mouse model of diabetic nephropathy.METHODS The male db/db mice were randomly divided into the model group,the dapagliflozin group(0.76 mg/kg)and the low,medium and high dose Qizhi Tongluo Formula groups(7.83,15.65 and 31.3 g/kg),with 6 mice in each group,in contrast to the 6 db/m mice of the control group.When the mice of the control group and the model group were given distilled water by gavage,those of the other administration groups were dosed with the corresponding drug by gavage once daily for 8 weeks.After the drug administration,the mice had their levels of FBG,BUN,Scr and 24 h-UTP detected;their renal pathological changes observed by transmission electron microscopy(TEM)and HE staining;their levels of serum Nrf2,HO-1,Keap1 and renal oxidative stress assessed by ELISA;their renal Nrf2 protein expression observed by immunofluorescence(IF);their renal protein expressions of Nrf2,HO-1 and Keap1 detected by Western blot;and their renal Nrf2,HO-1,and Keap1 mRNA expressions detected by RT-qPCR.RESULTS Compared with the control group,the model group displayed increased levels of 24 h-UTP,Scr,FBG and renal MDA(P＜0.01);decreased renal activities of SOD,CAT and GSH-Px(P＜0.01);mild glomerular mesangial hyperplasia,vacuolated renal tubular epithelial cells,widely fused podocyte foot processes,disappearance of tear film,decreased secretion levels of serum Nrf2 and HO-1 and renal protein and mRNA expressions of Nrf2 and HO-1(P＜0.05,P＜0.01);and decreased secretion levels of serum Keap1 and renal Keap1 protein and mRNA expressions(P＜0.01).Compared with the model group,the high-dose Qizhi Tongluo Formula group demonstrated decreased levels of 24 h-UTP,Scr,FBG and renal MDA(P＜0.01);increased renal activities of SOD,CAT and GSH-Px(P＜0.01);alleviated renal pathological damage,increased secretion levels of serum Nrf2 and HO-1 and renal protein and mRNA expressions of Nrf2 and HO-1(P＜0.01);and increased level of serum Keap1 secretion and renal Keap1 protein and mRNA expressions(P＜0.01).CONCLUSION Qizhi Tongluo Formula can inhibit oxidative stress and alleviate kidney damage in db/db mice by activating Nrf2/Keap1/ARE signaling pathway.

Objective To explore the effects of triptolide(TPL)on the proliferation,migration and invasion of gastric cancer cells and its mechanism.Methods Human gastric cancer cell line MKN45 was cultured in vitro and treated with different concentrations of TPL for 48 hours.The cell proliferation inhibition rate was detected by CCK-8 method and the optimal concentration was selected for subsequent experiments.qRT-PCR was used to detect the expression of miR-29b and KDM2A mRNA in cells treated with different concentrations of TPL.MKN45 cells at logarithmic growth phase were randomly divided into the control group(without any treatment),the TPL group(treated with 200 μg/mL TPL),the inhibitor-NC+TPL group(transfected with inhibitor-NC and then treated with 200 μg/mL TPL),and the miR-29b inhibitor+TPL group(transfected with miR-29b inhibitor and then treated with 200 μg/mL TPL).qRT-PCR was used to detect the expression of miR-29b and KDM2A mRNA in each group of cells,and Western blot was used to detect the expression of KDM2A protein.The clone formation ability of each group of cells was detected by plate clone formation assay,and the migration and invasion abilities of each group of cells were detected by Transwell assay.Results TPL at concentrations of 25 μg/mL,50 μg/mL,100 μg/mL,and 200 μg/mL could significantly inhibit the proliferation of MKN45 cells(P<0.05),up-regulate the expression of miR-29b in cells(P<0.05),and down-regulate the expression of KDM2A mRNA(P<0.05).The effect was most obvious at the concentration of 200 μg/mL,so 200 μg/mL TPL was selected for the subsequent experiments.Compared with the control group,the expression of miR-29b in the TPL group increased(P<0.05),the expression of KDM2A mRNA and protein decreased(P<0.05),and the numbers of clone formation,migration and invasion cells reduced(P<0.05).Compared with the inhibitor-NC+TPL group,the expression of miR-29b in the miR-29b inhibitor+TPL group decreased(P<0.05),the expression of KDM2A mRNA and protein increased(P<0.05),and the numbers of clone formation,migration and invasion cells increased(P<0.05).Conclusion TPL can inhibit the proliferation,migration and invasion of gastric cancer cells,and its mechanism is related to the regulation of the miR-29b/KDM2A signaling pathway.

Cholera toxin B subunit(CTB)can enhance antigen presentation and promote T cell pro-liferation,B cell differentiation and B cell isotype conversion.Moreover,woodchuck hepatitis virus post transcriptional regulatory element(WPRE)can enhance gene expression efficiency by optimi-zing RNA polyadenylation,denuclearization and/or translation.In order to construct porcine epi-demic diarrhea virus like particles(VLPs)chimerized with CTB and WPRE and evaluate their im-munogenicity,the G Ⅱ type PEDV S gene,combined with the elements promoting the protein ex-pression and enhancing immune effects,was synthesized by the company and cloned into pET32a(+).Af-ter double enzyme digestion and gel recovery,the gene named as TSCW was cloned into pFastBacl to construct the recombinant plasmid pFastBac-TSCW.pFastBac-TSCW was further transformed into DH10Bac competent cells to obtain recombinant bacmid Bacmid-TSCW.Subsequently,the Bacmid-TSCW was transfected into sf9 cells to obtain recombinant baculovirus BV-TSCW.After co-infection of BV-TSCW and BV-M into sf9 cells,viral like particles VLP-TSCW was obtained and used to immunize mice to evaluate its immunogenicity.The results showed that the recombi-nant plasmid pFastBac-TSCW and bacmid Bacmid-TSCW were successfully constructed.After transfection of sf9 cells with recombinant baculovirus,significant cytopathic effects were observed.PCR and Western blot results showed that the recombinant baculoviruses existed stably in sf9 cells and the target proteins was also expressed stably.In addition,the electron microscopy results showed that BV-TSCW and BV-M successfully assembled into viral like particles VLP-TSCW.Furthermore,ELISA results indicated that VLP-TSCW induced high level specific antibodies.The above results laid the foundation for further optimization,design and development of PEDV VLPs subunit vaccines.

AIM:To investigate the therapeutic effects and potential mechanism of pachymic acid(PA)on li-popolysaccharide(LPS)-induced acute kidney injury(AKI)in mice.METHODS:(1)Genes related to AKI were screened using the DAVID database.Core genes were identified by intersecting related genes and analyzed using Cyto-scape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed through the DAVID database for the cross-targets.Molecular docking and activity assays were conducted on the primary core targets.(2)A total of 100 C57BL/6J mice were randomly divided into five groups:normal control(NC),model(LPS),solvent control(LPS+DMSO),and treatment groups(LPS+PA-10 and LPS+PA-20),with 20 mice in each group.The LPS-AKI model was established by intraperitoneal injection of 18 mg/kg LPS.The treatment groups received 10 mg/kg and 20 mg/kg PA,respectively,and the solvent control group was administered an equivalent dose of DMSO.Mice were euthanized 24 h after injection.Serum was collected for biochemical analysis,and Western blot was used to detect neutro-phil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),caspase-3,cleaved caspase-3,interleu-kin-1β(IL-1β),and monocyte chemoattractant protein-1(MCP-1)protein expression.RT-qPCR was employed to detect inflammatory factor mRNA levels.Molecular docking was used to simulate the optimal binding site of PA to caspase-3.En-zyme activity assays were performed to assess caspase protein activity,and renal lesions were observed via hematoxylin and eosin(HE)staining.Apoptosis was detected by TUNEL staining.RESULTS:(1)Thirty-one potential targets of PA against AKI were identified through network pharmacology.GO and KEGG enrichment analyses indicated that these tar-gets were primarily involved in immune response,inflammatory processes,apoptosis and survival,angiogenesis and hemo-dynamics,oxidative stress,and endoplasmic reticulum stress.Key targets included CASP3(caspase-3),PTGS2,BCL2,CCL2,and CYP219.(2)PA treatment improved renal function and reduced tubular epithelial injury.It significantly de-creased NGAL,KIM-1,and cleaved caspase-3 protein levels,as well as inflammatory factors TNF-α,IL-1β,and MCP-1 mRNA and protein expression.PA also reduced apoptosis of renal tubular epithelial cells.Enzyme activity assays and mo-lecular docking revealed that PA exerted its anti-apoptotic effect by directly binding to caspase-3,thereby inhibiting its ac-tivation by caspase-8.CONCLUSION:PA demonstrated a therapeutic effect in LPS-AKI,potentially through the inhibi-tion of inflammatory factor synthesis and release,as well as the inhibition of caspase-3 activation by caspase-8,reducing apoptosis in renal tubular epithelial cells.

Objective:To explore the predictive value of ultrasonic shear wave elastography (SWE) parameters and pathological characteristics on the status of human epidermal growth factor receptor 2 (HER2), which is difficult to be determined by immunohistochemistry (IHC) in breast cancer, and to construct a nomogram model.Methods:A retrospective case-control study was conducted. One hundred and fifteen cases of breast cancer diagnosed and treated in Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University from September 2018 to April 2022 were selected, and their HER2 was evaluated as IHC 2+; the HER2 expression status was determined by fluorescence in situ hybridization (FISH) detection, including 23 HER2 positive cases and 92 HER2 negative cases. The ultrasound SWE parameters [including maximum shear wave velocity (V max), mean shear wave velocity (V mean), median shear wave velocity (V median), minimum shear wave velocity (V min)] and clinicopathological characteristics between HER2 positive and negative groups were compared. The variables with statistically significant differences ( P < 0.05) between groups were included in a multivariate logistic regression model, the independent risk factors for HER2 positivity were screened, and a nomogram model was constructed based on these independent risk factors. With the FISH test results as the gold standard, the efficacy of nomogram in judging HER2 positivity in breast cancer which was difficult to be identified by IHC was evaluated with the receiver operating characteristic (ROC) curve; the accuracy and clinical net benefit of the nomogram model were evaluated using calibration curve and decision curve analysis (DCA), respectively. Results:The patients were all female, aged (56±13) years, ranging from 30 to 88 years old. V max [ M ( Q1, Q3)] [8.54 (7.38, 9.47) m/s vs. 6.46 (5.07, 8.42) m/s], V mean [(5.41±0.78) m/s vs. (4.53±1.22) m/s], V median [5.06 (4.48, 5.52) m/s vs. 4.35 (3.42, 4.96) m/s], V min [3.35 (2.68, 3.88) m/s vs. 2.59 (2.11, 3.34) m/s], the proportion of patients with axillary lymph node metastasis [56.5% (13/23) vs. 22.8% (21/92)], and the Ki-67 positivity index [35% (30%, 55%) vs. 25% (15%, 35%)] in the HER2 positive group were higher than those in the HER2 negative group, and the differences were statistically significant (all P < 0.05); There was no statistically significant difference in age, lesion location, pathological type, vascular invasion, nerve invasion and long diameter, short diameter, echo, regular shape, clear boundary, posterior echo, calcification, blood flow grading, Breast Imaging Report and Data System (BI-RADS) classification detected by ultrasound between the two groups (all P > 0.05). Multivariate logistic regression analysis showed that increased ultrasound V max ( OR = 1.786, 95% CI: 1.283-2.485, P = 0.001) and axillary lymph node metastasis ( OR = 4.185, 95% CI: 1.327-13.197, P = 0.015) and elevated Ki-67 positivity index ( OR = 1.042, 95% CI: 1.014-1.071, P = 0.003) were independent risk factors for HER2 positivity. ROC curve analysis showed that the area under the curve (AUC) of HER2 positive breast cancer which was difficult to be determined by IHC was 0.816 (95% CI: 0.732-0.883), that was higher than 0.712 (95% CI: 0.620-0.794) of V max, 0.601 (95% CI: 0.504-0.692) of axillary lymph node metastasis and 0.706 (95% CI: 0.613-0.788) of Ki-67 positivity index based on the nomogram constructed by the above independent risk factors, with statistically significant differences (all P < 0.05). The calibration curve showed that the predicted probability of the nomogram model was close to the actual probability, and DCA indicated that the clinical net benefit of the model was good. Conclusions:The nomogram constructed based on ultrasonic SWE parameter V max, axillary lymph node metastasis and Ki-67 positivity index has a good predictive effect on HER2 status of breast cancer which is difficult to be determined by IHC.

Objective: Previous studies have discovered a correlation between cigarette smoking and cortical thickness and surface area, but the causal relationship remains unclear. The objective of this investigation is to scrutinize the causal association between them. Methods: To derive summary statistics from a genome-wide association study (GWAS) on cortical thickness, surface area, and four smoking behaviors: 1) age of initiation of regular smoking (AgeSmk); 2) smoking initiation (SmkInit); 3) smoking cessation (SmkCes); 4) cigarettes per day (CigDay). Linkage disequilibrium score regression (LDSC) was employed to examine genetic association analysis. Furthermore, for traits with significant genetic associations, Mendelian randomization (MR) analyses were conducted. Results: The LDSC analysis revealed nominal genetic correlations between AgeSmk and right precentral surface area, left caudal anterior cingulate surface area, left cuneus surface area, left inferior parietal surface area, and right caudal anterior cingulate thickness, as well as between CigDay and left caudal middle frontal surface area, between SmkCes and left entorhinal thickness, and between SmkInit and left rostral anterior cingulate surface area, right rostral anterior cingulate thickness, and right superior frontal thickness (rg=-0.36–0.29, p<0.05). MR analysis showed a unidirectional causal association between left caudal middle frontal surface area and CigDay (βIVW=0.056, pBonferroni=2×10-4). Conclusion Left caudal middle frontal surface area has the potential to serve as a significant predictor of smoking behavior.

Objective: Previous studies have discovered a correlation between cigarette smoking and cortical thickness and surface area, but the causal relationship remains unclear. The objective of this investigation is to scrutinize the causal association between them. Methods: To derive summary statistics from a genome-wide association study (GWAS) on cortical thickness, surface area, and four smoking behaviors: 1) age of initiation of regular smoking (AgeSmk); 2) smoking initiation (SmkInit); 3) smoking cessation (SmkCes); 4) cigarettes per day (CigDay). Linkage disequilibrium score regression (LDSC) was employed to examine genetic association analysis. Furthermore, for traits with significant genetic associations, Mendelian randomization (MR) analyses were conducted. Results: The LDSC analysis revealed nominal genetic correlations between AgeSmk and right precentral surface area, left caudal anterior cingulate surface area, left cuneus surface area, left inferior parietal surface area, and right caudal anterior cingulate thickness, as well as between CigDay and left caudal middle frontal surface area, between SmkCes and left entorhinal thickness, and between SmkInit and left rostral anterior cingulate surface area, right rostral anterior cingulate thickness, and right superior frontal thickness (rg=-0.36–0.29, p<0.05). MR analysis showed a unidirectional causal association between left caudal middle frontal surface area and CigDay (βIVW=0.056, pBonferroni=2×10-4). Conclusion Left caudal middle frontal surface area has the potential to serve as a significant predictor of smoking behavior.

Objective: Previous studies have discovered a correlation between cigarette smoking and cortical thickness and surface area, but the causal relationship remains unclear. The objective of this investigation is to scrutinize the causal association between them. Methods: To derive summary statistics from a genome-wide association study (GWAS) on cortical thickness, surface area, and four smoking behaviors: 1) age of initiation of regular smoking (AgeSmk); 2) smoking initiation (SmkInit); 3) smoking cessation (SmkCes); 4) cigarettes per day (CigDay). Linkage disequilibrium score regression (LDSC) was employed to examine genetic association analysis. Furthermore, for traits with significant genetic associations, Mendelian randomization (MR) analyses were conducted. Results: The LDSC analysis revealed nominal genetic correlations between AgeSmk and right precentral surface area, left caudal anterior cingulate surface area, left cuneus surface area, left inferior parietal surface area, and right caudal anterior cingulate thickness, as well as between CigDay and left caudal middle frontal surface area, between SmkCes and left entorhinal thickness, and between SmkInit and left rostral anterior cingulate surface area, right rostral anterior cingulate thickness, and right superior frontal thickness (rg=-0.36–0.29, p<0.05). MR analysis showed a unidirectional causal association between left caudal middle frontal surface area and CigDay (βIVW=0.056, pBonferroni=2×10-4). Conclusion Left caudal middle frontal surface area has the potential to serve as a significant predictor of smoking behavior.