Objective To investigate the impact of Toxoplasma gondii type I, II and III rhoptry protein 16 (ROP16) on programmed cell death ligand 1 (PD-L1) expression in lung adenocarcinoma cells, and to examine the effects of T. gondii type I ROP16 protein on the relative PD-L1 expression, the relative PD-L1 distribution on the cell membrane surface, and the binding of programmed cell death 1 (PD-1) to PD-L1 in lung adenocarcinoma cells. Methods Lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins were generated, and transfected into the human lung adenocarcinoma A549 cell line. A549 cells were used as a blank control group, and A549 cells transfected with an empty lentiviral expression vector were used as a negative control group, while A549 cells transfected with lentiviral vectors overexpressing T. gondii type I, II and III ROP16 proteins served as experimental groups. Stably transfected cells were selected with puromycin and verified using Western blotting, quantitative real-time PCR (RT-qPCR), and immunofluorescence assays. The PD-L1 expression was quantified at translational and transcriptional levels using Western blotting and RT-qPCR assays in A549 cells in the five groups, and the relative PD-L1 distribution was detected on the A549 cell membrane surface using flow cytometry. In addition, the effect of T. gondii type I ROP16 protein on the PD-1/PD-L1 binding was measured in A549 cells using enzyme-linked immunosorbent assay (ELISA). Results The relative ROP16 protein expression was 0, 0, 1.546 ± 0.091, 1.822 ± 0.047 and 2.334 ± 0.089 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 1 339.00,P < 0.001), and the relative ROP16 mRNA expression was 2.153 ± 0.949, 2.436 ± 1.614, 14.343 ± 0.020, 12.577 ± 0.285 and 15.090 ± 0.420 in the blank control group, negative control group and the T. gondii type I, II and III ROP16 protein overexpression groups, respectively (F = 483.50,P < 0.001). The ROP16 expression was higher in the T. gondii type I, II and III ROP16 protein overexpression groups than in the blank control group at both translational and transcriptional levels (allP values < 0.001). Immunofluorescence assay revealed that T. gondii type I, II and III ROP16 proteins were predominantly localized in A549 cell nuclei. Western blotting showed that the relative PD-L1 protein expression was 0.685 ± 0.109, 0.589 ± 0.114, 1.007 ± 0.117, 0.572 ± 0.151, and 0.426 ± 0.116 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 9.46,P < 0.05), and RT-qPCR assay quantified that the relative PD-L1 mRNA expression was 1.012 ± 0.190, 1.281 ± 0.465, 1.950 ± 0.175, 0.889 ± 0.251, and 0.230 ± 0.192 in the blank control group, negative control group, and the T. gondii type I, II and III ROP16 protein overexpression groups (F = 14.18,P < 0.05). The PD-L1 expression was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group at both translational and transcriptional levels (both P values < 0.05). Flow cytometry detected that the relative distributions of PD-L1 protein were (10.83 ± 0.60)%, (11.23 ± 0.20)%, and (14.61 ± 0.50)% on the A549 cell membrane surface (F = 28.31, P < 0.05), and the relative distribution of PD-L1 protein was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and negative control group (both P values < 0.001). ELISA measured significant differences in the absorbance (A) value among the T. gondii type IROP16 protein overexpression group, the blank control group and the negative control group if the concentrations of the recombinant PD-1 protein were 0.04 (F = 10.45, P < 0.05), 0.08 μg/mL (F = 11.68, P < 0.05) and 0.12 μg/mL (F = 52.68, P < 0.05), and the A value was higher in the T. gondii type IROP16 protein overexpression group than in the blank control group and the negative control group (both P values < 0.05), indicating that T. gondii type IROP16 protein promoted the PD-L1/PD-1 binding in A549 cells in a concentration-dose manner. Conclusions T. gondii type IROP16 protein overexpression may up-regulate PD-L1 expression in A549 cells at both transcriptional and translational levels and the relative PD-L1 distribution on the A549 cell membrane surface, and affect the PD-1/PD-L1 binding in a concentration-dependent manner.

Objective To explore the value of dynamic contrast-enhanced(DCE)-MRI quantitative parameters based on differential sub-sampling with Cartesian ordering(DISCO)technology for differentiating high-and low-grade breast cancer.Methods A total of 80 patients with single breast cancer confirmed by biopsy pathology were retrospectively enrolled,including 40 cases of low-grade(L group)and 40 cases of high-grade breast cancer(H group).Then quantitative parameters obtained from DISCO-DCE-MRI before treatment were compared between groups,including extravascular extracellular volume fraction(Ve),rate constant(Kep),contrast enhancement ratio(CER),maximum slope(MaxSlope)and volume transfer constant(Ktrans),and their correlations with histological grade were analyzed.Receiver operating characteristic(ROC)curves of DISCO-DCE-MRI quantitative parameters being significantly different between groups were plotted,and the area under the curves(AUC)were calculated to evaluate their efficacy for differentiating high-and low-grade breast cancer.Results Ve(0.91[0.59,0.99]),CER(2.76±0.54)and MaxSlope(0.02[0.01,0.03])in L group were all higher than those in H group(0.52[0.34,0.73],[2.31±0.74],0.01[0.01,0.02],all P＜0.05),and no significant difference of Kep nor Ktrans was found between groups(both P＞0.05).Ve,CER and MaxSlope of breast cancer were all negatively correlated with histological grade(rs=-0.43,-0.39,-0.35,all P＜0.05),while Kep andKtranshad no significant correlation with histological grade(both P＞0.05).The AUC of Ve,CER and MaxSlope for differentiating high-and low-grade breast cancer was 0.749,0.725 and 0.700,respectively.Conclusion Among DISCO-DCE-MRI quantitative parameters,Ve,CER and MaxSlope could be used for differentiating high-and low-grade breast cancer.

ObjectiveTo investigate the chemical hazards in the production line of lithium batteries, so as to provide a scientific basis for the management of occupational-health risk and to promote the healthy and sustainable development of the lithium battery industry. MethodsAn on-site survey on the process flow of the production of lithium battery was conducted in an enterprise. Volatile organic compounds (VOCs) in the occupational environment were collected by Summa canisters, carbonates and N-methyl pyrrolidone (NMP) were collected using activated carbon tubes, and airborne metals were collected using filter membranes. VOCs, carbonates and NMP were detected by gas chromatography-mass spectrometry (GC-MS), and airborne metal elements in the dust samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). ResultsNon-targeted environmental monitoring results indicated that NMP was detected in the negative /positive electrode coating, assembly and drying filling workstations, dimethyl carbonate (DMC) was detected in the assembly, drying and electrolyte injection workstations, and ethyl methyl carbonate (EMC) was detected solely in the electrolyte injection workstation. Semi-quantitative analyses of VOCs identified 136 pollutants, including acrylonitrile and halohydrocarbons. Quantitative targeted environmental monitoring results revealed the highest geometric mean (GM) concentration of EMC (31.450 mg·m^-3) was found in the assembly and drying workstations, diethyl carbonate (DEC) was detected in all workstations. While vinylene carbonate (VC) and ethylene carbonate (EC) were detected only in electrolyte injection, assembly and drying workstations. NMP was detected in all positive electrode coating samples, with a GM concentration of 5.68 mg·m^-3 (concentration range: 4.0‒ 7.4 mg·m^-³). Lithium was exclusively detected in dust samples from the liquid injection workstation (GM: 0.014 μg·m^-³). ConclusionNMP, EMC, DEC, and other chemicals are identified at the key workstations such as the positive electrode coating, electrolyte injection, assembly and drying in the lithium production line. Furthermore, semi-quantitative VOCs analyses identified 136 pollutants, demonstrating a characteristic of multicomponent chemical exposure.

Chronic obstructive pulmonary disease(COPD),as a global health challenge,brings heavy economic and psychological burdens to patients and their families.Accurately identifying the risk factors for COPD and excluding false associations are crucial for understanding its pathogenesis and formulating prevention strategies.Mendelian randomization(MR),as a supplementary method,has shown great potential in reducing the interference of confounding factors,lowering the cost of experimental research,and avoiding experimental ethical issues.This article focuses on MR and its main derivative methods,discusses their basic principles and applicable conditions,and analyzes their application effects and limitations in COPD research in combination with specific cases,enabling MR to be more widely applied in the study of influencing factors of COPD.

Chronic obstructive pulmonary disease(COPD),as a global health challenge,brings heavy economic and psychological burdens to patients and their families.Accurately identifying the risk factors for COPD and excluding false associations are crucial for understanding its pathogenesis and formulating prevention strategies.Mendelian randomization(MR),as a supplementary method,has shown great potential in reducing the interference of confounding factors,lowering the cost of experimental research,and avoiding experimental ethical issues.This article focuses on MR and its main derivative methods,discusses their basic principles and applicable conditions,and analyzes their application effects and limitations in COPD research in combination with specific cases,enabling MR to be more widely applied in the study of influencing factors of COPD.

Objective To explore the value of dynamic contrast-enhanced(DCE)-MRI quantitative parameters based on differential sub-sampling with Cartesian ordering(DISCO)technology for differentiating high-and low-grade breast cancer.Methods A total of 80 patients with single breast cancer confirmed by biopsy pathology were retrospectively enrolled,including 40 cases of low-grade(L group)and 40 cases of high-grade breast cancer(H group).Then quantitative parameters obtained from DISCO-DCE-MRI before treatment were compared between groups,including extravascular extracellular volume fraction(Ve),rate constant(Kep),contrast enhancement ratio(CER),maximum slope(MaxSlope)and volume transfer constant(Ktrans),and their correlations with histological grade were analyzed.Receiver operating characteristic(ROC)curves of DISCO-DCE-MRI quantitative parameters being significantly different between groups were plotted,and the area under the curves(AUC)were calculated to evaluate their efficacy for differentiating high-and low-grade breast cancer.Results Ve(0.91[0.59,0.99]),CER(2.76±0.54)and MaxSlope(0.02[0.01,0.03])in L group were all higher than those in H group(0.52[0.34,0.73],[2.31±0.74],0.01[0.01,0.02],all P＜0.05),and no significant difference of Kep nor Ktrans was found between groups(both P＞0.05).Ve,CER and MaxSlope of breast cancer were all negatively correlated with histological grade(rs=-0.43,-0.39,-0.35,all P＜0.05),while Kep andKtranshad no significant correlation with histological grade(both P＞0.05).The AUC of Ve,CER and MaxSlope for differentiating high-and low-grade breast cancer was 0.749,0.725 and 0.700,respectively.Conclusion Among DISCO-DCE-MRI quantitative parameters,Ve,CER and MaxSlope could be used for differentiating high-and low-grade breast cancer.

Objective:To investigate the changes of the liver volume in patients with cirrhotic portal hypertension after splenectomy combined with devascularization, and to analyze the related causes.Methods:Clinical and imaging data of 94 patients with cirrhotic portal hypertension who underwent surgical treatment at the People's Hospital of Ningxia Hui Autonomous Region between January 2014 and May 2024 were included and analyzed before and after surgery. The cohort comprised 61 males and 33 females, aged (47±12) years. The preoperative and postoperative liver volumes were compared, and the patients were divided into two groups based on the changes in postoperative liver volume: the volume increase group ( n=51) and the volume decrease group ( n=43). Clinical data were collected including liver volume, hepatic artery diameter, spontaneous portosystemic shunt (SPSS), portal vein thrombosis (PVT), and hepatic artery dilation. Results:Compared with the volume decrease group, the postoperative liver volume [(1 157±237) cm 3 vs. (977±271) cm 3] and incidence of hepatic artery dilation [92.2%(47/51) vs. 67.4%(29/43)] in the volume increase group both increased, while the incidence of newly formed SPSS [3.9%(2/51) vs. 18.6%(8/43)] and PVT [5.9%(3/51) vs. 34.9%(15/43)] after surgery both decreased, and the differences were statistically significant (all P<0.05). After splenectomy combined with devascularization, patients with decreased liver volume exhibited aggravated conditions of serrated or wavy changes at the edge of the liver and widening of liver fissures, while patients with increased liver volume had significant reduction or disappearance of serrated or wavy changes at the edge of the liver and narrowing of liver fission. Conclusion:Splenectomy combined with devascularization can increase the postoperative liver volume in patients with cirrhotic portal hypertension, and decrease in postoperative liver volume may be related to the occurrence of SPSS and PVT.

Objective:To explore the differences of the circadian rhythm gene polymorphisms between patients with major depressive disorder and those with bipolar disorder, providing a genetic basis for differential diagnosis.Methods:70 patients who were still diagnosed with major depressive disorder after 5 years and 68 patients who were still diagnosed with bipolar disorder from Zhongda Hospital affiliated to Southeast University from 2012 to 2018 were included in this study. Single nucleotide polymorphisms (SNPs) of circadian rhythm gene were selected for genetic testing. And the differences in genotype frequency, allele frequency, and haplotypes of each SNP between major depressive disorder and bipolar disorder were analyzed using UNPHASED 3.1.7.Results:The analysis of genotype frequency revealed statistically significant differences in genotype frequency of PER1rs2253820, PER1rs2735611, PER3rs12566042, PER3rs17031614, and PER3rs79372391 between the two groups ( OR(95% CI)=2.386(1.173-4.854), 2.357(1.166-4.764), 0.351(0.176-0.703), 0.389(0.196-0.773), 0.389(0.196-0.773) respectively; all P<0.05). Haplotype analysis showed that the T-C-C-T-G haplotype, in CLOCK loci (rs12505266, rs2272073, rs3817444, rs11133389 and rs12505265) was significantly different between major depressive disorder group and bipolar disorder group ( OR(95% CI)=0.108(0.010-1.185), P=0.027). Conclusion:There are significant differences in circadian rhythm gene polymorphisms between patients with major depressive disorder and bipolar disorder. Carrying the PER1rs2253820TT and PER1rs2735611GG genotypes is a risk factors for bipolar disorder.

AIM:To investigate the effects of Xiaozhong-Zhitong mixture(XZZT)on M2 polarization and Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway in mouse microglia(BV2 cells).METHODS:The BV2 cells were divided into 5 groups:blank group,model group[lipo-polysaccharide(LPS)+hypoxia],TAK-242(resatorvid,a TLR4 inhibitor)group(LPS+hypoxia+TAK-242),XZZT group(LPS+hypoxia+XZZT),and TAK-242+XZZT group(LPS+hypoxia+TAK-242+XZZT).Flow cytometry was used to detect early apoptosis and cell cycle of BV2 cells,and immunofluorescence staining was employed to detect the positive expres-sion of M1-type marker inducible nitric oxide synthase(iNOS)and M2-type marker CD206.Western blot was utilized to detect the expression of TLR4/MyD88/NF-κB signaling pathway-related proteins,including TLR4,MyD88,NF-κB p65,phosphorylated p65(p-p65),phosphorylated transforming growth factor-β-activated kinase 1(p-TAK1),and phosphory-lated IκB kinase α/β(p-IKKα/β).RT-qPCR was used to detect the mRNA expression of interleukin-1β(IL-1β),IL-10,tumor necrosis factor-α(TNF-α),TLR4,MyD88,and NF-κB p65.RESULTS:Compared with model group,the rate of early apoptosis was significantly decreased in XZZT group(P<0.01),the percentage of cells arrested in the S phase was significantly increased(P<0.01),and the protein levels of TLR4,MyD88,NF-κB p65,p-IKKα/β,p-p65,and p-TAK1 were significantly decreased(P<0.05 or P<0.01).Additionally,IL-1β,TNF-α,TLR4,MyD88 and NF-κB p65 mRNA expression levels were significantly decreased(P<0.05 or P<0.01),while IL-10 mRNA expression was significantly in-creased(P<0.05).Compared with TAK-242 group,the average percentage of iNOS positive area was significantly de-creased,while CD206 was significantly increased in TAK-242+XZZT group(P<0.01).CONCLUSION:The XZZT has the effect of inducing M2 polarization of mouse microglia,and the mechanism may be linked to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

N1-methyladenosine(m1A)RNA methylation is critical for regulating mRNA translation;however,its role in the development,progression,and immunotherapy response of head and neck squamous cell carcinoma(HNSCC)remains largely unknown.Using Tgfbr1 and Pten conditional knockout(2cKO)mice,we found the neoplastic transformation of oral mucosa was accompanied by increased m1A modification levels.Analysis of m1A-associated genes identified TRMT61A as a key m1A writer linked to cancer progression and poor prognosis.Mechanistically,TRMT61A-mediated tRNA-m1A modification promotes MYC protein synthesis,upregulating programmed death-ligand 1(PD-L1)expression.Moreover,m1A modification levels were also elevated in tumors treated with oncolytic herpes simplex virus(oHSV),contributing to reactive PD-L1 upregulation.Therapeutic m1A inhibition sustained oHSV-induced antitumor immunity and reduced tumor growth,representing a promising strategy to alleviate resistance.These findings indicate that m1A inhibition can prevent immune escape after oHSV therapy by reducing PD-L1 expression,providing a mutually reinforcing combination immunotherapy approach.